Analyzing Access and Costs of Oral Medications for Overactive Bladder: Uncovering Disparities.
To report out-of-pocket costs associated with overactive bladder (OAB) medications among Medicare beneficiaries and the uninsured. We performed a cross-sectional analysis of the Centers for Medicare & Medicaid Services Prescription Drug Plan Formulary Data (Q1-2022). FDA-approved medications for OAB were identified. We calculated out-of-pocket costs for Medicare beneficiaries in each Part D prescription benefit phase, average retail price, total yearly costs and discounted prices through cash-pay discount coupons (GoodRx) or online pharmacies like Mark Cuban Cost Plus Drug Company (MCCPDC). We also report plan utilization management requirements. We analyzed 5721 plan formularies for 18 medications. Mirabegron was the only beta-3 agonist (B3). Only Vesicare oral solution (14.3% of plans) and Mirabegron (0.1%) required prior authorization. Many plans required step therapy for selective generic anticholinergics (ACH) (12.4%-43.3%), while the B3 rarely required step therapy (0.6%). Monthly costs varied by coverage phase and averaged $59 for ACHs in the initial coverage phase ($14 in catastrophic; $72 in coverage gap). The monthly cost for the B3 averaged $47 in the initial coverage phase ($26 in catastrophic; $129 in coverage gap). The total yearly cost for generic ACHs ranged from $494 (oxybutynin IR) to $1452 (darifenacin) and the yearly cost for brand-name ACHs ranged from $1175 (Toviaz ER) to $2198 (Oxytrol). The total yearly cost for the B3 was $1283. We evaluated coverage, out-of-pocket costs, total yearly costs, and utilization management for OAB medications to make pricing more transparent. While selective medications may be "covered," coverage does not translate into affordable drug prices.


Mouse haemoglobin beta chains. Comparative sequence data on adult major and minor beta chains from two species, Mus musculus and Mus cervicolor.
In inbred Mus musculus several different beta chains are known. In certain strains two beta chains are produced in unequal amounts by the two closely linked genes of the doublet breeding unit allele Hbb(d): Betadmaj and betadmin. One strain has a variant doublet allele, Hbb(p), which produces a variant minor beta chain, betapmin (the major beta chain, betapmaj, may not differ from betadmaj chain). Certain other strains have a singlet allele, Hbb(s), that produces only one beta chain, betas. Other species have different beta-chain patterns. In M. cervicolor two variant major beta chains are found, betacmaj (d-like) and betacmaj (s-like), both of which were found associated with minor beta chains. M. caroli has only one, 'Leporelike' beta chain, with structural features characteristic predominantly of betadmin chain in the N-terminal half and of betadmaj chain in the C-terminal half. The present paper presents sequence data on betas, betadmaj, betadmin, betapmin and betacmaj (d-like) chains. The data on betadmin chain cover almost the whole of that chain and show a minimum of nine differences from betadmaj chain and two from betapmin chain. It is suggested that the data on the beta chains of the various species show evidence for the past occurrence of double crossovers over regions within a gene coding for only one or a few amino acids, which events can be explained by the 'hybrid DNA' models of genetic recombination. Supplementary information on the amino acid sequence of the proteins has been deposited as Supplementary Publication SUP 50067 (36 pages) at the British Library Lending Division, Boston Spa, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1976) 153, 5.


Trans-sacral interbody fixation in long fusions to the sacrum for adult spinal deformity: complications and fusion rates at minimum two years follow-up.
This study aims to investigate the fusion rate and complications associated with trans-sacral interbody fusion (TSIF) in long fusions to the sacrum for adult spinal deformity (ASD) over a two year follow-up period. Potential predictor variables associated with pseudarthrosis were also examined. A retrospective clinical review was conducted on a consecutive series of ASD patients who underwent long fusions to the sacrum, with TSIF performed as a same-day or staged procedure. Patient demographics, bone mineral density, operative details, perioperative and late complications, and fusion rates were reviewed. Univariate analysis was used to identify the risk factors associated with pseudarthrosis. The study included 43 patients with an average age of 55.3 ± 8.9 years. The perioperative complication rate was 28%, with 12% of the complications directly related to TSIF. The late complication rate was 33%, with 16% related to TSIF. The most common complications were pseudarthrosis (14%) and postoperative ileus (7%). The overall radiographic fusion rate at two years was 86%. Univariate analysis revealed that revision surgery was significantly associated with pseudarthrosis (p = 0.027). Over the follow-up period, patients who underwent TSIF during long posterior fusions to the sacrum showed improvement in overall SRS scores, ODI scores, and SF-36 physical health and mental health (p < 0.05). TSIF is a relatively safe and minimally invasive method for achieving interbody fusion at the lumbosacral junction in the treatment of ASD, with acceptable fusion rates and a low complication rate. However, TSIF is not recommended for revision reconstruction in ASD.


Using an RNA aptamer probe for super-resolution imaging of native EGFR.
Aptamers, referred to as "chemical antibodies", are short single-stranded oligonucleotides that bind to targets with high affinity and specificity. Compared with antibodies, aptamers can be designed, developed and modified easily. Since their discovery, aptamers have been widely used in in vitro diagnostics and molecular imaging. However, they are relatively less studied and applied in advanced microscopy. Here we used an RNA aptamer in dSTORM imaging and obtained a high-quality image of EGFR nanoscale clusters on live cell membranes. The results showed that the cluster number and size with aptamer labeling were almost the same as those with labeling with the natural ligand EGF, but the morphology of the clusters was smaller and more regular than that with cetuximab labeling. Meanwhile, dual-color imaging demonstrated sufficient fluorophore labeling, highly specific recognition and greatly accurate clustering information provided by aptamers. Furthermore, the aptamer labeling method indicated that active EGFR formed larger clusters containing more molecules than resting EGFR, which was hidden under the antibody labeling. Our work suggested that aptamers can be used as versatile probes in super-resolution imaging with small steric hindrance, opening a new avenue for detailed and precise morphological analysis of membrane proteins.


Fusion rate following three- and four-level ACDF using allograft and segmental instrumentation: A radiographic study.
Cervical spine degenerative pathologies remain one of the most common spinal conditions treated by spine surgeons worldwide. Surgery is recommended in all patients with symptomatic cervical spinal stenosis with either moderate to severe myelopathy, degeneration, or refractory radiculopathy. As the number of levels increases the potential for complications associated with anterior surgery can be significant, especially dysphagia and pseudarthrosis. The objective of this study was to analyze the fusion rate following three- or more level anterior cervical discectomy and fusion (ACDF). A retrospective review was performed analyzing patients who underwent three or more level ACDF. Fusion was evaluated using post-operative dynamic upright radiographs Relevant post-operative complications especially dysphagia requiring dietary modifications or placement of feeding tube was also noted. A total of 72 patients were included in the study. Of the 232 levels fused, pseudarthrosis occurred at 47 (14%) levels. Overall 45.8% of patients (33/72) had a pseudarthrosis. The incidence of pseudarthrosis was higher in patients with 4 level ACDF as compared to those with 3 level ACDF [56% (9/16) versus 42% (24/56)]. At last follow up, the number of patients that were symptomatic from their pseudarthrosis and required posterior spinal instrumentation was 8/72 (11.1%). Fusion rates in a large cohort of patients with three- and four-level ACDF performed utilizing allograft and segmental instrumentation is reported. The study demonstrates that 3-4 level, stand-alone anterior cervical arthrodeses result in at least one level of pseudarthrosis in almost half of patients, especially at the caudal level of the construct.


Hydraulic testing of intravascular axial flow blood pump designs with a protective cage of filaments for mechanical cavopulmonary assist.
To provide hemodynamic support to patients with a failing single ventricle, we are developing a percutaneously inserted, magnetically levitated axial flow blood pump designed to augment pressure in the cavopulmonary circulation. The device is designed to serve as a bridge-to-transplant, bridge-to-recovery, bridge-to-hemodynamic stability, or bridge-to-surgical reconstruction. This study evaluated the hydraulic performance of three blood pump prototypes (a four-bladed impeller, a three-bladed impeller, and a three-bladed impeller with a four-bladed diffuser) whose designs evolved from previous design optimization phases. Each prototype included the same geometric protective cage of filaments, which stabilize the rotor within the housing and protect the housing wall from the rotating blades. All prototypes delivered pressure rises over a range of flow rates and rotational speeds that would be sufficient to augment hemodynamic conditions in the cavopulmonary circulation. The four-bladed impeller outperformed the two remaining prototypes by >40%; this design was able to generate a pressure rise of 4-28 mm Hg for flow rates of 0.5-10 L/min at rotational speeds of 4,000-7,000 RPM. Successful development of this blood pump will provide clinicians with a feasible therapeutic option for mechanically supporting the failing Fontan.


A goal set too high: factors associated with planning excess gestational weight gain in a prospective cohort study.
This study aimed to understand physical, knowledge, psychological, and lifestyle factors associated with planned excess gestational weight gain (GWG), a strong and potentially modifiable predictor of actual excess GWG, which contributes to maternal and child obesity along with other adverse maternal and fetal outcomes. This is a secondary analysis of data from a prospective cohort study where women completed a questionnaire in early pregnancy. Women were asked to report their planned GWG, which was then categorized as above, within, or below the Institute of Medicine (IOM) guidelines. Univariable and multivariable analyses were performed to identify variables associated with planned excess GWG. Of 970 women included in the analysis, 300 reported a planned GWG above the IOM guidelines. Predictors of excess planned GWG included reporting healthcare provider recommendations to gain weight above the guidelines (adjusted odds ratio [aOR], 62.17; 95% confidence interval [CI], 13.75-281.03), overestimating first trimester weight gain recommendations (aOR, 1.83; 95% CI, 1.21-2.77), believing in risks to the baby with inadequate GWG (aOR 2.16; 95% CI,1.29-3.60), inaccurate self-perceived prepregnancy body size (aOR, 1.88; 95% CI, 1.22-2.89), low or high emotional suppression (aOR, 1.78; 95% CI, 1.06-2.99; and aOR, 2.57; 95% CI, 1.21-5.45, respectively), physical inactivity (aOR, 1.10; 95% CI, 1.03-1.17), and overweight or obesity (aOR, 5.76; 95% CI, 3.70-8.98; and aOR, 11.46; 95% CI, 6.54-20.06, respectively). Protective factors against planned excess GWG included increased maternal age (aOR, 0.95; 95% CI, 0.92-0.99), and believing in risks to themselves with inadequate GWG (aOR 0.64; 95% CI, 0.42-0.97) or believing in risks to the baby with excess GWG (aOR, 0.49; 95% CI, 0.27-0.88). Women with overweight or obesity are at greater risk of prospectively planning excess GWG, and may especially benefit from healthcare provider counseling on appropriate GWG. Other modifiable factors for planned excess GWG included knowledge about risks of inappropriate weight gain and physical inactivity.


Allometric scaling of foraging rate with trail dimensions in leaf-cutting ants.
Leaf-cutting ants (Atta spp.) create physical pathways to support the transport of resources on which colony growth and reproduction depend. We determined the scaling relationship between the rate of resource acquisition and the size of the trail system and foraging workforce for 18 colonies of Atta colombica and Atta cephalotes. We examined conventional power-law scaling patterns, but did so in a multivariate analysis that reveals the simultaneous effects of forager number, trail length and trail width. Foraging rate (number of resource-laden ants returning to the nest per unit time) scaled at the 0.93 power of worker numbers, the -1.02 power of total trail length and the 0.65 power of trail width. These scaling exponents indicate that individual performance declines only slightly as more foragers are recruited to the workforce, but that trail length imposes a severe penalty on the foraging rate. A model of mass traffic flow predicts the allometric patterns for workforce and trail length, although the effect of trail width is unexpected and points to the importance of the little-known mechanisms that regulate a colony's investment in trail clearance. These results provide a point of comparison for the role that resource flows may play in allometric scaling patterns in other transport-dependent entities, such as human cities.


Axon-sparing lesions of the medial nucleus of the amygdala decrease affiliative behaviors in the prairie vole (Microtus ochrogaster): behavioral and anatomical specificity.
The neural basis of affiliative behavior was examined in the prairie vole, a rodent that exhibits high levels of social contact and paternal behavior. In the first study, the axon-sparing excitotoxin N-methyl-D,L-aspartic acid (NMA) produced lesions in the basolateral nucleus of the amygdala or the corticomedial amygdala. Males with corticomedial lesions showed significantly less contact with a familiar adult female and a pup when compared with males with lesions of the basolateral nucleus or controls. This behavioral change was not associated with changes in exploratory behavior, motor function, performance in an olfactory task, fearfulness, physical well-being, or body temperature. In a second study, NMA lesions restricted to the medial nucleus also decreased paternal behavior. Neurons in the medial nucleus of the amygdala appear to be essential for the normal expression of paternal care in this species.


Charged microcapsules for controlled release of hydrophobic actives. Part III: the effect of polyelectrolyte brush- and multilayers on sustained release.
Poly(methyl methacrylate) microspheres have been prepared by the internal phase separation method using either of the three conventional dispersants poly(vinyl alcohol) (PVA), poly(methacrylic acid) (PMAA), or the amphiphilic block copolymer poly(methyl methacrylate)-block-poly(sodium methacrylate). The block copolymer based microsphere, which has a polyelectrolyte brush on the surface, was surface modified with up to two poly(diallyldimethylammonium chloride)-poly(sodium methacrylate) bilayers. The microspheres were loaded with the hydrophobic dye 2-(4-(2-chloro-4-nitrophenylazo)-N-ethylphenylamino)ethanol (Disperse Red 13) and its release from aqueous dispersions of microspheres with the different surface compositions was measured by spectrophotometry. The burst fraction, burst rate and the diffusion constant were determined from a model combining burst and diffusive release. Out of the three dispersants, the block copolymer gave the slowest release of the dye, with respect to both burst release and diffusive release. A very pronounced further reduction of the diffusion constant was obtained by applying polyelectrolyte multilayers on top of the microspheres. However, the diffusion constant was very weakly dependent on further polyelectrolyte adsorption and one polyelectrolyte bilayer appeared to suffice.


Longitudinal monitoring of extended-spectrum-beta-lactamase/AmpC-producing Escherichia coli at German broiler chicken fattening farms.
Antimicrobial resistance of Escherichia coli to modern beta-lactam antibiotics due to the production of extended-spectrum beta-lactamases (ESBL) and/or plasmid-mediated AmpC beta-lactamases (AmpC) represents an emerging and increasing resistance problem that dramatically limits therapeutic options in both human and veterinary medicine. The presence of ESBL/AmpC genes in commensal E. coli from food-producing animals like broilers may pose a human health hazard. However, there are no data available concerning the prevalence of ESBL/AmpC-producing E. coli in German broiler flocks using selective methods. In this longitudinal study, samples were taken from seven conventional broiler fattening farms at three different times within one fattening period. Various samples originating from the animals as well as from their direct environment in the barn were investigated for the occurrence of ESBL/AmpC-producing E. coli. Average detection levels of 51, 75, and 76% in animal samples collected during the three samplings in the course of the fattening period demonstrate a colonization of even 1-day-old chicks, as well as a continuous significant (P < 0.001) increase in prevalence thereafter. The detection frequencies in housing environmental samples were relatively high, with an increase over time, and ranged between 54.2 and 100%. A total of 359 E. coli isolates were characterized by PCR and partly via the disc diffusion method. This study shows that prevalence of ESBL/AmpC-producing E. coli increases during the fattening period of the broiler flocks examined. Both colonized day-old chicks and contaminated farm environments could represent significant sources of ESBL/AmpC-producing E. coli in German broiler fattening farms.


Mediators of the effectiveness of a kindergarten-based, family-involved intervention on pre-schoolers' snacking behaviour: the ToyBox-study.
The present study aimed to explore the mediating role of family-related determinants on the effects of the ToyBox-intervention on pre-school children's consumption of healthy and unhealthy snacks. The ToyBox-intervention was a kindergarten-based, family-involved intervention with a cluster-randomized design, aiming to promote healthy lifestyle behaviours to prevent obesity at pre-school age. Kindergartens (n 309) in six European countries. A total of 6290 pre-schoolers and their families participated in the ToyBox-intervention in 2012-2013 and data from 5212 pre-schoolers/families were included in the current analyses. Even though the total effect of the ToyBox-intervention on healthy and unhealthy snacking was not significant, the ToyBox-intervention significantly improved parental rule setting on children's unhealthy snack consumption (i.e. restriction of snacking while watching television and permission only at certain occasions) and parental consumption of unhealthy snacks, while it increased parental knowledge on snacking recommendations. Regarding healthy snacking, the ToyBox-intervention improved children's attitude towards fruit and vegetables (F&V). All previously mentioned family-related determinants mediated the intervention effects on pre-schoolers' consumption of healthy and unhealthy snacks. Almost all family-related determinants examined in the study were independently associated with pre-schoolers' consumption of healthy and unhealthy snacks. The intervention was effective in improving relevant family-related determinants. Interventions aiming to promote F&V consumption and limit the consumption of unhealthy snacks in pre-schoolers should target on these mediators, but also identify new family-, school- or peer-related determinants, to enhance their effectiveness.


Epidemiology of suicide among children and adolescents in Austria, 2001-2014.
Previous epidemiological analyses indicated a decreasing trend of suicide rates for 10-19-year-olds in Austria for the period 1970-2001. However, data from the new millennium are missing. This epidemiological update reports on youth suicide in Austria, covering the period 2001-2014 in order to inform suicide preventive interventions targeting adolescents. The data on registered suicides among Austrian minors (10-19 years) and the population size were obtained from Statistics Austria. Chi-squared tests were used to analyze the associations between the suicide methods used and sex, as well as between suicide methods and Austrian federal states. Spearman correlations were calculated to assess time trends in the suicide rates. One-way ANOVA was used to investigate annual suicide rates of age groups 10-14, 15-19, and 10-19 years across the nine Austrian federal states. The total average suicide rate for Austrian minors was 4.57 per 100,000. The male-female ratio was 3.5:1. The total youth suicide rate and male suicide rate significantly declined from 2001 to 2014, whereas there were no significant changes in female rates. More than one third of suicides among Austrian youth occurred through hanging, whereas jumping in front of a moving object was the second-most common suicide method. A spring peak was found, with most suicides occurring in April and May. Suicide rates among minors in Austria continue to decrease. The present findings help to inform the ongoing implementation of the National Austrian Suicide Prevention Plan (SUPRA).


The forgotten smoker: a qualitative study of attitudes towards smoking, quitting, and tobacco control policies among continuing smokers.
Although research suggests that the majority of smokers want to quit smoking, the uptake of Stop Smoking Services, designed to assist smokers with quitting, remains low. Little is known about continuing smokers who do not access these services, and opportunities to influence their motivation and encourage quit attempts through the uptake of services. Using PRIME theory, this study explored differences between continuing smokers who had varying levels of motivation to quit, in terms of their plans to quit, evaluative beliefs about smoking, cigarette dependence, and attitudes towards tobacco control policies and services. Twenty-two current smokers, recruited from the community, were classified by motivation level to quit using a self-report questionnaire (two groups: high/low). Four focus groups (n=13) and individual interviews (n=9) were conducted with both groups using an interview guide incorporating aspects of PRIME theory. Discussion areas included motives for smoking, attitudes towards smoking and quitting, perceptions of dependence, motives for quitting, barriers to quitting, and attitudes towards existing and impending tobacco control policies and services. Verbatim transcripts were analysed using thematic framework analysis. All participants expressed low motivation to quit during discussions, despite some initially self-classifying as having high explicit levels of motivation to quit. Both groups reported similar attitudes towards smoking and quitting, including a perceived psychological addiction to smoking, positive evaluations about smoking which inhibited plans to quit, and similar suggested methods to increase motivation (simply wanting to, save money, improve health). Most felt that they 'ought' to quit as opposed to 'wanted' to. Little influence was ascribed towards tobacco control policies such as plain packaging and hidden sales displays, and participants felt that price increases of tobacco products needed to be considerable in order to influence motivation. Highly motivated smokers expressed more willingness to visit Stop Smoking Services, although none had done so. Continuing smokers' attitudes towards smoking and quitting suggests that research and policy need to focus on increasing smokers' implicit motivation to quit smoking, even for those who classified themselves as having high motivation to quit. Targeted information and further education about Stop Smoking Services is required to increase uptake.


Two distinct domains of the glucagon-like peptide-1 receptor control peptide-mediated biased agonism.
G protein-coupled receptors (GPCRs) can be differentially activated by ligands to generate multiple and distinct downstream signaling profiles, a phenomenon termed biased agonism. The glucagon-like peptide-1 receptor (GLP-1R) is a class B GPCR and a key drug target for managing metabolic disorders; however, its peptide agonists display biased signaling that affects their relative efficacies. In this study, we combined mutagenesis experiments and mapping of surface mutations onto recently described GLP-1R structures, which revealed two major domains in the GLP-1/GLP-1R/Gs protein active structure that are differentially important for both receptor quiescence and ligand-specific initiation and propagation of biased agonism. Changes to the conformation of transmembrane helix (TM) 5 and TM 6 and reordering of extracellular loop 2 were essential for the propagation of signaling linked to cAMP formation and intracellular calcium mobilization, whereas ordering and packing of residues in TMs 1 and 7 were critical for extracellular signal-regulated kinase 1/2 (pERK) activity. On the basis of these findings, we propose a model of distinct peptide-receptor interactions that selectively control how these different signaling pathways are engaged. This work provides important structural insight into class B GPCR activation and biased agonism.


CXC-chemokine/CXCR2 biological axis promotes angiogenesis in vitro and in vivo in pancreatic cancer.
Angiogenesis is essential for tumor growth and metastasis. Although ELR(+)-CXC-chemokines and their corresponding receptor, CXC-receptor 2 (CXCR2), are known mediators of angiogenesis, little is known about their role in pancreatic cancer (PaCa). The aim of our study was to determine the role of ELR(+)-CXC-chemokine/CXCR2 biological axis in promoting PaCa angiogenesis. We prospectively collected secretin-stimulated exocrine pancreatic secretions (SSEPS) from normal individuals (NP) and PaCa patients. We showed that summed concentrations of ELR(+)-CXC-chemokines in SSEPS from PaCa patients were significantly higher than in those from NP (p = 0.002). We measured ELR(+)-CXC-chemokine levels in supernatants from multiple PaCa cell lines and confirmed that BxPC-3, Colo-357 and Panc-28 had significantly higher expression compared with an immortalized human pancreatic ductal epithelial (HPDE) cell line. After confirming lack of autocrine effects of ELR(+)-CXC-chemokines on PaCa cells (due to absence of CXCR2 expression), we investigated paracrine effects of these chemokines on human umbilical vein endothelial cells (HUVEC). Both recombinant ELR(+)-CXC-chemokines and co-culturing with BxPC-3 significantly enhanced proliferation, invasion, and tube formation of HUVEC (p < 0.05). These biological effects were significantly inhibited by treatment with a neutralizing antibody against CXCR2 (anti-CXCR2 Ab) (p < 0.05). Finally, anti-CXCR2 Ab significantly reduced tumor volume (p < 0.05), Ki-67 proliferation index (p = 0.043) and Factor VIII(+) microvessel density (p = 0.004) in an orthotopic nude mouse PaCa model. Our results show that ELR(+)-CXC-chemokines promote PaCa tumor-associated angiogenesis through CXCR2, suggesting that CXCR2 is an anti-angiogenic target in PaCa.


Acute myeloid leukemia (AML) with t(7;12)(q36;p13) is associated with infancy and trisomy 19: Data from Nordic Society for Pediatric Hematology and Oncology (NOPHO-AML) and review of the literature.
The t(7;12)(q36;p13) (MNX1/ETV6) is not included in the WHO classification but has been described in up to 30% of acute myeloid leukemia (AML) in children <2 years and associated with a poor prognosis. We present the clinical and cytogenetics characteristics of AML cases with t(7;12)(p36;p13). A literature review identified 35 patients with this translocation, published between 2000 and 2015. Outcome data were available in 22 cases. The NOPHO-AML (Nordic Society for Pediatric Hematology and Oncology) database contained 651 patients with AML from 1993 to 2014 and seven (1.1%) had the translocation. The t(7;12) was only present in patients <2 years of age (median age 6 months) but none was diagnosed as newborn. These patients constituted 4.3% of the patients <2 years of age. There was a strong association with trisomy 19 (literature: 86%, NOPHO: 100%) and +8 (literature: 19%, NOPHO: 14%). Seventeen of 22 patients from the literature with t(7;12) and four of seven patients from the NOPHO database suffered from relapse. The patients with t(7;12) had a 3-year event free survival of 24% (literature) vs. 43% (NOPHO) and a 3-year overall survival of 42% (literature) vs. 100% (NOPHO). None of the NOPHO patients was treated with hematopoietic stem cell transplantation (HSCT) in first complete remission. Relapse was frequent but the salvage rate using HSCT was high. We conclude that t(7;12)(q36;13) is a unique subgroup of childhood AML with presentation before 2 years of age with most cases being associated with +19.


Cannabinoids induce latent sensitization in a preclinical model of medication overuse headache.
Evaluation of cannabinoid receptor agonists in a preclinical model of medication overuse headache. Female Sprague Dawley rats received graded intraperitoneal doses of WIN55,212-2 or Δ-9-tetrahydrocannabinol (Δ-9-THC). Antinociception (tail-flick test), catalepsy and hypomotility (open field test) and impairment of motor function (rotarod test) were assessed to establish effective dosing. Rats were then treated twice daily with equianalgesic doses of WIN55,212-2 or Δ-9-THC, or vehicle, for 7 days and cutaneous tactile sensory thresholds were evaluated during and three weeks following drug discontinuation. Rats then received a one-hour period of bright light stress (BLS) on two consecutive days and tactile sensory thresholds were re-assessed. WIN55,212-2 and Δ-9-THC produced antinociception as well as hypomotility, catalepsy and motor impairment. Repeated administration of WIN55,212-2 and Δ-9-THC induced generalized periorbital and hindpaw allodynia that resolved within 3 weeks after discontinuation of drug. Two episodes of BLS produced delayed and long-lasting periorbital and hindpaw allodynia selectively in rats previously treated with WIN55,212-2, and Δ-9-THC. Cannabinoid receptor agonists including Δ-9-THC produce a state of latent sensitization characterized by increased sensitivity to stress, a presumed migraine trigger. Overuse of cannabinoids including cannabis may increase the risk of medication overuse headache in vulnerable individuals.


Genomic structure of mouse SPI-C and genomic structure and expression pattern of human SPI-C.
Erythroblast transformation-specific domain (ETS) transcription factors regulate some of the critical molecular mechanisms controlling the differentiation of multipotent haematopoietic progenitor cells into effector B-lymphocytes. The SPI-group ETS-protein transcription factors PU.1 and SPI-B play essential and, although coexpressed and binding to similar DNA sequences, unique roles in B-cell differentiation in mice. Mouse SPI-C is an SPI-group ETS protein expressed temporarily during B-cell development and in macrophages. Here we present the genomic organization of the mouse SPI-C gene, and show by rapid amplification of cDNA ends (5'-RACE) analysis that transcription of the mouse SPI-C mRNA starts at a single site producing a single processed transcript. We have also isolated a cDNA clone encoding the human SPI-C homologue, which displays 65% amino acid identity to the murine protein. In addition, we show that the genomic structure of the human and mouse genes are similar, containing a 5' non-coding exon followed by five coding exons. Human SPI-C mRNA is preferentially detected in foetal and adult spleen, lymph nodes and at lower levels in bone marrow and foetal liver. Finally a phylogenetic prediction analysis of SPI-group protein sequences suggest that the SPI-C proteins form a distinct subgroup, with human SPI-C being closest related to the mouse SPI-C protein.


Cdx1 and Cdx2 exhibit transcriptional specificity in the intestine.
The caudal-related homeodomain transcription factors Cdx1 and Cdx2 are expressed in the developing endoderm with expression persisting into adulthood. Cdx1(-/-) mutants are viable and fertile and display no overt intestinal phenotype. Cdx2 null mutants are peri-implantation lethal; however, conditional mutation approaches have revealed that Cdx2 is required for patterning the intestinal epithelium and specification of the colon. Cdx2 is also necessary for homeostasis of the intestinal tract in the adult, where Cdx1 and Cdx2 appear to functionally overlap in the distal colon, but not during intestinal development. Cdx1 and Cdx2 exhibit complete overlap of expression in the intestine, although they differ in their relative levels, with Cdx1 maximal in the distal colon and Cdx2 peaking in the proximal cecum. Moreover, Cdx1 protein is graded along the crypt-villus axis, being abundant in the crypts and diminishing towards the villi. Cdx2 is expressed uniformly along this axis, but is differentially phosphorylated; the functional relevance of these expression domains and phosphorylation is currently unknown. Cdx1 and Cdx2 have been suggested to exhibit functional specificity in the intestinal tract. In the present study, using cell-based models, we found that relative to Cdx1, Cdx2 was significantly less potent at effecting a transcriptional response from the Cdx1 promoter, a known Cdx target gene. We subsequently assessed this relationship in vivo using a "gene swap" approach and found that Cdx2 cannot substitute for Cdx1 in this autoregulatory loop. This is in marked contrast with the ability of Cdx2 to support Cdx1 expression and function in paraxial mesoderm and vertebral patterning, thus providing novel in vivo evidence of context-dependent transcriptional specificity between these transcription factors.


Heat-induced conformational unfolding of fatty acid-free and fatted camel serum albumin: a comparative study.
Albumin is a multifunctional non-glycosylated, negatively charged plasma protein, with extraordinary ligand-binding and transport properties, antioxidant functions, and enzymatic activities. Physiologically, albumin transports free fatty acids in plasma and contributes in maintaining colloid osmotic pressure. Recent progresses in using albumin as a versatile protein carrier for drug targeting and for improving the pharmacokinetic profile of peptide or protein-based drugs, increased the attempts for improving albumin stability. Studying the thermal stability of camel albumin may provide us not only new clues for designing recombinant albumins, but also molecular insights on camel physiology. This study aims to determine the thermal stability of camel albumin. Fatted camel serum albumin (FCSA) was purified from blood via combination of Cohn's method and anion-exchange chromatography. Activated charcoal treatment was used to obtain defatted camel serum albumin (CSA). Fluorescence spectroscopy and differential scanning calorimetry (DSC) were used to study thermal denaturation of this protein. The set of fluorescence spectra were deconvoluted using the convex constraint analysis method (CCA). The results from deconvolution of fluorescence spectroscopy and DSC showed three and two components for CSA and FCSA, respectively. The bimodal DSC transition can be attributed to a crevice between domains I and II and formation of two independent thermodynamic domains. The crevice formation can be prevented by fatty acid binding between domains I and II. The calculated values of ∆H v/∆H cal, approximately 0.4 for CSA and near 1 for FCSA, confirmed the presence of at least one intermediate in thermal unfolding of CSA and the absence of the intermediate for FCSA. The obtained midpoint transition temperature (T m) of FCSA was about 20 °C higher than that of CSA. Such enormous stabilizing effect may be attributed to the fact that fatty acid serves as glue which preserves different domains beside each other and prevents formation of the mentioned intermediate.


50W vs 40W During High-Power Short-Duration Ablation for Paroxysmal Atrial Fibrillation: A Multicenter Prospective Study.
High-power short-duration (HPSD) radiofrequency ablation of atrial fibrillation (AF) increases first-pass pulmonary vein isolation (PVI) and freedom from atrial arrhythmias while decreasing procedural time. However, the optimal power setting in terms of safety and efficacy has not been determined. This study compared the procedural characteristics and clinical outcomes of 50W vs 40W during HPSD ablation of paroxysmal AF. Patients from the REAL-AF prospective multicenter registry (Real-World Experience of Catheter Ablation for Treatment of Symptomatic Paroxysmal and Persistent Atrial Fibrillation) undergoing HPSD ablation of paroxysmal AF, either using 50W or 40W, were included. The primary efficacy outcome was freedom from all-atrial arrhythmias. The primary safety outcome was the occurrence of any procedural complication at 12 months. Secondary outcomes included procedural characteristics, AF-related symptoms, and the occurrence of transient ischemic attack or stroke at 12 months. A total of 383 patients were included. Freedom from all-atrial arrhythmias at 12 months was 80.7% in the 50W group and 77.3% in the 40W group (Log-rank P = 0.387). The primary safety outcome occurred in 3.7% of patients in the 50W group vs 2.8% in the 40W group (P = 0.646). The 50W group had a higher rate of first-pass PVI (82.3% vs 76.2%; P = 0.040) as well as shorter procedural (67 minutes [IQR: 54-87.5 minutes] vs 93 minutes [IQR: 80.5-111 minutes]; P < 0.001) and radiofrequency ablation times (15 minutes [IQR: 11.4-20 minutes] vs 27 minutes [IQR: 21.5-34.6 minutes]; P < 0.001) than the 40W group. There was no significant difference in freedom from all-atrial arrhythmias or procedural safety outcomes between 50W and 40W during HPSD ablation of paroxysmal AF. The use of 50W was associated with a higher rate of first-pass PVI as well as shorter procedural times.


[Isolation of anti-Cryptococcus neoformans immunoglobulin G].
Two immunization schemes were applied to rabbits with an autoctonous strain aimed at obtaining an anti-Crytococcus neoformans IgG. Whole cells treated with formalin were used. Dosages and immunization routes were different. The sera obtained were titered and that with the highest titer was selected (1:1 024 by the laminar agglutination method) and purified by saline precipitation and ion exchange chromatography. An IgG of high purity and biological activity was obtained.


Cytoplasmic Maspin Expression Correlates with Poor Prognosis of Patients with Adenocarcinoma of the Uterine Cervix.
Maspin is known to be a tumor suppressor protein and its prognostic significance in patients with several types of cancer has been reported. To date, however, no study has focused on the association between maspin expression and the prognosis of patients with adenocarcinoma of the uterine cervix. We explored the prognostic value of maspin expression with particular reference to its subcellular localization in patients with adenocarcinoma of the uterine cervix. Paraffin-embedded tissue samples from 46 patients diagnosed as adenocarcinoma of the uterine cervix were immunohistochemically analyzed using an antibody for maspin. The patients were followed up for 3 to 165 months (median: 64.2 months) and the prognostic value was evaluated by the log-rank test and the Cox regression hazard model. A sample was considered maspin-positive if maspin was expressed in only the cytoplasm; 69.6% (32 cases) of the specimens were maspin-positive, and there was significant correlation between positivity and recurrence (P = 0.022). Maspin-positive patients had both shorter disease free survival and shorter overall survival by the log-rank test (P = 0.023, P = 0.043, respectively). By Cox's multivariate analysis, the International Federation of Obstetrics and Gynecology (FIGO) status was the only independent prognostic factor for disease free survival and overall survival in patients with adenocarcinoma of the uterine cervix. This is the first report to reveal an association between cytoplasmic maspin expression and the prognosis of patients with adenocarcinoma of the uterine cervix. Although further studies with a larger series of patients and a longer follow up period are necessary, the present results suggest that cytoplasmic maspin expression could be an indicator of unfavorable prognosis in patients with adenocarcinoma of the uterine cervix.


Radioactivity of building materials and the gamma radiation in dwellings.
Measurements of the radioactivity in some common building materials in Norway are reported, together with calculations of the gamma-ray exposure from walls of different materials. Model rooms are used in calculations of the mean exposure inside concrete, brick and light-weight expanded clay aggregate buildings. These calculations give very good agreement with previous experimental results. The radiological implications of using building materials with high concentrations of radioactivity are also discussed.


Impact of Image Reconstruction Parameters on Abdominal Aortic Calcification Measurement Using Abdominal Computed Tomography.
Abdominal aortic calcification (AAC) is correlated with cardiovascular outcomes independent of traditional risk factors. Quantification of AAC on computed tomography (CT) has not been standardized. Reconstruction parameters have been shown to impact coronary calcium scores. The aim of our study was to assess the impact of abdominal CT reconstruction parameters, slice thickness (ST), and display field of view (DFOV) on AAC quantitative scoring on abdominal CT examinations. We retrospectively measured AAC on noncontrast CT of 46 patients (mean age, 64.1 years; 35 males) using 5 different reconstruction protocols with a range of ST and DFOV: protocol A, 2.5 mm ST, 35 cm DFOV; protocol B, 2.5 mm ST, 50 cm DFOV; protocol C, 2.5 mm ST, 25 cm DFOV; protocol D, 5 mm ST, 35 cm DFOV; and protocol E: 0.625 mm ST, 35 cm DFOV. The AAC scores from each protocol were compared using concordance correlation coefficient and Bland-Altman agreement analyses. The AAC mean (SD) scores for each protocol were as follows: A, 2022 (2418); B, 2022 (2412); C, 1939 (2310); D, 2220 (2695); and E, 1862 (2234). The AAC mean score differences between protocols and reference protocol A were -0.47, 82.01, -198.94, and 160 for protocols B, C, D, and E, respectively, with differences between protocols C to E statistically significantly different (P < 0.05). The different protocols showed overall excellent correlation (concordance correlation coefficient, >0.9) between AAC scores. Slice thickness and DFOV can impact AAC score measurement. A description of reconstruction parameters is important to allow comparisons across different cohorts.


Phylogenetic conservatism of mycoparasitism and its contribution to pathogen antagonism.
Closely related species are expected to have similar functional traits due to shared ancestry and phylogenetic inertia. However, few tests of this hypothesis are available for plant-associated fungal symbionts. Fungal leaf endophytes occur in all land plants and can protect their host plant from disease by a variety of mechanisms, including by parasitizing pathogens (e.g., mycoparasitism). Here, we tested whether phylogenetic relatedness among species of Cladosporium, a widespread genus that includes mycoparasitic species, predicts the effect of this endophyte on the severity of leaf rust disease. First, we used congruence among different marker sequences (i.e., genealogical concordance phylogenetic species recognition criterion) to delimit species of Cladosporium. Next, in a controlled experiment, we quantified both mycoparasitism and disease modification for the selected Cladosporium species. We identified 17 species of Cladosporium; all the species reduced rust disease severity in our experiment. Cladosporium phylogeny was a significant predictor of mycoparasitism. However, we did not observe a phylogenetic effect on disease severity overall, indicating that other mechanism/s operating independently of shared ancestry also contributed to endophyte effects on disease severity. Indeed, a second experiment showed that Cladosporium endophyte exudates (no live organism) from divergent species groups equally reduced disease severity. Our results reveal that multiple mechanisms contribute to the protective effects of an endophyte against a plant pathogen, but not all traits underlying these mechanisms are phylogenetically conserved.


L-[U-14C] lactate binding to a 43 kDa protein in plasma membranes of Candida utilis.
To identify the putative lactate transporter protein of Candida utilis, plasma membranes from cells grown either on lactic acid (presence of lactate proton symport) or glucose (absence of lactate proton symport) were incubated with L-[U-14C]lactic acid and the membrane proteins were then separated by SDS-PAGE. A well-defined peak of radioactivity occurred in the lane of the gel containing plasma membrane proteins from lactic-acid-grown cells but not from glucose-grown cells. Binding was inhibited by unlabelled pyruvate and lactate, whereas succinate and citrate were not inhibitory. The monocarboxylate transporter inhibitor of animal cells, 4,4'-diisothiocyanatostilbene-2,2'-disulfonate, competitively inhibited the lactate proton symport in the whole yeast and also inhibited lactate binding to proteins of isolated plasma membranes. The polypeptide pattern of plasma membranes from lactic-acid-grown cells revealed a 43 kDa polypeptide associated with the peak of labelled lactate. Altogether the results suggest that this polypeptide is either the lactate transporter or a component of it.


Automated dispensing cabinet alert improves compliance with obtaining blood cultures before antibiotic administration for patients admitted with pneumonia.
A Centers for Medicare & Medicaid Services (CMS) pneumonia quality measures with particular impact on the emergency department (ED) is blood cultures prior to antibiotic administration for patients admitted with pneumonia. A study was conducted to measure the impact of an automated dispensing cabinet (ADC) alert on improving compliance with the quality measure of obtaining blood cultures prior to giving antibiotics for patients admitted with pneumonia and who have blood cultures ordered. The pre-post study involved ED adult patient with an admitting diagnosis of pneumonia from October 2007 through September 2008. The intervention consisted of a series of questions in the ED medication ADC regarding blood culture orders and antibiotic administration. Patients with an admitting diagnosis of pneumonia were identified through a search of the ED electronic health record (EHR). The proportion of patients in whom blood cultures were obtained prior to antibiotic administration in the pre- (October 2007-March 2008) and postintervention (April 2008-September 2008) periods were compared. The chi-square test was used to test for statistical significance. Some 951 patients with pneumonia were identified during the study period, 426 pre- and 525 postintervention. Compliance with obtaining blood cultures prior to antibiotic administration was 84% (205/245, 95% confidence interval [CI]: 79%-88%) and 95% (275/291, 95% CI: 92%-97%) in the pre and postintervention periods, respectively (p <. 001). In this population of patients with pneumonia, a series of questions in an ADC improved compliance with the quality measure regarding the obtaining of blood cultures prior to administering antibiotics to patients in whom blood cultures are requested.


Compassion meditators show less anger, less punishment, and more compensation of victims in response to fairness violations.
Fairness violations elicit powerful behavioral and affective responses. Indeed, people are willing to incur costs to sanction unfair behavior. Here we study the possible impact of long-term mental training in socio-affective capacities such as compassion on altruistic punishment and compensatory behavior in economic games. To this end we recruited a group of long-term meditation practitioners (LTPs) who had engaged in an average of 40 K h of mental training exercises including compassion-related meditation, along with a group of meditation-naïve controls. Participants played several adaptations of the dictator game in which they had the opportunity to punish the dictator both when they were the recipients of the dictator's offer and when they were third-party witnesses to the dictator's treatment of an anonymous second player. Compared to controls, LTPs were less likely to punish when they were the victims of fairness violations. However, both groups punished equivalently when they witnessed others receiving unfair treatment. In post-task questionnaires, controls reported significantly more anger in response to unfair offers than LTPs, although fairness judgments did not differ between groups. These data suggest that because the LTPs were less angered by unfair treatment of themselves, they punished that behavior less. However, when they witnessed the unfair treatment of others, they engaged in norm-reinforcing punishment. Finally, when participants played an additional game which included the opportunity to recompense victims, LTPs were more likely to do so. Together these data point to differential approaches to justice whereby LTPs engaged less in vengeful, retributive justice and focused more on norm reinforcement and the restoration of equity. These differences suggest that social preferences are plastic and that altruistic responses to unfairness may be shaped by the prolonged cultivation of prosocial motivation, altruism, and compassion.


The rate of breast-conserving surgery for early breast cancer is not influenced by the surgical strategy of excisional biopsy followed by the definitive procedure.
Increased emphasis on breast conservation and the primacy of the patient's preferences has led to the promotion and increased use of a two-step surgical strategy (definitive operation only after a final tissue diagnosis from a biopsy done on a previous visit) in the treatment of early breast cancer, with the assumption being that this is more conducive to the performance of breast-conserving surgery (BCS). We sought to test this by examining the effect of the surgical strategy (one-step versus two-step) on the operation performed (BCS versus mastectomy). A random sample of women with node-negative breast cancer diagnosed in 1991 in Ontario was drawn from the Ontario Cancer Registry database and matched to the Canadian Institute of Health Information and Ontario Health Insurance Plan databases (n = 643). This provided information on the timing and nature of all surgical procedures performed as well as patient, tumor, hospital, and surgeon characteristics. The surgical strategy was defined as either a one-step procedure (biopsy and definitive surgery performed at the same time) or a two-step procedure (surgical biopsy and pathologic diagnosis, followed by definitive surgery at a later date). The axillary lymph node dissection was used to define the definitive procedure. BCS was employed in 68% of patients, and this did not differ significantly between the one-step and two-step groups (66% versus 70%). Patients with palpable lesions had a significantly lower rate of breast conservation than those with nonpalpable lesions. Other variables associated with a lower rate of BCS were larger tumor size, presence of extensive ductal carcinoma in situ (DCIS), and central or multifocal tumors. The use of a one-step procedure was associated with a patient age of more than 50 years, a palpable mass, tumor size larger than 1 cm, previous fine needle aspiration (FNA) biopsy, absence of extensive DCIS, and surgery in an academic setting. Breast conservation was not affected by the surgical strategy used or the timing of the decision, but was associated with several accepted tumor factors. This study shows that, contrary to the opinion of some, there is a group of breast cancer patients in whom treatment in a one-step manner is appropriate.


Critical role of retinoid/rexinoid signaling in mediating transformation and therapeutic response of NUP98-RARG leukemia.
While the nucleoporin 98-retinoic acid receptor gamma (NUP98-RARG) is the first RARG fusion protein found in acute leukemia, its roles and the molecular basis in oncogenic transformation are currently unknown. Here, we showed that homodimeric NUP98-RARG not only acquired unique nuclear localization pattern and ability of recruiting both RXRA and wild-type NUP98, but also exhibited similar transcriptional properties as RARA fusions found in acute promyelocytic leukemia (APL). Using murine bone marrow retroviral transduction/transformation assay, we further demonstrated that NUP98-RARG fusion protein had gained transformation ability of primary hematopoietic stem/progenitor cells, which was critically dependent on the C-terminal GLFG domain of NUP98 and the DNA binding domain (DBD) of RARG. In contrast to other NUP98 fusions, cells transformed by the NUP98-RARG fusion were extremely sensitive to all-trans retinoic acid (ATRA) treatment. Interestingly, while pan-RXR agonists, SR11237 and LGD1069 could specifically inhibit NUP98-RARG transformed cells, mutation of the RXR interaction domain in NUP98-RARG had little effect on its transformation, revealing that therapeutic functions of rexinoid can be independent of the direct biochemical interaction between RXR and the fusion. Together, these results indicate that deregulation of the retinoid/rexinoid signaling pathway has a major role and may represent a potential therapeutic target for NUP98-RARG-mediated transformation.


How research data deliver non-academic impacts: A secondary analysis of UK Research Excellence Framework impact case studies.
This study investigates how research data contributes to non-academic impacts using a secondary analysis of high-scoring impact case studies from the UK's Research Excellence Framework (REF). A content analysis was conducted to identify patterns, linking research data and impact. The most prevalent type of research data-driven impact related to "practice" (45%), which included changing how professionals operate, changing organizational culture and improving workplace productivity or outcomes. The second most common category was "government impacts", including reducing government service costs and enhancing government effectiveness or efficiency. Impacts from research data were developed most frequently through "improved institutional processes or methods" (40%) and developing impact via pre-analyzed or curated information in reports (32%), followed by "analytic software or methods" (26%). The analysis found that research data on their own rarely generate impacts. Instead they require analysis, curation, product development or other forms of significant intervention to leverage broader non-academic impacts.


[Intravenous ultrasonography as a method for imaging the morphoanatomical effects of coronary angioplasty].
Despite the therapeutic success of percutaneous transluminal coronary angioplasty (PTCA), the mechanisms by which PTCA increase vessel luminal size remain uncertain. To better understand the transmural morphologic changes associated with PTCA of stenosed coronary arteries, we studied with a high-frequency intravascular ultrasound catheter 18 coronary artery segments in 18 patients following balloon angioplasty. High-quality cross-sectional images were obtained from 15 coronary sites without complications in all patients. Two distinctive morphologic features following balloon angioplasty were appreciated by intravascular ultrasound imaging. The first pattern, observed in 10 cases (67%), consisted of a stretched plaque without any evidence of dissection. The second pattern, found in 5 cases (33%), demonstrated a dissection of the plaque ranging from a radial tear with separation of the 2 ends of the plaque to an extensive dissection which, in 1 case, encompassed the entire circumference of the artery. Although angiography showed a good post-angioplasty result in all cases, intravascular ultrasound evidenced a large amount of residual atheroma occupying the artery cross-sectional area. In addition, this imaging modality revealed more often than angiography the presence of calcification and dissection. These data demonstrate that PTCA creates different morphologic patterns which are related to the mechanisms of lumen enlargement and that the coronary artery anatomy after dilatation is much more complex than that observed with angiography. This study confirms that intravascular ultrasound is a feasible and safe imaging modality which provides new valuable insight into the mechanisms by which angioplasty improves vessel patency.


Effects of induced aestivation in Protopterus annectens: a histomorphological study.
The aim of this research was to induce, at will in the laboratory, aestivation of the Dipnoan Protopterus annectens, in order to compare the structure of organs in lungfish adapted to aquatic or aestivating conditions. The animals were placed in a glass tank containing warm water, and the bottom of the tank was filled with clay and sand. To start aestivation the water was allowed to slowly evaporate; as soon as the fish was in a dry environment, it began to excavate a hole in the mud and to burrow into it. Scanning electron microscopy and histological techniques compared the morphology of skin, gills, and lungs in aestivating and free-swimming animals. In the aestivating animals, the secondary lamellae of the gills became thick and were covered by mucus that pasted the lamellae together. The epidermis of the skin was thin and composed of layers of flattened cells. In contrast, in free-swimming animals, the secondary lamellae of the gills were widely separated and the epidermis of the skin was thick and contained numerous mucus-laden cells. The lungs, thin bloodless threads in the aquatic conditions, were, in the air-breathing animals, rich in blood and showed thick walls with ridges and pillars that protruded into the lung cavity, producing small alveolar protrusions. The features of the skin and lungs were similar to that of amphibians, testifying to the convergence of some tissue morphology in aquatic animals utilizing land as a cohabitat.


Killing individual poultry on-farm-a survey among veterinarians and farmers.
To date, information about reasons to select and kill poultry on-farm and which method veterinarians and poultry producers preferably use is scarce. Little is also known about their knowledge of the legislation regarding on-farm killing methods and of methods alternative to the one(s) they use, as well as their perception of those alternatives. In this study, Flemish poultry veterinarians (n = 13), broiler chicken producers (n = 27), and turkey producers (n = 4) were surveyed on killing methods they currently use in practice and alternative methods, on their opinion about what constitutes an appropriate method for on-farm killing of poultry, and on their reasons for killing. All poultry veterinarians and chicken producers who filled out the survey kill poultry by manual cervical dislocation (CD), whereas some turkey farmers also indicated killing by percussive blow to the head (n = 1) or exsanguination (n = 1). Turkey producers seem to be more inclined not to kill animals with injuries or symptoms of disease as compared to veterinarians or chicken producers, such that moribund turkeys are more likely to remain in the flock. Veterinarians considered the following properties of a killing method important: animal friendliness, applicability inside the stable, a high success rate, and time efficiency. Producers ranked the properties similarly, but for them, ease of performance and cost-efficiency were more important than applicability inside the stable. Producers scored those killing properties rather positively for manual as well as mechanical CD. Veterinarians and chicken producers considered the captive bolt method to be easy to perform, to have a high success rate, to be feasible to perform in the stable, and to be animal-friendly. Turkey producers, however, had doubts about the latter 2 properties. Gas, injection, and electrocution were inferior methods to kill poultry according to producers. In conclusion, manual CD is the most common method for killing broiler chickens and turkeys, and knowledge of, and experience with, alternative methods is very limited, both among veterinarians and producers. Informing them about legislation and training for the use of alternative killing techniques are recommended.


Progressive supranuclear palsy affects both the substantia nigra pars compacta and reticulata.
We have analyzed the neuropathology of the substantia nigra in four cases of progressive supranuclear palsy compared with age-matched controls and patients with Parkinson's disease. Although there are many reports of severe dopaminergic cell loss in progressive supranuclear palsy, the fate of the GABAergic pars reticulata neurones remains unclear. Serial section analysis and fractional counts of pars compacta neurones (identified by their neuromelanin pigment) and pars reticulata neurones (identified using parvalbumin immunohistochemistry) were performed, and the type and distribution of neuropathology were described. Severe neurodegeneration within the dopaminergic pars compacta was seen in all cases of progressive supranuclear palsy and all cases of Parkinson's disease compared with controls. Lewy body pathology was found only in cases of Parkinson's disease, while neurofibrillary tangles were seen only in cases of progressive supranuclear palsy. Tau-positive astrocytes and neuropil threads were occasionally seen in controls and cases of Parkinson's disease (particularly those of advanced age) but were extremely numerous in all cases of progressive supranuclear palsy. There was a similar decrease in parvalbumin immunoreactivity within the pars reticulata in both progressive supranuclear palsy and Parkinson's disease. However, there was a striking 70% reduction in the number of pars reticulata neurones in progressive supranuclear palsy, with no cell loss observed in Parkinson's disease compared with controls. Our results show that both the dopaminergic pars compacta and the GABAergic pars reticulata are significantly damaged in cases of progressive supranuclear palsy. The distribution of neurodegeneration in patients with Parkinson's disease and progressive supranuclear palsy is discussed with respect to the current theories on pathophysiology in basal ganglia circuitry.


Velopharyngeal incompetence as the presenting symptom of malignant brainstem tumor.
When a patient presents with velopharyngeal incompetence (VPI) without an obvious structural or neurologic cause, the clinician is faced with a diagnostic challenge. We present an 11-year-old male with a long history of VPI who had been referred to our institution for evaluation and treatment. Detailed clinical examination and work-up revealed a malignant brainstem tumor. The presenting symptoms of breathiness associated with VPI had been overlooked by several different clinicians in the past. The patient successfully underwent a sphincter pharyngoplasty. A careful neurologic examination with special attention to the cranial nerves is necessary to identify subtle neurologic deficits and avoid delay in diagnosis. Differential diagnosis of neurogenic VPI is discussed.


Tomoregulin (TMEFF2) Binds Alzheimer's Disease Amyloid-β (Aβ) Oligomer and AβPP and Protects Neurons from Aβ-Induced Toxicity.
Amyloid-β (Aβ) protein causes neurotoxicity and its abnormal aggregation into amyloid is a pathological hallmark of Alzheimer's disease (AD). Cellular proteins able to interact with Aβ or its precursor, AβPP (amyloid-β protein precursor), may regulate Aβ production and neurotoxicity. We identified a brain-enriched type I transmembrane protein, tomoregulin (TR), that directly binds Aβ and Aβ oligomers (AβO). TR co-immunoprecipitated with Aβ and AβO in cultured cells and co-localized with amyloid plaques and intraneuronal Aβ in the 5xFAD AD mouse model. TR was also enriched in astrocytic processes reactive to amyloid plaques. Surface plasmon resonance spectroscopy studies showed that the extracellular domain of TR binds to AβO with a high affinity (KD = 76.8 nM). Electron paramagnetic resonance spectroscopy also demonstrated a physical interaction between spin-labeled Aβ and the TR extracellular domain in solution. Furthermore, TR also interacted with AβPP and enhanced its cleavage by α-secretase. Both cellular expression of TR and application of recombinant TR extracellular domain protected N2a neurons from AβO-induced neuronal death. These data provide first evidence that neuronal and astrocytic expression of TR is intimately related to Aβ metabolism and toxicity, and could be neuroprotective through its direct interaction with Aβ and AβPP.


Dietary behaviors mediate the association between food insecurity and obesity among socioeconomically disadvantaged youth.
Across many Asian countries, the prevalence of obesity is increasing and is increasingly recognized as an emerging public health issue. Food insecurity, defined as the limited or uncertain availability of nutritionally adequate and safe foods, is major difficulty faced by many poor subpopulations despite national economic growth. Previous research has yielded inconsistent results about the association between food insecurity and obesity, with positive associations more consistently observed among women than men. This study aims to elucidate relationships between food insecurity, dietary behaviors, and weight status, stratified by gender. Data come from the Taiwan Database of Children and Youth in Poverty. Participants (n = 1326) aged 10-18 years (2009-2013), which included measures of food security and dietary behaviors in three survey waves. Weight status was determined using self-reported body mass index (BMI; kg/m2) with the category "overweight/obesity" defined as a BMI above the 85th percentile within a representative population. Logistic regression analyses indicated a significant association between food insecurity and obesity/overweight with an odds ratio (OR) = 1.45, 95% confidence interval (CI) 1.25-1.70. Food insecurity was associated with a select number of unhealthy dietary behaviors. Skipping breakfast in females (OR = 1.63, 95% CI 1.20-2.22) and consuming snacks and sugared drinks in males (OR = 1.51, 95% CI 1.15-1.98) were associated with obesity/overweight. Food insecurity remained significantly associated with obesity/overweight after adjusting for dietary behaviors. In addition, there was some evidence that dietary behaviors mediated the association between food security and obesity/overweight. In conclusion, economically disadvantaged youth are vulnerable to food insecurity and obesity, and food insecurity may increase the likelihood of obesity due to its association with dietary behaviors. Understanding the mediating factors between food insecurity and obesity may help to amend weight-related health problems in poverty-affected populations.


Mapping the hemagglutination domain of rotaviruses.
Most strains of animal rotaviruses are able to agglutinate erythrocytes, and the surface protein VP4 is the virus hemagglutinin. To map the hemagglutination domain on VP4 while preserving the conformation of the protein, we constructed full-length chimeras between the VP4 genes of hemagglutinating (YM) and nonhemagglutinating (KU) rotavirus strains. The parental and chimeric genes were expressed in insect cells, and the recombinant VP4 proteins were evaluated for their capacity to agglutinate human type O erythrocytes. Three chimeric genes, encoding amino acids 1 to 208 (QKU), 93 to 208 (QC), and 93 to 776 (QYM) of the YM VP4 protein in a KU VP4 background, were constructed. YM VP4 and chimeras QKU and QC were shown to specifically hemagglutinate, indicating that the region between amino acids 93 and 208 of YM VP4 is sufficient to determine the hemagglutination activity of the protein.


The Myopic Shift in Aphakic Eyes in the Infant Aphakia Treatment Study After 10 Years of Follow-up.
To report the myopic shift in the aphakic eyes of a cohort of children who underwent unilateral cataract surgery during infancy and were then followed longitudinally for 10.5 years. One-half of the children enrolled in the Infant Aphakia Treatment Study (IATS) were randomized to aphakia and contact lens correction after unilateral cataract surgery. They then underwent ocular examinations using standardized protocols at prescribed time intervals until age 10.5 years. Thirty of 57 children randomized to aphakia remained aphakic at age 10.5, having undergone unilateral cataract surgery at a median age of 1.6 (IQR: 1.1-3.1) months. The median refractive error (RE) in the 57 eyes randomized to aphakia immediately after cataract surgery was 19.01 D (IQR: 16.98-20.49) compared to 10.38 D (IQR: 7.50-14.00) for the 30 eyes that remained aphakic at age 10.5 years. The mean change in RE in aphakic eyes was -2.11 D/year up to age 1.5 years, -0.68 D/year from 1.5 to 5.0 years, and -0.35 D/year from age 5 to 10.5 years. At age 10.5 years, 18 patients continued to wear a contact lens correction (silicone elastomer, n=6; gas permeable, n=6; hydrogel, n=5; and silicone hydrogel, n=1) (median RE, 12.50 D), 9 wore only spectacles (median RE, 4.00 D), and 4 wore no correction (median RE, 11.25 D) to correct their aphakic eye. The RE in aphakic eyes decreased by 44% from infancy to age 10.5 years. About two-thirds of children who remained aphakic at age 10.5 years continued to wear a contact lens.


Different modifications of phosphorylated Smad3C and Smad3L through TGF-β after spinal cord injury in mice.
Transforming growth factor-β (TGF-β) is an anti-inflammatory cytokine and is expressed in the injured spinal cord. TGF-β signals through receptors to activate Smad proteins, which translocate into the nucleus. In the present study, we investigated the chronological alterations and cellular locations of the TGF-β/Smad signaling pathway following spinal cord injury (SCI) in mice. ELISA analysis showed that the concentration of interleukin-6 (IL-6) in injured spinal cords significantly increases immediately after SCI, while the concentration of TGF-β gradually increased after SCI, peaked at 2 days, and then gradually decreased. Immunohistochemical studies revealed that Smad3 was mainly expressed in neurons of the spinal cord. Phosphorylated Smad3 at the C-terminus (p-Smad3C) was stained within the motor neurons in the anterior horn, while phosphorylated Smad3 at the linker regions (p-Smad3L) was expressed in astrocytes within gray matter. These findings suggest that SCI induces gradual increases in TGF-β and induces different activation of p-Smad3C and p-Smad3L. Phosphorylated Smad3C might be involved in neuronal degeneration after SCI, and p-Smad3L may play a role in glial scar formation by astrocytes.


Are Suburban Youth Becoming More Traditional? A Comparative Study on Young People from Muslim Backgrounds Living on the Outskirts of Madrid and Paris.
This paper compares how Muslim youth identify with traditional values in two disadvantaged areas: a banlieue of Paris and a barrio of Madrid. Research has revealed divergent forms of identification with Islam and tradition. In Les Bosquets (Paris), where the predominating context is one of ethnic segregations, a lack of civic participation, isolation from the city center, and increasing inequalities and Islamophobia, youth are exhibiting an ambivalent return to traditional values and building a new proud, combative and collective Islam. In the case of Madrid, young people distinguish between human/religious and traditional values while ignoring the latter values when they interfere with individual interests. Islam here is individual and compatible with a collective feeling of neighborhood, one built within a context of ethnic diversity, a less intense feeling of Islamophobia, greater accessibility to the cosmopolitan urban center, and dense networks of civil society that encourage local participation.


Requirements for the self-directed replication of flock house virus RNA 1.
The larger segment (RNA 1) of the bipartite, positive-sense RNA genome of the nodavirus flock house virus encodes the viral RNA-dependent RNA polymerase. Two nonstructural viral proteins are made during the self-directed replication of this RNA: protein A (110 kDa), the translation product of RNA 1 itself, and protein B (11 kDa), the translation product of a subgenomic RNA (RNA 3) that is produced from RNA 1 during replication. To examine the roles of these proteins in RNA replication, specialized T7 transcription plasmids that contained wild-type or mutant copies of flock house virus RNA 1 cDNA were constructed and used in cells infected with the vaccinia virus-T7 RNA polymerase recombinant to make full-length transcripts that directed their own replication. Sequences in the primary transcripts that extended beyond the ends of the authentic RNA 1 sequence inhibited self-directed RNA replication, but plasmids that were constructed to minimize these terminal extensions produced primary transcripts that replicated as abundantly as authentic RNA 1. Truncation or mutation of the open reading frame for protein A eliminated self-directed replication, although the mutant RNA 1 remained a competent template for replication by wild-type protein A supplied in trans. These results showed that protein A was essential for RNA replication and that the process was not inseparably coupled to complete translation of the template. In contrast, protein B could be eliminated without inhibiting replication by mutations that disrupted the second of the two overlapping open reading frames on RNA 3. Furthermore, a mutant of RNA 1 in which the first nucleotide of the RNA 3 region was changed from G to U replicated at levels as high as those of the wild type without making either RNA 3 or protein B. However, diminishing replication levels were observed during subsequent replicative passages of RNA from both the mutants that could not make protein B. Roles for this protein that could account for the subtle phenotype of these mutants are discussed.


Physiology of sedentary behavior and its relationship to health outcomes.
This article reports on the findings and recommendations of the "Physiology of Sedentary Behavior and Its Relationship to Health Outcomes" group, a part of a larger workshop entitled Sedentary Behavior: Identifying Research Priorities sponsored by the National Heart, Lung, and Blood Institute and by the National Institute on Aging, which aimed to establish sedentary behavior research priorities. The discussion within our workshop led to the formation of critical physiological research objectives related to sedentary behaviors, that is, if appropriately researched, would greatly affect our overall understanding of human health and longevity. Primary questions are related to physiological "health outcomes" including the influence of physical activity versus sedentary behavior on the function of a number of critical physiological systems (aerobic capacity, skeletal muscle metabolism and function, telomeres/genetic stability, and cognitive function). The group also derived important recommendations related to the "central and peripheral mechanisms" that govern sedentary behavior and how energy balance has a role in mediating these processes. General recommendations for future sedentary physiology research efforts indicate that studies of sedentary behavior, including that of sitting time only, should focus on the physiological effect of a "lack of human movement" in contradistinction to the effects of physical movement and that new models or strategies for studying sedentary behavior-induced adaptations and links to disease development are needed to elucidate underlying mechanism(s).


Pathology and genomics of pediatric melanoma: A critical reexamination and new insights.
The clinicopathologic features of pediatric melanoma are distinct from those of the adult counterpart. For example, most childhood melanomas exhibit a uniquely favorable biologic behavior, save for those arising in large/giant congenital nevi. Recent studies suggest that the characteristically favorable biologic behavior of childhood melanoma may be related to extreme telomere shortening and dysfunction in the cancer cells. Herein, we review the genomic profiles that have been defined for the different subtypes of pediatric melanoma and particularly emphasize the potential prognostic value of telomerase reverse transcriptase alterations for these tumors.


[Effect of carbidine on parameters of ethanol oxidation in experimental alcoholic intoxication].
The effect of carbidine on enzymes of ethanol and acetaldehyde oxidation, the rate of ethanol elimination and the parameters of ethanol consumption were studied during long-term alcoholic intoxication. Carbidine administration was shown to increase the activity of alcohol dehydrogenase of the liver tissue, to decrease the activity of aldehyde dehydrogenase with a low Km to acetaldehyde. Also, the rate of ethanol elimination and a relative amount of consumed ethanol increase at the expense of an increase of the volume of consumed liquid.


Secondary acute leukaemias with 11q23 rearrangement: clinical, cytogenetic, FISH and FICTION studies.
Three patients with secondary acute leukaemia after treatment with topoisomerase II inhibitor agents are described. Two patients had acute myeloid leukaemia (AML). FAB M5a, one had pro-B-acute lymphoblastic leukaemia (ALL). The interval between initiation of chemotherapy and the onset of secondary acute leukaemia was 19-20 months. 11q23 rearrangements were detected in all cases. They were due to translocations t(11;19) (q23;p13.3), t(11;16)(q23;p13) and t(4;11)(q21;q23), respectively. Fluorescence in situ hybridization (FISH) with Yeast Artificial Chromosome (YAC) probe 13HH4 spanning the ALL-1 gene on 11q23 confirmed that in each case the ALL-1 gene had been disrupted by the translocations. The study underlined the relationship between the development of secondary acute leukaemias with 11q23 rearrangement and previous chemotherapy with topisomerase II inhibitor agents. So far, however, only six adult patients with secondary ALL with t(4;11) after treatment with topoisomerase II inhibitor agents have been reported. All with t(4;11) mostly occurs in infants or young children. Our patient received epirubicin continuously for >19 months. This indicates that both myeloid and lymphoid leukaemias with involvement of the ALL-1 gene can be induced by exogenous agents, especially topoisomerase II inhibitors. Thus they may have a common biological background. This hypothesis was substantiated by means of combined immunophenotyping and FISH (FICTION). In the case of AML M5a with t(11;19), the tumour cells with ALL-1 rearrangement expressed CD34. Moreover, the pro-B-ALL with t(4;11) was CD34 positive. These findings suggest that the cell of origin of secondary AML and ALL with 11q23 rearrangement is an immature haemopoietic progenitor cell.


Androgen regulation of murine beta-glucuronidase expression: identification and characterization of a nonresponse variant.
One of the major features of beta-glucuronidase (GUS) expression in inbred strains of the house mouse, Mus musculus, is the responsiveness of this enzyme to androgen stimulation in tubule cells of the kidney. Both GUS-specific and nonspecific mutations have been described which define genes that serve to control this response. During examination of the expression of GUS in the interbreeding subspecies, Mus hortulanus, a new GUS haplotype was uncovered that is characterized, in part, by a lack of GUS response to androgen stimulation in an apparently responsive kidney. Blot hybridization analyses of kidney RNA with a radiolabeled murine GUS cDNA shows this lack of response to be reflected in GUS mRNA levels. The difference in heat stability of GUS activity between M. hortulanus and a responsive inbred strain, ICR/Ha, was utilized to assess the contribution of each parent to kidney levels of GUS in androgen-treated and -untreated F1 progeny of these strains. The results, together with preliminary genetic studies, suggest that the element controlling this responsiveness (or the lack thereof) is cis-active and tightly linked to the GUS structural gene on chromosome 5. It is not known whether this element is identical to another GUS-specific, cis-active element, Gus-r, which also controls the androgen response of GUS in mouse kidney.


Effects of tobacco genetically modified to express protease inhibitor bovine spleen trypsin inhibitor on non-target soil organisms.
Effects of tobacco genetically modified to express the protease inhibitor bovine spleen trypsin inhibitor (BSTI) were examined in laboratory assays against three earthworm and one collembolan species. BSTI is a serine protease inhibitor that can bind to the digestive trypsins of insects feeding on modified plants, resulting in reduced growth and survival. Protease inhibitors are active against a broad range of insects, so may have a large impact on non-target organisms. Survival and fecundity of the collembolan Folsomia candida were unaffected by consumption of artificial diet containing BSTI-expressing tobacco leaf or powdered freeze-dried BSTI-expressing tobacco leaf that was added to soil. Similarly, mortality and growth of earthworms Aporrectodea caliginosa and Lumbricus rubellus did not differ significantly between soil augmented with BSTI-expressing tobacco leaves or unmodified control leaves. The redworm Eisenia fetida gained less weight when provided with BSTI-expressing leaves in one assay, but when the experiment was repeated, there was no significant difference between treatments. BSTI-expressing tobacco and unmodified control leaves decomposed at the same rate, indicating that the inhibitor had no effect on the overall function of the decomposer community of micro-flora and fauna in soil.


Theoretical and kinetic study of the reactions of ketones with HO2 radicals. Part I: abstraction reaction channels.
This work presents an ab initio and chemical kinetic study of the reaction mechanisms of hydrogen atom abstraction by the HO2 radical on five ketones: dimethyl, ethyl methyl, n-propyl methyl, iso-propyl methyl, and iso-butyl methyl ketones. The Møller-Plesset method using the 6-311G(d,p) basis set has been used in the geometry optimization and the frequency calculation for all the species involved in the reactions, as well as the hindrance potential description for reactants and transition states. Intrinsic reaction coordinate calculations were carried out to validate all the connections between transition states and local minima. Energies are reported at the CCSD(T)/cc-pVTZ//MP2/6-311G(d,p) level of theory. The CCSD(T)/cc-pVXZ method (X = D, T, Q) was used for the reaction mechanism of dimethyl ketone + HO2 radical in order to benchmark the computationally less expensive method of CCSD(T)/cc-pVTZ//MP2/6-311G(d,p). High-pressure limit rate constants have been calculated for all the reaction channels by conventional transition state theory with asymmetric Eckart tunneling corrections and 1-D hindered rotor approximations in the temperature range 500-2000 K.


Effect of macrophage colony-stimulating factor on anticryptococcal activity of bronchoalveolar macrophages: synergy with fluconazole for killing.
The anticryptococcal activity of murine bronchoalveolar macrophages (BAM) and their synergy with fluconazole (FCZ) was studied. BAM cultured with tissue culture medium for 48 to 72 h were fungicidal (24 to 39%) in a 3-h killing assay. However, net killing of Cryptococcus neoformans did not continue when culture time was extended to 24 h, although BAM were fungistatic (88 to 98%). Treatment with macrophage colony-stimulating factor (M-CSF; 5,000 U/ml, 48 h) did not significantly increase BAM killing of a low challenge dose in 3-h assays compared with control BAM. However, M-CSF-treated BAM were significantly more fungistatic against higher challenge doses in the 3-h assays. FCZ was not fungicidal at 5 micrograms/ml but was highly fungistatic (98 and 99% at 24 and 48 h, respectively). M-CSF-treated BAM acted synergistically with FCZ (2.5 micrograms/ml) for significantly greater killing than control BAM, 55% versus 20% and 96% versus 45% at 24 h and 48 h, respectively. Killing by M-CSF BAM and FCZ (5.0 micrograms/ml) was significantly (P < 0.01) greater than that by control BAM and FCZ at 48 h. These findings indicate an important collaborative role for BAM and FCZ in killing C. neoformans, and this is enhanced by M-CSF.


Stress and angiotensin II: novel therapeutic opportunities.
Angiotensin II was initially described as a hormone of peripheral origin, the active end product of the Renin-Angiotensin System. The subsequent discovery that Angiotensin II was locally formed and selectively regulated in most organs indicated that tissue Angiotensin II systems might play additional important roles. After initial controversy, the presence of an Angiotensin II system in the brain is now universally accepted. Brain Angiotensin II is probably involved in the regulation of many brain functions. Angiotensin II AT1 receptors are localized not only in areas related to the regulation of autonomic and endocrine control, but also in many other areas of the brain involved in emotional, sensory and motor functions. Angiotensin II AT2 receptors are more abundant in brain areas related to sensory and motor control. The roles of brain Angiotensin II appear to be multiple and complex. In addition to a regulatory role in the control of the autonomic and hormone systems, the peptide participates in brain development, sensory processes, cognition and in the regulation of cerebrovascular flow. Recent developments indicate that blockade of the brain Angiotensin II AT1 receptors not only contributes to a significant blood pressure decrease in hypertension, but that simultaneous antagonism of peripheral and brain AT1 receptors reduces the sympathoadrenal and hormonal responses to stress and prevents stress-induced gastric injury. A novel role emerges for the use of peripheral and centrally acting AT1 receptor antagonists as therapeutically advantageous for the treatment of stress-related disorders.


The miR-182-5p/FGF21/acetylcholine axis mediates the crosstalk between adipocytes and macrophages to promote beige fat thermogenesis.
A dynamically regulated microenvironment, which is mediated by crosstalk between adipocytes and neighboring cells, is critical for adipose tissue homeostasis and function. However, information on key molecules and/or signaling pathways regulating the crosstalk remains limited. In this study, we identify adipocyte miRNA-182-5p (miR-182-5p) as a crucial antiobesity molecule that stimulated beige fat thermogenesis by promoting the crosstalk between adipocytes and macrophages. miR-182-5p was highly enriched in thermogenic adipocytes, and its expression was markedly stimulated by cold exposure in mice. In contrast, miR-182-5p expression was significantly reduced in adipose tissues of obese humans and mice. Knockout of miR-185-5p decreased cold-induced beige fat thermogenesis whereas overexpression of miR-185-5p increased beiging and thermogenesis in mice. Mechanistically, miR-182-5p promoted FGF21 expression and secretion in adipocytes by suppressing nuclear receptor subfamily 1 group D member 1 (Nr1d1) at 5'-UTR, which in turn stimulates acetylcholine synthesis and release in macrophages. Increased acetylcholine expression activated the nicotine acetylcholine receptor in adipocytes, which stimulated PKA signaling and consequent thermogenic gene expression. Our study reveals a key role of the miR-182-5p/FGF21/acetylcholine/acetylcholine receptor axis that mediates the crosstalk between adipocytes and macrophages to promote beige fat thermogenesis. Activation of the miR-182-5p-induced signaling pathway in adipose tissue may be an effective approach to ameliorate obesity and associated metabolic diseases.


Acinetobacter gandensis sp. nov. isolated from horse and cattle.
We previously reported the presence of an OXA-23 carbapenemase in an undescribed species of the genus Acinetobacter isolated from horse dung at the Faculty of Veterinary Medicine, Ghent University, Belgium. Here we include six strains to corroborate the delineation of this taxon by phenotypic characterization, DNA-DNA hybridization, 16S rRNA gene and rpoB sequence analysis, % G+C determination, MALDI-TOF MS and fatty acid analysis. The nearly complete 16S rRNA gene sequence of strain UG 60467(T) showed the highest similarities with those of the type strains of Acinetobacter bouvetii (98.4 %), Acinetobacter haemolyticus (97.7 %), and Acinetobacter schindleri (97.2 %). The partial rpoB sequence of strain UG 60467(T) showed the highest similarities with 'Acinetobacter bohemicus' ANC 3994 (88.6 %), A. bouvetii NIPH 2281 (88.6 %) and A. schindleri CIP 107287T (87.3 %). Whole-cell MALDI-TOF MS analyses supported the distinctness of the group at the protein level. The predominant fatty acids of strain UG 60467(T) were C12 : 0 3-OH, C12 : 0, C16 : 0, C18 : 1ω9c and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH). Strains UG 60467(T) and UG 60716 showed a DNA-DNA relatedness of 84 % with each other and a DNA-DNA relatedness with A. schindleri LMG 19576(T) of 17 % and 20 %, respectively. The DNA G+C content of strain UG 60467(T) was 39.6 mol%. The name Acinetobacter gandensis sp. nov. is proposed for the novel taxon. The type strain is UG 60467(T) ( = ANC 4275(T) = LMG 27960(T) = DSM 28097(T)).


The behavior of thoracic trabecular bone during flexion.
Thoracic compression fractures are often described as anterior wedge fractures. Although the radiographic signs of these fractures are easily identified, the mechanism of the trabecular failure is not well understood. The current study addressed this mechanism in the lower thoracic spine by measuring the trabecular strain. Trabecular strain was measured in six human thoracic cadaver spines during 1) compressive and 2) flexural loading. The strains were measured at incremental loads using a texture correlation. They were analyzed by global contour plots and regional analysis of the T11 vertebrae. Specimens loaded under only compression exhibited uniform strains in the vertebral body. During flexion, however, the strains were concentrated in the anterosuperior margin of the vertebral body and the compressive and shear strain magnitudes in this region were significantly increased. These results demonstrate that the flexural position places the lower thoracic spine at greater risk of anterior compression fracture as seen clinically.


Reduction of iron and synthesis of protoheme by Spirillum itersonii and other organisms.
Membranes from Spirillum itersonii reduce ferric iron to ferrous iron with reduced nicotinamide adenine dinucleotide or succinate as a source of reductant. Iron reduction was measured spectrophotometrically at 562 nm using ferrozine, which chelates ferrous iron specifically. Reduced nicotinamide adenine dinucleotide or succinate was also effective as a source of iron. The effects of respiratory inhibitors suggested that reduction of iron occurs at one or more sites on the respiratory chain before cytochrome c. Reduction of iron and synthesis of protoheme with the physiological reductants were also observed with crude extracts of other bacteria, including Rhodopseudomonas spheroides, Rhodopseudomonas capsulata, Paracoccus denitrificans, and Escherichia coli. The effect of oxygen upon reduction of iron and formation of protoheme was examined with membranes from S. itersonii, using succinate as a source of reductant. Both systems were inhibited by oxygen, but this effect was completely reversed by addition of antimycin A. We conclude that reduced components of the respiratory chain serve as reductants for ferric iron, but with oxygen present they are oxidized preferentially by the successive members of the chain. This could be a mechanism for regulating synthesis of heme and cytochrome by oxygen.


Reliable microfluidic on-chip incubation of droplets in delay-lines.
Together with droplet creation, fusion and sorting, the incubation of droplets is one of the most important and essential operations for droplet-based microfluidic assays. This manuscript concerns the development of delay-lines, which are necessary to allow incubation of reactions for precise time periods. We analyze the problems associated with creating delay-lines for incubation in the minute to hour time range, which arise from back-pressure and from the dispersion in the incubation time due to the unequal speeds with which droplets pass through the delay-line. We describe delay-line systems which resolve these problems and demonstrate their use to measure reaction kinetics over several minutes in droplets.


Clinical presentation of childhood neuroborreliosis; neurological examination may be normal.
Neuroborreliosis has its highest incidence in children and the older people. Signs and symptoms are different between the different age groups. The aim of this study was to describe the clinical spectrum of neuroborreliosis in children. The Dutch Paediatric Surveillance system registered cases of childhood neuroborreliosis during 2 years. All Dutch paediatric hospitals took part in this surveillance. Criteria for reporting cases were strictly defined. 89 cases of neuroborreliosis were reported; in 66 cases diagnosis was confirmed. Facial weakness was one of the presenting symptoms in 47 cases (71%) and the only symptom in nine children (14%). The five complaints most frequently reported were: malaise, headache, fatigue, fever and neck pain. 52 children (79%) had one or more objective neurological signs at presentation, of which facial nerve palsy, other cranial nerve abnormalities and meningeal signs were most frequent. 14 patients (21%), however, had no neurological signs at physical examination. In these patients, the number of subjective complaints was higher, and the time interval to diagnosis was longer compared with those with objective neurological abnormalities. In this study, 79% of paediatric neuroborreliosis patients presented with neurological signs, most often facial nerve palsy. 21% presented in an atypical way without neurological signs. A thorough neurological examination is essential once neuroborreliosis is considered in children. Even in the absence of neurological signs, neuroborreliosis may be suspected in children with typical antecedents and multiple symptoms. Cerebrospinal fluid investigations are then required to confirm the diagnosis.


Clinical assessment of balance using BBS and SARAbal in cerebellar ataxia: Synthesis of findings of a psychometric property analysis.
In the previous psychometric analysis paper in our series for identifying the core set of balance measures for the assessment of balance, we recommended the Berg Balance Scale (BBS) and balance sub-components of the Scale for the assessment and rating of ataxia (SARAbal) as psychometrically sound measures of balance for people with cerebellar ataxia (CA) secondary to multiple sclerosis. The present study further examined the suitability of BBS and SARAbal for the assessment of balance in CA with regard to psychometric property strength, appropriateness, interpretability, precision, acceptability and feasibility. Criteria to fulfill each factor was defined according to the framework of Fitzpatrick et al. (1998). Based on the findings of our previous psychometric analysis, each criterion was further analyzed. The psychometric analysis reported good reliability and validity estimates for the BBS and SARAbal recommending them as psychometrically sound measures; they fulfilled both criteria for appropriateness and interpretability, the measures showed evidence for precision and acceptability, and they were found to be feasible in terms of the time and cost involved for the balance assessment. We have provided evidence for the use of the BBS and SARAbal for the assessment of balance among people with CA.


Exercise reduces appetite and traffics excess nutrients away from energetically efficient pathways of lipid deposition during the early stages of weight regain.
The impact of regular exercise on energy balance, fuel utilization, and nutrient availability, during weight regain was studied in obese rats, which had lost 17% of their weight by a calorie-restricted, low-fat diet. Weight reduced rats were maintained for 6 wk with and without regular treadmill exercise (1 h/day, 6 days/wk, 15 m/min). In vivo tracers and indirect calorimetry were then used in combination to examine nutrient metabolism during weight maintenance (in energy balance) and during the first day of relapse when allowed to eat ad libitum (relapse). An additional group of relapsing, sedentary rats were provided just enough calories to create the same positive energy imbalance as the relapsing, exercised rats. Exercise attenuated the energy imbalance by 50%, reducing appetite and increasing energy requirements. Expenditure increased beyond the energetic cost of the exercise bout, as exercised rats expended more energy to store the same nutrient excess in sedentary rats with the matched energy imbalance. Compared with sedentary rats with the same energy imbalance, exercised rats exhibited the trafficking of dietary fat toward oxidation and away from storage in adipose tissue, as well as a higher net retention of fuel via de novo lipogenesis in adipose tissue. These metabolic changes in relapse were preceded by an increase in the skeletal muscle expression of genes involved in lipid uptake, mobilization, and oxidation. Our observations reveal a favorable shift in fuel utilization with regular exercise that increases the energetic cost of storing excess nutrients during relapse and alterations in circulating nutrients that may affect appetite. The attenuation of the biological drive to regain weight, involving both central and peripheral aspects of energy homeostasis, may explain, in part, the utility of regular exercise in preventing weight regain after weight loss.