Comparison of bidisomide, flecainide and dofetilide on action potential duration in isolated canine atria: effect of isoproterenol.
Prolongation of action potential duration (APD) and the effective refractory period (ERP) is one mechanism to prevent reentrant atrial and ventricular arrhythmias. Because arrhythmias are usually associated with an elevated sympathetic tone and increase of circulating catecholamines, the potential influence of catecholamines on antiarrhythmic effects of an agent are critical to predicting the potential clinical response. In this study the effects of three different antiarrhythmic agents, bidisomide, flecainide and dofetillide, each of which prolongs atrial ERP, were compared before and after treatment with isoproterenol, a beta-adrenergic agonist. Standard intracellular microelectrode recording techniques were used to record action potentials from isolated canine atrial tissue. Bidisomide and flecainide elicited a 20 to 27 msec increase in APD and ERP that was independent of stimulation frequency (1-5 Hz). Dofetilide prolonged APD and ERP at 1 Hz (40 and 37 msec, respectively) but was completely ineffective at 5 Hz. After equilibration with the anti-arrhythmic agent, tissues were additionally exposed to isoproterenol. Only bidisomide prolonged APD and ERP in the presence of isoproterenol. In the converse series of experiments after treatment with isoproterenol, which caused a 20 to 30% reduction in APD and ERP, only bidisomide completely reversed the effect of 1 microM isoproterenol. Bidisomide was inactive in a functional beta-adrenergic antagonism assay, thus ruling out beta-adrenergic blockade as a potential mechanism. These results indicate that bidisomide, unlike flecainide and dofetilide was able to prolong APD and ERP in isolated canine atrium even at high stimulation frequencies and in the presence of isoproterenol. These data suggest that bidisomide would be effective in the presence of elevated sympathetic tone and on the agents studied, only bidisomide possessed a unique and desirable antiarrhythmic profile.


Hyperglycemia limits experimental aortic aneurysm progression.
Diabetes mellitus (DM) is associated with reduced progression of abdominal aortic aneurysm (AAA) disease. Mechanisms responsible for this negative association remain unknown. We created AAAs in hyperglycemic mice to examine the influence of serum glucose concentration on experimental aneurysm progression. Aortic aneurysms were induced in hyperglycemic (DM) and normoglycemic models by using intra-aortic porcine pancreatic elastase (PPE) infusion in C57BL/6 mice or by systemic infusion of angiotensin II (ANG) in apolipoprotein E-deficient (ApoE(-/-)) mice, respectively. In an additional DM cohort, insulin therapy was initiated after aneurysm induction. Aneurysmal aortic enlargement progression was monitored with serial transabdominal ultrasound measurements. At sacrifice, AAA cellularity and proteolytic activity were evaluated by immunohistochemistry and substrate zymography, respectively. Influences of serum glucose levels on macrophage migration were examined in separate models of thioglycollate-induced murine peritonitis. At 14 days after PPE infusion, AAA enlargement in hyperglycemic mice (serum glucose ≥ 300 mg/dL) was less than that in euglycemic mice (PPE-DM: 54% ± 19% vs PPE: 84% ± 24%, P < .0001). PPE-DM mice also demonstrated reduced aortic mural macrophage infiltration (145 ± 87 vs 253 ± 119 cells/cross-sectional area, P = .0325), elastolysis (% residual elastin: 20% ± 7% vs 12% ± 6%, P = .0209), and neovascularization (12 ± 8 vs 20 ± 6 vessels/high powered field, P = .0229) compared with PPE mice. Hyperglycemia limited AAA enlargement after ANG infusion in ApoE(-/-) mice (ANG-DM: 38% ± 12% vs ANG: 61% ± 37% at day 28). Peritoneal macrophage production was reduced in response to thioglycollate stimulation in hyperglycemic mice, with limited augmentation noted in response to vascular endothelial growth factor administration. Insulin therapy reduced serum glucose levels and was associated with AAA enlargement rates intermediate between euglycemic and hyperglycemic mice (PPE: 1.21 ± 0.14 mm vs PPE-DM: 1.00 ± 0.04 mm vs PPE-DM + insulin: 1.14 ± 0.05 mm). Hyperglycemia reduces progression of experimental AAA disease; lowering of serum glucose levels with insulin treatment diminishes this protective effect. Identifying mechanisms of hyperglycemic aneurysm inhibition may accelerate development of novel clinical therapies for AAA disease.


Alpha-lipoic acid ameliorates oxidative stress by increasing aldehyde dehydrogenase-2 activity in patients with acute coronary syndrome.
Aldehyde dehydrogenase-2 (ALDH2) is the main enzyme responsible for acetaldehyde oxidation in ethanol metabolism and also provides protection against oxidative stress. Alpha-lipoic acid (α-LA), a natural dithiol compound with antioxidant properties, has been reported to increase ALDH2 activity in cultured cells. We analyzed the therapeutic efficacy of α-LA in 63 patients with confirmed acute coronary syndrome (ACS). These patients (52 men and 11 women, with age range 49-72 years) were randomized into two groups: untreated group (n = 30) and α-LA group (n = 33). Patients in the α-LA group were given an intravenous injection of 600 mg α-LA every day for 5 days while the patients in the untreated group were given saline. An isoprostane, 8-iso-prostaglandin F2α (8-iso-PGF2α), one product of arachidonic acid metabolism, was measured as a marker for oxidative stress. The serum levels of 8-iso-PGF2α and ALDH2 activity were determined at admission to the hospital (time 0), and at 24 hours and 1 week after treatment. At 24 hours and 1 week after treatment, ALDH2 activity was significantly higher in the α-LA group than in the untreated group (P < 0.05), whereas the levels of 8-iso-PGF2α were significantly lower in the α-LA group than in the untreated group (all P < 0.05). Importantly, the decrease of 8-iso-PGF2α levels correlated with the increased ALDH2 activity at both 24 hours (r = 0.6234, P < 0.001) and 1 week after treatment (r = -0.3941, P = 0.0014). α-LA may ameliorate oxidative stress through up-regulating ALDH2 activity in patients with ACS.


Endemicity of spotted fever group rickettsiae in Connecticut.
To compare rickettsial infectivity and seropositivity rates against spotted fever group (SFG) rickettsiae, ticks and wild mammals were collected from three areas where Rickettsia rickettsii was thought to be enzootic in Connecticut during 1978-1979, and from four additional sites (with no reported human cases) between 1976 and 1979. Of the 1,001 Dermacentor variabilis examined by the hemolymph test, 59 (5.9%) contained rickettsia-like organisms; direct immunofluorescence tests verified the presence of SFT rickettsiae in 14 specimens. Prevalence of rickettsiae-infected ticks at Newtown, an area where human cases of Rocky Mountain spotted fever probably originated, was 2.2%. Rates for six other areas ranged between 0 and 6.3%. Isolations included Rickettsia montana from four ticks collected at Branford and Woodbridge, and R. rickettsii (R-like strain) from the blood of an acutely ill person. Microagglutination (MA) tests indicated that 15 (14.9%) of 101 Peromyscus leucopus (white-footed mice) from Newtown had agglutinins in titers greater than or equal to 1:8 against R. rickettsii, whereas five of 92 white-footed mice (5.4%) from Brandord, West Hartford, Woodbridge, and Sharon were considered MA-positive. Indirect microimmunofluorescence tests of Procyon lotor (raccoon) sera revealed antibodies to R. rickettsii in 33 of 69 (47.8%) samples from Newtown and in two of 60 (3.3%) from Guilford. Additionally, 17 raccoons had sera specific to R. montana (n = 8) or to the 369-C rickettsia strain (n = 9). Since rickettsia-positive ticks are high-titered seropositive mammals occurred at widely separated sites in Connecticut there are probably several foci of SFG rickettsiae distributed throughout the D. variabilis range.


Circulating miRNAs and their target genes associated with arsenism caused by coal-burning.
Endemic arsenism, caused by burning coal containing high levels of arsenic, is found only in the Guizhou and Shanxi Provinces of China. Dysregulated microRNAs (miRNAs), detected in the blood, are emerging as promising biomarkers. At present, little is known about the change and clinical efficacy of circulating miRNAs in patients with endemic arsenism produced by burning of coal. Here, we determined, by using TaqMan Human miRNA Array Chips, the differential expression of plasma miRNAs between patients with arsenism caused by coal-burning and a control group. Four increased miRNAs (miR-21, miR-145, miR-155, and miR-191) were verified in a larger sample by quantitative real-time PCR. Furthermore, bioinformatics and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were used to associate changes in plasma levels of the miRNAs with their functions and their effects on various pathways. The results of chip array assays show that the levels of miR-21, miR-141, miR-148a, miR-145, miR-155, miR-191, miR-218, and miR-491 were most prominently increased and that the levels of miR-200b, miR-200c, miR-26, and miR-34c were decreased. The qRT-PCR results confirm that the circulating levels of miR-21, miR-145, miR-155, and miR-191 are increased in patients with arsenism caused by coal-burning. KEGG analyses show that these miRNAs inhibit the target genes of pathways related to immune inflammation, oxidative stress, and DNA damage repair. Therefore, the four miRNAs may be biomarkers of endemic arsenism caused by coal-burning. Further studies with larger samples should be performed to confirm these findings and to elucidate the underlying mechanisms.


Somatostatin analog: effects on hypergastrinemia and hypercalcitoninemia.
A somatostatin analog (SMS 201-995) was used to treat symptomatic patients with a residual tumor burden of gastrinoma or medullary thyroid carcinoma and pathologic elevations of circulating marker peptides associated with these neuroendocrine tumors. Possible inhibitory effects of the analog on marker peptides, patients' symptoms, or tumor progression were studied in a dose-response protocol and during several months of self-injection of SMS 201-995. Both patients reported remarkable relief of secretory diarrhea and other symptoms, and serum gastrin was successfully suppressed by increasing doses of the analog. However, no effect was seen in reduction of hypercalcitoninemia. Morphologic imaging of residual tumor showed no progression of medullary thyroid carcinoma during treatment and, in the case of hepatic gastrinoma metastases, remarkable tumor regression was confirmed. No toxicity or glucose intolerance was experienced. Somatostatin analog shows promise for palliative management of endocrinologic symptoms due to neuroendocrine tumors, and an inhibitory effect can be measured in some but not all peptide markers. Further evidence of its negative trophic effect on tumor blood flow may suggest an antineoplastic potential, as well as palliative use of this new treatment.


Role of sucrose in the fitness of Streptococcus mutans.
Dental caries has been closely linked to fermentable carbohydrates as key environmental factors. Sucrose has been identified as the most cariogenic carbohydrate. Streptococcus mutans, considered to be the primary pathogen causing dental caries, is able to utilize sucrose as a nutrient source, partially for the production of intracellular storage components and for the production of extracellular glucans via the glucosyltransferases GtfB, GtfC, and GtfD. The following study explores the competitiveness and fitness of S. mutans when grown with different concentrations of sucrose. Growth competition with oral streptococci and antimicrobial susceptibility in static biofilm models grown without sucrose or with 0.1% or 0.5% sucrose were investigated using confocal laser scanning microscopy. The numbers of surviving S. mutans of both wild-type and an isogenic Gtf-negative mutant after antimicrobial treatment were determined as colony-forming units. S. mutans was able to establish microcolonies with increasing sucrose concentration in the presence of other streptococcal competitors during biofilm development. The antimicrobial susceptibility decreased when sucrose was available as substrate and was dependent on the presence of the Gtfs. The increased resistance against antimicrobial treatment was associated with the availability of sucrose, but was not influenced much by the concentration used during this study. The resistance was strongly associated with the Gtf activity, excluding any intracellular metabolic effect of sucrose in the resistance mechanism.


The Phytotherapeutic Fenugreek as Trigger of Toxic Epidermal Necrolysis.
We describe the case of a 32-year-old woman who presented to the hospital with generalized painful exanthema, blisters and erosions 1 month after giving birth to a healthy girl. The patient's medical history was inconspicuous for comorbidities; however, it included the incidental intake of pain killers and a herbal preparation (fenugreek), which she took regularly over the last 4 weeks to improve lactation. Based on the clinical characteristics, we suspected toxic epidermal necrolysis (TEN), a severe cutaneous adverse drug reaction, which was confirmed by skin biopsy. The patient was treated with high-dose intravenous human immunoglobulins and was discharged 2 weeks after hospital admission in good condition. The allergological workup identified fenugreek as the most likely causative agent. Given the increased self-medication of freely available phytotherapeutics by patients in industrialized countries, herbal mixtures should be taken into consideration in the diagnostic workup of TEN.


Challenges in Parkinson's disease: restoration of the nigrostriatal dopamine system is not enough.
Levodopa remains the most effective treatment for Parkinson's disease (PD). However, the drug is complicated by a wide range of adverse effects, most notably motor fluctuations and dyskinesias. Long-acting dopamine agonists are associated with a reduced incidence of these complications and modern surgical approaches and pharmacological methods of providing more continuous dopaminergic stimulation have a substantial ameliorative effect on these problems. Despite these advances, disease progression remains unaffected. For this reason there has been much enthusiasm for cellular therapies designed to replace degenerating nigrostriatal dopaminergic neurons. However, recent fetal transplant trials have failed to show expected benefit and have been complicated by medication dyskinesias". Even if successful, such treatment may be predestined to provide no better outcome than available treatments given current medical and surgical experience that emphasises the increasingly critical role of "non-dopaminergic" symptoms to quality of life in late-stage PD. Knowledge of the widespread, multisystem nature of the neurodegeneration that accounts for these problems suggests that restoration of the nigrostriatal dopamine system should not be the ultimate goal of future research.


Evaluation of asbestos exposure during brake repair and replacement.
Occupational exposure to asbestos fiber of brake repair job (auto mechanics) has seldom been evaluated in Iran. Accordingly, we evaluated asbestos fiber concentrations in the breathing zone of auto mechanics between July 2008 and December 2008. The asbestos fiber concentrations of 60 personal air samples collected from 30 cars and trucks brake replacement and they were analyzed by phase-contrast optical microscopy (PCM) and scanning electron microscopy (SEM) by energy-dispersive X-ray analysis. The geometric means of the personal monitoring fiber concentrations were 0.92 PCM f/ml and 0.46 PCM f/ml respectively in car and passenger heavy truck auto shops. There was a significant differences in the asbestos fiber concentrations between the car and truck auto shops (p=0.006). Based on these findings, auto mechanics who worked with asbestos containing brake may have been exposed to asbestos concentrations approximately 7 times higher than the current occupational safety and health agency (OSHA) permissible exposure limit (PEL) of 0.1 f/ml. Fiber morphology and energy dispersive X-ray analysis by SEM revealed that amphibole fibers such as tremolite and actinolite existed in the brakes dust and that the vast majority (>30%) of the airborne chrysotile fibers were greater than 1 μm in diameter. It can be concluded that the imported chrysotile asbestos contains trace amounts of tremolite and actinolite fibers and they are responsible for the high airborne asbestos levels and occupational exposure to amphibole asbestos in auto mechanics job in Iran. Thus, it is to be expected that the auto mechanics will suffer negative health effects due to exposure to the serpentine and amphibole asbestos fibers.


Sustained Release of Poorly Water-Soluble Drug from Hydrophilic Polymeric Film Sandwiched Between Hydrophobic Layers.
This proof-of-concept study explores the feasibility of using a drug-loaded hydrophilic polymeric layer sandwiched between two hydrophobic layers for improving film drug load while achieving sustained release of poorly water-soluble drug. Such films having total thickness in range ~ 146-250 μm were prepared by slurry-based casting using hydrophilic hydroxypropyl methylcellulose (HPMC) as matrix layer containing fenofibrate (FNB) as the model drug, encased between two very thin rate-limiting layers of 10 μm each of hydrophobic poly-ɛ-caprolactone (PCL). Film precursor slurry consisted of HPMC with plasticizer and water along with micronized FNB powders, which were dry-coated with hydrophilic silica. Characterization techniques demonstrated the presence of homogeneously dispersed crystalline FNB in films. The films are very thin and hence two-dimensional; hence, average drug load per unit area in range ~ 5 to ~ 9 mg/cm2 could be achieved by altering the thickness of the drug matrix layer. Drug amount and drug content uniformity were measured through assay of ten circular samples ~ 0.712 cm2 in area punched out using a circular-shaped punch tool. Drug release rate was investigated using USP IV flow-through cell and surface dissolution imaging system. Thinner films followed Fickian diffusion, and thicker films followed non-Fickian anomalous diffusion. Overall, the application of middle layer thickness could be used as a tool to manipulate drug load without the need for altering its formulation or precursor preparation by changing its thickness, hence achieving relatively high drug loading yet having sustained release of drug.


Thysanoptera (thrips) within citrus orchards in Florida: species distribution, relative and seasonal abundance within trees, and species on vines and ground cover plants.
Seven citrus orchards on reduced to no pesticide spray programs were sampled for Thysanoptera in central and south central Florida. Inner and outer canopy leaves, fruits, twigs, trunk scrapings, vines and ground cover plants were sampled monthly between January 1995 and January 1996. Thirty-six species of thrips were identified from 2,979 specimens collected from within citrus tree canopies and 18,266 specimens from vines and ground cover plants within the seven citrus orchards. The thrips species included seven predators [Aleurodothrips fasciapennis (Franklin), Karnyothrips flavipes (Jones), K. melaleucus (Bagnall), Leptothrips cassiae (Watson), L. macroocellatus (Watson), L. pini (Watson), and Scolothrips sexmaculatus (Pergande)] 21 plant feeding species [Anaphothrips n. sp., Arorathrips mexicanus (Crawford), Aurantothrips orchidaceous (Bagnall), Baileyothrips limbatus (Hood), Chaetanaphothrips orchidii (Moulton), Danothrips trifasciatus (Sakimura), Echinothrips americanus (Morgan), Frankliniella bispinosa (Morgan), F. cephalica (Crawford), F. fusca (Hinds), F. gossypiana (Hood), Frankliniella sp. (runneri group), Haplothrips gowdeyi (Franklin), Heliothrips haemorrhoidalis (Bouché), Leucothrips piercei (Morgan), Microcephalothrips abdominalis (Crawford), Neohydatothrips floridanus (Watson), N. portoricensis (Morgan), Pseudothrips inequalis (Beach), Scirtothrips sp., and Thrips hawaiiensis (Morgan)]; and eight fungivorous feeding species [Adraneothrips decorus (Hood), Hoplandrothrips pergandei (Hinds), Idolothripinae sp., Merothrips floridensis (Watson), M. morgani (Hood), Neurothrips magnafemoralis (Hinds), Stephanothrips occidentalis Hood and Williams, and Symphyothrips sp.]. Only F. bispinosa, C. orchidii, D. trifasciatus, and H. haemorrhoidalis have been considered economic pests on Florida citrus. Scirtothrips sp. and T. hawaiiensis were recovered in low numbers within Florida citrus orchards. Both are potential pest species to citrus and possibly other crops in Florida. The five most abundant thrips species collected within citrus tree canopies were: A. fasciapennis, F. bispinosa, C. orchidii, K. flavipes, and D. trifasciatus. In comparison, the following five thrips species were most abundant on vines or ground cover plants: F. bispinosa, H. gowdeyi, F. cephalica, M. abdominalis, and F. gossypiana. Fifty-eight species of vines or ground cover plants in 26 families were infested with one or more of 27 species of thrips.


Human subthalamic neuron spiking exhibits subtle responses to sedatives.
During deep brain stimulation implant surgery, microelectrode recordings are used to map the location of targeted neurons. The effects produced by propofol or remifentanil on discharge activity of subthalamic neurons were studied intraoperatively to determine whether they alter neuronal activity. Microelectrode recordings from 11 neurons, each from individual patients, were discriminated and analyzed before and after administration of either propofol or remifentanil. Subthalamic neurons in rat brain slices were recorded in patch-clamp to investigate cellular level effects. Neurons discharged at 42 ± 9 spikes/s (mean ± SD) and showed a common pattern of inhibition that lasted 4.3 ms. Unique discharge profiles were evident for each neuron, seen using joint-interval analysis. Propofol (intravenous bolus 0.3 mg/kg) produced sedation, with minor effects on discharge activity (less than 2.0% change in frequency). A prolongation of recurrent inhibition was evident from joint-interval analysis, and propofol's effect peaked within 2 min, with recovery evident at 10 min. Subthalamic neurons recorded in rat brain slices exhibited inhibitory synaptic currents that were prolonged by propofol (155%) but appeared to lack tonic inhibitory currents. Propofol did not alter membrane potential, membrane resistance, current-evoked discharge, or holding current during voltage clamp. Remifentanil (0.05 mg/kg) had little effect on overall subthalamic neuron discharge activity and did not prolong recurrent inhibition. These results help to characterize the circuit properties and feedback inhibition of subthalamic neurons and demonstrate that both propofol and remifentanil produce only minor alterations of subthalamic neuron discharge activity that should not interfere with deep brain stimulation implant surgery.


Lipophilicity and Click Reactivity Determine the Performance of Bioorthogonal Tetrazine Tools in Pretargeted In Vivo Chemistry.
The development of highly selective and fast biocompatible reactions for ligation and cleavage has paved the way for new diagnostic and therapeutic applications of pretargeted in vivo chemistry. The concept of bioorthogonal pretargeting has attracted considerable interest, in particular for the targeted delivery of radionuclides and drugs. In nuclear medicine, pretargeting can provide increased target-to-background ratios at early time-points compared to traditional approaches. This reduces the radiation burden to healthy tissue and, depending on the selected radionuclide, enables better imaging contrast or higher therapeutic efficiency. Moreover, bioorthogonally triggered cleavage of pretargeted antibody-drug conjugates represents an emerging strategy to achieve controlled release and locally increased drug concentrations. The toolbox of bioorthogonal reactions has significantly expanded in the past decade, with the tetrazine ligation being the fastest and one of the most versatile in vivo chemistries. Progress in the field, however, relies heavily on the development and evaluation of (radio)labeled compounds, preventing the use of compound libraries for systematic studies. The rational design of tetrazine probes and triggers has thus been impeded by the limited understanding of the impact of structural parameters on the in vivo ligation performance. In this work, we describe the development of a pretargeted blocking assay that allows for the investigation of the in vivo fate of a structurally diverse library of 45 unlabeled tetrazines and their capability to reach and react with pretargeted trans-cyclooctene (TCO)-modified antibodies in tumor-bearing mice. This study enabled us to assess the correlation of click reactivity and lipophilicity of tetrazines with their in vivo performance. In particular, high rate constants (>50 000 M-1 s-1) for the reaction with TCO and low calculated logD 7.4 values (below -3) of the tetrazine were identified as strong indicators for successful pretargeting. Radiolabeling gave access to a set of selected 18F-labeled tetrazines, including highly reactive scaffolds, which were used in pretargeted PET imaging studies to confirm the results from the blocking study. These insights thus enable the rational design of tetrazine probes for in vivo application and will thereby assist the clinical translation of bioorthogonal pretargeting.


Associations between motor proficiency and academic performance in mathematics and reading in year 1 school children: a cross-sectional study.
A key priority for learning during the early years of school is for children to develop skills in numeracy and literacy. Consequently, less time may be allocated in the curriculum to foster other important developmental areas, including the ongoing motor skill development of school children, which has been positively linked to academic performance. In order to promote holistic approaches to teaching and learning in the early years of school, it is necessary to further delineate the nature of associations between motor skills and foundation academic skills. The aim of this study was to examine associations between fine and gross motor proficiency and academic performance in mathematics and reading in Year 1 children. A cross-sectional study was conducted with Year 1 children from two primary schools in New South Wales, Australia (N = 55; 25 boys, 30 girls; mean age 6.77 ± 0.40 years). The Bruininks-Oseretsky Test of Motor Proficiency (2nd Edition) and the Wechsler Individual Achievement Test II (Australian Edition) were used to assess motor proficiency and academic performance in mathematics and reading, respectively. Associations between the components of motor proficiency and academic outcomes were examined using Pearson's and Spearman's correlation analyses. Hierarchical multiple linear regression analyses were conducted to determine how much variance in mathematics and reading composite scores could be explained by motor proficiency after controlling for age. A significant moderate positive association was found between total motor composite and mathematics composite scores (r = .466, p < .001). Fine manual control composite scores were significantly associated with both mathematics (rs = .572, p < .001) and reading (rs = .476, p = .001) composite scores. After controlling for age, fine motor integration was the only component of motor proficiency that explained significant variance in mathematics and reading composite scores. The results of the study revealed that Year 1 children's overall motor proficiency was significantly related to their mathematical ability. Children's fine motor integration skills were also predictive of mathematics and reading ability. These study findings may interest both early childhood educators and paediatric health professionals.


Synthesis and biological evaluation of chalcone-linked pyrazolo[1,5-a]pyrimidines as potential anticancer agents.
A series of pyrazolo[1,5-a]pyrimidines substituted at C5 with 1-phenylprop-2-en-1-one (6a-q) and 3-phenylprop-2-en-1-one (7a-k) was synthesized and evaluated for antiproliferative activity. Among them, 6h was found to be the most active compound against the MDA-MB-231 cell line with an IC50 of 2.6 μM . The antiproliferative activity of this series of compounds ranged from 2.6 to 34.9 μM against A549 (lung cancer), MDA-MB-231 (breast cancer) and DU-145 (prostate cancer) cell lines. FACS analysis revealed that these hybrids arrest the cell cycle at the subG1 phase. Western blot analysis and an immunofluorescence assay showed the inhibition of the EGFR and STAT3 axis, which plays an important role in cell survival and apoptosis. Western blot and RT-PCR analyses that displayed an increase in apoptotic proteins such as p53, p21 and Bax and a decrease in the anti-apoptotic proteins Bcl-2 and procaspase-9 confirmed the ability of these hybrids to trigger cell death by apoptosis. Molecular docking studies described the binding of these hybrids to the ATP binding site of EGFR.


The role of Zn2+ in Shal voltage-gated potassium channel formation.
Voltage-gated potassium channels are formed by the tetramerization of their alpha subunits, in a process that is controlled by their conserved N-terminal T1 domains. The crystal structures of Shaker and Shaw T1 domains reveal interesting differences in structures that are contained within a highly conserved BTB/POZ domain fold. The most surprising difference is that the Shaw T1 domain contains an intersubunit Zn2+ ion that is lacking in the Shaker T1 domain. The Zn2+ coordination motif is conserved in other non-Shaker channels making this the most distinctive difference between these channels and Shaker. In this study we show that Zn2+ is an important co-factor for the tetramerization of isolated Shaw and Shal T1 domains. Addition of Zn2+ increases the amount of tetramer formed, whereas chelation of Zn2+ with phenanthroline blocks tetramerization and causes assembled tetramers to disassemble. Within an intact cell, full-length Shal subunits containing Zn2+ site mutations also fail to form functional channels, with the majority of the protein found to remain monomeric by size exclusion chromatography. Therefore, zinc-mediated tetramerization also is a physiologically important event for full-length functional channel formation.


Low-density lipoprotein receptor-related protein 1 (LRP1)-dependent cell signaling promotes axonal regeneration.
Low-density lipoprotein receptors (LRPs) are present extensively on cells outside of the nervous system and classically exert roles in lipoprotein metabolism. It has been reported recently that LRP1 activation could phosphorylate the neurotrophin receptor TrkA in PC12 cells and increase neurite outgrowth from developing cerebellar granule cells. These intriguing findings led us to explore the hypothesis that LRP1 activation would activate canonical neurotrophic factor signaling in adult neurons and promote axonal regeneration after spinal cord injury. We now find that treatment of adult rat dorsal root ganglion neurons in vitro with LRP1 agonists (the receptor binding domain of α-2-macroglobulin or the hemopexin domain of matrix metalloproteinase 9) induces TrkC, Akt, and ERK activation; significantly increases neurite outgrowth (p < 0.01); and overcomes myelin inhibition (p < 0.05). These effects require Src family kinase activation, a classic LRP1-mediated Trk transactivator. Moreover, intrathecal infusions of LRP1 agonists significantly enhance sensory axonal sprouting and regeneration after spinal cord injury in rats compared with control-infused animals (p < 0.05). A significant role is established for lipoprotein receptors in sprouting and regeneration after CNS injury, identifying a novel class of therapeutic targets to explore for traumatic neurological disorders.


Tissue distribution and in vivo immunosuppressive activity of liposomal cyclosporine.
The distribution and immunosuppressive activity of liposomal cyclosporine (L-CSA) when administered iv as single- and multiple-doses were compared with the commercially available cyclosporine (C-CSA) in rats. CSA concentrations in the spleen and liver were significantly greater 1 hr after dosing in rats given L-CSA compared with C-CSA. The apparent tissue to blood partition coefficient at 1 hr after dosing was significantly greater for the liver and spleen of rats treated with L-CSA compared with C-CSA. Drug concentrations 24 hr after single doses of L-CSA were significantly lower in the kidney, heart, lung, and adipose tissues compared with animals given C-CSA. Lymphocyte-blastogenic response was studied in a separate group of rats given 10 mg/kg/day of L-CSA or C-CSA for 10 days compared with drug-free control groups. A 3-fold decrease in blastogenic response was observed in rats given L-CSA compared with C-CSA-treated rats (12.7 +/- 11.8 vs. 34.9 +/- 11.3 x 10(3) dpm/10(6) cells; p less than 0.05). These data suggest that the liposomal formulation of CSA leads to enhanced tissue levels of CSA in the spleen coupled with augmentation in immunosuppressive activity.


Study protocol for "Study of Practices Enabling Implementation and Adaptation in the Safety Net (SPREAD-NET)": a pragmatic trial comparing implementation strategies.
Little research has directly compared the effectiveness of implementation strategies in any setting, and we know of no prior trials directly comparing how effectively different combinations of strategies support implementation in community health centers. This paper outlines the protocol of the Study of Practices Enabling Implementation and Adaptation in the Safety Net (SPREAD-NET), a trial designed to compare the effectiveness of several common strategies for supporting implementation of an intervention and explore contextual factors that impact the strategies' effectiveness in the community health center setting. This cluster-randomized trial compares how three increasingly hands-on implementation strategies support adoption of an evidence-based diabetes quality improvement intervention in 29 community health centers, managed by 12 healthcare organizations. The strategies are as follows: (arm 1) a toolkit, presented in paper and electronic form, which includes a training webinar; (arm 2) toolkit plus in-person training with a focus on practice change and change management strategies; and (arm 3) toolkit, in-person training, plus practice facilitation with on-site visits. We use a mixed methods approach to data collection and analysis: (i) baseline surveys on study clinic characteristics, to explore how these characteristics impact the clinics' ability to implement the tools and the effectiveness of each implementation strategy; (ii) quantitative data on change in rates of guideline-concordant prescribing; and (iii) qualitative data on the "how" and "why" underlying the quantitative results. The outcomes of interest are clinic-level results, categorized using the Reach, Effectiveness, Adoption, Implementation, Maintenance (RE-AIM) framework, within an interrupted time-series design with segmented regression models. This pragmatic trial will compare how well each implementation strategy works in "real-world" practices. Having a better understanding of how different strategies support implementation efforts could positively impact the field of implementation science, by comparing practical, generalizable methods for implementing clinical innovations in community health centers. Bridging this gap in the literature is a critical step towards the national long-term goal of effectively disseminating and implementing effective interventions into community health centers. ClinicalTrials.gov, NCT02325531.


3'-5' exonuclease activity of human apurinic/apyrimidinic endonuclease 1 towards DNAs containing dNMP and their modified analogs at the 3 end of single strand DNA break.
Human DNA apurinic/apyrimidinic (AP-) endonuclease 1 (APE1) is involved in the base excision repair (BER) pathway. The enzyme hydrolyzes DNA from the 5 side of the AP site. In addition to endonuclease activity, APE1 also possesses other slight activities including 3 -5 exonuclease activity. The latter is preferentially exhibited towards mispaired (non-canonical) nucleotides, this being the reason why APE1 is considered as a proofreading enzyme correcting the misincorporations introduced by DNA polymerase beta. We have studied 3 -5 exonuclease activity of APE1 towards dCMP and dTMP residues and modified dCMP analogs with photoreactive groups at the 3 end of the nicked DNA. Photoreactive dNMP residues were incorporated at the 3 end of the lesion using DNA polymerase beta and photoreactive dNTPs. The dependence of exonuclease activity on the "canonicity" of the base pair formed by dNMP flanking the nick at the 3 end, on the nature of the group flanking the nick at the 5 end, and on the reaction conditions has been determined. Optimal reaction conditions for the 3 -5 exonuclease hydrolysis reaction catalyzed by APE1 in vitro have been established, and conditions when photoreactive residues are not removed by APE1 have been chosen. These reaction conditions are suitable for using photoreactive nicked DNAs bearing 3 -photoreactive dNMP residues for photoaffinity labeling of proteins in cellular/nuclear extracts and model APE1-containing systems. We recommend using FAPdCTP for photoaffinity modification in APE1-containing systems because the FAPdCMP residue is less prone to exonuclease degradation, in contrast to FABOdCTP, which is not recommended.


Identification and in silico analyses of novel TGFBR1 and TGFBR2 mutations in Marfan syndrome-related disorders.
Very recently, heterozygous mutations in the genes encoding transforming growth factor beta receptors I (TGFBR1) and II (TGFBR2) have been reported in Loeys-Dietz aortic aneurysm syndrome (LDS). In addition, dominant TGFBR2 mutations have been identified in Marfan syndrome type 2 (MFS2) and familial thoracic aortic aneurysms and dissections (TAAD). In the past, mutations of these genes were associated with atherosclerosis and several human cancers. Here, we report a total of nine novel and one known heterozygous sequence variants in the TGFBR1 and TGFBR2 genes in nine of 70 unrelated individuals with MFS-like phenotypes who previously tested negative for mutations in the gene encoding the extracellular matrix protein fibrillin-1 (FBN1). To assess the pathogenic impact of these sequence variants, in silico analyses were performed by the PolyPhen, SIFT, and Fold-X algorithms and by means of a 3D homology model of the TGFBR2 kinase domain. Our results showed that in all but one of the patients the pathogenic effect of at least one sequence variant is highly probable (c.722C > T, c.799A > C, and c.1460G > A in TGFBR1 and c.773T > G, c.1106G > T, c.1159G > A, c.1181G > A, and c.1561T > C in TGFBR2). These deleterious alleles occurred de novo or segregated with the disease in the families, indicating a causative association between the sequence variants and clinical phenotypes. Since TGFBR2 mutations found in patients with MFS-related disorders cannot be distinguished from heterozygous TGFBR2 mutations reported in tumor samples, we emphasize the importance of segregation analysis in affected families. In order to be able to find the mutation that is indeed responsible for a MFS-related phenotype, we also propose that genetic testing for sequence alterations in TGFBR1 and TGFBR2 should be complemented by mutation screening of the FBN1 gene.


Therapeutic Potential of Hericium erinaceus for Depressive Disorder.
Depression is a common and severe neuropsychiatric disorder that is one of the leading causes of global disease burden. Although various anti-depressants are currently available, their efficacies are barely adequate and many have side effects. Hericium erinaceus, also known as Lion's mane mushroom, has been shown to have various health benefits, including antioxidative, antidiabetic, anticancer, anti-inflammatory, antimicrobial, antihyperglycemic, and hypolipidemic effects. It has been used to treat cognitive impairment, Parkinson's disease, and Alzheimer's disease. Bioactive compounds extracted from the mycelia and fruiting bodies of H. erinaceus have been found to promote the expression of neurotrophic factors that are associated with cell proliferation such as nerve growth factors. Although antidepressant effects of H. erinaceus have not been validated and compared to the conventional antidepressants, based on the neurotrophic and neurogenic pathophysiology of depression, H. erinaceus may be a potential alternative medicine for the treatment of depression. This article critically reviews the current literature on the potential benefits of H. erinaceus as a treatment for depressive disorder as well as its mechanisms underlying the antidepressant-like activities.


HPV and HPV vaccination: knowledge and consciousness of young women.
The aim of the study was to explore the knowledge and the awareness of the young Turkish women regarding cervical cancer and human papilloma virus (HPV) vaccines. The authors analyze a probable relationship between the overall knowledge level and a few socio-demographic parameters. The authors interviewed with students from Canakkale 18 March University and young women that did not continue with school in the same city from January to September 2011. All the students answered the questionnaire voluntarily and independently. The participants had low level of knowledge about the risk factors for cervical cancer. Smoking is the major risk factor that was known by the participants (65%). Proportion of the participants that were aware of pap smear test and HPV were 65% and 17% respectively. A small proportion of young women had knowledge regarding protection from HPV. Educational stream, educational level, family income, and family size had significant association knowledge level (p < 0.05). There has not been any improvement in HPV and risk factor of cervical cancer awareness in young women. Health members of the National Cancer Control Programme and delegates of the vaccine corporations have major work in order to increase the level of knowledge so that general public can easily take preventative measures.


Randomized blinded controlled trial of dipyrone as a treatment for pyrexia in horses.
OBJECTIVE To evaluate the effectiveness and safety of dipyrone to control pyrexia in horses with naturally occurring disease under field conditions. ANIMALS 138 horses with pyrexia and various infections evaluated at 14 veterinary sites in 12 states. PROCEDURES In the first (effectiveness) phase of this 2-phase study, horses were randomly assigned 3:1 to receive 1 dose of dipyrone (30 mg/kg [13.6 mg/lb], IV) or an equivalent amount of placebo. Effectiveness was defined as a decrease in rectal temperature ≥ 1.1°C (2°F), compared with the pretreatment value, or a rectal temperature of ≤ 38.3°C (101.0°F) 6 hours after treatment administration. Horses deemed to have an appropriate reduction in rectal temperature (regardless of treatment group) by 6 hours were immediately entered into the safety phase of the study, in which dipyrone was administered IV at 30 mg/kg between 0 and 8 times up to every 8 hours on an as-needed basis, as determined by the clinical investigators. Horses were monitored throughout for adverse events. RESULTS A significantly greater proportion of dipyrone-treated horses (76/99 [77%]) had an effective treatment response than did placebo-treated horses (6/31 [19%]). Posttreatment adverse events were mild and transient. No differences in types or prevalence of gastrointestinal adverse events were evident between treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE Dipyrone was effective in controlling pyrexia by 6 hours after IV administration of a single 30-mg/kg dose in a large proportion of treated horses. Adverse effects were minimal.


First Report of Leaf Spot Caused by Xanthomonas axonopodis pv. poinsettiicola on Poinsettia (Euphorbia pulcherrima) in Norway.
Poinsettia (Euphorbia pulcherrima) is the biggest flowering potted-plant culture in Norway with approximately six million plants produced yearly. A considerable percentage is produced from imported cuttings. In September 2010, we received diseased poinsettias with necrotic leaf spots from a commercial poinsettia grower in Hordaland County. Leaf spots on the upper sides of leaves were brownish, necrotic, irregular in shape, and surrounded by yellow halos. Small, grayish brown, water-soaked spots were observed on the abaxial sides of the leaves from the same plants. Some of the latter carried crusty, dried residues of bacterial exudates. Leaves were surface sterilized and small pieces were excised from the transition area between healthy and diseased tissue. Leaf pieces were soaked in 0.2 ml of sterile phosphate buffered saline (SPBS) for 30 min. The resulting solution was diluted and streaked on several common media suited for the recovery of plant pathogenic bacteria, among them YDC (yeast dextrose chalk agar). The plates were incubated at 25°C in the dark. After 48 to 72 h, pale yellow, smooth, convex, round, and shiny colonies appeared on YDC. On the basis of plant symptoms and colony morphology, the isolated bacteria were expected to be Xanthomonas axonopodis pv poinsettiicola, which is a known pathogen of poinsettia. One isolate was analyzed by fatty acid methyl ester (FAME) analysis according to Sasser (2) and partial gyrase B sequencing as described by Ah-You et al. (1). A strain of X. axonopodis pv. poinsettiicola (NCPPB 581) from the National Collection of Plant Pathogenic Bacteria (UK) was included as a control in both analyses. The isolates were identical to NCPPB581 with respect to the FAME analysis (species level) and the gyrase B sequence. Furthermore, the gyrB sequence was identical to the sequence of strain LMG 849 in GenBank (Accession No. EU015342.1; identities = 774 of 774). Leaf inoculation of disease-free poinsettia was carried out by spraying a solution (approximately 108 CFU ml-1) on the leaves, covering the plants with wetted plastic bags, and placing the plants in a greenhouse maintained at 21°C for 4 weeks. Leaf spot symptoms consistent with the previously observed ones appeared after 2 weeks of incubation. No symptoms were observed on the negative control plant, which was sprayed with SPBS only. The bacterium was successfully reisolated from the induced symptoms and identified by FAME analysis and gyrase B sequencing. In the period following the first detection, Norwegian poinsettia growers were advised to inspect their produce. Suspected samples were sent to us from 28 producers from around the country. The pathogen was detected at 15 production places. Growers were recommended to disinfect their premises and be vigilant with respect to starting up the new season with healthy propagation material. To our knowledge, this is the first report of X. axonopodis pv. poinsettiicola causing bacterial leaf spot on poinsettia in Norway, providing further data on the occurrence of the disease in Europe. References: (1) N. Ah-You et al. Int. J. Syst. Evol. Microbiol. 59:306, 2009. (2) M. J. Sasser. MIDI Tech. Note No. 102. MIDI, 115 Barksdale Prof. Center, Newark, DE, 1990.


FMRFamide-related peptides in potato cyst nematodes.
This study presents data demonstrating the presence of FMRFamide-related peptides (FaRPs) in potato cyst nematodes (PCN). Five transcripts of FaRP encoding genes, designated gpflp-1 to gpflp-5, were characterised using RACE. In terms of ORFs, gpflp-1 was 444 base pairs (bp) long and coded for four copies of the FaRP, PF3 (KSAYMRFamide) whilst gpflp-2 was 309 bp long and encoded one copy of the peptide, KNKFEFIRFamide. gpflp-3 (420 bp) Encoded two copies of KHEYLRFamide (AF2) and the genes gpflp-4 and gpflp-5 encoded a total of 11 FaRPs, most of which are novel to PCN. FMRFamide-related peptide (FaRP)-like immunoreactivity was observed in both PCN species, Globodera pallida and Globodera rostochiensis, using an antiserum raised against the invertebrate peptide, FMRFamide. Immunopositive neurones were found throughout the central nervous system in the ventral and dorsal nerve cords and the circumpharyngeal and perianal nerve rings. Reactive neurones were also present peripherally, innervating the highly muscular pharynx with a nerve net and ring-like structures. Positive immunostaining was also observed in neurones running toward the stylet protractor muscles and/or the anterior sensory apparatus. This study implicates a role for FaRPs in feeding, host penetration and sensory function of PCN. This is the first study to characterise FaRP encoding genes from a plant-parasitic nematode using a targeted PCR based RACE approach and further underlines the importance and diversity of this neuropeptide group in the phylum Nematoda.


Host cell membrane microdomains and fungal infection.
Lipid microdomains or lipid rafts are dynamic and tightly ordered regions of the plasma membrane. In mammalian cells, they are enriched in cholesterol, glycosphingolipids, Glycosylphosphatidylinositol-anchored and signalling-related proteins. Several studies have suggested that mammalian pattern recognition receptors are concentrated or recruited to lipid domains during host-pathogen association to enhance the effectiveness of host effector processes. However, pathogens have also evolved strategies to exploit these domains to invade cells and survive. In fungal organisms, a complex cell wall network usually mediates the first contact with the host cells. This cell wall may contain virulence factors that interfere with the host membrane microdomains dynamics, potentially impacting the infection outcome. Indeed, the microdomain disruption can dampen fungus-host cell adhesion, phagocytosis and cellular immune responses. Here, we provide an overview of regulatory strategies employed by pathogenic fungi to engage with and potentially subvert the lipid microdomains of host cells. TAKE AWAY: Lipid microdomains are ordered regions of the plasma membrane enriched in cholesterol, glycosphingolipids (GSL), GPI-anchored and signalling-related proteins. Pathogen recognition by host immune cells can involve lipid microdomain participation. During this process, these domains can coalesce in larger complexes recruiting receptors and signalling proteins, significantly increasing their signalling abilities. The antifungal innate immune response is mediated by the engagement of pathogen-associated molecular patterns to pattern recognition receptors (PRRs) at the plasma membrane of innate immune cells. Lipid microdomains can concentrate or recruit PRRs during host cell-fungi association through a multi-interactive mechanism. This association can enhance the effectiveness of host effector processes. However, virulence factors at the fungal cell surface and extracellular vesicles can re-assembly these domains, compromising the downstream signalling and favouring the disease development. Lipid microdomains are therefore very attractive targets for novel drugs to combat fungal infections.


Regulation of bone formation by baicalein via the mTORC1 pathway.
Osteoporosis is a systemic skeletal disease that is characterized by low bone density and microarchitectural deterioration of bone tissue. The increasing prevalence of osteoporosis has attracted much attention. In this study, MC3T3-E1 pre-osteoblasts were treated with the natural compound, baicalein (0.1 μmol/L, 1 μmol/L, 10 μmol/L), to stimulate differentiation over a 14-day period. In addition, a canonical ovariectomized (OVX) mouse model was used to investigate the effect of 3-month baicalein treatment (10 mg/kg per day) in preventing postmenopausal osteoporosis. In vitro, we found that baicalein induced activation of alkaline phosphatase, stimulated the mammalian target of rapamycin complex 1 (mTORC1) signaling pathway, and induced expression of osteoblast differentiation markers, ie, osteocalcin, osterix, collagen Iα1, and runt-related transcription factor 2 (RUNX2), in osteoblasts. In vivo, several bone parameters, including trabecular thickness, trabecular bone mineral density, and trabecular number, in the distal femoral metaphysis were significantly increased in OVX mice treated intragastrically with baicalein for 3 months compared with OVX mice that were not treated with baicalein. We also found that expression of osteocalcin and RUNX2 was decreased in primary ossified tissue from the OVX group, and baicalein increased the levels of osteocalcin and RUNX2 in OVX mice. These data suggest that baicalein can stimulate MC3T3-E1 cells to differentiate into osteoblasts via activation of the mTORC1 signaling pathway, which includes protein kinases and transcription factors such as P-4E/BP1 and P-S6K1.


Physiological Basis and Transcriptional Profiling of Three Salt-Tolerant Mutant Lines of Rice.
Salinity is a complex trait that affects growth and productivity in many crops, including rice. Mutation induction, a useful tool to generate salt tolerant plants, enables the analysis of plants with similar genetic background, facilitating the understanding of the salt tolerance mechanisms. In this work, we generated three salt tolerant mutant lines by irradiation of a salt-sensitive cultivar plants and screened M2 plants at seedling stage in the presence of high salinity. These three lines, SaT20, SaS62, and SaT58, showed different responses to salinity, but exhibited similar phenotype to wild type plants, except SaT20 that displayed shorter height when grown in the absence of salt. Under salt conditions, all three mutants and the parental line showed similar reduction in yield, although relevant differences in other physiological parameters, such as Na+ accumulation in healthy leaves of SaT20, were registered. Microarray analyses of gene expression profiles in roots revealed the occurrence of common and specific responses in the mutants. The three mutants showed up-regulation of responsive genes, the activation of oxido-reduction process and the inhibition of ion transport. The participation of jasmonate in the plant response to salt was evident by down-regulation of a gene coding for a jasmonate O-methyltransferase. Genes dealing with lipid transport and metabolism were, in general, up-regulated except in SaS62, that also exhibited down-regulation of genes involved in ion transport and Ca2+ signal transduction. The two most tolerant varieties, SaS62 and SaT20, displayed lower levels of transcripts involved in K+ uptake. The physiological study and the description of the expression analysis evidenced that the three lines showed different responses to salt: SaT20 showed a high Na+ content in leaves, SaS62 presented an inhibition of lipid metabolism and ion transport and SaT58 differs in both features in the response to salinity. The analysis of these salt tolerant mutants illustrates the complexity of this trait evidencing the breadth of the plant responses to salinity including simultaneous cooperation of alternative or complementary mechanisms.


FGFR3-TACC3 fusion proteins act as naturally occurring drivers of tumor resistance by functionally substituting for EGFR/ERK signaling.
The epidermal growth factor receptor (EGFR) is a clinically validated target in head and neck squamous cell carcinoma (HNSCC), where EGFR-blocking antibodies are approved for first-line treatment. However, as with other targeted therapies, intrinsic/acquired resistance mechanisms limit efficacy. In the FaDu HNSCC xenograft model, we show that combined blockade of EGFR and ERBB3 promotes rapid tumor regression, followed by the eventual outgrowth of resistant cells. RNA sequencing revealed that resistant cells express FGFR3-TACC3 fusion proteins, which were validated as drivers of the resistant phenotype by several approaches, including CRISPR-mediated inactivation of FGFR3-TACC3 fusion genes. Interestingly, analysis of signaling in resistant cell lines demonstrated that FGFR3-TACC3 fusion proteins promote resistance by preferentially substituting for EGFR/RAS/ERK signaling rather than ERBB3/PI3K/AKT signaling. Furthermore, although FGFR3-TACC3 fusion proteins promote resistance of additional EGFR-dependent HNSCC and lung cancer cell lines to EGFR blockade, they are unable to compensate for inhibition of PI3K signaling in PIK3CA-mutant HNSCC cell lines. Validation of FGFR3-TACC3 fusion proteins as endogenous drivers of resistance in our screen provides strong evidence that these fusions are capable of substituting for EGFR signaling. Thus, FGFR3-TACC3 fusion proteins may represent a novel mechanism of acquired resistance in EGFR-dependent cancers of multiple cell lineages.


The reliability and validity of radiological assessment for patellar instability. A systematic review and meta-analysis.
To determine the discriminative validity and reliability of the evidence base using meta-analysis. A review of published sources using the databases AMED, CINHAL, EMBASE, MEDLINE, Scopus and the Cochrane Library, and for unpublished material was conducted. All studies assessing the reliability, validity, sensitivity or specificity of magnetic resonance imaging (MRI), computed tomography (CT) or ultrasound (US) of the patellofemoral joint of patients following patellar dislocation, subluxation or instability, were included. A meta-analysis was performed to assess the difference in radiological measurements between healthy controls and subjects with patellar instability in order to assess discrimination validity. A narrative assessment was used to evaluate the inter- and intra-observer reliability as well as the sensitivity and specificity of specific radiological measurements. A total of 27 studies were reviewed. The findings indicated that there was acceptable inter-observer and intra-observer reliability and validity for different methods of assessing patellar height and the sulcus angle with X-ray, MRI and CT methods, and the tibial tubercle-trochlear groove (TT-TG) assessed using CT. There was poor reliability or validity for the assessment of severity of trochlear dysplasia and the sulcus angle using US. There is insufficient evidence to determine the reliability, validity, sensitivity or specificity of tests such as the congruence angle, lateral patellar displacement, lateral patellar tilt, trochlear depth, boss height, the crossing sign or Wiberg patellar classification. A critical appraisal of the literature identified a number of recurrent methodological limitations. Further study is recommended to evaluate the reliability and validity of these radiological outcomes using well-designed radiological trials.


Qualitative Behavioural Assessment of emotionality in pigs.
Scientific assessment of affective states in animals is challenging but vital for animal welfare studies. One possible approach is Qualitative Behavioural Assessment (QBA), a 'whole animal' methodology which integrates information from multiple behavioural signals and styles of behavioural expression (body language) directly in terms of an animal's emotional expression. If QBA provides a valid measure of animals' emotional state it should distinguish between groups where emotional states have been manipulated. To test this hypothesis, QBA was applied to video-recordings of pigs, following treatment with either saline or the neuroleptic drug Azaperone, in either an open field or elevated plus-maze test. QBA analysis of these recordings was provided by 12 observers, blind to treatment, using a Free Choice Profiling (FCP) methodology. Generalised Procrustes Analysis was used to calculate a consensus profile, consisting of the main dimensions of expression. Dimension one was positively associated with terms such as 'Confident' and 'Curious' and negatively with 'Unsure' and 'Nervous'. Dimension two ranged from 'Agitated'/'Angry' to 'Calm'/'Relaxed'. In both tests, Azaperone pre-treatment was associated with a more positive emotionality (higher scores on dimension one reflecting a more confident/curious behavioural demeanour) than control pigs. No effect of drug treatment on dimension two was found. Relationships between qualitative descriptions of behaviour and quantitative behavioural measures, taken from the same recordings, were found. Overall, this work supports the use of QBA for the assessment of emotionality in animals.


Apolipoprotein E genotype and subcortical vascular lesions in older depressed patients and control subjects.
Several studies have linked geriatric depression with cerebrovascular disease. The apolipoprotein E gene (APOE) epsilon 4 allele has been associated with a variety of late-life neuropsychiatric disorders, including Alzheimer's disease, vascular dementia, and depression. The sample consisted of 145 elderly depressive individuals and 100 nondepressed elderly control subjects. After a standardized clinical assessment, all subjects underwent a magnetic resonance imaging brain scan. Volumes of subcortical white and gray matter lesions were determined using a semi-automated method. Apolipoprotein E genotype was determined on blood sample using a standard protocol. A series of linear regression models were developed to assess the relationships between APOE genotype and white and gray matter lesion volumes. Older age, lower Mini-Mental State Examination score, and having any APOE epsilon 4 allele were each correlated with gray-matter lesion volume in depressed patients. Apolipoprotein E genotype was not associated with any lesion volume among control subjects. In a subsequent linear regression model, gray matter lesion volume was associated with older age, having at least one APOE epsilon 4 allele, and white matter lesion volume among depressed patients. These results are consistent with previous reports linking cerebrovascular disease and APOE genotype. Further studies are needed to replicate this finding in elderly depressive individuals and to explain the relationship between the APOE locus and development of central nervous system vascular pathology.


Circulating tumor cells and [18F]fluorodeoxyglucose positron emission tomography/computed tomography for outcome prediction in metastatic breast cancer.
Circulating tumor cells (CTCs) and [(18)F]fluorodeoxyglucose (FDG) positron emission tomography (PET)/computed tomography (CT) are two new promising tools for therapeutic monitoring. In this study, we compared the prognostic value of CTC and FDG-PET/CT monitoring during systemic therapy for metastatic breast cancer (MBC). A retrospective analyses of 115 MBC patients who started a new line of therapy and who had CTC counts and FDG-PET/CT scans performed at baseline and at 9 to 12 weeks during therapy (midtherapy) was performed. Patients were categorized according to midtherapy CTC counts as favorable (ie, < five CTCs/7.5 mL blood) or unfavorable (> or = five CTCs/7.5 mL blood) outcomes. CTC counts and FDG-PET/CT response at midtherapy were compared, and univariate and multivariate analyses were performed to identify factors associated with survival. In 102 evaluable patients, the median overall survival time was 14 months (range, 1 to > 41 months). Midtherapy CTC levels correlated with FDG-PET/CT response in 68 (67%) of 102 evaluable patients. In univariate analysis, midtherapy CTC counts and FDG-PET/CT response predicted overall survival (P < .001 and P = .001, respectively). FDG-PET/CT predicted overall survival (P = .0086) in 31 (91%) of 34 discordant patients who had fewer than five CTCs at midtherapy. Only midtherapy CTC levels remained significant in a multivariate analysis (P = .004). Detection of five or more CTCs during therapeutic monitoring can accurately predict prognosis in MBC beyond metabolic response. FDG-PET/CT deserves a role in patients who have fewer than five CTCs at midtherapy. Prospective trials should evaluate the most sensitive and cost-effective modality for therapeutic monitoring in MBC.


Somatic hypermutation of human mitochondrial and nuclear DNA by APOBEC3 cytidine deaminases, a pathway for DNA catabolism.
The human APOBEC3 (A3A-A3H) locus encodes six cytidine deaminases that edit single-stranded DNA, the result being DNA peppered with uridine. Although several cytidine deaminases are clearly restriction factors for retroviruses and hepadnaviruses, it is not known if APOBEC3 enzymes have roles outside of these settings. It is shown here that both human mitochondrial and nuclear DNA are vulnerable to somatic hypermutation by A3 deaminases, with APOBEC3A standing out among them. The degree of editing is much greater in patients lacking the uracil DNA-glycolyase gene, indicating that the observed levels of editing reflect a dynamic composed of A3 editing and DNA catabolism involving uracil DNA-glycolyase. Nonetheless, hyper- and lightly mutated sequences went hand in hand, raising the hypothesis that recurrent low-level mutation by APOBEC3A could catalyze the transition from a healthy to a cancer genome.


The need to reform personal injury law leaving scientific disputes to scientists.
Personal injury law is staggeringly inefficient as a system of victim compensation. There is little reason to assume that it importantly curtails unreasonably dangerous conduct, yet there is good reason to conclude that it promotes socially undesirable behavior. Moreover, the tort law system ill serves the goal of individual justice, in part because it assumes that lay juries can correctly decide complex scientific issues. Several methods of replacing tort law with other compensation systems are surveyed and a specific, balanced reform package is proposed.


Deciphering the molecular bases of Mycobacterium tuberculosis binding to the lectin DC-SIGN reveals an underestimated complexity.
Interactions between dendritic cells and Mycobacterium tuberculosis, the aetiological agent of tuberculosis in humans, are thought to be central to anti-mycobacterial immunity. We have previously shown that M. tuberculosis binds to human monocyte-derived dendritic cells mostly through the C-type lectin DC-SIGN (dendritic-cell-specific intercellular molecule-3-grabbing non-integrin)/CD209, and we have suggested that DC-SIGN may discriminate between mycobacterial species through recognition of the mannose-capping residues on the lipoglycan lipoarabinomannan of the bacterial envelope. Here, using a variety of fast- and slow-growing Mycobacterium species, we provide further evidence that mycobacteria recognition by DC-SIGN may be restricted to species of the M. tuberculosis complex. Fine analyses of the lipoarabinomannan molecules purified from these species show that the structure and amount of these molecules alone cannot account for such a preferential recognition. We propose that M. tuberculosis recognition by DC-SIGN relies on both a potential difference of accessibility of lipoarabinomannan in its envelope and, more probably, on the binding of additional ligands, possibly including lipomannan, mannose-capped arabinomannan, as well as the mannosylated 19 kDa and 45 kDa [Apa (alanine/proline-rich antigen)] glycoproteins. Altogether, our results reveal that the molecular basis of M. tuberculosis binding to DC-SIGN is more complicated than previously thought and provides further insight into the mechanisms of M. tuberculosis recognition by the immune system.


Sex-Related Differences in Plasma Oxytocin Levels in Humans.
Increasing evidence supports a key role of Oxytocin (OT) as a modulator of social relationships in mammals. The aim of the present study was to investigate possible sex-related differences in plasma OT levels in human beings. Forty-five healthy men and 45 women (mean age: 34.9 ± 6.2 years), were included in the study. Plasma preparation, peptide extraction and OT radioimmunoassay were carried out according to standardized methods. The results showed that OT plasma levels (pg / ml, mean ± SD) were significantly higher in women than in men (4.53 ± 1.18 vs 1.53 ± 1.19, p ˂ 0.001). The present finding demonstrates sex-related differences in plasma OT levels in humans. It is tempting to hypothesize that such differences might be related to behaviours, attitudes, as well as susceptibility to stress response, resilience and social emotions specific of women and men.


The ATAC acetyl transferase complex controls mitotic progression by targeting non-histone substrates.
All DNA-related processes rely on the degree of chromatin compaction. The highest level of chromatin condensation accompanies transition to mitosis, central for cell cycle progression. Covalent modifications of histones, mainly deacetylation, have been implicated in this transition, which also involves transcriptional repression. Here, we show that the Gcn5-containing histone acetyl transferase complex, Ada Two A containing (ATAC), controls mitotic progression through the regulation of the activity of non-histone targets. RNAi for the ATAC subunits Ada2a/Ada3 results in delayed M/G1 transition and pronounced cell division defects such as centrosome multiplication, defective spindle and midbody formation, generation of binucleated cells and hyperacetylation of histone H4K16 and alpha-tubulin. We show that ATAC localizes to the mitotic spindle and controls cell cycle progression through direct acetylation of Cyclin A/Cdk2. Our data describes a new pathway in which the ATAC complex controls Cyclin A/Cdk2 mitotic function: ATAC/Gcn5-mediated acetylation targets Cyclin A for degradation, which in turn regulates the SIRT2 deacetylase activity. Thus, we have uncovered an essential function for ATAC in regulating Cyclin A activity and consequent mitotic progression.


Heart rate-dependence of arterial distensibility in vivo.
Viscous and inertial components contribute to arterial distensibility and compliance in vitro. The purpose of our study was to determine whether this phenomenon is of relevance in vivo, namely, whether arterial compliance is altered by an increase in heart rate. Arterial diameter was assessed by an echo-Doppler device in a common carotid and femoral artery, namely, in a large elastic and a muscle artery. The studies were performed in 12-week-old pentobarbitone-anaesthetized Wistar-Kyoto rats subjected to atrial pacing via a transjugular unipolar catheter at five different randomly sequenced rates (280, 310, 340, 370 and 400 beats/min). After each stage, spontaneous sinus rhythm was allowed to return. Blood pressure was measured via a catheter inserted into the carotid or femoral artery contralateral to the vessels in which the diameter was measured. Arterial compliance and distensibility values were derived according to the Langewouters formula. A progressive increase in heart rate caused by pacing was accompanied by progressive and marked reductions in carotid artery compliance and distensibility. When quantified by the area under the distensibility-pressure or compliance-pressure curve the reduction was in the range 15-43%. Although a tendency to a similar phenomenon was observed in the femoral artery, in the latter vessel the reduction in distensibility and compliance was less marked and statistically insignificant. In the anaesthetized rat acute increases in heart rate are accompanied by reductions in arterial compliance and distensibility. The effect is greater in elastic than in muscle arteries.


"I can't imagine my life without it!" Curling and health promotion: a photovoice study.
The sport of curling is an important activity in rural Canada, with many female participants. The health of women in rural Canada is much compromised compared to that of their urban counterparts, yet little research has explored ways to promote the health of rural women. This study examined the influence of curling on the health of women in rural Canada. A sample of 15 women and girls aged 12 to 72 from 2 communities in southwestern Ontario took pictures, kept logbooks, and participated in 2 group sessions to discuss the influence of curling on their health. The findings reveal that curling facilitates social connections, enhances physical and mental health, and provides a valued and visible way to support rural life. Clearly, curling promotes the health and community life of rural women in significant ways. More support for curling in rural settings is needed, and additional research on the topic of curling and the health of rural women is indicated.


Is the adoption and safe families act influencing child welfare outcomes for families with substance abuse issues?
The Adoption and Safe Families Act (ASFA) was designed to promote more timely permanent placements for children in the child welfare system. To date, however, available data have said little about whether ASFA is meeting its intended goals. This study looks at the impact of ASFA on parents struggling with substance abuse issues. The authors compared child welfare outcomes, pre- and post-ASFA, for children of more than 1,900 substance-abusing women with some treatment involvement. After the implementation of ASFA, children in this study spent less time in foster care, were placed in permanent settings more quickly, and were more likely to be adopted than remain in long-term foster care. The proportion of children who were reunified with their parent or parents stayed the same. These outcomes were apparent even controlling for case and family characteristics. Results are discussed in terms of the influence of ASFA on service delivery systems.


Endoscope-assisted thoracoscopic technique for esophageal bronchogenic cyst which presented elevated CA125.
A 26-year-old male was admitted to our hospital with a complaint of mild dysphagia. Preoperative examinations revealed a cystic lesion at the lower esophagus, thought to be a bronchogenic cyst. The patient underwent successful thoracoscopic tumor extirpation, assisted by endoscopy, without undergoing major thoracotomy. Thoracoscopic surgery is recommended for this entity, but the muscle layer is often injured. Endoscopic ultrasonography was very useful, as it could detect damage to the proper muscle layer of the esophagus near the tumor. Thus, we could predict preoperatively that the defect in the muscle layer, which must be closed, would be caused by the surgery. Additionally, endoscopy easily facilitated the detection of the regional defect and the border of the proper muscle layer, and also could be used as a stent during wall suturing. This technique enabled us to avoid postoperative stenosis of the esophagus. Furthermore, our case presented a high level of CA125. The preoperative serum CA125 value was 93.8 U/ml and intratumoral CA125 concentration was 1,690,000 U/ml. Serum CA125 decreased to 16.5 U/ml after surgery. In this paper, we present a new strategy of the treatment for bronchogenic cyst and a review the literature.


Three-dimensional endomicroscopy of the human colon using optical coherence tomography.
Three-dimensional (3D) endomicroscopy imaging of the human gastrointestinal tract is demonstrated in vivo using a swept source optical coherence tomography (OCT) system. 3D datasets of normal and pathologic regions of the colon, rectum, and anal verge were obtained from seven volunteers undergoing diagnostic or therapeutic colonoscopy. 3D-OCT enables high resolution endomicroscopy examination through visualization of tissue architectural morphology using virtual cross-sectional images with arbitrary orientations as well as en face projection images. Axial image resolutions of 6 mum in tissue are obtained over a approximately 180 mm2 field with an imaging range of 1.6 mm. A Fourier domain mode locked (FDML) laser providing a tuning range of 180 nm at a sweep rate of 62 kHz is used as the system light source. This clinical pilot study demonstrates the potential of 3D-OCT for distinguishing normal from pathologic colorectal tissue, assessing endoscopic therapies and healing progression.


BCR-ABL1 expression, RT-qPCR and treatment decisions in chronic myeloid leukaemia.
RT-qPCR is used to quantify minimal residual disease (MRD) in chronic myeloid leukaemia (CML) in order to make decisions on treatment, but its results depend on the level of BCR-ABL1 expression as well as leukaemic cell number. The aims of the study were to quantify inter-individual differences in expression level, to determine the relationship between expression level and response to treatment, and to investigate the effect of expression level on interpretation of the RT-qPCR result. BCR-ABL1 expression was studied in 248 samples from 65 patients with CML by determining the difference between MRD quantified by RT-qPCR and DNA-qPCR. The results were analysed statistically and by simple indicative modelling. Inter-individual levels of expression approximated a normal distribution with an SD of 0.36 log. Expression at diagnosis correlated with expression during treatment. Response to treatment, as measured by the number of leukaemic cells after 3, 6 or 12 months of treatment, was not related to the level of expression. Indicative modelling suggested that interpretation of RT-qPCR results in relation to treatment guidelines could be affected by variation in expression when MRD was around 10% at 3 months and by both expression variation and Poisson variation when MRD was around or below the limit of detection of RT-qPCR. Variation between individuals in expression of BCR-ABL1 can materially affect interpretation of the RT-qPCR when this test is used to make decisions on treatment.


Latent and Abnormal Functional Connectivity Circuits in Autism Spectrum Disorder.
Autism spectrum disorder (ASD) is associated with disrupted brain networks. Neuroimaging techniques provide noninvasive methods of investigating abnormal connectivity patterns in ASD. In the present study, we compare functional connectivity networks in people with ASD with those in typical controls, using neuroimaging data from the Autism Brain Imaging Data Exchange (ABIDE) project. Specifically, we focus on the characteristics of intrinsic functional connectivity based on data collected by resting-state functional magnetic resonance imaging (rs-fMRI). Our aim was to identify disrupted brain connectivity patterns across all networks, instead of in individual edges, by using advanced statistical methods. Unlike many brain connectome studies, in which networks are prespecified before the edge connectivity in each network is compared between clinical groups, we detected the latent differentially expressed networks automatically. Our network-level analysis identified abnormal connectome networks that (i) included a high proportion of edges that were differentially expressed between people with ASD and typical controls; and (ii) showed highly-organized graph topology. These findings provide new insight into the study of the underlying neuropsychiatric mechanism of ASD.


Human perception of electrical stimulation on the surface of somatosensory cortex.
Recent advancement in electrocorticography (ECoG)-based brain-computer interface technology has sparked a new interest in providing somatosensory feedback using ECoG electrodes, i.e., cortical surface electrodes. We conducted a 28-day study of cortical surface stimulation in an individual with arm paralysis due to brachial plexus injury to examine the sensation produced by electrical stimulation of the somatosensory cortex. A high-density ECoG grid was implanted over the somatosensory and motor cortices. Stimulation through cortical surface electrodes over the somatosensory cortex successfully elicited arm and hand sensations in our participant with chronic paralysis. There were three key findings. First, the intensity of perceived sensation increased monotonically with both pulse amplitude and pulse frequency. Second, changing pulse width changed the type of sensation based on qualitative description provided by the human participant. Third, the participant could distinguish between stimulation applied to two neighboring cortical surface electrodes, 4.5 mm center-to-center distance, for three out of seven electrode pairs tested. Taken together, we found that it was possible to modulate sensation intensity, sensation type, and evoke sensations across a range of locations from the fingers to the upper arm using different stimulation electrodes even in an individual with chronic impairment of somatosensory function. These three features are essential to provide effective somatosensory feedback for neuroprosthetic applications.


Sleep changes during prophylactic treatment of migraine.
To assess sleep quality in patients with primary headaches before and after prophylactic treatment using a validated sleep-screening instrument. A total of 147 patients, including 63 tension type headache (TTH) and 84 migraine patients were included. Patients were examined in terms of frequency and severity of headaches and sleep quality before and 12 weeks after prophylactic treatment with either propranolol or amitriptyline. Baseline Visual Analogue Score (VAS) in migraine patients was 7.99 ± 1.39 compared with 6.86 ± 1.50 in TTH group (P < 0.001). VAS score after the first month of treatment was 6.08 ± 1.88 in migraine patients and 5.40 ± 1.61 in TTH (P = 0.023). VAS scores decreased after the third month of treatment to 4.32 ± 2.29 in migraine patients and 4.11 ± 1.66 in TTH patients (P = 0.344). The decrease was significant for patients treated with amitriptyline but not for those with propranolol. Baseline Pittsburgh Sleep Quality (PSQI) scores were 5.93 ± 2.43 in migraine patients and 6.71 ± 2.39 in TTH patients. Poor quality of sleep (PSQI ≥ 6) prior to prophylactic treatment was observed in 61.4% of migraine patients and in 77.7% of TTH patients. Comparison of PSQI scores before and 3 months following treatment showed significantly improved quality of sleep in all treatment groups; the greatest significance was detected in migraine patients with initial PSQI scores of ≥6 and treated with amitriptyline (P < 0.001). Increased understanding of routine objective sleep measures in migraine patients is needed to clarify the nature of sleep disturbances associated with primary headaches. This may in turn lead to improvements in headache treatments.


Reference values for lung function tests in men: regression equations with smoking variables.
Prediction formulas for static and dynamic spirometry, gas distribution, static lung mechanics and the transfer test were derived from measurements in healthy men. The measurements included total lung capacity, residual volume, airways resistance, static elastic recoil pressure of the lung, static compliance, closing volume, slope of the alveolar plateau (phase III), flow-volume variables (including mean transit time) during breathing of air or a helium/oxygen mixture, and conventional spirometric indices. The results from 146 smokers and 124 never-smokers were evaluated separately and combined. For all lung function tests a single regression equation was obtained. The prediction formulas included time-related smoking variables and were valid for both smokers and never-smokers. For many lung function tests, a nonlinear age coefficient resulted in a significant reduction in variance compared with simple linear models. Heavy tobacco smoking influenced most lung function tests less than ageing from 20 to 70 years, but for airways resistance, transfer factor and phase III the opposite was found.


Feasibility of resonant metalens for the subwavelength imaging using a single sensor in the far field.
This paper investigates the feasibility of the resonant metalens for the imaging beyond the diffraction limit using a single sensor in the far-field. It is shown that the resonant metalens can be related to the super-resonance phenomenon. We demonstrate that the super-resonance supports the enhancement of the information capacity of an imaging system, which is responsible for the subwavelength imaging of the probed objects by using a single sensor in combination with a broadband illumination. Such imaging concept has its unique advantage of producing real-time data when an object is illuminated by broadband waves, without the harsh requirements such as near-field scanning, mechanical scanning, or antenna arrays. The proposed method is expected to find its applications in nanolithography, detection, sensing, and subwavelength imaging in the near future.


Efficient conversion of chemical energy into mechanical work by Hsp70 chaperones.
Hsp70 molecular chaperones are abundant ATP-dependent nanomachines that actively reshape non-native, misfolded proteins and assist a wide variety of essential cellular processes. Here, we combine complementary theoretical approaches to elucidate the structural and thermodynamic details of the chaperone-induced expansion of a substrate protein, with a particular emphasis on the critical role played by ATP hydrolysis. We first determine the conformational free-energy cost of the substrate expansion due to the binding of multiple chaperones using coarse-grained molecular simulations. We then exploit this result to implement a non-equilibrium rate model which estimates the degree of expansion as a function of the free energy provided by ATP hydrolysis. Our results are in quantitative agreement with recent single-molecule FRET experiments and highlight the stark non-equilibrium nature of the process, showing that Hsp70s are optimized to effectively convert chemical energy into mechanical work close to physiological conditions.


Myeloid cell-derived inducible nitric oxide synthase suppresses M1 macrophage polarization.
Here we show that iNOS-deficient mice display enhanced classically activated M1 macrophage polarization without major effects on alternatively activated M2 macrophages. eNOS and nNOS mutant mice show comparable M1 macrophage polarization compared with wild-type control mice. Addition of N6-(1-iminoethyl)-L-lysine dihydrochloride, an iNOS inhibitor, significantly enhances M1 macrophage polarization while S-nitroso-N-acetylpenicillamine, a NO donor, suppresses M1 macrophage polarization. NO derived from iNOS mediates nitration of tyrosine residues in IRF5 protein, leading to the suppression of IRF5-targeted M1 macrophage signature gene activation. Computational analyses corroborate a circuit that fine-tunes the expression of IL-12 by iNOS in macrophages, potentially enabling versatile responses based on changing microenvironments. Finally, studies of an experimental model of endotoxin shock show that iNOS deficiency results in more severe inflammation with an enhanced M1 macrophage activation phenotype. These results suggest that NO derived from iNOS in activated macrophages suppresses M1 macrophage polarization.


Structural mechanism of demethylation and inactivation of protein phosphatase 2A.
Protein phosphatase 2A (PP2A) is an important serine/threonine phosphatase that plays a role in many biological processes. Reversible carboxyl methylation of the PP2A catalytic subunit is an essential regulatory mechanism for its function. Demethylation and negative regulation of PP2A is mediated by a PP2A-specific methylesterase PME-1, which is conserved from yeast to humans. However, the underlying mechanism of PME-1 function remains enigmatic. Here we report the crystal structures of PME-1 by itself and in complex with a PP2A heterodimeric core enzyme. The structures reveal that PME-1 directly binds to the active site of PP2A and that this interaction results in the activation of PME-1 by rearranging the catalytic triad into an active conformation. Strikingly, these interactions also lead to inactivation of PP2A by evicting the manganese ions that are required for the phosphatase activity of PP2A. These observations identify a dual role of PME-1 that regulates PP2A activation, methylation, and holoenzyme assembly in cells.


Patient perceptions of a personal health record: a test of the diffusion of innovation model.
Personal health records (PHRs) have emerged as an important tool with which patients can electronically communicate with their doctors and doctor's offices. However, there is a lack of theoretical and empirical research on how patients perceive the PHR and the differences in perceptions between users and non-users of the PHR. To apply a theoretical model, the diffusion of innovation model, to the study of PHRs and conduct an exploratory empirical study on the applicability of the model to the study of perceptions of PHRs. A secondary objective was to assess whether perceptions of PHRs predict the perceived value of the PHR for communicating with the doctor's office. We first developed a survey capturing perceptions of PHR use and other factors such as sociodemographic characteristics, access and use of technology, perceived innovativeness in the domain of information technology, and perceptions of privacy and security. We then conducted a cross-sectional survey (N = 1500). Patients were grouped into five groups of 300: PHR users (innovators, other users, and laggards), rejecters, and non-adopters. We applied univariate statistical analysis (Pearson chi-square and one-way ANOVA) to assess differences among groups and used multivariate statistical techniques (factor analysis and multiple regression analysis) to assess the presence of factors identified by the diffusion of innovation model and the predictors of our dependent variable (value of PHR for communicating with the doctor's office). Of the 1500 surveys, 760 surveys were returned for an overall response rate of 51%. Computer use among non-adopters (75%) was lower than that among PHR users (99%) and rejecters (92%) (P < .001). Non-adopters also reported a lower score on personal innovativeness in information technology (mean = 2.8) compared to 3.6 and 3.1, respectively, for users and rejecters (P < .001). Four factors identified by the diffusion of innovation model emerged in the factor analysis: ease of use, relative advantage, observability, and trialability. PHR users perceived greater ease of use and relative advantage of the PHR than rejecters and non-adopters (P < .001). Multiple regression analysis showed the following factors as significant positive predictors of the value of PHR for communicating with the doctor's office: relative advantage, ease of use, trialability, perceptions of privacy and security, age, and computer use. Our study found that the diffusion of innovation model fits the study of perceptions of the PHR and provides a suitable theoretical and empirical framework to identify the factors that distinguish PHR users from non-users. The ease of use and relative advantage offered by the PHR emerged as the most important domains among perceptions of PHR use and in predicting the value of the PHR. Efforts to improve uptake and use of PHRs should focus on strategies that enhance the ease of use of PHRs and that highlight the relative advantages of PHRs.


Alignment of Healthy Dietary Patterns and Environmental Sustainability: A Systematic Review.
To support food security for current and future generations, there is a need to understand the relation between sustainable diets and the health of a population. In recent years, a number of studies have investigated and compared different dietary patterns to better understand which foods and eating patterns have less of an environmental impact while meeting nutritional needs and promoting health. This systematic review (SR) of population-level dietary patterns and food sustainability extends and updates the SR that was conducted by the 2015 US Dietary Guidelines Advisory Committee, an expert committee commissioned by the federal government to inform dietary guidance as it relates to the committee's original conclusions. In the original SR, 15 studies met the criteria for inclusion; since then, an additional 8 studies have been identified and included. The relations between dietary intake patterns and both health and environmental outcomes were compared across studies, with methodologies that included modeling, life cycle assessment, and land use analysis. Across studies, consistent evidence indicated that a dietary pattern higher in plant-based foods (e.g., vegetables, fruits, legumes, seeds, nuts, whole grains) and lower in animal-based foods (especially red meat), as well as lower in total energy, is both healthier and associated with a lesser impact on the environment. This dietary pattern differs from current average consumption patterns in the United States. Our updated SR confirms and strengthens the conclusions of the original US Dietary Guidelines Advisory Committee SR, which found that adherence to several well-characterized dietary patterns, including vegetarian (with variations) diets, dietary guidelines-related diets, Mediterranean-style diets, the Dietary Approaches to Stop Hypertension (DASH) diet, and other sustainable diet scenarios, promotes greater health and has a less negative impact on the environment than current average dietary intakes.


Quality of Life among Persons with HIV/AIDS in Iran: Internal Reliability and Validity of an International Instrument and Associated Factors.
The purpose of this cross-sectional study on 191 HIV/AIDS patient was to prepare the first Persian translation of complete WHOQOL-HIV instrument, evaluate its reliability and validity, and apply it to determine quality of life and its associated factors in Tehran, Iran. Student's t-test was used to compare quality of life between groups. Mean Cronbach's α of facets in all six domains of instrument were more than 0.6 indicating good reliability. Item/total corrected correlations coefficients had a lower limit of more than 0.5 in all facets except for association between energy and fatigue facet and physical domain. Compared to younger participants, patients older than 35 years had significantly lower scores in overall quality of life (P = 0.003), social relationships (P = 0.021), and spirituality/religion/personal beliefs (P = 0.024). Unemployed patients had significantly lower scores in overall quality of life (P = 0.01), level of independence (P = 0.004), and environment (P = 0.001) compared to employed participants. This study demonstrated that the standard, complete WHOQOL-HIV 120 instrument translated into Farsi and evaluated among Iranian participants provides a reliable and valid basis for future research on quality of life for HIV and other patients in Iran.


Content and function of the self-definition in old and very old age.
Spontaneous self-definition was investigated in a heterogeneous sample of N = 516 participants of the Berlin Aging Study, aged between 70 and 103 years. The content of the self-definition revealed that old and very old persons view themselves as active and present-oriented. The self-definition also reflected an inward orientation, and central themes of life-review, health, and family. Participants generated more positive than negative evaluations in their self-definition, but the ratio of positive to negative evaluations was less favorable for the oldest old (> or = 85 years) than that of persons aged 70 to 84 years. Older individuals with more health-related constraints reported fewer and less rich self-defining domains (i.e., a less multifaceted self-definition). Positive emotional well-being was associated with naming more and richer self-defining domains. Multifacetness, however, did not buffer against the negative effect of low functional capacity on subjective well-being.


Enhanced detection of tuberculous mycobacteria in animal tissues using a semi-nested probe-based real-time PCR.
Bovine tuberculosis has been tackled for decades by costly eradication programs in most developed countries, involving the laboratory testing of tissue samples from allegedly infected animals for detection of Mycobacterium tuberculosis complex (MTC) members, namely Mycobacterium bovis. Definitive diagnosis is usually achieved by bacteriological culture, which may take up to 6-12 weeks, during which the suspect animal carcass and herd are under sanitary arrest. In this work, a user-friendly DNA extraction protocol adapted for tissues was coupled with an IS6110-targeted semi-nested duplex real-time PCR assay to enhance the direct detection of MTC bacteria in animal specimens, reducing the time to achieve a diagnosis and, thus, potentially limiting the herd restriction period. The duplex use of a novel β-actin gene targeted probe, with complementary targets in most mammals, allowed the assessment of amplification inhibitors in the tissue samples. The assay was evaluated with a group of 128 fresh tissue specimens collected from bovines, wild boars, deer and foxes. Mycobacterium bovis was cultured from 57 of these samples. Overall, the full test performance corresponds to a diagnostic sensitivity and specificity of 98.2% (CIP95% 89.4-99.9%) and 88.7% (CIP95% 78.5-94.7%), respectively. An observed kappa coefficient was estimated in 0.859 (CI P95% 0.771-0.948) for the overall agreement between the semi-nested PCR assay and the bacteriological culture. Considering only bovine samples (n = 69), the diagnostic sensitivity and specificity were estimated in 100% (CIP95% 84.0-100%) and 97.7% (CIP95% 86.2-99.9%), respectively. Eight negative culture samples exhibiting TB-like lesions were detected by the semi-nested real-time PCR, thus emphasizing the increased potential of this molecular approach to detect MTC-infected animal tissues. This novel IS6110-targeted assay allows the fast detection of tuberculous mycobacteria in animal specimens with very high sensitivity and specificity, being amenable and cost effective for use in the routine veterinary diagnostic laboratory with further automation possibilities.