Prognosis of histological cirrhosis in type 1 autoimmune hepatitis.
Cirrhosis connotes irreversible damage to the liver and shortened life expectancy. The aim of this study was to evaluate the impact of cirrhosis on treatment response and survival in type 1 autoimmune hepatitis. One hundred twenty-eight patients were evaluated for histological cirrhosis. Response to treatment, predictors for cirrhosis, and outcomes were determined. Thirty-seven patients (29%) had histological cirrhosis at entry, whereas 36 of the 91 patients without cirrhosis (40%) developed it during 39 +/- 32 months. Development of cirrhosis was predicted by lower serum albumin levels (P=0.04) and higher prothrombin times (P<0.001) at presentation. The frequencies of remission, relapse after drug withdrawal, and treatment failure were comparable in patients with and without cirrhosis at entry. The overall 10-year survival (93%) was similar to that of an age-and sex-matched cohort from the population at large (94%). Ten-year survival was not different between those with and those without cirrhosis at entry (89% and 90%, respectively), even with transplantation as an end point. Patients with histological cirrhosis at presentation respond to corticosteroid therapy as well as patients without cirrhosis. Low serum albumin levels and hypoprothrombinemia at presentation predict the development of cirrhosis. Histological cirrhosis does not diminish survival expectations.


The Mechanistic Understanding of RAD51 Defibrillation: A Critical Step in BRCA2-Mediated DNA Repair by Homologous Recombination.
The cytotoxic action of anticancer drugs can be potentiated by inhibiting DNA repair mechanisms. RAD51 is a crucial protein for genomic stability due to its critical role in the homologous recombination (HR) pathway. BRCA2 assists RAD51 fibrillation and defibrillation in the cytoplasm and nucleus and assists its nuclear transport. BRC4 is a peptide derived from the fourth BRC repeat of BRCA2, and it lacks the nuclear localization sequence. Here, we used BRC4 to (i) reverse RAD51 fibrillation; (ii) avoid the nuclear transport of RAD51; and (iii) inhibit HR and enhance the efficacy of chemotherapeutic treatments. Specifically, using static and dynamic light scattering, transmission electron microscopy, and microscale thermophoresis, we show that BRC4 eroded RAD51 fibrils from their termini through a "domino" mechanism and yielded monomeric RAD51 with a cumulative nanomolar affinity. Using cellular assays (BxPC-3, pancreatic cancer), we show that a myristoylated BRC4 (designed for a more efficient cell entry) abolished the formation of nuclear RAD51 foci. The present study provides a molecular description of RAD51 defibrillation, an essential step in BRCA2-mediated homologous recombination and DNA repair.


Case report: insertion of a covered stent for treatment of a popliteal artery pseudoaneurysm following total knee arthroplasty.
We report the use of a covered stent to successfully treat a pseudoaneurysm of the popliteal artery, which occurred following total knee arthroplasty. Percutaneous endovascular repair offers a safe alternative to surgical management of this condition.


Cognitive deficits among people with schizophrenia and prediabetes or diabetes.
Both type 2 diabetes mellitus (T2DM) and schizophrenia are known to be associated with cognitive deficits. The impact of the comorbidities of T2DM or prediabetes (PD) on cognition among people with schizophrenia has been poorly researched. We evaluated the cognitive functioning of patients with schizophrenia and PD or T2DM and compared them to patients with schizophrenia with normal blood sugar. We retrospectively collated data on cognition, fasting blood glucose (FBG), lipids and other selected demographic and clinical variables of 171 patients with schizophrenia and 16 patients with schizoaffective disorder who were admitted to an inpatient rehabilitation facility in Western Australia from 2011 to 2018. The Brief Assessment of Cognition in Schizophrenia (BACS) was used to evaluate cognitive functioning. Parametric and non-parametric analyses were used to examine the study's aims. Sixty-six percent of the patients had normal blood sugar, 25% had PD and 9% had T2DM. The BACS composite score revealed an increasing gradient of cognitive deficits, ranging from mild to severe, between the normal, PD and T2DM groups, respectively. The T2DM group had a significantly lower composite score compared with the PD (p = 0.026) and normal groups (p < 0.001). On the BACS subtests, the scores of T2DM and PD patients were similar except for the token motor task, in which the T2DM group had significantly lower scores (p < 0.001). The T2DM group also had lower scores on the subtests of BACS, except memory tests, compared with those with normal blood sugar. There was no significant difference in the composite and subtest cognitive scores between the PD and normal groups. Our study revealed more pronounced cognitive deficits among patients with schizophrenia and dysglycaemia, particularly those with T2DM, compared with those with schizophrenia with normal blood sugar.


Metal-ligand solution equilibria studied by electrospray ionization mass spectrometry: effect of instrumental parameters.
Electrospray ionization mass spectrometry (ESI-MS) is being increasingly employed in the study of metal-ligand equilibria in aqueous solution. In the present work, the ESI-MS spectral changes due to different settings of the following instrumental parameters are analyzed: the solution flow rate (F(S)), the nebulizer gas flow rate (F(G)), the sprayer potential (E), and the temperature of the entrance capillary (T). Twenty-eight spectra were obtained for each of six samples containing aluminum(III) and 2,3-dihydroxypyridine at various pH, in the absence or in the presence of a buffer and of sodium ions. Among the considered instrumental parameters, T produced the largest effects on the ionic intensities. F(S) and F(G) affected the ESI-MS spectra to a lower extent than T. In the investigated conditions E had the weakest effects on the spectra.The correlations observed between the ionic intensities and these instrumental parameters were interpreted considering the presence of three kinds of perturbations occurring in the ESI-MS ion source: formation of some dimers in the droplets, different transfer efficiencies from the droplets to the gas phase for different complexes (according to their surface activity), and subsequent partial thermal decomposition of the dimers and of one of the monomeric complexes in the gas phase. Our results show that the evaluation of the effects produced in the ESI-MS spectra by a change of instrumental parameters can allow to identify the perturbations occurring when metal-ligand solutions are studied by ESI-MS.


The interactive effects of Eimeria acervulina infection and phytase for broiler chicks.
An experiment was conducted to determine the interactive effects of Eimeria acervulina infection and phytase in male broiler chicks. Chicks were standardized from 0 to 4 d posthatching, and the assay period was 5 to 15 d. Treatments were replicated with 6 pens of 5 chicks each. The initial and final BW were 67 and 363 g. A corn-soybean meal diet formulated to provide 1.26% total Lys and 3,200 kcal of ME/kg was used, and it was adequate in all other nutrients except Ca and nonphytate P (NPP) when appropriate. The treatments were in a 2 x 2 x 2 factorial arrangement: adequate Ca and NPP (1.0% Ca and 0.45% NPP) or inadequate Ca and NPP (0.80% Ca and 0.25% NPP), 0 or 600 phytase units/kg of diet, and uninfected or infected with 400,000 E. acervulina oocysts on d 0, 3, and 6 of the experiment. Daily gain, average daily feed intake, and gain:feed (GF) were reduced (P < 0.01) by the coccidial infection and the reduction in Ca and NPP. Phytase addition increased (P < 0.02) average daily gain and average daily feed intake, regardless of the Ca and NPP contents of the diet or the presence of coccidiosis. The GF was increased by phytase but only in uninfected chicks (phytase x coccidiosis, P < 0.02). Toe and tibia ash percentages were decreased (P < 0.01) in chicks fed diets deficient in Ca and NPP, but tibia ash was decreased more by Ca and NPP in healthy chicks than in infected chicks (coccidiosis x Ca and NPP, P < 0.02). Phytase increased (P < 0.02) toe and tibia ash but only in diets deficient in Ca and NPP (phytase x Ca and NPP, P < 0.01). Phytase increased toe ash percentage of healthy chicks fed diets deficient in Ca and NPP, but it had less of an effect in infected chicks fed diets deficient in Ca and NPP (coccidiosis x Ca and NPP x phytase, P < 0.08). Also, phytase was less effective in increasing tibia ash percentage in coccidiosis-infected chicks than in uninfected chicks (phytase x coccidiosis, P < 0.02). These data indicate that phytase is effective in the presence of a coccidial infection, but based on GF and tibia ash percentage, it may not be as effective as in uninfected chicks. Also, phytase increased average daily gain and average daily feed intake in uninfected chicks fed diets formulated to be adequate (or in excess) in all nutrients for male broiler chicks.


External jugular venous catheterization with a Groshong catheter for central venous access.
The Groshong catheter (GC) is commonly used as a peripherally inserted central catheter (PICC), and the external jugular vein (EJV) is not a common route for central venous access (CVA). Therefore, external jugular venous catheterization (EJVC) using a GC is rare. However, our experience has shown that this procedure is safe and simple for CVA. To assess the utility of EJVC using the GC for CVA. CVA was done using a GC via the EJV with a single puncture employing the maximal barrier precaution technique. Between March 2005 and June 2007, we attempted to perform EJVC using a GC via the EJV for 331 patients. Among of these patients, 301 patients received successful catheterization (success rate: 301/331 = 90.9%). The total period of catheter insertion was 4601 days, and the mean insertion period was 15.1 days (range 2-147 days). The main complications of catheter insertion were malposition (8/301; 2.7%) and, oozing or hematoma formation (8/301; 2.7%). There were no severe complications such as nerve injury, pneumothorax or arterial bleeding. Our experience demonstrates that EJVC using the GC is acceptable for CVA.


Effects of haloperidol on behavioral responses induced by electrical stimulation of medial prefrontal cortex.
The effects of haloperidol on circling and spying behaviors induced by electrical stimulation of the medial prefrontal cortex (PFC) were evaluated. Male Wistar rats with an electrode implanted into the left medial PFC (B: 2.5 mm A, 0.6 mm L and 2.7 mm V) were electrically stimulated in a sequence of ten 30-sec trains separated by 30-sec intervals (60 Hz) in each session, and simultaneously observed in the open field. The animals with circling (CI) and spying (SP) behaviors were treated with intraperitoneal haloperidol (HAL ip, 5 mg/kg, N = 6) and saline (SAL ip, N = 7) or intracortical HAL (ic, 5 micrograms, N = 6) and SAL (ic, N = 9), 20 min before the session of electrical stimulation. HAL ic significantly decreased (P < 0.05) CI (mean frequency +/- SEM: 0.5 +/- 0.16) and nonsignificantly decreased SP behavior (0.6 +/- 0.17) compared to SAL ic (CI: 0.9 0.02, SP: 1 +/- 0). HAL ip full abolished these behaviors (P < 0.05) (CI: 0.02 +/- 0, SP: 1 +/- 0) compared to SAL ip (CI: 0.86 +/- 0.06, SP: 0:93 +/- 0.06). These results show that haloperidol, a dopaminergic antagonist and antipsychotic agent, interfered significantly with the expression of behaviors induced by electrical stimulation of the left medial PFC, suggesting that the induction of these behaviors may involve the dopaminergic neurotransmission.


Functional and structural differences between the prion protein from two alleles prnp(a) and prnp(b) of mouse.
The prion protein is a glycoprotein expressed by neurones and other cells. In its holo-form it binds copper and exhibits superoxide dismutase activity. Studies in mice have led to the description of two distinct alleles. Differences in these alleles are linked to long and short incubation times following infection with scrapie. We studied recombinant mouse protein corresponding to the products of either allele and two intermediates carrying single amino-acid residue substitutions. The different forms of the prion protein exhibited differences in superoxide dismutase (SOD) activity and conformation. Intermediates with single substitutions were less stable than either allelic product. The findings provide insight into the differences between the two alleles and might have consequences for understanding differences in susceptibility to prion disease.


3D Printed Magneto-Active Microfiber Scaffolds for Remote Stimulation and Guided Organization of 3D In Vitro Skeletal Muscle Models.
This work reports the rational design and fabrication of magneto-active microfiber meshes with controlled hexagonal microstructures via melt electrowriting (MEW) of a magnetized polycaprolactone-based composite. In situ iron oxide nanoparticle deposition on oxidized graphene yields homogeneously dispersed magnetic particles with sizes above 0.5 µm and low aspect ratio, preventing cellular internalization and toxicity. With these fillers, homogeneous magnetic composites with high magnetic content (up to 20 weight %) are obtained and processed in a solvent-free manner for the first time. MEW of magnetic composites enabled the creation of skeletal muscle-inspired design of hexagonal scaffolds with tunable fiber diameter, reconfigurable modularity, and zonal distribution of magneto-active and nonactive material, with elastic tensile deformability. External magnetic fields below 300 mT are sufficient to trigger out-of-plane reversible deformation. In vitro culture of C2C12 myoblasts on three-dimensional (3D) Matrigel/collagen/MEW scaffolds showed that microfibers guided the formation of 3D myotube architectures, and the presence of magnetic particles does not significantly affect viability or differentiation rates after 8 days. Centimeter-sized skeletal muscle constructs allowed for reversible, continued, and dynamic magneto-mechanical stimulation. Overall, these innovative microfiber scaffolds provide magnetically deformable platforms suitable for dynamic culture of skeletal muscle, offering potential for in vitro disease modeling.


Managing deliberate self-harm in young people: an evaluation of a training program developed for school welfare staff using a longitudinal research design.
Although deliberate self-harm is prevalent among young people, many who engage in deliberate self-harm receive sub-optimal care. Although schools are a well placed setting to support young people who engage in self-harm there are no specific training packages designed to assist school welfare staff to support these young people.The current study aimed to design, deliver and evaluate a training course specifically for school staff. The study employed a longitudinal design. Two hundred and thirteen people participated in the training and evaluation. A questionnaire was administered at baseline, immediately after the training and at 6-month follow-up in order to determine if the training led to improvements in confidence when working with young people who self-harm, perceived skill, knowledge of, and attitudes towards people who self harm. Prior to the course, the majority of participants demonstrated relatively high levels of confidence, perceived skill and knowledge of self-harm and endorsed relatively positive attitudes towards people who engage in self-harm. Despite this, significant improvements were observed in terms of increased confidence, increased perceptions of skill along with increased knowledge of deliberate self-harm. These improvements were sustained over the follow-up period. The results demonstrated that the provision of specifically designed training can help school welfare staff to feel better equipped to support young people who are engaging in deliberate self-harm.


Secondary structure of component 8c-1 of alpha-keratin. An analysis of the amino acid sequence.
The amino acid sequence of component 8c-1 from alpha-keratin was analysed by using secondary-structure prediction techniques, homology search methods, fast Fourier-transform techniques to detect regularities in the linear disposition of amino acids, interaction counts to assess possible modes of chain aggregation and assessment of hydrophilicity distribution. The analyses show the following. The molecule has two lengths of coiled-coil structure, each about 20 nm long, one from residues 56-202 with a discontinuity from about residue 91 to residue 101, and the other from residues 219-366 with discontinuities from about residue 238 to residue 245 and at about residue 306. The acidic and basic residues in the coiled-coil segment between residues 102 and 202 show a 9,4-residue structural period in their linear disposition, whereas between residues 246 and 366 a period of 9.9 residues is observed in the positioning of ionic residues. Acidic and basic residues are out of phase by 180 degrees. Similar repeats occur in corresponding regions of other intermediate-filament proteins. The overall mean values for the repeats are 9.55 residues in the N-terminal region and 9.85 residues in the C-terminal region. The regions at each end of the protein chain (residues 1-55 and 367-412) are not alpha-helical and contain many potential beta-bends. The regions specified in have a significant degree of homology mainly due to a semi-regular disposition of proline and half-cystine residues on a three-residue grid; this is especially apparent in the C-terminal segment, in which short (Pro-Cys-Xaa)n regions occur. The coiled-coil segments of component 8c-1 bear a striking similarity to corresponding segments of other intermediate-filament proteins as regards sequence homology, structural periodicity of ionic residues and secondary/tertiary-structure predictions. The assessments of the probabilities that these homologies occurred by chance indicate that there are two populations of keratin filament proteins. The non-coiled-coil regions at each end of the chain are less hydrophilic than the coiled-coil regions. Ionic interactions between the heptad regions of components 8c-1 and 7c from the microfibrils of alpha-keratin are optimized when a coiled-coil structure is formed with the heptad regions of the constituent chains both parallel and in register.


Bistable Mathematical Model of Neutrophil Migratory Patterns After LPS-Induced Epigenetic Reprogramming.
The highly controlled migration of neutrophils toward the site of an infection can be altered when they are trained with lipopolysaccharides (LPS), with high dose LPS enhancing neutrophil migratory pattern toward the bacterial derived source signal and super-low dose LPS inducing either migration toward an intermediary signal or dysregulation and oscillatory movement. Empirical studies that use microfluidic chemotaxis-chip devices with two opposing chemoattractants showed differential neutrophil migration after challenge with different LPS doses. The epigenetic alterations responsible for changes in neutrophil migratory behavior are unknown. We developed two mathematical models that evaluate the mechanistic interactions responsible for neutrophil migratory decision-making when exposed to competing chemoattractants and challenged with LPS. The first model, which considers the interactions between the receptor densities of two competing chemoattractants, their kinases, and LPS, displayed bistability between high and low ratios of primary to intermediary chemoattractant receptor densities. In particular, at equilibrium, we observe equal receptor densities for low LPS (< 15ng/mL); and dominance of receptors for the primary chemoattractant for high LPS (> 15ng/mL). The second model, which included additional interactions with an extracellular signal-regulated kinase in both phosphorylated and non-phosphorylated forms, has an additional dynamic outcome, oscillatory dynamics for both receptors, as seen in the data. In particular, it found equal receptor densities in the absence of oscillation for super-low and high LPS challenge (< 0.4 and 1.1 <LPS< 375 ng/mL); equal receptor densities with oscillatory receptor dynamics for super-low LPS (0.5 < LPS< 1.1ng/mL); and dominance of receptors for the primary chemoattractant for super-high LPS (>376 ng/mL). Predicting the mechanisms and the type of external LPS challenge responsible for neutrophils migration toward pro-inflammatory chemoattractants, migration toward pro-tolerant chemoattractants, or oscillatory movement is necessary knowledge in designing interventions against immune diseases, such as sepsis.


Pharmacodynamics OF TNF α inhibitors for the treatment of psoriasis.
Introduction: The treatment of psoriasis with conventional topical therapies and disease-modifying anti-rheumatic drugs (DMARDs) is often linked to unsatisfactory outcomes and the risk of serious adverse events. Over the last decades, research advances in understanding the role of tumor necrosis factor alpha (TNF α) and other cytokines in the pathogenesis of psoriasis have driven the introduction of biologic agents targeting specific immune mediators in everyday clinical practice. TNF α inhibitors are a consolidated treatment option for patients with moderate-to-severe disease with remarkable efficacy and a reassuring safety profile.Areas covered: The PubMed database was searched using combinations of the following keywords: psoriasis, TNF α inhibitors, biologic therapy, pharmacodynamics, adalimumab, etanercept, infliximab, certolizumab pegol, golimumab, adverse effects. The aim of this review is to describe the pharmacodynamic profile of anti-TNF α inhibitors, currently approved by the European Medicines Agency (EMA) for the treatment of psoriasis, focusing on related clinical implications, also in comparison to the new generation biological therapies targeting the interleukin 23/interleukin 17 axis.Expert opinion: Pharmacodynamics of TNF α inhibitors should be fully considered in planning patient's therapy strategies, especially in case of secondary failures, poor adherence to treatment, instable psoriasis, high risk of infection, pregnant or lactating women, metabolic comorbidities, coexistence of other immune-mediated inflammatory diseases.


Neonatal Intubation Practice and Outcomes: An International Registry Study.
Neonatal tracheal intubation is a critical but potentially dangerous procedure. We sought to characterize intubation practice and outcomes in the NICU and delivery room (DR) settings and to identify potentially modifiable factors to improve neonatal intubation safety. We developed the National Emergency Airway Registry for Neonates and collected standardized data for patients, providers, practices, and outcomes of neonatal intubation. Safety outcomes included adverse tracheal intubation-associated events (TIAEs) and severe oxygen desaturation (≥20% decline in oxygen saturation). We examined the relationship between intubation characteristics and adverse events with univariable tests and multivariable logistic regression. We captured 2009 NICU intubations and 598 DR intubations from 10 centers. Pediatric residents attempted 15% of NICU and 2% of DR intubations. In the NICU, the first attempt success rate was 49%, adverse TIAE rate was 18%, and severe desaturation rate was 48%. In the DR, 46% of intubations were successful on the first attempt, with 17% TIAE rate and 31% severe desaturation rate. Site-specific TIAE rates ranged from 9% to 50% (P < .001), and severe desaturation rates ranged from 29% to 69% (P = .001). Practices independently associated with reduced TIAEs in the NICU included video laryngoscope (adjusted odds ratio 0.46, 95% confidence interval 0.28-0.73) and paralytic premedication (adjusted odds ratio 0.38, 95% confidence interval 0.25-0.57). We implemented a novel multisite neonatal intubation registry and identified potentially modifiable factors associated with adverse events. Our results will inform future interventional studies to improve neonatal intubation safety.


Effect of warfarin on activated partial thromboplastin time in patients receiving heparin.
The activated partial thromboplastin time (APTT) is used to adjust heparin sodium dosage. However, warfarin sodium is often administered concomitantly with heparin and may also affect the APTT and, therefore, heparin dose. We performed a prospective cohort study to quantify the effect of warfarin on the APTT in patients who are being treated with heparin. Serial assays of APTT, international normalized ratio, heparin levels, and functional levels of prothrombin (factor II) and factors VII and X were performed in 24 patients with acute venous thromboembolism who were treated with concomitant continuous intravenous heparin and warfarin. The effects of warfarin, as expressed by international normalized ratio and coagulation factor levels, on APTT were determined. Warfarin markedly affected APTT; for each increase of 1.0 in the international normalized ratio, the APTT increased 16 seconds (95% confidence interval, 10-22 seconds). The effects of warfarin and heparin on APTT were additive. Consequently, warfarin markedly altered the relationship between APTT and heparin levels; of the 29 blood samples with supratherapeutic APTT, 13 had a therapeutic heparin level and 10 had a subtherapeutic heparin level. In patients receiving concomitant heparin and warfarin therapy, APTT reflects the combined effects of both drugs. Because of the marked effect of warfarin on the APTT, decreasing heparin dose in response to a high APTT frequently results in subtherapeutic heparin levels.


Cutting edge: persistence of increased mast cell numbers in tissues links dermatitis to enhanced airway disease in a mouse model of atopy.
The development of chronic allergic dermatitis in early life has been associated with increased onset and severity of allergic asthma later in life. However, the mechanisms linking these two diseases are poorly understood. In this study, we report that the development of oxazolone-induced chronic allergic dermatitis, in a mouse model, caused enhanced OVA-induced allergic asthma after the resolution of the former disease. Our findings show that oxazolone-induced dermatitis caused a marked increase in tissue mast cells, which persisted long after the resolution of this disease. Subsequent OVA sensitization and airway challenge of mice that had recovered from dermatitis resulted in increased allergic airway hyperreactivity. The findings demonstrate that the accumulation of mast cells during dermatitis has the detrimental effect of increasing allergic airway hypersensitivity. Importantly, our findings also show that exposure to a given allergen can modify the immune response to an unrelated allergen.


Human eosinophil cytotoxicity-enhancing factor. II. Multiple forms synthesized by U937 cells and their relationship to thioredoxin/adult T cell leukemia-derived factor.
Recently, our laboratory reported the purification and partial amino acid sequence of a 10-kDa eosinophil cytotoxicity-enhancing factor (ECEF) polypeptide from the U937 cell source. This cytokine enhanced human eosinophil antibody-dependent cytotoxic function by greater than 200% and was half-maximally effective at a concentration of approximately 1 ng/ml. In this study, we describe the conditions required for ECEF synthesis and the use of rabbit antibody raised to 10-kDa ECEF to investigate the existence of related polypeptide species. Unstimulated U937 cells released an immunoreactive 14-kDA species. Cells stimulated with 7.5 micrograms/ml of LPS also released a 13-kDa species. Cells stimulated with 400 ng/ml of PMA also synthesized a 10-kDa species (equivalent in size to the form we had purified). This 10-kDa species remained primarily cell associated, but detectable amounts were released into the supernatant by 48 h of culture. In washed cell pellets, the location of the 10-kDa species was found to be in the plasma membrane, externally oriented, as determined by FACS analysis, iodination with the membrane impermeable reagent 125I-sulfosuccinimidyl-3-(4-hydroxyphenyl) propionate, and by its removal with brief trypsin treatment. Partial amino acid sequence data suggested that the 14-, 13-, and 10-kDa species all share the same N-terminal. The 14- and 10-kDa ECEF species were recovered by electroelution from SDS-PAGE gels and tested for activity in the assay of eosinophil cytotoxic function. Because of the amino acid sequence similarities between the ECEF species and thioredoxin (TRX), rTRX (synthesized in Escherichia coli and purified) was also tested for activity. The 14-kDa ECEF and rTRX induced a slight, but consistent and statistically significant enhancement of eosinophil cytotoxic function. By comparison, lower doses of the 10-kDa ECEF induced a major increase in cytotoxic function. Thus the forms of ECEF differ in size, conditions required for synthesis, trafficking by the U937 cell after synthesis, and biologic activity. It is likely that these considerations bear on the involvement of ECEF in the pathophysiology of eosinophilia in vivo.


Ion-specific clustering of metal-amphiphile complexes in rare earth separations.
The nanoscale structure of a complex fluid can play a major role in the selective adsorption of ions at the nanometric interfaces, which is crucial in industrial and technological applications. Here we study the effect of anions and lanthanide ions on the nanoscale structure of a complex fluid formed by metal-amphiphile complexes, using small angle X-ray scattering. The nano- and mesoscale structures we observed can be directly connected to the preferential transfer of light (La and Nd) or heavy (Er and Lu) lanthanides into the complex fluid from an aqueous solution. While toluene-based complex fluids containing trioctylmethylammonium-nitrate (TOMA-nitrate) always show the same mesoscale hierarchical structure regardless of lanthanide loading and prefer light lanthanides, those containing TOMA-thiocyanate show an evolution of the mesoscale structure as a function of the lanthanide loading and prefer heavy lanthanides. The hierarchical structure indicates the presence of attractive interactions between ion-amphiphile aggregates, causing them to form clusters. A clustering model that accounts for the hard sphere repulsions and short-range attractions between the aggregates has been adapted to model the X-ray scattering results. The new model successfully describes the nanoscale structure and helps in understanding the mechanisms responsible for amphiphile assisted ion transport between immiscible liquids. Accordingly, our results imply different mechanisms of lanthanide transport depending on the anion present in the complex fluid and correspond with anion-dependent trends in rare earth separations.


Linking molecular physiology to ecological realities.
Although molecular physiology and ecology have drifted apart as a consequence of early separation in the questions posed and techniques used, there is a resurgence of interest in forging links between them. Here we explore the reasons for this renewed interest and provide four examples of how this is happening. Specifically, we examine links between molecular physiology and ecological realities in insect responses to thermal stress, vertebrate responses to anoxia, metabolic fuel use and torpor in mammals, and the recently developed "metabolic theory of ecology." Several novel insights are emerging from integrated approaches to these problems that might not have come forward from any single perspective on them. Nonetheless, prospects for linking molecular physiology and ecological realities are likely to remain poor if greater focus is not given to developing these links. Mostly, this is a consequence of the differing approaches and "languages" adopted by these fields. We discuss approaches by which the prospects for synthetic work might be improved.


Retinal detachment associated with macular hole surgery: characteristics, mechanism, and outcomes.
To study the frequency, features, and outcomes of retinal detachment (RD) occurring after macular hole surgery. Retrospective review of all cases of macular hole surgery. Cases with postoperative RD were identified for study. Outcome measures included baseline demographic and ocular characteristics, RD features, surgical procedure, macular hole status, and final visual and anatomic outcomes. Retinal detachment occurred in 8 (1.8%) of 438 eyes undergoing macular hole surgery. The rate of RD was 3.5% early in the course of the surgeon's experience (first 200 cases) and 0.4% later in the surgeon's experience (after 200 cases) (P = 0.026). Two of the eight eyes with RD had undergone previous macular hole surgery. All six primary cases occurred after repair of stage 3 macular holes, which had involved peeling of the posterior cortical face. The RD involved the inferior quadrants in seven eyes and the macula in two eyes. The pathogenic retinal break was in the equatorial region in four eyes, was anterior to the equator in two eyes, and remained undetermined in two eyes. The retina was attached and the macular hole was closed in all cases at the final follow-up examination (mean 30 months). The macular hole reopened 2 years following successful reattachment of the RD in one eye. Final visual acuity was > or =20/60 in four eyes and <20/200 in one eye. Retinal detachment after macular hole surgery is uncommon, and may be related to posterior cortical vitreous stripping. Early detection of RD minimizes adverse visual and anatomic outcomes.


CYP1A1 immunohistochemistry is highly specific for angiofibroma of soft tissue among morphological mimics.
Angiofibroma of soft tissue (AFST) is a benign, morphologically distinctive tumour type that harbours recurrent AHRR::NCOA2 fusions in 60-70% of cases and shows a non-specific immunophenotype, expressing EMA in roughly half of cases. The AHRR::NCOA2 fusion results in increased expression of cytochrome P450 1A1 (CYP1A1); a recent study demonstrated CYP1A1 immunohistochemistry (IHC) to be moderately sensitive and highly specific for AFST. In this study, we sought to validate these findings in a larger independent cohort of 30 AFST, as well as 215 morphological mimics, including 30 solitary fibrous tumours, 29 myxoid liposarcomas, 28 low-to-intermediate grade myxofibrosarcomas (MFS), 20 atypical spindle cell lipomatous tumours (ASCLT), 20 cellular angiofibromas, 10 cases each of spindle cell lipoma, neurofibroma, malignant peripheral nerve sheath tumour, superficial angiomyxoma, cellular myxoma, soft tissue perineurioma and deep fibrous histiocytoma, and nine cases each of low-grade fibromyxoid sarcoma and mammary-type myofibroblastoma. We found CYP1A1 IHC to be 70% sensitive for AFST, with granular cytoplasmic staining in 21 of 30 tumours, and 98% specific, with staining in only five morphological mimics: two deep fibrous histiocytomas, one MFS, one cellular angiofibroma and one ASCLT. These findings confirm that CYP1A1 is 70% sensitive, consistent with the prevalence of AHRR::NCOA2 fusions that up-regulate this protein, and that it is highly specific among morphological mimics.


Progenitor-derived human endothelial cells evade alloimmunity by CRISPR/Cas9-mediated complete ablation of MHC expression.
Tissue engineering may address organ shortages currently limiting clinical transplantation. Off-the-shelf engineered vascularized organs will likely use allogeneic endothelial cells (ECs) to construct microvessels required for graft perfusion. Vasculogenic ECs can be differentiated from committed progenitors (human endothelial colony-forming cells or HECFCs) without risk of mutation or teratoma formation associated with reprogrammed stem cells. Like other ECs, these cells can express both class I and class II major histocompatibility complex (MHC) molecules, bind donor-specific antibody (DSA), activate alloreactive T effector memory cells, and initiate rejection in the absence of donor leukocytes. CRISPR/Cas9-mediated dual ablation of β2-microglobulin and class II transactivator (CIITA) in HECFC-derived ECs eliminates both class I and II MHC expression while retaining EC functions and vasculogenic potential. Importantly, dually ablated ECs no longer bind human DSA or activate allogeneic CD4+ effector memory T cells and are resistant to killing by CD8+ alloreactive cytotoxic T lymphocytes in vitro and in vivo. Despite absent class I MHC molecules, these ECs do not activate or elicit cytotoxic activity from allogeneic natural killer cells. These data suggest that HECFC-derived ECs lacking MHC molecule expression can be utilized for engineering vascularized grafts that evade allorejection.


A randomized, prospective trial of operative treatments for hyperparathyroidism in patients with multiple endocrine neoplasia type 1.
Hyperparathyroidism (HPT) in multiple endocrine neoplasia (MEN) type 1 is associated with multiglandular parathyroid disease. Previous retrospective studies comparing subtotal parathyroidectomy (SP) and total parathyroidectomy with autotransplantation (TP/AT) have not established clearly better outcomes with either procedure. Patients were assigned randomly to either SP or TP/AT and data were collected prospectively. The rates of persistent HPT, recurrent HPT, and postoperative hypoparathyroidism were compared. The study cohort included 32 patients randomized to receive either SP or TP/AT (mean follow-up, 7.5 ± 5.7 years). The overall rate of recurrent HPT was 19% (6/32). Recurrent HPT occurred in 4 of 17 patients (24%) treated with SP and 2 of 15 patients (13%) treated with TP/AT (P = .66). Permanent hypoparathyroidism occurred in 3 of 32 patients (9%) overall. The rate of permanent hypoparathyroidism was 12% in the SP group (2/17) and 7% in the TP/AT group (1/15). A second operation was performed in 4 of 17 patients initially treated with SP (24%), compared with 1 of 15 patients undergoing TP/AT (7%; P = .34). This randomized trial of SP and TP/AT in patients with MEN 1 failed to show any difference in outcomes when comparing results of SP versus TP/AT. Both procedures are associated with acceptable results, but SP may have advantages in that is involves only 1 surgical incision and avoids an obligate period of transient postoperative hypoparathyroidism.


Brain stem oxidative stress and its associated signaling in the regulation of sympathetic vasomotor tone.
There is now compelling evidence from studies in humans and animals that overexcitation of the sympathetic nervous system plays an important role in the pathogenesis of cardiovascular diseases. An excellent example is neurogenic hypertension, in which central sympathetic overactivation is involved in the development, staging, and progression of the disease, and one of the underlying mechanisms involves oxidative stress in key brain stem sites that are engaged in the regulation of sympathetic vasomotor tone. Using the rostral ventrolateral medulla (RVLM) and nucleus tractus solitarii (NTS) as two illustrative brain stem neural substrates, this article provides an overview of the impact of reactive oxygen species and antioxidants on RVLM and NTS in the pathogenesis of neurogenic hypertension. This is followed by a discussion of the redox-sensitive signaling pathways, including several kinases, ion channels, and transcription factors that underpin the augmentation in sympathetic vasomotor tone. In addition, the emerging view that brain stem oxidative stress is also causally related to a reduction in sympathetic vasomotor tone and hypotension during brain stem death, methamphetamine intoxication, and temporal lobe status epilepticus will be presented, along with the causal contribution of the oxidant peroxynitrite formed by a reaction between nitric oxide synthase II (NOS II)-derived nitric oxide and superoxide. Also discussed as a reasonable future research direction is dissection of the cellular mechanisms and signaling cascades that may underlie the contributory role of nitric oxide generated by different NOS isoforms in the differential effects of oxidative stress in the RVLM or NTS on sympathetic vasomotor tone.


Surgical site infections and the surgical care improvement project (SCIP): evolution of national quality measures.
Surgical site infection (SSI) continues to be a common if often times preventable infection. Considerable patient morbidity and economic cost are the consequences. Review of the pertinent English-language literature. The National Surgical Infection Prevention (SIP) Project was an initiative sponsored jointly by the Centers for Medicare and Medicaid Services and the U.S. Centers for Disease Control and Prevention to decrease the incidence of SSI in major surgical procedures. Three performance measures were developed to improve the timing of antibiotic administration and the selection of the drug and to reduce needless postoperative administration of antibiotics. The success of SIP led to the Surgical Care Improvement Project (SCIP), which is expanding performance measures in the area of prevention of SSI, and also is undertaking preventive measures against postoperative venous thromboembolism and cardiac events. Federal legislation has required hospitals to report rates of compliance with many of these process measures. Surgeons must use all documented strategies to prevent SSIs.


DUART: durvalumab after radiotherapy in patients with unresectable, stage III NSCLC who are ineligible for chemotherapy.
Consolidation durvalumab is standard of care in patients with unresectable, stage III non-small-cell lung cancer (NSCLC) without disease progression following chemoradiotherapy (the 'PACIFIC regimen'). However, many patients with poor performance status, older age or comorbidities may be ineligible for chemotherapy due to expected high toxicity. These patients typically receive radiotherapy alone, with poor survival outcomes. Based on the PACIFIC trial data, and the strong biological rationale for combining radiotherapy with anti-programmed cell death ligand-1 therapy, durvalumab following radiotherapy could provide additional survival benefit versus radiotherapy alone. Here, we describe the DUART trial, a Phase II, open-label, single-arm study assessing the safety and tolerability of durvalumab following radiotherapy in patients with unresectable, stage III NSCLC who are ineligible for chemotherapy (ClinicalTrials.gov Identifier: NCT04249362).


The origins of African Plasmodium vivax; insights from mitochondrial genome sequencing.
Plasmodium vivax, the second most prevalent of the human malaria parasites, is estimated to affect 75 million people annually. It is very rare, however, in west and central Africa, due to the high prevalence of the Duffy negative phenotype in the human population. Due to its rarity in Africa, previous studies on the phylogeny of world-wide P. vivax have suffered from insufficient samples of African parasites. Here we compare the mitochondrial sequence diversity of parasites from Africa with those from other areas of the world, in order to investigate the origin of present-day African P. vivax. Mitochondrial genome sequencing revealed relatively little polymorphism within the African population compared to parasites from the rest of the world. This, combined with sequence similarity with parasites from India, suggests that the present day African P. vivax population in humans may have been introduced relatively recently from the Indian subcontinent. Haplotype network analysis also raises the possibility that parasites currently found in Africa and South America may be the closest extant relatives of the ancestors of the current world population. Lines of evidence are adduced that this ancestral population may be from an ancient stock of P. vivax in Africa.


The human weaning process.
The weaning process in relation to the nutritional adequacy of human lactation is reviewed. Current trends in breast-feeding, both in industrialized countries and the developing world, are considered, as well as associated changes in weaning practices. Consideration is also given to infant growth patterns in the Western world and how these are being influenced by current dietary practices. The nutritional requirements of babies during the first year of life are discussed, and recent concepts indicating that current estimates might be in excess of physiologic needs are introduced. These theories are of fundamental practical importance in terms of defining the length of time that exclusive breast-feeding remains adequate and determining when the introduction of complementary foods becomes necessary on nutritional grounds. It is emphasized that the weaning process is naturally a lengthy one, lasting many months rather than a few weeks, and that the provision of additional foods need not inhibit a mother's capacity for lactation as long as these feeds are administered in the correct fashion.


Integrin-substrate interactions underlying shear-induced inhibition of the inflammatory response of endothelial cells.
Conditioning of endothelial cells by shear stress suppresses their response to inflammatory cytokines. We questioned whether signalling through different integrin-matrix interactions, previously associated with the pathogenic effects of disturbed flow, supported the anti-inflammatory action of steady shear. Primary human endothelial cells were cultured on different substrates and exposed to shear stress (2.0Pa) for varying periods before stimulation with tumour necrosis factor-α (TNF). Shear-conditioning inhibited cytokine-induced recruitment of flowing neutrophils. However, the effect was similar for culture on collagen, laminin or fibronectin, even when seeding was reduced to 2 hours, and shear to 3 hours before TNF treatment (to minimise deposition of endothelial matrix). Nevertheless, in short- or longer-term cultures, reduction in expression of β(1)-integrin (but not β(3)-integrin) using siRNA essentially ablated the effect of shear-conditioning on neutrophil recruitment. Studies of focal adhesion kinase (FAK) phosphorylation, siRNA against FAK and a FAK-inhibitor (PF573228) indicated that FAK activity was an essential component downstream of β(1)-integrin. In addition, MAP-kinase p38 was phosphorylated downstream of FAK and also required for functional modification. Mechanotransduction through β(1)-integrins, FAK and p38 is required for anti-inflammatory effects of steady shear stress. Separation of the pathways which underlie pathological versus protective responses of different patterns of flow is required to enable therapeutic modification or mimicry, respectively.


Closure of the middle ear with special reference to the development of the tegmen tympani of the temporal bone.
Closure of the middle ear is believed to be closely related to the evolutionary development of the mammalian jaw. However, few comprehensive descriptions are available on fetal development. We examined paraffin-embedded specimens of 20 mid-term human fetuses at 8-25 weeks of ovulation age (crown-rump length or CRL, 38-220 mm). After 9 weeks, the tympanic bone and the squamous part of the temporal bone, each of which was cranial or caudal to Meckel's cartilage, grew to close the lateral part of the tympanosquamosal fissure. At the same time, the cartilaginous tegmen tympani appeared independently of the petrous part of the temporal bone and resulted in the petrosquamosal fissure. Subsequently, the medial part of the tympanosquamosal fissure was closed by the descent of a cartilaginous inferior process of the tegmen tympani. When Meckel's cartilage changed into the sphenomandibular ligament and the anterior ligament of the malleus, the inferior process of the tegmen tympani interposed between the tympanic bone and the squamous part of the temporal bone, forming the petrotympanic fissure for the chorda tympani nerve and the discomalleolar ligament. Therefore, we hypothesize that, in accordance with the regression of Meckel's cartilage, the rapidly growing temporomandibular joint provided mechanical stress that accelerated the growth and descent of the inferior process of the tegmen tympani via the discomalleolar ligament. The usual diagram showing bony fissures around the tegmen tympani may overestimate the role of the tympanic bone in the fetal middle-ear closure.


Transdermal delivery of phosphorodiamidate Morpholino oligomers across hairless mouse skin.
The skin is the largest organ in the body and an obvious route for both local and systemic drug delivery. Antisense oligomers have tremendous potential as therapeutic agents for numerous diseases. The objective of this study was to investigate the influence of vehicle on the transdermal delivery of several phosphorodiamidate Morpholino oligomers (PMOs) with different sizes, lengths, base compositions, sequences, and lipophilicities. Eleven different PMOs were synthesized complementary to biologically relevant gene targets and delivered across hairless mouse skin in vitro using vehicles composed of 95% propylene glycol, 5% linoleic acid (PG/LA), water, 50% water:50% PG/LA, and 75% water:25% PG/LA. The data suggest that size, sequence and guanine composition all influence transdermal penetration. There was an inverse linear relationship between size and penetration for a given sequence when the PG/LA formulation was used (r2 = 0.94), but this trend was not evident when the vehicle contained water. An oligomer targeted to the gene p53 had lower than expected transdermal penetration based on its size, but was shown to localize within the skin, demonstrating that sequence and thus target will impact transdermal delivery. The presence of G-quartets correlated with better PMO penetration from a water vehicle. Overall, the data suggest that some oligomers and vehicles would be better for transdermal delivery and others for topical applications.


Empathy in general practice-the gap between wishes and reality: comparing the views of patients and physicians.
Empathy is regarded by patients and general practitioners (GPs) as fundamental in patient-GP communication. Patients do not always experience empathy and GPs encounter circumstances which hamper applying it. To explore why receiving and offering empathy during the encounter in general practice does not always meet the wishes of both patients and GPs. A qualitative research method, based on focus group interviews with patients and in-depth interviews with GPs, was carried out. Within the research process, iterative data collection and analysis were applied. Both patients and GPs perceive a gap between what they wish for with regard to empathy, and what they actually encounter in general practice. Patients report on circumstances which hamper receiving empathy and GPs on circumstances offering it. Various obstacles were mentioned: (i) circumstances related to practice organization, (ii) circumstances related to patient-GP communication or connectedness, (iii) differences between the patient's and the GP's expectations, (iv) time pressure and its causes and (v) the GP's individual capability to offer empathy. When patients do not receive empathy from their GP or practice staff, they feel frustrated. This causes a gap between their expectations on the one hand and their actual experiences on the other. GPs generally want to incorporate empathy; the GP's private, professional and psychological well-being appears to be an important contributing factor in practicing empathy in daily practice. But they encounter various obstacles to offer this. It is up to GPs to take responsibility for showing practice members the importance of an appropriate empathical behaviour towards patients.


Recent Progress in 2D Metal-Organic Framework-Related Materials.
2D metal-organic frameworks-based (2D MOF-related) materials benefit from variable topological structures, plentiful open active sites, and high specific surface areas, demonstrating promising applications in gas storage, adsorption and separation, energy conversion, and other domains. In recent years, researchers have innovatively designed multiple strategies to avoid the adverse effects of conventional methods on the synthesis of high-quality 2D MOFs. This review focuses on the latest advances in creative synthesis techniques for 2D MOF-related materials from both the top-down and bottom-up perspectives. Subsequently, the strategies are categorized and summarized for synthesizing 2D MOF-related composites and their derivatives. Finally, the current challenges are highlighted faced by 2D MOF-related materials and some targeted recommendations are put forward to inspire researchers to investigate more effective synthesis methods.


Quantifying the length and variance of the eukaryotic cell cycle phases by a stochastic model and dual nucleoside pulse labelling.
A fundamental property of cell populations is their growth rate as well as the time needed for cell division and its variance. The eukaryotic cell cycle progresses in an ordered sequence through the phases G1, S, G2, and M, and is regulated by environmental cues and by intracellular checkpoints. Reflecting this regulatory complexity, the length of each phase varies considerably in different kinds of cells but also among genetically and morphologically indistinguishable cells. This article addresses the question of how to describe and quantify the mean and variance of the cell cycle phase lengths. A phase-resolved cell cycle model is introduced assuming that phase completion times are distributed as delayed exponential functions, capturing the observations that each realization of a cycle phase is variable in length and requires a minimal time. In this model, the total cell cycle length is distributed as a delayed hypoexponential function that closely reproduces empirical distributions. Analytic solutions are derived for the proportions of cells in each cycle phase in a population growing under balanced growth and under specific non-stationary conditions. These solutions are then adapted to describe conventional cell cycle kinetic assays based on pulse labelling with nucleoside analogs. The model fits well to data obtained with two distinct proliferating cell lines labelled with a single bromodeoxiuridine pulse. However, whereas mean lengths are precisely estimated for all phases, the respective variances remain uncertain. To overcome this limitation, a redesigned experimental protocol is derived and validated in silico. The novelty is the timing of two consecutive pulses with distinct nucleosides that enables accurate and precise estimation of both the mean and the variance of the length of all phases. The proposed methodology to quantify the phase length distributions gives results potentially equivalent to those obtained with modern phase-specific biosensor-based fluorescent imaging.


Bone mineral density during maintenance treatment with supraphysiological doses of levothyroxine in affective disorders: a longitudinal study.
This prospective study was designed to determine whether patients with prophylaxis-resistant affective disorders, receiving adjunctive maintenance therapy with supraphysiological doses of levothyroxine (L-T4), show evidence of accelerated bone loss compared to the reference population database. In 21 patients, bone mineral density (BMD) of the spine (lumbar vertebrae L1-L4) and femur (femoral neck, trochanter, and Ward's triangle) was measured by dual energy X-ray absorptiometry (DXA). BMD measurement was performed first after patients had been on thyroid-stimulating hormone (TSH)-suppressive therapy with L-T4 (mean dose=411 mcg/d) for an average of 16.4 months and again after 33.6 months of L-T4 (mean dose=416 mcg/d) therapy. There was no statistically significant difference between the actual percentage decline in bone mineral density and the expected percentage decline in any of the measured bone regions. In a stepwise linear regression analysis, age was identified as a predictor of percentage change in BMD. After controlling for age, the only other variable that showed a consistent trend was the dose of L-T4, with higher doses being positively correlated with the percentage decline of BMD. Relatively small sample size, no bone density assessment prior to treatment with L-T4, no patient control group with mood disorders who did not receive L-T4 treatment, and bone density follow-up intervals were variable. This study did not demonstrate evidence that long-term treatment of affectively ill patients with supraphysiological doses of L-T4 significantly accelerates loss of bone mineral density compared to the age-matched reference population. However, the decline of BMD in one individual patient underscores that caution is indicated and that regular assessment of BMD during longer-term supraphysiological thyroid hormone treatment is needed.


Automatic Localization of the Pons and Vermis on Fetal Brain MR Imaging Using a U-Net Deep Learning Model.
An MRI of the fetus can enhance the identification of perinatal developmental disorders, which improves the accuracy of ultrasound. Manual MRI measurements require training, time, and intra-variability concerns. Pediatric neuroradiologists are also in short supply. Our purpose was developing a deep learning model and pipeline for automatically identifying anatomic landmarks on the pons and vermis in fetal brain MR imaging and suggesting suitable images for measuring the pons and vermis. We retrospectively used 55 pregnant patients who underwent fetal brain MR imaging with a HASTE protocol. Pediatric neuroradiologists selected them for landmark annotation on sagittal single-shot T2-weighted images, and the clinically reliable method was used as the criterion standard for the measurement of the pons and vermis. A U-Net-based deep learning model was developed to automatically identify fetal brain anatomic landmarks, including the 2 anterior-posterior landmarks of the pons and 2 anterior-posterior and 2 superior-inferior landmarks of the vermis. Four-fold cross-validation was performed to test the accuracy of the model using randomly divided and sorted gestational age-divided data sets. A confidence score of model prediction was generated for each testing case. Overall, 85% of the testing results showed a ≥90% confidence, with a mean error of <2.22 mm, providing overall better estimation results with fewer errors and higher confidence scores. The anterior and posterior pons and anterior vermis showed better estimation (which means fewer errors in landmark localization) and accuracy and a higher confidence level than other landmarks. We also developed a graphic user interface for clinical use. This deep learning-facilitated pipeline practically shortens the time spent on selecting good-quality fetal brain images and performing anatomic measurements for radiologists.


Δ-9-Tetrahydrocannabinol and Cannabidiol produce dissociable effects on prefrontal cortical executive function and regulation of affective behaviors.
The use of cannabis for therapeutic and recreational purposes is growing exponentially. Nevertheless, substantial questions remain concerning the potential cognitive and affective side-effects associated with cannabis exposure. In particular, the effects of specific marijuana-derived phytocannabinoids on neural regions such as the prefrontal cortex (PFC) are of concern, given the role of the PFC in both executive cognitive function and affective processing. The main biologically active phytocannabinoids, ∆-9-tetrahydrocannabinol (THC) and cannabidiol (CBD), interact with multiple neurotransmitter systems important for these processes directly within the PFC. Considerable evidence has demonstrated that acute or chronic THC exposure may induce psychotomimetic effects, whereas CBD has been shown to produce potentially therapeutic effects for both psychosis and/or anxiety-related symptoms. Using an integrative combination of cognitive and affective behavioral pharmacological assays in rats, we report that acute intra-PFC infusions of THC produce anxiogenic effects while producing no impairments in executive function. In contrast, acute infusions of intra-PFC CBD impaired attentional set-shifting and spatial working memory, without interfering with anxiety or sociability behaviors. In contrast, intra-PFC CBD reversed the cognitive impairments induced by acute glutamatergic antagonism within the PFC, and blocked the anxiogenic properties of THC, suggesting that the therapeutic properties of CBD within the PFC may be present only during pathologically aberrant states within the PFC. Interestingly, the effects of PFC THC vs. CBD were found to be mediated through dissociable CB1 vs. 5-HT1A-dependent receptor signaling mechanisms, directly in the PFC.


Comparison of diagnostics techniques in an outbreak of infectious laryngotracheitis from meat chickens.
Various diagnostics techniques were compared for their ability to detect infectious laryngotracheitis (ILT) during an outbreak in chickens aged between 4 and 21 wk. Gross lesions ranged from excess mucus to accumulation of fibrinonecrotic exudate in the larynx and trachea. Syncytial cells with intranuclear inclusion bodies were found in sinus, conjunctiva, larynx, trachea, lung, and air sac. Virus isolation in chicken embryos was attempted in every case. Negative-stain electron microscopy detected herpesvirus in only 6% of the cases. Yet, isolation of ILT virus in the chorioallantoic membrane was presumed by histology in >20% of the samples and confirmed by fluorescent antibody (FA) in 35% of the embryos inoculated with conjunctivas or tracheas from affected birds. Overall, results from histology and FA tests were highly correlated. FA test has the advantage over histology of being diagnostically specific for ILT virus. Polymerase chain reaction was the most sensitive test and detected the viral DNA even in cases where histology and FA were negative. ILT virus DNA was quantified by real-time polymerase chain reaction (Re-Ti ILTV). Histologic and FA results from larynx and trachea were negative if the concentration of the viral DNA was < or =4 of log10. A viral DNA concentration higher than log10 4, as determined by Re-Ti ILTV, was required for clinical ILT to be manifested.


Altered diastolic function in asymptomatic infants of mothers with gestational diabetes.
Left and right ventricular filling was studied prospectively in 50 full-term (39.4 +/- 1.3 wk) asymptomatic newborns of mothers with gestational diabetes mellitus (GDM). Their data were compared with those of 80 asymptomatic full-term (39.8 +/- 1.2 wk) infants who served as control subjects. Infants were examined in the immediate newborn period (less than 48 h) and then again at 2-4 and 6-9 wk. Although mean weight, length, and gestational age did not differ, the mean +/- SD left ventricular dimensions during diastole (1.73 +/- 0.15 vs. 1.81 +/- 0.18 cm, P = 0.007) and systole (1.22 +/- 0.15 vs. 1.31 +/- 0.17 cm, P = 0.004) were significantly lower in infants of mothers with GDM compared with control infants. Diastolic measurements suggested a shift from the early diastolic filling of the ventricle to the later period of atrial systole in infants of mothers with GDM. A lower initial one-third area fraction and a higher peak flow velocity and velocity time integral during atrial systole were noted at the mitral valve in infants of mothers with GDM. These changes had resolved by 2-4 wk of age. The altered diastolic filling patterns in infants of mothers with GDM indicate poor myocardial relaxation and/or decreased passive compliance of the ventricular myocardium. These alterations were observed in asymptomatic infants in the absence of left ventricular or septal hypertrophy. If exposed to significant stress such as asphyxia or sepsis, the observed myocardial dysfunction could lead to higher morbidity in these infants.


[Preparation of asiaticoside and madecassoside from the extract of Centella asiatica (L.) Urb using preparative high performance liquid chromatography].
A high performance liquid chromatographic (HPLC) method was successfully used to separate asiaticoside and madecassoside from the extract of Centella asiatica (L.) Urb. The raw material containing 20% asiaticoside and 45% madecassoside was dissolved in methanol and used for the preparation of the pure compounds. The flow rate of the mobile phase, injection volume and detection wavelength were optimized. The optimum operation parameters were then selected as follows: a C18 column (50 mm x 200 mm, 5 microm) with methanol-water (60 : 40, v/v) as the mobile phase at a flow rate of 100 mL/min, the detection wavelength of 220 nm, and the injection volume of 1.5 mL at the concentration of 40 mg/mL. Asiaticoside and madecassoside the can be obtained in a 20 min runtime, and the purities of the products were detected by melting point, thin layer chromatography (TLC) and HPLC method. The results showed that the purities of products were above 98%. The effective and rapid method has been successfully applied and to the preparation of asiaticoside and madecassoside from Centella asiatica (L. ) Urb.


Specification of regions of DNA replication initiation during embryogenesis in the 65-kilobase DNApolalpha-dE2F locus of Drosophila melanogaster.
In the early stage of Drosophila embryogenesis, DNA replication initiates at unspecified sites in the chromosome. In contrast, DNA replication initiates in specified regions in cultured cells. We investigated when and where the initiation regions are specified during embryogenesis and compared them with those observed in cultured cells by two-dimensional gel methods. In the DNA polymerase alpha gene (DNApolalpha) locus, where an initiation region, oriDalpha, had been identified in cultured Kc cells, repression of origin activity in the coding region was detected after formation of cellular blastoderms, and the range of the initiation region had become confined by 5 h after fertilization. During this work we identified other initiation regions between oriDalpha and the Drosophila E2F gene (dE2F) downstream of DNApolalpha. At least four initiation regions showing replication bubbles were identified in the 65-kb DNApolalpha-dE2F locus in 5-h embryos, but only two were observed in Kc cells. These results suggest that the specification levels of origin usage in 5-h embryos are in the intermediate state compared to those in more differentiated cells. Further, we found a spatial correlation between the active promoter regions for dE2F and the active initiation zones of replication. In 5-h embryos, two known transcripts differing in their first exons were expressed, and two regions close to the respective promoter regions for both transcripts functioned as replication origins. In Kc cells, only one transcript was expressed and functional replication origins were observed only in the region including the promoter region for this transcript.


Patient selection for heart transplant: balancing risk.
Heart failure incidence continues to rise despite a relatively static number of available donor hearts. Selecting an appropriate heart transplant candidate requires evaluation of numerous factors to balance patient benefit while maximizing the utility of scarce donor hearts. Recent research has provided new insights into refining recipient risk assessment, providing additional tools to further define and balance risk when considering heart transplantation. Recent publications have developed models to assist in risk stratifying potential heart transplant recipients based on cardiac and noncardiac factors. These studies provide additional tools to assist clinicians in balancing individual risk and benefit of heart transplantation in the context of a limited donor organ supply. The primary goal of heart transplantation is to improve survival and maximize quality of life. To meet this goal, a careful assessment of patient-specific risks is essential. The optimal approach to patient selection relies on integrating recent prognostication models with a multifactorial assessment of established clinical characteristics, comorbidities and psychosocial factors.


Crouzons syndrome: a case report with review of literature.
Louis Edouard Octave Crouzon, a French neurologist, in 1912, described the hereditary syndrome of craniofacial synostosis in a mother and son. He described the triad as skull deformities, facial anamolies and exopthalmos now known as Crouzon syndrome (CS). CS accounts for about 4.8% of all cases of craniosynostosis. We report a case of CS in 4 year old girl with characteristic features of cranial deformity, maxillary hypoplasia, cleft palate and exopthalmos.


Development, antibiotic production, and ribosome assembly in Streptomyces venezuelae are impacted by RNase J and RNase III deletion.
RNA metabolism is a critical but frequently overlooked control element affecting virtually every cellular process in bacteria. RNA processing and degradation is mediated by a suite of ribonucleases having distinct cleavage and substrate specificity. Here, we probe the role of two ribonucleases (RNase III and RNase J) in the emerging model system Streptomyces venezuelae. We show that each enzyme makes a unique contribution to the growth and development of S. venezuelae and further affects the secondary metabolism and antibiotic production of this bacterium. We demonstrate a connection between the action of these ribonucleases and translation, with both enzymes being required for the formation of functional ribosomes. RNase III mutants in particular fail to properly process 23S rRNA, form fewer 70S ribosomes, and show reduced translational processivity. The loss of either RNase III or RNase J additionally led to the appearance of a new ribosomal species (the 100S ribosome dimer) during exponential growth and dramatically sensitized these mutants to a range of antibiotics.


Historical faunal exchange between the Pontocaspian Basin and North America.
Ecrobia is a genus of small brackish-water mud snails with an amphi-Atlantic distribution. Interestingly, the species occurring in the northwestern Atlantic, Ecrobia truncata, is more closely related to the Pontocaspian taxa, Ecrobia grimmi and Ecrobia maritima, than to the species occurring in the northeastern Atlantic and Mediterranean Sea. At least three colonization scenarios may account for this peculiar biogeographical pattern: (1) a recent human-mediated dispersal, (2) a historical transatlantic interchange, and (3) a historical transpolar interchange. To test these three scenarios, we used five operational criteria-time of species divergence, first appearance in the fossil record, dispersal limitation as well as environmental filtering and biotic interactions along the potential migration routes. Specifically, we inferred a time-calibrated molecular phylogeny for Ecrobia and reconstructed a paleogeographical map of the Arctic Ocean at 2.5 million years ago (Mya). Based on the five operational criteria, scenarios 1 and 2 can likely be rejected. In contrast, all criteria support scenario 3 (historical transpolar interchange). It is therefore suggested that a bird-mediated and/or ocean current-mediated faunal interchange via the Arctic Ocean occurred during the Late Pliocene or Early Pleistocene. This dispersal was likely facilitated by reduced distances between the Eurasian and North American/Greenland landmasses, marine introgressions, and/or a stepping-stone system of brackish-water habitats in northern Siberia, as well as a lack of competition along the migration route. As for the direction of dispersal, the scientific data presented are not conclusive. However, there is clearly more support for the scenario of dispersal from the Pontocaspian Basin to North America than vice versa. This is the first study providing evidence for a natural faunal exchange between the Pontocaspian Basin and North America via the Arctic Ocean.


Distributed Phase Oscillatory Excitation Efficiently Produces Attractors Using Spike-Timing-Dependent Plasticity.
The brain is thought to represent information in the form of activity in distributed groups of neurons known as attractors. We show here that in a randomly connected network of simulated spiking neurons, periodic stimulation of neurons with distributed phase offsets, along with standard spike-timing-dependent plasticity (STDP), efficiently creates distributed attractors. These attractors may have a consistent ordered firing pattern or become irregular, depending on the conditions. We also show that when two such attractors are stimulated in sequence, the same STDP mechanism can create a directed association between them, forming the basis of an associative network. We find that for an STDP time constant of 20 ms, the dependence of the efficiency of attractor creation on the driving frequency has a broad peak centered around 8 Hz. Upon restimulation, the attractors self-oscillate, but with an oscillation frequency that is higher than the driving frequency, ranging from 10 to 100 Hz.


Integrated assessment of behavioral and environmental risk factors for Lyme disease infection on Block Island, Rhode Island.
Peridomestic exposure to Borrelia burgdorferi-infected Ixodes scapularis nymphs is considered the dominant means of infection with black-legged tick-borne pathogens in the eastern United States. Population level studies have detected a positive association between the density of infected nymphs and Lyme disease incidence. At a finer spatial scale within endemic communities, studies have focused on individual level risk behaviors, without accounting for differences in peridomestic nymphal density. This study simultaneously assessed the influence of peridomestic tick exposure risk and human behavior risk factors for Lyme disease infection on Block Island, Rhode Island. Tick exposure risk on Block Island properties was estimated using remotely sensed landscape metrics that strongly correlated with tick density at the individual property level. Behavioral risk factors and Lyme disease serology were assessed using a longitudinal serosurvey study. Significant factors associated with Lyme disease positive serology included one or more self-reported previous Lyme disease episodes, wearing protective clothing during outdoor activities, the average number of hours spent daily in tick habitat, the subject's age and the density of shrub edges on the subject's property. The best fit multivariate model included previous Lyme diagnoses and age. The strength of this association with previous Lyme disease suggests that the same sector of the population tends to be repeatedly infected. The second best multivariate model included a combination of environmental and behavioral factors, namely hours spent in vegetation, subject's age, shrub edge density (increase risk) and wearing protective clothing (decrease risk). Our findings highlight the importance of concurrent evaluation of both environmental and behavioral factors to design interventions to reduce the risk of tick-borne infections.


Peptide-induced negative selection of thymocytes activates transcription of an NF-kappa B inhibitor.
Negative selection eliminates thymocytes bearing autoreactive T cell receptors (TCR) via an apoptotic mechanism. We have cloned an inhibitor of NF-kappa B, I kappa BNS, which is rapidly expressed upon TCR-triggered but not dexamethasone- or gamma irradiation-stimulated thymocyte death. The predicted protein contains seven ankyrin repeats and is homologous to I kappa B family members. In class I and class II MHC-restricted TCR transgenic mice, transcription of I kappa BNS is stimulated by peptides that trigger negative selection but not by those inducing positive selection (i.e., survival) or nonselecting peptides. I kappa BNS blocks transcription from NF-kappa B reporters, alters NF-kappa B electrophoretic mobility shifts, and interacts with NF-kappa B proteins in thymic nuclear lysates following TCR stimulation. Retroviral transduction of I kappa BNS in fetal thymic organ culture enhances TCR-triggered cell death consistent with its function in selection.


Critical role of the Src homology 2 (SH2) domain of neuronal SH2B1 in the regulation of body weight and glucose homeostasis in mice.
SH2B1 is an SH2 domain-containing adaptor protein that plays a key role in the regulation of energy and glucose metabolism in both rodents and humans. Genetic deletion of SH2B1 in mice results in obesity and type 2 diabetes. Single-nucleotide polymorphisms in the SH2B1 loci and chromosomal deletions of the SH2B1 loci associate with obesity and insulin resistance in humans. In cultured cells, SH2B1 promotes leptin and insulin signaling by binding via its SH2 domain to phosphorylated tyrosines in Janus kinase 2 and the insulin receptor, respectively. Here we generated three lines of mice to analyze the role of the SH2 domain of SH2B1 in the central nervous system. Transgenic mice expressing wild-type, SH2 domain-defective (R555E), or SH2 domain-alone (DeltaN503) forms of SH2B1 specifically in neurons were crossed with SH2B1 knockout mice to generate KO/SH2B1, KO/R555E, or KO/DeltaN503 compound mutant mice. R555E had a replacement of Arg(555) with Glu within the SH2 domain. DeltaN503 contained an intact SH2 domain but lacked amino acids 1-503. Neuron-specific expression of recombinant SH2B1, but not R555E or DeltaN503, corrected hyperphagia, obesity, glucose intolerance, and insulin resistance in SH2B1 null mice. Neuron-specific expression of R555E in wild-type mice promoted obesity and insulin resistance. These results indicate that in addition to the SH2 domain, N-terminal regions of neuronal SH2B1 are also required for the maintenance of normal body weight and glucose metabolism. Additionally, mutations in the SH2 domain of SH2B1 may increase the susceptibility to obesity and type 2 diabetes in a dominant-negative manner.


Nonredundancy in the dispersal network of a generalist tropical forest tree.
Plant species with generalized dispersal mutualisms are considered to be robust to local frugivore extinctions because of redundancy between dispersal agents. However, real redundancy can only occur if frugivores have similar foraging and ranging patterns and if fruit is a limiting resource. We evaluated the quantitative and qualitative contributions of seed dispersers for an endochorus mast-fruiting species, Prunus javanica (Rosaceae) in Khao Yai National Park, Thailand, to evaluate the potential redundancy of dispersers. Data were collected from tree watches, seed/fruit traps, and seed transects under and away from fruiting trees, feeding and seed deposition by gibbons (Hylobates lar), and evaluations of seed and first-year seedling survival. We identified three clusters of dispersers within the network. Most (>80%) frugivore species observed were small birds and squirrels that were not functional dispersers, dropping most seeds under or very near the tree crown, where seedling survival was ultimately nil. Monkeys (Macaca leonina) were low-quality, short-range dispersers, but they dispersed large numbers of seeds and were responsible for 67% of surviving first-year seedlings. Gibbons and Oriental Pied Hornbills (Anthracoceros albirostris) handled few fruits, but they provided the highest quality service by carrying most seeds away from the canopy to medium and long distances, respectively. Although there was overlap in the deposition patterns of the functional dispersers, they displayed complementary, rather than redundant, roles in seed dispersal. Satiation of all functional dispersers further limited their capacity to "replace" one another. Redundancy must be evaluated at the community level because each type of disperser may shift to different species in the non-masting years of P. javanica. Our results underscore the need for research on broader spatial and temporal scales, which combines studies of dispersal and plant recruitment, to better understand mechanisms that maintain network stability.


The coevolution of genes and genetic codes: Crick's frozen accident revisited.
The standard genetic code is the nearly universal system for the translation of genes into proteins. The code exhibits two salient structural characteristics: it possesses a distinct organization that makes it extremely robust to errors in replication and translation, and it is highly redundant. The origin of these properties has intrigued researchers since the code was first discovered. One suggestion, which is the subject of this review, is that the code's organization is the outcome of the coevolution of genes and genetic codes. In 1968, Francis Crick explored the possible implications of coevolution at different stages of code evolution. Although he argues that coevolution was likely to influence the evolution of the code, he concludes that it falls short of explaining the organization of the code we see today. The recent application of mathematical modeling to study the effects of errors on the course of coevolution, suggests a different conclusion. It shows that coevolution readily generates genetic codes that are highly redundant and similar in their error-correcting organization to the standard code. We review this recent work and suggest that further affirmation of the role of coevolution can be attained by investigating the extent to which the outcome of coevolution is robust to other influences that were present during the evolution of the code.


Deorphanization of GPRC6A: a promiscuous L-alpha-amino acid receptor with preference for basic amino acids.
One of the most important tasks of molecular pharmacology is the deorphanization of the large number of G-protein-coupled receptors with unidentified endogenous agonists. We recently reported the cloning and analysis of expression of a novel human family C G-protein-coupled receptor, termed hGPRC6A. To identify agonists at this orphan receptor, we faced the challenges of achieving surface expression in mammalian cell lines and establishing an appropriate functional assay. Generating a chimeric receptor construct, h6A/5.24, containing the ligand binding amino-terminal domain (ATD) of hGPRC6A with the signal transducing transmembrane and C terminus of the homologous goldfish 5.24 receptor allowed us to overcome these obstacles. Homology modeling of the hGPRC6A ATD based on the crystal structure of the metabotropic glutamate receptor subtype 1 predicted interaction with alpha-amino acids and was employed to rationally select potential ligands. Measurement of Ca2+-dependent chloride currents in Xenopus laevis oocytes facilitated the deorphanization of h6A/5.24 and identification of L-alpha-amino acids as agonists. The most active agonists were basic L-alpha-amino acids, L-Arg, L-Lys, and L-ornithine, suggesting that these may function as endogenous signaling molecules. Measurement of intracellular calcium in tsA cells expressing h6A/5.24 allowed determination of EC50 values, which confirmed the agonist preferences observed in oocytes. Cloning, cell surface expression and deorphanization of the mouse ortholog further reinforces the assignment of the agonist preferences of hGPRC6A. This study demonstrates the utility of a chimeric receptor approach in combination with molecular modeling, for elucidating agonist interaction with GPRC6A, a novel family C G-protein-coupled receptor.


Accelerating Tropicalization and the Transformation of Temperate Seagrass Meadows.
Climate-driven changes are altering production and functioning of biotic assemblages in terrestrial and aquatic environments. In temperate coastal waters, rising sea temperatures, warm water anomalies and poleward shifts in the distribution of tropical herbivores have had a detrimental effect on algal forests. We develop generalized scenarios of this form of tropicalization and its potential effects on the structure and functioning of globally significant and threatened seagrass ecosystems, through poleward shifts in tropical seagrasses and herbivores. Initially, we expect tropical herbivorous fishes to establish in temperate seagrass meadows, followed later by megafauna. Tropical seagrasses are likely to establish later, delayed by more limited dispersal abilities. Ultimately, food webs are likely to shift from primarily seagrass-detritus to more direct-consumption-based systems, thereby affecting a range of important ecosystem services that seagrasses provide, including their nursery habitat role for fishery species, carbon sequestration, and the provision of organic matter to other ecosystems in temperate regions.


Recent patent-based review on the role of three-dimensional printing technology in pharmaceutical and biomedical applications.
Three-dimensional printing (3DP) is emerging as an innovative manufacturing technology for biomedical and pharmaceutical applications, since the US FDA approval of Spritam as a 3D-printed drug. In the present review, we have highlighted the potential benefits of 3DP technology in healthcare, such as the ability to create patient-specific medical devices and implants, as well as the possibility of on-demand production of drugs and personalized dosage forms. We have further discussed future research to optimize 3DP processes and materials for pharmaceutical and biomedical applications. Cohesively, we have put forward the current state of active patents and applications related to 3DP technology in the healthcare and pharmaceutical industries including hearing aids, prostheses, medical devices and drug-delivery systems.


Orai1 and STIM1 move to the immunological synapse and are up-regulated during T cell activation.
For efficient development of an immune response, T lymphocytes require long-lasting calcium influx through calcium release-activated calcium (CRAC) channels and the formation of a stable immunological synapse (IS) with the antigen-presenting cell (APC). Recent RNAi screens have identified Stim and Orai in Drosophila cells, and their corresponding mammalian homologs STIM1 and Orai1 in T cells, as essential for CRAC channel activation. Here, we show that STIM1 and Orai1 are recruited to the immunological synapse between primary human T cells and autologous dendritic cells. Both STIM1 and Orai1 accumulated in the area of contact between either resting or super-antigen (SEB)-pretreated T cells and SEB-pulsed dendritic cells, where they were colocalized with T cell receptor (TCR) and costimulatory molecules. In addition, imaging of intracellular calcium signaling in T cells loaded with EGTA revealed significantly higher Ca2+ concentration near the interface, indicating Ca2+ influx localized at the T cell/dendritic cell contact area. Expression of a dominant-negative Orai1 mutant blocked T cell Ca2+ signaling but did not interfere with the initial accumulation of STIM1, Orai1, and CD3 in the contact zone. In activated T cell blasts, mRNA expression for endogenous STIM1 and all three human homologs of Orai was up-regulated, accompanied by a marked increase in Ca2+ influx through CRAC channels. These results imply a positive feedback loop in which an initial TCR signal favors up-regulation of STIM1 and Orai proteins that would augment Ca2+ signaling during subsequent antigen encounter.


Mucosal melanoma: correlation of clinicopathologic, prognostic, and molecular features.
Although the presence of the t(12;22)(q13;q12) translocation (the defining molecular feature of malignant melanoma of soft parts/clear cell sarcoma) in cutaneous melanoma has been investigated, no large-scale studies have been performed among mucosal melanoma (MucM). In this study we assessed the prevalence of the EWSR1 rearrangement in primary MucM, and analyzed gross and microscopic features with their potential impact on diagnosis and prognosis. Overall, 132 specimens from 84 patients were included. A total of 55 cases had an intramucosal component. Survival of MucMs of the head and neck was associated with two independent factors: size and histology. Tumors more than 3 cm in greatest dimension had an average survival of 12.75 months; those 3 cm or less had an average survival of 38.3 months (P=0.035). Purely epithelioid tumors had an average worse survival of 16.8 months (P=0.028). A cut-off value of 1 mm for Breslow depth provided a statistically significant difference in survival at both 3 and 5 years (P=-0.02) by multivariate analysis in the gynecologic tract. At the molecular level three cases had a EWSR1 rearrangement by fluorescent in-situ hybridization, but only one with an intramucosal component. None of the 58 cases tested by PCR showed the presence of the EWSR1 rearrangement. With the exception of vulvar melanomas, the prognosis of mucosal-associated melanomas was poor and there was a suggestion that spindle morphology may be more favorable. Our study also showed that the EWSR1 rearrangement was very uncommon among MucM. Though 'clear cell sarcoma' is embedded in the sarcoma literature, the synonym 'melanoma of soft parts' has considerable justification in light of our evolving understanding of the molecular genetics in the family of malignant melanomas.


Plasma homocysteine, hypertension incidence, and blood pressure tracking: the Framingham Heart Study.
Plasma homocysteine is cross-sectionally associated with blood pressure in large, community-based studies. It is unknown whether elevated plasma homocysteine predicts hypertension incidence. We investigated the relations of baseline plasma total homocysteine levels to hypertension incidence and blood pressure tracking in 2104 Framingham Heart Study participants (mean age, 57 years; 58% women), who were free of hypertension, myocardial infarction, heart failure, atrial fibrillation, or renal failure at baseline. Baseline mean+/-SD plasma homocysteine was 10.1+/-3.7 micromol/L. On follow-up 4 years from baseline, 360 persons (17.1%) had developed hypertension, and 878 persons (41.7%) had progressed to a higher blood pressure stage. In unadjusted analyses, a 1-SD higher log homocysteine value was associated with increased odds of developing hypertension (odds ratio [OR], 1.18; 95% confidence interval [CI], 1.05 to 1.32) and increased odds of blood pressure progression (OR, 1.17; 95% CI, 1.07 to 1.27). The relations of plasma homocysteine to the incidence of hypertension or blood pressure progression were statistically nonsignificant in age- and sex-adjusted logistic regression models (OR, 0.98; 95% CI, 0.87 to 1.11 and OR, 1.05; 95% CI, 0.96 to 1.16, respectively) and in multivariable models adjusted for age, sex, body mass index, diabetes, interim weight change, smoking, serum creatinine, baseline blood pressure, and blood pressure category (OR, 0.92; 95% CI, 0.81 to 1.06 and OR, 1.07; 95% CI, 0.97 to 1.18, respectively). In conclusion, we found no major relation of baseline plasma homocysteine levels to hypertension incidence or longitudinal blood pressure progression in a large, community-based cohort of nonhypertensive individuals after adjustment for age, sex, and other important covariates.


Effects of Risk-Based Firearm Seizure Laws in Connecticut and Indiana on Suicide Rates, 1981-2015.
This study evaluated whether risk-based firearm seizure laws in Connecticut and Indiana affect suicide rates. A quasi-experimental design using annual state-level panel data from the 50 states between 1981 and 2015 was used. When analyses controlled for a range of risk factors for population-level suicide rates, the effects of Connecticut and Indiana's firearm seizure laws on firearm and nonfirearm suicide rates were evaluated by using the synthetic-control methodology and difference-in-place placebo tests. Sensitivity analyses employed regression-based difference-in-differences analyses with randomization inference. Indiana's firearm seizure law was associated with a 7.5% reduction in firearm suicides in the ten years following its enactment, an effect specific to suicides with firearms and larger than that seen in any comparison state by chance alone. Enactment of Connecticut's law was associated with a 1.6% reduction in firearm suicides immediately after its passage and a 13.7% reduction in firearm suicides in the post-Virginia Tech period, when enforcement of the law substantially increased. Regression-based sensitivity analyses showed that these findings were robust to alternative specifications. Whereas Indiana demonstrated an aggregate decrease in suicides, Connecticut's estimated reduction in firearm suicides was offset by increased nonfirearm suicides. Risk-based firearm seizure laws were associated with reduced population-level firearm suicide rates, and evidence for a replacement effect was mixed.


Sinus mucocele secondary to craniofacial trauma in a dog.
A sinsus mucocele formed in the frontal sinus of a dog secondary to obstruction of the nasofrontal opening. The dog was successfully treated by surgical enlargement of the nasofrontal opening, curettage of the sinus epithelium, and placement of a fat graft and drain in the sinus. Although mucocele formation is uncommon in animals, it should be considered as a differential diagnosis for masses of the sinonasal area. Characteristic radiographic and cytologic findings should alert the clinician to this diagnosis, and along with aggressive treatment, should shorten the diagnostic and treatment delay experienced by the dog in this report.


Appropriateness ratings of everyday behaviors in the United States now and 50 years ago.
A crucial aspect of social norms pertains to determining which behaviors are considered appropriate. Here we consider everyday behaviors. Some everyday behaviors are rated as more appropriate than others, and ratings of the appropriateness of a given behavior may vary over time. The objective of this study is to elucidate the reasons behind variation in appropriateness ratings of everyday behaviors in the United States. Our theory focuses on how the evaluation of the appropriateness of a behavior is influenced by its potential for externalities and internalities, and how this influence may cause a change in norms over time. Employing a preregistered design, we asked American participants to rate 37 different everyday behaviors based on their appropriateness in a range of common situations, as well as their potential negative externalities (e.g., being loud, being aggressive, taking up space) and positive internalities (e.g., pleasurability). Changes over time were calculated as the difference between mean ratings obtained in this study and ratings of the same behavior in a similar study conducted 50 years ago. As expected, overall appropriateness ratings of everyday behaviors are associated both with their externalities and their internalities, so that the least appropriate behaviors tend to have considerable potential for negative externalities and little potential for positive internalities. Moreover, behaviors that have considerable potential for negative externalities are perceived as less appropriate now than 50 years ago. By describing how social norms for everyday behaviors depend on the externalities and internalities of behaviors, this study contributes to theories about the emergence and change of social norms.


Attitudes of relatives of mucopolysaccharidosis type III patients toward preconception expanded carrier screening.
Preconception expanded carrier screening (ECS) aims to detect carrier couples of autosomal recessive (AR) disorders before pregnancy in order to increase reproductive autonomy of prospective parents. Genetic knowledge and knowledge gained from experience influence decision making on participation in genetic testing and understanding carrier test results. In this study we assessed whether parents and relatives of patients with the severe AR condition mucopolysaccharidosis type III (MPS III), who are expected to have genetic and experiential knowledge, have more positive attitudes toward ECS than the Dutch reference group. Parents of all MPS III patients known to the Dutch expert center were invited to participate and asked to invite first and second degree relatives. The online questionnaire started with an educational text, and assessed attitudes toward ECS, genetic knowledge and perceived MPS III severity. Results were compared with the Dutch population. Parents and relatives of MPS III patients (n = 159) scored higher on the genetic knowledge test and perceived MPS III as more severe compared with the general Dutch population (n = 781). Parents and relatives reported to be more likely to participate in ECS (84.3% and 62.5%, respectively) compared with the public (31%) (p < 0.001). Being a relative of a MPS III patient was the strongest variable in the regression analyses for intended ECS participation. Our results show that genetic knowledge influences ECS decision making. Therefore, appropriate information on ECS and genetic counseling is needed to enable prospective parents from the general population, including relatives of patients with severe hereditary disorders, to make informed decisions.


Kindling-induced potentiation in the piriform cortex.
At intensities sufficient to induce epileptiform afterdischarges, repeated electrical stimulation of limbic structures can lead to the development of permanent increases in the strength of the epileptiform response (kindling). Field potentials evoked by pulse stimulation are also increased in amplitude in a number of forebrain pathways following kindling. This kindling-induced potentiation effect is similar in many respects to the 'long-term potentiation' (LTP) effect which is produced by non-epileptogenic stimulation. There are, however, some interesting differences. For example, kindling-induced potentiation can far outlast LTP. In these experiments, we attempted to determine the longevity of the kindling-induced potentiation of the response evoked in the piriform cortex by olfactory bulb stimulation, following olfactory bulb kindling. This system was targeted because both the olfactory bulb and the piriform cortex are highly reactive kindling sites. In addition, we used the paired pulse technique to monitor facilitation and inhibition in this system. Kindling was found to induce a potentiation in the piriform field potential that lasted for at least 3 months (the period of the experiment) with little or no decay. Kindling also produced a decrease in paired pulse facilitation. In some animals the net facilitation was changed to a net depression. These results are consistent with the interpretation that kindling produces an increase in recurrent inhibition in the piriform cortex. The paired pulse measures, however, returned to near baseline levels over the 3-month test period.