National study of master teachers in deaf education: implications for teacher education.
After an extensive review of relevant literature, the investigators developed a questionnaire on teacher characteristics and behaviors in relation to master teachers and distributed it electronically to deaf education professionals. The questionnaire recipients represented administrators, experienced teachers (e.g., those with 3 or more years' experience), and college and university faculty responsible for preparing teachers of the deaf. Follow-up interactive interviews were conducted with representatives of each constituency. Considerable similarities were noted in the priorities assigned across the three different constituencies, including a value placed on strong communication skills, having a passion about teaching, being collaborative, remaining current in the field, helping students become independent learners, and employing cognitive strategies. Analyses of responses by constituency are provided, along with recommendations for action, particularly for teacher education programs that are intent on graduating more teachers with master teacher potential.


The incidence of adverse events in an Italian acute care hospital: findings of a two-stage method in a retrospective cohort study.
The promotion of safer healthcare interventions in hospitals is a relevant public health topic. This study is aimed to investigate predictors of Adverse Events (AEs) taking into consideration the Charlson Index in order to control for confounding biases related to comorbidity. The study was a retrospective cohort study based on a two-stage assessment tool which was used to identify AEs. In stage 1, two physicians reviewed a random sample of patient records from 2008 discharges. In stage 2, reviewers independently assessed each screened record to confirm the presence of AEs. A univariable and multivariable analysis was conducted to identify prognostic factors of AEs; socio-demographic and some main organizational variables were taken into consideration. Charlson comorbidity Index was calculated using the algorithm developed by Quan et al. A total of 1501 records were reviewed; mean patients age was 60 (SD: 19) and 1415 (94.3%) patients were Italian. Forty-six (3.3%) AEs were registered; they most took place in medical wards (33, 71.7%), followed by surgical ones (9, 19.6%) and intensive care unit (ICU) (4, 8.7%). According to the logistic regression model and controlling for Charlson Index, the following variables were associated to AEs: type of admission (emergency vs elective: OR 3.47, 95% CI: 1.60-7.53), discharge ward (surgical and ICU vs medical wards: OR 2.29, 95% CI: 1.00-5.21 and OR 4.80, 95% CI: 1.47-15.66 respectively) and length of stay (OR 1.03, 95% CI 1.01-1.04). Among patients experiencing AEs a higher frequency of elderly (≥65 years) was shown (58.7% vs 49.3% among patients without AEs) but this difference was not statistically significant. Interestingly, a higher percentage of patients admitted through emergency department was found among patients experiencing AEs (69.7% vs 55.1% among patients without AEs). The incidence of AEs was associated with length of stay, type of admission and unit of discharge, independently by comorbidity. On the basis of our results, it appears that organizational characteristics, taking into account the adjustment for comorbidity, are the main factors responsible for AEs while patient vulnerability played a minor role.


Combined therapy of deoxyspergualin and plasmapheresis: a useful treatment for antibody-mediated acute rejection after kidney transplantation.
Antibody-mediated acute rejection (AbAR) is one of the primary causes of graft impairment in kidney transplant recipients. Deoxyspergualin (DSG), which displays an antiproliferative action against antigen-stimulated B cells inhibiting antibody production, may be effective to rescue AbAR in combination with plasmapheresis by suppressing antibody production and elimination. In the present study, we report our experience with DSG/plasmapheresis therapy for the treatment of AbAR. Five kidney transplant patients experienced a steroid-resistant acute rejection requiring dialysis followed by an AbAR that was confirmed by biopsy and flow cytometry crossmatch (FCXM) results. DSG was administration at 3 mg/kg per day for 10 days with plasmapheresis reduce antidonor antibody. Treatment outcome, effectiveness, and adverse events were examined; in two cases sequential FCXM examinations were performed to evaluate antibody status. All five patients received DSG/plasmapheresis therapy. The number of plasmapheresis treatments ranged from 1 to 9 according to treatment outcomes. Four patients recovered graft function following treatment; whereas one showed no response to the treatment, and the graft was lost. No serious side effects or infections were observed during or after treatment. Monitoring of sequential FCXM correlated with the clinical course. AbAR shows a worse prognosis than cellular rejection. It is refractory to conventional antirejection therapy. In the present study, DSG/plasmapheresis therapy was effective in four of five patients (80%) with AbAR. It may be considered the first choice of treatment for cases of acute humoral rejection.


Prediction of genetic growth curves in pigs.
Genetic growth curves of boars in a test station were predicted on daily weight records collected by automated weighing scales. The data contained 121 865 observations from 1477 Norwegian Landrace boars and 108 589 observations from 1300 Norwegian Duroc boars. Random regression models using Legendre polynomials up to second order for weight at different ages were compared for best predicting ability and Bayesian information criterion (BIC) for both breeds. The model with second-order polynomials had best predictive ability and BIC. The heritability for weight, based on this model, was found to vary along the growth trajectory between 0.32-0.35 for Duroc and 0.17-0.25 for Landrace. By varying test length possibility to use shorter test time and pre-selection was tested. Test length was varied and compared with average termination at 100 kg, termination of the test at 90 kg gives, e.g. 2% reduction in accuracy of estimated breeding values (EBV) for both breeds and termination at 80 kg gives 5% reduction in accuracy of EBVs for Landrace and 3% for Duroc. A shorter test period can decrease test costs per boar, but also gives possibilities to increase selection intensity as there will be room for testing more boars.


Fatal necrosis of the liver during ABVD chemotherapy for Hodgkin's disease. A case report.
A case of massive fatal liver necrosis during chemotherapy for stage IVA Hodgkin's disease is described. A previously healthy 50-year-old male was given doxorubicin 25 mg/m2, bleomycin 10 mg/m2 and vinblastine 6 mg/m2 on days 1 and 14 of the cycle combined with dacarbazine 150 mg/m2 on days 1-5 (ABVD) with 4-week intervals. During the fourth cycle of chemotherapy the patient developed fatal liver necrosis with anuria and uncontrolled bleeding. At autopsy, 80% of the liver was necrotic and viable cells were seen only in periportal areas. Dacarbazine may have caused the necrosis. Liver necrosis caused by drugs should be considered if unexplained hepatomegaly and a rise in serum liver enzyme levels is noted during ABVD treatment.


A new control method of nonlinear systems based on impulse responses of universal learning networks.
A new control method of nonlinear dynamic systems is proposed based on the impulse responses of universal learning networks (ULNs), ULNs form a superset of neural networks. They consist of a number of interconnected nodes where the nodes may have any continuously differentiable nonlinear functions in them and each pair of nodes can be connected by multiple branches with arbitrary time delays. A generalized learning algorithm is derived for the ULNs, in which both the first order derivatives (gradients) and the higher order derivatives are incorporated. One of the distinguished features of the proposed control method is that the impulse response of the systems is considered as an extended part of the criterion function and it can be calculated by using the higher order derivatives of ULNs. By using the impulse response as the criterion function, nonlinear dynamics with not only quick response but also quick damping and small steady state error can be more easily obtained than the conventional nonlinear control systems with quadratic form criterion functions of state and control variables.


Hydrogen-rich saline attenuates isoflurane-induced caspase-3 activation and cognitive impairment via inhibition of isoflurane-induced oxidative stress, mitochondrial dysfunction, and reduction in ATP levels.
The inhaled general anesthetic isoflurane has been shown to induce caspase-3 activation in vitro and in vivo. The underlying mechanisms and functional consequences of this activity remain unclear. Isoflurane can induce caspase-3 activation by causing accumulation of reactive oxygen species (ROS), mitochondrial dysfunction, and reduction in adenosine triphosphate (ATP) levels. This study aimed to investigate the protective effect of hydrogen, a novel antioxidant, against isoflurane-induced caspase-3 activation and cognitive impairment. H4 human neuroglioma cells overexpressing human amyloid precursor protein were treated with saline or hydrogen-rich saline (HS, 300 μM), with or without 2% isoflurane, for 6 h or 3 h. Western blot analysis, fluorescence assays, and a mitochondrial swelling assay were used to evaluate caspase-3 activation, levels of ROS and ATP, and mitochondrial function. The effect of the interaction of isoflurane (1.4% for 2 h) and HS (5 mL/kg) on cognitive function in mice was also evaluated using a fear conditioning test. We found that HS attenuated isoflurane-induced caspase-3 activation. Moreover, HS treatment mitigated isoflurane-induced ROS accumulation, opening of mitochondrial permeability transition pores, reduction in mitochondrial membrane potential, and reduction in cellular ATP levels. Finally, HS significantly alleviated isoflurane-induced cognitive impairment in mice. Our results suggest that HS attenuates isoflurane-induced caspase-3 activation and cognitive impairment via inhibition of isoflurane-induced oxidative stress, mitochondrial dysfunction, and reduction in ATP levels. These findings warrant further research into the underlying mechanisms of this activity, and indicate that HS has the potential to attenuate anesthesia neurotoxicity.


Computational Design of Multinuclear Metalloproteins Using Unnatural Amino Acids.
Multinuclear metal ion clusters, coordinated by proteins, catalyze various critical biological redox reactions, including water oxidation in photosynthesis, and nitrogen fixation. Designed metalloproteins featuring synthetic metal clusters would aid in the design of bio-inspired catalysts for various applications in synthetic biology. The design of metal ion-binding sites in a protein chain requires geometrically constrained and accurate placement of several (between three and six) polar and/or charged amino acid side chains for every metal ion, making the design problem very challenging to address. Here, we describe a general computational method to redesign oligomeric interfaces of symmetric proteins for the purpose of creating novel multinuclear metalloproteins with tunable geometries, electrochemical environments, and metal cofactor stability via first and second-shell interactions. The method requires a target symmetric organometallic cofactor whose coordinating ligands resemble the side chains of a natural or unnatural amino acid and a library of oligomeric protein structures featuring the same symmetry as the target cofactor. Geometric interface matches between target cofactor and scaffold are determined using a program that we call symmetric protein recursive ion-cofactor sampler (SyPRIS). First, the amino acid-bound organometallic cofactor model is built and symmetrically aligned to the axes of symmetry of each scaffold. Depending on the symmetry, rigid body and inverse rotameric degrees of freedom of the cofactor model are then simultaneously sampled to locate scaffold backbone constellations that are geometrically poised to incorporate the cofactor. Optionally, backbone remodeling of loops can be performed if no perfect matches are identified. Finally, the identities of spatially proximal neighbor residues of the cofactor are optimized using Rosetta Design. Selected designs can then be produced in the laboratory using genetically incorporated unnatural amino acid technology and tested experimentally for structure and catalytic activity.


An investigation of emotion recognition training to reduce symptoms of social anxiety in adolescence.
This study aimed to examine the effect of emotion recognition training on social anxiety symptoms among adolescents, aged 15-18 years. The study included a screening session, which identified participants who scored above a cut-off on a self-report measure of social anxiety for enrolment into a randomized controlled trial (Clinical Trials ID: NCT02550379). Participants were randomized to an intervention condition designed to increase the perception of happiness over disgust in ambiguous facial expressions or a sham intervention control condition, and completed self-report measures of social anxiety, fear of negative evaluation, anxiety-related disorders, and depressive symptoms. The intervention group demonstrated a strong shift in the balance point at which they perceived happiness over disgust in ambiguous facial expressions. This increase in positive perception was not associated with any changes in the primary outcome of social anxiety; however, some evidence of improvement in symptomatology was observed on one of a number of secondary outcomes. Those in the intervention group had lower depression symptoms at 2-week follow-up, compared to those in the control group who received the sham intervention training. Potential reasons for why the shift in balance point measurement was not associated with a concurrent shift in symptoms of social anxiety are discussed.


Binding characteristics of brain-derived neurotrophic factor to its receptors on neurons from the chick embryo.
Brain-derived neurotrophic factor (BDNF), a protein known to support the survival of embryonic sensory neurons and retinal ganglion cells, was derivatized with 125I-Bolton-Hunter reagent and obtained in a biologically active, radioactive form (125I-BDNF). Using dorsal root ganglion neurons from chick embryos at 9 d of development, the basic physicochemical parameters of the binding of 125I-BDNF with its receptors were established. Two different classes of receptors were found, with dissociation constants of 1.7 x 10(-11) M (high-affinity receptors) and 1.3 x 10(-9) M (low-affinity receptors). Unlabeled BDNF competed with 125I-BDNF for binding to the high-affinity receptors with an inhibition constant essentially identical to the dissociation constant of the labeled protein: 1.2 x 10(-11) M. The association and dissociation rates from both types of receptors were also determined, and the dissociation constants calculated from these kinetic experiments were found to correspond to the results obtained from steady-state binding. The number of high-affinity receptors (a few hundred per cell soma) was 15 times lower than that of low-affinity receptors. No high-affinity receptors were found on sympathetic neurons, known not to respond to BDNF, although specific binding of 125I-BDNF to these cells was detected at a high concentration of the radioligand. These results are discussed and compared with those obtained with nerve growth factor on the same neuronal populations.


Flexibility in the use of requesting gestures in squirrel monkeys (Saimiri sciureus).
Three squirrel monkeys, trained to make a requesting gesture, were tested in the presence of a human assistant whose visual attention varied across trials. When food was available in one dish and an empty dish was nearby, the monkeys pointed overwhelmingly toward the former, regardless of where the assistant was looking. Looking at the assistant while pointing ("monitoring") peaked when she looked at them and when she attempted to engage them in joint attention. When only one dish was present, the monkeys refrained from gesturing if it was empty and if no assistant was present. They gestured more when the assistant made eye contact with them. Furthermore, when the assistant's focus of attention switched from the dish or the ceiling to the monkeys, the latter resumed pointing and increased their monitoring of the assistant. This is the first demonstration of such flexible use of an intentionally communicative requesting gesture in New World monkeys.


Longitudinal development of the human white matter structural connectome and its association with brain transcriptomic and cellular architecture.
From childhood to adolescence, the spatiotemporal development pattern of the human brain white matter connectome and its underlying transcriptomic and cellular mechanisms remain largely unknown. With a longitudinal diffusion MRI cohort of 604 participants, we map the developmental trajectory of the white matter connectome from global to regional levels and identify that most brain network properties followed a linear developmental trajectory. Importantly, connectome-transcriptomic analysis reveals that the spatial development pattern of white matter connectome is potentially regulated by the transcriptomic architecture, with positively correlated genes involve in ion transport- and development-related pathways expressed in excitatory and inhibitory neurons, and negatively correlated genes enriches in synapse- and development-related pathways expressed in astrocytes, inhibitory neurons and microglia. Additionally, the macroscale developmental pattern is also associated with myelin content and thicknesses of specific laminas. These findings offer insights into the underlying genetics and neural mechanisms of macroscale white matter connectome development from childhood to adolescence.


Megavoltage CT on a tomotherapy system.
A megavoltage computed tomography (MVCT) system was developed on the University of Wisconsin tomotherapy benchtop. This system can operate either axially or helically, and collect transmission data without any bounds on delivered dose. Scan times as low as 12 s per slice are possible, and scans were run with linac output rates of 100 MU min(-1), although the system can be tuned to deliver arbitrarily low dose rates. Images were reconstructed with clinically reasonable doses ranging from 8 to 12 cGy. These images delineate contrasts below 2% and resolutions of 3.0 mm. Thus, the MVCT image quality of this system should be sufficient for verifying the patient's position and anatomy prior to radiotherapy. Additionally, synthetic data were used to test the potential for improved MVCT contrast using maximum-likelihood (ML) reconstruction. Specifically, the maximum-likelihood expectation-maximization (ML-EM) algorithm and a transmission ML algorithm were compared with filtered backprojection (FBP). It was found that for expected clinical MVCT doses enough imaging photons are used such that little benefit is conferred by the improved noise model of ML algorithms. For significantly lower doses, some quantitative improvement is achieved through ML reconstruction. Nonetheless, the image quality at those lower doses is not satisfactory for radiotherapy verification.


Supercritical fluids processing of recombinant human growth hormone.
The aim of the study was to investigate the feasibility of precipitating recombinant human growth hormone (hGH) from aqueous solutions using conventional and modified techniques of solution-enhanced dispersion (SEDS) by supercritical fluids. The study investigated the effect on hGH stability of adding isopropanol either as a cosolvent with the original aqueous protein solution (conventional process) or to the supercritical carbon dioxide before mixing with the aqueous protein solution (modified process). The influence of the addition of sucrose (with or without isopropanol) on the precipitation behavior and stability of the protein was also studied. Experiments were performed under various processing conditions (pressure 100-200 bars and temperature 40-50 degrees C), and with various flow rates and solution compositions (CO2/isopropanol and protein solution). Bioanalytical characterization of the resulting powders involved spectrophotometry, sodium dodecyl sulfate-polycrylamide gel electrophoresis, reverse-phase high performance liquid chromatography (RP-HPLC), and size exclusion chromatography. Solid-state characterization was performed using differential scanning calorimetry, X-ray powder diffraction, scanning electron microscopy, and Karl Fischer techniques. Results showed that with both conventional and modified methods, under optimum processing conditions, the presence of sucrose in the solution decreased the destabilizing effects of the solvent and/or process on the structure of hGH. More hGH was dissolved from the precipitated powders containing sucrose than from those containing only isopropanol. Reverse-phase HPLC indicated that about 94% of the hGH was recovered in its native form. The proportion of dimers and oligomers was reduced in the presence of sucrose; about 92% of the soluble protein was present in monomer form under optimal conditions. The remaining undissolved protein was in monomeric form. The precipitated powders were amorphous, containing particulate aggregates in the size range 1-6 microm with 5-10% residual moisture content. In conclusion, hGH was successfully precipitated from aqueous solution using SEDS technology. The presence of sucrose in the protein solution promoted the precipitation of hGH and reduced aggregation and improved dissolution.


Proteomic analysis and antivenomics study of Western India Naja naja venom: correlation between venom composition and clinical manifestations of cobra bite in this region.
Background: Snakebite is a severe problem in the tropical countries including Indian subcontinent. Premier cases of cobra bites are being reported from western India (WI). Research design and methods: The proteome of WI N. naja venom (NnV) was deciphered by high resolution mass spectrometry analysis of venom, further fractionated by gel filtration (GF) or RP-HPLC followed by SDS-PAGE and then tandem mass spectrometric analysis of protein bands. The efficacy of commercial polyantivenom (PAV) towards WINnV was assessed by ELISA, immuno-blot, neutralization, and venom-PAV immunoaffinity chromatography studies. Results: Proteomic analysis of WINnV, GF fractions, and SDS-PAGE protein bands of RP-HPLC and GF peaks identified 14, 34, 40, and 54, distinct proteins, respectively, when searched against Elapidae database. The biochemical properties of WINnV correlated well with its proteome composition and pathophysiology of cobra envenomation, including neuroparalysis. This study also highlighted the differences in proteome composition between WINnV and previously reported Eastern India NnV. The tested antivenoms exhibited poor immuno-recognition and neutralization of low molecular mass proteins (<20 kDa), such as three-finger toxins, the major class of protein in WINnV. Conclusion: Improvements in production protocols of antivenoms is the necessity of the hour, supplemented with antibodies raised against the poorly recognized toxins.


Mechanistic Modelling of DNA Repair and Cellular Survival Following Radiation-Induced DNA Damage.
Characterising and predicting the effects of ionising radiation on cells remains challenging, with the lack of robust models of the underlying mechanism of radiation responses providing a significant limitation to the development of personalised radiotherapy. In this paper we present a mechanistic model of cellular response to radiation that incorporates the kinetics of different DNA repair processes, the spatial distribution of double strand breaks and the resulting probability and severity of misrepair. This model enables predictions to be made of a range of key biological endpoints (DNA repair kinetics, chromosome aberration and mutation formation, survival) across a range of cell types based on a set of 11 mechanistic fitting parameters that are common across all cells. Applying this model to cellular survival showed its capacity to stratify the radiosensitivity of cells based on aspects of their phenotype and experimental conditions such as cell cycle phase and plating delay (correlation between modelled and observed Mean Inactivation Doses R(2) > 0.9). By explicitly incorporating underlying mechanistic factors, this model can integrate knowledge from a wide range of biological studies to provide robust predictions and may act as a foundation for future calculations of individualised radiosensitivity.


Mitogen stimulation during rosette formation: a test to measure numbers and function of T, B, 'D' and 'null' lymphocytes in man.
A technically simple procedure is described for simultaneous determination of numbers and functions of human blood lymphocyte subpopulations. The method is based on stimulation with mitogens (PHA, Con A and PWM) with simultaneous rosetting to detect E receptors and C3 receptors. Con A and PWM activate the affinity of C3 receptors in normal lymphocytes increasing the number of cells rosetting with zymosan-complement particles, and also the number of cells giving rosettes with both zymosan-complement particles and sheep red blood cells (B cells and 'D' cells respectively). PHA activates E receptors, increasing the number of T cells detected. The lymphocytes of 33 patients with allergic diseases of the skin and respiratory tract showed a decrease in C3 rosettes during stimulation with Con A and PWM, which suggests a defect of helper and suppressor functions of lymphocytes in these patients.


Pharmacological manipulation of immune alterations induced by an aversive conditioned stimulus: evidence for a beta-adrenergic receptor-mediated Pavlovian conditioning process.
The effect of propranolol, a beta-adrenergic receptor antagonist, on the suppression of splenic mitogenic responsiveness induced by an aversive conditioned stimulus (CS) was evaluated in rats. Experiment 1 demonstrated that propranolol administration before presentation of the CS completely blocked the suppressive effect. In contrast, Experiment 2 showed that administration of propranolol during the training of the aversive CS had no effect on the suppressive effect of the CS in a subsequent test. These findings indicate that the release of catecholamines is responsible for the conditioned immune alteration of splenic lymphocytes. Taken further, these findings suggest that the learning of the conditioned immunomodulatory response to an aversive CS is a Pavlovian conditioning process that is not dependent on the performance of the conditioned response during training.


Mutations to Less-Preferred Synonymous Codons in a Highly Expressed Gene of Escherichia coli: Fitness and Epistatic Interactions.
Codon-tRNA coevolution to maximize protein production has been, until recently, the dominant hypothesis to explain codon-usage bias in highly expressed bacterial genes. Two predictions of this hypothesis are 1) selection is weak; and 2) similar silent replacements at different codons should have similar fitness consequence. We used an allele-replacement strategy to change five specific 3rd-codon-position (silent) sites in the highly expressed Escherichia coli ribosomal protein gene rplQ from the wild type to a less-preferred alternative. We introduced the five mutations within a 10-codon region. Four of the silent sites were chosen to test the second prediction, with a CTG to CTA mutation being introduced at two closely linked leucine codons and an AAA to AAG mutation being introduced at two closely linked lysine codons. We also introduced a fifth silent mutation, a GTG to GTA mutation at a valine codon in the same genic region. We measured the fitness effect of the individual mutations by competing each single-mutant strain against the parental wild-type strain, using a disrupted form of the araA gene as a selectively neutral phenotypic marker to distinguish between strains in direct competition experiments. Three of the silent mutations had a fitness effect of |s| > 0.02, which is contradictory to the prediction that selection will be weak. The two leucine mutations had significantly different fitness effects, as did the two lysine mutations, contradictory to the prediction that similar mutations at different codons should have similar fitness effects. We also constructed a strain carrying all five silent mutations in combination. Its fitness effect was greater than that predicted from the individual fitness values, suggesting that negative synergistic epistasis acts on the combination allele.


Novel hypomyelinating leukoencephalopathy affecting early myelinating structures: clinical course in two brothers.
A novel leukoencephalopathy, termed hypomyelinating leukoencephalopathy affecting early myelinating structures (HEMS), was recently described. Here we report on two patients affected by HEMS with a clinical picture characterized by early-onset nystagmus and thereafter progressive cerebellar signs and mild spasticity predominantly affecting the legs. In one patient, who has the longest follow-up described to date, we detected a mild worsening of the clinical and neuroradiologic picture after a long period of stability lasting until age 6 years. The most recent magnetic resonance image, performed at the age of 11 years, showed a more severe neuroradiologic picture characterized by involvement of almost the entire supratentorial white matter, with relative sparing of the subcortical fibers. We also provide spectroscopy results, not previously reported in this disorder, that support the idea of a progressive disease course on neuroimaging. Our findings suggest that HEMS patients should undergo a new magnetic resonance imaging evaluation after a certain interval to look for possible progression of the abnormalities.


[Determination of cartilage thickness in the ankle joint. an MRT (1.5)-anatomical comparative study].
The aim of this study was to evaluate the accuracy of MRI in the measurement of cartilage thickness of the ankle joint in comparison to pathologic and histologic specimens. The ankle joints of four fresh cadaver feet were imaged on a 1.5T MR-unit in the coronal plane. Standard T1-weighted spin echo (SE) and a T1-weighted 3D-GE (FLASH-3D) sequence with fat saturation were applied. Following MR imaging, the talus was explanted and cut parallel to the MR images for macroscopic evaluation and histologic specimens were prepared. MRI measurements of the cartilage thickness of the talus were carried out in two ways: without and with consideration of a transition zone of intermediate signal intensity between hyperintense cartilage and hypointense cortical bone. The data were compared with the anatomic specimens as the gold standard expressing the difference as a percentage value. On histologic specimens thickness of deep calcified cartilage layer was measured. Measurements without the transition zone showed a mean underestimation of talus cartilage thickness of 46.8% (41.3-52.1) for T1-SE and 47.5% (43.1-52.1) for fat saturated FLASH-3D images. Considering the transition zone the mean values were 25.0% (23.1-26.2) and 14.1% (6.7-21.5). The histologic specimens showed a three-fold increase of thickness of deep calcified cartilage layer. Measurements of the cartilage layer of the ankle joint on MR images are only accurate if the transition zone (calcified cartilage layer) is considered and the optimal pulse sequence (FLASH-3D-fat-sat) is applied.


Myosin functional domains encoded by alternative exons are expressed in specific thoracic muscles of Drosophila.
The Drosophila 36B muscle myosin heavy chain (MHC) gene has five sets of alternatively spliced exons that encode functionally important domains of the MHC protein and provide a combinatorial potential for expression of as many as 480 MHC isoforms. In this study, in situ hybridization analysis has been used to examine the complexity and muscle specificity of MHC isoform expression in the fibrillar indirect flight muscle (IFM), the tubular direct flight muscles (DFM) and tubular tergal depressor of the trochanter muscle (TDT), and the visceral esophageal muscle in the adult thorax. Our results show that alternative splicing of the MHC gene transcripts is precisely regulated in these thoracic muscles, which express three MHC isoforms. Individual thoracic muscles each express transcripts of only one isoform, as detectable by in situ hybridization. An apparently novel fourth MHC isoform, with sequence homology to the rod but not to the head domain of the 36B MHC, is expressed in two direct flight muscles. These findings form a basis for transgenic experiments designed to analyze the muscle-specific functions of MHC domains encoded by alternative exons.


ABO Blood Groups Are Not Associated With COVID-19 Disease Incidence and Severity When Correcting for Ethnicity Differences in Blood Type.
To determine if blood type is a risk factor for coronavirus disease 2019 (COVID-19) disease incidence and severity after correcting for ethnicity differences between novel infections and known ABO blood type frequency differences. We performed a retrospective analysis on all severe acute respiratory system coronavirus 2 (SARS-CoV-2) infections and disease severity across two major testing sites in Colorado. We evaluated all individuals with a SARS-CoV-2 nucleic acid test (NAT) and a known blood type between March 1, 2020, and June 1, 2020. We then created a prediction algorithm based on the corrected blood types by ethnicity using data from the Colorado Department of Health and established blood types by ethnicity. We applied this prediction algorithm to all patients in our sample. Of 8,676 patients, 485 (5.6%) had a positive SARS-CoV-2 NAT test and 8,191 (94.4%) had a negative test. All patients had ABO blood types that mirrored the expected blood type distribution within the state of Colorado (P = .15, χ 2 statistic = 5.31). No differences in expected blood groups were present between ethnicity-adjusted SARS-CoV-2-negative and SARS-CoV-2-positive patients (χ 2 = 3.416313, P = .332). Blood type is not associated with COVID-19 disease incidence or severity after correcting for ethnicity differences in expected blood type frequencies.


Deep learning-enabled breast cancer endocrine response determination from H&E staining based on ESR1 signaling activity.
Estrogen receptor (ER) positivity by immunohistochemistry has long been a main selection criterium for breast cancer patients to be treated with endocrine therapy. However, ER positivity might not directly correlate with activated ER signaling activity, which is a better predictor for endocrine therapy responsiveness. In this study, we investigated if a deep learning method using whole-slide H&E-stained images could predict ER signaling activity. First, ER signaling activity score was determined using RNAseq data available from each of the 1082 breast cancer samples in the TCGA Pan-Cancer dataset based on the Hallmark Estrogen Response Early gene set from the Molecular Signature Database (MSigDB). Then the processed H&E-stained images and ER signaling activity scores from a training cohort were fed into ResNet101 with three additional fully connected layers to generate a predicted ER activity score. The trained models were subsequently applied to an independent testing cohort. The result demonstrated that ER + /HER2- breast cancer patients with a higher predicted ER activity score had longer progression-free survival (p = 0.0368) than those with lower predicted ER activity score. In conclusion, a convolutional deep neural network can predict prognosis and endocrine therapy response in breast cancer patients based on whole-slide H&E-stained images. The trained models were found to robustly predict the prognosis of ER + /HER2- patients. This information is valuable for patient management, as it does not require RNA-seq or microarray data analyses. Thus, these models can reduce the cost of the diagnosis workflow if such information is required.


Chemoport-A Savior in Children Who Require Chronic Venous Access: An Observational Study.
Long-term venous access is cumbersome in children because of their thin caliber veins, less cooperative nature, and easy compromise of venous integrity. Hence, a study was conducted to evaluate the indication, efficacy, and safety of chemoport in children who require chronic venous access. Children who underwent chemoport insertion between January 2008 and December 2017 were retrospectively evaluated. A total of 159 children (169 chemoports) were included in the study. The most common indication for chemoport insertion was acute lymphoblastic leukemia (51.5%). The mean chemoport days were 832±666 days. Among the 169 chemoports, 55.0% were removed after treatment completion. The chemoport was not removed in 35.5% of the patients, as 28.4% of the patients were still under treatment and 7.1% died during the treatment. Sixteen patients (0.1 per 1,000 chemoport days) had a premature chemoport removal. The indications were port-related bloodstream infection (12 patients), port pocket infection (1 patient), exposed chemoport (1 patient), and blocked chemoport catheter (2 patients). Twenty-two patients (0.15 per 1,000 chemoport days) had complications of port-related bloodstream infection (0.09 per 1,000 chemoport days), making it the most common. Other complications include block, fracture, arrhythmias, avulsion, bleeding, decubitus-over-port, and port pocket infection. Owing to the safe, reliable, and low complication rate of chemoports, more children can be saved from deadly illnesses. Chemoport is the best option for children who require chronic venous access.


Surgery after intracranial investigation with subdural electrodes in patients with drug-resistant focal epilepsy: outcome and complications.
Video-EEG monitoring with intracranial subdural electrodes is a useful assessment tool for the localization of the epileptogenic zone in patients with drug-resistant focal epilepsy. We aimed at assessing the morbidity related to electrode implantation and the surgical outcome in patients who underwent epilepsy surgery after intracranial EEG monitoring. All patients (N = 58) admitted to our Epilepsy Surgery Centre for drug-resistant focal epilepsy who underwent resective surgery after intracranial monitoring with subdural electrodes and were followed up for at least 2 years were included in the study. Their mean age was 30.4 years (range 8-60 years), 25 (43 %) were female, and 44 (76 %) had a preoperatively detected structural lesion. The mean duration of invasive recording was 2.3 days (range 1-14 days). Extraoperative ECoG allowed the identification of the epileptogenic focus in all cases. The temporal lobe was involved in 21 (36 %) patients, whereas extratemporal foci were identified in 24 (41 %) patients. Thirteen patients (23 %) had multilobar involvement. Functional brain mapping was performed in 15 (26 %) patients. Transient complications related to electrode implantation occurred in three patients. Among patients with evidence of lesion on preoperative MRI, lesionectomy alone was performed in 12 cases (27 %), while it was combined with tailored cortical resection in the remaining cases. Tailored cortical resection was also performed in patients without evidence of lesion on MRI. After resective surgery, transient neurological deficits occurred in five cases, while another patient experienced permanent lateral homonymous hemianopia. At the last follow-up observation, 34 (57 %) patients were seizure-free (Engel class I). This study suggests that invasive EEG recording with subdural electrodes may be useful and fairly safe for many candidates for epilepsy surgery.


Perinatal antibiotic exposure of neonates in Canada and associated risk factors: a population-based study.
To describe neonatal antibiotic exposures occurring immediately before and after birth and their associated risk factors. A retrospective review of the hospital charts of 449 mother-neonate pairs enrolled in the Canadian Healthy Infant Longitudinal Development national birth-cohort study was conducted at two tertiary hospitals and one rural hospital in Manitoba, Canada. The main outcome measures included the following: maternal and neonatal antibiotic use during the perinatal period; indications for antibiotic use, including suspected neonatal sepsis, maternal group B Streptococcus (GBS), premature rupture of membranes and caesarean-section; maternal health status, focusing on gestational hypertension, gestational diabetes, obesity and primigravida pregnancies. During the perinatal period, 45.0% of neonates were exposed to antibiotics. Intravenous penicillin G (17%) and cefazolin (16%) were the most commonly administered intrapartum antibiotics. Colonization with GBS was confirmed in 21.2% of women and treated with antibiotics in 86% of cases. Overweight women and women with hypertension were significantly more likely to receive intrapartum antibiotics for caesarean section or GBS prophylaxis. Antibiotic treatment of the neonate was highest following emergency caesarean section (12%) or unknown maternal GBS status (20%). Neonates in Canada are routinely exposed to antibiotics during the perinatal period.


The Impact of Early Classroom Inattention on Phonological Processing and Word-Reading Development.
The present study investigated the longitudinal relationships between inattention, phonological processing and word reading across the first 2 years of formal reading instruction. In all, 136 school entrants were administered measures of letter knowledge, phonological awareness, phonological memory, rapid naming, and word reading at the start and end of their 1st year of school, and the end of their 2nd year, while teachers completed rating scales of inattention. School entry inattentiveness predicted unique variance in word reading at the end of first grade, after controlling for verbal ability, letter knowledge, and phonological processing. End-of-first-grade inattention predicted a small but significant amount of unique variance in second-grade word reading and word-reading efficiency. Inattention, however, was not a reliable predictor of phonological processing in either first or second grade. Early classroom inattentiveness influences learning to read independent of critical developmental precursors of word-reading development.


Using Length of Stay to Understand Patient Flow for Pediatric Inpatients.
Develop and test a new metric to assess meaningful variability in inpatient flow. Using the pediatric administrative dataset, Pediatric Health Information System, that quantifies the length of stay (LOS) in hours, all inpatient and observation encounters with 21 common diagnoses were included from the calendar year 2013 in 38 pediatric hospitals. Two mutually exclusive composite groups based on diagnosis and presence or absence of an ICU hospitalization termed Acute Care Composite (ACC) and ICU Composite (ICUC), respectively, were created. These composites consisted of an observed-to-expected (O/E) LOS as well as an excess LOS percentage (ie, the percent of day beyond expected). Seven-day all-cause risk-adjusted rehospitalizations was used as a balancing measure. The combination of the ACC, the ICUC, and the rehospitalization measures forms this new metric. The diagnosis groups in the ACC and the ICUC included 113,768 and 38,400 hospitalizations, respectively. The ACC had a median O/E LOS of 1.0, a median excess LOS percentage of 23.9% and a rehospitalization rate of 1.7%. The ICUC had a median O/E LOS of 1.1, a median excess LOS percentage of 32.3%, and rehospitalization rate of 4.9%. There was no relationship of O/E LOS and rehospitalization for either ACC or ICUC. This metric shows variation among hospitals and could allow a pediatric hospital to assess the performance of inpatient flow.


Increased lncRNA AFAP1-AS1 expression predicts poor prognosis and promotes malignant phenotypes in gastric cancer.
The clinical significance and biological functions of long non-coding RNA AFAP1-AS1 in gastric cancer (GC) remain larger elucidated. The aim of the study is to investigate the role of lncRNA AFAP1-AS1 involved in GC progression. Quantitative reverse transcription-polymerase chain reaction (QRT-PCR) assay was used to evaluate the expression of lncRNA AFAP1-AS1 in GC tissues, when compared with adjacent noncancerous tissues, respectively. Associations between lncRNA AFAP1-AS1 and the clinicopathological factors in GC patients were analyzed by X2-test. The prognostic significance was evaluated using Kaplan-Meier curve and Cox regression analysis. CCK8, flow cytometry analysis and transwell invasion assays were performed to assess cell proliferation and invasion abilities of GC. In this study, we verified that lncRNA AFAP1-AS1 expression was dramatically increased in GC tissues and cells, compared with noncancerous gastric tissues and control cells. The higher lncRNA AFAP1-AS1 expression was positively correlated with lymph node metastasis (p = 0.001), TNM stage (p = 0.006) and worse overall survival (OS) time in GC patients. Multivariate Cox analysis suggested that lymph node metastasis (Hazard ratio, HR = 2.966, p = 0.001), TNM stage (HR = 2.855, p = 0.001), and lncRNA AFAP1-AS1 expression levels (HR = 3.315, p = 0.001) were independent prognostic factors of OS in GC patients. Knockdown of lncRNA AFAP1-AS1 significantly inhibited the cell proliferation and cell cycle progression. Moreover, reduced lncRNA AFAP1-AS1 also inhibited the cell invasion ability via regulating cell epithelial-mesenchymal transition (EMT) phenomenon in GC. These results demonstrated that lncRNA AFAP1-AS1 may be a potential therapeutic target for GC.


Increased neurotransmitter release during long-term potentiation at mossy fibre-granule cell synapses in rat cerebellum.
During long-term potentiation (LTP) at mossy fibre-granule cell synapses in rat cerebellum synaptic transmission and granule cell intrinsic excitability are enhanced. Although it is clear that changes in granule cell excitability are mediated postsynaptically, there is as yet no direct evidence for the site and mechanism of changes in transmission. To approach this problem, evoked postsynaptic currents (EPSCs) and miniature synaptic currents (mEPSCs) were recorded by patch-clamp in cerebellar slices obtained from P17-P23 rats. LTP was induced by theta-burst stimulation paired with depolarization. During LTP, the EPSCs showed a significant decrease in the coefficient of variation (CV; 28.9 +/- 5.2%, n= 8; P < 0.002), the number of failures (87.1 +/- 41.9%, n= 8; P < 0.04), and the paired-pulse ratio (PPR; 25.5 +/- 4.1% n= 5; P < 0.02). Similar changes were observed by increasing neurotransmitter release (extracellular solutions with high Ca(2+)/Mg(2+) ratio), whereas increases in CV, numbers of failures and PPR occurred when release was decreased (extracellular solutions with low Ca(2+)/Mg(2+) ratio; 10 microm Cl-adenosine). No changes followed modifications of postsynaptic holding potentials, while CV and failures were reduced when the number of active synapses was increased. LTP was prevented by use of solutions with high Ca(2+)/Mg(2+) ratio. Moreover, LTP and the associated CV decrease were observed in the spillover-mediated component of AMPA EPSCs and in NMDA EPSCs. During LTP, mEPSCs did not change in amplitude or variability but significantly increased in frequency (47.6 +/- 16%, n= 4; P < 0.03). By binomial analysis changes in EPSCs were shown to be due to increased release probability (from 0.6 +/- 0.08 to 0.73 +/- 0.06, n= 7; P < 0.02) with a constant number of three to four releasing sites. These observations provide evidence for increased neurotransmitter release during LTP at mossy fibre-granule cell synapses.


International issues in the supply of tobacco: recent changes and implications for alcohol.
This paper reviews international issues in the supply of tobacco and tobacco products, including trade liberalization and globalization. The paper begins with a brief discussion of the theoretical foundations for trade and trade restrictions. This is followed by a description of the treatment of tobacco and tobacco products in recent multi-lateral, regional and bilateral trade agreements, as well as a short discussion of the recent globalization of the tobacco industry. Included in this description is a review of the empirical evidence on the impact of trade liberalization on tobacco use. The implications of two recently proposed international agreements--the Multilateral Agreement on Investments and the Framework Convention for Tobacco Control--are then discussed. The paper concludes by summarizing the theoretical and empirical evidence that shows clearly that trade liberalization has significantly increased tobacco use, particularly in low and middle-income countries, and follows this with a discussion of the lessons learned from tobacco for controlling alcohol supply.


Platelet sequestration during hypothermia in dogs treated with sulphinpyrazone and ticlopidine--reversibility accelerated after intra-abdominal rewarming.
During body cooling, a progressive thrombocytopenia is observed, with platelet sequestration mainly in the liver. Platelets return progressively to the circulation during rewarming. In this work the presence of platelet clumps is demonstrated by electron microscopy inside the hepatic sinusoids of dogs cooled to 20 degrees C by immersion in iced water. Such clumps were not found either before cooling or after body rewarming. Similar platelet clumps in the hepatic microcirculation were found in two other groups of dogs cooled and rewarmed as before, but previously treated with the antiaggregants sulphinpyrazone or ticlopidine. Another group of dogs was cooled and rewarmed by intra-abdominal circulation of physiological saline respectively at 4 degrees C and 40 degrees C. In these animals a similar decrease of circulating platelets was observed. However, just after rewarming was started, with the body temperature still at 22 degrees C, sequestered platelets came back abruptly to circulation. We conclude that hepatic platelet sequestration induced by hypothermia appears mainly due to local haemodynamic conditions.


cag, a pathogenicity island of Helicobacter pylori, encodes type I-specific and disease-associated virulence factors.
cagA, a gene that codes for an immunodominant antigen, is present only in Helicobacter pylori strains that are associated with severe forms of gastroduodenal disease (type I strains). We found that the genetic locus that contains cagA (cag) is part of a 40-kb DNA insertion that likely was acquired horizontally and integrated into the chromosomal glutamate racemase gene. This pathogenicity island is flanked by direct repeats of 31 bp. In some strains, cag is split into a right segment (cagI) and a left segment (cagII) by a novel insertion sequence (IS605). In a minority of H. pylori strains, cagI and cagII are separated by an intervening chromosomal sequence. Nucleotide sequencing of the 23,508 base pairs that form the cagI region and the extreme 3' end of the cagII region reveals the presence of 19 ORFs that code for proteins predicted to be mostly membrane associated with one gene (cagE), which is similar to the toxin-secretion gene of Bordetella pertussis, ptlC, and the transport systems required for plasmid transfer, including the virB4 gene of Agrobacterium tumefaciens. Transposon inactivation of several of the cagI genes abolishes induction of IL-8 expression in gastric epithelial cell lines. Thus, we believe the cag region may encode a novel H. pylori secretion system for the export of virulence determinants.


Using atypical symptoms and red flags to identify non-demyelinating disease.
Red flags and atypical symptoms have been described as being useful in suggesting alternative diagnoses to multiple sclerosis (MS) and clinically isolated syndrome (CIS); however, their diagnostic utility has not been assessed. The aim of this study was to establish the predictive value of red flags and the typicality/atypicality of symptoms at presentation in relation to the final diagnosis of patients referred with suspected MS. All patients referred with suspected MS over a 3-year period were assessed by the typicality of the clinical presentation and the occurrence of red flags in relation to the eventual diagnosis. The extent of agreement of trainee and consultant neurologists as to typicality of clinical presentations was determined. Of 244 patients referred, 119 (49%) had MS/CIS and 125 (51%) did not. 41 patients were referred because of an abnormal MRI. Of 203 with clinical symptoms, 96 patients had atypical symptoms of whom, 81 (84%) did not have MS and 15 (16%) had MS/CIS. Typical symptoms occurred in 107 patients; 10% did not have MS/CIS. Atypical symptoms had a sensitivity of 84%, specificity of 90% and positive likelihood ratio (PLR) of 8.4, whereas red flags had a sensitivity of 47%, specificity of 88% and PLR of 3.9 for the exclusion of MS/CIS. Mean percentage agreement between consultants and trainees was 73% with a range of 32-96%. Atypical features at presentation are more sensitive, specific and have a higher PLR than red flags to refute a diagnosis of MS/CIS.


Engineering self-assembled 2D nano-network membranes through hierarchical phase separation for efficient air filtration.
Air pollution has garnered significant worldwide attention; however, the existing air filtration materials still suffer from issues related to monotonous structure and the inherent trade-off between PM rejection and air permeability. Herein, a spider web-inspired composite membrane with continuous monolayer structured 2D nano-networks tightly welded on nanofibers in the electrospun membrane scaffold is designed via a hierarchical phase separation strategy. The resultant biomimetic hierarchical-structured membranes possess the integrated features of hierarchical multiscale structures of 2D ultrafine networks composed of nanowires with a diameter of 31 nm self-assembled by nanoparticles, exceptional characteristics involving small average aperture, extremely low network thickness, high porosity and promising pore channel connectivity, combined with rich surface polar functional groups (3.02D dipole moment). Consequently, the composite membrane exhibits a high PM0.3 capture efficiency of 99.6 % and low pressure drop of 58.8 Pa, less than 0.06 % of atmosphere pressure, with outstanding long-term PM2.5 recycling filtration performance. The hierarchical phase separation-driven 2D nano-networks construction strategy, by virtue of their feasibility and tunability, holds great promise for widespread application across diverse membrane-related domains for air filtration.


Dellen associated with filtering blebs.
Dellen may occur in the corneas of eyes with filtering blebs. Nine cases of such dellen were encountered in a series of 97 consecutive trabeculectomies. In two such patients, the dell progressed to a deep corneal ulcer, although in most cases it healed uneventfully with minor changes in therapy. The use of topical prednisolone acetate during the postoperative period may also contribute to dellen formation by inhibiting corneal epithelial wound healing. However, the most important factor predisposing to dellen formation was poor tear-film integrity adjacent to the bleb. The secretion of the aqueous tears seemed to have little or no effect on dellen formation. Almost all dellen occurred in eyes with large filtering blebs.


Color Orchestra: Ordering Color Palettes for Interpolation and Prediction.
Color theme or color palette can deeply influence the quality and the feeling of a photograph or a graphical design. Although color palettes may come from different sources such as online crowd-sourcing, photographs and graphical designs, in this paper, we consider color palettes extracted from fine art collections, which we believe to be an abundant source of stylistic and unique color themes. We aim to capture color styles embedded in these collections by means of statistical models and to build practical applications upon these models. As artists often use their personal color themes in their paintings, making these palettes appear frequently in the dataset, we employed density estimation to capture the characteristics of palette data. Via density estimation, we carried out various predictions and interpolations on palettes, which led to promising applications such as photo-style exploration, real-time color suggestion, and enriched photo recolorization. It was, however, challenging to apply density estimation to palette data as palettes often come as unordered sets of colors, which make it difficult to use conventional metrics on them. To this end, we developed a divide-and-conquer sorting algorithm to rearrange the colors in the palettes in a coherent order, which allows meaningful interpolation between color palettes. To confirm the performance of our model, we also conducted quantitative experiments on datasets of digitized paintings collected from the Internet and received favorable results.


Vitellogenesis in the stick insect Carausius morosus I. Specific protein synthesis during ovarian development.
Vitellogenesis in the stick insect Carausius morosus (Br.) has been studied with the goal of identifying vitellogenin in various tissues. Following exposure to in vivo to radioactive amino acids, oocytes in the medium size range are labelled with a minimum delay of 6 h after the time of injection. Incorporation of radioactivity under these conditions is shown to depend upon accumulation of proteins rather than on a differential rate of protein synthesis in succeeding stages of oogenesis. By immunochemical analyses, it is shown that at least two antigens are common to both haemolymph and ovary and that one of these is also present in the fat body. Both antigens are labelled during exposure to radioactive amino acids. When analysed by the SDS polyacrylamide gel electrophoresis, extracts from both haemolymph and ovary appear to share a number of protein fractions which range in molecular weight from 40 000 to 200 000 Daltons. The labelling pattern exhibited by these fractions is clearly indicative of a protein transfer from the fat body to the oocyte. Fat body cultured in vivo for up to 4 h releases a major macromolecular complex in the external medium. The latter has been identified as vitellogenin by both immuno-precipitation assay and SDS polyacrylamide gel electrophoresis. The protein which is synthesized and secreted under these conditions results from the processing of a protein complex of higher molecular weight.


(68)Ga-DOTA-NOC PET/CT in comparison with CT for the detection of bone metastasis in patients with neuroendocrine tumours.
To retrospectively evaluate the sensitivity, specificity and accuracy of (68)Ga-DOTA-NOC PET/CT and CT alone for the evaluation of bone metastasis in patients with neuroendocrine tumour (NET). From among patients with NET who underwent (68)Ga-DOTA-NOC PET/CT between April 2006 and November 2008 in our centre, 223 were included in the study. Criteria for inclusion were pathological confirmation of NET and a follow-up period of at least 10 months. PET and CT images were retrospectively reviewed by two nuclear medicine specialists and two radiologists, respectively, without knowledge of the patient history or the findings of other imaging modalities. PET data were compared with the CT findings. Interobserver agreement was evaluated in terms of the kappa score. Clinical and imaging follow-up were used as the standard of reference to evaluate the PET findings. PET was performed for staging (49/223), unknown primary tumour detection (24/223), restaging (32/223), restaging before radioimmunotherapy (1/223), evaluation during therapy (12/223), equivocal findings on conventional imaging (4/223 at the bone level; 61/223 at sites other than bone), and follow-up (40/223). A very high interobserver agreement was observed. CT detected at least one bone lesion in only 35 of 44 patients with a positive PET scan. In particular, PET showed more lesions in 20/35 patients, a lower number of lesions in 8/35, and the same number in 7/35. The characteristics of the lesions (sclerotic, lytic, mixed) on the basis of the CT report did not influence PET reading. PET revealed the presence of at least one bone metastasis in nine patients with a negative CT scan. Considering patients with a negative PET scan (179), CT showed equivocal findings at the bone level in three (single small sclerotic abnormality in two at the spine level, and bilateral small sclerotic abnormalities in the humeri, femurs and scapula). Clinical follow-up confirmed the PET findings in all patients; thus there were no false-positive or false-negative findings. Considering all patients, PET detected more lesions than CT (246 vs. 194). As compared to CT, on a patient basis PET showed a higher sensitivity (100% vs. 80%), specificity (100% vs. 98%), positive predictive value (100% vs. 92%), and negative predictive value (100% vs. 95%). In conclusion, (68)Ga DOTA-NOC PET was more accurate than CT for the identification of bone lesions and led to a change in clinical management in nine patients with a negative CT scan.


Frequency of WT1 and 11p15 constitutional aberrations and phenotypic correlation in childhood Wilms tumour patients.
In 9-17% of Wilms tumour patients a predisposing syndrome is present, in particular WT1-associated syndromes and overgrowth syndromes. Constitutional WT1 mutations or epigenetic changes on chromosome 11p15 have also been described in Wilms tumour patients without phenotypic abnormalities. Thus, the absence of phenotypic abnormalities does not exclude the presence of a genetic predisposition, suggesting that more Wilms tumour patients may have a constitutional abnormality. Therefore, we investigated the frequency of constitutional aberrations in combination with phenotype. Clinical genetic assessment, as well as molecular analysis of WT1 and locus 11p15 was offered to a single-centre cohort of 109 childhood Wilms tumour patients. Twelve patients (11%) had a WT1 aberration and eight patients (8%) had an 11p15 aberration. Of the 12 patients with a WT1 aberration, four had WAGR syndrome (Wilms tumor, aniridia, genitourinary malformations and mental retardation), one had Denys-Drash syndrome, four had genitourinary anomalies without other syndromic features and three had bilateral disease with stromal-predominant histology at young age without congenital anomalies. Of the eight patients with an 11p15 aberration, four had Beckwith-Wiedemann syndrome (BWS), two had minor features of BWS and two had no stigmata of BWS or hemihypertrophy. Constitutional WT1 or 11p15 aberrations are frequent in Wilms tumour patients and careful clinical assessment can identify the majority of these patients. Therefore, we would recommend offering clinical genetic counselling to all Wilms tumour patients, as well as molecular analysis to patients with clinical signs of a syndrome or with features that may indicate a constitutional WT1 or 11p15 aberration.


Degradation of amyloid peptide aggregates by targeted singlet oxygen delivery from a benzothiazole functionalized naphthalene endoperoxide.
Aggregate structures formed by amyloid-β (Aβ) are correlated with the progression of pathogenesis in Alzheimer's disease. Previous works have shown that photodynamic photosensitizers were effective in oxidatively degrading amyloid-β aggregates and thus decreasing their cytotoxicity under various conditions. In this work, we designed and synthesized a benzothiazole-naphthalene conjugate, with high level of structural analogy to Thioflavin T which is known to have high affinities for the amyloid peptide aggregates. The endoperoxide form (BZTN-O2) of this compound, which releases singlet oxygen with a half-life of 77 minutes at 37 °C, successfully inhibited and/or reversed amyloid aggregation. The endoperoxide is capable of singlet oxygen release without any need for light, and its charge-neutral form could allow blood-brain barrier (BBB) permeability. The therapeutic potential of such endoperoxide compounds with amyloid binding affinity is exciting.


Roles of kallikrein-2 biomarkers (free-hK2 and pro-hK2) for predicting prostate cancer progression-free survival.
Free human kallikrein 2 (free-hK2) and hK2 pro-form (pro-hK2) have been found to be increased in tumor tissues and serum from patients with prostate cancer. We established semiautomatic assays for free-hK2 and pro-hK2 using a research version of the Beckman Coulter ACCESS2 system. Serum samples from a cohort of 189 men undergoing radical prostatectomy for known high-risk disease were assayed for Prostate-Specific Antigen (PSA), free-PSA, free-hK2, and pro-hK2. Univariate Cox regression and multivariate models were used to predict both Gleason scores and progression-free survival (PFS). Free-hk2 levels ≥80 ng/L were predictive of both Gleason scores ≥7 (p = 0.04) and PFS (p = 0.03). PSA ≥8.0 µg/L also was predictive of PFS (p = 0.02). However, neither % free-PSA nor pro-hK2, when treated as continuous or cutoff variables were associated with Gleason score or PFS. Multivariable models showed that clinical stage T1c versus T2/T3, Gleason score ≥7, and PSA ≥8.0 µg/L or clinical stage T1c versus T2/T3, Gleason scores ≥7, and free-hK2 ≥80 ng/L were among the best models predicting PFS. Both free-hK2 and PSA in conjunction with clinical stage and Gleason score are good predictors of PFS in prostate cancer.


Ancient Chinese literature reveals pathways of eggplant domestication.
Changes in key traits occurring during the processes of plant domestication have long been subjects of debate. Only in the case of genetic analysis or with extensive plant remains can specific sets of changes be documented. Historical details of the plant domestication processes are rare and other evidence of morphological change can be difficult to obtain, especially for those vegetables that lack a substantial body of archaeological data. Botanical records chronicled in the ancient literature of established ancient civilizations, such as that of China, are invaluable resources for the study and understanding of the process of plant domestication. Here, the considerable body of ancient Chinese literature is used to explore the domestication process that has occurred with the eggplant (Solanum melongena), an important vegetable in Old World. Information about eggplant domestication in the ancient Chinese literature was retrieved using a variety of methods. The information obtained was then sorted by taxon, examined and taxonomic identifications verified. It was found that the earliest record of the eggplant documented in ancient Chinese literature was in a work from 59 bc. As far as is known, this is the earliest reliable and accurately dated record of eggplant in cultivation. The analysis reveals that the process of domestication of the eggplant in China involved three principal aspects of fruit quality: size, shape and taste. These traits were actively and gradually selected; fruit size changed from small to large, taste changed from not palatable to what was termed at the time sweetish, and that over time, a wider variety of fruit shapes was cultivated. The results indicate that, in addition to data gleaned from archaeology and genetics, evidence as to changes in key traits occurring during the process of plant domestication and selective forces responsible for these changes can be traced through the ancient literature in some civilizations.


Promoting de-implementation of inappropriate antimicrobial use in cardiac device procedures by expanding audit and feedback: protocol for hybrid III type effectiveness/implementation quasi-experimental study.
Despite a strong evidence base and clinical guidelines specifically recommending against prolonged post-procedural antimicrobial use, studies indicate that the practice is common following cardiac device procedures. Formative evaluations conducted by the study team suggest that inappropriate antimicrobial use may be driven by information silos that drive provider belief that antimicrobials are not harmful, in part due to lack of complete feedback about all types of clinical outcomes. De-implementation is recognized as an important area of research that can lead to reductions in unnecessary, wasteful, or harmful practices, such as excess antimicrobial use following cardiac device procedures; however, investigations into strategies that lead to successful de-implementation are limited. The overarching hypothesis to be tested in this trial is that a bundle of implementation strategies that includes audit and feedback about direct patient harms caused by inappropriate prescribing can lead to successful de-implementation of guideline-discordant care. We propose a hybrid type III effectiveness-implementation stepped-wedge intervention trial at three high-volume, high-complexity VA medical centers. The main study intervention (an informatics-based, real-time audit-and-feedback tool) was developed based on learning/unlearning theory and formative evaluations and guided by the integrated-Promoting Action on Research Implementation in Health Services (i-PARIHS) Framework. Elements of the bundled and multifaceted implementation strategy to promote appropriate prescribing will include audit-and-feedback reports that include information about antibiotic harms, stakeholder engagement, patient and provider education, identification of local champions, and blended facilitation. The primary study outcome is adoption of evidence-based practice (de-implementation of inappropriate antimicrobial use). Clinical outcomes (cardiac device infections, acute kidney injuries and Clostridioides difficile infections) are secondary. Qualitative interviews will assess relevant implementation outcomes (acceptability, adoption, fidelity, feasibility). De-implementation theory suggests that factors that may have a particularly strong influence on de-implementation include strength of the underlying evidence, the complexity of the intervention, and patient and provider anxiety and fear about changing an established practice. This study will assess whether a multifaceted intervention mapped to identified de-implementation barriers leads to measurable improvements in provision of guideline-concordant antimicrobial use. Findings will improve understanding about factors that impact successful or unsuccessful de-implementation of harmful or wasteful healthcare practices. ClinicalTrials.gov NCT05020418.


Underwater detection of tonal signals between 0.125 and 100 kHz by harbor seals (Phoca vitulina).
The underwater hearing sensitivities of two 1-year-old female harbor seals were quantified in a pool built for acoustic research, using a behavioral psychoacoustic technique. The animals were trained to respond when they detected an acoustic signal and not to respond when they did not (go/no-go response). Pure tones (0.125-0.25 kHz) and narrowband frequency modulated (tonal) signals (center frequencies 0.5-100 kHz) of 900 ms duration were tested. Thresholds at each frequency were measured using the up-down staircase method and defined as the stimulus level resulting in a 50% detection rate. The audiograms of the two seals did not differ statistically: both plots showed the typical mammalian U-shape, but with a wide and flat bottom. Maximum sensitivity (54 dB re 1 microPa, rms) occurred at 1 kHz. The frequency range of best hearing (within 10 dB of maximum sensitivity) was from 0.5 to 40 kHz (6(1/3) octaves). Higher hearing thresholds (indicating poorer sensitivity) were observed below 1 and above 40 kHz. Thresholds below 4 kHz were lower than those previously described for harbor seals, which demonstrates the importance of using quiet facilities, built specifically for acoustic research, for hearing studies in marine mammals. The results suggest that under unmasked conditions many anthropogenic noise sources and sounds from conspecifics are audible to harbor seals at greater ranges than formerly believed.


Clinical studies of food allergy in infants and children.
This was a study of 50 patients, aged 3 months to 10 years. There were 27 males and 23 females. Symptoms varied from respiratory complaints to rash, headaches, and gastrointestinal symptoms. Twenty-seven patients had runny nose, 15 wheezing, 19 frequent infections, 10 coughing, 10 ear infections, 9 rash, 6 gastrointestinal symptoms, and 6 with headaches. Laboratory evaluation showed that 16 patients had elevated IgE. Forty-six patients had IgE RAST evaluation. Of these, only 17 had positive IgE RAST for foods including corn, egg, wheat, and milk. In contrast, 32 of 46 patients had positive IgG RAST for foods including corn, egg, wheat, and milk. Elimination diet of IgE RAST-positive and IgG RAST-positive foods caused a 70% reduction of symptoms. Although the data suggest that IgG antibodies may be helpful, further studies are warranted.


Gender differences in the association between morbidity and mortality among middle-aged men and women.
We examined gender differences in mortality, morbidity, and the association between the 2. We used health data from 2 studies of middle-aged men and women: the British Whitehall II cohort of employees from 20 civil service departments in London and the 1989 French GAZEL (this acronym refers to the French gas and electric companies) of employees of France's national gas and electricity company. Participants were aged 35 to 55 years when assessed for morbidity and followed up for mortality over 17 years. Male mortality was higher than female mortality in Whitehall II (hazard ratio [HR] = 1.56; 95% confidence interval [CI] = 1.28, 1.91) and the GAZEL cohort (HR = 1.99; CI = 1.66, 2.40). Female excess morbidity was observed for some measures in the Whitehall II data and for 1 measure in the GAZEL data. Only self-reported sickness absence in the Whitehall II data was more strongly associated with mortality among men (P = .01). Mortality was lower among women than among men, but morbidity was not consistently higher. The lack of gender differences in the association between morbidity and mortality suggests that this is not a likely explanation for the gender paradox, which refers to higher morbidity but lower mortality among women than among men.


[Restriction analysis of streptomyces plasmid pSS 181].
Restriction analysis of plasmid was carried out with the use of 7 enzymes. Presence of restriction sites for endonucleases II EcoRI, Pstl, BamHI, SalGI, and BgII was established. A unique restriction site for endonuclease EcoRI on the plasmid pSS181 was found out, but endonucleases HindIII and XbaI had no restriction sites on the plasmid. The molecular size of this plasmid was 10.75 +/- 0.25 kb. As a result of hydrolysis of the plasmid DNAby pairs ofendonucleases (EcoRI+PstI) and (EcoRI+BamHI) it was established, that the site of restriction for enzyme EcoRI was located on the biggest PstI and BamHI-fragments.


Eyelid myeloid sarcoma: ominous presentation of acute myelogenous leukemia.
A 19 year-old African American man presented to our clinic for a second opinion about a right upper eyelid mass which had been recalcitrant to treatment for nonspecific orbital inflammation by an outside facility. Examination for systemic causes of the patients eyelid lesion led to a diagnosis of acute myelogenous leukemia (AML) FAB subtype M1. A subsequent biopsy of the eyelid tumor confirmed the diagnosis of a myeloid sarcoma. The patient succumbed to complications from his leukemia within 13 months of presentation. This case report is the first, to our knowledge, of an eyelid myeloid sarcoma as the presenting sign of AML and demonstrates the poor prognosis of this lesion.


Castration-induced bone loss triggers growth of disseminated prostate cancer cells in bone.
Up to 90% of patients with castrate-resistant prostate cancer develop bone metastases, and the majority of these men have received androgen deprivation therapy known to cause bone loss. Whether this treatment-induced change to the bone microenvironment affects disseminated tumour cells, potentially stimulating development of bone metastasis, remains to be determined. The objective of this study was to use an in vivo model mimicking androgen ablation to establish the effects of this intervention on disseminated prostate cancer cells in bone. We mimicked the effects of androgen deprivation on bone metastasis by castrating 12-week-old BALB/c nude mice that had disseminated, hormone-insensitive PC3 prostate cancer cells present in the long bones. Castration caused increased bone resorption and loss of bone volume, compared with sham operation. In addition, castration triggered growth of disseminated PC3 cells to form bone metastasis in 70% of animals. In contrast, only 10% of sham-operated animals had detectable long bone tumours. Weekly administration of 100 μg/kg zoledronic acid (ZOL) prevented castration-induced tumour growth in bone and increased bone volume, but did not eliminate the disseminated tumour cells. ZOL had no effect on tumour growth in the sham-operated animals, despite causing a significant increase in bone volume. This is the first demonstration that, in a model of prostate cancer bone metastasis, mimicking androgen ablation results in growth of disseminated tumour cells in bone through osteoclast-mediated mechanisms. We provide the first biological evidence supporting the administration of ZOL to prostate cancer patients at the time of androgen ablation to prevent subsequent relapse in bone.


Maintenance of venous access devices in patients with neutropenia.
Neutropenia is a common complication in patients with cancer who are undergoing chemotherapy. Because of treatment frequency, potential chemotherapy damage to peripheral veins and tissues, and pain from multiple venipunctures, venous access devices (VADs) often are used in the oncology setting. Although VADs have been used for 30 years, no sufficient scientific data exist to support the best care and maintenance strategies for their use in patients with neutropenia. Understanding the etiology and pathogenesis of VAD-related infections (VAD-RI) and the proper steps to prevent or quickly treat it can decrease the likelihood that patients with neutropenia will have a fatal response. This article describes the differences between VADs, the most common sources of VAD-RI, and the treatment of such infections and suggests nursing interventions to help prevent VAD-RI.


Use of noninvasive fluorometry and spectrophotometry to study epithelial metabolism and transport.
Various examples illustrating the use of spectrophotometry and fluorometry in epithelia are presented. The first example uses the redox level of cytochrome aa3, measured spectrophotometrically as an index of tissue anoxia in cortical tubules and slices from the rabbit kidney. In the second example the redox level is used to measure the kinetics of aerobic energy production during transition to anoxia in the midgut of the tobacco hornworm. In the third application, the redox level of mitochondrial NADH is measured fluorometrically in a cortical tubule suspension from the rabbit kidney. Inhibition of active transport work causes reduction of NAD whereas increased work elicits oxidation of NAD, both occurring as expected from mitochondrial transitions to a lesser or more active state, respectively. Another use of NADH fluorescence is the determination of the relative effectiveness of metabolic substrates to deliver reducing equivalents to the respiratory chain in a particular tissue. Redox changes in mitochondrial NAD may be used to distinguish between primary metabolic and primary transport effects of hormones, drugs, and changes in the state of the organism. Finally, examples are provided of the use of an intracellular pH-sensitive dye and an extracellular calcium-sensitive dye in kidney tubules.


A Patient Portal Intervention for Menopause Knowledge and Shared Decision-Making.
Background: Menopause is a time often fraught with changes and symptoms, which may require difficult choices and decision-making. During this period, women would benefit from a better understanding and in-depth discussions with providers regarding menopause, associated conditions, and appropriate therapy. Patient portals offer a potential means to improve knowledge and shared decision-making (SDM) about menopause. Materials and Methods: This protocol article explores the feasibility of using the secure messaging (SM) function of the Veterans Affairs (VA) Patient Portal, "My HealtheVet" to implement an educational intervention and measure its impact on knowledge and SDM in the management of menopause. Results: This is a quality improvement pilot study in which women veterans of menopausal age in the Miami VA are offered an educational intervention via a patient portal, while women veterans in two neighboring VA facilities are not. Intervention participants receive weekly SMs with information on menopause symptoms, and treatment. After 6-months, all participants are surveyed on menopause knowledge, SDM, and satisfaction with the program. Conclusion: This study is among the first to assess the impact of an innovative patient portal intervention to improve knowledge and SDM between patients and providers regarding menopause. If successful, our program will add to the "meaningful use" of patient portals and offer a scalable and timely resource for SDM about menopause.


New Molecular Approach for the Detection of Kinetoplastida Parasites of Medical and Veterinary Interest.
Kinetoplastids are protozoa containing a range of ubiquitous free_living species-pathogens of invertebrates, vertebrates and even some plants. Some of them are causative agents of canine vector-borne diseases. Their diagnosis is often missing in a gold standard. Here, we proposed a molecular approach for the diagnosis and study of Kinetoplastida. The TaqMan qPCR assays target the following genes: 24Sa LSU of Kinetoplastida, 28S LSU of Leishmania/ Trypanosoma spp., 5.8S rRNA of Trypanosoma spp., 18S SSU of Leishmania spp., kinetoplast minicircle DNA (kDNA) of L. donovani complex and kDNA of L. infantum, were designed, validated for their sensitivity (Se) and specificity (Sp) in silico and in vitro using a panel of known DNAs. They were then used to screen 369 blood samples (358 dogs, 2 equids, 9 monkeys). In addition, new 28S LSU primer sets are presented to use for Kinetoplastida's identification by PCR/sequencing. All qPCRs showed consistently high analytical sensitivities and reproducibility. They detect approximately 0.01 parasite/ mL blood for the kDNA based- qPCRs and at least a single cell-equivalent of rDNA for the other systems. Based on the sequencing results, after screening, Se and Sp were: 0. 919 and 0.971, 0.853 and 0.979, 1.00 and 0.987, 0.826 and 0.995 for all of Kinetoplastida, Leishmania/ Trypanosoma, Trypanosoma, Leishmania spp. specific qPCRs, respectively. kDNA based qPCRs were more sensitive and specific (Se: 1.00; Sp: 0.997). PCR/sequencing allowed the detection of Kinetoplastids in animal blood samples such as L. infantum, L. guyanensis, T. congolense, T. evansi and Bodo spp. The molecular approach proposed here is useful for epidemiological studies, fundamental research such as screening for new Kinetoplastida species, diagnosis and therapeutic follow-up. In addition, researchers are free to choose the molecular tools adapted to their aims.


Multi-technique hybrid imaging in PET/CT and PET/MR: what does the future hold?
Integrated positron-emission tomography and computed tomography (PET/CT) is one of the most important imaging techniques to have emerged in oncological practice in the last decade. Hybrid imaging, in general, remains a rapidly growing field, not only in developing countries, but also in western industrialised healthcare systems. A great deal of technological development and research is focused on improving hybrid imaging technology further and introducing new techniques, e.g., integrated PET and magnetic resonance imaging (PET/MRI). Additionally, there are several new PET tracers on the horizon, which have the potential to broaden clinical applications in hybrid imaging for diagnosis as well as therapy. This article aims to highlight some of the major technical and clinical advances that are currently taking place in PET/CT and PET/MRI that will potentially maintain the position of hybrid techniques at the forefront of medical imaging technologies.


Surgical treatment of thumb carpometacarpal joint arthritis: a single institution experience from 1995-2005.
There are numerous techniques for the surgical management of thumb carpometacarpal (CMC) joint arthritis. The four senior authors of this study employ three such techniques: trapeziectomy with hematoma distraction arthroplasty, hemitrapeziectomy with osteochondral allograft, and ligament reconstruction tendon interposition (LRTI). This study examines the three commonly utilized procedures at a single institution. This study examines the 10-year experience from 1995-2005 with a minimum 3-month follow-up. Disabilities of the arm, shoulder, and hand (DASH) scores, pre-and postoperative pinch strength, and operative time were examined. After approval from the institutional review board of our institution was obtained, all patients treated surgically by three of the senior authors were contacted via mail and phone. Each patient was asked to complete and return a DASH questionnaire. Of the 115 patients treated during that period, 60 participated in this study. Each patient's final postoperative pinch measurement was obtained from occupational therapy and clinic records. This pinch strength was compared to the preoperative pinch and contralateral pinch strength. Lastly, the total operative time for each procedure was obtained from the operative record. The only significant finding in this study was a shorter mean operative time with the trapeziectomy group (76.90 min) and osteochondral allograft group (90.45 min) when compared to the LRTI group (139.00 min; p = 0.001 and p = 0.001, respectively). We found no significant difference between groups in terms of DASH score and pinch strength. There was no difference between the techniques in terms of postoperative pinch strength and patient satisfaction measured by DASH scores. The operative times for trapeziectomy and hematoma interposition as well as the osteochondral allograft were significantly shorter than that of the LRTI. This presents further evidence that potentially, "less is more" in the treatment of thumb CMC arthritis. We used a retrospective study design to evaluate potential differences between the three surgical techniques described above, therapeutic, levels III-IV.


Cerebrovascular/cardiovascular diseases and mental disorders due to overwork and work-related stress among local public employees in Japan.
In Japan, overwork-related disorders occur among local public employees as well as those in private businesses. However, to date, there are no studies reporting the state of compensation for cerebrovascular/cardiovascular diseases (CCVD) and mental disorders due to overwork or work-related stress among local public employees in Japan over multiple years. This report examined the recent trend of overwork-related CCVD and mental disorders, including the incidence rates of these disorders, among local public employees in Japan from the perspective of compensation for public accidents, using data from the Japanese Government and relevant organizations. Since 2000, compared to CCVD, there has been an overall increase in the number of claims and cases of compensation for mental disorders. Over half of the individuals receiving compensation for mental disorders were either in their 30s or younger. About 47% of cases of mental disorders were compensated due to work-related factors other than long working hours. The incidence rate by job type was highest among "police officials" and "fire department officials" for compensated CCVD and mental disorders cases, respectively. Changes in the trend of overwork-related disorders among local public employees in Japan under a legal foundation should be closely monitored.


Characterization of an insecticidal toxin and pathogenicity of Pseudomonas taiwanensis against insects.
Pseudomonas taiwanensis is a broad-host-range entomopathogenic bacterium that exhibits insecticidal activity toward agricultural pests Plutella xylostella, Spodoptera exigua, Spodoptera litura, Trichoplusia ni and Drosophila melanogaster. Oral infection with different concentrations (OD = 0.5 to 2) of wild-type P. taiwanensis resulted in insect mortality rates that were not significantly different (92.7%, 96.4% and 94.5%). The TccC protein, a component of the toxin complex (Tc), plays an essential role in the insecticidal activity of P. taiwanensis. The ΔtccC mutant strain of P. taiwanensis, which has a knockout mutation in the tccC gene, only induced 42.2% mortality in P. xylostella, even at a high bacterial dose (OD = 2.0). TccC protein was cleaved into two fragments, an N-terminal fragment containing an Rhs-like domain and a C-terminal fragment containing a Glt symporter domain and a TraT domain, which might contribute to antioxidative stress activity and defense against macrophagosis, respectively. Interestingly, the primary structure of the C-terminal region of TccC in P. taiwanensis is unique among pathogens. Membrane localization of the C-terminal fragment of TccC was proven by flow cytometry. Sonicated pellets of P. taiwanensis ΔtccC strain had lower toxicity against the Sf9 insect cell line and P. xylostella larvae than the wild type. We also found that infection of Sf9 and LD652Y-5d cell lines with P. taiwanensis induced apoptotic cell death. Further, natural oral infection by P. taiwanensis triggered expression of host programmed cell death-related genes JNK-2 and caspase-3.


Anti-citrullinated protein antibody titre as a predictor of abatacept treatment persistence in patients with rheumatoid arthritis: a prospective cohort study in Japan.
Objective: Successful rheumatoid arthritis (RA) outcome depends on treatment efficacy in the early stages of the disease and its sustainability. It is thus critical to identify factors predicting treatment persistence with biological agents, such as abatacept. We compared clinical profiles, including early changes in autoantibody titres at 3 months, between patients with RA demonstrating sustained persistence and those discontinuing abatacept treatment.Method: We prospectively enrolled 71 and 78 active RA patients treated with abatacept and tumour necrosis factor inhibitors (TNF-Is), respectively, who had previous disease-modifying anti-rheumatic drug) failure. Clinical characteristics were compared between non-continuation and continuation groups stratified according to abatacept or TNF-I persistence for at least 12 months from treatment initiation.Results: Significantly larger decreases in rheumatoid factor titre and anti-citrullinated protein autoantibody (ACPA) titre were observed in the continuation group of abatacept therapy at 3 months, and early reduction in ACPA titre remained a significant and independent predictor of sustained persistence with abatacept in multivariate analysis. In addition, we obtained the area under the receiver operator characteristics curve of 0.904 from a model including baseline ACPA titre and reduction of ACPA titre at 3 months. Sustained reduction of RA disease activity score at 12 months was significantly and independently associated with reduced ACPA titre at 3 months.Conclusions: Persistence with abatacept and sustained therapeutic response are associated with an early reduction in ACPA titre. Prediction of abatacept continuation and efficacy will facilitate the optimal design of therapy in the early stages of RA.


Protein intrinsic viscosity determination with the Viscosizer TD instrument: reaching beyond the initially expected applications.
Intrinsic viscosity is a key hydrodynamic parameter to understand molecular structure and hydration, as well as intramolecular interactions. Commercially available instruments measure intrinsic viscosity by recording the macromolecular mobility in a capillary. These instruments monitor Taylor dispersion using an absorbance or fluorescence detector. By design, these instruments behave like U-tube viscometers. To our knowledge, there are no studies to date showing that the Viscosizer TD instrument (Malvern-Panalytical) is able to measure the intrinsic viscosity of macromolecules. In this study, we then performed our assays on the Poly(ethylene oxide) polymer (PEO), used classically as a standard for viscometry measurements and on three model proteins: the bovine serum albumin (BSA), the bevacizumab monoclonal antibody, and the RTX Repeat Domain (RD) of the adenylate cyclase toxin of Bordetella pertussis (CyaA). The presence of P20 in the samples is critical to get reliable results. The data obtained with our in-house protocol show a strong correlation with intrinsic viscosity values obtained using conventional techniques. However, with respect to them, our measurements could be performed at relatively low concentrations, between 2 and 5 mg/ml, using only 7 µL per injection. Altogether, our results show that the Viscosizer TD instrument is able to measure intrinsic viscosities in a straightforward manner. This simple and innovative approach should give a new boost to intrinsic viscosity measurements and should reignite the interest of biophysicists, immunologists, structural biologists and other researchers for this key physicochemical parameter.


[The Melkersson-Rosenthal syndrome. Apropos 2 cases].
Melkersson-Rosenthal syndrome is classically defined as facial palsy, oedema facial and plication of the tongue, constituting one rare cause of facial palsy. The primary care physician must include this syndrome in the differential diagnosis of Bell's palsy. We report two cases of this syndrome, one in a young man, another in a 56 aged woman, and also a succinct literature review.


Incidence and outcomes of ventilator-associated pneumonia in Japanese intensive care units: the Japanese nosocomial infection surveillance system.
To determine the incidence of ventilator-associated pneumonia (VAP) among intensive care unit (ICU) patients in Japan and to assess the impact of VAP on patient outcomes, including mortality, length of stay, and duration of mechanical ventilation. Multicenter cohort study. Twenty-eight ICUs in multidisciplinary Japanese hospitals with more than 200 beds. A total of 21,909 patients 16 years or older who were admitted to an ICU between June 2002 and June 2004, stayed in the ICU for 24 to 1,000 hours, and were not transferred to another ICU. The overall infection rates for nosocomial pneumonia and VAP were 6.5 cases per 1,000 patient-days and 12.6 cases per 1,000 ventilator-days, respectively. The standardized mortality rates for the patients with VAP was 1.3 (95% confidence interval [CI], 1.1-1.6): 1.1 (95% CI, 0.9-1.4) for the cases due to drug-susceptible pathogens and 1.5 (95% CI, 1.1-1.9) for the cases due to drug-resistant pathogens. After adjusting for Acute Physiology and Chronic Health Evaluation II score, the mean length of stay for the patients with VAP caused by drug-susceptible pathogens (15.2 days [95% CI, 14.6-15.8]) and by drug-resistant pathogens (17.8 days [95% CI, 17.0-18.6]) was significantly longer than that in the patients without nosocomial infection (6.8 days [95% CI, 6.7-6.9]). The mean duration of mechanical ventilation in the patients with VAP caused by drug-susceptible pathogens (12.0 days [95% CI, 11.5-12.5]) and drug-resistant pathogens (14.1 days [95% CI, 13.5-14.8]) was significantly longer than that in the patients without nosocomial infection (4.7 days [95% CI, 4.6-4.8]). The incidence of VAP is substantial among ICU patients in Japan. The potential impact of VAP on patient outcomes emphasizes the importance of preventive measures against VAP, especially for VAP caused by drug-resistant pathogens.


Inductively Coupled Plasma Mass Spectrometry: Introduction to Analytical Aspects.
Inductively coupled plasma mass spectrometry (ICP-MS) is an analytical technique that can be used to measure elements at trace levels in biological fluids. Although older techniques such as atomic absorption and atomic emission are still in use by some laboratories, there has been a slow shift toward ICP-MS, particularly in the last decade. As this shift is likely to continue, clinical scientists should be aware of the analytical aspects of ICP-MS, as well as the potential for both spectroscopic and non-spectroscopic interference, and strategies that can be employed to eliminate or mitigate these issues.


How society should respond to the risk of vaccine rejection.
While vaccine acceptance remains high in general, fear of vaccines has grown dramatically in the past several years in many developed countries. In some communities, this fear has led to significantly increased rates of vaccine refusal which are associated with increases in illness and death from vaccine-preventable diseases, and large economic costs for health care and society. Despite overwhelming evidence supporting the safety and benefits of vaccination, this fear has proven resistant to information campaigns, a phenomenon well-explained by psychological research which has established that risk perception is subjective, a product of both the facts and how those facts feel. Given the innately emotional and instinctive nature of risk perception, and the risks to public health these perceptions produce, and consistent with well-established legal principles supporting government action to protect the common good, society has the right and responsibility to establish laws, regulations, and choice frameworks that discourage vaccine refusal.