Studying longitudinal neutralising antibody levels against Equid herpesvirus 1 in experimentally infected horses using a novel pseudotype based assay.
Infection with equid herpesvirus 1 (EHV-1), a DNA virus of the Herpesviridae family represents a significant welfare issue in horses and a great impact on the equine industry. During EHV-1 infection, entry of the virus into different cell types is complex due to the presence of twelve glycoproteins (GPs) on the viral envelope. To investigate virus entry mechanisms, specific combinations of GPs were pseudotyped onto lentiviral vectors. Pseudotyped virus (PV) particles bearing gB, gD, gH and gL were able to transduce several target cell lines (HEK293T/17, RK13, CHO-K1, FHK-Tcl3, MDCK I & II), demonstrating that these four EHV-1 glycoproteins are both essential and sufficient for cell entry. The successful generation of an EHV-1 PV permitted development of a PV neutralisation assay (PVNA). The efficacy of the PVNA was tested by measuring the level of neutralising serum antibodies from EHV-1 experimentally infected horses (n = 52) sampled in a longitudinal manner. The same sera were assessed using a conventional EHV-1 virus neutralisation (VN) assay, exhibiting a strong correlation (r = 0.82) between the two assays. Furthermore, PVs routinely require -80 °C for long term storage and a dry ice cold-chain during transport, which can impede dissemination and utilisation in other stakeholder laboratories. Consequently, lyophilisation of EHV-1 PVs was conducted to address this issue. PVs were lyophilised and pellets either reconstituted immediately or stored under various temperature conditions for different time periods. The recovery and functionality of these lyophilised PVs was compared with standard frozen aliquots in titration and neutralisation tests. Results indicated that lyophilisation could be used to stably preserve such complex herpesvirus pseudotypes, even after weeks of storage at room temperature, and that reconstituted EHV-1 PVs could be successfully employed in antibody neutralisation tests.


Issues in biometrics: offset potentials and the electrical stability of Ag/AgCl electrodes.
There has been a continuing interest in many areas of psychophysiological research in the quantification of slow wave bioelectric potentials recorded from the surface of the skin. However, surprisingly little research has focused specifically on methods for stabilizing and minimizing the offset potentials of the sintermetallic Ag/AgCl surface electrodes commonly used in such noninvasive bioelectric measurement. To provide empirically based recommendations in this regard, we contrasted the effects of three common storage methods (air dry vs. saline vs. carbon rod) on the two distinct types of Ag/AgCl surface electrodes (sintermetallic vs. electrolytic) under simulated experimental conditions. The obtained results confirm that minimum offset potentials and maximum stability for electrolytic Ag/AgCl electrodes are obtained when they are stored in a 0.9% NaCl solution with their leads shorted together and connected in parallel to a carbon rod partially immersed in the solution (cf. Cooper, 1956). In addition, the pattern of results suggested that the inter-session unshorted storage of sintermetallic electrodes in a mild saline solution is to be recommended over either shorted saline storage in parallel with a carbon rod or unshorted dry storage.


[Control of tuberculosis in refugees and displaced].
As a result of national and international conflicts, the number of refugees and displaced persons in various countries of the world is increasing. The complex and protracted nature of these conflicts often forces refugees to remain away from their countries for long periods, living in refugee camps. Many refugees come from countries where tuberculosis is endemic and, once the immediate problems of establishing a camp are overcome, this disease becomes the principal problem affecting refugee camps. In order to advise the persons in charge of refugee camps on how to set up tuberculosis control programs, this document details the general requirements and specific recommendations of WHO for the implementation of such programs, as well as guidelines for their evaluation and monitoring, including criteria that would justify camp closure.


[Modern medical problems of microcirculation and hypoxic syndrome].
In this paper long known problems of microcirculation are shown, which were solved only during the last 40 years. They are concerned with the velocity and character of the capillary blood flow, the regulation of the capillary blood flow, the role of various vessels in the oxygen transport, the role of leukocytes in physiology and pathology of the capillary blood flow, with the special features of the function of lungs in supplying the whole organism with oxygen and with bioenergetic laws in the development of an organism adaptation to hypoxia. Here we considered a number of the most important medical problems of microcirculation and hypoxic syndrome. A relatively new factor in the capillary circulation is the fact that in the brain and heart capillaries there are sites with pO2 close to zero. They show that the capillary circulation has no central nervous regulation of the blood flow. The blood flow in these organs obeys only occasional oscillations. The new fact is that Krogh's rule about metabolism and oxygen exchange occurring only in the capillaries is abandoned. It is shown that almost 30% of consumed oxygen is delivered to the brain via arterioles, which changes our relation to the capillary circulation as a unique mechanism of the tissue supply with oxygen. The new fact is also the mass adhesion of leukocytes to the walls of microvessels, which results in the occlusion of the vessels followed by the development of the heart and brain ischemia. It was shown for the first time that contrary to previous ideas the alveoli in the lungs are supplied with blood from a powerful network of large microvessels from 20 to 50 microm in diameter rather than from thin arterioles. They make possible the passage of 6-121 of the blood in the norm and during stressed muscle activity--up to 18-231 of blood per minute. The principle is substantiated that during hypoxia only normal supply of an organism with oxygen may result in a complete adaptation of an organism to the deficit of oxygen.


Alpha-1 Antitrypsin PiMZ Genotype Is Associated with Chronic Obstructive Pulmonary Disease in Two Racial Groups.
Alpha-1 antitrypsin deficiency, caused primarily by homozygosity for the Z allele of the SERPINA1 gene, is a well-established genetic cause of chronic obstructive pulmonary disease (COPD). Whether the heterozygous PiMZ genotype for alpha-1 antitrypsin confers increased risk for COPD has been debated. We analyzed 8,271 subjects in the Genetic Epidemiology of COPD (COPDGene) Study, hypothesizing that PiMZ would independently associate with COPD and COPD-related phenotypes. The COPDGene Study comprises a multiethnic, cross-sectional, observational cohort of non-Hispanic white and African American current and former smokers with at least 10 pack-years of smoking who were enrolled for detailed clinical and genetic studies of COPD and COPD-related traits. We performed multivariate logistic regression analysis for moderate to severe COPD and assessed Pi genotype with other relevant covariates in models stratified by race. We analyzed quantitative characteristics on the basis of volumetric computed tomography with generalized linear models controlling for genotype, scanner type, and similar covariates. White PiMZ COPDGene subjects had significantly lower lung function, FEV1 percent predicted (68 ± 28 vs. 75 ± 27; P = 0.0005), and FEV1/FVC ratio (0.59 ± 0.18 vs. 0.63 ± 0.17; P = 0.0008), as well as more radiographic emphysema (P = 0.001), than subjects without alpha-1 antitrypsin Z risk alleles. Similarly, African American PiMZ subjects had lower lung function, FEV1 percent predicted (65 ± 33 vs. 84 ± 25; P = 0.009) and FEV1/FVC (0.61 ± 0.21 vs. 0.71 ± 0.15; P = 0.03). In the COPDGene Study, we demonstrate that PiMZ heterozygous individuals who smoke are at increased risk for COPD and obstructive lung function impairment compared with Z-allele noncarriers, regardless of race. Although severe alpha-1 antitrypsin deficiency is uncommon in African Americans, our study adds further support for initial targeted detection of all subjects with COPD for alpha-1 antitrypsin deficiency, including African Americans. Clinical trial registered with www.clinicaltrials.gov (NCT00608784).


Domestic wastewater treatment by a submerged MBR (membrane bio-reactor) with enhanced air sparging.
The air sparging technique has been recognised as an effective way to control membrane fouling. However, its application to a submerged MBR (Membrane Bio-Reactor) has not yet been reported. This paper deals with the performances of air sparging on a submerged MBR for wastewater treatment. Two kinds of air sparging techniques were used respectively. First, air is injected into the membrane tube channels so that mixed liquor can circulate in the bioreactor (air-lift mode). Second, a periodic air-jet into the membrane tube is introduced (air-jet mode). Their applicability was evaluated with a series of lab-scale experiments using domestic wastewater. The flux increased from 23 to 33 l m(-2) h(-1) (43% enhancement) when air was injected for the air-lift module. But further increase of flux was not observed as the gas flow increased. The Rc/(Rc+Rf), ratio of cake resistance (Rc) to sum of Rc and Rf (internal fouling resistance), was 23%, indicating that the Rc is not the predominant resistance unlike other MBR studies. It showed that the cake layer was removed sufficiently due to the air injection. Thus, an increase of airflow could not affect the flux performance. The air-jet module suffered from a clogging problem with accumulated sludge inside the lumen. Because the air-jet module has characteristics of dead end filtration, a periodic air-jet was not enough to blast all the accumulated sludge out. But flux was greater than in the air-lift module if the clogging was prevented by an appropriate cleaning regime such as periodical backwashing.


Association of PDGF-BB-induced thrombomodulin with the regulation of inflammation in the corneal and scleral stroma.
The responses of corneal and scleral stromal cells to platelet-derived growth factor (PDGF)-BB were assessed and inflammatory reactions in the cornea and sclera were investigated. Primary cultures of cells obtained from human subjects and strains derived from human corneal or scleral stromal cells (Cs3 and Sc1, respectively) were used. Changes in gene expression after 24 hours of exposure to 10 ng/mL PDGF-BB were analyzed with an Sc1 DNA microarray. The upregulation of several genes in Cs3 and Sc1 was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. The expression of bioactive factors was detected immunohistochemically in nine different clinical specimens. DNA microarray analysis revealed that the gene encoding thrombomodulin (TM) was induced in Sc1 by PDGF-BB. RT-PCR confirmed that TM expression at the mRNA level was increased in both corneal and scleral stromal cells. At the protein level, TM expression was increased in scleral stromal cells, but not in corneal cells, and TM was detected in both the membrane and cytoplasmic compartments. TM was detected immunohistochemically in inflamed scleral and several corneal specimens. After TM stimulation, interleukin (IL)-18 transcription was increased in Sc1. PDGF-BB induced TM mRNA expression in scleral and corneal stromal cells, but Western blot analysis revealed the increase in TM expression only in the scleral cells. TM induced IL-18 in scleral stromal cells. A cascade involving these biologically active factors may regulate scleral and corneal inflammation. The results also reveal differences in the biological response of scleral and corneal stromal cells.


The digital hearing aid, wearable computing, and electrophysiological response.
Presbycusis is the most common form of hearing loss caused by aging and long-term exposure to sound energy. This type of ailment decreases the ear's ability to perceive high frequencies and localize sound, thus making comprehension more difficult. To compensate for this loss, the choice of a digital hearing aid has become more common. However, most digital hearing aids do little more that their analogue predecessor's by providing a single, fixed hearing response. Such a fixed response is not suitable for all of a user's auditory environments and typically causes a more rapid loss of hearing. Significant advancement in processing power and reduction in size of computing hardware has produced increasingly more powerful, more portable, and more personal computing devices. These advances have spurred research and development of wearable computing devices towards integrating both man and machine. By definition, the digital hearing aid is a wearable computing device. The development of a digital hearing aid with increased onboard processing that is aware of its owner's electrophysiological and auditory environments is an obvious progression. This awareness will give the hearing aid the ability to autonomously modify its own parameters to improve audibility and comprehension. Electrophysiological signals can be classified as naturally occurring or voluntarily controlled. Employing these signals will allow the hearing aid to adapt to its owner's external and internal stimuli. Research and initial experiments into the monitoring and use of electrophysiological response for the control of the digital hearing aid shall be presented.


Store-operated Ca(2+) entry is expressed in human endothelial progenitor cells.
Endothelial progenitor cells (EPCs) may be recruited from the bone marrow to sites of tissue regeneration to sustain neovascularization and reendothelialization after acute vascular injury. This feature makes them particularly suitable for cell-based therapy. In mature endothelium, store-operated Ca(2+) entry (SOCE) is activated following emptying of inositol-1,4,5-trisphosphate-sensitive stores, and controls a wide number of functions, including proliferation, nitric oxide synthesis, and vascular permeability. The present work aimed at investigating SOCE expression in EPCs harvested from both peripheral blood (PB-EPCs) and umbilical cord blood (UCB-EPCs) by employing both Ca(2+) imaging and molecular biology techniques. SOCE was induced upon either pharmacological (ie, cyclopiazonic acid) or physiological (ie, ATP) depletion of the intracellular Ca(2+) pool. Further, store-dependent Ca(2+) entry was inhibited by the SOCE inhibitor, N-(4-[3,5-bis(trifluoromethyl)-1H-pyrazol-1-yl]phenyl)-4-methyl-1,2,3-thiadiazole-5-carboxamide (BTP-2). Real-time reverse transcription-polymerase chain reaction and western blot analyses showed that both PB-EPCs and UCB-EPCs express all the molecular candidates to mediate SOCE in differentiated cells, including TRPC1, TRPC4, Orai1, and Stim1. Moreover, pharmacological maneuvers demonstrated that, as well as in differentiated endothelial cells, the signal transduction pathway leading to depletion of the intracellular Ca(2+) pool impinged on the phospholipase C/inositol-1,4,5-trisphosphate pathway. Finally, blockage of SOCE with BTP-2 impaired PB-EPC proliferation. These findings provide the first evidence that EPCs express SOCE, which might thus be regarded as a novel target to enhance the regenerative outcome of cell-based therapy.


False-negative core needle biopsies of the breast: an analysis of clinical, radiologic, and pathologic findings in 27 concecutive cases of missed breast cancer.
A benign diagnosis in a core needle biopsy (CNBx) of the breast performed for a clinically and/or radiologically suspicious abnormality is often due to a nonrepresentative sample. However, the discordance may not be recognized, resulting in a logistic delay in the diagnosis. Twenty-seven false-negative CNBxs were identified in 952 consecutive CNBxs of the breast (653 benign, 266 malignant, and 33 atypical) performed during a 1-year period. Biopsies were analyzed with respect to clinical and radiologic findings, biopsy type, type of malignancy, and interval between the original CNBx and final diagnosis. Four hundred thirty-eight (67%) of the patients with a benign CNBx diagnosis either underwent excision or had a minimum of 1-year follow-up (mean, 35.6 months; median, 36 months). The cancers missed on CNBx included 6 ductal carcinomas in situ, 17 invasive ductal carcinomas, 3 invasive lobular carcinomas, and 1 non-Hodgkin lymphoma. The overall false-negative rate was 9.1%. For palpable lesions, ultrasound-guided CNBx had a lower rate of missed cancer (3.6%) compared with CNBx without image guidance (13.3%). The false-negative rate for vacuum assisted CNBx biopsy was 7.6% (3.3% for the 11-gauge needle, 22.2% for the 14-gauge needle; 5.6% for nonpalpable mass lesions, 8.2% for microcalcifications). In all seven false-negative CNBxs performed by radiologists, the discordance between the radiologic and pathologic findings was promptly recognized due to their standard follow-up protocol. The discordance between the degree of clinical suspicion, radiologic impression, and the pathologic findings was not immediately recognized in 5 of 20 false-negative CNBxs performed by surgeons (4 without radiologic guidance and 1 with ultrasound guidance), resulting in a delay in the diagnosis ranging from 112-336 days. A false-negative diagnosis of breast carcinoma was found to be more common in CNBx performed without image guidance but occurred to a lesser degree in image-guided biopsies. A delay in diagnosis can be avoided by establishing a standard post-CNBx follow-up protocol.


Early Events in Coccidioidomycosis.
Since its description nearly 130 years ago, hundreds of studies have deepened our understanding of coccidioidomycosis, also known as valley fever (VF), and provided useful diagnostic tests and treatments for the disease caused by the dimorphic fungi Coccidioides spp. In general, most of the literature has addressed well-established infections and has described patients who have experienced major complications. In contrast, little attention has been given to the earliest consequences of the pathogen-host interaction and its implications for disease manifestation, progression, and resolution. The purpose of this review is to highlight published studies on early coccidioidomycosis, identify gaps in our knowledge, and suggest new or former research areas that might be or remain fertile ground for insight into the early stages of this invasive fungal disease.


Benchmarking RNA-Seq quantification tools.
RNA-Seq, a deep sequencing technique, promises to be a potential successor to microarrays for studying the transcriptome. One of many aspects of transcriptomics that are of interest to researchers is gene expression estimation. With rapid development in RNA-Seq, there are numerous tools available to estimate gene expression, each producing different results. However, we do not know which of these tools produces the most accurate gene expression estimates. In this study we have addressed this issue using Cufflinks, IsoEM, HTSeq, and RSEM to quantify RNA-Seq expression profiles. Comparing results of these quantification tools, we observe that RNA-Seq relative expression estimates correlate with RT-qPCR measurements in the range of 0.85 to 0.89, with HTSeq exhibiting the highest correlation. But, in terms of root-mean-square deviation of RNA-Seq relative expression estimates from RT-qPCR measurements, we find HTSeq to produce the greatest deviation. Therefore, we conclude that, though Cufflinks, RSEM, and IsoEM might not correlate as well as HTSeq with RT-qPCR measurements, they may produce expression values with higher accuracy.


Driver impairment at night and its relation to physiological sleepiness.
Studies of devices detecting sleepiness need reference points of physiological sleepiness. The present study sought to validate the Karolinska drowsiness score (KDS) as an indicator of physiological sleepiness against driving impairment and eye blink duration during a 45-minute drive in an advanced moving-base driving simulator. Data from 19 persons were used in the analysis. Electrooculography, electroencephalography, and electromyography were administered continuously. Physiological sleepiness was quantified by scoring the percentage (0-100%) of the scoring epoch with alpha and theta activity and slow eye movements (KDS). Lateral position and speed were used as measures of driving behavior. Lane departure was defined as two wheels touching the lane markers. Blink duration was used as a secondary indicator of sleepiness. The results showed that, for young drivers, sleepiness increased with time in the task with higher levels. The variability of the lateral position and the mean and variability of the blink duration significantly changed when sleepiness increased to KDS > or =20%. Furthermore, there was an increase in the risk of lane departure for KDS > or =30%. The results suggest that KDS scoring is a reasonable procedure for estimating physiological sleepiness under conditions of driving. The results also indicate that a younger age is associated with greater sensitivity to sleepiness at the wheel.


Plasma Chemerin Is Induced in Critically Ill Patients with Gram-Positive Infections.
Chemerin is a chemoattractant protein abundantly expressed in hepatocytes. Chemerin exerts pro- and anti-inflammatory effects and acts as a pro-resolving protein. Chemerin levels are low in patients with liver cirrhosis and are increased in sepsis. The aim of this study was to identify associations between plasma chemerin levels and underlying diseases as well as causes of severe illness. The cohort included 32 patients with liver cirrhosis who had low systemic chemerin, and who were not considered for further evaluation. Plasma chemerin levels were similar between the 27 patients with systemic inflammatory response syndrome (SIRS), the 34 patients with sepsis and the 63 patients with septic shock. Chemerin in plasma correlated with C-reactive protein and leukocyte count but not with procalcitonin, a clinical marker of bacterial infection. Plasma chemerin did not differ among patients with and without ventilation and patients with and without dialysis. Vasopressor therapy was not associated with altered plasma chemerin levels. Infection with severe acute respiratory syndrome coronavirus 2 had no effect on plasma chemerin levels. Baseline levels of plasma chemerin could not discriminate between survivors and non-survivors. Notably, Gram-positive infection was associated with higher chemerin levels. In summary, the current study suggests that plasma chemerin might serve as an early biomarker for the diagnosis of Gram-positive infections in patients with sepsis.


Attributes of conscientiousness throughout the animal kingdom: An empirical and evolutionary overview.
Conscientiousness is a fundamental aspect of human personality, one that is closely linked with various favorable life outcomes. Despite its importance in humans, conscientiousness has received little attention as to how it may have evolved, or whether it provides similar fitness benefits in other animals. To date, research in animal personality has found consistent support for the presence of all major dimensions of human personality in other animals except conscientiousness. In this review, we investigate conscientiousness at the level of traits and facets (clusters of closely related traits). A systematic review of the literature retrieved 876 relevant publications describing attributes of conscientiousness in other animal species. A factor analysis of citation counts revealed 2 major dimensions representing 9 distinct facets of conscientiousness in nonhumans. These facets, together with their underlying personality traits, exhibit individual variability, are generally known to be heritable, and often offer clear fitness benefits to individuals. Other facets of conscientiousness appear to be unique to humans. Publication biases, research biases, and anthropomorphism may all play a role in the structure of the evidence we report. We conclude by suggesting fruitful areas of future research to further elucidate the presence and functional roles of conscientiousness in animals. (PsycINFO Database Record


Alterations in conserved Kir channel-PIP2 interactions underlie channelopathies.
Inwardly rectifying K(+) (Kir) channels are important regulators of resting membrane potential and cell excitability. The activity of Kir channels is critically dependent on the integrity of channel interactions with phosphatidylinositol 4,5-bisphosphate (PIP(2)). Here we identify and characterize channel-PIP(2) interactions that are conserved among Kir family members. We find basic residues that interact with PIP(2), two of which have been associated with Andersen's and Bartter's syndromes. We show that several naturally occurring mutants decrease channel-PIP(2) interactions, leading to disease.


Leaching- and sintering-resistant hollow or structurally ordered intermetallic PtFe alloy catalysts for oxygen reduction reactions.
Carbon supported Pt-based alloy materials that have been developed for proton exchange membrane fuel cells (PEMFCs) are vulnerable to deactivation due to the loss of non-noble metal components (leaching) or detachment, migration and aggregation of active nanoparticles (sintering). Until now some methods have been developed to inhibit leaching or sintering individually. However, a route able to avoid leaching and sintering simultaneously is still lacking. Herein, we develop a thermally driven interfacial diffusion alloying route that allows for the direct evolution of solid Pt nanoparticles (NPs) supported on carbon (Pt/C) into a Pt-skin-like hollow PtFe alloy or a structurally ordered intermetallic PtFe alloy, together with in situ encapsulation of PtFe alloy NPs with a thin layer porous nitrogen-doped carbon (NC) shell. The robust NC shells not only effectively prevent Pt-based NPs from detachment, migration, and aggregation during accelerated durability tests but also allow smoother access of electrolyte to the Pt surface, thus allowing the catalysts to well preserve their high catalytic activity. The well-defined shape and atomic arrangement of PtFe alloy NPs exhibit over 600% increase in mass activity and specific activity when compared with that of the pristine Pt/C catalyst. Stability tests confirm that the ordered PtFe alloy is more electrochemically stable than the disordered hollow PtFe alloy and Pt/C catalysts due to its ordered atomic arrangement and the robust NC shell.


Protocol adherence for continuously titrated interventions in randomized trials: an overview of the current methodology and case study.
The standard definition for protocol adherence is the proportion of all scheduled doses that are delivered. In clinical research, this definition has several limitations when evaluating protocol adherence in trials that study interventions requiring continuous titration. Building upon a specific case study, we analyzed a recent trial of a continuously titrated intervention to assess the impact of different definitions of protocol deviations on the interpretation of protocol adherence. The OVATION pilot trial was an open-label randomized controlled trial of higher (75-80 mmHg) versus lower (60-65 mmHg) mean arterial pressure (MAP) targets for vasopressor therapy in shock. In this trial, potential protocol deviations were defined as MAP values outside the targeted range for >4 consecutive hours during vasopressor therapy without synchronous and consistent adjustments of vasopressor doses. An adjudication committee reviewed each potential deviation to determine if it was clinically-justified or not. There are four reasons for this contextual measurement and reporting of protocol adherence. First, between-arm separation is a robust measure of adherence to complex protocols. Second, adherence assessed by protocol deviations varies in function of the definition of deviations and the frequency of measurements. Third, distinguishing clinically-justified vs. not clinically-justified protocol deviations acknowledges clinically sensible bedside decision-making and offers a clear terminology before the trial begins. Finally, multiple metrics exist to report protocol deviations, which provides different information but complementary information on protocol adherence. In trials of interventions requiring continuous titration, metrics used for defining protocol deviations have a considerable impact on the interpretation of protocol adherence. Definitions for protocol deviations should be prespecified and correlated with between-arm separation, if it can be measured.


Decreased expression of the insulin-like growth factor 1 receptor by ribozyme cleavage.
Insulin-like growth factor (IGF)-1 and its receptor (IGF-1R) are associated with abnormal retinal neovascularization. Ribozymes were designed that selectively decreased the expression of the IGF-1R and these ribozymes were tested in angiogenesis models in vitro and in vivo. Two hammerhead ribozymes were designed that cleave the human IGF-1R mRNA. The ribozymes were cloned into recombinant adeno-associated viral vectors (rAAV). The rAAV constructs were transfected into human retinal endothelial cells (HRECs). IGF-1R mRNA and protein levels were examined and the modified Boyden chamber assay used to examine ribozyme effects on cell migration. These constructs were injected intravitreally into mice to determine the effect of the ribozymes on retinal neovascularization in a mouse model of oxygen-induced retinopathy. Relative quantitative RT-PCR analysis showed that IGF-1R Rz1 reduced IGF-1R mRNA levels by 40% +/- 10% (P = 0.003), and Western blot analysis showed a 41% +/- 5% (P = 4.6 x 10(-5)) reduction of IGF-1R protein, confirming that this ribozyme reduces IGF-1R expression. IGF-1R Rz1 also reduced IGF-1-induced cell migration by 90% +/- 5% (P = 2.9 x 10(-9)) showing that IGF-1R Rz1 reduces IGF-1R function in HRECs. IGF-1R Rz1 also reduced the amount of preretinal neovascularization by 65% +/- 6% (P = 2.7 x 10(-5)), as measured by the average number of endothelial preretinal nuclei per section. These studies demonstrate that the IGF-1R ribozymes are effective at reducing the expression and function of the IGF-1R in vitro and in vivo. Therefore, the IGF-1R ribozymes are an effective method for studying the process of angiogenesis and may ultimately be effective as gene therapy tools for the reduction of pathologic retinal angiogenesis.


Conversion of Bacillus subtilis 168 to a subtilin producer by competence transformation.
Subtilin is a ribosomally synthesized peptide antibiotic produced by Bacillus subtilis ATCC 6633. B. subtilis 168 was converted to a subtilin producer by competence transformation with chromosomal DNA from B. subtilis ATCC 6633. A chloramphenicol acetyltransferase gene was inserted next to the subtilin structural gene as a selectable marker. The genes that conferred subtilin production were derived from a 40-kb region of the B. subtilis ATCC 6633 chromosome that had flanking homologies to the B. subtilis 168 chromosome. The subtilin produced by the mutant was identical to natural subtilin in its biological activity, chromatographic behavior, amino acid composition, and N-terminal amino acid sequence.


Effect of acute exposure to low environmental calcium on respiration and locomotion in Lymnaea stagnalis (L.).
Environmental calcium is a major factor affecting the distribution of freshwater gastropods. Whilst the effects on growth and morphology are fairly well understood, little is known about how calcium availability affects other aspects of gastropod biology. Lymnaea stagnalis (L.) is considered a calciphile and exhibits reduced growth and survival in environments containing less than 20 mg l(-1) Ca(2+). Many freshwater systems exhibit fluctuations in calcium concentration over time: where calcium levels are normally high there may be periods of low [Ca(2+)], for example following periods of flooding. Here we examined the effects of acute periods of low (20 mg l(-1)) environmental calcium on the physiology and behaviour of L. stagnalis, specifically measuring how locomotion and respiration differ between high calcium (80 mg l(-1)) and low calcium (20 mg l(-1)) environments. We found that in a low calcium environment crawling speed is reduced, and that this coincides with an increase in cutaneous respiration, indicating that the increased metabolic demands of calcium acquisition at low [Ca(2+)] reduce the energy available for locomotion. Conversely we found a decrease in aerial respiration in hypoxic conditions in the low calcium relative to the high calcium environment. In conclusion, we found that acute exposure to low environmental calcium has a highly significant effect on locomotion and respiration, which may have consequences for snail fitness when no morphological effects are apparent.


Olive Leaf Extract (OleaVita) Suppresses Inflammatory Cytokine Production and NLRP3 Inflammasomes in Human Placenta.
The placenta is essential for pregnancy and produces both pro-inflammatory and anti-inflammatory cytokines. Excessive production of inflammatory cytokines, involving interleukin-1β (IL-1β), IL-6, and IL-8, from placental tissues is associated with pregnancy complications. Olive leaf extract has several health benefits, including anti-inflammatory functions. OleaVita is a new commercial olive leaf extract; it is hypothesized to suppress placental inflammation. In human placental tissue culture, OleaVita treatment inhibited the secretion of inflammatory cytokines and NF-κB p65 protein expression. OleaVita also suppressed toll-like receptor ligands-induced IL-1β secretion in human placental tissues. IL-1β is regulated by the NLRP3 inflammasomes, a pivotal regulator of various diseases. OleaVita significantly decreased NLRP3 and pro-IL-1β protein expression, suggesting that it has an inhibitory effect on NLRP3 inflammasome activation. Thus, OleaVita is beneficial as an inhibitor of inflammation and NLRP3 inflammasome activation, and may be used as a supplement for the treatment and prevention of inflammatory diseases.


Barriers to and facilitators of independent non-medical prescribing in clinical practice: a mixed-methods systematic review.
What are the factors that affect the implementation or utilisation of independent non-medical prescribing (iNMP)? Mixed-methods systematic review. Two reviewers independently completed searches, eligibility and quality assessments. Pre-defined search terms were utilised to search electronic databases. Reference lists, key journals and grey literature were searched alongside consultation with authors/experts. Qualitative and quantitative studies investigating independent prescribing by any non-medical professional group. Study participants included any stakeholders involved in actual or proposed iNMP. Measurements reported on data describing stakeholders' perceptions and experiences of the barriers to/facilitators of iNMP. A total of 43 qualitative and seven quantitative studies from three countries (n=12, 117 participants) were included. Quality scores varied from 9 to 35 (Quality Assessment Tool for Studies with Diverse Designs, 0 to 48). Qualitative data were synthesised into four themes (and subthemes): systems (government and political, organisational, formulary); education and support (non-medical prescribing (NMP) courses/continuous professional development (CPD)); personal and professional (medical profession, NMP professions, service users); and financial factors. Quantitative data corroborated the qualitative themes. Integration of the qualitative themes and quantitative data enabled the development of a NMP implementation framework. Barriers to and facilitators of the implementation and utilisation of iNMP are evident, demonstrating multifactorial and context-specific variables within four explicit themes. Professional bodies, politicians, policy and healthcare managers and clinicians could use the resulting NMP implementation framework to ensure the safe and successful implementation and utilisation of NMP. Clinical physiotherapists and other clinicians should consider whether these variables have been adequately addressed prior to adopting NMP into their clinical practice. PROSPERO CRD42015017212. [Noblet T, Marriott J, Graham-Clarke E, Rushton A (2017) Barriers to and facilitators of independent non-medical prescribing in clinical practice: a mixed-methods systematic review. Journal of Physiotherapy 63: 221-234].


STED microscopy reveals dendrite-specificity of spines in turtle cortex.
Dendritic spines are key structures for neural communication, learning and memory. Spine size and shape probably reflect synaptic strength and learning. Imaging with superresolution STED microscopy the detailed shape of the majority of the spines of individual neurons in turtle cortex (Trachemys scripta elegans) revealed several distinguishable shape classes. Dendritic spines of a given class were not distributed randomly, but rather decorated significantly more often some dendrites than others. The individuality of dendrites was corroborated by significant inter-dendrite differences in other parameters such as spine density and length. In addition, many spines were branched or possessed spinules. These findings may have implications for the role of individual dendrites in this cortex.


Risk-adapted survival benefit of IMRT in early-stage NKTCL: a multicenter study from the China Lymphoma Collaborative Group.
This study evaluated the survival benefit of intensity-modulated radiation therapy (IMRT) compared with 3-dimension conformal radiation therapy (3D-CRT) in a large national cohort of patients with early-stage extranodal nasal-type natural killer/T-cell lymphoma (NKTCL). This retrospective study reviewed patients with early-stage NKTCL treated with high-dose radiation therapy (RT; ≥45 Gy) at 16 Chinese institutions. Patients were stratified into 1 of 4 risk groups based on the number of risk factors: low risk (no factors), intermediate-low risk (1 factor), intermediate-high risk (2 factors), and high-risk (3-5 factors). Of the 1691 patients, 981 (58%) received IMRT, and 710 (42%) received 3D-CRT. Unadjusted 5-year overall survival (OS) and progression-free survival (PFS) were 75.9% and 67.6%, respectively, for IMRT compared with 68.9% (P = .004) and 58.2% (P < .001), respectively, for 3D-CRT. After propensity score match and multivariable analyses to account for confounding factors, IMRT remained significantly associated with improved OS and PFS. The OS and PFS benefits of IMRT persisted in patients treated with modern chemotherapy regimens. Compared with 3D-CRT, IMRT significantly improved OS and PFS for high-risk and intermediate-high-risk patients but provided limited benefits for low-risk or intermediate-low-risk patients. A risk-adapted survival benefit profile of IMRT can be used to select patients and make treatment decisions.


Assessing the psychometric properties of the World Assumptions Scale.
The notion that traumatic events shatter survivors' fundamental assumptions about the world has been accepted as a truism in the trauma literature, although tests of this hypothesis provide a somewhat mixed picture. Because this research has relied heavily on the World Assumptions Scale (WAS), it is unclear whether these inconsistent results are due to problems with assumptive world theory, improper tests of the theory, or problems with the WAS. Data were gathered from 742 undergraduate students at four universities to assess the psychometric properties of the WAS. Findings suggest several shortcomings of the WAS, including temporal instability, and mixed evidence regarding construct validity. Implications for trauma research are discussed.


Neural-network models for the blood glucose metabolism of a diabetic.
We study the application of neural networks to modeling the blood glucose metabolism of a diabetic. In particular we consider recurrent neural networks and time series convolution neural networks which we compare to linear models and to nonlinear compartment models. We include a linear error model to take into account the uncertainty in the system and for handling missing blood glucose observations. Our results indicate that best performance can be achieved by the combination of the recurrent neural network and the linear error model.


Unraveling the RNA modification code with mass spectrometry.
The discovery and analysis of modifications on proteins and nucleic acids has provided functional information that has rapidly accelerated the field of epigenetics. While protein post-translational modifications (PTMs), especially on histones, have been highlighted as critical components of epigenetics, the post-transcriptional modification of RNA has been a subject of more recently emergent interest. Multiple RNA modifications have been known to be present in tRNA and rRNA since the 1960s, but the exploration of mRNA, small RNA, and inducible tRNA modifications remains nascent. Sequencing-based methods have been essential to the field by creating the first epitranscriptome maps of m6A, m5C, hm5C, pseudouridine, and inosine; however, these methods possess significant limitations. Here, we discuss the past, present, and future of the application of mass spectrometry (MS) to the study of RNA modifications.


Mechanistic investigation of food effect on disintegration and dissolution of BCS class III compound solid formulations: the importance of viscosity.
A negative food effect, i.e. a decrease in bioavailability upon the co-administration of compounds together with food, has been attributed particularly with high solubility/low permeability compounds (BCS class III). Different mechanisms have been proposed including intestinal dilution leading to a lower concentration gradient across the intestinal wall as well as binding of the active pharmaceutical ingredient to food components in the intestine and thereby decreasing the fraction of the dose available for absorption. These mechanisms refer primarily to the compound and not to the dosage form. An increase in viscosity of the dissolution fluid will in particular affect the absorption of BCS type III compounds with preferential absorption in the upper small intestine if the API release is delayed from the dosage form. The present study demonstrated that the increase in viscosity of the dissolution medium, following ingestion of a solid meal, may drastically reduce disintegration and dissolution. For that purpose the viscosity of the standard FDA meal was determined and simulated by solutions of HPMC in buffer. As model formulations, three commercially available tablets containing trospium chloride, a BCS class III m-cholinoreceptor antagonist was used. Trospium chloride drug products have been described to undergo a negative food effect of more than 80% following ingestion with food. The tablets showed prolonged disintegration times and reduced dissolution rates in viscous media, which could be attributed to changes in the liquid penetration rates. The effect was particularly significant for film-coated tablets relative to uncoated dosage forms. The results show the necessity of considering media viscosity when designing in vitro models of drug release for BCS type III drug formulations.


Navigating Challenging Conversations: The Interplay Between Inquiry and Knowledge Drives Preparation for Future Learning.
While some physicians hone their skills through informal learning in clinical practice, others do not. There is a lack of understanding of why some physicians seek improvement and how they use the workplace context to build their capabilities. Because physicians rarely pursue formal professional development activities to improve communication skills, examining physician-patient communication offers a powerful opportunity to illuminate important aspects of preparation for future learning in the workplace. This qualitative observational study involved over 100 hours of observation of eight pediatric rehabilitation physicians as they interacted with patients and families at an academic teaching hospital in 2018-2020. Detailed field notes of observations, post-observation interviews, and exit interviews were the data sources. Data collection and analysis using a constructivist grounded theory approach occurred iteratively, and themes were identified through constant comparative analysis. Through their daily work, experienced physicians employ 'habits of inquiry' by constantly seeking a better understanding of how to navigate challenging conversations in practice through monitoring and attuning to situational and contextual cues, taking risks and navigating uncertainty while exploring new and varied ways of practicing, and seeking why their strategies are successful or not. Engaging in communication challenges drives physician learning through an interplay between habits of inquiry and knowledge: inquiry into how to improve their communication supported by existing conceptual knowledge to generate new strategies. These 'habits of inquiry' prompt continual reinvestment in problem solving to refine existing knowledge and to build new skills for navigating communication challenges in practice.


Optimal posture control of a musculo-skeletal arm model.
In this paper maximal performance posture control of the human arm is investigated by means of model simulations. Recent experiments (F.C.T. van der Helm, submitted, 2000) have shown that the reflexive feedback during postural control varies with the bandwidth of the applied force disturbances. This paper focusses on the influence of the frequency content of force disturbances on the reflexive feedback gains by means of optimization. The arm is modelled by a nonlinear musculo-skeletal model with two degrees of freedom and six muscles. To facilitate the optimization of the model parameters, the arm model is linearized. A performance criterion is minimized for stochastic force disturbances in a two-step procedure: (1) optimization of static muscle activations using an additional energy criterion to obtain a unique and energy-efficient solution; antid (2) optimization of reflex gains using an additional control effort criterion to obtain a unique solution. The optimization reveals that for the given task and posture, the shoulder muscles have the largest contribution, whereas the bi-articular muscles have a relatively small contribution to the behaviour. The dynamics at the endpoint level are estimated so that a comparison can be made with the experiments. Compared to the experiments, the intrinsic damping of the model is relatively large (about 150%), whereas the intrinsic stiffness is relatively small (about 60%). These differences can be attributed to unmodelled mechanical effects of crossbridges in Hill-type muscle models. The optimized reflex gains show remarkable similarities with the values found in the experiments, implying that humans can adjust their reflexive feedback gains in an optimal way, weighting the performance and energy. The approach in this paper could be useful in the study of various posture tasks, for example in the prediction of the relation between the control parameters of various musculo-skeletal models and different experimental variables.


Calcium entry through receptor-operated channels in bovine pulmonary artery endothelial cells.
The activation of endothelial cells by endothelium-dependent vasodilators has been investigated using bioassay, patch clamp and 45Ca flux methods. Cultured pulmonary artery endothelial cells have been demonstrated to release EDRF in response to thrombin, bradykinin, ATP and the calcium ionophore A23187. The resting membrane potential of the endothelial cells was -56 mV and the cells were depolarized by increasing extracellular K+ or by the addition of (0.1-1.0 mM)Ba2+ to the bathing solution. The electrophysiological properties of the cultured endothelial cells suggest that the membrane potential is maintained by an inward rectifying K+ channel with a mean single channel conductance of 35.6 pS. The absence of a depolarization-activated inward current and the reduction of 45Ca influx with high K+ solution suggests that there are no functional voltage-dependent calcium or sodium channels. Thrombin and bradykinin were shown to evoke not only an inward current (carried by Na+ and Ca2+) but also an increase in 45Ca influx suggesting that the increase in intracellular calcium necessary for EDRF release is mediated by an opening of a receptor operated channel. High doses of thrombin and bradykinin induced intracellular calcium release, however, at low doses of thrombin no intracellular calcium release was observed. We propose that the increased cytosolic calcium concentration in endothelial cells induced by endothelium dependent vasodilators is due to the influx of Ca2+ through a receptor operated ion channel and to a lesser degree to intracellular release of calcium from a yet undefined intracellular store.


Retention and release of oil-in-water emulsions from filled hydrogel beads composed of calcium alginate: impact of emulsifier type and pH.
Delivery systems based on filled hydrogel particles (microgels) can be fabricated from natural food-grade lipids and biopolymers. The potential for controlling release characteristics by modulating the electrostatic interactions between emulsifier-coated lipid droplets and the biopolymer matrix within hydrogel particles was investigated. A multistage procedure was used to fabricate calcium alginate beads filled with lipid droplets stabilized by non-ionic, cationic, anionic, or zwitterionic emulsifiers. Oil-in-water emulsions stabilized by Tween 60, DTAB, SDS, or whey protein were prepared by microfluidization, mixed with various alginate solutions, and then microgels were formed by simple extrusion into calcium solutions. The microgels were placed into a series of buffer solutions with different pH values (2 to 11). Lipid droplets remained encapsulated under acidic and neutral conditions, but were released under highly basic conditions (pH 11) due to hydrogel swelling when the alginate concentration was sufficiently high. Lipid droplet release increased with decreasing alginate concentration, which could be attributed to an increase in the pore size of the hydrogel matrix. These results have important implications for the design of delivery systems to entrap and control the release of lipophilic bioactive components within filled hydrogel particles.


Factors precipitating outbreaks of measles in district Kangra of North India: A case-control study.
Globally, measles is the fifth killer disease among children under five years of age. Despite high immunization coverage in Himachal, outbreaks are occurring. Upon two outbreaks in a hilly district in North India, a case control study was conducted to identify factors contributing to outbreaks and to recommend remedial measures to prevent further outbreaks. FACTORS WERE REVIEWED UNDER THREE HEADS: program related, health care providers, and beneficiaries related. Cold chain maintenance was determined and responses were compared between workers from study Shahpur and control Nagrota Bagwan blocks. All 69 mothers of age and sex matched children with measles were enrolled. A pre-designed pre-tested data collection instrument was used. For statistical analysis, the odds ratio (OR) and adjusted odds ratio with 95% confidence interval (C.I.) among women of children exposed and unexposed to selected characteristics were calculated. Poor cold chain maintenance and gaps in knowledge of health workers supplemented with beneficiary-related issues precipitated outbreaks in case area. Univariate analysis yielded strong statistical significance to 17 variables. Important statistically significant variables are educational status; OR 27.63 (C.I. 9.46-85.16); occupation; OR 0.35 (C.I. 0.16-0.75); income; OR 5.49 (C.I. 2.36-13.00); mode of transport to health care facility; OR 8.74 (C.I. 2.90-28.23); spread of illness from one person to another; OR 5.60 (95% C.I. 1.40-25.97); first help for sick child OR 2.12 (C.I. 1.00-4.50), and place of visit after recovery; OR 3.92 (C.I. 1.80-8.63). Multiple logistic regression yielded significant association with educational status, drinking water sources, and time taken to reach the nearest health facility. Measles outbreaks were confirmed in high immunization coverage areas. We recommend 2nd dose opportunity for measles (MR) between 5 and 17 years; refresher trainings to workers; mobile access to health care facility, and Information Education Communication activities for social behavioral change in affected areas.


Development of an optimised key worker framework for people with dementia, their family and caring unit living in the community.
In Australia, dementia is a national health priority. With the rising number of people living with dementia and shortage of formal and informal carers predicted in the near future, developing approaches to coordinating services in quality-focused ways is considered an urgent priority. Key worker support models are one approach that have been used to assist people living with dementia and their caring unit coordinate services and navigate service systems; however, there is limited literature outlining comprehensive frameworks for the implementation of community dementia key worker roles in practice. In this paper an optimised key worker framework for people with dementia, their family and caring unit living in the community is developed and presented. A number of processes were undertaken to inform the development of a co-designed optimised key worker framework: an expert working and reference group; a systematic review of the literature; and a qualitative evaluation of 14 dementia key worker models operating in Australia involving 14 interviews with organisation managers, 19 with key workers and 15 with people living with dementia and/or their caring unit. Data from the systematic review and evaluation of dementia key worker models were analysed by the researchers and the expert working and reference group using a constant comparative approach to define the essential components of the optimised framework. The developed framework consisted of four main components: overarching philosophies; organisational context; role definition; and key worker competencies. A number of more clearly defined sub-themes sat under each component. Reflected in the framework is the complexity of the dementia journey and the difficulty in trying to develop a 'one size fits all' approach. This co-designed study led to the development of an evidence based framework which outlines a comprehensive synthesis of components viewed as being essential to the implementation of a dementia key worker model of care in the community. The framework was informed and endorsed by people living with dementia and their caring unit, key workers, managers, Australian industry experts, policy makers and researchers. An evaluation of its effectiveness and relevance for practice within the dementia care space is required.


No genetic association between polymorphisms in the Tau gene and Alzheimer's disease in clinic or population based samples.
Mutations in the tau protein gene have recently been found to cause familial fronto-temporal dementia in a number of kindreds demonstrating linkage to chromosome 17. Given that tau pathology is a hallmark of Alzheimer's disease (AD), this raises the possibility that mutations in tau may also be associated with AD. We have investigated the allelic frequencies of polymorphisms in the Tau gene for a possible allelic distortion in Alzheimer's cases, which might suggest a conferred genetic risk. We have genotyped 65 community-based and 200 clinic-based AD cases, and 142 community-based controls at the Tau exon 6 AflIII and BslI polymorphisms and find no independent association with risk for AD in these samples. Further analysis including APOE genotypes from the same samples demonstrated no interaction between either of these polymorphisms and APOE in conferring risk for AD. In addition, haplotype analysis across both sites revealed no difference in haplotype frequencies between cases and controls, nor any interaction with APOE. Therefore our data do not suggest any association between these variations in the Tau gene and Alzheimer's disease.


Cryo-EM structures of PP2A:B55-FAM122A and PP2A:B55-ARPP19.
Progression through the cell cycle is controlled by regulated and abrupt changes in phosphorylation.1 Mitotic entry is initiated by increased phosphorylation of mitotic proteins, a process driven by kinases,2 while mitotic exit is achieved by counteracting dephosphorylation, a process driven by phosphatases, especially PP2A:B55.3 While the role of kinases in mitotic entry is well-established, recent data have shown that mitosis is only successfully initiated when the counterbalancing phosphatases are also inhibited.4 For PP2A:B55, inhibition is achieved by the two intrinsically disordered proteins (IDPs), ARPP19 (phosphorylation-dependent)6,7 and FAM122A5 (inhibition is phosphorylation-independent). Despite their critical roles in mitosis, the mechanisms by which they achieve PP2A:B55 inhibition is unknown. Here, we report the cryo-electron microscopy structures of PP2A:B55 bound to phosphorylated ARPP19 and FAM122A. Consistent with our complementary NMR spectroscopy studies both IDPs bind PP2A:B55, but do so in highly distinct manners, unexpectedly leveraging multiple distinct binding sites on B55. Our extensive structural, biophysical and biochemical data explain how substrates and inhibitors are recruited to PP2A:B55 and provides a molecular roadmap for the development of therapeutic interventions for PP2A:B55 related diseases.


Human population structure and its effects on sampling Y chromosome sequence variation.
The excess of rare variants in global sequencing studies of the nonrecombining portion of the Y chromosome (NRY) has been interpreted as evidence for the effects of human demographic expansion. However, many NRY polymorphisms are geographically localized and the effect of different geographical sampling on patterns of NRY variation is unknown. We use two sampling designs to detect population structure and its effects on patterns of human NRY polymorphism. First, we sequence 26.5 kb of noncoding Y chromosome DNA from 92 globally distributed males representing 35 populations. We find that the number of polymorphisms with singleton variants is positively correlated with the number of populations sampled and that there is a significant negative correlation of Tajima's D (TD) and Fu and Li's D (FD) statistics with the number of pooled populations. We then sequence the same region in a total of 73 males sampled from 3 distinct populations and find that TD and FD values for the 3 pooled and individual population samples were much less negative than those in the aforementioned global sample. Coalescent simulations show that a simple splitting model of population structure, with no changes in population size, is sufficient to produce the negative values of TD seen in our pooled samples. These empirical and simulation results suggest that observed levels of NRY population structure may lead to an upward bias in the number of singleton variants in global surveys and call into question inferences of population expansion based on global sampling strategies.


Isolation and characterization of a penicillinase from Pseudomonas cepacia 249.
Pseudomonas cepacia has an inducible beta-lactamase which is responsible for its novel ability to catabolize beta-lactam compounds. The gene encoding this enzyme, penA, was cloned from a genomic library of P. cepacia 249 on the broad-host-range cosmid pLAFR. This separated the penA gene from the gene encoding a second beta-lactamase in P. cepacia 249. Expression of penA was inducible in an Escherichia coli host strain by low levels of penicillin. The 33,500-molecular-weight enzyme had penicillinase activity not inhibited by clavulanic acid or sulbactam and was highly active against piperacillin and azlocillin. In comparison with other inducible beta-lactamases produced by gram-negative organisms, the penA enzyme had many properties which were similar to those of the penicillinase produced by Alcaligenes faecalis. It was unlike the ampC-type cephalosporinase produced by Pseudomonas aeruginosa.


Roles of the H-2D(b) and H-K(b) genes in resistance to persistent Theiler's murine encephalomyelitis virus infection of the central nervous system.
Theiler's murine encephalomyelitis virus, a member of the Picornaviridae family, persists in the spinal cord of susceptible strains of mice. Resistant strains of mice, such as the H-2(b) strain, clear the virus infection after an acute encephalomyelitis. The H-2D locus, but not the H-2K locus, has a major effect on this resistance, although both loci code for MHC class I molecules with similar general properties. For the present work, we rendered susceptible H-2(q) FVB/N mice transgenic for either the H-2D(b)gene, the H-2K(b) gene or a chimeric H-2D(b)/K(b) gene in which the exons encoding the peptide-binding groove of the H-2K(b) gene have been replaced by those of the H-2D(b)gene. Mice transgenic for either the H-2D(b)gene or the chimeric H-2D(b)/K(b) gene were significantly more resistant to persistent virus infection than mice transgenic for the H-2K(b) gene, suggesting that the difference in the effects of the H-2D(b)gene and the H-2K(b) gene are due to the nature of the peptides presented by these class I molecules.


[Clinico-hemodynamic effects of psychotropic preparations and psychosomatic correlations in cardiac rhythm disorders].
A total of 136 patients with paroxysmal supraventricular tachyarrhythmia and tachyform of atrial fibrillation and polytopic extrasystoles were examined. The combination of cardiovascular and psychovegetative disturbances closely related to the expression of psychopathological shifts was revealed. The antiarrhythmic activity of psychotropic drugs (phenibut, sulpiride, pipofezin, tofizopam and falilepsin) depending on the nature of the time course of the emotional-personality status in prolonged intermittent treatment of patients with arrhythmias of the functional and mixed genesis was shown. A negative inotropic effect of phenibut, a hyperdynamic effect of sulpiride, pipofezin, falilepsin and a modulating effect of tofizopam were revealed. Prolonged therapy with psychopharmacological drugs in short courses was proposed.


Structural and Biochemical Changes in Pericardium upon Genipin Cross-Linking Investigated Using Nondestructive and Label-Free Imaging Techniques.
Tissue cross-linking represents an important and often used technique to enhance the mechanical properties of biomaterials. For the first time, we investigated biochemical and structural properties of genipin (GE) cross-linked equine pericardium (EP) using optical imaging techniques in tandem with quantitative atomic force microscopy (AFM). EP was cross-linked with GE at 37 °C, and its biochemical and biomechanical properties were observed at various time points up to 24 h. GE cross-linked EP was monitored by the normalized ratio between its second-harmonic generation (SHG) and two-photon autofluorescence emissions and remained unchanged for untreated EP; however, a decreasing ratio due to depleted SHG and elevated autofluorescence and a fluorescence band at 625 nm were found for GE cross-linked EP. The mean autofluorescence lifetime of GE cross-linked EP also decreased. The biochemical signature of GE cross-linker and shift in collagen bands were detected and quantified using shifted excitation Raman difference spectroscopy as an innovative approach for tackling artifacts with high fluorescence backgrounds. AFM images indicated a higher and increasing Young's modulus correlated with cross-linking, as well as collagen structural changes in GE cross-linked EP, qualitatively explaining the observed decrease in the second-harmonic signal. In conclusion, we obtained detailed information about the biochemical, structural, and biomechanical effects of GE cross-linked EP using a unique combination of optical and force microscopy techniques in a nondestructive and label-free manner.


Applied studies of the plant meridian system: II. Agri-wave technology increases the yield and quality of spinach and lettuce and enhances the disease resistant properties of spinach.
Agri-wave technology is composed of both a special frequency sound wave and a microelement fertilizer. In both components, the effect of sound waves on plants is more than that of fertilizers, but the best function is a combination of the two. Treatment by Agri-wave technology stimulated the growth rate and increased the yield of spinach. In small plot tests, the length and width of the treated spinach leaf was 50.8 cm and 20.3 cm, respectively, whereas the untreated leaves were 29.20 cm and 8.9 cm. The fresh weight of treated spinach was 0.42 kg. This was 5.5 times higher than that of the untreated spinach. In large area testing (17 hectares), the results of two tests show that the yields of the treated spinach were increased 22.7% and 22.2% over those of the control group. Sugar content of the treated spinach was increased by 37.5%, vitamin A, C, and B were increased 35.63%, 41.67% and 40.00%, respectively, above the levels of the control group. Niacin content was decreased by 7.69%. Of 33 elements analyzed in the spinach, 29 elements were increased by Agri-wave technology. The spinach was infected with "rot disease" in the control group while there was no disease present in the treated group. In greenhouse testing, the average weight of 3 species of lettuce treated by Agri-wave technology was increased 44.10% over that of the control group (P < 0.0001). The average weight of 3 species of lettuce by only sound and only fertilizer treated separately increased 29.92% and 16.19% above that of the control group (P < 0.0001). Sampling survey results in the field test were comparable to the above mentioned greenhouse test. The fresh weight of treated lettuce by Agri-wave technology was increased 41.67% over that of the control group (P < 0.0001). The fresh weight of treated lettuce by only sound and only fertilizer was increased 30.88% and 19.61%, respectively, over the control group (both P < 0.0001).


Phagocytosis of apoptotic eosinophils but not neutrophils by bronchial epithelial cells.
We have previously demonstrated that human bronchial epithelial cells engulf apoptotic eosinophils. To compare and contrast the phagocytic capabilities of monocyte-derived macrophage and primary airway epithelial cells for apoptotic granulocytes. Here we compared phagocytosis of human apoptotic eosinophils and neutrophils by small and large airway epithelial cells (SAEC and LAEC) and monocyte-derived macrophages. Confocal microscopy of F-actin staining and scanning and transmission electron microscopy revealed phagocytic cup formation around apoptotic eosinophils by airway epithelial cells (AEC) membranes with evidence of their digestion. Resting and cytokine-stimulated AEC did not recognize and ingest apoptotic neutrophils. The latter were phagocytosed by macrophages that exhibited greater ingestion of and higher capacity for, apoptotic eosinophils over apoptotic neutrophils. Cytochalasin D completely abolished uptake of apoptotic eosinophils by SAEC, LAEC or macrophage monolayers. Ligation of epithelial cell CD44 receptors for 24 h increased phagocytosis of apoptotic eosinophils by SAEC and LAEC with a potency comparable with that of IL-1. Phagocytosis was a specific receptor-mediated process involving integrin- (alphavbeta3, alphavbeta5, CD36), phosphatidylserine receptor- and lectin-dependent mechanisms. No significant differences were observed in avarice for apoptotic eosinophils by SAEC or LAEC either resting, CD44 monoclonal antibodies- or cytokine- stimulated, or in their usage and expression of recognition receptors. These findings further suggest and define an important role for the bronchial epithelium in the selective removal of apoptotic eosinophils from the airways in asthma.


[Work capacity of mitral stenosis patients before and after mitral commissurotomy].
Analysis of the working capacity of 1,000 patients who underwent mitral commissurotomy showed that 67.0% of individuals who were operated on for grade III stenosis and 27.99% of those operated on for grade IV stenosis resumed work. Frequent exacerbations of rheumatism and changes in the myocardium and valvular apparatus are among the causes of the decrease in the number of persons who resume work after the operation. Another reason is overestimation of the group of invalidity or its unjustified qualification


[First aid and prognosis following drowning accidents. Results of a retrospective study of 115 cases].
In 115 cases of submersion the initial findings of the rescue team, the patients status in the emergency room and the course of clinical treatment were analyzed retrospectively. Submersion accidents happened preferably in February, March and in the summertime from May to August. Most of the accidents took place in public waters or public baths (85.2%). Children below 10 years of age were involved in 34.8% of the submersion accidents. 57 patients were near drowned and 58 patients were drowned. The prognosis of patients with detectable heartbeat at the site of the accident depends on the primary pulmonary lesion. If respiratory insufficiency is recognized early and treated aggressively by intubation and mechanical ventilation with PEEP, these patients have an excellent prognosis. Only one patient with detectable heartbeat died, typically, after delayed treatment of respiratory failure. 55 patients recovered completely; one patient was suffering from a lesion of the n. medianus. Contrariwise, the prognosis of patients without detectable heartbeat is mainly determined by the consequences of hypoxaemia and is, overall, poor. Though resuscitation succeeds in 50% of submersion victims, only one out of four successfully resuscitated patients survived with little or no neurologic damage. Severe hypothermia may improve the prognosis of submersion victims. Thus, there are no useful parameters that would accurately predict the individual course of a submersion victim.


A full-spectral Bayesian reconstruction approach based on the material decomposition model applied in dual-energy computed tomography.
Dual-energy computed tomography (DECT) makes it possible to get two fractions of basis materials without segmentation. One is the soft-tissue equivalent water fraction and the other is the hard-matter equivalent bone fraction. Practical DECT measurements are usually obtained with polychromatic x-ray beams. Existing reconstruction approaches based on linear forward models without counting the beam polychromaticity fail to estimate the correct decomposition fractions and result in beam-hardening artifacts (BHA). The existing BHA correction approaches either need to refer to calibration measurements or suffer from the noise amplification caused by the negative-log preprocessing and the ill-conditioned water and bone separation problem. To overcome these problems, statistical DECT reconstruction approaches based on nonlinear forward models counting the beam polychromaticity show great potential for giving accurate fraction images. This work proposes a full-spectral Bayesian reconstruction approach which allows the reconstruction of high quality fraction images from ordinary polychromatic measurements. This approach is based on a Gaussian noise model with unknown variance assigned directly to the projections without taking negative-log. Referring to Bayesian inferences, the decomposition fractions and observation variance are estimated by using the joint maximum a posteriori (MAP) estimation method. Subject to an adaptive prior model assigned to the variance, the joint estimation problem is then simplified into a single estimation problem. It transforms the joint MAP estimation problem into a minimization problem with a nonquadratic cost function. To solve it, the use of a monotone conjugate gradient algorithm with suboptimal descent steps is proposed. The performance of the proposed approach is analyzed with both simulated and experimental data. The results show that the proposed Bayesian approach is robust to noise and materials. It is also necessary to have the accurate spectrum information about the source-detector system. When dealing with experimental data, the spectrum can be predicted by a Monte Carlo simulator. For the materials between water and bone, less than 5% separation errors are observed on the estimated decomposition fractions. The proposed approach is a statistical reconstruction approach based on a nonlinear forward model counting the full beam polychromaticity and applied directly to the projections without taking negative-log. Compared to the approaches based on linear forward models and the BHA correction approaches, it has advantages in noise robustness and reconstruction accuracy.


1-2 Drinks Per Day Affect Lipoprotein Composition after 3 Weeks-Results from a Cross-Over Pilot Intervention Trial in Healthy Adults Using Nuclear Magnetic Resonance-Measured Lipoproteins and Apolipoproteins.
Alcohol consumption ranging from 1−2 drinks/day associates with a lower risk of coronary heart disease in some studies. The underlying mechanisms are unclear. The Metabolic Imprints of Alcoholic Beverages (MetAl) trial aimed to explore the short-term effects of moderate alcohol consumption on cardiovascular biomarkers. A 2 × 3-week cross-over single-blinded intervention trial investigating the effect of 1−2 drinks/day (~12−24 g) compared with abstention on 1H Nuclear Magnetic Resonance-measured main lipoproteins and subfractions was performed in 26 healthy adults. Volunteers were classified as occasional or habitual drinkers based on their habitual alcohol intakes (<2 or ≥2 drinks/week). Compared with abstention, 1−2 drinks/day increased HDL2a-C (p = 0.004), HDL3-C (p = 0.008), and HDL non-significantly (p = 0.19). Total apoA1 and apoA1 in HDL and its subfractions increased (p < 0.05). Novel findings were a decreased apoB/apoA1 ratio (p = 0.02), and increased HDL2a phospholipid content (p = 0.04). In women alone, the results were similar but attenuated, and LDL-P decreased. Thus, changes in apoA1- and HDL-related biomarkers occur within weeks in moderate drinkers. Compared with abstention, 1−2 drinks/day increased total apoA1 more strongly than HDL-C and increased the cholesterol, apoA1, and phospholipid content of several HDL subfractions. Whether this provides a cardiovascular benefit requires further study. Clinicaltrials.gov: NCT03384147.


Detection of minor myocardial injury after successful percutaneous transluminal coronary angioplasty with or without stenting.
Cardiac troponins are sensitive and specific markers for the detection of minor myocardial injury. However, they have been rarely used to monitor myocardial injury after coronary stenting. The purpose of the study was to measure cardiac troponin I (cTnI) and cardiac troponin T (cTnT) levels after apparently successful percutaneous transluminal coronary angioplasty (PTCA) with or without coronary stenting and to compare their results with serum creatine kinase (CK) and its isoform, creatine kinase-MB (CKMB). CTnI and cTnT levels were compared with those of CK or CKMB in 50 consecutive patients with stable angina undergoing visually successful PTCA with stenting (n = 35) or without stenting (n = 15). Cardiac TnI, cTnT, CK and CKMB levels were measured before and 6, 24, and 48 hours after the procedures was performed. None of the patients had abnormal cTnI or cTnT levels, CK activity, or CKMB levels before the procedures. Moreover, no patient showed electrocardiographic evidence of myocardial infarction. 13 patients (26%) had abnormal peak values of one or more markers at 24 hours after coronary intervention. Troponin I was elevated in 10/35 patients after coronary stenting (29%) and in 2/15 patients after PTCA (13%) (P = 0.327). Troponin T was elevated in 6 patients (17%) and CKMB activity was elevated in 3 patients (9%) of the coronary stenting group. CTnI was more significant than CKMB (P = 0.023) in detecting minor myocardial injury. When compared with cTnI and CKMB, cTnT did not reach significance (P = 0.129 and 0.489, respectively). 5 out of the 13 patients with abnormal markers (38%) developed side branch occlusion after stenting. In conclusion, cTnI was a very sensitive marker in detecting minor myocardial injury after coronary angioplasty with or without stenting. The frequency of increased serum levels of cardiac troponins was higher in patients undergoing stent implantation than in those treated with angioplasty alone but did not reach significance. Side branch occlusion may have accounted for some, but not all, periprocedural minor myocardial injury in the stent group.


A Stimulus Artifact Removal Technique for SEMG Signal Processing During Functional Electrical Stimulation.
The purpose of this study was to design a method for extracting the volitional EMG from recorded surface electromyography (EMG), contaminated by functional electrical stimulation (FES) artifact. Considering that the FES artifact emerges periodically with rather large amplitude in nonstationary EMG, we designed an adaptive-matched filter (AMF) via genetic algorithm (GA) optimization. Both the simulated and real data from seven subjects were processed, using the GA-AMF filter and comb filter, respectively. To test the filtering effect on the EMG, contaminated with FES artifact of different current intensities, the contaminated EMG was simulated by combining the simulation artifact and clean EMG with various FES artifacts to clean EMG ratios. The results show that, in simulation test, compared to the EMG filtered by comb filter, the simulated EMG (p < 0.05), filtered by using GA-AMF, had significantly higher correlation coefficient, higher signal to noise ratio, and lower normalized root mean square error, whereas the real EMG (p < 0.05), filtered by using GA-AMF had higher power reduction than that filtered by using comb filter. The results indicate that GA-AMF can effectively remove FES artifact from the EMG of the stimulated muscle and its adjacent muscle, and the GA-AMF filter performed better than did the comb filter. All these results demonstrate that the GA-AMF filter is capable of extracting volitional EMG from the stimulated muscle and adjacent muscles. GA-AMF could provide technical support for improving EMG feedback control of FES rehabilitation system.


Products and kinetics of the heterogeneous reaction of particulate ametryn with NO3 radicals.
As a renowned s-triazine herbicide, ametryn is worldwide emitted into the atmosphere in both gaseous and particulate phases via spray drifts from treatments and post application emissions, but its chemical degradation in the atmosphere has not been well characterized. In this study, the heterogeneous kinetics of particulate ametryn with NO3 radicals were investigated with a mixed-phase relative rate method. A vacuum ultraviolet photoionization aerosol time-of-flight mass spectrometer (VUV-ATOFMS) and an atmospheric gas analysis mass spectrometer were synchronously used to online monitor the decays of particulate ametryn and gas-phase isoprene. The reactive uptake coefficient of NO3 radicals on ametryn particles was calculated to be 2.9 × 10(-2), according to the measured ametryn loss ratio and the average NO3 concentration. The effective rate constant for the heterogeneous reaction of particulate ametryn with NO3 radicals measured under experimental conditions was 8.4 × 10(-13) cm(3) molecule(-1) s(-1). In addition, atraton, ametryn sulfoxide and ametryn sulfone were identified as the reaction products by gas-chromatography-mass spectrometry (GC-MS) analysis. The experimental results might shed light on the chemical behavior of atmospheric ametryn at night-time.


Nationwide 2.5-Year School-Based Public Health Intervention Program Designed to Reduce the Incidence of Caries in Children of Grenada.
This paper describes an innovative public health intervention, called 'Smile Grenada', targeting the oral health of children in Grenada utilizing the resources of a US dental school, several oral health care companies, local governmental and public health authorities, and Grenadian school personnel. Preintervention visual/tactile caries examinations were collected from 1,092 schoolchildren (mean age 9.9 years, standard deviation, SD = 3.7) in 2010. The intervention included: (1) classroom-based toothbrushing with fluoridated toothpaste, (2) fluoride varnish applied by trained dental students, teachers and local providers 3 times a year and (3) glass ionomer sealants placed on first permanent molars in children aged 6-8 years. Postintervention data were collected in May, 2013 (n = 2,301, mean age 9.8 years, SD = 3.7). Decayed and demineralized surfaces were examined for the whole sample and decay/demineralization and sealant retention on 6-year molars were examined separately (ages 6-8 in 2013 cohort). The number of decayed/demineralized surfaces declined across all age groups. The average number of decayed surfaces dropped from 9 at baseline to just over 6 (F1, 3,393 = 69.8, p < 0.0001) and the average number of demineralized surfaces dropped from 6 to less than 2 (1.8 surfaces; F1, 3,393 = 819.0, p < 0.0001). For children aged 6-8 years, there were statistically significantly fewer decayed surfaces (t1, 2,086 = 12.40, p < 0.0001; mean baseline 0.93, SD = 1.75; mean follow-up 0.23, SD = 0.83) and demineralized surfaces (t1, 2,086 = 19.7, p < 0.0001; mean baseline 2.11, SD = 2.74; mean follow-up 0.50, SD = 0.97) on 6-year molars. The Smile Grenada program successfully demonstrated a locally sustainable model for improving oral health in children in a developing country.


Pharmacological treatments of cerebellar ataxia.
The confirmed pharmacological treatment of cerebellar ataxia is still lacking. In a recent preliminary trial, we showed that D-cycloserine, a partial NMDA allosteric agonist, may relieve the symptoms. In this paper, major clinical trials to relieve ataxic symptoms are reviewed. Previous studies showed some efficacy of physostigmine in ataxic patients. However, physostigmine did not improve the ataxia in a recent double-blind crossover study. The replacement therapy of the deficient cholinergic system with choline or choline derivatives was tried in patients with Friedreich's ataxia and other ataxic patients, but the result was not definitive. A levorotatory form of hydroxytryptophan (a serotonin precursor), a serotoninergic 5-HT1A agonist, a serotoninergic 5-HT3 antagonist, and a serotonin reuptake inhibitor were also used for the therapy for ataxia. In a double-blind randomized study, buspirone, a 5-HT1A agonist was active in cerebellar ataxia, but the effect is partial and not major. The effects of the studies with the other serotoninergic drugs were not consistent. The effect of sulfamethoxazole-trimethoprim therapy in spinocerebellar ataxia type3/Machado-Joseph disease (MJD) was reported, although the therapy improved spasticity or rigidity, rather than ataxia. In contrast to previous studies, sulfamethoxazole-trimethoprim therapy in MJD had no effect in a 2001 double-blind crossover study. The thyrotropin-releasing hormone, D-cycloserine, and acetazolamide for SCA6 may have some efficacy. However, a well-designed double-blind crossover trial is needed to confirm the effect.


Equilibria amongst different molybdenum (V)-containing species from sulphite oxidase. Evidence for a halide ligand of molybdenum in the low-pH species.
The interaction of chloride, fluoride and phosphate ions with the molybdenum centre of sulphite oxidase in the pH range 6.2 to 9.6 has been studied by e.p.r. of Mo(V) in the enzyme reduced by sulphite. Detailed studies were made from e.p.r. spectra recorded at about 120K and more limited studies from spectra of liquid samples at about 295K and also from enzyme activity measurements. Interconversion between low-pH and high-pH Mo(V) e.p.r. signal-giving species [described by Lamy, Gutteridge & Bray (1980) Biochem. J. 185, 397-403] is influenced by chloride concentration, a 10-fold increase in concentration (in the range of about 1 mM to 100 mM) causing an increase of about 1 pH unit in the apparent pK for the conversion. This suggests that chloride is a constituent of the low-pH species. In support of this, high concentrations of fluoride modified the e.p.r. spectrum. Partial conversion to a Mo(V) species, in which F- has presumably replaced Cl- and showing hyperfine coupling of A(19F)av. 0.5mT, is indicated. It is proposed that interconversion between high-pH and low-pH species is of the form: (formula; see text) No evidence that Cl- is essential for enzymic activity was found. Data relating to equilibria amongst low-pH, high-pH and also the phosphate species are presented. Depending on pH and on concentrations of Cl- and H2PO4-, one, two, or all three species may be present. Qualitatively, under appropriate conditions, the phosphate species tends to replace some or all of the low-pH species. Quantitative analysis by a computer procedure permitted an appropriate scheme to be deduced and equilibrium constants to be evaluated. Studies on the e.p.r. signals at 295K indicated that similar equilibria applied in liquid solution, but with some changes in the values of the constants. The structure of the molybdenum centre in its various states and the nature of the enzymic reaction are discussed.


Genetic analysis of mate discrimination in Drosophila simulans.
Courtship is an elaborate behavior that conveys information about the identity of animal species and suitability of individual males as mates. In Drosophila, there is extensive evidence that females are capable of evaluating and comparing male courtships, and accepting or rejecting males as mates. These relatively simple responses minimize random sexual encounters involving subpar conspecific males and heterospecific males, and over generations can potentially select novel physical and behavioral traits. Despite its evolutionary and behavioral significance, little is still known about the genes involved in mating choice and how choices for novel males and females arise during evolution. Drosophila simulans and Drosophila sechellia are two recently diverged species of Drosophila in which females have a preference for conspecific males. Here we analyzed a total of 1748 F2 hybrid females between these two species and found a small number of dominant genes controlling the preference for D. simulans males. We also mapped two redundant X-linked loci of mating choice, Macho-XA and Macho-XB, and show that neither one is required for female attractiveness. Together, our results reveal part of the genetic architecture that allows D. simulans females to recognize, mate, and successfully generate progenies with D. simulans males.


Spatial olfactory learning facilitates long-term depression in the hippocampus.
Recently, it has emerged that visual spatial exploration facilitates synaptic plasticity at different synapses within the trisynaptic network. Particularly striking is the finding that visuospatial contexts facilitate hippocampal long-term depression (LTD), raising the possibility that this form of plasticity may be important for memory formation. It is not known whether other sensory modalities elicit similar permissive effects on LTD. Here, we explored if spatial olfactory learning facilitates LTD in the hippocampus region of freely behaving rats. Patterned afferent stimulation of the Schaffer collaterals elicited short-term depression (STD) (<1 h) of evoked responses in the Stratum radiatum of the CA1 region. Coupling of this protocol with novel exploration of a spatial constellation of olfactory cues facilitated short-term depression into LTD that lasted for over 24 h. Facilitation of LTD did not occur when animals were re-exposed 1 week later to the same odors in the same spatial constellation. Evaluation of learning behavior revealed that 1 week after the 1st odor exposure, the animals remembered the odors and their relative positions. These data support that the hippocampus can use nonvisuospatial resources, and specifically can use spatial olfactory information, to facilitate LTD and to generate spatial representations. The data also support that a tight relationship exists between the processing of spatial contextual information and the expression of LTD in the hippocampus.


Impact of intensive upper limb rehabilitation on quality of life: a randomized trial in children with unilateral cerebral palsy.
The aim of this study was to determine whether constraint-induced movement therapy is more effective than bimanual training in improving the quality of life of children with unilateral cerebral palsy (CP). Sixty-three children (mean age 10y 2mo [SD 2y 6mo]; 33 males, 30 females) with CP of the spastic motor type (n=59) or with spasticity and dystonia (n=4) were randomly allocated to two groups. The children were assessed as Manual Ability Classification System level I (n=16), II (n=46), or III (n=1). Each group received 6 hours of daily intervention (either constraint-induced movement therapy [CIMT] or bimanual training [BIM]) for 10 days over a 2-week period (total intervention time 60h). Children aged 9 years and older completed the Cerebral Palsy Quality of Life Questionnaire for Children (CPQOL-Child) and those aged 8 years and older completed the KIDSCREEN-52. All parents completed proxy versions of each measure. Assessments were made at baseline and at 3, 26, and 52 weeks after the end of the intervention. Thirty-five children completed the CPQOL-Child and 41 completed the KIDSCREEN-52. No changes in social or emotional well-being were reported by children in either group. Children and parents from both groups reported a significant improvement in their or their child's feelings about functioning as well as participation and physical health on the CPQOL-Child. The parents of children receiving CIMT reported positive and sustained changes in their child's social well-being (CPQOL-Child). The CIMT group showed significant improvements in physical well-being, psychological well-being, and moods and emotions (KIDSCREEN-52) at 3 weeks post intervention, which were maintained over the study period. Intensive goal-directed upper limb training programmes using either CIMT or BIM achieved domain-specific changes in quality of life relating to feelings about functioning and participation and physical health. A condition-specific quality of life compared with a generic measure may be better able to detect changes in quality of life in children with unilateral CP.


Beyond-use dating of lidocaine alone and in two "magic mouthwash" preparations.
Beyond-use dating (BUD) of lidocaine alone and in two "magic mouthwash" preparations stored in amber oral syringes at room temperature was determined. Two formulations of mouthwash containing oral topical lidocaine 2% (viscous), diphenhydramine 2.5 mg/mL, and aluminum hydroxide-magnesium hydroxide-simethicone were prepared in 1:1:1 and 1:2.5:2.5 ratios, divided into 3-mL samples, and stored in unit-dose oral amber syringes. Unit-dose single-product lidocaine samples were also prepared to serve as controls and stored in oral amber syringes. The lidocaine concentrations in these samples were measured periodically for 90 days. A stability-indicating high-performance liquid chromatographic method was developed and validated for system suitability, accuracy, repeatability, intermediate precision, specificity, linearity, and robustness. Based on the calculated percentages versus the initial concentration and the results from an analysis of variance comparing the two formulations, a BUD of 21 days is deemed appropriate for both magic mouthwash formulations. Based on the stability data, published safety concerns, and lack of efficacy in combination, packaging and dispensing lidocaine separately from other ingredients are recommended when administering magic mouthwash mixtures. Utilizing a 90-day BUD, lidocaine can be packaged separately from other magic mouthwash ingredients in individual dosage units and applied to the oral cavity using the swish-and-spit method. The delivery of the diphenhydramine and aluminum hydroxide-magnesium hydroxide-simethicone could be separated, allowing for a swish-and-swallow method of administration. A BUD of 21 days is recommended for lidocaine prepared with diphenhydramine and aluminum hydroxide-magnesium hydroxide-simethicone in ratios of 1:1:1 and 1:2.5:2.5 and stored at room temperature in amber oral plastic syringes.


Temporomandibular joint pain as a manifestation of temporal arteritis.
We have described a patient whose temporal arteritis had several unusual manifestations. Except for a number of systemic complaints of longer duration, her predominant symptom was that of temporomandibular joint pain. Subsequent to the development of acute visual loss, her fundi remained normal, suggesting a posterior neuropathy. A high index of suspicion and meticulous history and physical examination should lead to early identification of temporal arteritis so that treatment can be instituted before permanent visual loss occurs.


Cloning and characterization of full-length mouse thymidine kinase 2: the N-terminal sequence directs import of the precursor protein into mitochondria.
The subcellular localization of mitochondrial thymidine kinase (TK2) has been questioned, since no mitochondrial targeting sequences have been found in cloned human TK2 cDNAs. Here we report the cloning of mouse TK2 cDNA from a mouse full-length enriched cDNA library. The mouse TK2 cDNA codes for a protein of 270 amino acids, with a 40-amino-acid presumed N-terminal mitochondrial targeting signal. In vitro translation and translocation experiments with purified rat mitochondria confirmed that the N-terminal sequence directed import of the precursor TK2 into the mitochondrial matrix. A single 2.4 kb mRNA transcript was detected in most tissues examined, except in liver, where an additional shorter (1.0 kb) transcript was also observed. There was no correlation between the tissue distribution of TK2 activity and the expression of TK2 mRNA. Full-length mouse TK2 protein and two N-terminally truncated forms, one of which corresponds to the mitochondrial form of TK2 and a shorter form corresponding to the previously characterized recombinant human TK2, were expressed in Escherichia coli and affinity purified. All three forms of TK2 phosphorylated thymidine, deoxycytidine and 2'-deoxyuridine, but with different kinetic efficiencies. A number of cytostatic pyrimidine nucleoside analogues were also tested and shown to be good substrates for the various forms of TK2. The active form of full-length mouse TK2 was a dimer, as judged by Superdex 200 chromatography. These results enhance our understanding of the structure and function of TK2, and may help to explain the mitochondrial disorder, mitochondrial neurogastrointestinal encephalomyopathy.


Semi-automatic scene generation using the Digital Anatomist Foundational Model.
A recent survey shows that a major impediment to more widespread use of computers in anatomy education is the inability to directly manipulate 3-D models, and to relate these to corresponding textual information. In the University of Washington Digital Anatomist Project we have developed a prototype Web-based scene generation program that combines the symbolic Foundational Model of Anatomy with 3-D models. A Web user can browse the Foundational Model (FM), then click to request that a 3-D scene be created of an object and its parts or branches. The scene is rendered by a graphics server, and a snapshot is sent to the Web client. The user can then manipulate the scene, adding new structures, deleting structures, rotating the scene, zooming, and saving the scene as a VRML file. Applications such as this, when fully realized with fast rendering and more anatomical content, have the potential to significantly change the way computers are used in anatomy education.


Implementation of a data accuracy plan to improve data extraction yield in the Axon Registry®.
To conduct a data quality improvement project to improve the quality measure data mapping and to measure key phrase logic in the Axon Registry.® METHODS: Prior validation analysis of the Axon Registry identified 2 main areas for remediation: methodology for mapping data from electronic health record (EHR) into the registry clinical data record (CDR) and key phrase logic for each measure. Practice groups participating in Axon Registry and 6 Axon Registry quality measures were selected for intervention. Mapping of measure elements and measure performances for each of the selected measures and practices were reviewed before intervention. The Data Accuracy Plan (DAP) was performed, and documentation data and visit data counts and data yield after intervention were calculated and analyzed. Documentation data and visit data counts and data yield increased for all 6 quality measures and all practices in the DAP. Increase in documentation data count ranged from 815 to 15,782 occurrences, while visit data count increase ranged from 519 to 16,383 visits. Average data yield range was 7.22% to 33.46% before intervention and increased to a range from 15.34% to 74.40% after intervention. There was substantial improvement in the accuracy of data extraction for quality measure elements after intervention to improve methodology for mapping EHR data into CDR and key phrase logic. Implementation of changes and continued review of data mapping and data dictionary are important to ensure accurate measure performance and to improve reliability and validity of Axon Registry data.