Some structural studies by deuterium magnetic resonance

Single crystals of ND 4 DS0 4 and Sr(DCOO)2.2D 2 0 have been studied by deuteron magnetic resonance spectroscopy; the apparatus and techniques used are described in some detail. ND 4 DS0 4 has considerable interest because of its ferroelectric properties: spectra were recorded at 300 K and 230 K, within the paraelectric and ferroelectric phases respectively. Symmetry relationships were used in solving the structure, particularly for refining the orientation data. At room temperature, the two sets of bisu1phate deuterons were found to have (within xperimental error) the same quadrupole coupling constants and asymmetry parameters: h = 191.9 ± 3KHz, T1 = 0.094 ± 0.02 ~ h = 193.2 ± 3KHz, T1 = 0.070 ± 0.0l. The quadrupolar constants appear somewhat larger than expected from the known 0····0 distances. No evidence of disorder is apparent from the room temperature data. At 230 K, both groups of deuterons appear to have suffered small angular displacements, essentially planar and antisymmetric with relation to the room temperature positions. One type of deuteron undergoes movements of larger amplitude in a plane more favourably oriented with respect to the polar axis, [Cl], which suggests a definite contribution from these deuterons to the appearance of ferroelectricity. The Curie point has been determined independently by use of the radio frequency spectrometer; a value of 266 ± 2K was obtained, in reasonable agreement with the literature value from dielectric constant measurements. The ND4+ group is reorienting rapidly both at 230 K and 300 K. Little change seems to occur upon transition. Some distortion of the tetrahedral structure, a possible difference in the reorientation rate for the two kinds of ND4+ ions, and a slight difference in their respective environments could be the main causes for the fine structure observed in the ammonium resonance. The study of Sr(DCOO)2.2D 2 0 gave the following average values for the quadrupolar constants and asymmetry parameters at 300 K: 2 formate ion [DC(1)0(1)0(2)]- ~ = 167.6 + 3KHz 'T] = 0.043 ± 0.01, DB h = 170.3 ± 3KHz 'T] = 0.025 ± 0.01. formate ion [DC(2)0(3)0(4)]- DB= h 242.8 + 3KHz 'T] = O. 100 ± 0.005 2 ~ = 216.8 + 3KHz 'T] = O. 113 ± 0.005 Both formate ions give e.f.g. tensors very close to axial symmetry. For [DC(1)0(1)0(2)]-, the deuteron is approximately aligned with the C(1) 0(10(2) bisectrix in an essentially planar arrangement. For [DC(2)0(3)0(4)]- the deuteron shows a surprising deviation of 22̊ from the plane of the COO group, as determined from Osaki's X-ray data. A weak C-D---O bonding interaction is tentatively suggested to account for this anomaly. D 2 0(1) was easily localised in the surrounding structure as known from X-ray data; the electric field gradient constants indicate that this molecule is 'static' at room temperature. A-D-0-D angle of 107.8̊ is in good agreement with the value for H2O D 2 D0(2) gave rise to broad and rather weak resonances with marked fine structure: some poorly defined curves were analysed only approximately. The high quadrupolar constants (330 ± 30 KHz) and asymmetry parameters (0.2 ± 0.1) derived for one set of the curves suggest that this D 2 0 molecule is weakly hydrogen bonded (if at all), and may have a rather high degree of motional freedom. analysed only approximately. The high quadrupolar constants (330 ± 30 KHz) and asymmetry parameters (0.2 ± 0.1) derived for one set of the curves suggest that this D 2 0 molecule is weakly hydrogen bonded (if at all), and may have a rather high degree of motional freedom

Characterization of α-Glucosidases From Lutzomyia longipalpis Reveals Independent Hydrolysis Systems for Plant or Blood Sugars

Lutzomyia longipalpis is the main vector of Leishmania infantum and exploits different food sources during development. Adults have a diet rich in sugars, and females also feed on blood. The sugar diet is essential for maintaining longevity, infection, and Leishmaniasis transmission. Carbohydrases, including α-glucosidases, are the main enzymes involved in the digestion of sugars. In this context, we studied the modulation of α-glucosidase activities in different feeding conditions and compartments of Lutzomyia longipalpis females, in order to characterize in detail their roles in the physiology of this insect. All tissues showed activity against MUαGlu and sucrose, with highest activities in the midgut and crop. Activity was 1,000 times higher on sucrose than on MUαGlu. Basal activities were observed in non-fed insects; blood feeding induced activity in the midgut contents, and sugar feeding modulated activity in midgut tissues. α-glucosidase activity changed after female exposure to different sugar concentrations or moieties. α-glucosidases from different tissues showed different biochemical properties, with an optimum pH around 7.0–8.0 and KM between 0.37 and 4.7 mM, when MUαGlu was used as substrate. Using sucrose as substrate, the optimum pH was around 6.0, and KM ranges between 11 and 800 mM. Enzymes from the crop and midgut tissues showed inhibition in high substrate concentrations (sucrose), with KI ranging from 39 to 400 mM, which explains the high KM values found. Chromatographic profiles confirmed that different α-glucosidases are been produced in L. longipalpis in different physiological contexts, with the distinction of at least four α-glucosidases. The results suggest that some of these enzymes are involved in different metabolic processes, like digestion of plant sugars, digestion of blood glycoproteins or glycolipids, and mobilization of energetic storages during starvation

Expression pattern of glycoside hydrolase genes in Lutzomyia longipalpis reveals key enzymes involved in larval digestion

The sand fly Lutzomyia longipalpis is the most important vector of American Visceral Leishmaniasis. Adults are phytophagous (males and females) or blood feeders (females only), and larvae feed on solid detritus. Digestion in sand fly larvae has scarcely been studied, but some glycosidase activities putatively involved in microorganism digestion were already described. Nevertheless, the molecular nature of these enzymes, as the corresponding genes and transcripts, were not explored yet. Catabolism of microbial carbohydrates in insects generally involves β-1,3-glucanases, chitinases, and digestive lysozymes. In this work, the transcripts of digestive β-1,3-glucanase and chitinases were identified in the L. longipalpis larvae throughout analysis of sequences and expression patterns of glycoside hydrolases families 16, 18, and 22. The activity of one i-type lysozyme was also registered. Interestingly, this lysozyme seems to play a role in immunity, rather than digestion. This is the first attempt to identify the molecular nature of sand fly larval digestive enzymes

Development of Leishmania mexicana in Lutzomyia longipalpis in the absence of sugar feeding

The leishmaniases are caused by Leishmania parasites and transmitted through the bites of phlebotomine sand flies. During parasite development inside the vector's midgut, promastigotes move towards the stomodeal valve, a mechanism that is crucial for transmission. It has been reported that the sugar meal acquired by sand flies during feeding between bloodmeals is essential for the development and migration of parasites. We demonstrated that the distribution of Leishmania mexicana parasites was affected by the sugar meals obtained by the sand flies. Promastigote migration towards the cardia region seems to be only partially based on the stimuli provided by sugar molecules. In the absence of sugars, significant amounts of parasites developed in the hindgut. In addition, sugar meals were important for the survival of sand flies, especially during blood digestion, presumably supporting their energy requirements

A report on the piloting of a novel computer-based medical case simulation for teaching and formative assessment of diagnostic laboratory testing

Objectives: Insufficient attention has been given to how information from computer-based clinical case simulations is presented, collected, and scored. Research is needed on how best to design such simulations to acquire valid performance assessment data that can act as useful feedback for educational applications. This report describes a study of a new simulation format with design features aimed at improving both its formative assessment feedback and educational function. Methods: Case simulation software (LabCAPS) was developed to target a highly focused and well-defined measurement goal with a response format that allowed objective scoring. Data from an eight-case computer-based performance assessment administered in a pilot study to 13 second-year medical students was analyzed using classical test theory and generalizability analysis. In addition, a similar analysis was conducted on an administration in a less controlled setting, but to a much large sample (n=143), within a clinical course that utilized two random case subsets from a library of 18 cases. Results: Classical test theory case-level item analysis of the pilot assessment yielded an average case discrimination of 0.37, and all eight cases were positively discriminating (range=0.11–0.56). Classical test theory coefficient alpha and the decision study showed the eight-case performance assessment to have an observed reliability of σ=G=0.70. The decision study further demonstrated that a G=0.80 could be attained with approximately 3 h and 15 min of testing. The less-controlled educational application within a large medical class produced a somewhat lower reliability for eight cases (G=0.53). Students gave high ratings to the logic of the simulation interface, its educational value, and to the fidelity of the tasks. Conclusions: LabCAPS software shows the potential to provide formative assessment of medical students’ skill at diagnostic test ordering and to provide valid feedback to learners. The perceived fidelity of the performance tasks and the statistical reliability findings support the validity of using the automated scores for formative assessment and learning. LabCAPS cases appear well designed for use as a scored assignment, for stimulating discussions in small group educational settings, for self-assessment, and for independent learning. Extension of the more highly controlled pilot assessment study with a larger sample will be needed to confirm its reliability in other assessment applications

Receptor-Mediated Enhancement of Beta Adrenergic Drug Activity by Ascorbate In Vitro and In Vivo

RATIONALE: Previous in vitro research demonstrated that ascorbate enhances potency and duration of activity of agonists binding to alpha 1 adrenergic and histamine receptors. OBJECTIVES: Extending this work to beta 2 adrenergic systems in vitro and in vivo. METHODS: Ultraviolet spectroscopy was used to study ascorbate binding to adrenergic receptor preparations and peptides. Force transduction studies on acetylcholine-contracted trachealis preparations from pigs and guinea pigs measured the effect of ascorbate on relaxation due to submaximal doses of beta adrenergic agonists. The effect of inhaled albuterol with and without ascorbate was tested on horses with heaves and sheep with carbachol-induced bronchoconstriction. MEASUREMENTS: Binding constants for ascorbate binding to beta adrenergic receptor were derived from concentration-dependent spectral shifts. Dose- dependence curves were obtained for the relaxation of pre-contracted trachealis preparations due to beta agonists in the presence and absence of varied ascorbate. Tachyphylaxis and fade were also measured. Dose response curves were determined for the effect of albuterol plus-and-minus ascorbate on airway resistance in horses and sheep. MAIN RESULTS: Ascorbate binds to the beta 2 adrenergic receptor at physiological concentrations. The receptor recycles dehydroascorbate. Physiological and supra-physiological concentrations of ascorbate enhance submaximal epinephrine and isoproterenol relaxation of trachealis, producing a 3-10-fold increase in sensitivity, preventing tachyphylaxis, and reversing fade. In vivo, ascorbate improves albuterol's effect on heaves and produces a 10-fold enhancement of albuterol activity in "asthmatic" sheep. CONCLUSIONS: Ascorbate enhances beta-adrenergic activity via a novel receptor-mediated mechanism; increases potency and duration of beta adrenergic agonists effective in asthma and COPD; prevents tachyphylaxis; and reverses fade. These novel effects are probably caused by a novel mechanism involving phosphorylation of aminergic receptors and have clinical and drug-development applications

The Treatment In Morning versus Evening (TIME) study:Analysis of recruitment, follow-up and retention rates post-recruitment

Abstract Background The use of information technology (IT) is now the preferred method of capturing and storing clinical research data. The Treatment In Morning versus Evening (TIME) study predominantly uses electronic data capture and IT to compare morning dosing of hypertensive medication against evening dosing. Registration, consent, participant demographics and follow-up data are all captured via the study website. The aim of this article is to assess the success of the TIME methodology compared with similar studies. Methods To assess the TIME study, published literature on similar clinical trials was reviewed and compared against TIME recruitment, follow-up and email interaction data. Results The TIME website registered 31,695 individuals, 21,116 of whom were randomised. Recruitment cost per randomised participant varied by strategy: £17.40 by GP practice, £3.08 by UK Biobank and £58.82 for GoShare. Twelve-month follow-up retention rates were 96%. A total of 1089 participants have withdrawn from their assigned time of dosing, 2% of whom have declined follow-up by record linkage or further contact. When the TIME data are compared with similar study data, study recruitment is very successful. However, TIME suffers difficulties with participant follow-up and withdrawal rates similar to those of conventional studies. Conclusions The TIME study has been successful in recruitment. Follow-up, retention rates and withdrawal rates are all acceptable, but ongoing work is required to ensure participants remain engaged with the study. Various recruitment strategies are necessary, and all viable options should be encouraged to maintain participant engagement throughout the life of studies using IT

Online clinical reasoning assessment with the Script Concordance test: a feasibility study

BACKGROUND: The script concordance (SC) test is an assessment tool that measures capacity to solve ill-defined problems, that is, reasoning in context of uncertainty. This tool has been used up to now mainly in medicine. The purpose of this pilot study is to assess the feasibility of the test delivered on the Web to French urologists. METHODS: The principle of SC test construction and the development of the Web site are described. A secure Web site was created with two sequential modules: (a) The first one for the reference panel (n = 26) with two sub-tasks: to validate the content of the test and to elaborate the scoring system; (b) The second for candidates with different levels of experience in Urology: Board certified urologists, residents, medical students (5 or 6(th )year). Minimum expected number of participants is 150 for urologists, 100 for residents and 50 for medical students. Each candidate is provided with an individual access code to this Web site. He/she may complete the Script Concordance test several times during his/her curriculum. RESULTS: The Web site has been operational since April 2004. The reference panel validated the test in June of the same year during the annual seminar of the French Society of Urology. The Web site is available for the candidates since September 2004. In six months, 80% of the target figure for the urologists, 68% of the target figure for the residents and 20% of the target figure for the student passed the test online. During these six months, no technical problem was encountered. CONCLUSION: The feasibility of the web-based SC test is successful as two-thirds of the expected number of participants was included within six months. Psychometric properties (validity, reliability) of the test will be evaluated on a large scale (N = 300). If positive, educational impact of this assessment tool will be useful to help urologists during their curriculum for the acquisition of clinical reasoning skills, which is crucial for professional competence

Acute epiglottitis as the initial presentation of pediatric Systemic Lupus Erythematosus

We report a case of a 5-year old girl, who initially presented with acute epiglottitis, sepsis and multi-organ failure. She was subsequently diagnosed as having Systemic Lupus Erythematosus. To the best of our knowledge, this article describes the first case of Haemophilus influenzae type f epiglottitis as the initial presentation of SLE in childhood