Local and Topical Anesthetics for Pediatric Patients in the Emergency Department.

Painful diagnostic and therapeutic procedures are common in the emergency department. Adequately treating pain, including the pain of procedures is an essential component of the practice of emergency medicine. Pain management is also part of the core competency for emergency medicine residencies and pediatric emergency medicine fellowships. There are many benefits to providing local and/or topical anesthesia before performing a medical procedure, including better patient and family satisfaction and increased procedural success rates. Local and topical anesthetics when used appropriately, generally, have few, if any, systemic side effects, such as hypotension or respiratory depression, which is an advantage over procedural sedation. Use of local and topical anesthetics can do much toward alleviating the pain and anxiety of pediatric patients undergoing procedures in the emergency department.

[Neuropsychology of psychoeducation in schizophrenia: results of the Munich COGPIP study]. / Neuropsychologie der Psychoedukation bei Schizophrenie: Ergebnisse der Münchner COGPIP-Studie.

BACKGROUND: The aim of the study was to examine whether the efficacy of psychoeducation in patients with schizophrenia is dependent on their cognitive performance and if a preceding cognitive training can enhance the therapeutic effects of psychoeducation. PATIENTS AND METHODS: A total of 116 inpatients were randomly assigned to either a standardized cognitive training (COGPACK) or to routine occupational therapy, followed by a psychoeducational group program of 8 sessions within 4 weeks for all study patients. The effects of cognitive training and psychoeducation were assessed directly afterwards and in a follow-up after 9 months. RESULTS: The patient knowledge and compliance improved. Neurocognition and especially memory acquisition significantly predicted illness knowledge after psychoeducation, whereas psychopathology did not. No differential effects of the COGPACK training were found. After 9 months 75% of the patients showed a very good compliance and the readmission rate was 18%. The results were comparable under both study conditions. CONCLUSION: Besides baseline illness knowledge neurocognition was the only significant predictor for illness knowledge after psychoeducation. Patients with cognitive deficits can profit from psychoeducation in the long run as well. In future it should be examined whether a modified cognitive training program could achieve a faster improvement of the illness knowledge.

Neurocognitive prediction of illness knowledge after psychoeducation in schizophrenia: results from the Munich COGPIP study.

BACKGROUND: Many patients with schizophrenia exhibit neurocognitive impairments, namely, in attentional, mnestic and executive functions. While these deficits limit psychosocial rehabilitation, their effect on psychoeducation is unknown. Within the framework of the longitudinal Munich Cognitive Determinants of Psychoeducation and Information in Schizophrenic Psychoses (COGPIP) study, we examined: (a) whether illness knowledge after psychoeducation could be predicted more precisely from the neurocognitive than from the psychopathological status of the patients; (b) which neurocognitive domains are best predictors. METHOD: A total of 116 in-patients with schizophrenic or schizoaffective disorders were randomized to a neurocognitive training or control condition (2 weeks) followed by a manualized psychoeducational group programme (4 weeks) and then observed over a 9-month follow-up. Repeated measurements included - among others - the Positive and Negative Syndrome Scale and a comprehensive neuropsychological test battery from which normative T scores were used to calculate one global and five domain-specific neurocognitive composite scores. Illness knowledge was measured by a questionnaire (WFB-52) tailored to the psychoeducational programme. RESULTS: Multiple linear regression analyses showed that, apart from baseline illness knowledge, neurocognition significantly predicted knowledge outcome as well as knowledge gain (measured by reliable change indices) after psychoeducation. This was not true for psychopathology. Among the domain-specific neurocognitive composite scores, only memory acquisition was a significant predictor of knowledge outcome and gain. CONCLUSIONS: Neurocognition, not psychopathology, is a significant predictor of illness knowledge after psychoeducation in schizophrenia. This finding should guide efforts to tailor psychoeducational interventions more closely to the patient's needs and resources.

Reconstitution of T cell receptor signaling in ZAP-70-deficient cells by retroviral transduction of the ZAP-70 gene.

A variant of severe combined immunodeficiency syndrome (SCID) with a selective inability to produce CD8 single positive T cells and a signal transduction defect in peripheral CD4+ cells has recently been shown to be the result of mutations in the ZAP-70 gene. T cell receptor (TCR) signaling requires the association of the ZAP-70 protein tyrosine kinase with the TCR complex. Human T cell leukemia virus type I-transformed CD4+ T cell lines were established from ZAP-70-deficient patients and normal controls. ZAP-70 was expressed and appropriately phosphorylated in normal T cell lines after TCR engagement, but was not detected in T cell lines from ZAP-70-deficient patients. To determine whether signaling could be reconstituted, wild-type ZAP-70 was introduced into deficient cells with a ZAP-70 retroviral vector. High titer producer clones expressing ZAP-70 were generated in the Gibbon ape leukemia virus packaging line PG13. After transduction, ZAP-70 was detected at levels equivalent to those observed in normal cells, and was appropriately phosphorylated on tyrosine after receptor engagement. The kinase activity of ZAP-70 in the reconstituted cells was also appropriately upregulated by receptor aggregation. Moreover, normal and transduced cells, but not ZAP-70-deficient cells, were able to mobilize calcium after receptor ligation, indicating that proximal TCR signaling was reconstituted. These results indicate that this form of SCID may be corrected by gene therapy.

The mechanism of the water expulsion vesicle of the ciliate Tetrahymena pyriformis.

Analysis of high-speed (150 frames/sec) cinematographs of the filling and expulsion of the water expulsion vesicle of Tetrahymena pyriformis shows that the vesicle fills as water is pumped into it by contractions of at least four ampullary sacs which are continuous with the endoplasmic reticulum. When filled, the vesicle is pressed against its two excretory pores by cyclotic movements of the cytoplasm. This pressure closes the apertures of the ampullae, preventing backflow from the vesicle into them, and also spreads the pellicle of and at the pore, thereby stretching and rupturing the pore-sealing membrane. The vesicle is then invaginated by the cytoplasmic pressure, driving fluid out of the pore. The pore-sealing membrane then reforms, apparently by constriction, and the vesicle is again filled. Electron micrographs show that crisscrossed pore-microtubules extend from the pore to the openings of the ampullae, anchoring the vesicle in place. Each pore is surrounded by a stack of at least 11 ring-microtubules, to which the anchoring pore-microtubules are attached. The pore-microtubules appear to exert tension which assists in spreading the pore, aiding cyclotic pressures in rupturing the pore-sealing membrane. A possible mechanism for the cyclotic pressure and ampullary contraction is proposed.

The mechanism of the nephridial apparatus of Paramecium multimicronucleatum. II. The filling of the vesicle by action of the ampullae.

Our recent analysis of the nephridial apparatus of Paramecium multimicronucleatum by high-speed cinematography (300 fps at X 250) indicates that before the water expulsion vesicle ("contractile vacuole") is completely voided of fluid during expulsion, the ampullae surrounding and confluent with the vesicle swell with fluid entering from their respective nephridial tubules. Once the membranes of the excretory pore at the base of the excretory canal (leading from the vesicle proper to the outside) have constricted and resealed the excretory pore, the up till then constricted injection tubules of the ampullae which conduct fluid to the vesicle open as waves of contraction along the coacervate gel around the ampulla and proceed along each ampulla from distal to proximal end. The coacervate gel around any one ampulla does not necessarily contract in phase with that of any other ampulla. Each ampulla acts independently. The fluid from the ampullae is thus pumped sequentially, but not in predetermined order, into the water expulsion vesicle, refilling and distending it. Our previous studies (Organ et al., 1968a) suggest that an actomyosinoid ATP-using mechanism may be functional in the ampullary contractions.

The mechanism of the nephridial apparatus of Paramecium multimicronucleatum. I. Expulsion of water from the vesicle.

Recent analysis of the mechanism of the nephridial apparatus of Paramecium multimicronucleatum by high-speed cinematography (300 fps at x 250) confirms the observations by electron microscopy (Schneider, 1960) that once the pore is opened, the vesicle is invaginated by adjacent cytoplasm and is emptied by collapsing under pressure from that cytoplasm, aided perhaps by pressure of the fibrils which anchor the ampullae to the excretory canal. There is no indication of active contraction of the vesicle or its membrane. There is no permanent pore to the vesicle. The vesicle is closed by a sealing of the ruptured membrane where it is in contact with the pellicular excretory canal. At onset of expulsion of vesicular fluid the membrane across the basal opening of the excretory canal is ripped along one semicircular portion of the excretory pore and is driven up against the opposite wall as a flap while the water rushes out. A constriction of the vesicular and cell membranes at the base of the excretory canal reseals the opening.

Pediatric major trauma: an approach to evaluation and management.

Trauma is the leading cause of death in children nationwide. Proper management of the pediatric trauma patient involves many of the components contained within standard trauma protocols. By paying strict attention to the anatomical and physiological differences in the pediatric population, clinicians will be assured the best possible outcomes. This article outlines the fundamentals of proper management of pediatric trauma patients.

How many photons are needed to reconstruct random objects in coherent X-ray diffractive imaging?

This paper presents an investigation of the reconstructibility of coherent X-ray diffractive imaging diffraction patterns for a class of binary random `bitmap' objects. Combining analytical results and numerical simulations, the critical fluence per bitmap pixel is determined, for arbitrary contrast values (absorption level and phase shift), both for the optical near- and far-field. This work extends previous investigations based on information theory, enabling a comparison of the amount of information carried by single photons in different diffraction regimes. The experimental results show an order-of-magnitude agreement.

The SH2-containing adapter protein GRB10 interacts with BCR-ABL.

Bcr-Abl is an oncogenic tyrosine kinase expressed in tumor cells of CML and a subset of ALL which in its unregulated and activated state is thought to cause cell transformation and leukemia. Bcr-Abl contains several autophosphorylation sites which serve as potential docking sites for SH2-containing signaling molecules. Mutational analysis has indicated that these autophosphorylation sites play a critical role in the transforming capability of Bcr-Abl. It has been shown that the SH2-containing adapter protein Grb2 binds to the autophosphorylation site Tyr(p)177 whereby it couples Bcr-Abl to the Ras pathway. The biological consequences of this interaction, however, are presently unclear. A Tyr177-mutated Bcr-Abl which lacks the ability to interact with the Grb2-SH2 domain still transforms myeloid cells and generates tumors in nude mice. We performed a yeast two-hybrid screen to identify signaling proteins which bind to distinct Bcr-Abl autophosphorylation sites. Autophosphorylation of Bcr-Abl in yeast was accomplished by using the DNA binding protein LexA which permits dimerization and crossphosphorylation of the fused bait. Using a LexA-Bcr-Abl full length fusion protein as bait, we identified several SH2-containing proteins. Among them we confirmed molecules already shown by others to interact with Bcr-Abl, in vivo, including Grb2, PI-3-kinase and Crk indicating that dimerization in yeast leads to autophosphorylation of tyrosine residues crucial for Bcr-Abl signaling in vivo. More importantly, we identified the SH2-containing protein Grb10 as a new binding partner for Bcr-Abl. This binding occurs in a phosphotyrosine-dependent manner at Bcr sites of Bcr-Abl. Both Abl and Bcr alone, as well as a kinase-defective Bcr-Abl, failed to interact with Grb10 in yeast. Mutational analysis uncovered a new SH2 binding site in Bcr-Abl located between Bcr aa242-446, which is different from the Grb2 binding site. Binding could be demonstrated in vitro and also in vivo as shown by co-immunoprecipitation analysis in CML cells. Using a temperature sensitive Bcr-Abl stably overexpressed in hematopoetic cells, we demonstrated that complex formation of Grb10 with Bcr-Abl was kinase activation-dependent in vivo. Notably, a Bcr-Abl mutant protein (Bcr/1-242-Abl) which lacks the ability to interact with Grb10 partially alleviated IL-3 dependence of Ba/F3 cells, indicating that the Grb10/Bcr-Abl interaction is important for Bcr-Abl-induced IL-3 independence of Ba/F3 cells. In addition, the Bcr/1-242-Abl mutant has a reduced capacity to induce focus formation in fibroblasts.

Large scale expression, purification and 2D crystallization of recombinant plant plasma membrane H+-ATPase.

P-type ATPases convert chemical energy into electrochemical gradients that are used to energize secondary active transport. Analysis of the structure and function of P-type ATPases has been limited by the lack of active recombinant ATPases in quantities suitable for crystallographic studies aiming at solving their three-dimensional structure. We have expressed Arabidopsis thaliana plasma membrane H+-ATPase isoform AHA2, equipped with a His(6)-tag, in the yeast Saccharomyces cerevisiae. The H+-ATPase could be purified both in the presence and in the absence of regulatory 14-3-3 protein depending on the presence of the diterpene fusicoccin which specifically induces formation of the H+-ATPase/14-3-3 protein complex. Amino acid analysis of the purified complex suggested a stoichiometry of two 14-3-3 proteins per H+-ATPase polypeptide. The purified H(+)-ATPase readily formed two-dimensional crystals following reconstitution into lipid vesicles. Electron cryo-microscopy of the crystals yielded a projection map at approximately 8 A resolution, the p22(1)2(1) symmetry of which suggests a dimeric protein complex. Three distinct regions of density of approximately equal size are apparent and may reflect different domains in individual molecules of AHA2.

Two-dimensional crystallization of a membrane protein on a detergent-resistant lipid monolayer.

Two-dimensional crystals of a membrane protein, the proton ATPase from plant plasma membranes, have been obtained by a new strategy based on the use of functionalized, fluorinated lipids spread at the air-water interface. Monolayers of the fluorinated lipids are stable even in the presence of high concentrations of various detergents as was established by ellipsometry measurements. A nickel functionalized fluorinated lipid was spread into a monolayer at the air-water interface. The overexpressed His-tagged ATPase solubilized by detergents was added to the subphase. 2D crystals of the membrane protein, embedded in a lipid bilayer, formed as the detergent was removed by adsorption. Electron microscopy indicated that the 2D crystals were single layers with dimensions of 10 microm or more. Image processing yielded a projection map at 9 A resolution, showing three well-separated domains of the membrane-embedded proton ATPase.

Cell cycle progression of chronic lymphocytic leukemia cells is controlled by cyclin D2, cyclin D3, cyclin-dependent kinase (cdk) 4 and the cdk inhibitor p27.

B-CLL cells are arrested in G0/early G1 phase of the cell cycle and are characterized by a marked hyporesponsiveness towards a variety of polyclonal B cell activators. We have previously demonstrated that costimulation with CpG-ODN and IL-2 can overcome this proliferative defect. Cyclin D3 is the principal D-type cyclin which mediates G1 progression in normal B cells, but in B-CLL cells both cyclin D2 and cyclin D3, were strongly upregulated upon stimulation. Both cyclins were associated with cdk4 but not with cdk6, which is the catalytic partner of D-type cyclins in normal B cells. Moreover, immune complexes consisting of cyclin D2 and cdk4 or cyclin D3 and cdk4 were both functional and phosphorylated the RB protein in vitro. The cell cycle inhibitor p27 plays a pivotal role in cell cycle progression of B lymphocytes and has been shown to be overexpressed in B-CLL cells. P27 was rapidly downregulated in B-CLL cells even when stimulated with a non-CpG-ODN or IL-2 alone, while only moderate regulation could be observed in normal B cells. Taken together, our findings demonstrate that regulation of early cell cycle progression differs between B-CLL cells and normal B cells. These findings do not only contribute to the understanding of B-CLL pathophysiology, but might ultimately lead to the identification of new therapeutic targets.

Tau and Abeta42 protein in CSF of patients with frontotemporal degeneration.

BACKGROUND: CSF concentrations of tau and beta-amyloid protein-42 (Abeta42) have been extensively studied in AD. Few data are available concerning CSF levels of both proteins in patients with frontotemporal degeneration (FTD). METHODS: The authors investigated CSF tau and Abeta42 concentrations in 34 patients with FTD, 74 patients with AD, and 40 cognitively healthy control subjects. CSF levels of tau and Abeta42 were measured by ELISA. With use of receiver operating characteristic-derived cutoff points and linear discrimination lines, the diagnostic sensitivity and specificity of both markers were determined. RESULTS: CSF tau concentrations were significantly higher in FTD than in control subjects but were significantly lower than in AD. CSF Abeta42 levels were significantly lower in FTD than in control subjects but were significantly higher than in AD. In subjects with FTD, neither tau nor Abeta42 levels correlated with the severity of dementia. The best discrimination between the diagnostic groups was obtained by simultaneous measurement of tau and Abeta42, yielding a sensitivity of 90% at a specificity of 77% (FTD vs controls) and a sensitivity of 85% at a specificity of 85% (FTD vs AD). CONCLUSIONS: In FTD, CSF levels of tau are elevated and Abeta42 levels are decreased. With use of these markers, subjects with FTD can be distinguished from control subjects and from patients with AD with reasonable accuracy.

Repetitive peripheral magnetic stimulation alleviates tactile extinction.

Despite its frequency in right brain damaged patients crucial mechanisms of tactile extinction are still obscure and treatments are unavailable. Recent PET observations suggest a hypometabolism in the primary and secondary somatosensory cortex of the lesioned hemisphere in patients with tactile extinction. Functional and morphological investigations have shown that the sensorimotor cortex has a remarkable capability of reorganization when the sensory inflow is changed. Repetitive peripheral magnetic stimulation (RPMS) applied in patients suffering from central paresis alleviates sensorimotor as well as cognitive deficits by the induction of proprioceptive inflow, thereby activating plasticity in the CNS. Based on the observation of reduced metabolic activity in patients suffering from tactile extinction we applied RPMS to explore the effects of peripheral sensory stimulation on tactile extinction. Fourteen right-hemisphere lesioned patients with tactile extinction were randomly allocated to an experimental and a control group. The experimental group received one single RPMS treatment of the left forearm as well as a condition of attentional cueing known to improve visual extinction. The control group, with comparable tactile extinction scores, neither received RPMS nor verbal cueing, but was tested twice to evaluate possible learning or test repetition effects. In the experimental group RPMS led to a significant reduction of left-sided extinctions in the recognition of different tactual surfaces, but had no effect on ipsilesional errors. In contrast, attentional cueing had no significant effect on left-sided extinction errors but unexpectedly increased right-hand extinction errors slightly but significantly. The control group showed stable extinction scores of the left- and right-hand stimulus across two measurements, thus ruling out learning or test repetition effects. These results show that sensory inflow is an important modulatory factor in tactile extinction. Furthermore, multiple RPMS may prove a promising way for the rehabilitation of patients with this disorder.

Alexithymia is no risk factor for exacerbation in spasmodic torticollis patients.

OBJECTIVE: To assess whether alexithymia is a risk factor for exacerbation in spasmodic torticollis (ST). METHODS: ST patients (2 x 10) with high vs. low alexithymia scores (mean score on the 20-item Toronto Alexithymia Scale [TAS-20]=69.2 vs. 28.7) were compared on physiological, motor and subjective responses to a cognitive and an emotional laboratory stressor. Changes in sustained abnormal head/shoulder positions and maximum range of motion (ROM) of the cervical spine were kinematically quantified. Skin conductance level (SCL), nonspecific skin conductance fluctuations (NS.SCF), heart rate (HR) and skin temperature (T) were measured. RESULTS: High alexithymia had no effect on the abnormal head posture or movements, but high-alexithymic ST patients showed generally increased levels of autonomic arousal (more NS.SCF, higher SCL; analysis of variance [ANOVA]: P=.016 and P=.051, respectively) under all experimental conditions. When ST symptom severity (TSUI-score) was partialled out, these group differences were somewhat reduced (analysis of covariance [ANCOVA]: P=.052 and P=.143). CONCLUSIONS: High alexithymia did not lead to increased abnormal head movements to stressors, but may result in a subtle increase in tonic level of sympathetic activity.

Three new scalarane-based sesterterpenes from the tropical marine sponge strepsichordaia lendenfeldi1

From the dichloromethane extract of the tropical marine sponge Strepsichordaia lendenfeldi collected from the Great Barrier Reef, Australia, three new (1, 2, and 9) and seven known (3-8 and 10) scalarane-based sesterterpenes were isolated. All molecular structures were secured by spectroscopic methods, particularly 1D and 2D NMR, and accurate mass measurement.

Monitoring of task performance during functional magnetic resonance imaging of sensorimotor cortex at 1.5 T.

Functional magnetic resonance imaging (fMRI) has found widespread clinical interest. Difficulties in clinical use of the fMRI technique arise, considering the lack of knowledge about activation task performance. This accounts especially for sensorimotor activation studies, in which performance of the sensorimotor activation task is-if at all-usually rated visually by subjective or semiquantitative methods (i.e., defining categories of performance such as neurological soft signs scales). Recently, instrumental methods for the measurement and analysis of motor performance have been developed. Pronation/supination (hand rotation) movement was shown to be an easily measurable and promising motor task. We have adapted a mechanic device (pronation/supination device, PSD) to monitor motor performance during the fMRI experiment. In a feasibility study, an investigation of fMRI activation strength dependence of sensorimotor cortices and supplementary motor area upon task frequency (25, 50, and 75 cycles/min) was carried out on 10 right-handed healthy volunteers. Furthermore, the authors report the observation of stimulus-induced activation changes in the cerebellum during pronation/supination movement.

Rasmussen encephalitis: epilepsia partialis continua secondary to chronic encephalitis.

Rasmussen encephalitis is a disease consisting of chronic encephalitis with progressive neurologic deficits and focal intractable seizure activity. The etiology is unknown, but pathologic specimens revealed changes consistent with viral encephalitis. Even though neuro-imaging techniques, such as positron emission tomography and magnetic resonance imaging, offer the prospect of specific, presurgical diagnostic criteria, the initial diagnosis usually is made on a clinical basis. Treatment modalities, including a wide variety of antiepileptic drug therapies and surgical interventions, may result in significant physical and mental impairments. We summarize the clinical presentation, diagnostic considerations, and different treatment protocols in a patient with this rare and debilitating disorder.