[Magnetic resonance imaging of subarachnoid hemorrhage].
Computed tomography (CT) is the method of choice to detect acute subarachnoid hemorrhage (SAH). The sensitivity of CT has been reported to range from 85 to 100 %. Magnetic resonance imaging (MRI) using FLAIR sequences shows a comparable sensitivity in acute SAH and in some cases can even be superior to CT. However, additional proton density weighted (PDW) TSE sequences or 3D-FLAIR sequences should be used to avoid false-positive results caused by flow artifacts. Conventional T (1)- or T (2)-weighted SE sequences or susceptibility weighted T (2)* or EPI sequences used for the diagnosis of intracerebral hemorrhage do not detect SAH reliably enough. In subacute SAH, starting from day 5 after the suspected hemorrhage, the sensitivity of MRI is clearly superior to CT. Patients with suspected ischemic stroke scheduled for the initial evaluation by MRI no longer need additional CT studies to rule out subarachnoid hemorrhage as long as the MRI protocols are adequate and the radiologist has the necessary experience.


Brain white-matter lesions and psychosis.
In a prospective study of late-life onset psychosis, five of the first 27 patients studied had extensive white-matter lesions demonstrated by MRI and/or CT. None of 60 age-matched psychiatrically healthy controls demonstrated such lesions. All five patients had a mild dementia and a frontal behavioural syndrome. In addition, every patient performed poorly on neuropsychological tests of frontal function. Dysfunction of the frontal cortex associated with white-matter lesions appears to contribute to the clinical picture of some cases of late-life psychosis.


Molecular identification of livestock breeds: a tool for modern conservation biology.
Global livestock genetic diversity includes all of the species, breeds and strains of domestic animals, and their variations. Although a recent census indicated that there were 40 species and over 8000 breeds of domestic animals; for the purpose of conservation biology the diversity between and within breeds rather than species is regarded to be of crucial importance. This domestic animal genetic diversity has developed through three main evolutionary events, from speciation (about 3 million years ago) through domestication (about 12000 years ago) to specialised breeding (starting about 200 years ago). These events and their impacts on global animal genetic resources have been well documented in the literature. The key importance of global domestic animal resources in terms of economic, scientific and cultural heritage has also been addressed. In spite of their importance, there is a growing number of reports on the alarming erosion of domestic animal genetic resources. This erosion of is happening in spite of several global conservation initiatives designed to mitigate it. Herein we discuss these conservation interventions and highlight their strengths and weaknesses. However, pivotal to the success of these conservation initiatives is the reliability of the genetic assignment of individual members to a target breed. Finally, we discuss the prospect of using improved breed identification methodologies to develop a reliable breed-specific molecular identification tool that is easily applicable to populations of livestock breeds in various ecosystems. These identification tools, when developed, will not only facilitate the regular monitoring of threatened or endangered breed populations, but also enhance the development of more efficient and sustainable livestock production systems.


Thermal and thermomechanical properties of poly(butylene succinate) nanocomposites.
This article describes the thermal and thermomechanical properties of poly(butylene succinate) (PBS) and its nanocomposites. PBS nanocomposites with three different weight ratios of organically modified synthetic fluorine mica (OMSFM) have been prepared by melt-mixing in a batch mixer at 140 degrees C. The structure and morphology of the nanocomposites were characterized by X-ray diffraction (XRD) analyses and transmission electron microscopy (TEM) observations that reveal the homogeneous dispersion of the intercalated silicate layers into the PBS matrix. The thermal properties of pure PBS and the nanocomposite samples were studied by both conventional and temperature modulated differential scanning calorimetry (DSC) analyses, which show multiple melting behavior of the PBS matrix. The investigation of the thermomechanical properties was performed by dynamic mechanical analysis. Results reveal significant improvement in the storage modulus of neat PBS upon addition of OMSFM. The tensile modulus of neat PBS is also increased substantially with the addition of OMSFM, however, the strength at yield and elongation at break of neat PBS systematically decreases with the loading of OMSFM. The thermal stability of the nanocomposites compared to that of the pure polymer sample was examined under both pyrolytic and thermo-oxidative environments. It is shown that the thermal stability of PBS is increased moderately in the presence of 3 wt% of OMSFM, but there is no significant effect on further silicate loading in the oxidative environment. In the nitrogen environment, however, the thermal stability systematically decreases with increasing clay loading.


The lifespan trajectory of neural oscillatory activity in the motor system.
Numerous studies connect beta oscillations in the motor cortices to volitional movement, and beta is known to be aberrant in multiple movement disorders. However, the dynamic interplay between these beta oscillations, motor performance, and spontaneous beta power (e.g., during rest) in the motor cortices remains unknown. This study utilized magnetoencephalography (MEG) to investigate these three parameters and their lifespan trajectory in 57 healthy participants aged 9-75 years old. Movement-related beta activity was imaged using a beamforming approach, and voxel time series data were extracted from the peak voxels in the primary motor cortices. Our results indicated that spontaneous beta power during rest followed a quadratic lifespan trajectory, while movement-related beta oscillations linearly increased with age. Follow-on analyses showed that spontaneous beta power and the beta minima during movement, together, significantly predicted task performance above and beyond the effects of age. These data are the first to show lifespan trajectories among measures of beta activity in the motor cortices, and suggest that the healthy brain compensates for age-related increases in spontaneous beta activity by increasing the strength of beta oscillations within the motor cortices which, when successful, enables normal motor performance into later life.


Cardiovascular effects of mianserin and amitriptyline in healthy volunteers.
10 healthy volunteers were randomized in two groups and treated with mianserin and amitriptyline for 6 days. Heart rate, arterial blood pressure, and blood levels of the drugs were measured daily. ECG and left-ventricular echocardiogram as a parameter of contractility were registered before as well as after 3 and 6 days of treatment with both drugs. The most striking result of our investigation was that amitriptyline-treated subjects showed a loss of myocardial contractility (-17% of the initial value) after 6 days of treatment; mianserin was ineffective in this respect. After both mianserin and amitriptyline an increase of the mean arterial blood pressure was observed; an increased heart rate was seen only in the amitriptyline group. No alterations in the parameters calculated from the ECG registrations were found in either group. The inability of mianserin to decrease cardiac contractility in the therapeutic dose range may favor this tetracyclic antidepressant over the tricyclic compound amitriptyline, especially in patients with cardiac disease.


[Features of the angioarchitectonics of the papillary muscles and the chordae tendineae of the human heart].
In persons of mature, elderly and old age vascularization of the papillary muscles and tendinous chordae in the right and left cardiac ventricles has been studied depending on peculiarities of their structure and sources of their blood supply using injection, macro-microscopical, roentgenoangiographic methods, and silver nitrate impregnation. Certain data concerning the distribution zones in the papillary muscles of the coronary artery branches at various types of the cardiac blood supply are presented. Angioarchitectonics of the papillary muscles are described along their whole extention--in the basal area, in the middle part and at the apex. Architectonic peculiarities of the blood vessels in the muscular-tendinous part of the papillary muscles and in some tendinous chordae are described.


Care platforms: a basic building block for care delivery.
Without significant operational reform within the nation's health care delivery organizations, new financing models, payment systems, or structures are unlikely to realize their promise. Adapting insights from high-performing companies in other high-risk, high-cost, science- and technology-based industries, we propose the "care platform" as an organizing framework for internal operations in diversified provider organizations to increase the quality, reliability, and efficiency of care delivery. A care platform organizes "care production" around similar work, rather than organs or specialties; integrates standard and custom care processes; and surrounds them with specifically configured information and business systems. Such organizational designs imply new roles for physicians.


Neoparamoeba Page, 1987: light and electron microscopic observations on six strains of different origin.
Although amoebic gill disease (AGD) has emerged as one of the most severe health problems in the fish industry, proof of the identity of AGD agents from various localities is still missing. Six strains of amoebae designated until recently as Paramoeba species (the agents of AGD) were studied in cultures by light and electron microscopy. Although they were isolated from gills of different hosts (Dicentrarchus labrax and Scophthalmus maximus) and from distant localities, their morphology was identical. The strains differed from Paramoeba eilhardi, the type species of the genus, in that they lacked the boat-shaped microscales on the cell surface but could be safely identified as belonging to the genus Neoparamoeba Page, 1987. Transmission electron microscopy revealed the presence of a symbiotic organism, Perkinsiella amoebae Hollande, 1980, in all strains under study. The only difference among the strains examined was found in the size of trophozoites, which could be attributed to the different origins of the strains, but until more refined diagnostic methods are available, in addition to N. pemaquidensis, the closely related species N. aestuarina also has to be taken into consideration as the agent of AGD.


Cyclic AMP-induced slow inward current in depolarized neurons of Aplysia californica.
Cyclic nucleotides have been implicated in many long-lasting transmitter-induced effects on membrane conductance. One previously observed effect of cAMP on molluscan neurons is to induce a slow inward current, which has been further evaluated here in depolarized anterior and medial cells of the pleural ganglion of Aplysia californica in order to understand better its underlying ionic mechanisms and its sensitivity to a variety of pharmacological agents. This current, which appears to be the only cAMP-induced current seen in the anterior cells, was shown to invert at about +25 mV, that is, approximately 25-30 mV inferior to ENa. This reversal potential was lowered by about 15-16 mV when half of the extracellular Na was replaced by either mannitol or N-methyl-D-glucamine, whereas it was unaffected by changes in extracellular Cl, Ca, or Mg. The response persisted in seawater in which the Na had been totally replaced by K, and its reversal potential shifted towards more negative values. These data are consistent with the hypothesis that both Na and K ions permeate the channel, with a Na/K permeability ratio of approximately 2. Ca ions do not appear to permeate the channel, but they do have a marked inhibitory effect on the response amplitude, as do Mg ions when Ca is not present. Caffeine, intracellular acidification, and phosphodiesterase inhibitors enhance and prolong the response without changing its reversal potential. Previous studies have shown that both caffeine and intracellular acidification inhibit phosphodiesterase, and it is assumed that the common effect of these manipulations on the cAMP-induced inward current is mediated, at least partially, by the inhibition of that enzyme. In the medial cells of the pleural ganglion, this slow inward current is present, but is dominated in the depolarized cell by a cAMP-induced diminution in a Ca-activated K conductance (Kehoe, 1985b). This K conductance and, consequently, the noninverting, cAMP-induced inward current that reflects its diminution, were shown to disappear in Ca-free solutions, in the presence of isobutyl-1-methylxanthine (IBMX) or caffeine, and upon acidification of the cytoplasm. When this cAMP-sensitive K conductance is blocked, the presence of the inverting cAMP-induced cationic current is unmasked. The cAMP-induced cationic current is shown to have many properties in common with cyclic nucleotide-induced currents described in photoreceptors, olfactory receptor cilia, and cardiac myocytes, all of which have been shown to be outwardly rectifying cationic currents that are inhibited by divalent cations and do not involve the activation of a cAMP-dependent kinase.


Mood disorders in patients with epilepsy: epidemiology and management.
Patients with epilepsy are at high risk for depression because of an incompletely understood combination of factors that may be both psychosocial and neurological. Interictal depression in patients with epilepsy is an undertreated condition, in part because of concern regarding drug interactions and the risk of exacerbating seizures with antidepressant treatment. Bipolar disorder is not described as occurring with a higher than expected frequency in the population with epilepsy, but high rates of depression and suicide are well recognised, highlighting the need for more emphasis on antidepressive treatment in this group of at-risk patients. Neurological factors, including site and lateralisation of seizure focus, may be important for the development of depression, with left-sided seizure foci having a higher association with depressive symptoms. Forced normalisation may be a factor in the paradoxical onset of depression in patients with epilepsy whose seizures suddenly become well controlled by anti-seizure treatment. Lowering of folic acid levels by some antiepileptic drugs (AEDs) may also influence the expression of depression in patients with epilepsy. New AEDs continue to emerge as beneficial treatments themselves for mood disorders, with lamotrigine, gabapentin and, to a lesser extent, topiramate having clinical trials data to support their use in patients with bipolar disease. Similar positive data are available for vagal nerve stimulation. Mood effects of AEDs can be complicated, however, as many of these drugs (e.g. tiagabine) have also been reported to cause depression as an adverse effect. Electroconvulsive therapy in depressed patients with epilepsy requires special consideration. The selective serotonin reuptake inhibitors (SSRIs) and antidepressants that act at multiple receptors (e.g. nefazodone, venlafaxine) are the most appropriate treatments for depressed patients with epilepsy. Among these agents, citalopram has a low risk of interactions with AEDs. Bupropion, clomipramine and maprotiline are associated with a greater risk of seizures compared with other antidepressants and consequently should be used with caution in the treatment of depression in patients with epilepsy.


Parietal epithelial cells maintain the epithelial cell continuum forming Bowman's space in focal segmental glomerulosclerosis.
In the glomerulus, Bowman's space is formed by a continuum of glomerular epithelial cells. In focal segmental glomerulosclerosis (FSGS), glomeruli show segmental scarring, a result of activated parietal epithelial cells (PECs) invading the glomerular tuft. The segmental scars interrupt the epithelial continuum. However, non-sclerotic segments seem to be preserved even in glomeruli with advanced lesions. We studied the histology of the segmental pattern in Munich Wistar Frömter rats, a model for secondary FSGS. Our results showed that matrix layers lined with PECs cover the sclerotic lesions. These PECs formed contacts with podocytes of the uninvolved tuft segments, restoring the epithelial continuum. Formed Bowman's spaces were still connected to the tubular system. In biopsies of patients with secondary FSGS, we also detected matrix layers formed by PECs, separating the uninvolved from the sclerotic glomerular segments. PECs have a major role in the formation of glomerulosclerosis; we show here that in FSGS they also restore the glomerular epithelial cell continuum that surrounds Bowman's space. This process may be beneficial and indispensable for glomerular filtration in the uninvolved segments of sclerotic glomeruli.


Tumor Location Based Segmentation in Upper-Tract Urothelial Carcinoma Impacts on the Urothelial Recurrence-Free Survival: A Multi-Institutional Database Study.
The predictive impact of primary tumor location for patients with upper-tract urothelial carcinoma (UTUC) in the presence of concomitant urothelial bladder cancer, along with urothelial recurrence after the curative treatment is still contentious. We evaluated the association between precise tumor location and concomitant presence of urothelial bladder cancer and urothelial recurrence-free survival in patients with UTUC treated by radical nephroureterectomy with a bladder cuff. A total of 1,349 patients with localized UTUC (Ta-4N0M0) from a retrospective multi-institutional cohort were studied. We queried four UTUC databases. This retrospective clinical series was of patients with localized UTUC managed by nephroureter-ectomy with a bladder cuff, for whom data were from the Nishinihon Uro-Oncology Collaborative Group registries. Patients with a history of chemotherapy or radiotherapy were excluded from the study. Associations between the location of the tumor and subsequent outcome following nephroureterectomy were assessed using COX multivariate analysis. The location of the tumor was verified by pathological samples. Urothelial recurrence was defined as tumor relapse in any local urothelium, and coded apart from distant metastasis. The median follow-up was 34 months. A total of 887 patients had an evaluation of the tumor location in which 475 patients had pelvic tumors (53.6%), 96 had ureteral tumors in the U1 segment (10.8%), 87 in the U2 segment (9.8%), and 176 in the U3 segment (19.8%). There were 52 patients who had multifocal tumors (5.9%) as follows: 8 (0.9%) in the pelvis and ureter, 11 (1.2%) in U1 + U2, 1 (0.1%) in U1 + U3, 27 (3.0 %) in U2 + U3, and 6 (0.7%) in U1 + U2 + U3. In all, 145 (16.3%) had concomitant bladder tumors. Logistic regression analysis of gender, age, hydronephrosis, cytology, performance status, grade, lymphovascular invasion, pT, pN, and tumor focality showed that tumor location was associated with the presence of concomitant bladder cancer (p = 0.004, HR = 1.265). When the tumor location was stratified into 8 segments, including multifocal tumors, only the U3 segment remained as a predictor for the presence of concomitant bladder cancer (p = 0.002, HR = 2.872). Kaplan-Meier analysis for unifocal disease showed that lower ureter tumors (a combination of U2 and U3) had a worse prognosis for urothelial recurrence than pelvic tumors or upper ureteral tumors (U1) (p < 0.001 for lower ureteral tumors versus pelvic tumors, p = 0.322 for upper ureteral tumor versus pelvic tumor by log rank). Multivariate analysis showed that lower ureter remained as a prognostic factor for urothelial recurrence after adjusting for gender, age, hydronephrosis, urine cytology, lymphovascular invasion, pT, and pN (p < 0.001, HR = 1.469), and a similar tendency was found when the analysis was run for patients without concomitant bladder tumors (p = 0.003, HR = 1.446). Patients with lower ureteral tumors had a higher prevalence of deaths (HR = 2.227) compared to patients with upper ureter tumors. This multi-institutional study showed that the primary tumor locations were independently associated with the presence of concomitant bladder tumors and subsequent urothelial recurrence.


Cranial nerve XII: the hypoglossal nerve.
The hypoglossal nerve provides motor innervation to the tongue, and may be affected by pathology at multiple locations along its pathway. Knowledge of its anatomy and careful examination of the tongue are important in the diagnosis of lesions and determination of appropriate testing. The hypoglossal nerve anatomy and physiology, clinical disorders, and the evaluation of the hypoglossal nerve are reviewed.


Everolimus in kidney transplant recipients at high cardiovascular risk: a narrative review.
Kidney transplant recipients (KTRs) are at increased risk of cardiovascular (CV) morbidity and mortality, and side effects induced by immunosuppressive therapy may be a major contributor to this risk, together with traditional CV risk factors. Many strategies have been considered in order to reduce CV risk in KTRs, such as steroid and/or calcineurin inhibitor (CNI) minimization, but current data are inconclusive. The introduction of mammalian target of rapamycin (mTOR) inhibitors, the cornerstone of CNI minimization, in the immunosuppressive protocol may reduce both the incidence and severity of CNI-associated side effects; however, whether this strategy has an impact on CV risk after kidney transplantation needs to be evaluated. To this end, a panel of Italian experts in the field of transplantation was convened in a series of meetings to assess the current literature on the potential of the mTOR inhibitor everolimus as a cardioprotective agent. This narrative review summarizes the panel's round-table discussions and provides recommendations for CV risk management in KTRs.


Effect of CNT/PDMS Nanocomposites on the Dynamics of Pioneer Bacterial Communities in the Natural Biofilms of Seawater.
In this study, the antifouling (AF) performance of different carbon nanotubes (CNTs)-modified polydimethylsiloxane (PDMS) nanocomposites (PCs) was examined directly in the natural seawater, and further analyzed using the Multidimensional Scale Analyses (MDS) method. The early-adherent bacterial communities in the natural biofilms adhering to different PC surfaces were investigated using the single-stranded conformation polymorphism (SSCP) technique. The PCs demonstrated differences and reinforced AF properties in the field, and they were prone to clustering according to the discrepancies within different CNT fillers. Furthermore, most PC surfaces only demonstrated weak modulating effects on the biological colonization and successional process of the early bacterial communities in natural biofilms, indicating that the presence of the early colonized prokaryotic microbes would be one of the primary causes of colonization and deterioration of the PCs. C6 coating seems to be promising for marine AF applications, since it has a strong perturbation effect on pioneer prokaryotic colonization.


UDP is a competitive antagonist at the human P2Y14 receptor.
G protein-coupled P2Y receptors (P2Y-R) are activated by adenine and uracil nucleotides. The P2Y(14) receptor (P2Y(14)-R) is activated by at least four naturally occurring UDP sugars, with UDP-glucose (UDP-Glc) being the most potent agonist. With the goal of identifying a competitive antagonist for the P2Y(14)-R, UDP was examined for antagonist activity in COS-7 cells transiently expressing the human P2Y(14)-R and a chimeric Galpha protein that couples Gi-coupled receptors to stimulation of phosphoinositide hydrolysis. UDP antagonized the agonist action of UDP-Glc, and Schild analysis confirmed that the antagonism was competitive (pK(B) = 7.28). Uridine 5'-O-thiodiphosphate also antagonized the human P2Y(14)-R (hP2Y(14)-R) with an apparent affinity similar to that of UDP. In contrast, no antagonist activity was observed with ADP, CDP, or GDP, and other uracil analogs also failed to exhibit antagonist activity. The antagonist activity of UDP was not observed at other hP2Y-R. In contrast to its antagonist action at the hP2Y(14)-R, UDP was a potent agonist (EC(50) = 0.35 muM) at the rat P2Y(14)-R. These results identify the first competitive antagonist of the P2Y(14)-R and demonstrate pharmacological differences between receptor orthologs.


Oncogene addiction in non-small cell lung cancer: Focus on ROS1 inhibition.
Detection of molecular aberrations driving the biology and the clinical behavior of advanced non-small cell lung cancer (NSCLC) allows the adoption of specific therapeutic strategies dramatically impacting disease courses. Among these, ROS1 rearrangements are present in 1-2% of lung adenocarcinomas. Thanks to similarities between ALK and ROS1 oncogenes, lessons inferred from ALK can be applied to ROS1-positive NSCLC; nevertheless, disparities exist between diseases mastered by these two fusion genes. In the absence of more common genetic alterations detected in NSCLC (e.g. EGFR and KRAS mutations, ALK gene fusions), seeking for ROS1 rearrangements is crucial. Dedicated molecular diagnostics should be standardized, hopefully relying upon practical and efficient algorithms, comprehending immunohistochemistry and fluorescence in situ hybridisation. The major clinical impact exerted by crizotinib represents the main reason for which not even a sole ROS1-positive tumor should be undetected. The recent approval of the inhibitor by both American and European health agencies would hopefully boost the widespread testing for ROS1, eventually increasing the absolute number of positive cases, potential further source of information regarding molecular and clinical resistance. In vitro and clinical evidence have already been generated concerning crizotinib resistance and strategies to maintain patients under specific driver-inhibition are being successfully developed. Gathering data concerning diagnostics, preclinical evidence, clinical practice and ongoing studies, the present review depicts the current scenario of ROS1 inhibition in NSCLC.


Bioelectrical brain effects of one's own voice identification in pitch of voice auditory feedback.
Control of voice fundamental frequency (F0) relies in part on comparison of the intended F0 level and auditory feedback. This comparison impacts "sense of agency", or SoA, commonly defined as being the agent of one's own actions and plays a key role for self-awareness and social interactions. SoA is aberrant in several psychiatric disorders. Knowledge about brain activity reflecting SoA can be used in clinical practice for these disorders. It was shown that perception of voice feedback as one's own voice, reflecting the recognition of SoA, alters auditory sensory processing. Using a voice perturbation paradigm we contrasted vocal and bioelectrical brain responses to auditory stimuli that differed in magnitude: 100 and 400 cents. Results suggest the different magnitudes were perceived as a pitch error in self-vocalization (100 cents) or as a pitch shift generated externally (400 cents). Vocalizations and neural responses to changes in pitch of self-vocalization were defined as those made to small magnitude pitch-shifts (100 cents) and which did not show differential neural responses to upward versus downward changes in voice pitch auditory feedback. Vocal responses to large magnitude pitch shifts (400 cents) were smaller than those made to small pitch shifts, and neural responses differed according to upwards versus downward changes in pitch. Our results suggest that the presence of SoA for self-produced sounds may modify bioelectrical brain responses reflecting differences in auditory processing of the direction of a pitch shift. We suggest that this modification of bioelectrical response can be used as a biological index of SoA. Possible neuronal mechanisms of this modification of bioelectrical brain response are discussed.


Origins and evolutionary diversification of the nuclear receptor superfamily.
Nuclear receptors (NRs), which include those for steroid and thyroid hormones as well as retinoids, are encoded by a large gene superfamily that has evolved to regulate nearly every facet of metazoan life, from development to basic metabolism. This article reviews the conservation in structure and function of distinct receptors across different species and attempts to draw conclusions as to the evolution of this gene superfamily. Although sequences related to NRs can be found in plants and yeast, gene sequence analyses suggest that the NR ancestor(s) first appeared in the early metazoans and subsequently diversified into the six receptor sub-families, which were already recognisable at the time of the Arthropoda-Chordata split over 700 million years ago. At the time when a primitive NR emerged, the basic components of the transcription regulatory machinery, which are conserved from yeast to vertebrates, were already in place and the ancestral NR must have evolved with the ability to communicate with them. The first such NRs likely acted as monomers and in a ligand-independent fashion. As members of the NR superfamily acquired the ability to hetero- and homodimerise, and to bind and be regulated by ligands, the functional complexity of the NR superfamily increased. This exponentially increasing complexity subsequently provided a potential driving force for evolution of higher organisms by supplying a sophisticated regulatory gene network that could control complex physiological processes during development and in adult organisms.


Development of a novel bioinformatics tool for in silico validation of protein interactions.
Protein interactions are crucial in most biological processes. Several in silico methods have been recently developed to predict them. This paper describes a bioinformatics method that combines sequence similarity and structural information to support experimental studies on protein interactions. Given a target protein, the approach selects the most likely interactors among the candidates revealed by experimental techniques, but not yet in vivo validated. The sequence and the structural information of the in vivo confirmed proteins and complexes are exploited to evaluate the candidate interactors. Finally, a score is calculated to suggest the most likely interactors of the target protein. As an example, we searched for GRB2 interactors. We ranked a set of 46 candidate interactors by the presented method. These candidates were then reduced to 21, through a score threshold chosen by means of a cross-validation strategy. Among them, the isoform 1 of MAPK14 was in silico confirmed as a GRB2 interactor. Finally, given a set of already confirmed interactors of GRB2, the accuracy and the precision of the approach were 75% and 86%, respectively. In conclusion, the proposed method can be conveniently exploited to select the proteins to be experimentally investigated within a set of potential interactors.


Schiff-base deprotonation is mandatory for light-dependent rhodopsin phosphorylation.
The absorption of light by rhodopsin leads to the formation of an activated intermediate (R*) capable of catalysing the exchange of GTP for GDP in a retinal guanine-nucleotide-binding regulatory protein (transducin). The ability of R* to function as a catalyst is terminated by the rhodopsin kinase. The 10 nonactive-site lysine residues of rhodopsin can be reductively dimethylated to form permethylated rhodopsin (PMRh). This derivative is phosphorylated to the same extent as rhodopsin after photolysis. The monomethylation of the active-site lysine residue of PMRh yields active-site-methylated rhodopsin (AMRh). It had previously been shown, by using AMRh, that the formation of R* and its spectroscopic signature metarhodopsin II requires the photochemically induced deprotonation of the active-site Schiff base [Longstaff, Calhoon & Rando (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 4209-4213]. Here it is demonstrated that active-site Schiff-base deprotonation is also mandatory in the formation of the form of photolyzed rhodopsin that is susceptible to phosphorylation by rhodopsin kinase. In terms of the spectroscopically defined rhodopsin intermediates, this means that only metarhodopsin II and possibly metarhodopsin III are the actual substrates for rhodopsin kinase.


[A woman with severe abdominal pain and endometriosis in the past].
A 44-year-old woman came to the emergency department with severe pain in the right upper abdomen. Her medical history mentioned a low anterior resection 8 years ago because of severe endometriosis. The CT scan showed a ruptured right hemidiaphragm with herniation of the small intestine.


Molecular cytogenetics of the ribosomal (18S + 28S and 5S) DNA loci in primitive and advanced urodele amphibians.
In the present work we performed a cytogenetic analysis of the ribosomal (18S + 28S and 5S) loci in amphibian species belonging to the advanced family Salamandridae (genera Triturus, Salamandra, and Salamandrina) and in the primitive hynobiid Salamandrella keyserlingii (family Hynobiidae). In each analyzed karyotype the 5S rDNA sites appear to be stable, and definite in number, while an intraspecific variability both in number and chromosomal location of the 18S + 28S rDNA loci has been found in some Triturus species. In particular, an evolutionary trend toward a large intraspecific variability of the 18S + 28S rDNA loci has been found in the T. vulgaris species group. A structural analysis of the ribosomal repetition units demonstrates the occurrence of a length polymorphism within the 18S + 28S rDNA repeats in the examined species of the family Salamandridae; however, this polymorphism is rather limited, even in those Triturus species characterized by high intragenomic variability of the ribosomal sites. We show that in T. vulgaris meridionalis the variant repetition units actually segregate with individual chromosomes. This implies that they are not intermingled in the ribosomal clusters.


Two-ventricle repair for complex congenital heart defects palliated towards single-ventricle repair.
Complex congenital heart defects that present earlier in life are sometimes channelled towards single-ventricle repair, because of anatomical or logistic challenges involved in two-ventricle correction. Given the long-term functional and survival advantage, we have been consciously exploring the feasibility of a biventricular repair in these patients when they present later for Fontan completion. Since June 2009, 71 patients were referred for staged completion of the Fontan procedure. Following detailed evaluation that included three-dimensional echocardiography and magnetic resonance imaging, 10 patients (Group 1-median age 6 years) were identified and later underwent complex biventricular repair with takedown of Glenn shunt, while completion of extracardiac Fontan repair was done in 61 patients (Group 2-median age 7 years). Two-ventricle repair was accomplished in all the 10 Group 1 patients. One patient developed complete heart block requiring permanent pacemaker insertion. Late patch dehiscence occurred in another (awaiting repair). At a median follow-up of 15 months, there was no mortality among the Group 1 patients and all except for 1 patient were symptom free. There were 2 early deaths (3.3%) in the Group 2 patients. Two-ventricular repair, although surgically challenging, should be considered in all patients with two functional ventricles who come for Fontan completion. Comprehensive preoperative imaging and meticulous planning helps in identifying suitable candidates.


Patient health questionnaire for school-based depression screening among Chinese adolescents.
The aim of this study was to determine the reliability and validity of a Chinese version of the Patient Health Questionnaire-9 item (PHQ-9) and its 2 subscales (1 item and 2 items) for the screening of major depressive disorder (MDD) among adolescents in Taiwan. A total of 2257 adolescents were recruited from high schools in Taipei. The participants completed assessments including demographic information, the Chinese version of the PHQ-9, and the Rosenberg Self-Esteem Scale, and data on the number of physical illnesses and mental health service utilizations were recorded. Among them, 430 were retested using the PHQ-9 within 2 weeks. Child psychiatrists interviewed a subsample of the adolescents (n = 165) using the Kiddie-Schedule for Affective Disorder and Schizophrenia Epidemiological Version as the criterion standard. The PHQ-9 had good internal consistency (α = 0.84) and acceptable test-retest reliability (0.80). The participants with higher PHQ-9 scores were more likely to have MDD. Principal component factor analysis of the PHQ-9 yielded a 1-factor structure, which accounted for 45.3% of the variance. A PHQ-9 score ≥15 had a sensitivity of 0.72 and a specificity of 0.95 for recognizing MDD. The area under the receiver operating characteristic curve was 0.90. The screening accuracy of the 2 subscales was also satisfactory, with a Patient Health Questionnaire-2 item cutoff of ≥3 being 94.4% sensitive and 82.5% specific and a Patient Health Questionnaire-1 item cutoff of ≥2 being 61.1% sensitive and 87.7% specific. The PHQ-9 and its 2 subscales appear to be reliable and valid for detecting MDD among ethnic Chinese adolescents in Taiwan.


The emerging face of the HIV epidemic in the Middle East and North Africa.
A volume of quality HIV data has materialized recently in the Middle East and North Africa (MENA). This review provides a thematic narrative of the patterns of HIV infection transmission in this region in light of these data. Tens of integrated bio-behavioral surveillance surveys among hard-to-reach key populations at higher risk have been conducted in MENA in the recent years. Many of the studies reported appreciable and growing HIV prevalence. A few studies found alarming prevalence of as much as 87.2% HIV prevalence among people who inject drugs in Tripoli, Libya. The discovery of these hitherto hidden epidemics was unsettling to some authorities after years in which the importance of a focus on HIV prevention among key populations was not recognized. The new data from MENA indicate growing HIV epidemics among key populations across the region. There is heterogeneity, however, as to which key populations are affected and in what proportions in different countries. In a few countries, HIV appears to affect only one key population and often there is substantial geographical heterogeneity in HIV transmission. Data are indicative of a growing HIV disease burden in this part of the globe, in contrast with the declining epidemics in most other regions.


Diagnosing Disseminated Nocardiosis in a Patient With COVID-19 Pneumonia.
Signs and symptoms of atypical pneumonia include fever, shortness of breath, cough, and chest pain. During the coronavirus disease 2019 (COVID-19) pandemic, identifying other causes of febrile respiratory illness in patients who tested positive for COVID-19 has been very challenging. Concerns over infecting healthcare personnel and other patients can impede further evaluations like bronchial lavage, lung biopsies, and other invasive tests. A very high index of suspicion, perhaps unreasonably so, is required to perform invasive tests to investigate alternative possible causes of the illness. We present the case of a 63-year-old man who presented to the hospital with dyspnea. Chest X-ray demonstrated a consolidation in the left lower lobe lung field with a possible underlying mass, and the patient tested positive for COVID-19. He received the standard treatment for COVID pneumonia at the time in our institution (remdesivir and dexamethasone), empiric antibiotics for community-acquired pneumonia, and was eventually discharged home with supplemental oxygen. Several days later, the patient returned to the hospital again with worsening dyspnea and was readmitted. Persistent illness and worsening imaging prompted bronchoscopy. The bronchoscopy showed narrowing of the airway in the left upper lobe, and Nocardia asteroides was isolated from bronchial aspirate. The isolation of Nocardia prompted an investigation for central nervous system involvement with an magnetic resonance imaging (MRI) of the head. The MRI demonstrated multiple bilateral ring-enhancing lesions in the brain. To our knowledge, this is the first reported case of disseminated nocardiosis superimposed on COVID-19 pneumonia.


DeepH-DTA: Deep Learning for Predicting Drug-Target Interactions: A Case Study of COVID-19 Drug Repurposing.
The rapid spread of novel coronavirus pneumonia (COVID-19) has led to a dramatically increased mortality rate worldwide. Despite many efforts, the rapid development of an effective vaccine for this novel virus will take considerable time and relies on the identification of drug-target (DT) interactions utilizing commercially available medication to identify potential inhibitors. Motivated by this, we propose a new framework, called DeepH-DTA, for predicting DT binding affinities for heterogeneous drugs. We propose a heterogeneous graph attention (HGAT) model to learn topological information of compound molecules and bidirectional ConvLSTM layers for modeling spatio-sequential information in simplified molecular-input line-entry system (SMILES) sequences of drug data. For protein sequences, we propose a squeezed-excited dense convolutional network for learning hidden representations within amino acid sequences; while utilizing advanced embedding techniques for encoding both kinds of input sequences. The performance of DeepH-DTA is evaluated through extensive experiments against cutting-edge approaches utilising two public datasets (Davis, and KIBA) which comprise eclectic samples of the kinase protein family and the pertinent inhibitors. DeepH-DTA attains the highest Concordance Index (CI) of 0.924 and 0.927 and also achieved a mean square error (MSE) of 0.195 and 0.111 on the Davis and KIBA datasets respectively. Moreover, a study using FDA-approved drugs from the Drug Bank database is performed using DeepH-DTA to predict the affinity scores of drugs against SARS-CoV-2 amino acid sequences, and the results show that that the model can predict some of the SARS-Cov-2 inhibitors that have been recently approved in many clinical studies.


Diabetic macular edema: what is focal and what is diffuse?
To review the available information on classification of diabetic macular edema (DME) as focal or diffuse. Interpretive essay. Literature review and interpretation. The terms focal diabetic macular edema and diffuse diabetic macular edema frequently are used without clear definitions. Published definitions often use different examination methods and often are inconsistent. Evaluating published information on the prevalence of focal and diffuse DME, the responses of focal and diffuse DME to treatments, and the importance of focal and diffuse DME in assessing prognosis is hindered because the terms are used inconsistently. A newer vocabulary may be more constructive, one that describes discrete components of the concepts such as extent and location of macular thickening, involvement of the center of the macula, quantity and pattern of lipid exudates, source of fluorescein leakage, and regional variation in macular thickening and that distinguishes these terms from the use of the term focal when describing one type of photocoagulation technique. Developing methods for assessing component variables that can be used in clinical practice and establishing reproducibility of the methods are important tasks. Little evidence exists that characteristics of DME described by the terms focal and diffuse help to explain variation in visual acuity or response to treatment. It is unresolved whether a concept of focal and diffuse DME will prove clinically useful despite frequent use of the terms when describing management of DME. Further studies to address the issues are needed.


Sex frequency and sex planning among men who have sex with men in Bangkok, Thailand: implications for pre- and post-exposure prophylaxis against HIV infection.
Daily HIV antiretroviral pre-exposure prophylaxis (PrEP) is being evaluated in clinical trials among men who have sex with men (MSM). However, daily PrEP may not be congruent with sexual exposure profiles of MSM. Here, we investigate sex frequency and sex planning to identify and inform appropriate PrEP strategies for MSM. We evaluated sex frequency and sex planning in a cohort of HIV-negative MSM in Bangkok, Thailand. Chi-squared test was used to compare reports of sex on different weekdays; logistic regression was used to identify predictors of sex frequency and sex planning. Of 823 MSM (with a mean age of 28.3 years), 86% reported having sex on two days per week or less, and 65% reported their last sex to have been planned. Sex on the weekend (approximately 30%) was more often reported than sex on weekdays (approximately 23%). In multivariate analysis, use of alcohol, erectile dysfunction drugs, group sex, sex with a foreigner, buying and selling sex, and a history of HIV testing were associated with having sex on three days or more per week. Being aged 22 to 29 years, not identifying as homosexual, having receptive anal intercourse, and not engaging in group sex were associated with unplanned sex. Intermittently dosed PrEP (as opposed to daily) may be a feasible HIV prevention strategy and should be considered for evaluation in clinical trials. Risk factors for sex frequency and sex planning may help to identify those in need for daily PrEP and those who may not be able to take a timely pre-exposure dose.


Impact of a financial risk-sharing scheme on budget-impact estimations: a game-theoretic approach.
As part of the process of updating the National List of Health Services in Israel, health plans (the 'payers') and manufacturers each provide estimates on the expected number of patients that will utilize a new drug. Currently, payers face major financial consequences when actual utilization is higher than the allocated budget. We suggest a risk-sharing model between the two stakeholders; if the actual number of patients exceeds the manufacturer's prediction, the manufacturer will reimburse the payers by a rebate rate of α from the deficit. In case of under-utilization, payers will refund the government at a rate of γ from the surplus budget. Our study objective was to identify the optimal early estimations of both 'players' prior to and after implementation of the risk-sharing scheme. Using a game-theoretic approach, in which both players' statements are considered simultaneously, we examined the impact of risk-sharing within a given range of rebate proportions, on players' early budget estimations. When increasing manufacturer's rebate α to be over 50 %, then manufacturers will announce a larger number, and health plans will announce a lower number of patients than they would without risk sharing, thus substantially decreasing the gap between their estimates. Increasing γ changes players' estimates only slightly. In reaction to applying a substantial risk-sharing rebate α on the manufacturer, both players are expected to adjust their budget estimates toward an optimal equilibrium. Increasing α is a better vehicle for reaching the desired equilibrium rather than increasing γ, as the manufacturer's rebate α substantially influences both players, whereas γ has little effect on the players behavior.


Host and Cropping System Shape the Fusarium Population: 3ADON-Producers Are Ubiquitous in Wheat Whereas NIV-Producers Are More Prevalent in Rice.
In recent years, Fusarium head blight (FHB) outbreaks have occurred much more frequently in China. The reduction of burning of the preceding crop residues is suggested to contribute to more severe epidemics as it may increase the initial inoculum. In this study, a large number of Fusarium isolates was collected from blighted wheat spikes as well as from rice stubble with perithecia originating from nine sampling sites in five provinces in Southern China. Fusarium asiaticum dominated both wheat and rice populations, although rice populations showed a higher species diversity. Chemotype analysis showed that rice is the preferred niche for NIV mycotoxin producers that were shown to be less virulent on wheat. In contrast, 3ADON producers are more prevalent on wheat and in wheat producing areas. The 3ADON producers were shown to be more virulent on wheat, revealing the selection pressure of wheat on 3ADON producers. For the first time, members of the Incarnatum-clade of FusariumIncarnatum-Equiseti Species Complex (FIESC) were found to reproduce sexually on rice stubble. The pathogenicity of FIESC isolates on wheat proved very low and this may cause the apparent absence of this species in the main wheat producing provinces. This is the first report of the Fusarium population structure including rice stubble as well as a direct comparison with the population on wheat heads in the same fields. Our results confirm that the perithecia on rice stubble are the primary inoculum of FHB on wheat and that cropping systems affect the local Fusarium population.


Pediatric congenital heart diseases: Patterns of presentation to the emergency department of a tertiary care hospital.
To observe presentation of Pediatric congenital cardiac defects to the Emergency Department (ED) of a tertiary care hospital in Pakistan. This is a retrospective chart review of patients under the age of 16 years with congenital cardiac defects presenting to the Emergency Department of Aga Khan University Hospital over a period of eighteen months, from January 2012 to June 2013. Study population was divided into two groups; first group constituted children with undiagnosed congenital cardiac defects, whereas second group constituted children with diagnosed congenial cardiac defects presented to ED. In previously diagnose cases each visit was counted as a separate encounter. Out of 133 children, 44 (33.5%) were diagnosed congenital cardiac disease for the first time (Group-1) in ED, while 89 (66.5%) children were diagnosed cases of congenital heart disease (Group-2). Among Group-1; main reasons for ED visits were cyanosis, cardiac failure, murmur evaluation and cardiogenic shock where as in Group-2; main presentations were cardiac failure, hyper cyanotic spells, gastroenteritis, lower respiratory tract infection, and post-operative issues. There were total 13 deaths. High index of suspicion is necessary for early diagnosis and management of children with congenital heart disease in the pediatric emergency department.


A comparison of the antigenicity of soya-bean-based infant formulas.
1. The antigenicity of four soya-bean-based infant formulas (Prosobee powder, Prosobee liquid concentrate (Mead Johnson, Uxbridge, Middx), Wysoy (Wyeth, Maidenhead, Berks) and Formula S (Cow and Gate, Trowbridge, Wilts] was measured by enzyme-linked immunosorbent assays (ELISAs) specific for glycinin and beta-conglycinin. Results were compared with in vivo assessments of antigenicity using guinea-pigs, rabbits and calves. 2. The levels of antigenic glycinin and beta-conglycinin in Wysoy and Formula S were below the limits of detection of the ELISA. Both these proteins were detected in Prosobee powder and Prosobee liquid concentrate with the highest levels, especially for glycinin, being present in Prosobee powder. 3. Wysoy was sufficiently antigenic to evoke a soya-bean-specific serum antibody response in rabbits injected with this formula emulsified in complete Freunds adjuvant. A significantly greater response was obtained when rabbits were similarly injected with Prosobee powder. 4. The formulas varied in their ability to sensitize guinea-pigs for both anaphylaxis and antibody production when given orally, although the differences were not statistically significant. Prosobee powder appeared to be the most antigenic and Formula S the least, with Prosobee liquid concentrate and Wysoy being intermediate. 5. Similar variations in antigenicity were observed when Prosobee powder, Wysoy and Formula S were fed to soya-bean-sensitive calves. These formulas were all capable of provoking intestinal disturbances (seen as increased ileal flow-rate, decreased small intestinal transit time and decreased nitrogen absorption) but the most severe reactions were seen when Prosobee powder was fed and the least with Formula S. 6. Thus the four soya-bean-based infant formulas showed considerable differences in antigenicity. In vivo studies using guinea-pigs, rabbits and calves were in good agreement and broadly correlated with the immunochemical assessment of antigenicity. However, the in vitro and in vivo results did not correspond exactly and levels of glycinin and beta-conglycinin below the limit of detection by ELISA could evoke an immune response in the different animal species. We believe that these variations in antigenicity of different commercial products prepared from isolated soya-bean protein may be important when interpreting the results from studies of the development of allergy in infants given soya-bean-based formulas.


Out-of-pocket expenditure on health care by Australian mothers: Lessons for maternal universal health coverage from a long-established system.
Designing effective universal health care systems has challenges, including the use of patient co-payments and the role of the public and private systems. This study sought to quantify the total amount of out-of-pocket fees incurred by women who gave birth in private and public hospitals within Australia-a country with universal health coverage-and assess the impact that variation in birth type has on out-of-pocket fees. Data came from a linked administrative data set of all women who gave birth in the Australian state Queensland between July 1, 2012, and June 30, 2015, plus their resultant children. Propensity score matching was used to create two similar cohorts of women who gave birth in private and public hospitals. The mean total out-of-pocket fees for care from conception to the child's first birthday was $2813 (±2683 standard deviation) and $623 (±1202) for women who gave birth in private and public hospitals, respectively. Total fees were higher in both public and private hospitals for women who had a cesarean birth ($716 [±1419] and $3010 [±2988]) than for women who had a vaginal birth without instruments ($556 [±1044] and $2560 [±2284]). Australia's strong policy incentives for women to take out private health insurance are leaving women with large out-of-pocket costs. This should hold important lessons for other countries implementing a universal health care system, to ensure that using a combination of public and private practitioners does not undermine the intention of universal care.


Electrical stimulation to identify neural elements on the heart: their role in atrial fibrillation.
Anesthetized dogs were subjected to a right then left thoracotomy. Two modes of electrical stimulation were used to activate ganglionated plexi (GP) on the epicardium of the atria: (1) Near the base of each pulmonary vein (PV), trains of high frequency stimuli (HFS) were coupled to each atrial paced beat so as to fall within the refractory period to achieve nerve stimulation without atrial excitation; and (2) Continuous HFS was applied via plaque electrodes sutured to epicardial fat pads (containing a GP) near the right superior (RS) and left superior (LS) PVs. The chest was then closed. An ablation catheter, inserted percutaneously, was positioned fluoroscopically in the right atrium across from the epicardial plaque electrode near the RSPV. Transeptal puncture was used to place an ablation catheter at the LSPV-left atrial junction. HFS applied to each of the epicardial fat pads induced atrial fibrillation (AF) and also caused high grade AV block due to a strong parasympathetic effect on the AV node. Radiofrequency ablation from the right and left atrial endocardium abolished the vagal response to HFS delivered to the plaque electrodes on the fat pads close to the RSPV and LSPV, respectively. Sixty (60) patients with paroxysmal or persistent AF underwent PV antrum isolation (27 patients) or PV antrum isolation plus left atrial GP ablation (33 patients). Endocardial HFS at the border of the PV antra near the 4 GPs produced AF and high grade AV block (vagal response) during AF. RFA at these sites abolished the vagal response. Testing in a small number of patients with very short follow-up suggests that adding GP ablation to PV antrum isolation may increase ablation success (absence of AF recurrence) from 70% to 91%. These basic and clinical studies suggest that localized cardiac autonomic ganglia (GPs) may play a critical role in the initiation and maintenance of AF.


Molecular testing (strand displacement assay) for identification of urethral gonorrhoea in men: can it replace culture as the gold standard?
The objective was to evaluate the performance of Becton Dickinson's BD Probe Tec(TM) (BDPT) strand displacement amplification (SDA) test for the detection of Neisseria gonorrhoeae on urethral specimens from men with urethritis compared with conventional culture and to show that SDA improves the diagnostic yield of gonorrhoea infections (GC). Anonymized retrospective testing of stored urethral swab samples from men attending genitourinary services in East London was performed using SDA. The prevalence of GC culture positive infections in this sample was 20/152 (13%). The sensitivity, specificity, positive predictive value and negative predictive value for the BDPT-SDA system compared with culture were 100%, 95%, 77% and 100%, respectively. In this study population, the BDPT-SDA assay was a highly sensitive and specific test for the diagnosis of N. gonorrhoeae from urethral swabs in men. Therefore, SDA can be used to complement culture in the diagnosis of N. gonorrhoeae infection. No ethics committee approval was obtained as all samples were anonymized.


Older versus younger patients with metastatic adenocarcinoma of the esophagus, gastroesophageal junction, and stomach: a pooled analysis of eight consecutive North Central Cancer Treatment Group (NCCTG) trials.
Whether elderly patients with metastatic esophageal, gastroesophageal, and gastric cancer do as well with chemotherapy as their younger counterparts was investigated in this pooled analysis. In total, 367 patients from 8 consecutive, first-line trials were included: i) etoposide + cisplatin; ii) 5-fluorourucil + leucovorin; iii) 5-fluorouracil + levamisole; iv) irinotecan; v) docetaxel + irinotecan; vi) oxaliplatin + capecitabine; vii) docetaxel + capecitabine; and viii) bortezomib + paclitaxel + carboplatin. One hundred and fifty-four (42%) patients were > or =65 years old (range: 65-86), and 213 younger (range: 20-64). Elderly patients had worse performance scores (2-3): 19 vs. 8% (p<0.0001). Rates of grade 3+ adverse events across all chemotherapy cycles in univariate and multivariate analyses (adjusted for gender, performance score, and stratified by individual study) were higher among elderly patients. Rates of neutropenia, fatigue, infection, and stomatitis in elderly vs. younger patients were 31 vs. 29% (p=0.02 by multivariate analyses); 15 vs. 5% (p=0.01); 9 vs. 4% (p=0.03); 6 vs. 1% (p=0.04). In contrast, duration of chemotherapy, overall survival, and progression-free survival were comparable. Although age should not preclude trial entry, these adverse event rates suggest a need to develop more tolerable regimens for older patients with these malignancies.


[Liquid lipid crystals-induced arthritis].
Liquid lipid crystals-induced arthritis crystals are a frequent cause of arthritis. They are disclosed by the microscopic examination of the synovial fluid. They are usually made of monosodium urate, calcium pyrophosphate or apatite. Liquid lipid crystals have a spherical shape with an aspect of Maltese crosses and a positive birefringence. They are sometimes observed outside the leukocytes in rheumatoid or post-traumatic effusions. In some cases, they constitute the solely cause explaining the arthropathy. Then, they are numerous and located in and outside the cells. This observation reports on the case of a 50 year-old woman having developed this type of arthritis related to a right knee hemarthrosis. Awareness of this pathology may be useful because its evolution is usually good with a non-steroidal anti-inflammatory drug or a local steroid infiltration after exclusion of an infectious origin of the effusion.


MMSET is highly expressed and associated with aggressiveness in neuroblastoma.
MMSET (WHSC1/NSD2) is a SET domain-containing histone lysine methyltransferase the expression of which is deregulated in a subgroup of multiple myelomas with the t(4;14)(p16;q32) translocation associated with poor prognosis. Recent studies have shown that MMSET mRNA levels are increased in other tumor types as well. We have carried out immunohistochemical staining of tissue microarrays and found that MMSET protein is frequently and highly expressed in neuroblastoma (MMSET positive in 75% of neuroblastomas, n = 164). The expression level of MMSET in neuroblastomas was significantly associated with poor survival, negative prognostic factors, and metastatic disease. Moreover, a subset of neuroblastomas for which pre- and postchemotherapy biopsies were available displayed a strong decrease in MMSET protein levels after chemotherapy. In agreement with neuroblastomas becoming more differentiated after treatment, we show that retinoic acid-induced differentiation of human neuroblastoma cells in vitro also leads to a strong decrease in MMSET levels. Furthermore, we show that the high levels of MMSET in normal neural progenitor cells are strongly downregulated during differentiation. Importantly, we show that MMSET is required for proliferation of neuroblastoma cells and brain-derived neural stem cells. Taken together, our results suggest that MMSET is implicated in neuroblastomagenesis possibly by supporting proliferation of progenitor cells and negatively regulating their differentiation. In this respect, MMSET might be a strong candidate therapeutic target in a subset of neuroblastomas with unfavorable prognosis.


Sox2 and Mitf cross-regulatory interactions consolidate progenitor and melanocyte lineages in the cranial neural crest.
The cellular origin and molecular mechanisms regulating pigmentation of head and neck are largely unknown. Melanocyte specification is controlled by the transcriptional activity of Mitf, but no general logic has emerged to explain how Mitf and progenitor transcriptional activities consolidate melanocyte and progenitor cell fates. We show that cranial melanocytes arise from at least two different cellular sources: initially from nerve-associated Schwann cell precursors (SCPs) and later from a cellular source that is independent of nerves. Unlike the midbrain-hindbrain cluster from which melanoblasts arise independently of nerves, a large center of melanocytes in and around cranial nerves IX-X is derived from SCPs, as shown by genetic cell-lineage tracing and analysis of ErbB3-null mutant mice. Conditional gain- and loss-of-function experiments show genetically that cell fates in the neural crest involve both the SRY transcription factor Sox2 and Mitf, which consolidate an SCP progenitor or melanocyte fate by cross-regulatory interactions. A gradual downregulation of Sox2 in progenitors during development permits the differentiation of both neural crest- and SCP-derived progenitors into melanocytes, and an initial small pool of nerve-associated melanoblasts expands in number and disperses under the control of endothelin receptor B (Ednrb) and Wnt5a signaling.


[A new method in the determination of individual delay time in bolus application in spiral CT].
Helical-CT examinations, particularly CT angiography, require precise timing between the examination procedure and the individual dynamics of contrast medium distribution in the arteriovenous system of the patient. The necessary delay between contrast medium injection and onset of has usually been either estimated or determined by means of an additional helical CT examination. The present paper introduces a new technique which allows bolus analysis without an additional scan. Prior to a CT angiography with 10 patients, two techniques for bolus analysis (BA) were compared. Prior to diagnostic contrast medium enhanced examination, a native (BA I) and a dynamic (BA II) examination were performed. Ten seconds prior to the start of each examination, a 10 ml test bolus was applied with an injection flow rate of 3 ml/s. Both examinations lasted for 30 s. During BA I, increase in attenuation in the aorta was compared at different sites, during BA II consistently at the same site. Comparison of the individual peak times yielded a coefficient of correlation of r = 0.926. The median value for BA I was 18.4 +/- 5.4 s and 19.2 +/- 4.5 s for BA II. The difference between measurement of peak time was 1.2 +/- 1.16 s. This modified technique for bolus analysis during the primary native scan of the upper abdominal organs permits calculation of the required individual delay time between contrast medium application and scan start: no additional examination is required and the method can be performed with any helical CT unit.


Self-monitoring of blood glucose in patients with diabetes who do not use insulin--are guidelines evidence-based?
To evaluate if clinical practice guideline recommendations regarding self-monitoring of blood glucose in patients with diabetes not using insulin follow the principles of evidence-based medicine. After a search from 1999 to 2011, 18 clinical practice guidelines were included. Recommendations regarding self-monitoring of blood glucose were graded on a scale from one (strongly against self-monitoring) to four (strongly in favour of self-monitoring) and compared with the similarly graded conclusions of systematic reviews that were cited by the clinical practice guidelines. We also investigated how clinical practice guideline characteristics, for example funding sources, and quality of references cited could be related to the guideline recommendations. The clinical practice guidelines cited in total 15 systematic reviews, 14 randomized controlled trials, 33 non-randomized controlled trials papers and 18 clinical practice guidelines or position statements. The clinical practice guideline recommendations had an average grade of 3.4 (range 2.0-4.0). Higher grades were seen for clinical practice guidelines that acknowledged industry funding (mean value 4.0) or were issued by organizations depending on private funding (mean value 3.6 vs. 3.0 for governmental funding). The conclusions of the 15 systematic reviews had a mean grade of 2.2 (range 1.0-3.8). Systematic reviews with low grades were less cited. In total, 21 randomized controlled trials were included in the systematic reviews. Approximately half of these evaluated an educational intervention where the effect of self-monitoring of blood glucose could not be clearly isolated. Clinical practice guidelines were more in favour of self-monitoring use than the systematic reviews that were cited. The citation practice was non-systematic and industry funding seemingly led to a more positive attitude towards use of self-monitoring of blood glucose.


Ochratoxin A as a potential etiologic factor in endemic nephropathy: lessons from toxicity studies in rats.
Various reports suggest that chronic dietary exposure to ochratoxin A (OTA), a mycotoxin frequently detected in various food items may be linked to the pathogenesis of endemic nephropathy, a chronic tubulointerstitial kidney disease which occurs in geographically limited areas of the Balkan region. OTA is a potent nephrotoxin and renal carcinogen. However, the pathological lesions observed in kidneys of rats treated with OTA appear be rather different from the clinical and pathological characteristics of endemic nephropathy. Moreover, increasing evidence suggests that OTA does not bind to DNA but induces tumors by an epigenetic, thresholded mechanism. This implies that there is a dose below which no adverse health effects are expected to occur. Based on food consumption data and OTA serum concentrations, it appears that human exposure - even in areas with relatively high dietary exposure to OTA such as endemic villages - is several orders of magnitude below doses known to cause nephrotoxicity and tumor formation in laboratory animals. While it is undoubtedly important to encourage prevention of food contamination by OTA and other mycotoxins, these observations suggest that OTA is not likely to be an etiological factor involved in BEN and indicate a need to search for new clues for the etiology of this endemic kidney disease.


Insectivory of savanna chimpanzees (Pan troglodytes verus) at Fongoli, Senegal.
Little is known about the behavior of chimpanzees living in savanna-woodlands, although they are of particular interest to anthropologists for the insight they can provide regarding the ecological pressures affecting early hominins living in similar habitats. Fongoli, Senegal, is the first site where savanna chimpanzees have been habituated for observational data collection and is the hottest and driest site where such observation of chimpanzees occurs today. Previously, indirect evidence suggested these chimpanzees consumed termites throughout the year, an unusual occurrence for western and eastern chimpanzees. Although meat eating by chimpanzees continues to receive much attention, their use of invertebrate prey has received less emphasis in scenarios of hominin evolution. Here, we further examine the invertebrate diet of Fongoli chimpanzees using direct observational methods and accounting for potential environmental influences. Termite feeding positively correlated with high temperatures. Fongoli chimpanzees spend more time obtaining termites than any other chimpanzee population studied, and this extensive insectivory contributes to the list of distinctive behaviors they display relative to chimpanzees living in more forested habitats. We suggest that savanna chimpanzees at Fongoli differ significantly from chimpanzees elsewhere as a result of the selective pressures characterizing their harsh environment, and this contrast provides an example of a viable referential model for better understanding human evolution. Specifically, our results support the hypotheses that invertebrate prey may have figured more prominently into the diet of early hominins in similar habitats, especially given that invertebrates are an important source of protein and other essential nutrients in a highly seasonal environment.


Structural Immunology of Complement Receptors 3 and 4.
Complement receptors (CR) 3 and 4 belong to the family of beta-2 (CD18) integrins. CR3 and CR4 are often co-expressed in the myeloid subsets of leukocytes, but they are also found in NK cells and activated T and B lymphocytes. The heterodimeric ectodomain undergoes considerable conformational change in order to switch the receptor from a structurally bent, ligand-binding in-active state into an extended, ligand-binding active state. CR3 binds the C3d fragment of C3 in a way permitting CR2 also to bind concomitantly. This enables a hand-over of complement-opsonized antigens from the cell surface of CR3-expressing macrophages to the CR2-expressing B lymphocytes, in consequence acting as an antigen presentation mechanism. As a more enigmatic part of their functions, both CR3 and CR4 bind several structurally unrelated proteins, engineered peptides, and glycosaminoglycans. No consensus motif in the proteinaceous ligands has been established. Yet, the experimental evidence clearly suggest that the ligands are primarily, if not entirely, recognized by a single site within the receptors, namely the metal-ion dependent adhesion site (MIDAS). Comparison of some recent identified ligands points to CR3 as inclined to bind positively charged species, while CR4, by contrast, binds strongly negative-charged species, in both cases with the critical involvement of deprotonated, acidic groups as ligands for the Mg2+ ion in the MIDAS. These properties place CR3 and CR4 firmly within the realm of modern molecular medicine in several ways. The expression of CR3 and CR4 in NK cells was recently demonstrated to enable complement-dependent cell cytotoxicity toward antibody-coated cancer cells as part of biological therapy, constituting a significant part of the efficacy of such treatment. With the flexible principles of ligand recognition, it is also possible to propose a response of CR3 and CR4 to existing medicines thereby opening a possibility of drug repurposing to influence the function of these receptors. Here, from advances in the structural and cellular immunology of CR3 and CR4, we review insights on their biochemistry and functions in the immune system.


Links between analysis of surrogate endpoints and endogeneity.
There has been substantive interest in the assessment of surrogate endpoints in medical research. These are measures that could potentially replace 'true' endpoints in clinical trials and lead to studies that require less follow-up. Recent research in the area has focused on assessments using causal inference frameworks. Beginning with a simple model for associating the surrogate and true endpoints in the population, we approach the problem as one of endogenous covariates. An instrumental variables estimator and general two-stage algorithm are proposed. Existing surrogacy frameworks are then evaluated in the context of the model. In addition, we define an extended relative effect estimator as well as a sensitivity analysis for assessing what we term the treatment instrumentality assumption. A numerical example is used to illustrate the methodology.


Dioxygen-derived radicals in biological systems.
Three instances of the involvement of dioxygen-derived radicals in biological systems are considered. The first concerns the formation of radicals in the haemolytic reactions induced by treatment of erythrocytes by phenylhydrazine, as an example of the so-called 'oxidant drugs'. The evidence for the formation of phenyl radicals is considered and their origin in the oxidation of phenylhydrazine by a ferryl derivative of haemoglobin postulated. The relevance to the formation of phenylated iron and porphyrin species is described. It is suspected that many instances of oxidative damage to cellular systems result from the coincidence of unsequestered redox-active metal ions (particularly those of iron and copper), reductants, and dioxygen. As an example, the damage to hepatocytes, grown in a culture medium containing cysteine, is described. The formation of radical species derived from dioxygen during the respiratory burst associated with phagocytosis is discussed. A new electrochemical method of detecting the superoxide ion produced during the respiratory burst is described. Particular emphasis is placed on the relation between the production of radical species such as the hydroxyl radical and the superoxide ion, and the extent of phagocytosis.


A multimodal metabolomics approach using imaging mass spectrometry and liquid chromatography-tandem mass spectrometry for spatially characterizing monoterpene indole alkaloids secreted from roots.
Plants release specialized (secondary) metabolites from their roots to communicate with other organisms, including soil microorganisms. The spatial behavior of such metabolites around these roots can help us understand roles for the communication; however, currently, they are unclear because soil-based studies are complex. Here, we established a multimodal metabolomics approach using imaging mass spectrometry (IMS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) to spatially assign metabolites under laboratory conditions using agar. In a case study using Catharanthus roseus, we showed that 58 nitrogen (N)-containing metabolites are released from the roots into the agar. For the metabolite assignment, we used 15N-labeled and non-labeled LC-MS/MS data, previously reported. Four metabolite ions were identified using authentic standard compounds as derived from monoterpene indole alkaloids (MIAs) such as ajmalicine, catharanthine, serpentine, and yohimbine. An alkaloid network analysis using dot products and spinglass methods characterized five clusters to which the 58 ions belong. The analysis clustered ions from the indolic skeleton-type MIAs to a cluster, suggesting that other communities may represent distinct metabolite groups. For future chemical assignments of the serpentine community, key fragmentation patterns were characterized using the 15N-labeled and non-labeled MS/MS spectra.


Effect of soybean hull supplementation to finishing pigs on the emission of noxious gases from slurry.
Ninety six pigs were assigned on the basis of body weight (BW) to one of four dietary treatments (4 pigs per pen and 6 pens per treatment) and fed for 4 weeks. Four 14.85% CP diets were formulated to contain graded levels of soybean hulls at 0, 5, 10, or 15%, respectively. The results showed that treatments did not affect growth performance. Coefficient of total tract apparent digestibility (CTTAD) for dry matter (DM) and blood urea nitrogen (BUN) concentrations were decreased linearly (P < 0.05) with the addition level of soybean hulls. Slurry ammonia nitrogen (NH(3)-N) was not affected with the increased soybean hulls levels, but volatile fatty acids (VFA) were linearly (P < 0.05) increased. Slurry pH and ammonia (NH(3)) emissions were significantly decreased by the addition of soybean hulls (Linear, P < 0.05). Conversely, slurry hydrogen sulphide (H(2)S) emissions exhibited an increase with the addition of soybean hulls (Linear, P < 0.10). Our data indicate that soybean hulls inclusion can decrease slurry pH value and NH(3) emission without any negative influence on growth performance.


Shorter length of stay in the stroke unit: comparison between the 1990s and 2000s.
The process ruling length of stay (LOS) in hospitals is complex, and changes over time in LOS have not been explored. The purpose of the study was to examine differences in LOS, use of and satisfaction with health-related services, and capacity in activities of daily living (ADLs) during the first year post stroke in 2 groups of patients with mild to moderate stroke who received care in the same stroke unit. The patients (1993/96, n=40; 2006/07, n=43) in this study received care in the stroke unit at Karolinska University Hospital, Huddinge, Sweden. Data on LOS and on the use of health-related services were collected from the Stockholm County Council computerized registers. Satisfaction with health related services was assessed using a questionnaire covering different dimensions of care, while ADLs were assessed using Katz Extended Index of ADL. The LOS in the stroke unit was shorter in the 2006-2007 group (median 8 days) compared to the 1993-1996 group (13 days) (P < .001). Both groups were equally satisfied with health-related services received. A larger proportion of patients were independent in ADLs 3 months post stroke in the 2006-2007 group, but no difference was seen at 6 or 12 months post stroke. It seems possible to reduce the number of days spent in the stroke unit after mild to moderate stroke and instead spend days in a rehabilitation unit, and yet achieve similar patient satisfaction and faster recovery in ADL.


The effects of acute scopolamine in geriatric depression.
In an intensive multidrug, multidose study, nine elderly depressed patients were administered 0.1, 0.25, and 0.5 mg of scopolamine hydrobromide, 1 mg of oral lorazepam, and placebo in a double-blind investigation aimed at assessing the status of the central cholinergic nervous system in geriatric depression. Significant cognitive and behavioral effects of scopolamine were observed only at the high dose (0.5 mg), while lower doses and lorazepam showed no significant differences from placebo. Cognitive deficits caused by scopolamine were in the areas of new learning, access to semantic memory, vigilance, and continuous performance. Behavioral effects consisted of activation, restlessness, and anxiety, but there was no significant effect on depressed mood. These results suggest that elderly depressed patients with mild to moderate cognitive impairment seem to be more similar to previously studied elderly controls rather than to patients with Alzheimer's disease in their reaction to short-term cholinergic blockade, and suggest that the cognitive and mood changes often seen in geriatric depression may involve factors other than disturbed muscarinic cholinergic mechanisms.


Complications following thiacetarsamide sodium therapy in Louisiana dogs with naturally-occurring heartworm disease.
Four hundred and sixteen dogs with naturally-occurring heartworm disease were evaluated for complications following thiacetarsamide sodium therapy. Of these, 109 dogs (26.2%) experienced complications. Increased lung sounds was the most commonly seen complication, followed by fever and coughing. In dogs with complications, 83.5% of them presented without clinical evidence of pulmonary thromboembolism or heart failure. There were no statistically significant differences between the age, sex, breed and body size of dogs that experienced complications following thiacetarsamide therapy and dogs that did not. Complications were most frequently seen 5 to 9 days following thiacetarsamide therapy although some dogs experienced initial complications as late as 28 days. Thirty-three of 109 dogs (33.0%) with complications responded to exercise restriction. The remaining 76 dogs with complications prior to or following thiacetarsamide required adjunct drug therapy. Of these, 35 dogs responded favorably to anti-inflammatory doses of prednisolone or prednisone. Five dogs died or were euthanatized because of the complications experienced. Eighteen of 416 dogs (4.3%) presented with clinical evidence of pulmonary thromboembolism or heart failure prior to the thiacetarsamide therapy. All 18 dogs experienced complications in spite of adjunct drug therapy and exercise restriction prior to, during, and following thiacetarsamide therapy. Survival rate following resolution of the thiacetarsamide-induced complications was greater than 98%.


Developmental heptachlor exposure increases susceptibility of dopamine neurons to N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)in a gender-specific manner.
Parkinson's disease (PD) is primarily thought of as a disease of aging. However, recent evidence points to the potential for exposure to xenobiotics during development to increase risk of PD. Here, we report that developmental exposure to the organochlorine pesticide heptachlor alters the dopamine system and increases neurotoxicity in an animal model of PD. Exposure of pregnant mice to heptachlor led to increased levels of the dopamine transporter (DAT) and vesicular monoamine transporter 2 (VMAT2) levels at both the protein and mRNA level in their offspring. Increased DAT and VMAT2 levels were accompanied by alterations of mRNA levels of nuclear transcription factors that control dopamine neuron development and regulate DAT and VMAT2 levels in adulthood. At 12 weeks of age, control and heptachlor-exposed offspring were administered a moderate dose (2 x 10mg/kg) of the parkinsonism-inducing agent MPTP. Greater neurotoxicity as evidenced by a greater loss of striatal dopamine and potentiation of increased levels of glial fibrillary acidic protein and alpha-synuclein was observed in heptachlor-exposed offspring. The neurotoxicity observed was greater in the male offspring than the female offspring, suggesting that males are more susceptible to the long-term effects of developmental heptachlor exposure. These data suggest that developmental heptachlor exposure causes long-term alterations of the dopamine system thereby rendering it more susceptible to dopaminergic damage in adulthood.


Imaging human cerebral pain modulation by dose-dependent opioid analgesia: a positron emission tomography activation study using remifentanil.
Previous imaging studies have demonstrated a number of cortical and subcortical brain structures to be activated during noxious stimulation and infusion of narcotic analgesics. This study used O-water and positron emission tomography to investigate dose-dependent effects of the short-acting mu-selective opioid agonist remifentanil on regional cerebral blood flow during experimentally induced painful heat stimulation in healthy male volunteers. Positron emission tomography measurements were performed with injection of 7 mCi O-water during nonpainful heat and painful heat stimulation of the volar forearm. Three experimental conditions were used during both sensory stimuli: saline, 0.05 microg x kg x min remifentanil, and 0.15 microg x kg x min remifentanil. Cardiovascular and respiratory parameters were monitored noninvasively. Across the three conditions, dose-dependent effects of remifentanil on regional cerebral blood flow were analyzed on a pixel-wise basis using a statistical parametric mapping approach. During saline infusion, regional cerebral blood flow increased in response to noxious thermal stimulation in a number of brain regions as previously reported. There was a reduction in pain-related activations with increasing doses of remifentanil in the thalamus, insula, and anterior and posterior cingulate cortex. Increasing activation occurred in the cingulofrontal cortex (including the perigenual anterior cingulate cortex) and the periaqueductal gray. Remifentanil induced regional cerebral blood flow increases in the cingulofrontal cortex and periaqueductal gray during pain stimulation, indicating that mu-opioidergic activation modulates activity in pain inhibitory circuitries. This provides direct evidence that opioidergic analgesia is mediated by activation of established descending antinociceptive pathways.


Di-μ 2-acetato-1:2κ (2) O:O';2:3κ (2) O:O'-bis-{μ 2-4,4'-di-chloro-2,2'-[2,2-di-methyl-propane-1,3-diylbis(nitrilo-methanylyl-idene)]diphenolato}-1:2κ (6) O,N,N',O':O,O';2:3κ (6) O,O':O,N,N',O'-tri-cadmium.
In the title linear homo-trinuclear complex, [Cd3(C19H18Cl2N2O2)2(C2H3O2)2], the central Cd(II) atom is located on a centre of inversion and has a distorted octa-hedral coordination geometry formed by four O atoms from two bidentate/tetra-dentate Schiff base ligands and two O atoms from two bridging acetate ligands. The coordination geometry of the terminal Cd(II) atom is square-pyramidal with the tetra-dentate part of the ligand in the basal plane and one O atom from an acetate ligand occupying the apical site. The six-membered CdN2C3 ring adopts a chair conformation. The acetate-bridged Cd⋯Cd distance is 3.3071 (2) Å. The crystal structure is stabilized by C-H⋯O hydrogen bonds, which form C(7) chain motifs and give rise to a two-dimensional supra-molecular network structure lying parallel to the ab plane.


Multi-segment spine kinematics: Relationship with dance training and low back pain.
Spine posture, range of motion (ROM) and movement asymmetry can contribute to low back pain (LBP). These variables may have greater impact in populations required to perform repetitive spine movements, such as dancers; however, there is limited evidence to support this. What is the influence of dance and LBP on spinal kinematics? In this cross-sectional study, multi-segment spinal kinematics were examined in 60 female participants, including dancers (n = 21) and non-dancers (n = 39) with LBP (n = 33) and without LBP (n = 27). A nine-camera motion analysis system sampling at 100 Hz was used to assess standing posture, as well as ROM and movement asymmetry for side bend and trunk rotation tasks. A two-way ANOVA was performed for each of the outcome variables to detect any differences between dancers and non-dancers, or individuals with and without LBP. Compared to non-dancers, dancers displayed a flatter upper lumbar angle when standing (p < 0.01, ηp2 = 0.15), and achieved greater frontal plane ROM for the upper lumbar (p = 0.04, ηp2 = 0.08) and lower thoracic (p = 0.02, ηp2 = 0.09) segments. There were no differences between dancers and non-dancers for transverse plane ROM (p > 0.05) or movement asymmetry (p > 0.05). There was no main effect for LBP symptoms on any kinematic measures, and no interaction effect for dance group and LBP on spinal kinematics (p > 0.05). Female dancers displayed a flatter spine posture and increased spine ROM compared to non-dancers for a select number of spine segments and movement tasks. However, the overall number of differences was small, and no relationship was observed between LBP and spinal kinematics. This suggests that these simple, static posture, ROM, and asymmetry measures often used in clinical practice can provide only limited generalisable information about the impact of dance or LBP on spinal kinematics.


Improved Elastic Recovery from ABC Triblock Terpolymers.
The promise of ABC triblock terpolymers for improving the mechanical properties of thermoplastic elastomers is demonstrated by comparison with symmetric ABA/CBC analogs having similar molecular weights and volume fraction of B and A/C domains. The ABC architecture enhances elasticity (up to 98% recovery over 10 cycles) in part through essentially full chain bridging between discrete hard domains leading to the minimization of mechanically unproductive loops. In addition, the unique phase space of ABC triblocks also enables the fraction of hard-block domains to be higher (fhard ≈ 0.4) while maintaining elasticity, which is traditionally only possible with non-linear architectures or highly asymmetric ABA triblock copolymers. These advantages of ABC triblock terpolymers provide a tunable platform to create materials with practical applications while improving our fundamental understanding of chain conformation and structure-property relationships in block copolymers.


A comparison of the osmotic activation of basophils and human lung mast cells.
Basophils and mast cells release histamine in hyperosmolar environments. Osmotic release differs significantly from IgE-dependent activation and may be important in the pathophysiology of anaphylactoid reactions during intravenous infusions of hyperosmolar radiocontrast dyes and of obstructive attacks suffered by asthmatics after exercise. To confirm reported differences in osmotic activation of basophils and lung mast cells, the process was compared systematically in the 2 cell types. Both cells were activated by hyperosmolar mannitol, glucose, sucrose, and NaCl, but histamine release from basophils rose sharply to a maximum at 1,050 mOsm/kg, whereas mast cells released maximally at 700 to 750 mOsm/kg. Release was partially Ca2+-dependent in basophils but was highly Ca2+-dependent in mast cells. Release in mast cells was rapid and essentially complete by 5 min, whereas 45 to 60 min were required in basophils. The temperature optimum in both cell types was 32 degrees C, and release in both was enhanced by drugs that increase intracellular cAMP (PGE2, IBMX, and db cAMP). We conclude that osmotic activation of basophils and mast cells is a nontoxic process that differs significantly from IgE-dependent histamine release. If it does indeed participate in the pathophysiology of human disease, different treatment strategies will be required to modulate this contribution.


Complex sexual courtship displays by luminescent male marine ostracods.
In the western Caribbean Sea, about an hour after the sun sets, a complex and ritualized light show of precise, vertically placed luminescent pulses erupts over shallow grassbeds. These are among the most complex displays known in marine systems. Displays consist of repeated trains of secreted bioluminescent pulses in a specific pattern ejected into the water column as courtship signals by male Vargula annecohenae, which are small (<2 mm) myodocopid ostracod crustaceans. Although these animals display in near darkness, we have used image intensification and infrared videography and three-dimensional analysis in the lab to demonstrate that each luminescent display train, which can be up to 60 cm long, consists of two distinct luminescent and swimming phases. The first, or 'stationary,' phase consists of three (usually) bright, longer pulses placed close together, with the male swimming in a looping pattern. We hypothesize that this pattern acts as an attention-grabbing signal for receptive females. The stationary phase is followed by the 'helical phase,' which consists of about a dozen evenly placed dimmer, shorter pulses secreted by an individual male rapidly spiraling upward in a helical pattern. We hypothesize that this phase, which has very uniform interpulse intervals and distances, helps an approaching female target and intercept the rapidly moving male. Here we provide details of these two phases, and produce a three-dimensional model of a multiply-displaying male.


Analysis of the Legionella longbeachae genome and transcriptome uncovers unique strategies to cause Legionnaires' disease.
Legionella pneumophila and L. longbeachae are two species of a large genus of bacteria that are ubiquitous in nature. L. pneumophila is mainly found in natural and artificial water circuits while L. longbeachae is mainly present in soil. Under the appropriate conditions both species are human pathogens, capable of causing a severe form of pneumonia termed Legionnaires' disease. Here we report the sequencing and analysis of four L. longbeachae genomes, one complete genome sequence of L. longbeachae strain NSW150 serogroup (Sg) 1, and three draft genome sequences another belonging to Sg1 and two to Sg2. The genome organization and gene content of the four L. longbeachae genomes are highly conserved, indicating strong pressure for niche adaptation. Analysis and comparison of L. longbeachae strain NSW150 with L. pneumophila revealed common but also unexpected features specific to this pathogen. The interaction with host cells shows distinct features from L. pneumophila, as L. longbeachae possesses a unique repertoire of putative Dot/Icm type IV secretion system substrates, eukaryotic-like and eukaryotic domain proteins, and encodes additional secretion systems. However, analysis of the ability of a dotA mutant of L. longbeachae NSW150 to replicate in the Acanthamoeba castellanii and in a mouse lung infection model showed that the Dot/Icm type IV secretion system is also essential for the virulence of L. longbeachae. In contrast to L. pneumophila, L. longbeachae does not encode flagella, thereby providing a possible explanation for differences in mouse susceptibility to infection between the two pathogens. Furthermore, transcriptome analysis revealed that L. longbeachae has a less pronounced biphasic life cycle as compared to L. pneumophila, and genome analysis and electron microscopy suggested that L. longbeachae is encapsulated. These species-specific differences may account for the different environmental niches and disease epidemiology of these two Legionella species.


Transbuccal tablets of carbamazepine: formulation, release and absorption pattern.
Tranbsuccal drug administration is an attractive method, as it has several advantages especially with respect to peroral delivery. Here we report: i) the aptitude of carbamazepine (CBZ) to penetrate porcine buccal mucosa and reconstituted human oral (RHO) epithelium; ii) three different tablet formulations for transbuccal administration; iii) the drug release rate from tablets. CBZ permeation through the buccal mucosa was investigated by using two different bi-compartmental open models: Franz cells for porcine buccal mucosa and Transwell diffusion cells system for RHO epithelium. Results, expressed as drug flux (Js) and permeability coefficients (Kp), indicated that CBZ well penetrates the membrane and arrives in the acceptor phase. Js and Kp resulted 7x10(-2) mg/cm2h and 0.23 cm/h for in vitro experiments and 1.81 x 10(-2) mg/cm2h and 4.57 x 10(-2) cm/h for ex vivo experiments. The flux is extensively affected by the membrane thickness. The CBZ release from three different formulations of tablets, prepared with loaded microspheres, loaded matrices, and conventional compressed physical mixture of components was studied. Using the new formulated "non-conventional" tablets prolonged drug release was obtained. Loaded matrix tablets discharged CBZ faster than microsphere tablets (17% and 12% in about 2.5 h respectively). Results indicate the possibility of administering CBZ on buccal mucosa.


Complex human adenoid tissue-based ex vivo culture systems reveal anti-inflammatory drug effects on germinal center T and B cells.
Human immunology research is often limited to peripheral blood. However, there are important differences between blood immune cells and their counterparts residing in secondary lymphoid organs, such as in the case of germinal center (GC) T follicular helper (Tfh) cells and GC B cells. We developed a versatile ex vivo lymphoid organ culture platform that is based on human pharyngeal tonsils (adenoids) and allows for drug testing. We systematically phenotyped Tfh and GC B cell subsets in explant- and suspension cultures using multicolor flow cytometry and cytokine multiplex analysis. Phenotypic changes of certain ex vivo cultured immune cell subsets could be modulated by cytokine addition. Furthermore, we optimized an activation-induced marker assay to evaluate the response to T cell stimulation. We provide proof-of-concept that Tfh and GC B cells could be modulated in these cultures by different anti-inflammatory drugs in unstimulated states and upon activation with vaccine-derived antigens. For example, GC B cells were lost upon CD40L blockade, and clinically approved JAK inhibitors impacted Tfh and GC B cells, including down-regulation of their key transcription factor BCL6. BCL6 regulation was affected by IL-6 signaling in T cells and IL-4 in B cells, respectively. Furthermore, we demonstrated that JAK signaling and TNF signaling contributed to the stimulation-induced activation of tonsil-derived T cells. Our optimized methods, assays, and mechanistic findings can contribute to a better understanding of human GC responses. These insights may be relevant for improving autoimmune disease therapy and vaccination efficacy. This work was supported by a project grant under the joint research cooperation agreement of LMU Munich, LMU University Hospital, and Sanofi-Aventis Deutschland GmbH, as well as by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - Emmy Noether Programme BA 5132/1-1 and BA 5132/1-2 (252623821), SFB 1054 Project B12 (210592381), and SFB 914 Project B03 (165054336).