Meat allergy associated with galactosyl-α-(1,3)-galactose (α-Gal)-Closing diagnostic gaps by anti-α-Gal IgE immune profiling.

BACKGROUND: Glycoproteins and glycolipids of some mammalian species contain the disaccharide galactosyl-α-(1,3)-galactose (α-Gal). It is known that α-Gal is immunogenic in humans and causes glycan-specific IgG and also IgE responses with clinical relevance. α-Gal is part of the IgE-reactive monoclonal therapeutic antibody cetuximab (CTX) and is associated with delayed anaphylaxis to red meat. In this study, different α-Gal-containing analytes are examined in singleplex and multiplex assays to resolve individual sensitization patterns with IgE against α-Gal. METHODS: Three serum groups, α-Gal-associated meat allergy (MA) patients, idiopathic anaphylaxis (IA) patients with suspected MA, and non-meat-allergic healthy control individuals (HC), were analyzed via singleplex allergy diagnostics and a newly established immunoblot diagnostic system. The new dot blot detection system resolved individual IgE sensitization profiles for α-Gal-containing analytes CTX, bovine thyroglobulin (Bos d TG), and human serum albumin (HSA)-conjugated α-Gal. RESULTS: Singleplex allergy diagnostics using the α-Gal analytes CTX and Bos d TG confirms the history of MA patients in 91% and 88% of the cases, respectively. A novel dot blot-based assay system for the detection of IgE against α-Gal reveals individual IgE sensitization profiles for α-Gal-containing analytes. An α-Gal-associated IgE cross-reactivity profile (IgE against CTX, Bos d TG, and HSA-α-Gal) was identified, which is associated with MA. CONCLUSIONS: Detection of individual sensitization patterns with different α-Gal-containing analytes provides the basis for an individual allergy diagnosis for α-Gal-sensitized patients. Higher amounts of α-Gal in pork and beef innards compared to muscle meat as indicated by a higher staining intensity are a plausible explanation for the difference in allergic symptom severity.

[What does pain intensity mean from the patient perspective? : A qualitative study on the patient perspective of pain intensity as an outcome parameter in treatment evaluation and on the interpretability of pain intensity measurements]. / Was bedeutet Schmerzintensität aus Sicht der Patienten? : Eine qualitative Studie zur Patientenperspektive auf Schmerzstärke als Konstrukt in der Therapieevaluation und die Interpretierbarkeit der Schmerzstärkemessung.

BACKGROUND: Pain intensity (PI) is a common outcome parameter in effectiveness studies on interdisciplinary multimodal pain therapy (IMPT), despite the fact that IMPT highlights dealing with rather than reducing chronic pain. Moreover, the measurement of pain intensity as a highly subjective experience is problematic. Patient participation is absolutely essential to examine the relevance of PI as a criterion of treatment success as well as to select/develop suitable measurement methods. METHOD: A qualitative multicenter study was conducted using focus groups with 69 patients (18-77 years; 80% female) at four different IMPT centers in Germany to discuss pain intensity as a therapy outcome parameter in IMPT, as well as the interpretability and feasibility of common measurement methods. RESULTS: The discussions emphasized that PI is a relevant, but not the primary, outcome in IMPT for patients. Patients' statements also demonstrate that there are some problems in measuring PI, for instance with regard to pain attacks. CONCLUSIONS: The focus group discussions suggested that, due to the highly subjective nature of PI, as well as (verbal) inaccuracies and a lack of standardization in common instruments, the measurement of pain intensity is a challenging task. These limitations should be taken into account in future studies.

IκBα glutathionylation and reduced histone H3 phosphorylation inhibit eotaxin and RANTES.

Airway smooth muscle cells (ASMCs) secrete eotaxin and RANTES (regulated on activation, normal T-cell expressed and secreted) in response to tumour necrosis factor (TNF)-α, which is inhibited by the nuclear factor (NF)-κB inhibitor dimethylfumarate (DMF). NF-κB/IκB (inhibitor of NF-κB) glutathionylation and changes in chromatin remodelling can inhibit NF-κB activity. In this study, we determined whether NF-κB/IκB glutathionylation and reduced histone H3 phosphorylation might underlie the inhibitory effect of DMF on NF-κB activity, and eotaxin and RANTES secretion. Primary human ASMCs were treated with DMF, diamide and/or glutathione (GSH) ethylester (OEt) prior to TNF-α stimulation and were subsequently analysed by ELISA, electrophoretic mobility shift assay, immunofluorescence, co-immunoprecipitation or immunoblotting. DMF reduced intracellular GSH and induced IκBα glutathionylation (IκBα-SSG), which inhibited IκBα degradation, NF-κB p65 nuclear entry and NF-κB/DNA binding. In addition, DMF inhibited the phosphorylation of histone H3, which was possibly mediated by the inhibitory effect of DMF on mitogen- and stress-activated protein kinase (MSK)-1. However, p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase MAPK and MAPK phosphatase-1, upstream of MSK-1, were not inhibited by DMF. Importantly, DMF-mediated effects on NF-κB, histone H3, eotaxin and RANTES were reversed by addition of GSH-OEt. Our data suggest that DMF inhibits NF-κB-dependent eotaxin and RANTES secretion by reduction of GSH with subsequent induction of IκBα-SSG and inhibition of histone H3 phosphorylation. Our findings offer new potential drug targets to reduce airway inflammation in asthma.

Dimethylfumarate inhibits NF-{kappa}B function at multiple levels to limit airway smooth muscle cell cytokine secretion.

The antipsoriatic dimethylfumarate (DMF) has been anecdotically reported to reduce asthma symptoms and to improve quality of life of asthma patients. DMF decreases the expression of proinflammatory mediators by inhibiting the transcription factor NF-kappaB and might therefore be of interest for the therapy of inflammatory lung diseases. In this study, we determined the effect of DMF on platelet-derived growth factor (PDGF)-BB- and TNFalpha-induced asthma-relevant cytokines and NF-kappaB activation by primary human asthmatic and nonasthmatic airway smooth muscle cells (ASMC). Confluent nonasthmatic and asthmatic ASMC were incubated with DMF (0.1-100 microM) and/or dexamethasone (0.0001-0.1 microM), NF-kappaB p65 siRNA (100 nM), the NF-kappaB inhibitor helenalin (1 microM) before stimulation with PDGF-BB or TNFalpha (10 ng/ml). Cytokine release was measured by ELISA. NF-kappaB, mitogen and stress-activated kinase (MSK-1), and CREB activation was determined by immunoblotting and EMSA. TNFalpha-induced eotaxin, RANTES, and IL-6 as well as PDGF-BB-induced IL-6 expression was inhibited by DMF and by dexamethasone from asthmatic and nonasthmatic ASMC, but the combination of both drugs showed no glucocorticoid sparing effect in either of the two groups. NF-kappaB p65 siRNA and/or the NF-kappaB inhibitor helenalin reduced PDGF-BB- and TNFalpha-induced cytokine expression, suggesting the involvement of NF-kappaB signaling. DMF inhibited TNFalpha-induced NF-kappaB p65 phosphorylation, NF-kappaB nuclear entry, and NF-kappaB-DNA complex formation, whereas PDGF-BB appeared not to activate NF-kappaB within 60 min. Both stimuli induced the phosphorylation of MSK-1, NF-kappaB p65 at Ser276, and CREB, and all were inhibited by DMF. These data suggest that DMF downregulates cytokine secretion not only by inhibiting NF-kappaB but a wider range of NF-kappaB-linked signaling proteins, which may explain its potential beneficial effect in asthma.

Violent Behavior During Psychiatric Inpatient Treatment in a German Prison Hospital.

Violent behavior in correctional facilities is common and differs substantially in type, target, implication, and trigger. Research on frequency and characteristics of violent behavior in correctional facilities and psychiatric hospitals is limited. Results from recent research suggest that comorbidity of severe mental disorder, personality disorder, and diagnosis of substance abuse is related to a higher risk of violent behavior. In the Berlin prison hospital, a database was created to collect data from all violent incidences (n=210) between 1997 and 2006 and between 2010 and 2016. In a retrospective, case-control study, we analyzed specific socioeconomic data and psychiatric diagnosis and compared the group of prisoners with violent behavior with randomly selected prisoners of the same department without violent behavior (n = 210). Diagnosis of schizophrenia, non-German nationality, no use of an interpreter, no children, and no previous sentence remained significantly associated with the dependent variable violent behavior. There were no significant differences regarding age and legal statuses. Practical implications for clinical work are discussed.

Corrigendum: Violent Behavior During Psychiatric Inpatient Treatment in a German Prison Hospital.

[This corrects the article DOI: 10.3389/fpsyt.2019.00762.].

Multilayer perceptron tumour diagnosis based on chromatography analysis of urinary nucleosides.

Nucleosides in human urine are of interest as a biochemical marker for cancer, acquired immune deficiency syndrome (AIDS) and the whole-body turnover of RNAs. A reversed-phase high-performance liquid chromatography (RP-HPLC) method with photodiode-array detection was used to quantitatively analyze urinary normal and modified nucleosides. 55 persons with malignant tumors of various types, 13 persons with benign tumors and 41 healthy controls were investigated within a clinical intervention study. Artificial neural networks (ANN) have been used as a practical pattern recognition tool to distinguish cancer patients from healthy persons. Using a multilayer perceptron (MPL), a specificity of 85%, and a sensitivity of 97% in differentiation between tumor patients and healthy persons was achieved. The differentiation between benign and malignant tumors had a sensitivity of 60% and a specificity of 84%. These results verify the usefulness of ANN and the RP-HPLC method for tumor recognition in agreement with existing studies.

Anomalous Inner-Gap Structure in Transport Characteristics of Superconducting Junctions with Degraded Interfaces.

Quantitative description of charge transport across tunneling and break-junction devices with novel superconductors encounters some problems not present or not as severe for traditional superconducting materials. In this work, we explain unexpected features in related transport characteristics as an effect of a degraded nanoscaled sheath at the superconductor surface. A model capturing the main aspects of the ballistic charge transport across hybrid superconducting structures with normally conducting nanometer-thick interlayers is proposed. The calculations are based on a scattering formalism taking into account Andreev electron-into-hole (and inverse) reflections at normal metal-superconductor interfaces as well as transmission and backscattering events in insulating barriers between the electrodes. Current-voltage characteristics of such devices exhibit a rich diversity of anomalous (from the viewpoint of the standard theory) features, in particular shift of differential-conductance maxima at gap voltages to lower positions and appearance of well-defined dips instead expected coherence peaks. We compare our results with related experimental data.

Intrinsic and extrinsic pinning in NdFeAs(O,F): vortex trapping and lock-in by the layered structure.

Fe-based superconductors (FBS) present a large variety of compounds whose properties are affected to different extents by their crystal structures. Amongst them, the REFeAs(O,F) (RE1111, RE being a rare-earth element) is the family with the highest critical temperature Tc but also with a large anisotropy and Josephson vortices as demonstrated in the flux-flow regime in Sm1111 (Tc ∼ 55 K). Here we focus on the pinning properties of the lower-Tc Nd1111 in the flux-creep regime. We demonstrate that for H//c critical current density Jc at high temperatures is dominated by point-defect pinning centres, whereas at low temperatures surface pinning by planar defects parallel to the c-axis and vortex shearing prevail. When the field approaches the ab-planes, two different regimes are observed at low temperatures as a consequence of the transition between 3D Abrikosov and 2D Josephson vortices: one is determined by the formation of a vortex-staircase structure and one by lock-in of vortices parallel to the layers. This is the first study on FBS showing this behaviour in the full temperature, field, and angular range and demonstrating that, despite the lower Tc and anisotropy of Nd1111 with respect to Sm1111, this compound is substantially affected by intrinsic pinning generating a strong ab-peak in Jc.

Mammalian expression of the human sex steroid-binding protein of plasma (SBP or SHBG) and testis (ABP). Characterization of the recombinant protein.

A full-length 1,209 bp cDNA encoding the human sex steroid-binding protein of plasma (SBP or SHBG) and testis (ABP) was constructed and expressed in BHK-21 cells. The sequence agrees with the published gene and protein sequences. The cells were found to secrete SBP following transfection and G418r selection. The recombinant protein binds 5 alpha-dihydrotestosterone with a Kd of 0.28 nM. It also binds testosterone and 17 beta-estradiol but not progesterone, estrone or cortisol revealing a steroid-binding specificity identical to that of human SBP. SDS-PAGE patterns are less complex than human SBP and show a monomeric molecular weight of about 43 kDa.

Evidence for a specific recognition site for tiflucarbine on calmodulin.

The putative antidepressant drug tiflucarbine (BAY P 4495) has previously been shown to inhibit calmodulin-dependent cyclic nucleotide phosphodiesterase competitively with respect to calmodulin. In order to determine whether this effect is mediated by a direct interaction with calmodulin, we measured the effects of radiolabelled triflucarbine in a direct ligand binding assay, using agarose-immobilized calmodulin. [3H]Tiflucarbine associated with low micromolar affinity with an apparently homogeneous class of recognition sites on calmodulin-agarose. No binding could be observed on calmodulin-deficient agarose. The effect was specific, saturable and reversible. Triflucarbine was the most potent calmodulin antagonist from a variety of structural analogues examined. The potencies of these derivatives to inhibit calmodulin-stimulated phosphodiesterase significantly correlated with their affinities towards the tiflucarbine binding site on calmodulin. No such correlation was evident when structurally unrelated reference compounds were tested. The association of tiflucarbine with calmodulin thus appears pharmacologically specific and selective and possibly contributes to the potent antidepressant activity of the drug.

Cyclosporins from Tolypocladium terricola.

New natural cyclosporins were isolated from the mycelium of surface cultivated fungus Tolypocladium terricola. The chemical structures of [Leu4] CS and [MeLeu1] CS = cyclosporin-J, were deduced from the NMR and mass spectral data. Biological activity of new cyclosporins is reported based on the proliferative mitogen stimulation test.

Brain serotonin receptors as a target for the putative anxiolytic TVX Q 7821.

In animal behavioral tests of anxiolytic efficacy, TVX Q 7821 was active and equipotent with diazepam, but did not produce muscle relaxation or anti-convulsant effects. The high affinity, specific binding of 3H-TVX Q 7821 to calf hippocampal membranes was displaced by serotonin (5-HT) but not by diazepam. Similarly, unlabeled TVX Q 7821 displaced 3H-5-HT but not 3H-flunitrazepam binding. Since ketanserin (a putative 5-HT2 ligand) was equally weak in displacing labeled 5-HT or TVX Q 7821, TVX Q 7821 may preferentially bind to 5-HT1, receptors.

Reproducibility of HTS-SQUID magnetocardiography in an unshielded clinical environment.

A new technology has been developed which measures the magnetic field of the human heart (magnetocardiogram, MCG) by using high temperature superconducting (HTS) sensors. These sensors can be operated at the temperature of liquid nitrogen without electromagnetic shielding. We tested the reproducibility of HTS-MCG measurements in healthy volunteers. Unshielded HTS-MCG measurements were performed in 18 healthy volunteers in left precordial position in two separate sessions in a clinical environment. The heart cycles of 10 min were averaged, smoothed, the baselines were adjusted, and the data were standardized to the respective areas under the curves (AUC) of the absolute values of the QRST amplitudes. The QRS complexes and the ST-T intervals were used to assess the reproducibility of the two measurements. Ratios (R(QRS), R(STT)) were calculated by dividing the AUC of the first measurement by the ones of the second measurement. The linear correlation coefficients (CORR(QRS), CORR(STT)) of the time intervals of the two measurements were calculated, too. The HTS-MCG signal was completely concealed by the high noise level in the raw data. The averaging and smoothing algorithms unmasked the QRS complex and the ST segment. A high reproducibility was found for the QRS complex (R(QRS)=1.2+/-0.3, CORR(QRS)=0.96+/-0.06). Similarly to the shape of the ECG it was characterized by three bends, the Q, R, and S waves. In the ST-T interval, the reproducibility was considerably lower (R(STT)=0.9+/-0.2, CORR(STT)=0.66+/-0.28). In contrast to the shape of the ECG, a baseline deflection after the T wave which may belong to U wave activity was found in a number of volunteers. HTS-MCG devices can be operated in a clinical environment without shielding. Whereas the reproducibility was found to be high for the depolarization interval, it was considerably lower for the ST segment and for the T wave. Therefore, before clinically applying HTS-MCG systems to the detection of repolarization abnormalities in acute coronary syndromes, further technical development of the systems is necessary to improve the signal-to-noise ratio.

Non-invasive assessment of right and left ventricular volumes 11 to 24 years after corrective surgery on patients with tetralogy of Fallot.

Patients who had corrective surgery for tetralogy of Fallot using a transannular right ventricular outflow tract (TRVOT) patch showed a higher risk of post-operative mortality and reoperation in the long-term follow-up. A total of 642 patients were operated upon for tetralogy of Fallot between 1952 and 1982. Twenty-six patients who survived for more than 10 years were selected for this study. Fifteen randomly selected patients (group 1) without outflow tract patch were compared with 11 patients (group 2) where a transannular right ventricular outflow patch had been used. Right and left ventricular volumes were assessed using combined first-pass and equilibrium radionuclide ventriculography. After administration of 25 mCi of 99mTc-pertechnetate, data were acquired with a gamma camera with a large viewing field and the patient at rest and during exercise. The patients in group 1 were able to manage a workload of 65 +/- 24 W on the bicycle ergometer while the patients in group 2 could only reach a mean maximum of 34 +/- 12 W. Right ventricular end-diastolic volume (RV-EDV) at rest was 198 +/- 67 ml in group 1 and 224 +/- 69 ml in group 2. During exercise, RV-EDV was increased to 218 +/- 75 ml in group 1 (P less than 0.01) and to 246 +/- 79 ml in group 2. Right ventricular end-systolic volumes did not change significantly during exercise. In group 1, the left ventricular (LV) volumes were comparable to the normal; in group 2, LV-EDV was diminished at rest and during the stress test.(ABSTRACT TRUNCATED AT 250 WORDS)

Corticosteroids and ß2-agonists upregulate mitogen-activated protein kinase phosphatase 1: in vitro mechanisms.

BACKGROUND AND PURPOSE: Airway remodelling is a consequence of long-term inflammation and MAPKs are key signalling molecules that drive pro-inflammatory pathways. The endogenous MAPK deactivator--MAPK phosphatase 1 (MKP-1)--is a critical negative regulator of the myriad pro-inflammatory pathways activated by MAPKs in the airway. EXPERIMENTAL APPROACH: Herein we investigated the molecular mechanisms responsible for the upregulation of MKP-1 in airway smooth muscle (ASM) by the corticosteroid dexamethasone and the ß2-agonist formoterol, added alone and in combination. KEY RESULTS: MKP-1 is a corticosteroid-inducible gene whose expression is enhanced by long-acting ß2-agonists in an additive manner. Formoterol induced MKP-1 expression via the ß2-adrenoceptor and we provide the first direct evidence (utilizing overexpression of PKIα, a highly selective PKA inhibitor) to show that PKA mediates ß2-agonist-induced MKP-1 upregulation. Dexamethasone activated MKP-1 transcription in ASM cells via a cis-acting corticosteroid-responsive region located between -1380 and -1266 bp of the MKP-1 promoter. While the 3'-untranslated region of MKP-1 contains adenylate + uridylate elements responsible for regulation at the post-transcriptional level, actinomycin D chase experiments revealed that there was no increase in MKP-1 mRNA stability in the presence of dexamethasone, formoterol, alone or in combination. Rather, there was an additive effect of the asthma therapeutics on MKP-1 transcription. CONCLUSIONS AND IMPLICATIONS: Taken together, these studies allow us a greater understanding of the molecular basis of MKP-1 regulation by corticosteroids and ß2-agonists and this new knowledge may lead to elucidation of optimized corticosteroid-sparing therapies in the future.

Dark current measurements on a superconducting cavity using a cryogenic current comparator.

This paper presents nondestructive dark current measurements of tera electron volt energy superconducting linear accelerator cavities. The measurements were carried out in an extremely noisy accelerator environment using a low temperature dc superconducting quantum interference device based cryogenic current comparator. The overall current sensitivity under these rough conditions was measured to be 0.2 nA/Hz(1/2), which enables the detection of dark currents of 5 nA.

Enhanced characterization of serum autoantibody reactivity following HSP 60 immunization in a rat model of experimental autoimmune glaucoma.

PURPOSE: Antibodies against heat shock proteins have been identified in sera of human glaucoma patients in several studies and immunization with heat shock protein 60 (HSP 60) causes retinal ganglion cell (RGC) loss in an animal model of experimental autoimmune glaucoma. The aim of this study was to observe the time course of increased anti-retina antibody appearance in the serum and characterize the identification of prominent autoantibodies that accompany HSP 60 immunization in a rat model of experimental autoimmune glaucoma. METHODS: Eight weeks after immunization with HSP 60 retinal flatmounts were prepared and RGCs were counted in eight predefined areas and compared to controls. Serum collected before, as well as four and eight weeks after, immunization was used to detect antibody patterns against bovine retinal antigens using Western blotting techniques. These patterns were analyzed by multivariate statistical methods. Autoantibodies that were prominently increased were further identified through mass spectrometry. Intraocular pressure was measured throughout the study. RESULTS: After eight weeks, animals immunized with HSP 60 showed significant RGC loss of retinal flatmounts (P = 0.02), which was intraocular pressure independent. Early changes in antibody profiles, many of them significant upregulations, were detected. Antigens with significantly upregulated antibody reactivity after four weeks were identified as histone H2B type 1, S-arrestin, glial fibrillary acidic protein, vimentin, and heat shock protein 60. These upregulated autoantibodies returned to normal levels four weeks following their initial upregulation. Antibodies against retinaldehyde binding protein 1 on the other hand became upregulated after eight weeks. CONCLUSION: This is the first study to identify the appearance and disappearance of retinal autoantibodies in the serum of rats at several time-points following their initial upregulation in response to HSP 60 immunization in a model of experimental autoimmune glaucoma.