Cardiac magnetic field map topology quantified by Kullback-Leibler entropy identifies patients with hypertrophic cardiomyopathy.

Hypertrophic cardiomyopathy (HCM) is a common primary inherited cardiac muscle disorder, defined clinically by the presence of unexplained left ventricular hypertrophy. The detection of affected patients remains challenging. Genetic testing is limited because only in 50%-60% of all HCM diagnoses an underlying mutation can be found. Furthermore, the disease has a varied clinical course and outcome, with many patients having little or no discernible cardiovascular symptoms, whereas others develop profound exercise limitation and recurrent arrhythmias or sudden cardiac death. Therefore prospective screening of HCM family members is strongly recommended. According to the current guidelines this includes serial echocardiographic and electrocardiographic examinations. In this study we investigated the capability of cardiac magnetic field mapping (CMFM) to detect patients suffering from HCM. We introduce for the first time a combined diagnostic approach based on map topology quantification using Kullback-Leibler (KL) entropy and regional magnetic field strength parameters. The cardiac magnetic field was recorded over the anterior chest wall using a multichannel-LT-SQUID system. CMFM was calculated based on a regular 36 point grid. We analyzed CMFM in patients with confirmed diagnosis of HCM (HCM, n=33, 43.8+/-13 years, 13 women, 20 men), a control group of healthy subjects (NORMAL, n=57, 39.6+/-8.9 years; 22 women and 35 men), and patients with confirmed cardiac hypertrophy due to arterial hypertension (HYP, n=42, 49.7+/-7.9 years, 15 women and 27 men). A subgroup analysis was performed between HCM patients suffering from the obstructive (HOCM, n=19) and nonobstructive (HNCM, n=14) form of the disease. KL entropy based map topology quantification alone identified HCM patients with a sensitivity of 78.8% and specificity of 86.9% (overall classification rate 84.8%). The combination of the KL parameters with a regional field strength parameter improved the overall classification rate to 87.9% (sensitivity: 84.8%, specificity: 88.9%, area under ROC curve: 0.94). KL measures applied to discriminate between HOCM and HNCM patients showed a correct classification of 78.8%. The combination of one KL and one regional parameter again improved the overall classification rate to 97%. A preliminary prospective analysis in two HCM families showed the feasibility of this diagnostic approach with a correct diagnosis of all 22 screened family members (1 HOCM, 4 HNCM, 17 normal). In conclusion, Cardiac Magnetic Field Mapping including KL entropy based topology quantifications is a suitable tool for HCM screening.

Heterogeneous nuclear ribonucleoprotein E1B-AP5 is methylated in its Arg-Gly-Gly (RGG) box and interacts with human arginine methyltransferase HRMT1L1.

The heterogeneous nuclear ribonucleoprotein (hnRNP) family includes predominantly nuclear proteins acting at different stages of mRNA metabolism. A characteristic feature of hnRNPs is to undergo post-translational asymmetric arginine methylation catalysed by different type 1 protein arginine methyltransferases (PRMTs). A novel mammalian hnRNP, E1B-AP5, recently identified by its interaction with adenovirus early protein E1B-55 kDa, has been proposed to have a regulatory role in adenoviral and host-cell mRNA processing/nuclear export [Gabler, Schutt, Groitl, Wolf, Shenk and Dobner (1998) J. Virol. 72, 7960-7971]. Here we report that E1B-AP5 is methylated in vivo in its Arg-Gly-Gly (RGG)-box domain, known to mediate protein-RNA interactions. The activity responsible for E1B-AP5 methylation forms a complex with E1B-AP5 in vivo. The predominant mammalian arginine methyltransferase HRMT1L2 (hPRMT1) did not detectably methylate endogenous E1B-AP5 despite efficiently methylating a recombinant RGG-box domain of E1B-AP5. Using yeast two-hybrid screening we identified HRMT1L1 (PRMT2) as one of the proteins interacting with E1B-AP5. By in situ immunofluorescence we demonstrated that E1B-AP5 co-localizes with the nuclear fraction of HRMT1L1. The Src homology 3 (SH3) domain of HRMT1L1 was essential for its interaction with E1B-AP5 in vivo. We suggest that HRMT1L1 is responsible for specific E1B-AP5 methylation in vivo.

Disseminated Neocosmospora vasinfecta infection in a patient with acute nonlymphocytic leukemia.

We report Neocosmospora vasinfecta infection following chemotherapy for acute nonlymphocytic leukemia. N. vasinfecta, a plant pathogen, was identified by culture and genetic sequencing. Susceptibility testing revealed in vitro resistance for common antifungals.

Spatial features in body surface potential maps of patients with ventricular tachyarrhythmias with or without coronary artery disease.

Body surface potential maps (BSPM) from patients with coronary artery disease or no structural heart disease were analyzed with respect to their spatial features and QT/QTc dispersion in order to determine whether BSPM allows identification of patients with ventricular fibrillation. QRST integral maps and QT/QTc dispersion were acquired from simultaneous recordings of 62 ECG leads during sinus rhythm in patients with idiopathic ventricular fibrillation (n=13), ventricular fibrillation and coronary artery disease (n=22), coronary artery disease without ventricular fibrillation (n=21) and healthy controls (n=18). The Karhunen-Loeve transformation was applied to reduce the dimensionality of the data matrix of the QRST map to eight coefficients. Linear discriminant analysis allowed discrimination between idiopathic ventricular fibrillation patients and controls with high sensitivity (85%) and specificity (89%). However, discrimination between coronary artery disease patients with or without ventricular fibrillation was poor (68% and 67%, respectively). QTc dispersion calculated from BSPM was longer in idiopathic ventricular fibrillation patients than in controls (99+/-30 ms vs 70+/-14 ms, P=0.009) in contrast to QTc dispersion taken from 12-lead ECG (53+/-21 ms vs. 47+/-12 ms, P=n.s.). No significant difference was noted for coronary artery disease patients with or without ventricular fibrillation. In conclusion, repolarization disturbances detected by BSPM allow identification of ventricular fibrillation patients without structural heart disease. However, our results do not suggest a major impact of QT/QTc dispersion or QRST integral mapping for identification of ventricular fibrillation patients with coronary artery disease.

E1B 55-kilodalton-associated protein: a cellular protein with RNA-binding activity implicated in nucleocytoplasmic transport of adenovirus and cellular mRNAs.

The adenovirus type 5 (Ad5) early 1B 55-kDa protein (E1B-55kDa) is a multifunctional phosphoprotein that regulates viral DNA replication and nucleocytoplasmic RNA transport in lytically infected cells. In addition, E1B-55kDa provides functions required for complete oncogenic transformation of rodent cells in cooperation with the E1A proteins. Using the far-Western technique, we have isolated human genes encoding E1B-55kDa-associated proteins (E1B-APs). The E1B-AP5 gene encodes a novel nuclear RNA-binding protein of the heterogeneous nuclear ribonucleoprotein (hnRNP) family that is highly related to hnRNP-U/SAF-A. Immunoprecipitation experiments indicate that two distinct segments in the 55-kDa polypeptide which partly overlap regions responsible for p53 binding are required for complex formation with E1B-AP5 in Ad-infected cells and that this protein interaction is modulated by the adenovirus E4orf6 protein. Expression of E1B-AP5 efficiently interferes with Ad5 E1A/E1B-mediated transformation of primary rat cells. Furthermore, stable expression of E1B-AP5 in Ad-infected cells overcomes the E1B-dependent inhibition of cytoplasmic host mRNA accumulation. These data suggest that E1B-AP5 might play a role in RNA transport and that this function is modulated by E1B-55kDa in Ad-infected cells.

Solar absorption measurements of stratospheric OH in the UV with a Fourier-transform spectrometer.

We performed solar absorption measurements of OH in the UV using a Fourier-transform spectrometer (FTS). The experiments were carried out in the high Arctic at Ny- Alesund (79 degrees N, 12 degrees E) during the summer of 1996. We accomplished the analysis in two ways: (1) by studying single solar-absorption spectra recorded in the middle of the solar disk and (2) by utilizing the Doppler shift of two spectra, recorded on the east and west sides of the solar disk. The results of both analysis methods agree and give total columns of approximately 6 x 1013 molecules cm-2 for solar zenith angles of 60 degrees . To find out the main noise contribution in the spectra, we compared the measured and calculated signal-to-noise ratios (SNR 's). During clear-sky conditions the photon noise determines the total SNR. However, because a FTS is extremely sensitive to source fluctuations, conditions that were already slightly cloudy increased the scintillation noise, preventing OH analysis. The noise contribution caused by the instrumental sampling process itself was found to be negligible; even through two sampling positions had to be interpolated between the laser zero crossings.

Structural analysis of the adenovirus type 5 E1B 55-kilodalton-E4orf6 protein complex.

The adenovirus type 5 (Ad5) early 1B (E1B) 55-kDa (E1B-55kDa)-E4orf6 protein complex has been implicated in the selective modulation of nucleocytoplasmic mRNA transport at late times after infection. Using a combined immunoprecipitation-immunoblotting assay, we mapped the domains in E1B-55kDa required for the interaction with the E4orf6 protein in lytically infected A549 cells. Several domains in the 496-residue 55-kDa polypeptide contributed to a stable association with the E4orf6 protein in E1B mutant virus-infected cells. Linker insertion mutations at amino acids 180 and 224 caused reduced binding of the E4orf6 protein, whereas linker insertion mutations at amino acid 143 and in the central domain of E1B-55kDa eliminated the binding of the E4orf6 protein. Earlier work showing that the central domain of E1B-55kDa is required for binding to p53 and the recent observation that the E4orf6 protein also interacts with the tumor suppressor protein led us to suspect that p53 might play a role in the E1B-E4 protein interaction. However, coimmunoprecipitation assays with extracts prepared from infected p53-negative H1299 cells established that p53 is not needed for the E1B-E4 protein interaction in adenovirus-infected cells. Using two different protein-protein interaction assays, we also mapped the region in the E4orf6 protein required for E1B-55kDa interaction to the amino-terminal 55 amino acid residues. Interestingly, both binding assays established that the same region in the E4orf6/7 protein can potentially interact with E1B-55kDa. Our results demonstrate that two distinct segments in the 55-kDa protein encoding the transformation and late lytic functions independently interact with p53 and the E4orf6 protein in vivo and provide further insight by which the multifunctional 55-kDa EIB protein can exert its multiple activities in lytically infected cells and in adenovirus transformation.

[Analysis of QRST integral and QT dispersion by body surface potential mapping in patients with malignant ventricular arrhythmias]. / Analyse von QRST-Integral und QT-Dispersion mittels "Body surface potential mapping" bei Patienten mit malignen ventrikulären Arrhythmien.

In a retrospective analysis in 74 patients with coronary artery disease or no obvious heart disease, the value of "body surface potential mapping" for the identification of repolarization abnormalities was investigated compared to the standard 12-lead ECG. In patients with idiopathic ventricular fibrillation the number of extrema in the QRST integral map was significantly higher than in the control group (3.15+/-0.99 vs. 2.17+/-0.51, p<0.001) and the QT dispersion was also higher (0.10+/-0.03 vs. 0.07+/-0.01, p<0.001), whereas there was no difference between either group in the 12-lead ECG QT dispersion. In patients with coronary artery disease the number of extrema in the QRST integral map and QT dispersion were also higher compared to the control group, but there were no significant differences between patients with or without aborted sudden cardiac death.In conclusion, BSPM identifies repolarization abnormalities not detected by 12-lead ECG, thereby identifying a potential reason for cardiac arrest in patients without overt heart disease. The usefulness of this technique for risk stratification in patients with coronary artery disease remains to be elucidated.

The mode of onset of ventricular tachycardia. A patient-specific phenomenon.

AIMS: The purpose of our study was to investigate the electrical trigger modes of monomorphic ventricular tachycardia, by analysing stored intracardiac electrograms, and to identify haemodynamic or electrocardiographic predictors in patients with cardioverter-defribrillators. METHODS: We recorded 286 episodes of monomorphic ventricular tachycardia in 38 patients with at least three events. The electrical triggers were characterized by the morphology number, complexity, and coupling interval of premature depolarizations preceding the ventricular tachycardia. We also evaluated clinical and electrocardiographic data. RESULTS: We found two basic electrical trigger modes. Two hundred and sixteen events (75%) were observed to have no RR-interval variations before onset, while 70 episodes (25%) had a short-long-short sequence. These episodes invariably featured increased QT disperson. In 31 of 38 patients (82%), the ventricular tachycardias were always initiated by the same mode of onset. In eight patients, the monomorphic ventricular tachycardias were always triggered by short-long-short sequences. In seven patients, more than one onset mechanism was observed. CONCLUSIONS: Two basic modes of onset were responsible for monomorphic ventricular tachycardia: one without RR-interval variations immediately prior to onset and another characterized by short-long-short sequences and increased QT dispersion. The mechanisms were largely patient-specific and not related to cardiac diagnosis or left ventricular function.

[Not Available]. / Sprachschichten der alchemie.

The language of alchemy may be divided into five layers. - Within the first layer the alchemists explicitly and undisguisedly conveyed chemical informations. But they described chemical substances and chemical operations in a way which reminds us of sociological or psychological descriptions and the treatment of complex personalities. - Within the second layer the alchemists also gave chemical informations albeit encoded in much the same way as craftmen's guilds encoded their knowledge to fend off competition from outsiders. - Within the third layer the alchemists refer to verbal associations and to myths from their cultural environment so that their texts can only be properly understood within a certain cultural context. As the language of this layer was often handed down to later generations of alchemists who were not acquainted with the specific cultural context it tended to cause confusions and misunderstandings. - Within the fourth layer the alchemists deliberately used an arcane language for within this layer they tried to convey the paradoxa of alchemy. - Within the fifth ayer alchemy is treated as a literary tradition in which all classical texts contain the truth but no text does it in a fully comprehensible way. - These last two layers show that au fond alchemy is not a process of riddle solving but a quest for cosmic secrets. This gives us a demarcation criterion to distinguish alchemy from genuine natural sciences.

[Diagnosis of hepatitis C virus (HCV) infection: diagnostic value of the anti-HCV test]. / Diagnose der Hepatitis-C-Virus(HCV)-Infektion: Diagnostische Wertigkeit des Anti-HCV-Tests.

Different groups of patients were analysed for antibody to hepatitis-C-virus (anti-HCV). A high prevalence was found in individuals with parenteral exposure (chronic Non-A, Non-B hepatitis, 77.5%, drug addicts 84.5%), while blood donors had a prevalence of 0.51%, this was significantly higher in patients with chronic type B hepatitis (30%), in homosexuals (22.5%) and in patients with different types of autoimmune hepatitis (57.2%). This indicates that differential diagnosis of chronic hepatitis and indications for alpha interferon therapy is not possible by simply anti-HCV testing. Further studies are required to establish the diagnostic value of the anti-HCV test.

Radiosynthesis of [14C]acarbose.

Acarbose (O-4,6-dideoxy-4-[[(1S, 4R, 5S, 6S)-4,5,6-trihydroxy-3- (hydroxymethyl)-2-cyclohexen-1-yl]amino]-a-D-glucopyranosyl-(1---- 4)-O-a-D- glucopyranosyl-(1----4)-4-glucopyranose, Bay g 5421), an a-glucosidase inhibitor from Actinoplanes, has been developed for the treatment of diabetes mellitus. To investigate the pharmacokinetics and the biotransformation, 14C-labelled acarbose ([14C]Bay g 5421) was required. About 37 GBq (1 Ci) D-[U-14C]glucose was used as a precursor to obtain [14C]acarbose with a radiochemical yield of between 1.58 and 2.56%. For fermentation purposes resting cells of the Actinoplanes mutant SN 1667/47 were used under cometabolism conditions with a 10-fold excess of maltose. The specific radioactivities achieved in individual preparations were 7.77 MBq/mg (210 microCi/mg), 8.03 MBq/mg (217 microCi/mg), and 9.14 MBq/mg (247 microCi/mg), with a radiochemical purity of greater than 98% in each case. By hydrolysis and subsequent investigation of the hydrolysis products it was shown that [14C]carbon atoms originating from the radioactive glucose are present only in the core and not in the maltose unit of [14C]acarbose.

Fate and effects of the alpha-glucosidase inhibitor acarbose in humans. An intestinal slow-marker perfusion study.

The alpha-glucosidase inhibitor acarbose has been successfully used in diabetic patients to decrease the postprandial rise in blood glucose. The aim of the present experiments was to investigate the fate and effects of acarbose along the small intestine using a slow-marker perfusion technique. In 8 healthy volunteers, jejunal and ileal loads of acarbose, glucose, and total carbohydrates were determined following a liquid, 400-kcal formula meal containing either 200 mg of acarbose or placebo. Preprandial and postprandial plasma concentrations of glucose and several polypeptide hormones were determined. Recovery of acarbose during 4 h was 65% +/- 9% (mean +/- SEM) of ingested dose in the ileum but 94% +/- 9% in the jejunum, indicating that the compound was neither degraded nor absorbed by the intestine to a major degree. After acarbose administration, ileal loads of glucose and total carbohydrates were considerably higher, whereas postprandial plasma concentrations of glucose, insulin, and gastric inhibitory polypeptide were lower when compared with placebo. The retardation of carbohydrate digestion to be inferred from these findings is confirmed by significantly elevated plasma concentrations of enteroglucagon after acarbose administration compared with placebo administration.

Pyroglutamyl-aprotinin, a new aprotinin homologue from bovine lungs--isolation, properties, sequence analysis and characterization using 1H nuclear magnetic resonance in solution.

A new Kunitz-inhibitor, which is different from aprotinin was extracted from bovine lungs with methanol, further purified by affinity chromatography on trypsin-Sepharose CL-6B and by repeated cation exchange chromatography on CM-Sephadex C-25. The inhibitor, which is less basic than aprotinin was characterized by polyacrylamide gel electrophoresis and ion-exchange HPLC. The N-terminus is blocked by pyroglutamic acid (Glu-1). After enzymatic removal of this residue with pyroglutamate aminopeptidase, complete identity with the primary structure of aprotinin was established by sequencing the inhibitor, which had been oxidized with performic acid, and by sequencing a tryptic fragment. The occurrence of the inhibitor, which can be denoted as pyroglutamyl-aprotinin or Glu-1-aprotinin, but which cannot be distinguished from aprotinin regarding its inhibitory specificity, is obviously the result of a different proteolytic processing of the bovine aprotinin precursor. By using CD and NMR-techniques it was shown that the N-terminus of the inhibitor is blocked, and that the conformation and the internal mobility correspond with those of aprotinin.