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Treating therapy-resistant patients with inherited arrhythmia syndromes can be difficult and left cardiac sympathetic denervation (LCSD) might be a viable alternative treatment option. We provide an overview of the indications and outcomes of LCSD in patients with inherited arrhythmia syndromes in the only tertiary referral centre in the Netherlands where LCSD is conducted in these patients. This was a retrospective study, including all patients with inherited arrhythmia syndromes who underwent LCSD in our institution between 2005 and 2013. LCSD involved ablation of the lower part of the left stellate ganglion and the first four thoracic ganglia. Seventeen patients, 12 long-QT syndrome (LQTS) patients (71 %) and 5 catecholaminergic polymorphic ventricular tachycardia (CPVT) patients (29 %), underwent LCSD. Most patients (94 %) were referred because of therapy-refractory cardiac events. In 87 % the annual cardiac event rate decreased. However, after 2 years the probability of complete cardiac event-free survival was 59 % in LQTS and 60 % in CPVT patients. Two patients (12 %) had major non-reversible LCSD-related complications: one patient suffered from a Harlequin face post-procedure and one severely affected LQT8 patient died the day after LCSD due to complications secondary to an arrhythmic storm during the procedure. LSCD for inherited arrhythmia syndromes, which is applied on a relatively small scale in the Netherlands, reduced the cardiac event rate in 87 % of the high-risk patients who had therapy-refractory cardiac events, while the rate of major complications was low. Therefore, LSCD seems a viable treatment for patients with inherited arrhythmia syndromes without other options for therapy.

Communication between the aortic sinus and a cardiac chamber is a rare anomaly that can be diagnosed in children and young adults. We describe two cases with a tunnel between the aortic sinus and right atrium, discuss diagnostic modalities, and review the literature on this anomaly.Communication between the aortic sinus and a cardiac chamber is a rare anomaly that can be diagnosed in children and young adults. We describe two cases with a tunnel between the aortic sinus and right atrium, discuss diagnostic modalities, and review the literature on this anomaly.

20 % of patients with a systemic RV are pacemaker dependent, and unsuitable to undergo cardiac magnetic resonance (CMR). Multidetector row computed tomography (MDCT) could provide a reproducible alternative to CMR in these patients. The aim of this study was to compare variability of MDCT with CMR.Thirty-five patients with systemic RV underwent either MDCT (n?=?15) or CMR (n?=?20). Systemic RV volumes and ejection fraction were obtained, and intra- and interobserver variability for both modalities were assessed and compared.We found the intra- and interobserver variability of volumes and function measurements of the systemic RV obtained with MDCT to be higher compared with those obtained with CMR. However, these differences in variability were not significant, the only exception being the interobserver variability of systemic RV stroke volume.MDCT provides a reproducible alternative to CMR for volumes and function assessment in patients with a systemic RV.

Abstract: Aortic coarctation is a common congenital cardiac defect, which can be diagnosed over a wide range of ages and with varying degrees of severity. We present two cases of patients diagnosed with aortic coarctation in adulthood. Both patients were treated by an endovascular approach. These cases demonstrate the variety of indications in which percutaneous treatment is an excellent alternative for surgical treatment in adult native coarctation patients.

Protein phosphorylation plays a key role in cell regulation and identification of phosphorylation sites is important for understanding their functional significance. Here, we present an artificial neural network algorithm:  NetPhosK (http://www.cbs.dtu.dk/services/NetPhosK/) that predicts protein kinase A (PKA) phosphorylation sites. The neural network was trained with a positive set of 258 experimentally verified PKA phosphorylation sites. The predictions by NetPhosK were validated using four novel PKA substrates:  Necdin, RFX5, En-2, and Wee 1. The four proteins were phosphorylated by PKA in vitro and 13 PKA phosphorylation sites were identified by mass spectrometry. NetPhosK was 100% sensitive and 41% specific in predicting PKA sites in the four proteins. These results demonstrate the potential of using integrated computational and experimental methods for detailed investigations of the phosphoproteome. Keywords: protein kinase A • phosphorylation site prediction • neural network analysis • mass spectrometry

Myocardial perfusion imaging with technetium99mlabelled methoxyisobutyl isonitrile single photon emission computed tomography 99mTcMIBI SPECT has proven to be an important clinical procedure in assessing the severity of myocardial ischaemia. The uptake and clearance of 99mTcMIBI by the myocardium is affected by cell viability and membrane integrity. Consequently, infectious diseases, such as myocarditis, may also affect myocardial perfusion by inducing local inflammation and necrosis. We compared 99mTcMIBI myocardial perfusion imaging with other heart monitoring methods in order to assess its value in the diagnosis of children with Coxsackie viral myocarditis. We examined 46 patients age, 3–12 years with Coxsackie viral myocarditis using 99mTcMIBI myocardial perfusion imaging and compared the perfusion data with myocardial enzymes, electrocardiographic findings and echocardiography. Regions of hypoperfusion were found in all 46 patients. Seventeen patients 37 showed two or more areas of diminished perfusion. Myocardial hypoperfusion was mildtomoderate <30 in 33 72 patients and severe >30 in 13 28 patients. Characteristic creatinekinase isoenzyme CKMB increases, STT segment changes and diminished heart function were significantly correlated with reduced myocardial perfusion all comparisons P<0.05. The results of this study suggest that the presence of myocardial uptake of 99mTcMIBI may be a marker of myocardial inflammation and necrosis. All 46 patients with Coxsackie viral myocarditis showed a certain degree of reduced perfusion. When the perfusion findings were compared with other parameters, it was shown that myocardial enzyme levels, STT segment changes and left ventricular function correlated well with the 99mTcMIBIestablished perfusion defect severity. 99mTcMIBI SPECT imaging is therefore helpful in providing additional diagnostic information in patients with Coxsackie viral myocarditis.

Objective:The cytomegalovirus (CMV) antigenemia consists of the detection of CMV pp65 in the nucleus of polymorphonuclear granulocytes (PMN), but it is unclear where and how PMN pick up virus particles or proteins. In an in vitro model for CMV antigenemia we investigated the mechanism of pp65 uptake by PMN that results in its expression in the nucleus. Methods:A series of inhibitors of different mechanisms was used to study the uptake of pp65 by PMN during coculture with CMV-infected endothelial cells and we performed a morphological analysis by light and transmission electron microscopy. Results: Nocodazole and cytochalasin B inhibited uptake of pp65 by PMN with 59.4 ± 14.1 and 73.3 ± 12.7%, respectively. The presence of anti-CMV hyperimmune globulin or lactoferrin during coculture reduced the number of pp65-positive PMN with 45.8 ± 7.0 or 40.6 ± 3.2%. Furthermore, a small number of the pp65-positive PMN obtained after coculture had fused to large cells with multilobed nuclei. PMN were observed that enclosed viral particles as well as free viral particles containing PMN in the cytoplasm. Conclusion:Fusion of viral particles with PMN and phagocytosis are both involved in the uptake of pp65.

Electrophysiological and new immunohistochemical markers have resulted in a better understanding of the development of the cardiac conduction system. In the primary heart tube the blood is pumped forward by peristaltic contractions as a result of alternating slow and fast conducting segments. At later stages a more specialized conduction system develops. The contribution of the primary fold and the sinus venosus to the formation of the cardiac conduction system is discussed as well as the role of extracardiac cells such as neural crest and epicardial derived cells. These new developmental aspects of the cardiac conduction system could provide an explanation of the position conduction system in heart malformations and certain arrhythmias in patients with structurally normal hearts.

The apoAI-CIII-AIV gene cluster, located on chromosome 11, contributes to the phenotype of familial combined hyperlipidemia (FCH), but this contribution is genetically complex. Combinations of haplotypes, based on three restriction enzyme polymorphisms: XmnI and MspI sites, 5' of the start site of the apoA-I gene and SstI polymorphism in the 3' untranslated region of exon 4 of the apoC-III gene, were analyzed to characterize their effect on the expression of severe hyperlipidemia. An epistatic interaction was demonstrated: the S2 allele on one haplotype was synergistic in its hyperlipidemic effect to the X2M2 allele on the other haplotype (Dallinga-Thie, G. M. et al. J. Clin. Invest. 1997. 99: 953-961). In the present study two additional polymorphic sites in the insulin response element (IRE) of the apoC-III gene promoter, T-455C: FokI restriction site, C-482T: MspI restriction site, were studied in 34 FCH pedigrees including 34 probands, 220 hyperlipidemic relatives, 300 normolipidemic relatives, and 236 spouses. In contrast to the earlier data for the other polymorphisms in this gene cluster (XmnI, MspI/AI, and SstI), there were no differences in frequency distributions of the T-455C and the C-482T variants between probands, hyperlipidemic and normolipidemic relatives and spouses. No significant associations between plasma lipid traits and DNA variants in the IRE were observed. Analysis of combinations of haplotypes based on the five polymorphisms in the gene cluster provided further evidence for a dominant role of the SstI polymorphism as a major susceptibility locus in FCH. The inclusion of the IRE markers did not improve genetic informativeness, nor our understanding of the observed synergistic relationship associated with the high risk combination of haplotypes in FCH families.

We have studied the morphology and cytochemistry in relation to the immunological phenotyping and functional properties of T cells from eight patients with chronic T? lymphocytosis. At the light microscopic level the morphology of the patients' lymphocytes was similar to that described for large granular lymphocytes. Ultrastructurally, a division into two groups could be made on differences in the amount of cytoplasm and the location and the more irregular form of the nuclei. The lymphocytes of one group (five patients) had in common the phenotype Fc?+, T3+, T4-, T8+, Ia-, M1- and demonstrated (with the exception of one patient) the same functions: presence of K-cell activity, absence of NK, helper and suppressor cell activities. In the other group (three patients), the lymphocytes of one patient showed the same phenotype and functions as those indicated above. The other two patients both lacked the T8 antigen on their lymphocytes but were different with regard to other surface markers. In addition, their cells were functionally identical: both demonstrated NK- and K-cell activity. Thus in this group of eight patients with chronic T? lymphocytosis, the immunological and functional subdivision paralleled in part a morphological division at the ultrastructural level.

A double immunogold-labeling method in immunoelectron microscopy was used for simultaneous detection of two antigens by monoclonal antibodies [OKT 8 (CD 8), anti-Leu-7, anti-Leu-11b (CD 16)] on lymphocytes in suspension. The combination of gold probe size (5 nm and 15 nm) and monoclonal antibody was found to be decisive for detecting double-labeled cells with the OKT 8+, Leu-11b+ phenotype. The combinations of OKT 8 labeled with the 5-nm gold probe (OKT 8(5] and anti-Leu-11b with the 15-nm gold probe (Leu-11b15) gave double-labeled cells; the reverse situation, using OKT 8 with a 15-nm gold probe (OKT 8(15] and anti-Leu-11b with a 5-nm gold probe (Leu-11b5), did not. Double-labeled OKT 8+, Leu-7+ cells were detected irrespective of which gold probe combination was applied. Our findings indicate that although the double immunogold-labeling method is well suited for study of lymphocyte subsets, it is important to determine suitable combinations of gold probe sizes and monoclonal antibodies for the lymphocyte subset under study, taking into account surface antigen density, so that double labeling ensues.

Summary Platelet concentrates were pretreated with a stable synthetic prostacyclin analogue (Iloprost) at two different concentrations before the second centrifugation step (pelleting step) of preparation. This resulted in loss of platelet sensitivity to aggregating agents. To mimic the situation after transfusion and to assess the reversibility of platelet inhibition, platelets were washed during and after storage and resuspended in fresh-frozen autologous plasma. The Iloprost-treated and washed platelets exhibited an increased sensitivity to the aggregating agents, compared with the control platelets (p< 0.01). Post-storage recovery of the synergistic aggregation was more than 80% of prestorage aggregation, ?-thromboglobulin (?TG) release and thromboxane B2 (TXB2) formation were significantly inhibited in Iloprost-treated platelets (p< 0.01). After the second centrifugation step, ?TG release was 0.7%±0.3%, compared with 2.7%±0.9% for the controls. TXB2 was 99±91 pg/ml, compared with 495±356 pg/ml for the controls. Platelet morphology and ultrastructure were well preserved during 5-day storage. In addition, Iloprost exerted a cytoprotective effect, as evidenced by the significant reduction in lactate dehydrogenase leakage. Post-storage LDH was 378±159 and 415±239 U/l respectively by the two Iloprost concentrations, compared with 1180±937 U/l for the control platelets. The inhibitory and cytoprotective effects of Iloprost were sustained throughout storage, in contrast to the effect of PGE1 (Prostin) which was limited to the early phase of storage.

Normal donor peripheral blood mononuclear cells incubated for 24 h with sera from patients with Hodgkin's disease were investigated by electron microscopy for the presence of parallel tubular structures (PTS) and/or amorphous electrondense granules (large granular lymphocytes = LGL). In comparison with normal human serum, 14 out of 29 sera of the patients induced a marked increase in the percentage of LGL. From a limited number of experiments it was likely that this increase is paralleled by an increase in Fc? receptor-bearing cells after the incubation. This serum effect did not show a correlation with the number of Fc? receptor-positive lymphocytes in the peripheral blood of the patients.

Peripheral blood lymphocytes were fractionated in T, B and Null cell enriched subsets by means of sheep red blood cell rosette (ESRBC) sedimentation and nylon wool adherence. The ultrastructural features of these subpopulations were investigated.

PhosphoBase is a database of experimentally verified phosphorylation sites. Version 1.0 contains 156 entries and 398 experimentally determined phosphorylation sites. Entries are compiled and revised from the literature and from major protein sequence databases such as SwissProt and PIR. The entries provide information about the phosphoprotein and the exact position of its phosphorylation sites. Furthermore, part of the entries contain information about kinetic data obtained from enzyme assays on specific peptides. To illustrate the use of data extracted from PhosphoBase we present a sequence logo displaying the overall conservation of positions around serines phosphorylated by protein kinase A (PKA). PhosphoBase is available on the WWW at http://www.cbs.dtu.dk/databases/PhosphoBase/

Several DNAs representing the genome of the avian acute leukemia virus OK 10 were isolated by molecular cloning from a transformed quail cell line, 9C, which contained at least six OK 10 proviruses. Recombinant lambda phages harboring the OK 10 genome and additional flanking cellular DNA sequences were studied by restriction endonuclease mapping and hybridization to viral cDNA probes. Six of the clones represented complete proviruses with similar, if not identical, viral sequences integrated at different positions in the host DNA. The organization of the OK 10 genome was determined by electron-microscopic analysis of heteroduplexes formed between the cloned OK 10 DNA and DNAs representing the c-myc gene and the genomes of two other avian retroviruses, Rous-associated virus-1 and MC29. The results indicated that the OK 10 proviral DNA is about 7.5 kilobases in size with the following structure: 5'-LTR-gag-delta polmyc-delta env-LTR-3', where LTR indicates a long terminal repeat. The oncogene of OK 10, v-mycOK 10, forms a continuous DNA segment of around 1.7 kilobases between pol and env. It is similar in structure and length to the v-myc gene of MC29, as demonstrated by restriction endonuclease and heteroduplex analyses. Two of the OK 10 proviruses were tested in transfection experiments: both DNAs gave rise to virus with the transforming capacities of OK 10 when Rous-associated virus-1 was used to provide helper virus functions.

The immunological efficacy and the absence of communicability of live attenuated measles-virus vaccine in children has been described by Katz and Enders1 and by others2-10 Since immunity to natural measles infection shortly after vaccination has also been reported,3,4 the final acceptance of this vaccine for routine pediatric immunization will presumably depend on the persistence of the immunity conferred and on the severity of reactions to the vaccination.The main purpose of the study undertaken here has been to gain further information regarding the reactions caused by a subcutaneously inoculated attenuated measles-virus vaccine prepared from the chick-embryo-adapted Edmonston strain in institutionalized children and the effect of γ-globulin on such reactions. In addition, the serological response to vaccination was tested by complement fixation (CF) and hemagglutination inhibition (HI) techniques. The full report of this study will be published elsewhere.11 MATERIAL AND METHODS Design and Operation of Study.—The

Summary Sera from 716 inhabitants of the Aland archipelago were examined for hemagglutination-inhibiting antibodies with the diphasic tick-borne encephalitis virus. The antibodies were found in 13.1 per cent of the whole series. There was no significant difference in antibody rate between males and females. It was noted that the antibody rate of the population increased almost linearly with age, which indicates that the human population of the area is exposed to a relatively constant risk of infection. Great local variations in the incidence of the antibodies were found among the residents of neighboring islands. No accumulation by families could be established in the distribution of immunity, which speaks against transmission of the virus through milk in the area.

This study presents an improved approach to common‐conversion point stacking of converted body waves that incorporates scattering kernels, accurate and efficient measurement of stack uncertainties, and an alternative method for estimating free surface seismic velocities. To better separate waveforms into the Pand SVcomponents to calculate receiver functions, we developed an alternative method to measure near‐surface compressional and shear wave velocities from particle motions. To more accurately reflect converted phase scattering kernels in the common‐conversion point stack, we defined new weighting functions to project receiver function amplitudes only to locations where sensitivities to horizontal discontinuities are high. To better quantify stack uncertainties, we derived an expression for the standard deviation of the stack amplitude that is more efficient than bootstrapping and can be used for any problem requiring the standard deviation of a weighted average. We tested these improved methods on Spphase data from the Anatolian region, using multiple band‐pass filters to image velocity gradients of varying depth extents. Common conversion point stacks of 23,787 Spreceiver functions demonstrate that the new weighting functions produce clearer and more continuous mantle phases, compared to previous approaches. The stacks reveal a positive velocity gradient at 80–150 km depth that is consistent with the base of an asthenospheric low‐velocity layer. This feature is particularly strong in stacks of longer period data, indicating it represents a gradual velocity gradient. At shorter periods, a lithosphere‐asthenosphere boundary phase is observed at 60–90 km depth, marking the top of the low‐velocity layer. This paper presents a new method that more accurately incorporates the physics of seismic scattering into how the wave records are combined to form images of gradients in seismic velocity structure. This method was tested on data from the Anatolian region, where the asthenosphere is known to have low seismic wave velocities, consistent with high mantle temperatures and possibly small fractions of partial melt, as suggested by the presence of volcanic fields at the surface. However, the depth of the asthenospheric low‐velocity layer is not well known. In this study, we locate this low‐velocity mantle layer by applying the newly developed imaging method to seismic shear waves that convert to compressional waves at the velocity gradients that mark the layer boundaries. This study is the first to clearly resolve both the lower and upper margins of the asthenosphere for the whole region. The top of the layer corresponds to the lithosphere‐asthenosphere boundary at 60–90 km depth, and this velocity gradient is localized in depth. However, the bottom boundary, which lies at depths of 80–150 km, occurs over a broader depth range. A new approach to common conversion point stacking based on scattering kernels is developedA fast and accurate quantification of the standard deviation of the weighted stack average is derivedAn anomalously low‐velocity asthenosphere beneath Anatolia is indicated by velocity discontinuities imaged with multifrequency data

PhosphoBase contains information about phosphorylated residues in proteins and data about peptide phosphorylation by a variety of protein kinases. The data are collected from literature and compiled into a common format. The current release of PhosphoBase (October 1998, version 2.0) comprises 414 phosphoprotein entries covering 1052 phosphorylatable serine, threonine and tyrosine residues. The kinetic data from peptide phosphorylation assays for approximately 330 oligopeptides is also included. The database entries are cross-referenced to the corresponding records in the Swiss-Prot protein database and literature references are linked to MedLine records. PhosphoBase is available via the WWW at http://www.cbs.dtu. dk/databases/PhosphoBase/

An indirect immunofluorescence test for detection of serum antibodies specific for nephropathia epidemica (NE) has been developed with use of acetone-fixed cryostat sections of the lungs of bank voles (Clethrionomys glareolus) that had been trapped from the NE-endemic area in Finland as antigen. NE antigen was detected as distinct fluorescence in the cytoplasm of alveolar and macrophage-like cells. The 16 patients studied included typical cases from an endemic area, cases from a family outbreak, and cases in a laboratory staff which had had close contact with infected bank voles. Antibodies reacting with antigen in the lung sections developed in all of the patients, but they were not found in the preimmune sera of the patients, in the sera of patients with other renal diseases, or in the sera of healthy individuals, with the exception of a member of the laboratory staff who had lived in the endemic area for 20 years. No specific IgM antibodies to NE could be detected. The rise in titer of antibodies to NE was characteristically prolonged, and elevated antibody levels persisted for many years.