Long-term outcome of perimembranous VSD closure using the Nit-Occlud® Lê VSD coil system.

OBJECTIVE: This study presents data from the admission trial to show the feasibility, safety and effectiveness of the Nit-Occlud® Lê VSD in the treatment of perimembranous ventricular septal defects with an aneurysmal configuration and a diameter up to 8 mm. BACKGROUND: The majority of ventricular septal defects (VSD) are still closed surgically, while a less invasive transcatheter treatment by closure devices is available. Device-based closure is reported to be associated with the risk of complete atrio-ventricular block, especially with double-disc devices in perimembranous defects. METHODS: In six tertiary centers in Germany and Israel, an interventional closure of a periembranous VSD was attempted in 88 patients using the Nit-Occlud® Lê VSD. RESULTS: The interventional VSD closure was performed in 85 patients. Patients had a median age of 8.0 (2-65) years and a median body weight of 26.7 (10-109) kg. A complete closure of the defects was achieved in 85.4% 2 weeks after device implantation, in 88.9% after three months and in 98.6% at the 5-year follow-up. There was no incidence of death during the study nor did any patient suffer of permanent atrio-ventricular block of higher degree. Serious adverse events, by definition, are potentially life-threatening or require surgery to correct, while major serious events require medical or transcatheter intervention to correct. The study results exhibit a serious adverse event rate of 3.5% (3/85 patients) and a major adverse event rate of 5.9% (5/85 patients). CONCLUSION: The Nit-Occlud® Lê VSD coil offers the possibility of an effective and safe approach in patients with aneurysmal perimembranous ventricular septal defects.

Genotype-Phenotype Correlation in Children: The Impact of FBN1 Variants on Pediatric Marfan Care.

Currently, no reliable genotype-phenotype correlation is available for pediatric Marfan patients in everyday clinical practice. We investigated correlations of FBN1 variants with the prevalence and age of onset of Marfan manifestations in childhood and differentiated three groups: missense/in-frame, splice, and nonsense/frameshift variants. In addition, we differentiated missense variants destroying or generating a cysteine (cys-missense) and alterations not affecting cysteine. We categorized 105 FBN1-positive pediatric patients. Patients with cys-missense more frequently developed aortic dilatation (p = 0.03) requiring medication (p = 0.003), tricuspid valve prolapse (p = 0.03), and earlier onset of myopia (p = 0.02) than those with other missense variants. Missense variants correlated with a higher prevalence of ectopia lentis (p = 0.002) and earlier onset of pulmonary artery dilatation (p = 0.03) than nonsense/frameshift, and dural ectasia was more common in the latter (p = 0.005). Pectus excavatum (p = 0.007) appeared more often in patients with splice compared with missense/in-frame variants, while hernia (p = 0.04) appeared earlier in the latter. Findings on genotype-phenotype correlations in Marfan-affected children can improve interdisciplinary therapy. In patients with cys-missense variants, early medical treatment of aortic dilatation seems reasonable and early regular ophthalmologic follow-up essential. Patients with nonsense/frameshift and splice variants require early involvement of orthopedic specialists to support the growing child.

The transition of pediatric Marfan patients to adult care: a challenge and its risks.

BACKGROUND: Care for patients with Marfan syndrome (MFS) has improved substantially in recent decades. Increasing clinical knowledge and genetic analysis allow early diagnosis of the disease in childhood. Because of the earlier initiation to preventive and medical treatment, patients' life expectancy has risen. To ensure optimal care, pediatric patients require a safe follow-up regime, multidisciplinary care, and a safe transition to adult care. METHODS: We collected a sample of 149 pediatric Marfan patients, of whom 34 patients had already been transferred to adult care or who were currently transitioning. First, we evaluated clinical aspects of patients that manifest in childhood and are present in the transition process. Second, we analyzed the transition process itself. RESULTS: We found age-dependent manifestation of organ pathologies. Dilatation of the sinus of Valsalva showed a particularly high prevalence during the transition process and 62% of patients required medical treatment. Mean onset of aortic root dilatation was 9.9±5.8 years. Concerning systemic manifestation in MFS skin striae, wrist and thumb sign, and reduced elbow extension occurred significantly more often in patients who were transitioning than in younger children with MFS. All other clinical Marfan features showed an increased prevalence in patients who were transitioning compared with younger patients. In our cohort, transition was successful in 20 patients (58.9%), 12 patients (35.3%) are still in the transition process and 2 patients (5.9%) were lost to follow up. CONCLUSIONS: Marfan patients in the transition process are already under a chronic disease condition with a high onset of especially cardiovascular pathologies. Although early medical treatment in childhood is effective, the pathologies of the connective tissue require lifelong attention and influence life in many ways. The big challenge during transition is the double change of responsibility from the parents and pediatric doctor to the patient and adult doctor. Consequently, patients in transition process require special attention and close contact with the doctor and the family. A reevaluation by the supervising pediatric Marfan specialist of the successful transition to adult care is indispensable before the pediatric care of Marfan patients is completed.

[Current surgical and adjuvant therapy concepts of malignant tumors of the facial skin and the pinna]. / Aktueller Stand der chirurgischen und adjuvanten Therapie der Malignome der Gesichtshaut und der Ohrmuschel.

Malignant tumors of the skin had been a rare entity 2 decades ago. Today they are spread rapidly worldwide. Malignant neoplasms of the skin, the largest human organ, may occur from all structures and layers. While previously skin cancer -occurred mainly after the age of 60, the incidence increases now in younger ages. Strong sunburns in the childhood and before the age of 20 are important risk factors for the development of malignancies of the skin. An increased exposure to UV rays is found especially in the facial skin, where basal cell carcinoma, squamous cell carcinoma, malignant melanoma and Merkel cell carcinomas are the most common malignancies. Early diagnosis of malignancies and therapy-oriented mostly surgical approaches are crucial for the prognosis of all skin cancers. Therefore under the aspect of the increasing incidence these topics will be pointed out according to the latest findings including current multimodal therapy concepts and future treatment options.

The effect of carbon black reinforcement on the dynamic fatigue and creep of polyisobutylene-based biomaterials.

This paper investigates the structure-property relationship of a new generation of poly(styrene-b-isobutylene-b-styrene) (SIBS) block copolymers with a branched (dendritic) polyisobutylene core with poly(isobutylene-b-para-methylstyrene) end blocks (D_IBS), and their carbon black (CB) composites. These materials display thermoplastic elastomeric (TPE) properties, and are promising new biomaterials. It is shown that CB reinforced the block copolymer TPEs, effectively delayed the oxidative thermal degradation of the D_IBS materials, and greatly improved their dynamic fatigue performance. Specifically, the dynamic creep of a CB composite was comparable to that of chemically crosslinked and silica-reinforced medical grade silicone rubber, used as a benchmark biomaterial.

Investigation of structure-property relationships of polyisobutylene-based biomaterials: Morphology, thermal, quasi-static tensile and long-term dynamic fatigue behavior.

This study examines the morphology, thermal, quasi-static and long-term dynamic creep properties of one linear and three arborescent polyisobutylene-based block copolymers (L_SIBS31, D_IBS16, D_IBS27 and D_IBS33). Silicone rubber, a common biopolymer, was considered as a benchmark material for comparison. A unique hysteretic testing methodology of Stepwise Increasing Load Test (SILT) and Single Load Test (SLT) was used in this study to evaluate the long-term dynamic fatigue performance of these materials. Our experimental findings revealed that the molecular weight of polyisobutylene (PIB) and polystyrene (PS) arms [M(n)(PIB(arm)) and M(n)(PS(arm))], respectively had a profound influence on the nano-scaled phase separation, quasi-static tensile, thermal transition, and dynamic creep resistance behaviors of these PIB-based block copolymers. However, silicone rubber outperformed the PIB-based block copolymers in terms of dynamic creep properties due to its chemically crosslinked structure. This indicates a need for a material strategy to improve the dynamic fatigue and creep of this class of biopolymers to be considered as alternative to silicone rubber for biomedical devices.

The impact of shunt type on palliative outcomes in neonates and infants with diminished pulmonary blood flow.

We analyzed early and intermediate outcomes in cyanotic neonates (n = 43) and infants (n = 26) requiring palliation with either a modified Blalock-Taussig shunt (MBT) or a central aortopulmonary shunt (CAP). Between 1995 and 2009, 69 consecutive patients underwent an MBT (n = 42) or CAP (n = 27) for tetralogy of Fallot (n = 21), pulmonary atresia (n = 25), severe pulmonary valve stenosis (n = 22), and 2-stage repair of transposition of the great arteries (n = 1). The groups were similar with regard to age, weight, pulmonary artery diameter, and preoperative saturations. Postoperative mortality was 3 after CAP (11.1%) versus 1 after MBT (2.4%; P = .0203). Shunt size/weight index was comparable for both groups. MBTs had shorter surgical times (P = .002), required less inotropes (inotropic index, 103 ± 18 vs 889 ± 199; P = .0069), less blood product transfusions (P = .01), and had shorter duration of ventilation (P = .026) and intensive care unit (ICU) stay (P = .042). Children with MBTs had higher saturations at hospital discharge (P = .018). Prior to complete repair, 2 patients with a CAP and 10 patients after an MBT needed pulmonary artery dilation or stent implantation (P = .23). At the time of complete repair and shunt takedown, 3 MBT patients needed surgical patch augmentation of the pulmonary artery. The MBT is a safer and more expeditious operation and more frequently avoids cardiopulmonary bypass. Patients require less inotropes, blood products, and ICU time but may require more interventional therapy to treat pulmonary artery stenosis in the interval to complete repair. Surgical treatment of shunt-related pulmonary artery distortion may be addressed at the time of complete repair.

KIF5C: a new binding partner for protein kinase CK2 with a preference for the CK2alpha' subunit.

Protein kinase CK2 is a highly conserved serine/threonine kinase that is ubiquitously expressed in eukaryotic cells. CK2 is a constitutively active tetrameric enzyme composed of two catalytic alpha and/or alpha'-subunits and two regulatory beta-subunits. There is increasing evidence that the individual subunits may have independent functions and that they are asymmetrically distributed inside the cell. To gain a better understanding of the functions of the individual subunits, we employed a yeast-two-hybrid screen with CK2alpha and CK2alpha'. We identified the motor neuron protein KIF5C as a new binding partner for CK2. The interaction found in the yeast-two-hybrid screen was confirmed by co-sedimentation analysis on a sucrose density gradient and by co-immunoprecipitation analysis. Pull-down experiments and surface plasmon resonance spectrometry revealed a direct binding of KIF5C to CK2alpha'. Co-localization studies with neuroblastoma cells, bone marrow and with primary neurons confirmed the biochemical analysis that KIF5C preferentially bound to CK2alpha'.

Protein kinase CK2 interacts with the splicing factor hPrp3p.

Numerous signalling pathways in cells are influenced by the ubiquitous Ser/Thr protein kinase CK2. Protein kinase CK2 is composed of two regulatory beta-subunits and two catalytic alpha- or alpha'-subunits. Several of the known CK2 substrates are proteins known to regulate transcriptional events. Here, we describe that protein kinase CK2 interacts with the splicing factor hPrp3p, which is important for the assembly of the spliceosome. In a two-hybrid screen hPrp3p is exclusively bound to the catalytic alpha- or alpha'-subunits of CK2 but not to the regulatory beta-subunit. The interaction was confirmed by coimmunoprecipitation experiments in vitro and in vivo. Moreover, both proteins colocalized in nuclear speckles which is typical for splicing factor compartments within the nucleus. Phosphorylation experiments revealed that hPrp3p is also a substrate of protein kinase CK2. The main phosphorylation site was mapped to C-terminal residues. In vitro and in vivo splicing assays showed that the splicing activity is significantly influenced by the CK2-hPrp3p interaction. Thus, these data showed that CK2 is involved in the regulation of RNA processing.

[Initial experiences with adjuvant locoregional radioimmunotherapy using 131I-labeled monoclonal antibodies against tenascin (BC-4) for treatment of glioma (WHO III and IV)]. / Initiale Erfahrungen mit der adjuvanten lokoregionalen Radioimmuntherapie mit 131I-markierten monoklonalen Tenascin-Antikörpern (BC-4) bei Patienten mit Gliom (WHO III und IV).

AIM: None of the established treatments (surgery, radiotherapy, chemotherapy) for malignant glioma has improved its very poor prognosis. Adjuvant locoregional radio-immunotherapy (RIT) represents a new therapeutic approach. We present our initial experience with this therapeutic tool with respect to adverse effects, biokinetics and clinical follow-up. METHODS: Following surgery and radiotherapy, 12 patients with glioma (4, WHO stage III; 8, WHO stage IV) underwent 1-5 RIT-cycles (average dose 1100 MBq 131labelled monoclonal BC-4 antibodies) at six week intervals. Follow-up included serial FDG-PET and MRI investigations. Evaluation of biokinetics included whole body scans, together with analysis of blood, urine and fluid from the tumor cavity. RESULTS: Following RIT, four patients experienced temporary seizures, which, in one case, were associated with temporary aphasia. Eight patients developed HAMA (human anti-mouse antibodies) during follow-up. Mean biologic half-life of the radiopharmaceutical in the resection cavity was 3.9 d (range: 1.0-10.2 d) and remained stable intraindividually during further RIT-cycles. The antibody/radionuclide conjugate remained stable in the tumor cavity for at least 5 d. Median survival presently stands at 18.5 months compared to 9.7 months in a historical patient group (n = 89) undergoing conventional therapeutic strategies. Five patients show no signs of recurrence. In three patients with post-surgical evidence of residual tumor, one patient showed partial remission, one stable disease, and one progressive disease during RIT. Four patients without evidence of residual tumor mass at the beginning of RIT developed recurrence during therapy. CONCLUSIONS: Initial experience demonstrates that locoregional RIT is a well tolerated treatment modality that may represent a promising new approach in the management of patients with malignant glioma. Advantages of local application include passage of the blood-brain barrier, high concentration of activity within the resection cavity and low systemic toxicity.

Specific localization of the catalytic subunits of protein kinase CK2 at the centrosomes.

The protein kinase CK2 holoenzyme is composed of two regulatory beta subunits and two catalytic alpha or alpha' subunits. Although experimental evidence for involvement of the enzyme in the regulation of cell proliferation is accumulating, the exact mechanism of its action is still unclear. The subcellular localization of the enzyme may be a key to its function. We have recently shown that the CK2 holoenzyme is tightly associated with the Golgi complex and the endoplasmic reticulum. Centrosomes, which organize spindle formation during the cell cycle and microtubule cytoskeleton formation and, thereby, the location and orientation of different organelles in the cell, are in close vicinity to the Golgi complex. Because several kinases and phosphatases have been described to regulate the functions of the centrosome, we analysed the association of CK2 with these organelles. Using biochemical cell fractionation and coimmunoprecipitation, we never found the holoenzyme but only the catalytic alpha subunits associated with the centrosome. These data were confirmed by immunoelectron microscopy. Thus, the present data point to a particular role of the catalytic alpha and alpha' subunit of protein kinase CK2, which may be different from their roles in the holoenzyme.

[Normal pressure hydrocephalus--from clinical picture to diagnosis. Help by early shunt placement]. / Normaldruckhydrozephalus--Vom klinischen Bild zur Diagnose. Hilfe durch rechtzeitige Shuntanlage.

Normal pressure hydrocephalus (NPH), which may either be idiopathic or occur secondarily, is responsible for approximately 6% of the cases of dementional syndromes in the elderly. Although its diagnosis still remains a problem, every attempt should be made to identify patients with clinical symptoms arising from NPH, since the placement of a shunt can result in a much improved quality of life. Candidates for surgical treatment must be selected with care, since the transient peri-operative morbidity may be as high as 7%.

Downregulation of the cdc2/cyclin B protein kinase activity by binding of p53 to p34(cdc2).

We previously found that p53 binds to the catalytic subunit of the p34(cdc2)/cyclin B1-kinase. In the present study we analyzed the functional consequences of this interaction. Binding of wild-type p53 to p34(cdc2)/cyclin B1 results in a significant decrease of its histone H1 kinase activity. Binding of p53 to the kinase is a prerequisite for the inhibition because a mutant p53 which lacks the binding region fails to influence the enzymatic activity. Furthermore, by using C-terminal fragments of p53 it became obvious that also some other structural elements in the N-terminal region are necessary for the inhibitory effect. Our present study provides evidence that p53 might regulate cell-cycle checkpoints not only on the transcriptional level but also by binding to the cell-cycle regulating kinase p34(cdc2).

Wild-type p53 inhibits protein kinase CK2 activity.

The growth suppressor protein p53 and the protein kinase CK2 are both implicated in cellular growth regulation. We previously found that p53 binds to protein kinase CK2 via its regulatory beta-subunit. In the present study, we analyzed the consequences of the binding of p53 to CK2 for the enzymatic activity of CK2 in vitro and in vivo. We found that the carboxy-terminus of p53 which is a potent transforming agent stimulated CK2 activity whereas full length wild-type p53 which is a growth suppressor inhibited the activity of protein kinase CK2. Inhibition of protein kinase CK2 by p53 was dose-dependent and was seen for various CK2 substrates. Experiments with heat-denatured p53 and the conformational mutant p53(R175H) revealed that an intact conformation of p53 seemed to be necessary. Transfection of wild-type and of mutant p53 into p53-/- cells showed that the inhibition of p53 on CK2 activity was also detectable in intact cells and specific for wild-type p53 indicating that the growth suppressing function of p53 might at least be partially achieved by down-regulation of protein kinase CK2.

Binding of the growth suppressor p53 protein to the cell cycle regulator phosphatase cdc25C.

Human p53 is a growth suppressor which not only functions in mammalian cells but also in fission yeast. It was previously shown that the cell cycle regulating phosphatase cdc25C suppresses the p53 induced growth arrest in fission yeast. In the present study we analysed the mechanism of this suppression. We found that cdc25C directly interacts with p53. By using different deletion mutants the binding region was narrowed down on the polypeptide chain of p53 to amino acids 287-340. To test the functional significance we analysed the effect of this interaction on the DNA binding activity of p53. As shown by band shift experiments binding of cdc25C to p53 does not modify the DNA binding activity of p53. Our data suggest that the observed suppression of the p53 induced growth arrest by cdc25C might be achieved by direct binding of cdc25C to the C-terminus of p53.

Motion determination in actin filament fluorescence images with a spatio-temporal orientation analysis method.

We present a novel approach of automatically measuring motion in series of microscopic fluorescence images. As a differential method, the three-dimensional structure tensor technique is used to calculate the displacement vector field for every image of the sequence, from which the velocities are subsequently derived. We have used this method for the analysis of the movement of single actin filaments in the in vitro motility assay, where fluorescently labeled actin filaments move over a myosin decorated surface. With its fast implementation and subpixel accuracy, this approach is, in general, very valuable for analyzing dynamic processes by image sequence analysis.

Growth hormone treatment of breeding bulls used for artificial insemination improves fertilization rates.

To evaluate new therapeutical concepts for male subfertility, we tested the effects of exogenous recombinant bovine growth hormone (rbGH) on various endocrine and metabolic parameters both in blood and in seminal plasma of bulls. Sperm quality was assessed morphometrically and by monitoring the number of successful artificial inseminations (AIs) defined as non-return rates (NRR). Aliquots of 450 semen samples were used from each bull and each experimental period (4 wk before, 14 weeks during and 6 wk after treatment). Six out of ten sires (average age 8.4 years) were treated every two weeks with 640-mg depot formulated rbGH (Eli Lilly). Four bulls received vehicle only. Blood plasma bGH, IGF-I, insulin and glucose concentrations were increased with rbGH treatment. In seminal plasma there was no effect of rbGH treatment on fructose and citrate or on testosterone concentrations. With one exception, rbGH-treated bulls had greater IGFBP-3 concentrations in seminal plasma. Motility of spermatozoa after freezing and thawing was increased compared with pretreatment rates. Most interestingly, the number of successful AIs was increased by an average of 6.0% NRR when ejaculates from rbGH-treated bulls were used.

Binding domain for p21(WAF1) on the polypeptide chain of the protein kinase CK2 beta-subunit.

Protein kinase CK2 is a ubiquitous serine/threonine kinase which is involved in many proliferation-related processes in the cell. It is composed of two regulatory beta-subunits and two catalytic alpha-subunits. Its regulation still remains mysterious in spite of many years of intense research. One of its regulators is the cdk inhibitory molecule p21(WAF1)-a protein which is expressed in situations of genotoxic stress. p21(WAF1) binds to the beta-subunit of CK2 and inhibits the activity of CK2. Using deletion mutants of CK2 beta as well as a peptide library consisting of 15-amino-acid-long peptides derived from the polypeptide chain of CK2 beta we mapped the binding region for p21(WAF1) on the polypeptide chain of CK2 beta. We localized an amino-terminal and a carboxy-terminal binding domain. Binding of p21(WAF1) to both regions of the CK2 beta-subunit interferes with the phosphotransferase activity of the CK2 holoenzyme.