Refining Genotypes and Phenotypes in KCNA2-Related Neurological Disorders

Pathogenic variants in KCNA2, encoding for the voltage-gated potassium channel K_v1.2, have been identified as the cause for an evolving spectrum of neurological disorders. Affected individuals show early-onset developmental and epileptic encephalopathy, intellectual disability, and movement disorders resulting from cerebellar dysfunction. In addition, individuals with a milder course of epilepsy, complicated hereditary spastic paraplegia, and episodic ataxia have been reported. By analyzing phenotypic, functional, and genetic data from published reports and novel cases, we refine and further delineate phenotypic as well as functional subgroups of KCNA2-associated disorders. Carriers of variants, leading to complex and mixed channel dysfunction that are associated with a gain- and loss-of-potassium conductance, more often show early developmental abnormalities and an earlier onset of epilepsy compared to individuals with variants resulting in loss- or gain-of-function. We describe seven additional individuals harboring three known and the novel KCNA2 variants p.(Pro407Ala) and p.(Tyr417Cys). The location of variants reported here highlights the importance of the proline(405)–valine(406)–proline(407) (PVP) motif in transmembrane domain S6 as a mutational hotspot. A novel case of self-limited infantile seizures suggests a continuous clinical spectrum of KCNA2-related disorders. Our study provides further insights into the clinical spectrum, genotype–phenotype correlation, variability, and predicted functional impact of KCNA2 variants.     

Geometric adaption of biodegradable magnesium alloy scaffolds to stabilise biological myocardial grafts. Part I

Synthetic patch materials currently in use have major limitations, such as high susceptibility to infections and lack of contractility. Biological grafts are a novel approach to overcome these limitations, but do not always offer sufficient mechanical durability in early stages after implantation. Therefore, a stabilising structure based on resorbable magnesium alloys could support the biological graft until its physiologic remodelling. To prevent early breakage in vivo due to stress of non-determined forming, these scaffolds should be preformed according to the geometry of the targeted myocardial region. Thus, the left ventricular geometry of 28 patients was assessed via standard cardiac magnetic resonance imaging (MRI). The resulting data served as a basis for a finite element simulation (FEM). Calculated stresses and strains of flat and preformed scaffolds were evaluated. Afterwards, the structures were manufactured by abrasive waterjet cutting and preformed according to the MRI data. Finally, the mechanical durability of the preformed and flat structures was compared in an in vitro test rig. The FEM predicted higher durability of the preformed scaffolds, which was proven in the in vitro test. In conclusion, preformed scaffolds provide extended durability and will facilitate more widespread use of regenerative biological grafts for surgical left ventricular reconstruction.     

Game theoretic approaches to cost allocation in the dynamic total tardiness problem

We examine a dynamic Total Tardiness Problem (TTP) arising out of a decentralized job acquisition by several agents. Since the underlying static TTP is NP-hard, we first present a heuristic in order to solve the dynamic TTP without regard to the participation constraints of the agents. In the main part of the paper a cost allocation based on the results of the heuristic is developed by applying game theoretic concepts. Introducing the concept of a fair cost allocation according to Nash [1] in a special case, the results are subsequently transformed to suit the general situation. Two cost allocation schemes are discussed.     

The assessment of the umbilical blood flow of the surviving twin after the intrauterine death of the other twin

This paper summarizes our experience with Doppler velocimetry in survivors of intrauterine co-twin demise. In the first trimester, ten dichorionic deaths occurred; none of the survivors developed flow disorders. During the second trimester, there were three intrauterine demises, two of them were monochorionic and the survivors developed flow disorders: one presented transitory venous flow aberration, the other one an impaired development of diastolic flow. In the third trimester, two intrauterine deaths occurred. One case of twin to twin transfusion syndrome (TTTS) was complicated by the donor's death and the recipient showed a loss of diastolic flow. The second one happened during a dichorionic twin pregnancy. The survivor presented high systolic/diastolic daily ratio (S/D = 7.8).     

A new territory and its pioneer: opening up a dominant research stream for a translational research area

Scientometrics - An academic pioneer makes many trials to find relations and results that were unknown before. The verified relations and repeatable results the pioneer finds guide followers to...     

Multispectral fluorescence lifetime imaging by TCSPC

We present a fluorescence lifetime imaging technique with simultaneous spectral and temporal resolution. The technique is fully compatible with the commonly used multiphoton microscopes and nondescanned (direct) detection. An image of the back-aperture of the microscope lens is projected on the input of a fiber bundle. The input of the fiber bundle is circular, and the output is flattened to match the input slit of a spectrograph. The spectrum at the output of the spectrograph is projected on a 16-anode PMT module. For each detected photon, the encoding logics of the PMT module deliver a timing pulse and the number of the PMT channel in which the photon was detected. The photons are accumulated by a multidimensional time-correlated single photon counting (TCSPC) process. The recording process builds up a four-dimensional photon distribution over the times of the photons in the excitation pulse period, the wavelengths of the photons, and the coordinates of the scan area. The method delivers a near-ideal counting efficiency and is capable of resolving double-exponential decay functions. We demonstrate the performance of the technique for autofluorescence imaging of tissue.     

Optimization of a subclass of conjunctive queries

The optimization problem for a subclass of conjunctive queries which is formed by the union of the class of fan-out free queries and a subclass of typed fan-out queries is investigated. The typed fan-out queries in this class are obtained from simple tableaux by allowing atmost one attribute to violate the simple-tablau property. The optimization problem for several restricted subsets of typed fan-out queries is already known to be NP-hard. It is shown that the queries under consideration possess several useful properties which are then used to obtain an O(n ²) optimization algorithm based on the implication graph technique. The optimization of typed fan-out queries, obtained from simple tableaux by allowing atmost two attributes to violate the simple tableau property, is shown to be NP-hard. The optimization of simple tableaux in the presence of functional dependencies is also investigated and is shown to be NP-hard.Supported by DFG grant Bi 311/1-2; present address: School of Computer & Systems Sciences, Jawaharlal Nehru University, New Delhi 110067, India.Supported by NSF grant IRI-87-22886, AFOSR grant 88-0266 and a grant from IBM Corp.     

Fluorescence lifetime imaging by time-correlated single-photon counting

We present a time-correlated single photon counting (TCPSC) technique that allows time-resolved multi-wavelength imaging in conjunction with a laser scanning microscope and a pulsed excitation source. The technique is based on a four-dimensional histogramming process that records the photon density over the time of the fluorescence decay, the x-y coordinates of the scanning area, and the wavelength. The histogramming process avoids any time gating or wavelength scanning and, therefore, yields a near-perfect counting efficiency. The time resolution is limited only by the transit time spread of the detector. The technique can be used with almost any confocal or two-photon laser scanning microscope and works at any scanning rate. We demonstrate the application to samples stained with several dyes and to CFP-YFP FRET.     

Multi-dimensional fluorescence lifetime and FRET measurements

When and where proteins associate with each other in living cells are key questions in many biological research projects. One way to address these questions is to measure the extent of F?rster resonance energy transfer (FRET) between proteins that have been labeled with appropriate donor and acceptor fluorophores. When both proteins interact, donor and acceptor fluorophores are brought into close vicinity so that the donor can transmit a part of its excitation energy to the acceptor. As a result, both the intensity and the lifetime of the donor fluorescence decrease, whereas the intensity of the acceptor emission increases. This offers different approaches to determine FRET efficiency: One is to detect changes in the intensity of donor and acceptor emission, the other is to measure changes in the lifetime of the donor molecule. One important advantage of the fluorescence lifetime approach is that it allows to distinguish between free and associated donor molecules. However, like intensity measurements it lacks an intrinsic control ensuring that changes in the measured parameters are only due to FRET and not other quenching processes. Here, we show how this limitation can be overcome by spectrally resolved fluorescence lifetime measurements in the time domain. One technique is based on a streak camera system, the other technique is based on a time-correlated-single-photon-counting approach. Both approaches allow biologists to record both donor and acceptor fluorescence emitted by the sample in a single measurement.     

Decomposition of object-oriented database schemas

We specify a data model with object-oriented and logic-oriented features. The model is based on F-logic. For this model, we study a decomposition of a class, the counterpart to the well-known decomposition of a relation scheme under functional dependencies. For this decomposition of a class, the transformation pivoting is used. Pivoting transplants some attributes of the class to a newly generated class. This new class is a subclass of the result class of the so-called pivot attribute. The pivot attribute maintains the link between the original class and the new subclass. We identify the conditions under which input and output of pivoting are equivalent. Additionally, we show under which conditions a schema with functional dependencies can be recursively transformed into an equivalent one without non-key functional dependencies.