julilili42/papers huggingface dataset:
Up-Regulation of Excitatory Amino Acid Transporters EAAT3 and EAAT4 by Lithium Sensitive Glycogen Synthase Kinase GSK3ß
2016-01-01

Background: Cellular uptake of glutamate by the excitatory amino-acid transporters (EAATs) decreases excitation and thus participates in the regulation of neuroexcitability. Kinases impacting on neuronal function include Lithium-sensitive glycogen synthase kinase GSK3ß. The present study thus explored whether the activities of EAAT3 and/or EAAT4 isoforms are sensitive to GSK3ß. Methods: cRNA encoding wild type EAAT3 (SLC1A1) or EAAT4 (SLC1A6) was injected into Xenopus oocytes without or with additional injection of cRNA encoding wild type GSK3ß or the inactive mutant K85AGSK3ß. Dual electrode voltage clamp was performed in order to determine glutamate-induced current (IEAAT). Results: Appreciable IEAAT was observed in EAAT3 or EAAT4 expressing but not in water injected oocytes. IEAAT was significantly increased by coexpression of GSK3ß but not by coexpression of K85AGSK3ß. Coexpression of GSK3ß increased significantly the maximal IEAAT in EAAT3 or EAAT4 expressing oocytes, without significantly modifying apparent affinity of the carriers. Lithium (1 mM) exposure for 24 hours decreased IEAAT in EAAT3 and GSK3ß expressing oocytes to values similar to IEAAT in oocytes expressing EAAT3 alone. Lithium did not significantly modify IEAAT in oocytes expressing EAAT3 without GSK3ß. Conclusions: Lithium-sensitive GSK3ß is a powerful regulator of excitatory amino acid transporters EAAT3 and EAAT4.
Generation of an isogenic, gene-corrected control cell line of the spinocerebellar ataxia type 2 patient-derived iPSC line H196
2016-01-01

Spinocerebellar ataxia type 2 (SCA2) is a neurodegenerative disease primarily affecting the cerebellum. Very little is known about the molecular mechanisms underlying the disease and, to date, no cure or treatment is available. We have successfully generated bona fide induced pluripotent stem cell (iPSC) lines of SCA2 patients in order to study a disease-specific phenotype. Here, we demonstrate the gene correction of the iPSC line H196 clone 7 where we have exchanged the expanded CAG repeat of the ATXN2 gene with the normal length found in healthy alleles. This gene corrected cell line will provide the ideal control to model SCA2 by iPSC technology.
A Systematic Perspective on Intergenerational Learning: Theoretical and Empirical Findings
2016-01-01

Intergenerational learning is a growing field of practice across Europe. Against a background of demographic and social changes within European countries and the subsequently altering learning processes between different generations in families, communities, and workplaces, educational organizations face the challenge of providing learning opportunities for the different generations. This paper presents research related to the development of intergenerational learning programs in German adult education organizations. Within that project, adult educators from different organizations took part in a training module on intergenerational learning and subsequently created intergenerational learning programs in their own organizations. In this paper we provide insight into the training program of the participating adult educators, as well as into the results of our empirical study. First, we present the systematic framework of intergenerational learning that was used in the training module to initiate the development of practical intergenerational programs. We then present our research design and the results of the qualitative study. Finally, we discuss these results in terms of theory.
Personalized Epigenetics
2016-01-01

Personalized epigenetics identifies epigenetic alterations in individual patients as a predisposition for diseases or, if a disease is already present, their influence on disease progression and therapeutic outcome. This review gives a brief overview of different epigenetic layers that will impact personalized medicine, focused on DNA and histone modifications. Examples of how epigenetic drivers affect the course of diseases will be discussed with an emphasis on cancer; however, many of these concepts are applicable to other diseases. In contrast to genetic mutations, alterations in the epigenome are much more dynamic. Environmental influences or even aging constantly modify existing epigenetic traits. Furthermore, therapies produce a considerable selection pressure in epigenetic marks that can contribute to therapy resistance. The increasing knowledge of the dynamic epigenetic landscape along with the possibility to reverse epigenetic modifications makes this an attractive field for drug development. However, several examples show that the outcome of epigenetic interventions is context dependent. Therefore, new therapeutic principles have to carefully consider the interactions between epigenetic writers, readers, erasers, and chromatin modifying complexes. To keep pace with the huge technological advances and the rapidly increasing knowledge in biomedicine, new collaborations have to be implemented to include personalized epigenetics in clinical decision-making.
Partial Reversal of Tissue Calcification and Extension of Life Span following Ammonium Nitrate Treatment of Klotho-Deficient Mice
2016-01-01

&lt;b&gt;&lt;i&gt;Background/Aims: &lt;/i&gt;&lt;/b&gt;Klotho is required for the inhibitory effect of FGF23 on 1,25(OH)&lt;sub&gt;2&lt;/sub&gt;D&lt;sub&gt;3&lt;/sub&gt; formation and Klotho-hypomorphic mice (&lt;i&gt;kl/kl&lt;/i&gt;) suffer from severe tissue calcification due to excessive 1,25(OH)&lt;sub&gt;2&lt;/sub&gt;D&lt;sub&gt;3&lt;/sub&gt; formation with subsequent increase of Ca&lt;sup&gt;2+&lt;/sup&gt; and phosphate concentrations and stimulation of osteogenic signaling. The excessive tissue calcification dramatically accelerates aging and leads to premature death of the animals. Osteogenic signaling in those mice is disrupted by treatment with NH&lt;sub&gt;4&lt;/sub&gt;Cl, which prevents tissue calcification and early death of &lt;i&gt;kl/kl&lt;/i&gt; mice. The present study explored whether the beneficial effects of NH&lt;sub&gt;4&lt;/sub&gt;Cl treatment could be mimicked by NH&lt;sub&gt;4&lt;/sub&gt;NO&lt;sub&gt;3&lt;/sub&gt; treatment. &lt;b&gt;&lt;i&gt;Methods: &lt;/i&gt;&lt;/b&gt;The &lt;i&gt;kl/kl&lt;/i&gt; mice had free access to tap water either without or with addition of NH&lt;sub&gt;4&lt;/sub&gt;NO&lt;sub&gt;3&lt;/sub&gt; (0.28 M) starting with the mating of the parental generation. Calcification of trachea, lung, kidney, stomach, heart and vessels was visualized by histology with von Kossa staining. Plasma phosphate concentration was determined utilizing photometry, blood gas and electrolytes utilizing a blood Gas and Chemistry Analysis System and plasma 1,25(OH)&lt;sub&gt;2&lt;/sub&gt;D&lt;sub&gt;3&lt;/sub&gt; concentration with ELISA. &lt;b&gt;&lt;i&gt;Results: &lt;/i&gt;&lt;/b&gt;In untreated &lt;i&gt;kl/kl&lt;/i&gt; mice plasma 1,25(OH)&lt;sub&gt;2&lt;/sub&gt;D&lt;sub&gt;3&lt;/sub&gt; and phosphate concentrations were elevated, and the mice suffered from marked calcification of all tissues analyzed. Untreated &lt;i&gt;kl/kl&lt;/i&gt; mice further suffered from respiratory acidosis due to marked lung emphysema. NH&lt;sub&gt;4&lt;/sub&gt;NO&lt;sub&gt;3&lt;/sub&gt;-treatment decreased both, blood pCO&lt;sub&gt;2&lt;/sub&gt; and HCO&lt;sub&gt;3&lt;/sub&gt;&lt;sup&gt;-&lt;/sup&gt;, decreased calcification of trachea, lung, kidney, stomach, heart and vessels and increased the life span of &lt;i&gt;kl/kl&lt;/i&gt; mice more than 1.7-fold (&amp;#x2642;) or 1.6-fold (&amp;#x2640;) without significantly affecting extracellular pH or plasma concentrations of 1,25(OH)&lt;sub&gt;2&lt;/sub&gt;D&lt;sub&gt;3&lt;/sub&gt;, Ca&lt;sup&gt;2+&lt;/sup&gt;, phosphate, Na&lt;sup&gt;+&lt;/sup&gt;, and K&lt;sup&gt;+&lt;/sup&gt;. &lt;b&gt;&lt;i&gt;Conclusions: &lt;/i&gt;&lt;/b&gt;NH&lt;sub&gt;4&lt;/sub&gt;NO&lt;sub&gt;3&lt;/sub&gt;-treatment turns respiratory acidosis into metabolic acidosis and mitigates calcification thus leading to a substantial extension of &lt;i&gt;kl/kl&lt;/i&gt; mice survival.
Decrease of Store-Operated Ca2+ Entry and Increase of Na+/Ca2+ Exchange by Pharmacological JAK2 Inhibition
2016-01-01

Background/Aims: Cell proliferation and migration are regulated by cytosolic Ca2+ activity ([Ca2+]i). Mechanisms modifying [Ca2+]i include store-operated Ca2+-entry (SOCE) accomplished by the pore-forming ion channel unit Orai1 and its regulator STIM1, as well as Ca2+ extrusion by Na+/Ca2+ exchanger NCX1. Kinases participating in the orchestration of cell proliferation include the Janus activated kinase JAK2. The present study explored the impact of pharmacological JAK2 inhibition on SOCE and Na+/Ca2+ exchange. Methods: MCF-7 breast carcinoma cells were studied in the absence and presence of the JAK2 inhibitors TG101348 (250 nM - 1 μM) or of AG490 (20 - 40 μM). Transcript levels were quantified with RT-PCR, protein abundance with immunoblotting, [Ca2+]i with Fura-2-fluorescence, SOCE from increase of [Ca2+]i following Ca2+ re-addition after Ca2+-store depletion with sarcoendoplasmatic Ca2+-ATPase (SERCA) inhibitor thapsigargin (1 μM), and Na+/Ca2+ exchanger activity from increase of [Ca2+]i as well as Ca2+ current in whole cell patch clamp following extracellular Na+ removal. Migratory activity was determined by a wound healing assay. Results: JAK2 inhibitor TG101348 (1 μM) decreased Orai1 and STIM1 protein abundance, increased NCX1 transcript levels, decreased Ca2+ release from intracellular stores, decreased SOCE, increased Ca2+ entry as well as Ca2+-current following extracellular Na+-removal, and decreased migration. Similar effects on Ca2+ release, SOCE, and Ca2+-entry following extracellular Na+-removal were observed following treatment with AG490. Conclusion: The present observations disclose a novel powerful mechanism regulating intracellular Ca2+ release, cellular Ca2+ entry, cellular Ca2+ extrusion and cell migration.
Stimulation of Suicidal Erythrocyte Death by Ceritinib-Treatment of Human Erythrocytes
2016-01-01

Background/Aims: The anaplastic lymphoma kinase (ALK) inhibitor ceritinib is utilized for the treatment of ALK positive non-small cell lung carcinoma. Side effects of the drug include decrease of blood hemoglobin concentration. Possible causes of anemia include stimulation of suicidal erythrocyte death or eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Signaling of eryptosis includes increase of cytosolic Ca2+ activity ([Ca2+]i), oxidative stress, ceramide, staurosporine sensitive protein kinase C, SB203580 sensitive p38 kinase, D4476 sensitive casein kinase 1, and zVAD sensitive caspases. The present study explored, whether ceritinib induces eryptosis and, if so, to shed light on the cellular mechanisms involved. Methods: Phosphatidylserine exposure at the cell surface was estimated from annexin-V-binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, ROS formation from DCFDA dependent fluorescence, and ceramide abundance utilizing specific antibodies. Results: A 48 hours exposure of human erythrocytes to ceritinib (1 μg/ml) significantly increased the percentage of annexin-V-binding cells, significantly decreased forward scatter, significantly increased Fluo3-fluorescence, but did not significantly modify DCFDA fluorescence or ceramide abundance. The effect of ceritinib on annexin-V-binding was significantly blunted but not abolished by removal of extracellular Ca2+, by the kinase inhibitors staurosporine (1 μM), SB203580 (2 μM) and D4476 (10 μM), as well as by caspase inhibitor zVAD (10 μM). Conclusions: Ceritinib triggers cell shrinkage and phospholipid scrambling of the erythrocyte cell membrane, an effect at least in part due to Ca2+ entry, as well as activation of kinases and Caspases.
Inter-Rater Reliability of Neck Reflex Points in Women with Chronic Neck Pain
2016-01-01

&lt;b&gt;&lt;i&gt;Background:&lt;/i&gt;&lt;/b&gt; Neck reflex points (NRP) are tender soft tissue areas of the cervical region that display reflectory changes in response to chronic inflammations of correlated regions in the visceral cranium. Six bilateral areas, NRP C0, C1, C2, C3, C4 and C7, are detectable by palpating the lateral neck. We investigated the inter-rater reliability of NRP to assess their potential clinical relevance. &lt;b&gt;&lt;i&gt;Methods:&lt;/i&gt;&lt;/b&gt; 32 consecutive patients with chronic neck pain were examined for NRP tenderness by an experienced physician and an inexperienced medical student in a blinded design. A detailed description of the palpation technique is included in this section. Absence of pain was defined as pain index (PI) = 0, slight tenderness = 1, and marked pain = 2. Findings were evaluated either by pair-wise Cohen's kappa (&amp;#x0138;) or by percentage of agreement (PA). &lt;b&gt;&lt;i&gt;Results:&lt;/i&gt;&lt;/b&gt; Examiners identified 40% and 41% of positive NRP, respectively (PI &gt; 0, physician: 155, student: 157) with a slight preference for the left side (1.2:1). The number of patients identified with &gt;6 positive NRP by the examiners was similar (13 vs. 12 patients). &amp;#x0138; values ranged from 0.52 to 0.95. The overall kappa was &amp;#x0138; = 0.80 for the left and &amp;#x0138; = 0.74 for the right side. PA varied from 78.1% to 96.9% with strongest agreement at NRP C0, NRP C2, and NRP C7. Inter-rater agreement was independent of patients' age, gender, body mass index and examiner's experience. &lt;b&gt;&lt;i&gt;Conclusion:&lt;/i&gt;&lt;/b&gt; The high reproducibility suggests the clinical relevance of NRP in women.
A bounded p-norm approximation of max-convolution for sub-quadratic Bayesian Inference on additive factors
2016-01-01

Max-convolution is an important problem closely resembling standard convolution; as such, max-convolution occurs frequently across many fields. Here we extend the method with fastest known worst-case runtime, which can be applied to nonnegative vectors by numerically approximating the Chebyshev norm || ċ ||∞, and use this approach to derive two numerically stable methods based on the idea of computing p-norms via fast convolution: The first method proposed, with runtime in O(k log(k) log(log(k))) (which is less than 18k log(k) for any vectors that can be practically realized), uses the p-norm as a direct approximation of the Chebyshev norm. The second approach proposed, with runtime in O(k log(k)) (although in practice both perform similarly), uses a novel null space projection method, which extracts information from a sequence of p-norms to estimate the maximum value in the vector (this is equivalent to querying a small number of moments from a distribution of bounded support in order to estimate the maximum). The p-norm approaches are compared to one another and are shown to compute an approximation of the Viterbi path in a hidden Markov model where the transition matrix is a Toeplitz matrix; the runtime of approximating the Viterbi path is thus reduced from O(nk2) steps to O(nk log(k)) steps in practice, and is demonstrated by inferring the U.S. unemployment rate from the S&P 500 stock index.
On Retinal Gene Therapy
2016-01-01

Mutations in a large number of genes cause retinal degeneration and blindness with no cure currently available. Retinal gene therapy has evolved over the last decades to become a promising new treatment paradigm for these rare disorders. This article reflects on the ideas and concepts arising from basic science towards the translation of retinal gene therapy into the clinical realm. It describes the advances and present thinking on the efficacy of current clinical trials and discusses potential roadblocks and solutions for the future of retinal gene therapy.
β-Klotho as a Negative Regulator of the Peptide Transporters PEPT1 and PEPT2
2016-01-01

Background/Aims: β-Klotho, a transmembrane protein expressed in several tissues including the brain and the kidney, is critically important for inhibition of 1,25(OH)2D3 formation by FGF23. The extracellular domain of Klotho protein could be cleaved off, thus being released into blood or cerebrospinal fluid. Soluble klotho is a β-glucuronidase participating in the regulation of several ion channels and carriers. The present study explored the effect of β-Klotho protein on the peptide transporters PEPT1 and PEPT2. Methods: cRNA encoding PEPT1 or PEPT2 was injected into Xenopus laevis oocytes and glycine-glycine (2 mM)-induced inward current (IGly) taken as measure of glycine-glycine transport. Measurements were made without or with prior 24 h treatment with soluble β-Klotho protein (30 ng/ml) in the absence and presence of β-glucuronidase inhibitor D-saccharic acid 1,4-lactone monohydrate (DSAL,10 μM). Ussing chamber experiments were employed to determine electrogenic peptide transport across intestinal epithelia of klotho deficient (kl-/-) and corresponding wild type (kl+/+) mice. Results: IGly was observed in PEPT1 and in PEPT2 expressing oocytes but not in water injected oocytes. In both, PEPT1 and PEPT2 expressing oocytes IGly was significantly decreased by treatment with soluble β-Klotho protein. As shown for PEPT1, β-klotho protein decreased significantly the maximal transport rate without significantly modifying the affinity of the carrier. The effect of β-Klotho on PEPT1 was reversed by DSAL. Intestinal IGly was significantly larger in kl-/- than in kl+/+ mice. Conclusion: β-Klotho participates in the regulation of the peptide transporters PEPT1 and PEPT2.
Novel KCNQ3 Mutation in a Large Family with Benign Familial Neonatal Epilepsy: A Rare Cause of Neonatal Seizures
2016-01-01

Benign familial neonatal seizures (BFNS) present a rare familial epilepsy syndrome caused by genetic alterations in the voltage-gated potassium channels Kv7.2 and Kv7.3, encoded by &lt;i&gt;KCNQ2&lt;/i&gt; and &lt;i&gt;KCNQ3.&lt;/i&gt; While most BFNS families carry alterations in &lt;i&gt;KCNQ2&lt;/i&gt;, mutations in &lt;i&gt;KCNQ3&lt;/i&gt; appear to be less common. Here, we describe a family with 6 individuals presenting with neonatal focal and generalized seizures. Genetic testing revealed a novel &lt;i&gt;KCNQ3&lt;/i&gt; variant, c.835G&gt;T, cosegregating with seizures in 4 tested individuals. This variant results in a substitution of the highly conserved amino acid valine localized within the pore-forming transmembrane segment S5 (p.V279F). Functional investigations in &lt;i&gt;Xenopus laevis&lt;/i&gt; oocytes revealed a loss of function, which supports p.V279F as a pathogenic mutation. When p.V279F was coexpressed with the wild-type (WT) Kv7.2 subunits, the resulting potassium currents were about 10-fold reduced compared to the WT Kv7.3 and Kv7.2 coexpression. Genotype-phenotype correlation shows an incomplete penetrance of p.V279F. Response to antiepileptic treatment was variable, but evaluation of treatment response remained challenging due to the self-limiting character of the disease. The identification of the pathogenic variant helped to avoid unnecessary investigations in affected family members and allowed guided therapy.
Generation of an isogenic, gene-corrected control cell line of the spinocerebellar ataxia type 2 patient-derived iPSC line H266
2016-01-01

Spinocerebellar ataxia type 2 (SCA2) is a neurodegenerative disease primarily affecting the cerebellum. Very little is known about the molecular mechanisms underlying the disease and, to date, no cure or treatment is available. We have successfully generated bona fide induced pluripotent stem cell (iPSC) lines of SCA2 patients in order to study a disease-specific phenotype. Here, we demonstrate the gene correction of the iPSC line H266 clone 10 where we have exchanged the expanded CAG repeat of the ATXN2 gene with the normal length found in healthy alleles. This gene corrected cell line will provide the ideal control to model SCA2 by iPSC technology.
1α,25(OH)2D3 Induces Actin Depolymerization in Endometrial Carcinoma Cells by Targeting RAC1 and PAK1
2016-01-01

Background: Cell proliferation and motility require actin reorganization, which is under control of various signalling pathways including ras-related C3 botulinum toxin substrate 1 (RAC1), p21 protein-activated kinase 1 (PAK1) and actin related protein 2 (ARP2). Tumour cell proliferation is modified by 1α,25-Dihydroxy-Vitamin D3 (1α,25(OH)2D3), a steroid hormone predominantly known for its role in calcium and phosphorus metabolism. The present study explored whether 1α,25(OH)2D3 modifies actin cytoskeleton in Ishikawa cells, a well differentiated endometrial carcinoma cell line. Methods: To this end, actin cytoskeleton was visualized by confocal microscopy. Globular over filamentous actin ratio was determined utilizing Western blotting and flow cytometry, transcript levels by qRT-PCR and protein abundance by immunoblotting. Results: A 24 hour treatment with 1α,25(OH)2D3 (100 nM) significantly decreased RAC1 and PAK1 transcript levels and activity, decreased ARP2 protein levels and depolymerized actin. The effect of 1α,25(OH)2D3 on actin polymerization was mimicked by pharmacological inhibition of RAC1 and PAK1. Conclusions: 1α,25(OH)2D3 leads to disruption of RAC1 and PAK1 activity with subsequent actin depolymerization of endometrial carcinoma cells.
Regulation of the growth, morphology, mechanical properties and biocompatibility of natural polysaccharide-based multilayers by Hofmeister anions
2016-01-01

Herein the optimization of the physicochemical properties and surface biocompatibility of polyelectrolyte multilayers of the natural, biocompatible and biodegradable, linear polysaccharides hyaluronan and chitosan by Hofmeister anions was systematically investigated.
How Confocal Is Confocal Raman Microspectroscopy on the Skin? Impact of Microscope Configuration and Sample Preparation on Penetration Depth Profiles
2016-01-01

&lt;b&gt;&lt;i&gt;Background/Aims:&lt;/i&gt;&lt;/b&gt; The aim of the study was to elucidate the effect of sample preparation and microscope configuration on the results of confocal Raman microspectroscopic evaluation of the penetration of a pharmaceutical active into the skin (depth profiling). &lt;b&gt;&lt;i&gt;Methods:&lt;/i&gt;&lt;/b&gt; Pig ear skin and a hydrophilic formulation containing procaine HCl were used as a model system. The formulation was either left on the skin during the measurement, or was wiped off or washed off prior to the analysis. The microscope configuration was varied with respect to objectives and pinholes used. &lt;b&gt;&lt;i&gt;Results:&lt;/i&gt;&lt;/b&gt; Sample preparation and microscope configuration had a tremendous effect on the results of depth profiling. Regarding sample preparation, the best results could be observed when the formulation was washed off the skin prior to the analysis. Concerning microscope configuration, the use of a 40 × 0.6 numerical aperture (NA) objective in combination with a 25-μm pinhole or a 100 × 1.25 NA objective in combination with a 50-μm pinhole was found to be advantageous. &lt;b&gt;&lt;i&gt;Conclusion:&lt;/i&gt;&lt;/b&gt; Complete removal of the sample from the skin before the analysis was found to be crucial. A thorough analysis of the suitability of the chosen microscope configuration should be performed before acquiring concentration depth profiles.
Outcome, Treatment, and Treatment Failures in Patients Suffering Localized Embryonal Paratesticular Rhabdomyosarcoma
2016-01-01

The outcome of PTRMS patients is excellent and is hardly improvable. Positive predictors for outcome are age younger than 10 years and tumor size less than 5 cm. Primary LN sampling seems to have no impact on the EFS and should not be recommended. Inadequate surgery can be avoided by treatment in specialized centers. Secondary retroperitoneal LN dissection revealed a high number of unnecessary procedures, but patients with positive LN had a poor prognosis and require additional local therapy.