The Reckoning: The Return of Genomic Results to 1444 Participants Across the eMERGE3 Network

The goal of Electronic Medical Records and Genomics (eMERGE) Phase III Network was to return actionable sequence variants to 25,084 consenting participants from 10 different health care institutions across the United States. The purpose of this study was to evaluate system-based issues relating to the return of results (RoR) disclosure process for clinical grade research genomic tests to eMERGE3 participants

Untersuchung der Auswirkungen eines Polymorphismus im PDYN-Gen (rs1997794) auf die Furchtakquisition und -extinktion in einer fMRT-Studie unter Einbezug von Hautleitfähigkeitsveränderungen

Zusammenfassung Hintergrund: Untersuchungen der Vorgängerstudie von Bilkei-Gorzo et al. (2012) ergeben Hinweise, dass Prodynorphin (PDYN) einen wesentlichen Einfluss auf Akquisition und Extinktion von schmerzinduziertem Verhalten ausübt. Beim Menschen zeigt sich ein relevanter Einzelnukleotid-Polymorphismus bei rs1997794 (Promotorregion des PDYN-Gens), wobei homozygote T-Allelträger bei Schmerzreizen eine verstärkte Akquisitions- und eine verringerte Extinktionsleistung der Amygdala im Vergleich zu C-Allelträgern erkennen lassen. Ziel der vorliegenden Arbeit ist es, die humanexperimentellen Anteile der Vorgängerstudie mit größerer Stichprobe zu replizieren und PDYN-Effekte in der funktionellen Magnetresonanztomographie (fMRT) und Hautleitfähigkeitsmessung (engl. Skin Conductance Response, SCR) zu untersuchen. Methodik: Untersucht wird eine Stichprobe von 104 gesunden Erwachsenen (60 Frauen; Durchschnittsalter: 28,17 Jahre) mittels des von Bilkei-Gorzo et al. (2012) verwendeten fMRT-Paradigmas: Der dreiteilige Versuchsaufbau besteht aus Akquisition (ACQ), früher (EX1) und später Extinktion (EX2). Als unkonditionierter Reiz (UCS) dient ein Schmerzreiz, der in ACQ zeitlich mit dem Erscheinen grüner Quadrate (CS+) gekoppelt wird. Bei blauen Quadraten (CS-) sowie in beiden Extinktionsphasen erfolgt kein Schmerzreiz. Anhand der Ausprägung des rs1997794-Polymorphismus werden drei Gruppen gebildet (TT vs. CT vs. CC) und in der fMRT sowie Hautleitfähigkeitsmessung miteinander verglichen. PDYN-Effekte in der fMRT werden mit SPM8 und einer multiplen Regressionsanalyse untersucht (Regions of interest, ROIs: Amygdala und Ventromedialer Präfrontaler Cortex (VMPFC)). PDYN-Effekte in der Hautleitfähigkeitsmessung werden in SPSS 22 mit einer Varianzanalyse (ANOVA) mit Messwiederholungen analysiert. Ergebnisse: In der fMRT-Auswertung mittels multipler Regressionsanalyse können keine PDYN-Effekte auf die Amygdala-Aktivierung in den Extinktionsphasen festgestellt werden. Allerdings zeigt sich ein PDYN-Effekt auf die VMPFC-Aktivität in EX2. Die exponentiell ansteigende Aktivität des VMPFC wird während CS+ (GrünExpo EX2) mit steigender Anzahl an T-Allelen (TT > CT > CC) verstärkt. Darüber hinaus korreliert die VMPFC-Aktivität bei GrünExpo EX2 signifikant mit dem Summenscore des State-Trait-Anxiety Inventory-S (STAI-S; Fragebogen für Ängstlichkeit) nach EX2. In keiner der Subtasks zeigt sich ein PDYN-Effekt auf die Hautleitfähigkeitsmessung. Schlussfolgerung: Die Ergebnisse von Bilkei-Gorzo et al. (2012) können nicht repliziert werden. Spekuliert werden darf allerdings, ob T-Allelträger generell eine Prädisposition zu verstärkter Amygdala-Aktivität bei CS+ besitzen, die sich jedoch nur bei der TT-Subgruppe der Vorgängerstudie manifestiert. In der vorliegenden Replikationsstudie könnte eine vermehrte Amygdala-Aktivität durch den VMPFC supprimiert werden, was sowohl die fehlenden Geneffekte auf die Amygdala-Aktivität als auch den Zusammenhang zwischen VMPFC-Aktivität und T-Allelen zu erklären vermag. Natürlich lässt sich die Nicht-Replizierbarkeit des Geneffektes auch als allgemeine Problematik von Imaging-Genetics-Studien (geringe Einzelgen-Effektstärken und kleine Stichproben) deuten. Zur weiteren Investigation sind Studien mit größeren Stichproben notwendig.Abstract Background: Experimental studies from Bilkei-Gorzo et al. (2012) suggest a major impact of prodynorphine (PDYN) on acquisition and extinction of pain-induced behavior. In humans, a corresponding single-nucleotid-polymorphism can be detected at rs1997794 (PDYN-promoter-region). T-allele-homozygotes at rs1997794 show increased pain-induced fear acquisition and extinction capabilities compared to C-allele-carriers. The objective of this study is to replicate the human experimental part of the precursor study from Bilkei-Gorzo et al. (2012) in a larger sample and further, to analyze PDYN-effects on functional Magnetic Resonance Imaging (fMRI) and Skin Conductance Responses (SCR). Methods: A sample of 104 healthy adults (60 women; mean age: 28,17 years) is to be examined using the fMRI-paradigm from Bilkei-Gorzo et al. (2012): The experimental set-up consists of acquisition (ACQ), early (EX1) and late extinction (EX2). A painful stimulus serves as unconditioned stimulus (UCS), timed with the appearance of green squares (CS+) during ACQ. During the extinction phases and during the presentation of blue squares (CS-) no painful stimulus is applied. On the basis of rs1997794-characteristics three groups (TT vs. CT vs. CC) are established and compared using fMRI and SCR. PDYN-effects in fMRI are investigated with SPM8 and multiple regressions analyses (regions of interest (ROIs): amygdala and ventromedial prefrontal cortex (VMPFC)). PDYN-effects in SCR are analysed in SPSS 22 with an analyses of variance (ANOVA) with multiple measures. Results: According to the evaluation of fMRI-results of multiple regression analyses no PDYN-effects on amygdala activity in extinction phases can be detected. Nevertheless, a PDYN-effect on VMPFC activity during EX2 becomes apparent. The exponentially increasing VMPFC activity at the time of CS+ (green expo EX2) is enhanced with an increasing number of T-alleles (TT > CT > CC). Furthermore, VMPFC activity in green expo EX2 correlates significantly with the State-Trait-Anxiety Inventory-S (STAI-S)-score after-EX2. A PDYN-effect on Skin Conductance Responses cannot be demonstrated in any subtask. Conclusion: The results from Bilkei-Gorzo A. et al. (2012) cannot be replicated. Nonetheless, allowing for speculation, T-allele-carriers might bear a general predisposition to higher amygdala activity at the time of CS+, manifesting itself in the TT-subgroup only. In the present replication study an increased amygdala activity might be suppressed by the VMPFC, explaining the lack of PDYN-effects on amygdala activity and the correlation between VMPFC activity and T-alleles. However, the non-replicability of the PDYN-effects could also be interpreted as a general problem in imaging genetics studies (low single-gene-power and small samples). Large-sample-studies are required for further investigation

Surviving in Europe : geopolitics of biodiversity conservation illustrated by a proxy species Viola uliginosa

Building strategies for continental-scale conservation is challenging due to evolutionary and geopolitical problems. How do policy choices arise from this setting? In this study, we integrate ecological research with policy analysis to examine the problem field with a case study research. We use a violet species endemic to Europe, Viola uliginosa, as a proxy for a significant European Union (EU)-Russian biodiversity pattern and its conservation. The violet's core populations locate in Belarus, Ukraine, and Russia, and all populations in the EU are peripheral. The species is endangered in 12 EU member states and in decline in many places elsewhere. To analyze the choices of conservation, we gathered data on its ecology, distribution, and conservation mechanisms across Europe, putting additional emphasis on the EU enlargement and long-term site histories in Finland. We found that the survival of the species in the EU depends on the enlargement negotiations, conflicts between the EU biodiversity and agricultural policies, selection of the species to national Red Lists and the Habitats Directive, and contingent site histories depending on the conservation activities by civic actors and the member states. While the evolutionary aspect emphasizes the genetic differentiation potential of peripheral populations, the geopolitical aspect characterizes the EU as simultaneous spaces of a monotopia, borderlands, and polycentric development. We conclude that intersections between these geopolitical spaces can be used with evolutionary perspectives to identify local, European, and network-driven policy choices of conservation.Peer reviewe

Разработка технологии изготовления детали «Ось»

Цель работы – Согласно приведенным чертежам и годовой программой выпускай, разработать технологии изготовления деталей "ось привода". В работе дается обоснование выполнения ВКР и опытно-конструкторских работ, анализируется чертеж детали и ее технологичность, определяется вид изготовления, принцип выбора заготовки в соответствии с ее материалом и серийным выпуском. описано производство, составлен чертеж заготовки, маршрут обработки детали с представлением рабочих эскизов и описанием переходов для каждой операции, рассчитаны припуски на обработку и технологические размеры, рассчитывался режимы резания для каждого технологического перехода и требуемая мощность оборудования, задавалась модель станка и рассчитывалось время выполнения каждой операции.Purpose of work - release and develop manufacturing technology based on drawings and annual plans. The paper provides a justification for the implementation of research and development work and experimental design work, analyzes the drawing of the part and its manufacturability, determines the type of manufacture, the principle of selecting the workpiece in accordance with its material and serial production. a drawing of the workpiece was drawn up, the processing route of the part with the presentation of working sketches and a description of the transitions for each operation, the cutting conditions were calculated for each technological transition and the required power of the equipment, the model of the machine was set and the time of each operation was calculated

Glucose transporter-1 deficiency syndrome: the expanding clinical and genetic spectrum of a treatable disorder

Glucose transporter-1 deficiency syndrome is caused by mutations in the SLC2A1 gene in the majority of patients and results in impaired glucose transport into the brain. From 2004-2008, 132 requests for mutational analysis of the SLC2A1 gene were studied by automated Sanger sequencing and multiplex ligation-dependent probe amplification. Mutations in the SLC2A1 gene were detected in 54 patients (41%) and subsequently in three clinically affected family members. In these 57 patients we identified 49 different mutations, including six multiple exon deletions, six known mutations and 37 novel mutations (13 missense, five nonsense, 13 frame shift, four splice site and two translation initiation mutations). Clinical data were retrospectively collected from referring physicians by means of a questionnaire. Three different phenotypes were recognized: (i) the classical phenotype (84%), subdivided into early-onset (<2 years) (65%) and late-onset (18%); (ii) a non-classical phenotype, with mental retardation and movement disorder, without epilepsy (15%); and (iii) one adult case of glucose transporter-1 deficiency syndrome with minimal symptoms. Recognizing glucose transporter-1 deficiency syndrome is important, since a ketogenic diet was effective in most of the patients with epilepsy (86%) and also reduced movement disorders in 48% of the patients with a classical phenotype and 71% of the patients with a non-classical phenotype. The average delay in diagnosing classical glucose transporter-1 deficiency syndrome was 6.6 years (range 1 month-16 years). Cerebrospinal fluid glucose was below 2.5 mmol/l (range 0.9-2.4 mmol/l) in all patients and cerebrospinal fluid : blood glucose ratio was below 0.50 in all but one patient (range 0.19-0.52). Cerebrospinal fluid lactate was low to normal in all patients. Our relatively large series of 57 patients with glucose transporter-1 deficiency syndrome allowed us to identify correlations between genotype, phenotype and biochemical data. Type of mutation was related to the severity of mental retardation and the presence of complex movement disorders. Cerebrospinal fluid : blood glucose ratio was related to type of mutation and phenotype. In conclusion, a substantial number of the patients with glucose transporter-1 deficiency syndrome do not have epilepsy. Our study demonstrates that a lumbar puncture provides the diagnostic clue to glucose transporter-1 deficiency syndrome and can thereby dramatically reduce diagnostic delay to allow early start of the ketogenic die

Full-length Isoform Sequencing for Resolving the Molecular Basis of Charcot-Marie-Tooth 2A.

OBJECTIVES: Transcript sequencing of patient-derived samples has been shown to improve the diagnostic yield for solving cases of suspected Mendelian conditions, yet the added benefit of full-length long-read transcript sequencing is largely unexplored. METHODS: We applied short-read and full-length transcript sequencing and mitochondrial functional studies to a patient-derived fibroblast cell line from an individual with neuropathy that previously lacked a molecular diagnosis. RESULTS: We identified an intronic homozygous MFN2 c.600-31T&gt;G variant that disrupts the branch point critical for intron 6 splicing. Full-length long-read isoform complementary DNA (cDNA) sequencing after treatment with a nonsense-mediated mRNA decay (NMD) inhibitor revealed that this variant creates 5 distinct altered splicing transcripts. All 5 altered splicing transcripts have disrupted open reading frames and are subject to NMD. Furthermore, a patient-derived fibroblast line demonstrated abnormal lipid droplet formation, consistent with MFN2 dysfunction. Although correctly spliced full-length MFN2 transcripts are still produced, this branch point variant results in deficient MFN2 levels and autosomal recessive Charcot-Marie-Tooth disease, axonal, type 2A (CMT2A). DISCUSSION: This case highlights the utility of full-length isoform sequencing for characterizing the molecular mechanism of undiagnosed rare diseases and expands our understanding of the genetic basis for CMT2A

Dominant-negative variant in SLC1A4 causes an autosomal dominant epilepsy syndrome.

SLC1A4 is a trimeric neutral amino acid transporter essential for shuttling L-serine from astrocytes into neurons. Individuals with biallelic variants in SLC1A4 are known to have spastic tetraplegia, thin corpus callosum, and progressive microcephaly (SPATCCM) syndrome, but individuals with heterozygous variants are not thought to have disease. We identify an 8-year-old patient with global developmental delay, spasticity, epilepsy, and microcephaly who has a de novo heterozygous three amino acid duplication in SLC1A4 (L86_M88dup). We demonstrate that L86_M88dup causes a dominant-negative N-glycosylation defect of SLC1A4, which in turn reduces the plasma membrane localization of SLC1A4 and the transport rate of SLC1A4 for L-serine

Clozapine for Treatment-Resistant Bipolar Disorder: A Systematic Review

Objective To evaluate the efficacy and safety of clozapine for treatment-resistant bipolar disorder (TRBD). Methods A systematic review of randomized controlled studies, open-label prospective studies, and retrospective studies of patients with TRBD was carried out. Interventions included clozapine monotherapy or clozapine combined with other medications. Outcome measures were efficacy and adverse drug reactions (ADRs). Results Fifteen clinical trials with a total sample of 1,044 patients met the inclusion criteria. Clozapine monotherapy or clozapine combined with other treatments for TRBD was associated with improvement in: (i) symptoms of mania, depression, rapid cycling, and psychotic symptoms, with many patients with TRBD achieving a remission or response; (ii) the number and duration of hospitalizations, the number of psychotropic co-medications, and the number of hospital visits for somatic reasons for intentional self-harm/overdose; (iii) suicidal ideation and aggressive behavior; and (iv) social functioning. In addition, patients with TRBD showed greater clinical improvement in long-term follow-up when compared with published schizophrenia data. Sedation (12%), constipation (5.0%), sialorrhea (5.2%), weight gain (4%), and body ache/pain (2%) were the commonly reported ADRs; however, these symptoms but did not usually require drug discontinuation. The percentage of severe ADRs reported, such as leukopenia (2%), agranulocytosis (0.3%), and seizure (0.5%), appeared to be lower than those reported in the published schizophrenia literature. Conclusion The limited current evidence supports the concept that clozapine may be both an effective and a relatively safe medication for TRBD

Developmental epileptic encephalopathy in &lt;i&gt;DLG4&lt;/i&gt;-related synaptopathy

Objective: The postsynaptic density protein of excitatory neurons PSD-95 is encoded by discs large MAGUK scaffold protein 4 (DLG4), de novo pathogenic variants of which lead to DLG4-related synaptopathy. The major clinical features are developmental delay, intellectual disability (ID), hypotonia, sleep disturbances, movement disorders, and epilepsy. Even though epilepsy is present in 50% of the individuals, it has not been investigated in detail. We describe here the phenotypic spectrum of epilepsy and associated comorbidities in patients with DLG4-related synaptopathy. Methods: We included 35 individuals with a DLG4 variant and epilepsy as part of a multicenter study. The DLG4 variants were detected by the referring laboratories. The degree of ID, hypotonia, developmental delay, and motor disturbances were evaluated by the referring clinician. Data on awake and sleep electroencephalography (EEG) and/or video-polygraphy and brain magnetic resonance imaging were collected. Antiseizure medication response was retrospectively assessed by the referring clinician. Results: A large variety of seizure types was reported, although focal seizures were the most common. Encephalopathy related to status epilepticus during slow-wave sleep (ESES)/developmental epileptic encephalopathy with spike-wave activation during sleep (DEE-SWAS) was diagnosed in &gt;25% of the individuals. All but one individual presented with neurodevelopmental delay. Regression in verbal and/or motor domains was observed in all individuals who suffered from ESES/DEE-SWAS, as well as some who did not. We could not identify a clear genotype-phenotype relationship even between individuals with the same DLG4 variants. Significance: Our study shows that a subgroup of individuals with DLG4-related synaptopathy have DEE, and approximately one fourth of them have ESES/DEE-SWAS. Our study confirms DEE as part of the DLG4-related phenotypic spectrum. Occurrence of ESES/DEE-SWAS in DLG4-related synaptopathy requires proper investigation with sleep EEG

High incidence of Noonan syndrome features including short stature and pulmonic stenosis in patients carrying NF1 missense mutations affecting p.Arg1809: genotype-phenotype correlation

Neurofibromatosis type 1 (NF1) is one of the most frequent genetic disorders, affecting 1:3,000 worldwide. Identification of genotype-phenotype correlations is challenging because of the wide range clinical variability, the progressive nature of the disorder, and extreme diversity of the mutational spectrum. We report 136 individuals with a distinct phenotype carrying one of five different NF1 missense mutations affecting p.Arg1809. Patients presented with multiple cafe-au-lait macules (CALM) with or without freckling and Lisch nodules, but no externally visible plexiform neurofibromas or clear cutaneous neurofibromas were found. About 25% of the individuals had Noonan-like features. Pulmonic stenosis and short stature were significantly more prevalent compared with classic cohorts (P<0.0001). Developmental delays and/or learning disabilities were reported in over 50% of patients. Melanocytes cultured from a CALM in a segmental NF1-patient showed two different somatic NF1 mutations, p.Arg1809Cys and a multi-exon deletion, providing genetic evidence that p.Arg1809Cys is a loss-of-function mutation in the melanocytes and causes a pigmentary phenotype. Constitutional missense mutations at p.Arg1809 affect 1.23% of unrelated NF1 probands in the UAB cohort, therefore this specific NF1 genotype-phenotype correlation will affect counseling and management of a significant number of patients