Ultrafast Delamination of Graphite into High-Quality Graphene Using Alternating Currents

To bridge the gap between laboratory‐scale studies and commercial applications, mass production of high quality graphene is essential. A scalable exfoliation strategy towards the production of graphene sheets is presented that has excellent yield (ca. 75 %, 1–3 layers), low defect density (a C/O ratio of 21.2), great solution‐processability, and outstanding electronic properties (a hole mobility of 430 cm2 V−1 s−1). By applying alternating currents, dual exfoliation at both graphite electrodes enables a high production rate exceeding 20 g h−1 in laboratory tests. As a cathode material for lithium storage, graphene‐wrapped LiFePO4 particles deliver a high capacity of 167 mAh g−1 at 1 C rate after 500 cycles

Protocol of a prospective study on the diagnostic value of transcranial duplex scanning of the substantia nigra in patients with parkinsonian symptoms

<p>Abstract</p> <p>Background</p> <p>Parkinson's disease (PD) is the second most common neurodegenerative disorder. As there is no definitive diagnostic test, its diagnosis is based on clinical criteria. Recently transcranial duplex scanning (TCD) of the substantia nigra in the brainstem has been proposed as an instrument to diagnose PD. We and others have found that TCD scanning of substantia nigra duplex is a relatively accurate diagnostic instrument in patients with parkinsonian symptoms. However, all studies on TCD so far have involved well-defined, later-stage PD patients, which will obviously lead to an overestimate of the diagnostic accuracy of TCD.</p> <p>We have therefore set out to conduct a prospective study testing the diagnostic accuracy of TCD in patients with a parkinsonism of unclear origin.</p> <p>Methods/Design</p> <p>We will enrol 250 consecutive patients, who are referred to neurology outpatient clinics of two teaching hospitals, for analysis of clinically unclear parkinsonism. Patients, whose parkinsonism is clearly diagnosable at the first visit, will be excluded from the study. All patients will undergo a TCD of the substantia nigra. As a surrogate gold standard we will use the consensus clinical diagnosis reached by two independent, blinded, movement disorder specialist neurologists after 2 years follow-up. At the time of TCD, patients will also undergo a SPECT scan of the brain.</p> <p>Discussion</p> <p>As this prospective trial enrols only patients with an early-stage parkinsonism, it will yield data on the diagnostic accuracy of TCD that is relevant to daily clinical practice: The neurologist needs a diagnostic tool that provides additional information in patients with a clinically indefinable parkinsonian syndrome. The above described observational longitudinal study was designed to explicitly study this aspect in the diagnostic process.</p> <p>Trial registration</p> <p><b>(ITRSCC) NCT00368199</b></p

Human Bone Marrow Mesenchymal Stem Cells Display Anti-Cancer Activity in SCID Mice Bearing Disseminated Non-Hodgkin's Lymphoma Xenografts

Abstract BACKGROUND: Although multimodality treatment can induce high rate of remission in many subtypes of non-Hodgkin's lymphoma (NHL), significant proportions of patients relapse with incurable disease. The effect of human bone marrow (BM) mesenchymal stem cells (MSC) on tumor cell growth is controversial, and no specific information is available on the effect of BM-MSC on NHL. METHODOLOGY/PRINCIPAL FINDINGS: The effect of BM-MSC was analyzed in two in vivo models of disseminated non-Hodgkin's lymphomas with an indolent (EBV(-) Burkitt-type BJAB, median survival = 46 days) and an aggressive (EBV(+) B lymphoblastoid SKW6.4, median survival = 27 days) behavior in nude-SCID mice. Intra-peritoneal (i.p.) injection of MSC (4 days after i.p. injection of lymphoma cells) significantly increased the overall survival at an optimal MSC:lymphoma ratio of 1:10 in both xenograft models (BJAB+MSC, median survival = 58.5 days; SKW6.4+MSC, median survival = 40 days). Upon MSC injection, i.p. tumor masses developed more slowly and, at the histopathological observation, exhibited a massive stromal infiltration coupled to extensive intra-tumor necrosis. In in vitro experiments, we found that: i) MSC/lymphoma co-cultures modestly affected lymphoma cell survival and were characterized by increased release of pro-angiogenic cytokines with respect to the MSC, or lymphoma, cultures; ii) MSC induce the migration of endothelial cells in transwell assays, but promoted endothelial cell apoptosis in direct MSC/endothelial cell co-cultures. CONCLUSIONS/SIGNIFICANCE: Our data demonstrate that BM-MSC exhibit anti-lymphoma activity in two distinct xenograft SCID mouse models of disseminated NHL

Analysis of shared heritability in common disorders of the brain

ience, this issue p. eaap8757 Structured Abstract INTRODUCTION Brain disorders may exhibit shared symptoms and substantial epidemiological comorbidity, inciting debate about their etiologic overlap. However, detailed study of phenotypes with different ages of onset, severity, and presentation poses a considerable challenge. Recently developed heritability methods allow us to accurately measure correlation of genome-wide common variant risk between two phenotypes from pools of different individuals and assess how connected they, or at least their genetic risks, are on the genomic level. We used genome-wide association data for 265,218 patients and 784,643 control participants, as well as 17 phenotypes from a total of 1,191,588 individuals, to quantify the degree of overlap for genetic risk factors of 25 common brain disorders. RATIONALE Over the past century, the classification of brain disorders has evolved to reflect the medical and scientific communities' assessments of the presumed root causes of clinical phenomena such as behavioral change, loss of motor function, or alterations of consciousness. Directly observable phenomena (such as the presence of emboli, protein tangles, or unusual electrical activity patterns) generally define and separate neurological disorders from psychiatric disorders. Understanding the genetic underpinnings and categorical distinctions for brain disorders and related phenotypes may inform the search for their biological mechanisms. RESULTS Common variant risk for psychiatric disorders was shown to correlate significantly, especially among attention deficit hyperactivity disorder (ADHD), bipolar disorder, major depressive disorder (MDD), and schizophrenia. By contrast, neurological disorders appear more distinct from one another and from the psychiatric disorders, except for migraine, which was significantly correlated to ADHD, MDD, and Tourette syndrome. We demonstrate that, in the general population, the personality trait neuroticism is significantly correlated with almost every psychiatric disorder and migraine. We also identify significant genetic sharing between disorders and early life cognitive measures (e.g., years of education and college attainment) in the general population, demonstrating positive correlation with several psychiatric disorders (e.g., anorexia nervosa and bipolar disorder) and negative correlation with several neurological phenotypes (e.g., Alzheimer's disease and ischemic stroke), even though the latter are considered to result from specific processes that occur later in life. Extensive simulations were also performed to inform how statistical power, diagnostic misclassification, and phenotypic heterogeneity influence genetic correlations. CONCLUSION The high degree of genetic correlation among many of the psychiatric disorders adds further evidence that their current clinical boundaries do not reflect distinct underlying pathogenic processes, at least on the genetic level. This suggests a deeply interconnected nature for psychiatric disorders, in contrast to neurological disorders, and underscores the need to refine psychiatric diagnostics. Genetically informed analyses may provide important "scaffolding" to support such restructuring of psychiatric nosology, which likely requires incorporating many levels of information. By contrast, we find limited evidence for widespread common genetic risk sharing among neurological disorders or across neurological and psychiatric disorders. We show that both psychiatric and neurological disorders have robust correlations with cognitive and personality measures. Further study is needed to evaluate whether overlapping genetic contributions to psychiatric pathology may influence treatment choices. Ultimately, such developments may pave the way toward reduced heterogeneity and improved diagnosis and treatment of psychiatric disorders

Analysis of 30 Putative <em>BRCA1</em> Splicing Mutations in Hereditary Breast and Ovarian Cancer Families Identifies Exonic Splice Site Mutations That Escape <em>In Silico</em> Prediction

<div><p>Screening for pathogenic mutations in breast and ovarian cancer genes such as <em>BRCA1/2</em>, <em>CHEK2</em> and <em>RAD51C</em> is common practice for individuals from high-risk families. However, test results may be ambiguous due to the presence of unclassified variants (UCV) in the concurrent absence of clearly cancer-predisposing mutations. Especially the presence of intronic or exonic variants within these genes that possibly affect proper pre-mRNA processing poses a challenge as their functional implications are not immediately apparent. Therefore, it appears necessary to characterize potential splicing UCV and to develop appropriate classification tools. We investigated 30 distinct <em>BRCA1</em> variants, both intronic and exonic, regarding their spliceogenic potential by commonly used <em>in silico</em> prediction algorithms (HSF, MaxEntScan) along with <em>in vitro</em> transcript analyses. A total of 25 variants were identified spliceogenic, either causing/enhancing exon skipping or activation of cryptic splice sites, or both. Except from a single intronic variant causing minor effects on <em>BRCA1</em> pre-mRNA processing in our analyses, 23 out of 24 intronic variants were correctly predicted by MaxEntScan, while HSF was less accurate in this cohort. Among the 6 exonic variants analyzed, 4 severely impair correct pre-mRNA processing, while the remaining two have partial effects. In contrast to the intronic alterations investigated, only half of the spliceogenic exonic variants were correctly predicted by HSF and/or MaxEntScan. These data support the idea that exonic splicing mutations are commonly disease-causing and concurrently prone to escape <em>in silico</em> prediction, hence necessitating experimental <em>in vitro</em> splicing analysis.</p> </div

RT-PCR analyses of <i>BRCA1</i> exon 19 and flanking sequences.

<p><b><i>A</i></b><i>)</i> Compared with controls C (1) to C (5), the variants <i>IVS18-2delA</i> and <i>IVS19+2T>G</i> elevate exclusion of exon 19 (Δex19). Regarding the variant <i>IVS18-2delA</i>, two mRNA samples derived from two unrelated mutation carriers were analyzed. Effects of <i>IVS18-6C>A</i> on <i>BRCA1</i> pre-mRNA processing were not observed. <i>IVS19+1delG</i> did not associate with a suspicious splicing pattern as shown by RT-PCR followed by gel electrophoresis. * Note that <i>IVS19+1delG</i> causes a 1 nt deletion on transcript level not detectable by agarose gel electrophoresis. <b><i>B</i></b><i>)</i> Direct sequencing of <i>IVS19+1delG</i> samples following RT-PCR revealed the deletion of the last nucleotide of exon 19 on mRNA level due to the activation of a cryptic splice site, which incorporates the last nucleotide of exon 19. NTC = no template control.</p

Classification of putative <i>BRCA1</i> splicing mutations.

<p><b>A:</b> Variants that severely affect splicing, <b>B:</b> Variants having a partial effect only, and <b>C:</b> Variants that do not affect processing of <i>BRCA1</i> pre-mRNA species in PBL.</p>*<p>Variants with severe impact on splicing are considered as likely pathogenic (class 4) according to the classification system proposed by Plon et al., <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0050800#pone.0050800-Plon1" target="_blank">[42]</a>, while variants with only partial effects on splicing remain of uncertain clinical significance (class 3). Variant descriptions (BIC nomenclature, HGVS nomenclature), consequences on transcript- and protein levels, family IDs, analyzed index patients (phenotypes, age at onset), family histories (phenotypes, age at onset) and ethnic backgrounds are given. Family members carrying the same <i>BRCA1</i> variant are indicated (asterisk). All other listed family members were not available for analysis. Abbreviation: BC = breast cancer; OC = ovarian cancer; n.a. = not affected; bil = bilateral; ProC = prostate carcinoma; MTX = mastectomy; DCIS = Ductal carcinoma <i>in situ</i>.</p

RT-PCR analyses of <i>BRCA1</i> exons 5 (A, B), 9 (C, D), and flanking sequences.

<p><b><i>A</i></b><i>)</i> Compared with controls C (1) to C (5), the variants <i>IVS4-18T>G</i> and <i>IVS4-1G>C</i> elevate exon 5 exclusion (Δex5), while <i>IVS5+1G>T</i>, <i>IVS5+1G>C</i> and <i>IVS5+1G>A</i> promote the usage of an upstream cryptic splice site, resulting in a 22 bp deletion on mRNA level (Δ22nt ex5). Regarding the variant <i>IVS5+1G>T</i>, two mRNA samples derived from two related mutation carriers were analyzed. NTC = no template control. Effects of the variant <i>IVS5+23T>A</i> on <i>BRCA1</i> pre-mRNA processing were not observed. <b><i>B</i></b><i>)</i> Compared with two control samples, enhanced exon 5 skipping in <i>IVS4-18T>G</i> and <i>IVS4-1G>C</i> samples was confirmed by quantitative real-time RT-PCR analyses. Expression data are given as mean ± standard deviation (s.d.). Relative to an internal <i>BRCA1</i> control set to 100% (amplicon spanning exon 6 and 7 sequences, <i>BRCA1</i> ex6/7), the relative amounts of transcripts lacking exon 5 (<i>BRCA1</i> ex2/3/6) account for 3.49% (+1.01, −0.78) and 3.03% (+1.11, −0.81) in control samples, respectively, while the relative amounts of <i>BRCA1</i> ex2/3/6 transcripts are approximately 3fold increased in <i>IVS4-18T>G</i> samples (10.83%, +0.76, −0.71). <i>IVS4-1G>C</i> increases the relative amount of <i>BRCA1</i> ex2/3/6 transcripts to 56.21% (+13.77, −11.06), while the share of transcripts harbouring exon 5 sequences is significantly reduced. Three levels of statistical significance were discriminated: * = P<0.05, ** = P<0.01, *** = P<0.001 (t-test). <b><i>C</i></b><i>)</i> The variant <i>710C>T,C197C</i> elevates skipping of exon 9 (Δex9) compared with controls. Total mRNA samples derived from two unrelated <i>710C>T, C197C</i> mutation carriers were analyzed. <b><i>D</i></b><i>)</i> Enhanced exon 9 skipping was confirmed by quantitative real-time analysis. While transcripts lacking exon 9 (<i>BRCA1</i> ex8/10) account for 2.51% (+0.23, −0.21) and 2.14% (+0.35, −0.30) relative to the respective internal controls, the amounts of <i>BRCA1</i> ex8/10 mRNA species are approximately 2fold increased in samples derived from two independent patients carrying the <i>710C>T, C197C</i> variant (5.23%, +0.70, −0.62; 6.92%, +0.55, −0.51). Similar results were observed when analyzing the relative amounts of transcripts lacking exons 9 and 10 (<i>BRCA1</i> ex8/11). In controls, relative <i>BRCA1</i> ex8/11 levels account for 7.69% (+0.70, −0.64) and 8.04% (+1.30, −1.12) and 16.73% (+2.25, −1.98) and 15.48% (+1.23, −1.14) in samples derived from two independent <i>710C>T, C197C</i> carriers.</p