A new standard nomenclature for proteins related to Apx and Shroom

Shroom is a recently-described regulator of cell shape changes in the developing nervous system. This protein is a member of a small family of related proteins that are defined by sequence similarity and in most cases by some link to the actin cytoskeleton. At present these proteins are named Shroom, APX, APXL, and KIAA1202. In light of the growing interest in this family of proteins, we propose here a new standard nomenclature

Karakteristik Nanoemulsi Minyak Sawit Merah Yang Diperkaya Beta Karoten

Red palm oil (RPO) and β-carotene are insoluble in water. It makescan be used to improve RPO and βThis research is aimed to produce stable RPO nanoemulsion enriched withβ-carotene. The research was conducted in the SEAFAST CENTERLaboratory, Bogor Agriculture University from January to Septemberfollowing steps, i.e. enrichment of RPO with βusing a high pressure homogenizer at a pressure of 34.5 MPa in 10 cycles.The ratio of RPO and water in the mixture were 5 : 95; 7.5 : 92.5; and 10 :10% (w/w) of the total emulsions. In the second stage, nanoemulsionswere prepared on various RPO percentage of 2, 4, and 6% (w/w) andhad a droplet size from 115.1 to 145.2 nm and stable. Nanoemulsions wereresulting from the second stage had droplet size from 94.9 to 125.5 nm,and β-carotene content were 47.6 to 130.9 mg/l. Droplet size ofnanoemulsions is less than 125 nm. It can be produced with RPO an

Pembuatan Ethanol Dari Jerami Padi Dengan Proses Hidrolisis Dan Fermentasi

Jerami Padi banyak mengandung Pati, Selulosa dan Glukosa yang cukup tinggi. Alkohol dapat dihasilkan dari tanaman yang banyak mengandung senyawa selulosa dengan menggunakan bantuan aktivitas mikroba salah satu jenis tanamannya adalah jerami padi Tujuan penelitian ini yaitu untuk mendapatkan kadar ethanol yang terbaik pada jerami padi dengan menggunakan proses hidrolisis dan fermentasi. Kondisi yang ditetapkan larutan Hidrolisis sebanyak 2500 ml, pH hidrolisis 3, waktu hidrolisis 2 hari, dan pH fermentasi sebesar 4,5, sedangkan peubah yang dijalankan adalah waktu fermentasi (2,3,4,5,6,7 (hari)), berat jerami padi (40,50,60 (gram)), dan volume stater yang ditambahkan (8%, 10%, 12%, kali volume cairan fermentasi). Hasil penelitian menunjukkan bahwa kondisi terbaik pada berat jerami 50 gram dengan volume stater yang ditambahkan sebanyak 12% volume cairan fermentasi yang difermentasi selama 7 hari yang menghasilkan kadar ethanol sebesar 12,89%. Jerami padi dapat digunakan sebagai bahan baku alternatif pembuatan bioethanol

Automated High-Throughput Characterization of Single Neurons by Means of Simplified Spiking Models

Single-neuron models are useful not only for studying the emergent properties of neural circuits in large-scale simulations, but also for extracting and summarizing in a principled way the information contained in electrophysiological recordings. Here we demonstrate that, using a convex optimization procedure we previously introduced, a Generalized Integrate-and-Fire model can be accurately fitted with a limited amount of data. The model is capable of predicting both the spiking activity and the subthreshold dynamics of different cell types, and can be used for online characterization of neuronal properties. A protocol is proposed that, combined with emergent technologies for automatic patch-clamp recordings, permits automated, in vitro high-throughput characterization of single neurons

Factorial validation of the Attitudes toward Women Scale for Adolescents (AWSA) in assessing sexual behaviour patterns in Bolivian and Ecuadorian adolescents

Background: Adolescents' health is greatly influenced by social determinants, including gender norms. Although research has shown that there is an association between gender attitudes and adolescents' sexual behaviour, few studies have assessed this relationship carefully. The Attitudes toward Women Scale for Adolescents (AWSA) is widely used to assess gender attitudes among adolescents; however, to our knowledge it has not been applied in Latin America. Objective: To apply AWSA in Latin America for the first time, to perform a factorial validation of this scale and to assess the relationship of gender attitudes and sexual behaviour in Bolivian and Ecuadorian adolescents. Design: This cross-sectional study was carried out in 2011 among 14-18 year olds in 20 high schools in Cochabamba (Bolivia) and six in Cuenca (Ecuador) as a part of a larger project. Schools were purposively selected. A Spanish version of the 12-item AWSA was employed for this study. The assessed aspects of adolescent sexual behaviour were: reported sexual intercourse, reported positive experience during last sexual intercourse and reported current use of contraception. The psychometric properties of AWSA were investigated, and both explanatory and confirmatory factorial analyses were performed. Results: The number of questionnaires included in the analysis was 3,518 in Bolivia and 2,401 in Ecuador. A factorial analysis of AWSA resulted in three factors: power dimension (PD), equality dimension (ED) and behavioural dimension (BD). ED showed the highest correlates with adolescent sexual behaviour. Higher scores of this dimension were associated with a more positive experience of sexual relationships, a higher current use of modern contraception and greater sexual activity among girls. Conclusions: This study revealed a three-factorial structure of AWSA and demonstrated that by employing factors, the sensitivity of AWSA increases as compared to using the scale as a whole to assess sexual behaviour. This could have important implications for future research on gender and the sexual experiences of adolescents

Molekulargenetische Charakterisierung der beiden Gene, FBXO25 und KIAA1202, die durch Translokationsbruchpunkte in einer geistig behinderten Patientin unterbrochen wurden

Title Page Acknowledgement Table of Contents Index of Figures Index of Tables Acronymes & Abbreviations Summary Zusammenfassung 1\. Introduction 2\. Materials & Methods 3\. Results 4\. Discussion 5\. Outlook 6\. Epilogue - X-linked mental retardation. Is it really on the X? 7\. Commentary - Should we place less importance on intelligence? Appendix A - Intelligence quotient: definition, evaluation and problems as a measure of intelligence Appendix B - Molecular mechanism and pattern of X chromosome inactivation Appendix C - From the principle of heredity to the molecular elucidation of inborn errors of metabolism Appendix D - Mutations in human pathology Appendix E - Polymerase chain reaction primers Appendix F - Polymerase chain reaction cycling conditions Appendix G - Denaturing high-performance liquid chromatography conditions Appendix H - Gene, mRNA and protein symbols ReferencesTo date, mutations in ~75 different genes have been associated with mental retardation (MR), but these likely only represent the tip of the proverbial iceberg. A diminished ability to adapt to the daily demands of a normal social environment, an onset during childhood and an intelligence quotient < 70 define MR. Anatomical post-mortem examinations have demonstrated that dendritic spines, which are responsible for synaptic connections in the brain, are abnormal in different forms of MR. A large body of evidence shows that learning changes spine morphology and synaptic plasticity through remodelling of the Actin cytoskeleton. Mutations upsetting such remodelling cause MR. In uniting a pathologic phenotype with a specific genotype, disease-associated balanced chromosomal rearrangements provide a unique opportunity to identify genetic loci relevant to health. In this study, we characterised two novel genes that are disrupted by a de novo apparently balanced t(X;8) translocation in a mildly mentally retarded patient. While the autosomal breakpoint disrupts hFBXO25, the X-chromosomal breakpoint interrupts hKIAA1202. After characterising the genomic structure of hFBXO25 and its murine counterpart, establishing their expression patterns and investigating their subcellular localisation, we discovered that hFBXO25 is a bona fide F-box protein (FBP), which forms part of the SCFhFBXO25 complex. FBPs confer substrate specificity to the SCF E3 ubiquitin ligases, which poly-ubiquitinylate cell cycle regulators for proteasomal degradation. In line with their function, misregulation of and mutations in FBPs have been mostly associated with malignancies; a direct link between FBPs and MR has not yet been reported. Based on the genomic structure of hKIAA1202, and after having confirmed its expression in both foetal and adult human brain, we and several collaborators performed an extensive mutation analysis. This effort resulted in the identification of a second patient with mild MR carrying a different de novo apparently balanced translocation also disrupting hKIAA1202, a silent substitution possibly affecting an exonic splice enhancer in a small family with MR and a c.3266C>T missense exchange segregating with severe MR characteristic of the Stocco dos Santos syndrome. As hKIAA1202 is subject to X chromosome inactivation (XCI) and both translocation carriers show skewed XCI leading to inactivation of the intact X chromosome, their genetic make-up is predicted to represent a hKIAA1202 null mutation. However, RT-PCR experiments and generation of an a-hKIAA1202 antibody demonstrated that uncharacterised hKIAA1202 isoforms, which the c.3266C>T transition could affect, are still present in a cell line of the t(X;8) patient. Owing to secondary selection processes, certain mutations result in skewed XCI. Such mutations are often lethal to hemizygous male offspring, resulting in increased frequencies of spontaneous abortion in carrier females. Therefore, the Stocco dos Santos hKIAA1202 variant could explain the skewed XCI observed in all carrier women and may be the cause of the large number of spontaneous abortions and high rate of infant mortality in the family. Based on its domain structure, which is compatible with molecular scaffolding and binding of filamentous Actin (F-actin), we have established hKIAA1202 as one of the founding members of the Shroom family of cytoskeleton-associated proteins. By anchoring over-expressed hKIAA1202 to the mitochondria, we demonstrated that it can direct the subcellular distribution of F-actin in vivo. We also demonstrated co- localisation of endogenous hKIAA1202 with F-actin at sites of rapid Actin remodelling, such as the leading edge of fibroblasts and the neurites of differentiating neuronal cells. Preliminary in vitro studies indicate an interaction between hKIAA1202 and F-actin. Employing yeast two-hybrid methodology, we identified Vimentin, a cytoskeletal intermediate filament, and several proteins involved in chromatin remodelling and transcriptional regulation as putative hKIAA1202 interaction partners. Interference with transcriptional regulation, mainly through chromatin remodelling, has been established as a mechanism underlying the aetiology of MR. Taken together, we have provided evidence suggesting hKIAA1202 as a gene involved in human cognition. Such a role is unlikely for hFBXO25.Bis heute wurden Mutationen in ~75 unterschiedlichen Genen mit dem Krankheitsbild der geistigen Behinderung (GB) assoziiert. Es ist jedoch anzunehmen, dass weit mehr dieser Loci existieren. GB wird als eine verringerte Anpassungsfähigkeit an die Anforderungen definiert, die eine normale soziale Umgebung mit sich bringt, die in der Kindheit beginnt und mit einem Intelligenzquotienten von < 70 einhergeht. Durch Autopsien an GB- Patienten konnte gezeigt werden, dass dendritische Dornen (engl. spines), welche an der Bildung synaptischer Verbindungen im Gehirn verantwortlich sind, morphologische Veränderungen aufweisen. Es wird angenommen, dass das Lernen zu einem Umbau des Aktin-Zytoskeletts führt und somit die Morphologie der dendritischen Dornen und die synaptische Plastizität verändert. Mutationen, die eine Beeinträchtigung dieses Umbaus verursachen, können eine GB hervorrufen. Die Untersuchung von krankheitsassoziierten balancierten Chromosomenaberrationen stellt eine Möglichkeit dar, Phänotyp und Genotyp zu verknüpfen um neue, krankheitsrelevante Gene zu identifizieren. In dieser Arbeit wurden zwei neue Gene untersucht, die durch eine de novo balancierte t(X;8) Translokation in einer leicht geistig behinderten Patientin unterbrochen sind. Während der autosomale Bruchpunkt (BP) das hFBXO25 Gen unterbricht, trennt der X-chromosomale BP das hKIAA1202 Gen. Nach der Charakterisierung der genomischen Struktur des menschlichen hFBXO25 Gens und des homologen Gens der Maus, der Etablierung ihrer Expressionsmuster und der subzellulären Lokalisation konnte gezeigt werden, dass hFBXO25 ein F-box Protein (FBP) und Bestandteil des SCFhFBXO25 Komplexes ist. FBPe sind für die Substratspezifizität von SCF E3 Ubiquitinligasen verantwortlich, welche Regulatoren des Zellzyklus für den Abbau durch das Proteasom polyubiquitinylieren. Eine Fehlregulierung von FBPen und Mutationen in den entsprechenden Genen wurde bisher hauptsächlich mit Krebs assoziiert. Eine direkte Verbindung zwischen FBPen und GB wurde noch nicht beschrieben. Basierend auf der genomischen hKIAA1202 Struktur und nach Überprüfung seiner Expression in fötalem und adultem Gehirn, wurde in diesem Gen eine umfangreiche Mutationssuche durchgeführt. In einer kleinen Familie mit GB konnte ein stiller Basenaustausch nachgewiesen werden, welcher möglicherweise ein Spleiß-Enhancer Signal beeinträchtigt. In einer Familie mit Stocco dos Santos Syndrom wurde eine c.3266C>T Transition, die mit schwerer GB kosegregiert, nachgewiesen. Darüber hinaus wurde festgestellt, dass in einer leicht geistig behinderten Patientin mit einer de novo balancierten t(X;19) Translokation, das hKIAA1202 Gen unterbrochen wurde. Da das hKIAA1202 Gen der X-Inaktivierung (XI) unterliegt und beide Translokationsträgerinnen eine schiefe (engl. skewed) XI zeigen, wobei das intakte X Chromosom inaktiviert wurde, ist anzunehmen, dass ihre genetische Konstellation einen Verlust des hKIAA1202 Proteins widerspiegelt. Allerdings konnte die Expression von zuvor noch nicht bekannten hKIAA1202 RNA-Isoformen, die ebenfalls die c.3266C>T Transition aufweisen könnten, in einer Zellinie der t(X;8) Patientin durch RT- PCR Experimente nachgewiesen werden. Bei manchen Mutationen kommt es aufgrund von somatischer Selektion zu einer schiefen XI. In hemizygoten Nachkommen sind solche Mutationen häufig letal, wodurch bei Überträgerinnen in höherem Maße spontane Fehlgeburten vorkommen. Die Stocco dos Santos hKIAA1202 Variante kann demnach der Grund für die schiefe XI bei Überträgerinnen sein und für die häufigen Fehlgeburten und die hohe Säuglingssterblichkeit in der Familie eine Erklärung bieten. Basierend auf der Domänenstruktur des hKIAA1202 Proteins, die mit einer Funktion als molekulares Gerüst und mit der Bindung von Aktin kompatibel ist, wird hKIAA1202 als eines der Gründungsmitglieder der sogenannten Shroom Familie von zytoskelettassoziierten Proteinen etabliert. hKIAA1202 kann in vivo die subzelluläre Lokalisation von F-Aktin ändern, wenn es an die Mitochondrien verankert wird. Es konnte gezeigt werden, dass endogenes hKIAA1202 an Stellen schnellen Aktinumbaus, wie Führungslamellen in Fibroblasten und Neuriten von differenzierenden neuronalen Zellinien, mit F-Aktin kolokalisiert. Vorläufige in vitro Experimente weisen auf eine Interaktion zwischen hKIAA1202 und F-Aktin hin. Mittels des Hefe- Zweihybridsystems konnten mögliche hKIAA1202 Bindungspartner identifiziert werden. Dazu gehören Vimentin, ein intermediäres Filament, das eine Komponente des Zytoskeletts darstellt, sowie mehrere Proteine, die eine Rolle in der Kontrolle des Chromatinumbaus und der Transkription spielen. Eine Beeinträchtigung der Transkriptionsregulation, hauptsächlich durch Chromatinreorganisation, ist ein Mechanismus der zu GB führen kann. Die Ergebnisse dieser Arbeit zeigen, dass hKIAA1202 beim Menschen wahrscheinlich an der Ausbildung kognitiver Fähigkeiten beteiligt ist. Für hFBXO25 dagegen ist dies eher unwahrscheinlich

C57BL/6J mouse hippocampus CA1 pyramidal cell morphologies

<p>title : Reconstruction of hippocampus CA1 cell morphologies</p> <p>specimen : Mus musculus</p> <p>sex : male</p> <p>strain : C57BL/6J</p> <p>age : post-natal day 13-16</p> <p>This neuron was recorded and filled with biocytin (3 mg/ml) in a 300 µm thick coronal slice of rat hippocampus, using 2 - 10 MOhm patch pipettes. 3,3′-diaminobenzidine (DAB) was used for revelation. The slice was fixed and the cell reconstructed with Neurolucida using a 100x oil immersion objective.</p

Inferring and validating mechanistic models of neural microcircuits based on spike-train data

The interpretation of neuronal spike train recordings often relies on abstract statistical models that allow for principled parameter estimation and model selection but provide only limited insights into underlying microcircuits. In contrast, mechanistic models are useful to interpret microcircuit dynamics, but are rarely quantitatively matched to experimental data due to methodological challenges. Here we present analytical methods to efficiently fit spiking circuit models to single-trial spike trains. Using derived likelihood functions, we statistically infer the mean and variance of hidden inputs, neuronal adaptation properties and connectivity for coupled integrate-and-fire neurons. Comprehensive evaluations on synthetic data, validations using ground truth in-vitro and in-vivo recordings, and comparisons with existing techniques demonstrate that parameter estimation is very accurate and efficient, even for highly subsampled networks. Our methods bridge statistical, data-driven and theoretical, model-based neurosciences at the level of spiking circuits, for the purpose of a quantitative, mechanistic interpretation of recorded neuronal population activity

iGIF model with parameters extracted from intracellular recordings.

<p><b>(A)</b> Schematic representation of the iGIF model. The input current is first low-pass filtered by the <i>Passive membrane filter</i> . The resulting signal models the subthreshold membrane potential <i>V</i>(<i>t</i>) and, after subtraction of the firing threshold <i>V</i>_T(<i>t</i>), is transformed into a firing intensity λ(<i>t</i>) by the exponential <i>Escape-rate nonlinearity</i>. Spikes are emitted stochastically and elicit both a <i>Spike-triggered conductance</i> <i>η</i>(<i>t</i>) and a <i>Spike-triggered threshold movement</i> <i>γ</i>(<i>t</i>). In the iGIF model, but not in the GIF model, the firing threshold <i>V</i>_T(<i>t</i>) is coupled to the subthreshold membrane potential (dashed circuit). For that, the membrane potential <i>V</i>(<i>t</i>) is first passed through the nonlinear <i>Threshold coupling</i> function <i>θ</i>_∞(<i>V</i>) and then low-pass filtered by the <i>Threshold filter</i> . <b>(B)-(E)</b> Average parameters extracted from 6 Pyr neurons. Black: iGIF-free, red: iGIF-Na. Gray areas indicate one standard deviation across cells for the iGIF-Na model. <b>(B)</b> Passive membrane filter <i>κ</i>_m(<i>t</i>). Inset: passive membrane timescale <i>τ</i>_m. Open circles: results from individual cells. Bar plot: mean and standard deviation. <b>(C)</b> Spike-triggered conductance <i>η</i>(<i>t</i>). Inset: same data on log-log scales. <b>(D)</b> Spike-triggered threshold movement <i>γ</i>(<i>t</i>). <b>(E)</b> Nonlinear threshold coupling <i>θ</i>_∞(<i>V</i>) (iGIF-free, solid black line; iGIF-Na, solid red line). Spikes are emitted stochastically when the spiking boundary <i>V</i> = <i>V</i>_T is approached. This boundary defines the line where the probability <i>p</i> of emitting a spike during a time bin of Δ<i>T</i> = 0.1 ms reaches <i>p</i> = 0.63. To the right, the probability increases further. In the absence of previous action potentials, the spiking boundary is given by <i>V</i> = <i>θ</i> (dashed black). After an action potential, the spiking boundary instantaneously shifts to the right by <i>γ</i>(0)≈25 mV (dashed gray), and then slowly decays back to <i>V</i> = <i>θ</i>. After each spike, the variables (<i>V</i>(<i>t</i>), <i>θ</i>(<i>t</i>)) are reset to (open circle; error bars denote one standard deviation across cells). The open red circle indicates the Na⁺ half-inactivation voltage <i>V</i>_i, where the threshold becomes sensitive to the membrane potential. Inset: percentage increase in model log-likelihood (<i>LL</i>) of iGIF models with respect to the GIF model. Open circles: model performance on the same cell for iGIF-free and iGIF-Na. Bar plots: mean and standard deviation across neurons. <b>(F)</b> Top: <i>LL</i> percentage increase of the iGIF-free model as a function of <i>τ</i>_<i>θ</i>. Red circle: optimal timescale <i>τ</i>_<i>θ</i>. Data are shown from a typical neuron. Bottom: optimal timescales <i>τ</i>_<i>θ</i> extracted from 6 Pyr neurons. <b>(G)</b> Top: <i>LL</i> percentage increase of the iGIF-Na model as a function of <i>k</i>_<i>i</i> and <i>V</i>_<i>i</i>. The <i>LL</i> increases from dark to light red. Red circle: optimal parameters. Bottom: optimal parameters extracted from 6 different Pyr neurons. Note that the half-inactivation voltages <i>V</i>_<i>i</i> reported in the figure are positively biased due to uncompensated liquid junction potential of 14.5 mV (see <a href="http://www.ploscompbiol.org/article/info:doi/10.1371/journal.pcbi.1004761#sec017" target="_blank">Materials and Methods</a>). The mean and the standard deviational ellipse across cells are shown in red.</p