Aerospace medicine and biology: A continuing bibliography with indexes (supplement 288)

This bibliography lists 190 reports, articles and other documents introduced into the NASA scientific and technical information system in August 1986

De novo development of novel DM1 toxic ncRNA targeting small molecules and its biological evaluation

La distròfia miotònica de tipus 1 (DM1) és un trastorn neuromuscular incurable causat per les transcripcions tòxiques del gen DMPK. Aquests transcrits porten expansions de repeticions CUG a les regions no traduïdes 3′ (3′UTR). La complexitat intrínseca i la falta de dades cristal·logràfiques fan que les regions d'ARN no codificant siguin objectius difícils d'estudiar en el camp del desenvolupament de nous fàrmacs. En el cas de la DM1, els transcrits tòxics tendeixen a estancar-se a l'interior dels nuclis formant complexos cossos d'inclusió anomenats foci i segrestant molts factors de splicing alternatiu essencials com el Muscleblind-like 1 (MBNL1). La majoria de les característiques fenotípiques de la DM1 es deriven de la reduïda disponibilitat de MBNL1 lliure, per la qual cosa molts esforços terapèutics es centren en recuperar la seva activitat regular. Per a això, en la present tesi, decidim utilitzar com a diana terapèutica l'ARN CUG, amb la finalitat d'alliberar MBNL1. Pel que respecta al disseny de noves estructures, es descriu el cribratge in-silico mitjançant tècniques de disseny de fàrmacs basades en estructura usant dues premisses diferents d'abordar CUG. A més, es desenvolupen vies sintètiques per als candidats seleccionats basades en química clic. Finalment, per a avaluar la seva activitat biològica, es posa a punt un assaig bioquímic ja descrit, i s'utilitzen models cel·lulars i cèl·lules musculars derivades de pacients per a avaluar els candidats més prometedors. Els resultats obtinguts poden conduir a posteriors generacions de lligands, posant de manifest un nou tractament assequible contra la DM1.La distrofia miotónica de tipo 1 (DM1) es un trastorno neuromuscular incurable causado por las transcripciones tóxicas del gen DMPK. Estos transcritos llevan expansiones de repeticiones CUG en las regiones no traducidas 3′ (3′UTR). La complejidad intrínseca y la falta de datos cristalográficos hacen que las regiones de ARN no codificante sean objetivos difíciles de estudiar en el campo del desarrollo de nuevos fármacos. En la DM1, los transcritos tóxicos tienden a estancarse en el interior de los núcleos formando complejos cuerpos de inclusión llamados foci y secuestrando muchos factores de splicing alternativo esenciales como el Muscleblind-like 1 (MBNL1). La mayoría de las características fenotípicas de la DM1 se derivan de la reducida disponibilidad de MBNL1 libre, por lo que muchos esfuerzos terapéuticos se centran en recuperar su actividad regular. Para ello, en la presente tesis, decidimos utilizar como diana terapéutica el ARN CUG, con el fin de liberar MBNL1. Por lo que respecta al diseño de nuevas estructuras, se describe el cribado in-silico mediante técnicas de diseño de fármacos basadas en estructura usando dos premisas diferentes de abordar CUG. Además, se desarrollan vías sintéticas para los candidatos seleccionados basadas en química click. Por último, para evaluar su actividad biológica, se pone a punto un ensayo bioquímico ya descrito, y se utilizan modelos celulares y células musculares derivadas de pacientes para evaluar los candidatos más prometedores. Los resultados obtenidos pueden conducir a posteriores generaciones de ligandos, poniendo de manifiesto un nuevo tratamiento asequible contra la DM1.Myotonic Dystrophy type 1 (DM1) is an incurable neuromuscular disorder caused by toxic DMPK transcripts that carry CUG repeat expansions in the 3′ untranslated regions (3′UTR). The intrinsic complexity and lack of crystallographic data make noncoding RNA regions challenging targets to study in the field of drug discovery. In DM1, toxic transcripts tend to stall in the nuclei forming complex inclusion bodies called foci and sequestering many essential alternative splicing factors such as Muscleblind-like 1 (MBNL1). Most DM1 phenotypic features stem from the reduced availability of free MBNL1, and therefore many therapeutic efforts are focused on recovering its regular activity. For that purpose, in the present thesis, we decide to target CUG RNA to free MBNL1. The in-silico screening using structure-based drug design techniques of novel candidates based on two different approaches is described. Furthermore, synthetic pathways are developed for the selected candidates based on the click chemistry approach. Finally, to assess their biological activity, an already described biochemical test is tuned, and cellular models and patient-derived muscular cells are used to evaluate the most promising candidates. The obtained results may lead to subsequent generations of ligands, highlighting a new affordable treatment against DM1

Chromatoid body mediated RNA regulation in mouse male germline

Male germ cell differentiation, spermatogenesis is an exceptional developmental process that produces a massive amount of genetically unique spermatozoa. The complexity of this process along with the technical limitations in the germline research has left many aspects of spermatogenesis poorly understood. Post-meiotic haploid round spermatids possess the most complex transcriptomes of the whole body. Correspondingly, efficient and accurate control mechanisms are necessary to deal with the huge diversity of transcribed RNAs in these cells. The high transcriptional activity in round spermatids is accompanied by the presence of an uncommonly large cytoplasmic ribonucleoprotein granule, called the chromatoid body (CB) that is conjectured to participate in the RNA post-transcriptional regulation. However, very little is known about the possible mechanisms of the CB function. The development of a procedure to isolate CBs from mouse testes was this study’s objective. Anti-MVH immunoprecipitation of cross-linked CBs from a fractionated testicular cell lysate was optimized to yield considerable quantities of pure and intact CBs from mice testes. This protocol produced reliable and reproducible data from the subsequent analysis of CB’s protein and RNA components. We found that the majority of the CB’s proteome consists of RNA-binding proteins that associate functionally with different pathways. We also demonstrated notable localization patterns of one of the CB transient components, SAM68 and showed that its ablation does not change the general composition or structure of the CB. CB-associated RNA analysis revealed a strong accumulation of PIWI-interacting RNAs (piRNAs), mRNAs and long non-coding RNAs (lncRNAs) in the CB. When the CB transcriptome and proteome analysis results were combined, the most pronounced molecular functions in the CB were related to piRNA pathway, RNA post-transcriptional processing and CB structural scaffolding. In addition, we demonstrated that the CB is a target for the main RNA flux from the nucleus throughout all steps of round spermatid development. Moreover, we provided preliminary evidence that those isolated CBs slice target RNAs in vitro in an ATPdependent manner. Altogether, these results make a strong suggestion that the CB functions involve RNA-related and RNA-mediated mechanisms. All the existing data supports the hypothesis that the CB coordinates the highly complex haploid transcriptome during the preparation of the male gametes for fertilization. Thereby, this study provides a fundamental basis for the future functional analyses of ribonucleoprotein granules and offers also important insights into the mechanisms governing male fertility.Miesten sukusolujen kehittyminen, spermatogeneesi, on tärkeä ja monella tavoin poikkeuksellinen kehitysprosessi, joka tuottaa suunnattomia määriä geneettisesti yksilöllisiä siittiöitä. Spermatogeneesi on monimutkainen, tarkasti säädelty tapahtumasarja, mikä myös aiheuttaa teknisiä haasteita spermatogeneesin molekyylimekanismien tutkimisessa. Näin ollen prosessin yksityiskohdat ovat vielä pitkälti tuntemattomia. Eräs siittiön haploidien esiasteiden (pyöreät spermatidit) erityisominaisuus on niiden ainutlaatuisen runsas transkriptionaalinen aktiivisuus. Genomin aktiivinen ilmentyminen puolestaan edellyttää tehokkaita ja täsmällisiä RNA:n säätelymekanismeja. Pyöreiden spermatidien solulimassa sijaitsee epätavallisen suuri RNA:ta ja proteiineja sisältävä rakenne, kromatoidikappale (chromatoid body, CB), joka ilmaantuu juuri voimakkaimman transkriptioaallon aikana ja osallistuu RNA-säätelyyn. Tutkimuksen tavoitteena oli selvittää CB:n toimintaa siittiönkehityksen aikana. Tärkeänä osana tutkimusta kehitimme menetelmän, jonka avulla CB:t voidaan eristää hiiren kiveksestä. Menetelmä on yksinkertainen, nopea ja tehokas, ja sen avulla saadaan eristettyä toistettavasti rakenteeltaan ehjiä CB:ta, joiden puhtaus on hyvä ja määrä riittävä molekyylitason analyysiin. Jatkotutkimukset paljastivat, että suurin osa CB:n sisältämistä proteiineista on erilaisilla RNA-säätelyreiteillä toimivia RNA:ta sitovia proteiineja. Useat CB:n proteiineista pysyvät rakenteessa stabiilisti, mutta näytimme myös, että RNA:ta sitova proteiini SAM68 vierailee CB:ssa vain hyvin hetkellisesti tarkasti määritellyssä kehitysvaiheessa. Poistogeenisen hiirimallin avulla saimme selville, ettei SAM68 proteiinia kuitenkaan tarvita CB:n muodostumiseen. Tulostemme mukaan CB:een kulkeutuu suuri määrä RNA:ta kaikissa pyöreiden spermatidien kehitysvaiheissa. RNA-sekvenointi osoitti, että pienet piRNA (PIWI-interacting RNA) molekyylit ovat rikastuneet CB:ssa. Lisäksi CB sisältää suuren joukon erilaisia lähetti-RNA:ita ja aivan tuntemattomia intergeenisiä eikoodaavia RNA:ita. Analyysimme mukaan hallitsevin CB:n molekyylireiteistä on piRNAvälitteinen RNA-säätelyreitti, mutta myös lähetti-RNA:n prosessointiin liittyvät tekijät ovat vahvasti edustettuina. CB ei selvästikään ole vain passiivinen RNA:n varastointipaikka, vaan näytimme sen kykenevän ATP:stä riippuvaiseen RNA prosessointiin in vitro Kaikki tutkimuksen tulokset osoittavat, että CB on keskeinen RNA:n säätelykeskus, joka koordinoi sukusolujen erittäin monimuotoista transkriptomia. Tällä toiminnallaan CB osallistuu tärkeänä tekijänä miesten hedelmällisyyden ja sukusolujen geneettisen ja epigeneettisen informaation säätelyyn.Siirretty Doriast

Aerospace medicine and biology: A continuing bibliography with indexes (supplement 297)

This bibliography lists 89 reports, articles and other documents introduced into the NASA scientific and technical information system in April, 1987

The Dollar General: Continuous Custom Gesture Recognition Techniques At Everyday Low Prices

Humans use gestures to emphasize ideas and disseminate information. Their importance is apparent in how we continuously augment social interactions with motion—gesticulating in harmony with nearly every utterance to ensure observers understand that which we wish to communicate, and their relevance has not escaped the HCI community\u27s attention. For almost as long as computers have been able to sample human motion at the user interface boundary, software systems have been made to understand gestures as command metaphors. Customization, in particular, has great potential to improve user experience, whereby users map specific gestures to specific software functions. However, custom gesture recognition remains a challenging problem, especially when training data is limited, input is continuous, and designers who wish to use customization in their software are limited by mathematical attainment, machine learning experience, domain knowledge, or a combination thereof. Data collection, filtering, segmentation, pattern matching, synthesis, and rejection analysis are all non-trivial problems a gesture recognition system must solve. To address these issues, we introduce The Dollar General (TDG), a complete pipeline composed of several novel continuous custom gesture recognition techniques. Specifically, TDG comprises an automatic low-pass filter tuner that we use to improve signal quality, a segmenter for identifying gesture candidates in a continuous input stream, a classifier for discriminating gesture candidates from non-gesture motions, and a synthetic data generation module we use to train the classifier. Our system achieves high recognition accuracy with as little as one or two training samples per gesture class, is largely input device agnostic, and does not require advanced mathematical knowledge to understand and implement. In this dissertation, we motivate the importance of gestures and customization, describe each pipeline component in detail, and introduce strategies for data collection and prototype selection

Advances in Human-Robot Interaction

Rapid advances in the field of robotics have made it possible to use robots not just in industrial automation but also in entertainment, rehabilitation, and home service. Since robots will likely affect many aspects of human existence, fundamental questions of human-robot interaction must be formulated and, if at all possible, resolved. Some of these questions are addressed in this collection of papers by leading HRI researchers

Aerospace medicine and biology: A continuing bibliography with indexes (supplement 286)

This bibliography lists 213 reports, articles and other documents introduced into the NASA scientific and technical information system in June 1986

Proceedings of the Seventeenth Annual Conference on Manual Control

Manual control is considered, with concentration on perceptive/cognitive man-machine interaction and interface

Advanced Knowledge Application in Practice

The integration and interdependency of the world economy leads towards the creation of a global market that offers more opportunities, but is also more complex and competitive than ever before. Therefore widespread research activity is necessary if one is to remain successful on the market. This book is the result of research and development activities from a number of researchers worldwide, covering concrete fields of research