drosophila-preparation-longitudinal-imaging-heart-function-vivo-using

Drosophila Preparation and Longitudinal Imaging of Heart Function In Vivo Using Optical Coherence Microscopy (OCM)

Longitudinal study of the heartbeat in small animals contributes to understanding structural and functional changes during heart development. Optical ...

quantitative-analysis-neuronal-dendritic-arborization-complexity

Quantitative Analysis of Neuronal Dendritic Arborization Complexity in Drosophila

Dendrites are the branched projections of a neuron, and dendritic morphology reflects synaptic organization during the development of the nervous system. ...

Massachusetts General Hospital and Harvard Medical School

Mass General Institute of Neurodegenerative Disease

Most reports on the natural history, manifestations, and treatment of polycystic liver disease are based on the disease as it manifests in patients with autosomal dominant polycystic kidney disease (ADPKD). The purpose of this study was to develop a clinical profile of isolated autosomal dominant polycystic liver disease (ADPLD) using nonaffected family members as controls. The study included 146 probands, known affected relatives, and first-degree relatives of affected individuals. Participants underwent a formalized medical history interview and physical examination, ultrasonographic examination of the liver and kidneys, magnetic resonance angiography of the brain, and echocardiography. Thirty-eight of the 49 individuals diagnosed with polycystic liver disease before participation in the study were or had been symptomatic. Of 97 previously undiagnosed at-risk individuals, 23 were affected, 39 were unaffected, and 35 were indeterminate. Compared with patients with a negative or indeterminate diagnosis, those with polycystic liver disease had slightly higher levels of serum alkaline phosphatase and total bilirubin and lower levels of total cholesterol and triglycerides. Female patients had a significantly higher mean cyst score than male patients. The cysts were found to arise from the dilatation of biliary microhamartomas and from peribiliary glands. Structural mitral leaflet abnormalities were detected more frequently in affected than in indeterminate or nonaffected individuals. A vascular phenotype was detected in 5.6% of the patients with isolated ADPLD diagnosed clinically and/or by linkage analysis but in none of the unaffected patients. In conclusion, isolated ADPLD is underdiagnosed and genetically distinct from polycystic liver disease associated with ADPKD but with similar pathogenesis, manifestations, and management.

Clinical profile of autosomal dominant polycystic liver disease.

Autosomal dominant polycystic liver disease (ADPLD) is a distinct clinical and genetic entity that can occur independently from autosomal dominant polycystic kidney disease (ADPKD). We previously studied two large kindreds and reported localization of a gene for ADPLD to an approximately 8-Mb region, flanked by markers D19S586/D19S583 and D19S593/D19S579, on chromosome 19p13.2-13.1. Expansion of these kindreds and identification of an additional family allowed us to define flanking markers CA267 and CA048 in an approximately 3-Mb region containing >70 candidate genes. We used a combination of denaturing high-performance liquid chromatography (DHPLC) heteroduplex analysis and direct sequencing to screen a panel of 15 unrelated affected individuals for mutations in genes from this interval. We found sequence variations in a known gene, PRKCSH, that were not observed in control individuals, that segregated with the disease haplotype, and that were predicted to be chain-terminating mutations. In contrast to PKD1, PKD2, and PKHD1, PRKCSH encodes a previously described human protein termed "protein kinase C substrate 80K-H" or "noncatalytic beta-subunit of glucosidase II." This protein is highly conserved, is expressed in all tissues tested, and contains a leader sequence, an LDLa domain, two EF-hand domains, and a conserved C-terminal HDEL sequence. Its function may be dependent on calcium binding, and its putative actions include the regulation of N-glycosylation of proteins and signal transduction via fibroblast growth-factor receptor. In light of the focal nature of liver cysts in ADPLD, the apparent loss-of-function mutations in PRKCSH, and the two-hit mechanism operational in dominant polycystic kidney disease, ADPLD may also occur by a two-hit mechanism.

Mutations in PRKCSH cause isolated autosomal dominant polycystic liver disease.

Mutations to the prototypical members of the two general classes of polycystins, polycystin-1 encoded by PKD1 and polycystin-2 encoded by PKD2, underlie autosomal-dominant polycystic kidney disease. Here we report the identification of a pair of genes homologous to PKD1 from both the human and mouse genomes. PKD1L2 and PKD1L3 are located on human chromosome 16q22-q23 and mouse chromosome 8 and are alternatively spliced. The human and mouse forms of PKD1L2 are highly conserved, with each one consisting of 43 exons and approximately 2,460 codons. PKD1L3 shows regional sequence divergence, with the mouse form having two additional exons and a much larger exon 5. The predicted protein products of PKD1L2 and PKD1L3 contain the combination of GPS and PLAT/LH2 domains that uniquely define them as polycystin-1 family members. They are predicted to have 11 membrane-spanning regions with a large extracellular domain consistent with the proposed receptor function of this protein family. PKD1L2 and PKD1L3 contain strong ion channel signature motifs that suggest their possible function as components of cation channel pores. Polycystin-1-related proteins may not only regulate channels, but may actually be part of the pore-forming unit.

Identification of two novel polycystic kidney disease-1-like genes in human and mouse genomes.

Mutations in PRKCSH, encoding the beta-subunit of glucosidase II, an N-linked glycan-processing enzyme in the endoplasmic reticulum (ER), cause autosomal dominant polycystic liver disease. We found that mutations in SEC63, encoding a component of the protein translocation machinery in the ER, also cause this disease. These findings are suggestive of a role for cotranslational protein-processing pathways in maintaining epithelial luminal structure and implicate noncilial ER proteins in human polycystic disease.

Mutations in SEC63 cause autosomal dominant polycystic liver disease.

UBQLN1 variants have been associated with increased risk for late-onset Alzheimer's disease (AD). We produced transgenic Drosophila models that either silence (by RNAi) or overexpress the Drosophila ortholog of human UBQLN1, dUbqln. Silencing of dUbqln in the central nervous system led to age-dependent neurodegeneration and shortened lifespan. Silencing of dUbqln in the wing led to wing vein loss that could be partially rescued by mutant rhomboid (rho), a known component of epidermal growth factor receptor signaling pathway. Conversely, overexpression of dUbqln promoted ecotopic wing veins. Overexpression of dUbqln in the eye rescued a small, rough eye phenotype induced by overexpression of Drosophila presenilin (dPsn), and also rescuing dPsn-induced malformations in bristles. In contrast, RNAi silencing of dUbqln enhanced the retinal degenerative defect induced by overexpression of dPsn. Finally, co-overexpression of dUbqln and the human amyloid precursor protein (APP) in the eye significantly reduced the levels of full-length APP and its C-terminal fragment. Collectively, these data support in vivo functional interaction between UBQLN1 and the AD-associated genes, presenilin and APP, and provide further clues regarding the potential role of UBQLN1 in AD pathogenesis.

Isolation and characterization of the Drosophila ubiquilin ortholog dUbqln: in vivo interaction with early-onset Alzheimer disease genes.

&#x8109;&#x51B2;&#x7684;&#x573A;&#x51DD;&#x80F6;&#x7535;&#x6CF3; (&#x8109;&#x51B2;&#x573A;&#x51DD;&#x80F6;&#x7535;&#x6CF3;) &#x65B9;&#x6CD5;&#x5DF2;&#x6210;&#x4E3A; DNA &#x5E7F;&#x6CDB;&#x8303;&#x56F4;&#x7684;&#x5206;&#x6790;&#x5E94;&#x7528;&#x7A0B;&#x5E8F;&#x4E2D;&#x7684;&#x6807;&#x51C6;&#x5DE5;&#x5177;&#x3002;&#x4F46;&#x8109;&#x51B2;&#x7684;&#x573A;&#x6761;&#x4EF6;&#x4E0B; DNA &#x8FC1;&#x79FB;&#x73B0;&#x8C61;&#x7684;&#x8BB8;&#x591A;&#x65B9;&#x9762;&#x5728;&#x7535;&#x573A;&#x4F5C;&#x7528;&#x4FDD;&#x6301;&#x4E0D;&#x53D8;&#x7684;&#x66F4;&#x591A;&#x5E38;&#x89C4;&#x60C5;&#x51B5;&#x6BD4;&#x8F83;&#x4E0D;&#x597D;&#x7406;&#x89E3;&#x3002;&#x5BF9;&#x4E8E;&#x8FD9;&#x79CD;&#x7F3A;&#x9677;&#x7684;&#x4E3B;&#x8981;&#x539F;&#x56E0;&#x662F;&#x8109;&#x51B2;&#x573A;&#x51DD;&#x80F6;&#x7535;&#x6CF3;&#x5B9E;&#x9A8C;&#x662F;&#x7E41;&#x7410;&#x6781;&#x957F;&#x65F6;&#x95F4; (&#x7EA6; 10-15 h) &#x5206;&#x79BB;&#x548C;&#x9700;&#x8981;&#x901A;&#x8FC7; poststaining &#x7684;&#x8FD0;&#x884C;&#x5B8C;&#x6210;&#x540E;&#x6267;&#x884C;&#x51DD;&#x80F6;&#x5206;&#x6790;&#x6267;&#x884C;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x4ECB;&#x7ECD;&#x6613;&#x4E8E;&#x751F;&#x6210;&#x5FAE;&#x578B;&#x7684;&#x677F;&#x51DD;&#x80F6;&#x5668;&#x5177;&#xFF0C;&#x4EE5;&#x5904;&#x7406;&#x8FD9;&#x4E9B;&#x95EE;&#x9898;&#xFF0C;&#x901A;&#x8FC7;&#x542F;&#x7528;&#x5927; DNA &#x7247;&#x6BB5;&#x7684;&#x957F;&#x5EA6;&#x5FC5;&#x987B;&#x5206;&#x5F00;&#x573A;&#x53CD;&#x6F14;&#x51DD;&#x80F6;&#x7535;&#x6CF3; (FIGE) &#x5728; 60-90 &#x5206;&#x949F;&#x8FBE; 35 kb&#x3002;&#x5C3A;&#x5BF8;&#x7D27;&#x51D1;&#x7684;&#x8BBE;&#x5907;&#x8FD8;&#x5141;&#x8BB8;&#x6574;&#x4E2A;&#x51DD;&#x80F6;&#xFF0C;&#x4E0D;&#x65AD;&#x8FDB;&#x884C;&#x76D1;&#x6D4B;&#xFF0C;&#x4EE5;&#x4FBF;&#x53EF;&#x4EE5;&#x4F7F;&#x7528;&#x9AD8;&#x5206;&#x8FA8;&#x7387;&#x7684; CCD &#x76F8;&#x673A;&#x5B9E;&#x65F6;&#x6210;&#x50CF;&#x5206;&#x79BB;&#x8FC7;&#x7A0B;&#x3002;&#x5916;&#x52A0;&#x7684;&#x7535;&#x538B;&#x6CE2;&#x5F62;&#x4EFB;&#x610F;&#x63A7;&#x5236;&#x662F;&#x4F7F;&#x7528;&#x4E0E;&#x9AD8;&#x7535;&#x538B;&#x653E;&#x5927;&#x5668;&#x63A5;&#x53E3;&#x51FD;&#x6570;&#x751F;&#x6210;&#x5668;&#x5B9E;&#x73B0;&#x7684;&#x3002;&#x8FD9;&#x4E9B;&#x529F;&#x80FD;&#x4F7F;&#x6211;&#x4EEC;&#x80FD;&#x591F;&#x63A2;&#x7D22;&#x4E0E; DNA &#x8FC1;&#x79FB; &#xFF08;&#x6D41;&#x52A8;&#xFF0C;&#x6269;&#x6563;&#x548C;&#x5206;&#x79BB;&#x51B3;&#x8BAE;&#xFF09; &#x76F8;&#x5173;&#x8054;&#x7684;&#x57FA;&#x672C;&#x7269;&#x7406;&#x53C2;&#x6570;&#x7684;&#x5927;&#x5C0F;&#x4F9D;&#x8D56;&#x3002;&#x8FD9;&#x4E9B;&#x6570;&#x636E;&#x663E;&#x793A;&#x5728;&#x5206;&#x79BB;&#x7684;&#x5206;&#x8FA8;&#x7387;&#x589E;&#x52A0;&#x7531;&#x4E8E;&#x6D41;&#x52A8;&#x6027;&#x548C;&#x6269;&#x6563;&#x7684;&#x6709;&#x5229;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x7684; DNA &#x7247;&#x6BB5;&#x5927;&#x5C0F;&#x4E0E;&#x73B0;&#x884C;&#x5947;&#x602A;&#x5236;&#x5EA6;&#xFF0C;&#x5E76;&#x7A81;&#x51FA;&#x663E;&#x793A; &#xFF08;&#x5F88;&#x5C11;&#x91CF;&#x5316;&#x53C2;&#x6570;&#xFF09; &#x6269;&#x6563;&#x65B9;&#x9762;&#x7684;&#x91CD;&#x8981;&#x4F5C;&#x7528;&#x3002;&#x9664;&#x4E86;&#x6BD4;&#x5E38;&#x89C4;&#x4EEA;&#x5668;&#x5FEB;&#x662F;&#x4E00;&#x4E2A;&#x6570;&#x91CF;&#x7EA7;&#x7684;&#x5206;&#x79BB;&#x65F6;&#x95F4;&#x5E26;&#x6765;&#x5B9E;&#x9645;&#x7684;&#x5229;&#x76CA;&#xFF0C;&#x8FD9;&#x4E9B;&#x7814;&#x7A76;&#x7684;&#x7ED3;&#x679C;&#x63D0;&#x4F9B;&#x4EE5;&#x524D;&#x4E0D;&#x53EF;&#x7528;&#x7684;&#x7406;&#x6027;&#x57FA;&#x7840;&#xFF0C;&#x786E;&#x5B9A;&#x6700;&#x4F18;&#x7684;&#x5206;&#x79BB;&#x6761;&#x4EF6;&#xFF0C;&#x4ECE;&#x800C;&#x4FC3;&#x8FDB;&#x7406;&#x89E3;&#x57FA;&#x672C;&#x7269;&#x7406;&#x8FC7;&#x7A0B;&#x63A7;&#x5236; DNA &#x7535;&#x6CF3;&#x8109;&#x51B2;&#x9886;&#x57DF;&#x4E2D;&#x7684;&#x65B0;&#x89C1;&#x89E3;&#x3002;

&#x5FAE;&#x578B;&#x7684;&#x7CFB;&#x7EDF;&#x5FEB;&#x901F;&#x573A;&#x53CD;&#x6F14;&#x7684;&#x51DD;&#x80F6;&#x7535;&#x6CF3;&#x7684; DNA &#x4E0E;&#x6574;&#x4E2A;&#x51DD;&#x80F6;&#x7684;&#x5B9E;&#x65F6;&#x68C0;&#x6D4B;&#x3002;

Pulsed field gel electrophoresis (PFGE) methods have become standard tools in a wide range of DNA analysis applications. But many aspects of DNA migration phenomena under pulsed field conditions are not well understood as compared with the more conventional situation where the electric field is held constant. A key reason for this deficiency is that PFGE experiments are cumbersome to perform due to extremely long separation times (approximately 10-15 h) and the need to perform gel analysis by poststaining after completion of the run. Here we introduce an easy to build miniaturized slab gel apparatus that addresses these issues by enabling large DNA fragments up to 35 kb in length to be separated using field inversion gel electrophoresis (FIGE) in 60-90 min. The compact size of the device also allows the entire gel to be continuously monitored so that the separation processes can be imaged in real time using a high-resolution CCD camera. Arbitrary control over the applied voltage waveforms is achieved using a function generator interfaced with a high voltage amplifier. These capabilities allow us to probe the size dependence of fundamental physical parameters associated with DNA migration (mobility, diffusion, and separation resolution). These data reveal a surprising regime where separation resolution increases with DNA fragment size owing to a favorable interplay between mobility and diffusion scalings and highlight the important role of diffusion (a seldom quantified parameter). In addition to the practical benefit of separation times that are an order of magnitude faster than conventional instruments, the results of these studies provide a previously unavailable rational basis to identify optimal separation conditions and contribute new insights toward understanding the underlying physical processes that govern DNA electrophoresis in pulsed fields.

Miniaturized system for rapid field inversion gel electrophoresis of DNA with real-time whole-gel detection.

Champ puls&#xE9; &#xE9;lectrophor&#xE8;se sur gel (PFGE) m&#xE9;thodes sont devenus des outils standards dans un large &#xE9;ventail d&#x26;#39;applications d&#x26;#39;analyse d&#x26;#39;ADN. Mais de nombreux aspects de ph&#xE9;nom&#xE8;nes de migration d&#x26;#39;ADN dans des conditions de champ puls&#xE9;s ne sont pas bien compris par rapport &#xE0; la situation plus classique o&#xF9; le champ &#xE9;lectrique est maintenue constante. Une des principales raisons de cette carence est que les exp&#xE9;riences en PFGE sont difficiles &#xE0; r&#xE9;aliser en raison de temps de s&#xE9;paration extr&#xEA;mement longue (environ 10-15 h) et la n&#xE9;cessit&#xE9; d&#x26;#39;effectuer une analyse de gel par poststaining apr&#xE8;s la fin de la course. Ici, nous introduisons un outil facile &#xE0; construire miniaturis&#xE9; appareils plaque de gel qui aborde ces questions en permettant grands fragments d&#x26;#39;ADN jusqu&#x26;#39;&#xE0; 35 kb de longueur pour &#xEA;tre s&#xE9;par&#xE9;s par &#xE9;lectrophor&#xE8;se sur gel en champ d&#x26;#39;inversion (FIGE) en 60-90 min. La taille compacte de l&#x26;#39;appareil permet &#xE9;galement le gel entier &#xE0; &#xEA;tre surveill&#xE9;s en permanence afin que les proc&#xE9;d&#xE9;s de s&#xE9;paration peuvent &#xEA;tre visualis&#xE9;s en temps r&#xE9;el en utilisant une cam&#xE9;ra CCD haute r&#xE9;solution. Contr&#xF4;le arbitraire sur les formes d&#x26;#39;onde de tension appliqu&#xE9;e est r&#xE9;alis&#xE9;e en utilisant un g&#xE9;n&#xE9;rateur de fonction interfac&#xE9; avec un amplificateur de haute tension. Ces capacit&#xE9;s nous permettent de sonder la d&#xE9;pendance &#xE0; la taille des param&#xE8;tres fondamentaux physiques associ&#xE9;es &#xE0; l&#x26;#39;ADN des migrations (mobilit&#xE9;, la diffusion et la r&#xE9;solution de la s&#xE9;paration). Ces donn&#xE9;es r&#xE9;v&#xE8;lent un r&#xE9;gime &#xE9;tonnant o&#xF9; la r&#xE9;solution augmente de s&#xE9;paration avec raison la taille du fragment d&#x26;#39;ADN &#xE0; une interaction favorable entre la mobilit&#xE9; et de diffusion battitures et mettre en &#xE9;vidence le r&#xF4;le important de la diffusion (un param&#xE8;tre rarement quantifi&#xE9;s). En plus de l&#x26;#39;avantage pratique de temps de s&#xE9;paration qui sont d&#x26;#39;un ordre de grandeur plus rapide que les instruments classiques, les r&#xE9;sultats de ces &#xE9;tudes fournissent une base rationnelle non disponibles auparavant pour identifier les conditions optimales de s&#xE9;paration et de contribuer de nouvelles perspectives &#xE0; la compr&#xE9;hension des processus physiques sous-jacents qui r&#xE9;gissent &#xE9;lectrophor&#xE8;se de l&#x26;#39;ADN dans les champs puls&#xE9;s.

Syst&#xE8;me miniaturis&#xE9; pour &#xE9;lectrophor&#xE8;se en champ Gel Rapid Inversion de l&#x26;#39;ADN avec en temps r&#xE9;el entier gel de d&#xE9;tection

Gepulste Feld-Gel-Elektrophorese (PFGE)-Methoden sind standard-Tools in eine breite Palette von DNA-Analyse-Anwendungen geworden. Aber viele Aspekte der DNA-Migration-Ph&auml;nomene unter Feldbedingungen gepulste sind nicht gut verstanden, im Vergleich zu der konventionellen Situation, wo das elektrische Feld konstant gehalten wird. Ein wesentlicher Grund f&uuml;r diesen Mangel ist, dass PFGE Experimente umst&auml;ndlich durch extrem lange Trennung Zeiten (ca. 10-15 h) und die Notwendigkeit f&uuml;r Gel-Analyse von poststaining nach Abschluss der Ausf&uuml;hrung durchf&uuml;hren. Hier stellen wir eine leicht zu bauen miniaturisierte Tafel-Gel, den Apparat, der diese Fragen behandelt, indem es erm&ouml;glicht gro&szlig;e DNA-Fragmente bis zu 35 kb L&auml;nge mit Feld-Umkehrung-Gel-Elektrophorese (Feigenbawm) in 60-90 min getrennt werden. Die kompakte Gr&ouml;&szlig;e des Ger&auml;ts kann auch das gesamte Gel kontinuierlich &uuml;berwacht werden, so dass die Trennung Prozesse in Echtzeit mit eine hochaufl&ouml;sende CCD-Kamera abgebildet werden k&ouml;nnen. Willk&uuml;rliche Kontrolle &uuml;ber die angewandte Spannung-Wellenformen geschieht &uuml;ber einen Funktionsgenerator mit einem Hochspannungs-Verst&auml;rker angeschlossen. Diese Funktionen erlauben uns, die Abh&auml;ngigkeit der Gr&ouml;&szlig;e von grundlegenden physikalischen Parametern zugeordnet DNA-Migration (Mobilit&auml;t, Verbreitung und Trennung Aufl&ouml;sung) Sonde. Diese Daten zeigen eine &uuml;berraschende Regelung in der Trennung Aufl&ouml;sung mit DNA Fragmentgr&ouml;&szlig;e aufgrund einer g&uuml;nstigen Wechselspiel zwischen Mobilit&auml;t und Verbreitung Zunder steigt und die wichtige Rolle der Diffusion (selten quantifizierte Parameter) zu markieren. Neben den praktischen Nutzen der Trennung-Zeiten, die eine Gr&ouml;&szlig;enordnung schneller als konventionelle Instrumente sind, bieten die Ergebnisse dieser Studien eine zuvor nicht rationale Grundlage, um optimale Trennung Bedingungen zu identifizieren und neue Erkenntnisse zum Verst&auml;ndnis der zugrunde liegenden physikalischen Prozesse &uuml;ber den DNA-Elektrophorese in gepulster Felder beitragen.

Miniaturisierte System f&uuml;r schnelle Feld Umkehrung gel Elektrophorese von DNA mit ganz-Gel-Erkennung in Echtzeit.

Pulsed field gel electrophoresis (attrezzature) metodi sono diventati strumenti standard nelle applicazioni di analisi di una vasta gamma di DNA. Ma molti aspetti dei fenomeni di migrazione del DNA in condizioni di campo pulsato non sono ben chiari rispetto alla situazione pi&ugrave; convenzionale, dove il campo elettrico &egrave; mantenuto costante. Un motivo chiave per questa carenza &egrave; che attrezzature esperimenti sono ingombranti per eseguire a causa della separazione estremamente lunghi periodi (circa 10-15 h) e la necessit&agrave; di eseguire analisi del gel di poststaining dopo il completamento della corsa. Qui presentiamo un facile costruire gel lastra miniaturizzato apparato che risolve questi problemi attivando grande DNA frammenti fino a 35 kb in lunghezza per essere separati mediante elettroforesi del gel di inversione di campo (FIGE) in 60-90 min. Le dimensioni compatte del dispositivo permettono anche il gel intero essere continuamente monitorato affinch&eacute; i processi di separazione possono essere imaged in tempo reale utilizzando una telecamera CCD ad alta risoluzione. Arbitrario controllo sopra le forme d&#39;onda di tensione applicata &egrave; realizzato utilizzando un generatore di funzione interfacciato con un amplificatore ad alta tensione. Queste capacit&agrave; ci permettono di sondare la dipendenza della dimensione dei fondamentali parametri fisici associati con la migrazione del DNA (mobilit&agrave;, diffusione e separazione risoluzione). Questi dati rivelano un sorprendente regime dove risoluzione separazione aumenta con la dimensione del frammento di DNA a causa di un&#39;interazione favorevole tra mobilit&agrave; e diffusione scaglie ed evidenziano l&#39;importante ruolo di diffusione (un parametro raramente quantificato). Oltre al vantaggio pratico dei tempi di separazione che sono un ordine di grandezza superiore rispetto agli strumenti convenzionali, i risultati di questi studi forniscono una base razionale precedentemente non disponibile per identificare le condizioni ottimali di separazione e contribuire con nuove intuizioni verso la comprensione dei processi fisici sottostanti che governano l&#39;elettroforesi del DNA in campi pulsati.

Sistema miniaturizzato per inversione di campo rapid gel elettroforesi di DNA con rilevamento in tempo reale di tutto-gel.

&#x30D1;&#x30EB;&#x30B9;&#x30D5;&#x30A3;&#x30FC;&#x30EB;&#x30C9;&#x30B2;&#x30EB;&#x96FB;&#x6C17;&#x6CF3;&#x52D5;&#xFF08;PFGE&#xFF09;&#x30E1;&#x30BD;&#x30C3;&#x30C9;&#x306F;&#x3001;DNA&#x5206;&#x6790;&#x30A2;&#x30D7;&#x30EA;&#x30B1;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x306E;&#x5E83;&#x3044;&#x7BC4;&#x56F2;&#x3067;&#x6A19;&#x6E96;&#x7684;&#x306A;&#x30C4;&#x30FC;&#x30EB;&#x3068;&#x306A;&#x3063;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x96FB;&#x754C;&#x304C;&#x4E00;&#x5B9A;&#x306B;&#x4FDD;&#x305F;&#x308C;&#x3066;&#x3044;&#x308B;&#x5F93;&#x6765;&#x306E;&#x72B6;&#x6CC1;&#x3068;&#x6BD4;&#x8F03;&#x3057;&#x3066;&#x3001;&#x3057;&#x304B;&#x3057;&#x3001;&#x30D1;&#x30EB;&#x30B9;&#x30D5;&#x30A3;&#x30FC;&#x30EB;&#x30C9;&#x6761;&#x4EF6;&#x4E0B;&#x3067;&#x306E;DNA&#x306E;&#x79FB;&#x884C;&#x73FE;&#x8C61;&#x306E;&#x591A;&#x304F;&#x306E;&#x5074;&#x9762;&#x306F;&#x3088;&#x304F;&#x7406;&#x89E3;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x305B;&#x3093;&#x3002;&#x3053;&#x306E;&#x6B20;&#x4E4F;&#x306E;&#x4E3B;&#x306A;&#x7406;&#x7531;&#x306F;&#x3001;PFGE&#x5B9F;&#x9A13;&#x306F;&#x975E;&#x5E38;&#x306B;&#x9577;&#x3044;&#x5206;&#x96E2;&#x6642;&#x9593;&#xFF08;&#x7D04;10&#x301C;15 h&#xFF09;&#x3068;&#x3001;&#x5B9F;&#x884C;&#x306E;&#x5B8C;&#x4E86;&#x5F8C;&#x306B;poststaining&#x306B;&#x3088;&#x3063;&#x3066;&#x30B2;&#x30EB;&#x5206;&#x6790;&#x3092;&#x5B9F;&#x884C;&#x3059;&#x308B;&#x5FC5;&#x8981;&#x304C;&#x3042;&#x308B;&#x305F;&#x3081;&#x3001;&#x5B9F;&#x884C;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x9762;&#x5012;&#x3067;&#x3042;&#x308B;&#x3068;&#x3044;&#x3046;&#x3053;&#x3068;&#x3067;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;60&#x301C;90&#x5206;&#x3067;&#x30D5;&#x30A3;&#x30FC;&#x30EB;&#x30C9;&#x53CD;&#x8EE2;&#x30B2;&#x30EB;&#x96FB;&#x6C17;&#x6CF3;&#x52D5;&#xFF08;FIGE&#xFF09;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x5206;&#x96E2;&#x3059;&#x308B;&#x9577;&#x3055;&#x306F;35&#x30AD;&#x30ED;&#x30D0;&#x30A4;&#x30C8;&#x306B;&#x5927;&#x304D;&#x306A;DNA&#x65AD;&#x7247;&#x3092;&#x6709;&#x52B9;&#x306B;&#x3059;&#x308B;&#x3053;&#x3068;&#x306B;&#x3088;&#x3063;&#x3066;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x554F;&#x984C;&#x306B;&#x5BFE;&#x51E6;&#x5C0F;&#x578B;&#x30B9;&#x30E9;&#x30D6;&#x30B2;&#x30EB;&#x88C5;&#x7F6E;&#x3092;&#x69CB;&#x7BC9;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x7C21;&#x5358;&#x306B;&#x7D39;&#x4ECB;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x88C5;&#x7F6E;&#x306E;&#x30B3;&#x30F3;&#x30D1;&#x30AF;&#x30C8;&#x30B5;&#x30A4;&#x30BA;&#x306F;&#x200B;&#x200B;&#x3001;&#x30B2;&#x30EB;&#x5168;&#x4F53;&#x304C;&#x9023;&#x7D9A;&#x7684;&#x306B;&#x5206;&#x96E2;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x306F;&#x3001;&#x9AD8;&#x89E3;&#x50CF;&#x5EA6;CCD&#x30AB;&#x30E1;&#x30E9;&#x3092;&#x7528;&#x3044;&#x3066;&#x30EA;&#x30A2;&#x30EB;&#x30BF;&#x30A4;&#x30E0;&#x3067;&#x753B;&#x50CF;&#x5316;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x308B;&#x3088;&#x3046;&#x306B;&#x76E3;&#x8996;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x5370;&#x52A0;&#x96FB;&#x5727;&#x6CE2;&#x5F62;&#x4E0A;&#x306E;&#x4EFB;&#x610F;&#x306E;&#x30B3;&#x30F3;&#x30C8;&#x30ED;&#x30FC;&#x30EB;&#x306F;&#x3001;&#x9AD8;&#x96FB;&#x5727;&#x30A2;&#x30F3;&#x30D7;&#x3068;&#x306E;&#x30A4;&#x30F3;&#x30BF;&#x30D5;&#x30A7;&#x30FC;&#x30B9;&#x30D5;&#x30A1;&#x30F3;&#x30AF;&#x30B7;&#x30E7;&#x30F3;&#xB7;&#x30B8;&#x30A7;&#x30CD;&#x30EC;&#x30FC;&#x30BF;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x5B9F;&#x73FE;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x6A5F;&#x80FD;&#x306F;&#x3001;&#x79C1;&#x305F;&#x3061;&#x306F;&#x3001;DNA&#x306E;&#x79FB;&#x884C;&#xFF08;&#x79FB;&#x52D5;&#x3001;&#x62E1;&#x6563;&#x3001;&#x5206;&#x96E2;&#x5206;&#x89E3;&#x80FD;&#xFF09;&#x306B;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x57FA;&#x672C;&#x7684;&#x306A;&#x7269;&#x7406;&#x30D1;&#x30E9;&#x30E1;&#x30FC;&#x30BF;&#x306E;&#x30B5;&#x30A4;&#x30BA;&#x4F9D;&#x5B58;&#x6027;&#x3092;&#x8ABF;&#x3079;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x30C7;&#x30FC;&#x30BF;&#x306F;&#x3001;&#x5206;&#x96E2;&#x5206;&#x89E3;&#x80FD;&#x306F;&#x30E2;&#x30D3;&#x30EA;&#x30C6;&#x30A3;&#x3068;&#x62E1;&#x6563;&#x306E;&#x30B9;&#x30B1;&#x30FC;&#x30EA;&#x30F3;&#x30B0;&#x306E;&#x9593;&#x306B;&#x826F;&#x597D;&#x306A;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x306B;DNA&#x65AD;&#x7247;&#x30B5;&#x30A4;&#x30BA;&#x306B;&#x3088;&#x308A;&#x3068;&#x5171;&#x306B;&#x5897;&#x52A0;&#x3068;&#x62E1;&#x6563;&#x306E;&#x91CD;&#x8981;&#x306A;&#x5F79;&#x5272;&#x3092;&#xFF08;&#x307B;&#x3068;&#x3093;&#x3069;&#x5B9A;&#x91CF;&#x5316;&#x3057;&#x306A;&#x3044;&#x30D1;&#x30E9;&#x30E1;&#x30FC;&#x30BF;&#xFF09;&#x3092;&#x30CF;&#x30A4;&#x30E9;&#x30A4;&#x30C8;&#x9A5A;&#x304F;&#x3079;&#x304D;&#x4F53;&#x5236;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x305F;&#x3002;&#x3088;&#x308A;&#x901F;&#x304F;&#x3001;&#x5F93;&#x6765;&#x306E;&#x6A5F;&#x5668;&#x3088;&#x308A;&#x3082;&#x6841;&#x9055;&#x3044;&#x306B;&#x3042;&#x308B;&#x5206;&#x96E2;&#x6642;&#x9593;&#x306E;&#x5B9F;&#x7528;&#x7684;&#x306A;&#x5229;&#x70B9;&#x306B;&#x52A0;&#x3048;&#x3066;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x7814;&#x7A76;&#x306E;&#x7D50;&#x679C;&#x306F;&#x3001;&#x6700;&#x9069;&#x306A;&#x5206;&#x96E2;&#x6761;&#x4EF6;&#x3092;&#x7279;&#x5B9A;&#x3057;&#x3001;DNA&#x306E;&#x96FB;&#x6C17;&#x6CF3;&#x52D5;&#x3092;&#x652F;&#x914D;&#x3059;&#x308B;&#x57FA;&#x672C;&#x7684;&#x306A;&#x7269;&#x7406;&#x904E;&#x7A0B;&#x3092;&#x7406;&#x89E3;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x65B0;&#x305F;&#x306A;&#x6D1E;&#x5BDF;&#x3092;&#x8CA2;&#x732E;&#x3059;&#x308B;&#x4EE5;&#x524D;&#x306B;&#x5229;&#x7528;&#x3067;&#x304D;&#x306A;&#x3044;&#x5408;&#x7406;&#x7684;&#x306A;&#x57FA;&#x790E;&#x3092;&#x63D0;&#x4F9B;&#x3059;&#x308B;&#x30D1;&#x30EB;&#x30B9;&#x30D5;&#x30A3;&#x30FC;&#x30EB;&#x30C9;&#x306B;&#x5165;&#x529B;&#x3057;&#x307E;&#x3059;&#x3002;

&#x30EA;&#x30A2;&#x30EB;&#x30BF;&#x30A4;&#x30E0;&#x5168;&#x30B2;&#x30EB;&#x691C;&#x51FA;&#x5668;&#x306B;&#x3088;&#x308B;DNA&#x306E;&#x8FC5;&#x901F;&#x306A;&#x30D5;&#x30A3;&#x30FC;&#x30EB;&#x30C9;&#x53CD;&#x8EE2;&#x30B2;&#x30EB;&#x96FB;&#x6C17;&#x6CF3;&#x52D5;&#x7528;&#x5C0F;&#x578B;&#x30B7;&#x30B9;&#x30C6;&#x30E0;

&#xD384;&#xC2A4; &#xD544;&#xB4DC; &#xC824; &#xC804;&#xAE30; &#xC774;&#xB3D9; &#xBC95; (PFGE) &#xBC29;&#xBC95;&#xC744; &#xB2E4;&#xC591; &#xD55C; DNA &#xBD84;&#xC11D; &#xC751;&#xC6A9; &#xD504;&#xB85C;&#xADF8;&#xB7A8;&#xC5D0; &#xD45C;&#xC900; &#xB3C4;&#xAD6C; &#xB418;&#xACE0;&#xC788;&#xB2E4;. &#xD558;&#xC9C0;&#xB9CC; &#xD384;&#xC2A4; &#xD544;&#xB4DC; &#xC870;&#xAC74; &#xD558;&#xC5D0;&#xC11C; DNA &#xB9C8;&#xC774;&#xADF8;&#xB808;&#xC774;&#xC158; &#xD604;&#xC0C1;&#xC758; &#xC5EC;&#xB7EC; &#xCE21;&#xBA74;&#xC5D0; &#xBE44;&#xD574; &#xC804;&#xAE30; &#xBD84;&#xC57C; &#xC77C;&#xC815; &#xD558; &#xAC8C; &#xC720;&#xC9C0;&#xB294; &#xB354; &#xB9CE;&#xC740; &#xAE30;&#xC874;&#xC758; &#xC0C1;&#xD669; &#xC798; &#xC774;&#xD574; &#xB418;&#xC9C0; &#xC54A;&#xB294;&#xB2E4;. &#xC774; &#xACB0;&#xD54D;&#xC5D0; &#xB300; &#xD55C; &#xC8FC;&#xC694; &#xC774;&#xC720; PFGE &#xC2E4;&#xD5D8;&#xC740; &#xC2DC;&#xAC04; (&#xC57D; 10-15 h) &#xB9E4;&#xC6B0; &#xAE34; &#xBD84;&#xB9AC; &#xBC0F; &#xC2E4;&#xD589; &#xC644;&#xB8CC; &#xB41C; &#xD6C4; poststaining&#xC5D0; &#xC758;&#xD574; &#xC824; &#xBD84;&#xC11D;&#xC744; &#xC218;&#xD589; &#xD558;&#xB294; &#xD544;&#xC694;&#xB97C; &#xC218;&#xD589; &#xD558;&#xB294; &#xC131;&#xAC00;&#xC2E0;&#xC785;&#xB2C8;&#xB2E4;. &#xC27D;&#xAC8C; &#xAD6C;&#xCD95;&#xD560; &#xC18C;&#xD615;&#xB41C; &#xC2AC; &#xB798; &#xBE0C; &#xC824; &#xD070; DNA&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC774;&#xB7EC;&#xD55C; &#xBB38;&#xC81C;&#xB97C; &#xD574;&#xACB0; &#xD558;&#xB294; &#xC7A5;&#xCE58; &#xC870;&#xAC01; &#xCD5C;&#xB300; 35 kb &#xAE38;&#xC774; 60-90 &#xBD84;&#xC5D0; &#xD544;&#xB4DC; &#xBC18;&#xC804; &#xC824; &#xC804;&#xAE30; &#xC774;&#xB3D9; &#xBC95; (FIGE)&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xAD6C;&#xBD84;&#xC744; &#xC18C;&#xAC1C; &#xD558;&#xACA0;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC18C;&#xC790;&#xC758; &#xC18C;&#xD615; &#xD06C;&#xAE30; &#xC804;&#xCCB4; &#xC824;&#xC744; &#xBD84;&#xB9AC; &#xD504;&#xB85C;&#xC138;&#xC2A4; &#xACE0;&#xD574;&#xC0C1;&#xB3C4; CCD &#xCE74;&#xBA54;&#xB77C;&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC2E4;&#xC2DC;&#xAC04;&#xC73C;&#xB85C;&#xC5D0;&#xC11C; &#xC774;&#xBBF8;&#xC9C0;&#xB97C; &#xB9CC;&#xB4E4; &#xC218; &#xC788;&#xB3C4;&#xB85D; &#xC9C0;&#xC18D;&#xC801;&#xC73C;&#xB85C; &#xBAA8;&#xB2C8;&#xD130;&#xB9C1;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC784;&#xC758; &#xC801;&#xC6A9;&#xB41C; &#xC804;&#xC555; &#xD30C;&#xD615; &#xC81C;&#xC5B4; &#xD568;&#xC218; &#xBC1C;&#xC0DD;&#xAE30;&#xB294; &#xACE0;&#xC804;&#xC555; &#xC99D;&#xD3ED;&#xAE30;&#xC640; &#xC778;&#xD130;&#xD398;&#xC774;&#xC2A4;&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC5BB;&#xC744; &#xC218; &#xC788;&#xB2E4;. &#xC774;&#xB7EC;&#xD55C; &#xAE30;&#xB2A5;&#xC740; DNA &#xB9C8;&#xC774;&#xADF8;&#xB808;&#xC774;&#xC158; (&#xC131;&#xACFC; &#xD655;&#xC0B0;, &#xBD84;&#xB9AC; &#xD574;&#xC0C1;&#xB3C4;)&#xC5D0; &#xAD00;&#xB828; &#xB41C; &#xAE30;&#xBCF8;&#xC801;&#xC778; &#xBB3C;&#xB9AC;&#xC801; &#xB9E4;&#xAC1C;&#xC758; &#xD06C;&#xAE30; &#xC758;&#xC874;&#xC131;&#xC744; &#xC870;&#xC0AC;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC774;&#xB7EC;&#xD55C; &#xB370;&#xC774;&#xD130; &#xB180;&#xB77C;&#xC6B4; &#xC815;&#xAD8C;&#xC774; &#xC5B4;&#xB514; &#xBD84;&#xB9AC; &#xD574;&#xC0C1;&#xB3C4; DNA &#xC870;&#xAC01; &#xD06C;&#xAE30; &#xB54C;&#xBB38;&#xC5D0; &#xC774;&#xB3D9; &#xC131;&#xACFC; &#xD655;&#xC0B0; &#xC0AC;&#xC774; &#xC720;&#xB9AC;&#xD55C; &#xC0C1;&#xD638; &#xC791;&#xC6A9;&#xC73C;&#xB85C; &#xC81C;&#xC870;&#xC2DC; &#xC99D;&#xAC00; &#xACF5;&#xAC1C; &#xBC0F; &#xBCF4;&#xAE09; (&#xB4DC;&#xBB3C;&#xAC8C; &#xC815;&#xB7C9;&#xB41C; &#xB9E4;&#xAC1C; &#xBCC0;&#xC218;)&#xC758; &#xC911;&#xC694; &#xD55C; &#xC5ED;&#xD560;&#xC744; &#xAC15;&#xC870;. &#xBD84;&#xB9AC; &#xC2DC;&#xAC04;&#xC744; &#xD06C;&#xAE30; &#xC21C;&#xC11C; &#xAE30;&#xC874;&#xC758; &#xC7A5;&#xBE44; &#xBCF4;&#xB2E4; &#xB354; &#xBE68;&#xB9AC; &#xC2E4;&#xC9C8;&#xC801;&#xC778; &#xD61C;&#xD0DD; &#xBFD0;&#xB9CC; &#xC544;&#xB2C8;&#xB77C; &#xC774;&#xB7EC;&#xD55C; &#xC5F0;&#xAD6C;&#xC758; &#xACB0;&#xACFC; &#xCD5C;&#xC801; &#xBD84;&#xB9AC; &#xC870;&#xAC74;&#xC744; &#xC2DD;&#xBCC4; &#xD558; &#xACE0; &#xD384;&#xC2A4; &#xD544;&#xB4DC;&#xC5D0;&#xC11C; DNA &#xC804;&#xAE30; &#xC774;&#xB3D9; &#xBC95;&#xC744; &#xC81C;&#xC5B4; &#xD558;&#xB294; &#xAE30;&#xBCF8; &#xC2E4;&#xC81C; &#xD504;&#xB85C;&#xC138;&#xC2A4; &#xC774;&#xD574;&#xB97C; &#xD5A5;&#xD574; &#xC0C8;&#xB85C;&#xC6B4; &#xD1B5;&#xCC30;&#xB825;&#xC744; &#xAE30;&#xC5EC; &#xD558; &#xC774;&#xC804;&#xC5D0; &#xC0AC;&#xC6A9;&#xD560; &#xC218; &#xD569;&#xB9AC;&#xC801;&#xC778; &#xADFC;&#xAC70;&#xB97C; &#xC81C;&#xACF5;.

&#xBE60;&#xB978; &#xD544;&#xB4DC; &#xBC18;&#xC804; &#xC704;&#xD55C; &#xC18C;&#xD615;&#xB41C; &#xC2DC;&#xC2A4;&#xD15C; &#xC824; &#xC2E4;&#xC2DC;&#xAC04; &#xC804;&#xCCB4; &#xC824; &#xAC80;&#xCD9C; dna &#xC804;&#xAE30; &#xC774;&#xB3D9; &#xBC95;.

Pulsatorios de electroforesis en gel de campo (PFGE) m&#xE9;todos se han convertido en herramientas est&#xE1;ndar en una amplia gama de aplicaciones de an&#xE1;lisis de ADN. Pero muchos aspectos de los fen&#xF3;menos de migraci&#xF3;n de ADN en condiciones de campo de impulsos no se conocen bien en comparaci&#xF3;n con la situaci&#xF3;n m&#xE1;s convencional, donde el campo el&#xE9;ctrico se mantiene constante. Una raz&#xF3;n clave para esta deficiencia es que los experimentos PFGE son engorrosos de realizar debido a los tiempos de separaci&#xF3;n extremadamente largo (aproximadamente 10-15 h) y la necesidad de realizar an&#xE1;lisis en gel por poststaining despu&#xE9;s de la terminaci&#xF3;n de la carrera. Aqu&#xED; te presentamos una forma f&#xE1;cil de crear un aparato miniaturizado gel de losa que se ocupa de estas cuestiones, permitiendo grandes fragmentos de ADN de hasta 35 kb de longitud que se separan con inversi&#xF3;n de campo en la electroforesis en gel (FIGE) en 60-90 min. El tama&#xF1;o compacto del dispositivo tambi&#xE9;n permite que el gel entero para controlar continuamente de modo que los procesos de separaci&#xF3;n se pueden obtener im&#xE1;genes en tiempo real utilizando una c&#xE1;mara CCD de alta resoluci&#xF3;n. Control arbitrario sobre las formas de onda de tensi&#xF3;n aplicada se consigue utilizando un generador de funciones en interfase con un amplificador de alta tensi&#xF3;n. Estas capacidades nos permiten investigar la dependencia del tama&#xF1;o de los fundamentales par&#xE1;metros f&#xED;sicos asociados con el ADN de la migraci&#xF3;n (movilidad, difusi&#xF3;n, y la resoluci&#xF3;n de separaci&#xF3;n). Estos datos revelan un r&#xE9;gimen de extra&#xF1;ar que los aumentos de separaci&#xF3;n de resoluci&#xF3;n con el tama&#xF1;o de fragmentos de ADN, debido a una interacci&#xF3;n favorable entre la movilidad y batiduras de difusi&#xF3;n y poner de relieve el importante papel de difusi&#xF3;n (un par&#xE1;metro rara vez cuantificada). Adem&#xE1;s de la utilidad pr&#xE1;ctica de los tiempos de separaci&#xF3;n que son un orden de magnitud m&#xE1;s r&#xE1;pido que los instrumentos convencionales, los resultados de estos estudios proporcionan una base racional antes no estaban disponibles para identificar las condiciones &#xF3;ptimas de separaci&#xF3;n y contribuir a la comprensi&#xF3;n de nuevos conocimientos de los procesos f&#xED;sicos subyacentes que gobiernan la electroforesis de ADN en los campos pulsados.

Sistema miniaturizado de electroforesis en gel de campo de inversi&#xF3;n r&#xE1;pida del ADN en tiempo real con la Totalidad del gel de detecci&#xF3;n

Heart failure is a debilitating condition resulting in severe disability and death. In a subset of cases, clustered as idiopathic dilated cardiomyopathy (iDCM), the origin of heart failure is unknown. In the brain of patients with dementia, proteinaceous aggregates and abnormal oligomeric assemblies of beta-amyloid impair cell function and lead to cell death.

Protein aggregates and novel presenilin gene variants in idiopathic dilated cardiomyopathy.

The SRY-related HMG-box 5 (SOX5) gene encodes a member of the SOX family of transcription factors. Recently, genome-wide association studies have implicated SOX5 as a candidate gene for susceptibility to four cardiac-related endophenotypes: higher resting heart rate (HR), the electrocardiographic PR interval, atrial fibrillation and left ventricular mass. We have determined that human SOX5 has a highly conserved Drosophila ortholog, Sox102F, and have employed transgenic Drosophila models to quantitatively measure cardiac function in adult flies. For this purpose, we have developed a high-speed and ultrahigh-resolution optical coherence tomography imaging system, which enables rapid cross-sectional imaging of the heart tube over various cardiac cycles for the measurement of cardiac structural and dynamical parameters such as HR, dimensions and areas of heart chambers, cardiac wall thickness and wall velocities. We have found that the silencing of Sox102F resulted in a significant decrease in HR, heart chamber size and cardiac wall velocities, and a significant increase in cardiac wall thickness that was accompanied by disrupted myofibril structure in adult flies. In addition, the silencing of Sox102F in the wing led to increased L2, L3 and wing marginal veins and increased and disorganized expression of wingless, the central component of the Wnt signaling pathway. Collectively, the silencing of Sox102F resulted in severe cardiac dysfunction and structural defects with disrupted Wnt signaling transduction in flies. This implicates an important functional role for SOX5 in heart and suggests that the alterations in SOX5 levels may contribute to the pathogenesis of multiple cardiac diseases or traits.

Silencing of the Drosophila ortholog of SOX5 in heart leads to cardiac dysfunction as detected by optical coherence tomography.

Ceriporia lacerata, a strain of white-rot fungus isolated from the litter of an invasive plant (Solidago canadensis) in China, was little known about its properties and utilization. In this work, the copper(II) biosorption characteristics of formaldehyde inactivated C. lacerata biomass were examined as a function of initial pH, initial copper(II) concentration and contact time, and the adsorptive equilibrium and kinetics were simulated, too.

Equilibrium and kinetic studies of copper biosorption by dead Ceriporia lacerata biomass isolated from the litter of an invasive plant in China.

Circadian rhythms are endogenous, entrainable oscillations of physical, mental and behavioural processes in response to local environmental cues such as daylight, which are present in the living beings, including humans. Circadian rhythms have been related to cardiovascular function and pathology. However, the role that circadian clock genes play in heart development and function in a whole animal in vivo are poorly understood. The Drosophila cryptochrome (dCry) is a circadian clock gene that encodes a major component of the circadian clock negative feedback loop. Compared to the embryonic stage, the relative expression levels of dCry showed a significant increase (>100-fold) in Drosophila during the pupa and adult stages. In this study, we utilized an ultrahigh resolution optical coherence microscopy (OCM) system to perform non-invasive and longitudinal analysis of functional and morphological changes in the Drosophila heart throughout its post-embryonic lifecycle for the first time. The Drosophila heart exhibited major morphological and functional alterations during its development. Notably, heart rate (HR) and cardiac activity period (CAP) of Drosophila showed significant variations during the pupa stage, when heart remodeling took place. From the M-mode (2D + time) OCM images, cardiac structural and functional parameters of Drosophila at different developmental stages were quantitatively determined. In order to study the functional role of dCry on Drosophila heart development, we silenced dCry by RNAi in the Drosophila heart and mesoderm, and quantitatively measured heart morphology and function in those flies throughout its development. Silencing of dCry resulted in slower HR, reduced CAP, smaller heart chamber size, pupal lethality and disrupted posterior segmentation that was related to increased expression of a posterior compartment protein, wingless. Collectively, our studies provided novel evidence that the circadian clock gene, dCry, plays an essential role in heart morphogenesis and function.

A Circadian Clock Gene, Cry, Affects Heart Morphogenesis and Function in Drosophila as Revealed by Optical Coherence Microscopy.

Electrical stimulation is currently the gold standard for cardiac pacing. However, it is invasive and nonspecific for cardiac tissues. We recently developed a noninvasive cardiac pacing technique using optogenetic tools, which are widely used in neuroscience. Optogenetic pacing of the heart provides high spatial and temporal precisions, is specific for cardiac tissues, avoids artifacts associated with electrical stimulation, and therefore promises to be a powerful tool in basic cardiac research. We demonstrated optogenetic control of heart rhythm in a well-established model organism, Drosophila melanogaster. We developed transgenic flies expressing a light-gated cation channel, channelrhodopsin-2 (ChR2), specifically in their hearts and demonstrated successful optogenetic pacing of ChR2-expressing Drosophila at different developmental stages, including the larva, pupa, and adult stages. A high-speed and ultrahigh-resolution optical coherence microscopy imaging system that is capable of providing images at a rate of 130 frames/s with axial and transverse resolutions of 1.5 and 3.9 μm, respectively, was used to noninvasively monitor Drosophila cardiac function and its response to pacing stimulation. The development of a noninvasive integrated optical pacing and imaging system provides a novel platform for performing research studies in developmental cardiology.

Optogenetic pacing in Drosophila melanogaster.

Since both ethanol and butanol fermentations are urgently developed processes with the biofuel-demand increasing, performance comparison of aerobic ethanol fermentation and anerobic butanol fermentation in a continuous and closed-circulating fermentation (CCCF) system was necessary to achieve their fermentation characteristics and further optimize the fermentation process. Fermentation and pervaporation parameters including the average cell concentration, glucose consumption rate, cumulated production concentration, product flux, and separation factor of ethanol fermentation were 11.45 g/L, 3.70 g/L/h, 655.83 g/L, 378.5 g/m(2)/h, and 4.83, respectively, the corresponding parameters of butanol fermentation were 2.19 g/L, 0.61 g/L/h, 28.03 g/L, 58.56 g/m(2)/h, and 10.62, respectively. Profiles of fermentation and pervaporation parameters indicated that the intensity and efficiency of ethanol fermentation was higher than butanol fermentation, but the stability of butanol fermentation was superior to ethanol fermentation. Although the two fermentation processes had different features, the performance indicated the application prospect of both ethanol and butanol production by the CCCF system. Abbreviation: CCCF, continuous and closed-circulating fermentation.

Performance comparison of ethanol and butanol production in a continuous and closed-circulating fermentation system with membrane bioreactor.

Optical coherence tomography (OCT) is a promising research tool for brain imaging and developmental biology. Serving as a three-dimensional optical biopsy technique, OCT provides volumetric reconstruction of brain tissues and embryonic structures with micrometer resolution and video rate imaging speed. Functional OCT enables label-free monitoring of hemodynamic and metabolic changes in the brain in vitro and in vivo in animal models. Due to its non-invasiveness nature, OCT enables longitudinal imaging of developing specimens in vivo without potential damage from surgical operation, tissue fixation and processing, and staining with exogenous contrast agents. In this paper, various OCT applications in brain imaging and developmental biology are reviewed, with a particular focus on imaging heart development. In addition, we report findings on the effects of a circadian gene (Clock) and high-fat-diet on heart development in Drosophila melanogaster. These findings contribute to our understanding of the fundamental mechanisms connecting circadian genes and obesity to heart development and cardiac diseases.

ArticleTotal : 2	Year
Publication title	2016
Publication title	2019

Article	Year
Clinical profile of autosomal dominant polycystic liver disease. Hepatology (Baltimore, Md.)\| PubMed ID: 12500201	2003
Mutations in PRKCSH cause isolated autosomal dominant polycystic liver disease. American journal of human genetics\| PubMed ID: 12529853	2003
Identification of two novel polycystic kidney disease-1-like genes in human and mouse genomes. Genomics\| PubMed ID: 12782129	2003
Mutations in SEC63 cause autosomal dominant polycystic liver disease. Nature genetics\| PubMed ID: 15133510	2004
Isolation and characterization of the Drosophila ubiquilin ortholog dUbqln: in vivo interaction with early-onset Alzheimer disease genes. Human molecular genetics\| PubMed ID: 17704509	2007
Miniaturized system for rapid field inversion gel electrophoresis of DNA with real-time whole-gel detection. Analytical chemistry\| PubMed ID: 20148578	2010
Protein aggregates and novel presenilin gene variants in idiopathic dilated cardiomyopathy. Circulation\| PubMed ID: 20194882	2010
Silencing of the Drosophila ortholog of SOX5 in heart leads to cardiac dysfunction as detected by optical coherence tomography. Human molecular genetics\| PubMed ID: 23696452	2013
Equilibrium and kinetic studies of copper biosorption by dead Ceriporia lacerata biomass isolated from the litter of an invasive plant in China. Journal of environmental health science & engineering\| PubMed ID: 25954509	2015
A Circadian Clock Gene, Cry, Affects Heart Morphogenesis and Function in Drosophila as Revealed by Optical Coherence Microscopy. PloS one\| PubMed ID: 26348211	2015
Optogenetic pacing in Drosophila melanogaster. Science advances\| PubMed ID: 26601299	2015
Performance comparison of ethanol and butanol production in a continuous and closed-circulating fermentation system with membrane bioreactor. Preparative biochemistry & biotechnology\| PubMed ID: 27558450	2016
Optical Coherence Tomography for Brain Imaging and Developmental Biology. IEEE journal of selected topics in quantum electronics : a publication of the IEEE Lasers and Electro-optics Society\| PubMed ID: 27721647	2016

Airong Li

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