Gli autori ringraziano l&#39;odontotecnico Nicola Lanfranconi per produrre il modello di riferimento principale e la Società Alicona per il loro supporto continuo al miglioramento del software di scansione.

1. Preparazione del campione / Base <ol><li> Applicare una base piatta intorno al provino. Posizionare il campione sul tavolo di scansione. Orientate la superficie occlusale sul piano orizzontale (Figura 1). </li></ol> 2. Analysis Software <ol><li> Avviare il programma e poi il Laboratorio modulo di misura (figura 2) </li><li> Posizionare il campione al centro del tavolo di scansione. </li><li> Scegliere la lente di ingrandimento corretta. Per oggetti di grandi dimensioni, come scansioni complete arco, utilizzare l&#39;obiettivo 5X. </li><li> Spostare la scansione ottica utilizzando il mouse 3D fino a visualizzare la superficie del campione sulla finestra di visualizzazione in tempo reale (Figura 3). </li><li> Utilizzare il controllo sensore per impostare esposizione e il contrasto per ottenere parametri di scansione ottimali. Per superfici metalliche, utilizzare un esposizione tra 400 e 800 msec e un contrasto tra 0,3 e 0,8 (Figura 4). </li><li> Controllare la qualità dell&#39;immagine selezionando il pulsante&quot;Mostra qualità delle immagini in Live Preview&quot;. </li><li> Impostare i parametri corretti nella sezione &quot;Controllo della misura&quot;. Il tipo di misura è &quot;3D Dataset&quot;; ImageField Type è &quot;General ImageField&quot; (Figura 5). </li><li> Fare clic su &quot;Nuovo ImageField&quot;. </li><li> Definire il volume di scansione. Il software deve le coordinate di delimitazione del volume di misura. <ol><li> Spostare il campione al livello più alto e più basso scansione e fare clic su &quot;Aggiungi Position&quot; in entrambi i punti. Il valore &quot;Z Range&quot; indica l&#39;altezza effettiva del volume di scansione (Figura 6). </li><li> Spostare il campione ai boundings XY del volume di scansione. Fare clic su &quot;Aggiungi Position&quot; per definire l&#39;asse X e lunghezze y. La dimensione del volume di scansione viene visualizzato nella scheda &quot;Informazioni&quot; e deve superare le dimensioni del campione da 1 cm l&#39;asse X e Y. </li></ol></li><li> Controllare il numero di &quot;punti&quot;. Il software è in grado di scansione di 100 milioni di punti di superficie in una sola scansione &quot;ImageField&quot;. Il numero effettivodi punti supera questo limite. &quot;Decimate&quot; il numero del punto di misura da un &quot;downsampling laterale&quot; della dimensione in punti. <ol><li> Clicca su &quot;Impostazioni avanzate&quot; (Figura 7). </li><li> Spostare il cursore &quot;Lateral Downsampling&quot; a destra fino a quando il numero di &quot;punti&quot; si riduce al di sotto di 100 M (Figura 7). Downsampling riduce la risoluzione laterale dei punti della superficie, con conseguente grande dimensione in pixel dell&#39;oggetto esaminato. Per garantire risultati di scansione ottimali, la risoluzione verticale è aumentata. </li></ol></li><li> Fare clic su &quot;Start misura&quot;. Verrà avviata la &quot;modalità di anteprima&quot;. Il software esegue una scansione preliminare con le dimensioni X e Y assi selezionati. </li><li> Dopo la scansione preliminare è completata, selezionare la regione di interesse. Questo aiuta a ridurre le dimensioni del file e il tempo di scansione (Figura 8). </li><li> Selezionare tutte le parti della scansione preliminare eccezione del campione e due campi di misura intorno al provino, contenente la base piatta (Figura 9). </li><li> Fare clic su &quot;Start&quot; per avviare la scansione. </li><li> Controllare la scansione in &quot;ImageViewer&quot; con il mouse e premere il tasto sinistro del mouse (Figura 10). </li><li> Chiudere la finestra e fare clic su &quot;Visualizza Pseudo Color Only&quot;. </li><li> Clicca su &quot;Impostazioni&quot; e &quot;Pseudo Colorare&quot;, e scegliere &quot;ripetibilità&quot;. </li><li> Impostare il &quot;Max&quot;. valore di 0,2 e clicca su &quot;Applica intervallo&quot;. </li><li> Controllare la ripetibilità, dovrebbe essere uguale per aree con la stessa inclinazione e materiale. In particolare, la base intorno il campione dovrebbe avere una ripetibilità omogenea (Figura 11). </li><li> Clicca su &quot;Database&quot; e salvare la scansione nella cartella appropriata. </li><li> Esportare la scansione di diversi formati di file, se necessario. Clicca su &quot;File/Export/3D Dati / ....&quot; Il protocollo può essere messo in pausa in quel punto e continuò più tardi. </li></ol> 3. Analisi Differenza <ol><li> Utilizzare il &quot;3D-Editor&quot; per tagliare via la base. Quest&#39;area non sarà utilizzata per la differenzaanalisi. </li><li> Per confrontare e analizzare i due scansioni, avviare il software &quot;Misuratore della differenza&quot; (Figura 12). </li><li> Scegliere il secondo modello da confrontare. </li><li> Clicca su &quot;Automatico Ruvido Allineamento&quot; per effettuare una prima partita dei modelli. </li><li> Clicca su &quot;Regolazione manuale&quot; e allineare i modelli con rotazione e lo spostamento fino a quando sono nello stesso orientamento (Figura 13). </li><li> Clicca su &quot;allineamento automatico&quot; e &quot;Applica&quot; per avviare il miglior algoritmo adatto per corrispondenza di modello ottimale. </li><li> Clicca su &quot;Differenze&quot; per visualizzare la mappa differenza dei due modelli abbinati. </li><li> Selezionare un intervallo di colori corretta per visualizzare le deviazioni (Figura 14). Salvare il modello deviazione visiva come uno schermo per l&#39;analisi. </li></ol> Clicca su &quot;Statistica&quot; per visualizzare i valori statistici delle differenze. Scegli la dimensione della classe 1 micron e salvare i dati di istogramma in un file di testo per il confronto statistico (Figura 15).

<ol>
	<li>Wettstein, F., Sailer, I., Roos, M., H&#228;mmerle, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+study+of+the+internal+gaps+of+zirconia+and+metal+frameworks+for+fixed+partial+dentures.">Clinical study of the internal gaps of zirconia and metal frameworks for fixed partial dentures.</a> Eur. J. Oral Sci. 116 (3), 272-279 (2008).</li><li>Persson, A. S., Oden, A., Andersson, M., Sandborgh-Englund, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Digitization+of+simulated+clinical+dental+impressions:+virtual+three-dimensional+analysis+of+exactness.">Digitization of simulated clinical dental impressions: virtual three-dimensional analysis of exactness.</a> Dent. Mater. 25 (7), 929-936 (2009).</li><li>Del'Acqua, M. A., Arioli-Filho, J. N., Compagnoni, M. A., Mollo, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=de+A+Accuracy+of+impression+and+pouring+techniques+for+an+implant-supported+prosthesis.">de A Accuracy of impression and pouring techniques for an implant-supported prosthesis.</a> Int. J. Oral Maxillofac. Implants. 23 (2), 226-236 (2008).</li><li>Schneider, D., Grunder, U., Ender, A., H&#228;mmerle, C. H. F., Jung, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volume+gain+and+stability+of+peri-implant+tissue+following+bone+and+soft+tissue+augmentation:+1-year+results+from+a+prospective+cohort+study.">Volume gain and stability of peri-implant tissue following bone and soft tissue augmentation: 1-year results from a prospective cohort study.</a> Clinical Oral Implants Research. 22 (1), 28-37 (2011).</li><li>Windisch, S. I., Jung, R. E., Sailer, I., Studer, S. P., Ender, A., H&#228;mmerle, C. H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+optical+method+to+evaluate+three-dimensional+volume+changes+of+alveolar+contours:+a+methodological+in+vitro+study.">A new optical method to evaluate three-dimensional volume changes of alveolar contours: a methodological in vitro study.</a> Clinical Oral Implants Research. 18 (5), 545-551 (2007).</li><li>Caputi, S., Varvara, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dimensional+accuracy+of+resultant+casts+made+by+a+monophase,+one-step+and+two-step,+and+a+novel+two-step+putty/light-body+impression+technique:+an+in+vitro.">Dimensional accuracy of resultant casts made by a monophase, one-step and two-step, and a novel two-step putty/light-body impression technique: an in vitro.</a> 99 (4), 274-281 (2008).</li><li>Hoyos, A., Soderholm, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+tray+rigidity+and+impression+technique+on+accuracy+of+polyvinyl+siloxane+impressions.">Influence of tray rigidity and impression technique on accuracy of polyvinyl siloxane impressions.</a> Int. J. Prosthodont. 24 (1), 49-54 (2011).</li><li>Luthardt, R. G., Kuhmstedt, P., Walter, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+method+for+the+computer-aided+evaluation+of+three-dimensional+changes+in+gypsum+materials.">A new method for the computer-aided evaluation of three-dimensional changes in gypsum materials.</a> Dent. Mater. 19 (1), 19-24 (2003).</li><li>Mehl, A., Ender, A., M&#246;rmann, W., Attin, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+testing+of+a+new+intraoral+3D+camera.">Accuracy testing of a new intraoral 3D camera.</a> International Journal of Computerized Dentistry. 12 (1), 11-28 (2009).</li><li>Fickl, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dimensional+changes+of+the+ridge+contour+after+socket+preservation+and+buccal+overbuilding:+an+animal+study.">Dimensional changes of the ridge contour after socket preservation and buccal overbuilding: an animal study.</a> J. Clin. Periodontol. 36 (5), 442-448 (2009).</li><li>Vlaar, S. T., vander Zel, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+of+dental+digitizers.">Accuracy of dental digitizers.</a> Int. Dent. J. 56 (5), 301-309 (2006).</li><li>Ender, A., Mehl, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+of+complete-arch+dental+impressions:+a+new+method+of+measuring+trueness+and+precision.">Accuracy of complete-arch dental impressions: a new method of measuring trueness and precision.</a> 109 (2), 121-128 (2013).</li><li>Ender, A., Mehl, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+scanning+strategies+on+the+accuracy+of+digital+intraoral+scanning+systems.">Influence of scanning strategies on the accuracy of digital intraoral scanning systems.</a> Int. J. Comput. Dent. 16 (1), 11-21 (2013).</li><li>Ender, A., Mehl, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Full+arch+scans:+conventional+versus+digital+impressions--an+in-vitro+study.">Full arch scans: conventional versus digital impressions--an in-vitro study.</a> Int. J. Comput. Dent. 14 (1), 11-21 (2011).</li><li>Meer, W. J., Andriessen, F. S., Wismeijer, D., Ren, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+intra-oral+dental+scanners+in+the+digital+workflow+of+implantology.">Application of intra-oral dental scanners in the digital workflow of implantology.</a> PLoS One. 7 (8), (2012).</li></ol>

Gli autori dichiarano di non avere interessi finanziari concorrenti o altri conflitti di interesse.

La precisione è una richiesta fondamentale nella medicina dentale. Lo scanner di riferimento è in grado di scansionare oggetti piccoli e grandi, con alta esattezza e precisione. Con il metodo di scansione ottimale, anche le singole superfici dentali dettagliate morfologici possono essere scansionate ad alta risoluzione e ripetibilità. Con i diversi livelli di ingrandimento dello scanner, strutture morfologiche macro e micro possono essere acquisite. È possibile scansionare una varietà di materiali modello. <p c...

La precisione è un grande interesse in molti campi della medicina dentale. Sostituzione di tessuti duri dentali ha bisogno di una protesi esatta raccordo per assicurare il corretto funzionamento ed evitare di distruggere ulteriormente la restante struttura del dente 1,2. Protesi parziali fisse e protesi totale sono particolarmente critici per il montaggio esatto in strutture di sostegno come i denti preparati o impianti 3. Questo è il motivo per una riproduzione altamente accurata è necessaria, soprattutto nel campo delle impronte dentali e workflow laboratorio odontotecnico. Tuttavia, altri campi di trattamento odontoiatrico beneficiano anche di un vero e preciso risultato metrica, per verificare il successo del trattamento e valutare nuove strategie di trattamento, ad esempio, l&#39;aumento dei tessuti molli e duri, erosione e abrasione monitoraggio, trattamenti parodontali e trattamenti ortodontici 4,5. In molti di questi campi, procedure di validazione attuali sono misure di distanza lineari con pinze o microscopi 6,7. Questi methods sono limitati a pochi punti di misura e informazioni limitato di tridimensionali (3D) il cambiamento di zona di prova. Metodi di misurazione più recenti comprendono il rilevamento ottico o radiografica di tutta la superficie dell&#39;oggetto 8,9 prova. Qui, l&#39;intera superficie o il volume viene misurata e visualizzata come un oggetto 3D sullo schermo del computer. Misure lineari sono possibili, così come sovrapposizioni di modelli da tempi di scansione diverse. Con questa sovrapposizione, una valutazione dei cambiamenti in ogni punto della superficie di scansione è possibile. Ciò consente la possibilità di monitorare una determinata area o la visualizzazione deformazioni in tutti e tre gli assi cartesiani. Inoltre, variazioni volumetriche possono essere misurati 10. Il punto limitare a questi nuovi metodi è la precisione dello scanner, utilizzato per acquisire l&#39;oggetto di prova. Nessuno dei cambiamenti all&#39;interno della precisione dello scanner riferimento può essere diviso in variazioni dell&#39;oggetto della prova o errori di scansione. Precisione Scan è spesso un valore dato dal costruttorecostruttore derivato dalla scansione piccoli oggetti calibrate 11. Questo errore di scansione minimo è diversa quando la scansione di oggetti di grandi dimensioni, come un arco dentale. Accuratezza compone di veridicità e precisione. L&#39;esattezza è la deviazione dell&#39;oggetto scansionato dalla sua reale geometria. La precisione è la deviazione tra le scansioni ripetute (ISO 5725-1). In questo studio, un nuovo scanner ottico di riferimento, sulla base della variazione tecnica di scansione attivo, è stato introdotto per analizzare campioni di singolo dente fino a modelli dell&#39;arcata completa con la massima precisione. Questo scanner di riferimento è stato utilizzato come base per diversi studi, confrontando dentale accuratezza impressione convenzionali e tecniche digitali 12-14 e progetti concreti riguardanti occlusione dentale e all&#39;abrasione dei materiali dentali. Lo scopo di questo studio era quello di fornire informazioni di base della precisione dello scanner riferimento e alcune possibilità di utilizzare questo dispositivo nel campo della ricerca dentale.

<table border="0" cellpadding="0" cellspacing="0" width="669">
	<tbody>
		<tr>
			<td>Reference model</td>
			<td></td>
			<td></td>
			<td>individual non-precious metal model, derived from a patient impression</td>
		</tr>
		<tr>
			<td>Araldit repair</td>
			<td>Huntsmen Advanced Material, Basel, Switzerland</td>
			<td></td>
			<td>used for making the base of the reference model</td>
		</tr>
		<tr>
			<td>CamBase</td>
			<td>Dentona, Dortmund, Germany</td>
			<td></td>
			<td>Type IV dental ston for pouring conventional impressions</td>
		</tr>
		<tr>
			<td>Identium</td>
			<td>Kettenbach, Eschenburg, Germany</td>
			<td></td>
			<td>Vinylsiloxanether impression material for conventional impression</td>
		</tr>
		<tr>
			<td>inEOS model holder</td>
			<td>Sirona Dental Systems, Bensheim, Germany</td>
			<td></td>
			<td>used for fixing stone models at the reference scanner</td>
		</tr>
		<tr>
			<td>Accutrans</td>
			<td>Coltene Whaledent, Altst&#228;tten, Switzerland</td>
			<td></td>
			<td>used for making the base of thestone models</td>
		</tr>
		<tr>
			<td>President putty</td>
			<td>Coltene Whaledent, Altst&#228;tten, Switzerland</td>
			<td></td>
			<td>mix with accutrans for betterstability of the base</td>
		</tr>
		<tr>
			<td>Alicona Infinite Focus</td>
			<td>Alicona Imaging, Graz, Austria</td>
			<td></td>
			<td>Reference scanner&#160;</td>
		</tr>
	</tbody>
</table>

accuracy in dental medicine, a new way to measure trueness and precision

Scanner di riferimento sono utilizzati in odontoiatria per verificare un sacco di procedure. L&#39;interesse principale è quello di verificare i metodi impressione che servono come base per le protesi dentarie. L&#39;attuale limitazione di molti scanner di riferimento è la mancanza di scansione precisione oggetti di grandi dimensioni come arcate dentali complete, o la limitata possibilità di valutare superfici dettagliate denti. Un nuovo scanner di riferimento, sulla base di messa a fuoco variazione tecnica di scansione, è stata valutata per quanto riguarda la massima precisione locale e generale. Un protocollo di scansione specifico è stato testato per la scansione di superficie originale dei denti da impronte dentali. Inoltre, diversi materiali di modello sono stati verificati. I risultati hanno mostrato una precisione di scansione ad alta dello scanner di riferimento con una deviazione media di 5,3 ± 1,1 micron per la veridicità e 1,6 ± 0,6 micron per la precisione nel caso di scansioni complete arco. Metodi di impronte dentali correnti evidenziano scostamenti molto più alto (esattezza: 20.4 ± 2.2 micron, precisione: 12,5 ± 2,5 micron) than l&#39;accuratezza scansione interna dello scanner di riferimento. Oggetti più piccoli come un&#39;unica superficie del dente possono essere acquisiti con una precisione ancora più elevata, consentendo al sistema di valutare erosiva e abrasiva perdita di superficie del dente. Lo scanner riferimento può essere utilizzato per misurare le differenze per un sacco di campi di ricerca dentale. I diversi livelli di ingrandimento in combinazione con una elevata precisione locale e generale possono essere utilizzati per valutare i cambiamenti di denti singoli o restauri fino ai cambiamenti completi arch.

La figura 16A mostra la scansione di un campione di gesso. Per verificare i parametri di scansione ottimali, controllare la ripetibilità del materiale. Una sezione del campione viene scansionato con diverse impostazioni di contrasto e luminosità e la ripetibilità viene controllato dopo ogni scansione per trovare le impostazioni di scansione ottimali. Le superfici metalliche mostrano i migliori risultati con un contrasto minore 1.0 e gesso o resina materiali con un contrasto maggiore. Figura 1...

Watch this Scientific Journal Video about Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision at JoVE.com

Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision

La precisione è una grande domanda in odontoiatria. Per verificare la precisione, sono necessari gli scanner di riferimento. Questo articolo presenta un nuovo scanner di riferimento, con un metodo di scansione adjusted di acquisire una vasta gamma di morfologie dentali con elevata esattezza e precisione.

Accuratezza nella Dental Medicine, Un nuovo modo di misurare veridicità e precisione

Reference scanners are used in dental medicine to verify a lot of procedures. The main interest is to verify impression methods as they serve as a base ...

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Research

JoVE Journal

Medicine

13.3K Views.  University of Zürich.  La precisione è una grande domanda in odontoiatria. Per verificare la precisione, sono necessari gli scanner di riferimento. Questo articolo presenta un nuovo scanner di riferimento, con un metodo di scansione adjusted di acquisire una vasta gamma di morfologie dentali con elevata esattezza e precisione. 

Video: Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision

Technique to Collect Fungiform (Taste) Papillae from Human Tongue

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic processes that prepare the alimentary canal for digestion. Steady progress is being made towards understanding the early biochemical and molecular events underlying taste transduction (for a review, Breslin and Spector, 20081). However, progress to date has largely resulted from animal models. Yet, since marked differences in receptor specificity and receptor density vary among species, human taste transduction will only be understood by using human taste tissue. Here we describe a biopsy technique to collect human fungiform papillae, visible as rounded pink anterior structures, about 0.5 mm in diameter that contain taste buds. These biopsied papillae are used for several purposes including the isolation of viable taste bud cells, in situ hybridization, immunohistochemistry and, through techniques of molecular biology, the identification of taste-specific novel proteins.

Technique to Collect Fungiform Taste Papillae from Human Tongue

Knowledge of molecular mechanisms underlying gustatory transduction has recently enjoyed significant advances, largely due to using animal models. However, the wide diversity in taste sensitivity and specificity among mammals warrants studies in human tissue. We describe a biopsy technique to collect living taste cells from the papillae on human tongue.

Tecnica per raccogliere fungiformi (Gusto) Papille da lingua umana

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic ...

Ricerca

Medicina

Construction of Vapor Chambers Used to Expose Mice to Alcohol During the Equivalent of all Three Trimesters of Human Development

Exposure to alcohol during development can result in a constellation of morphological and behavioral abnormalities that are collectively known as Fetal Alcohol Spectrum Disorders (FASDs). At the most severe end of the spectrum is Fetal Alcohol Syndrome (FAS), characterized by growth retardation, craniofacial dysmorphology, and neurobehavioral deficits. Studies with animal models, including rodents, have elucidated many molecular and cellular mechanisms involved in the pathophysiology of FASDs. Ethanol administration to pregnant rodents has been used to model human exposure during the first and second trimesters of pregnancy. Third trimester ethanol consumption in humans has been modeled using neonatal rodents. However, few rodent studies have characterized the effect of ethanol exposure during the equivalent to all three trimesters of human pregnancy, a pattern of exposure that is common in pregnant women. Here, we show how to build vapor chambers from readily obtainable materials that can each accommodate up to six standard mouse cages. We describe a vapor chamber paradigm that can be used to model exposure to ethanol, with minimal handling, during all three trimesters. Our studies demonstrate that pregnant dams developed significant metabolic tolerance to ethanol. However, neonatal mice did not develop metabolic tolerance and the number of fetuses, fetus weight, placenta weight, number of pups/litter, number of dead pups/litter, and pup weight were not significantly affected by ethanol exposure. An important advantage of this paradigm is its applicability to studies with genetically-modified mice. Additionally, this paradigm minimizes handling of animals, a major confound in fetal alcohol research.

We demonstrate the construction of alcohol vapor chambers using readily available materials that simultaneously house 6 mouse cages. We further describe their use in a mouse model of fetal alcohol exposure equivalent to all 3&#160;trimesters of human pregnancy. This paradigm exposes animals during gestation and postnatal days 1-12.

Costruzione di vapore Chambers utilizzato per esporre topi da alcol Durante l&#39;equivalente di tutti i tre trimestri di Sviluppo Umano

Exposure to alcohol during development can result in a constellation of morphological and behavioral abnormalities that are collectively known as Fetal ...

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. Despite the discovery of key regulators of uterine contractility, this process is still not fully understood. Transgenic mice provide an ideal model in which to study parturition. Previously, the only method to study uterine contractility in the mouse was ex vivo isometric tension recordings, which are suboptimal for several reasons. The uterus must be removed from its physiological environment, a limited time course of investigation is possible, and the mice must be sacrificed. The recent development of radiometric telemetry has allowed for longitudinal, real-time measurements of in vivo intrauterine pressure in mice. Here, the implantation of an intrauterine telemeter to measure pressure changes in the mouse uterus from mid-pregnancy until delivery is described. By comparing differences in pressures between wild type and transgenic mice, the physiological impact of a gene of interest can be elucidated. This technique should expedite the development of therapeutics used to treat myometrial disorders during pregnancy, including preterm labor.

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

Telemetria intrauterina misurare mouse contrattile Pressione<em&gt; In Vivo</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor initiation, metastasis and resistance to radio- and chemotherapy. Here we describe a specific methodology for culturing primary human pancreatic CSCs as tumor spheres in anchorage-independent conditions. Cells are grown in serum-free, non-adherent conditions in order to enrich for CSCs while their more differentiated progenies do not survive and proliferate during the initial phase following seeding of single cells. This assay can be used to estimate the percentage of CSCs present in a population of tumor cells. Both size (which can range from 35 to 250 micrometers) and number of tumor spheres formed represents CSC activity harbored in either bulk populations of cultured cancer cells or freshly harvested and digested tumors 1,2. Using this assay, we recently found that metformin selectively ablates pancreatic CSCs; a finding that was subsequently further corroborated by demonstrating diminished expression of pluripotency-associated genes/surface markers and reduced in vivo tumorigenicity of metformin-treated cells. As the final step for preclinical development we treated mice bearing established tumors with metformin and found significantly prolonged survival. Clinical studies testing the use of metformin in patients with PDAC are currently underway (e.g., NCT01210911, NCT01167738, and NCT01488552). Mechanistically, we found that metformin induces a fatal energy crisis in CSCs by enhancing reactive oxygen species (ROS) production and reducing mitochondrial transmembrane potential. In contrast, non-CSCs were not eliminated by metformin treatment, but rather underwent reversible cell cycle arrest. Therefore, our study serves as a successful example for the potential of in vitro sphere formation as a screening tool to identify compounds that potentially target CSCs, but this technique will require further in vitro and in vivo validation to eliminate false discoveries.

Pancreatic cancer stem cells (CSCs) can be expanded in vitro using the anchorage-independent sphere culture technique, which represents a powerful tool to study CSC biology and can serve as the first step to develop novel CSC-targeting therapies. Here the methodology for expanding, analyzing and targeting of pancreatic CSCs is provided.

Studiare cancro del pancreas staminali caratteristiche cellulari per lo sviluppo di nuove strategie di trattamento

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is important to standardize protocols to ensure proper analyses and interpretations. In our previous studies, we have measured boldness by monitoring a series of behaviors (time near surface, latency in transitions, number of transitions, and darts) in a 1.5 L trapezoidal tank. Likewise, we quantified aggression by observing bites, lateral displays, darts, and time near an inclined mirror in a rectangular 19 L tank. By dividing a 76 L tank into thirds, we also examined shoaling preferences. The shoaling assay is a highly customizable assay and can be tailored for specific hypotheses. However, protocols for this assay also must be standardized, yet flexible enough for customization. In previous studies, end chambers were either empty, contained 5 or 10 zebrafish, or 5 pearl danios (D. albolineatus). In the following manuscript, we present a detailed protocol and representative data that accompany successful applications of the protocol, which will allow for replication of behavioral syndrome experiments.

This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

L&#39;audacia, l&#39;aggressività e saggi shoaling per Zebrafish comportamentali Sindromi

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

Aprire tracheostomia gastrico acido aspirazione Modello murino di lesioni polmonari acute Risultati in massima acuta non letale Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...

Conducting Maximal and Submaximal Endurance Exercise Testing to Measure Physiological and Biological Responses to Acute Exercise in Humans

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic responses to acute exercise are predominantly influenced by the duration and intensity of the exercise regimen. As exercise is increasingly thought of as a therapeutic treatment and/or diagnostic tool, it is important that standardizable methodologies be utilized to understand the variability and to increase the reproducibility of exercise outputs and measurements of responses to such regimens. To that end, we describe two different cycling exercise regimens that yield different physiologic outputs. In a maximal exercise test, exercise intensity is continually increased with a greater workload resulting in an increasing cardiopulmonary and metabolic response (heart rate, stroke volume, ventilation, oxygen consumption and carbon dioxide production). In contrast, during endurance exercise tests, the demand is increased from that at rest, but is raised to a fixed submaximal exercise intensity resulting in a cardiopulmonary and metabolic response that typically plateaus. Along with the protocols, we provide suggestions on measuring physiologic outputs that include, but are not limited to, heart rate, slow and forced vital capacity, gas exchange metrics, and blood pressure to enable the comparison of exercise outputs between studies. Biospecimens can then be sampled to assess cellular, protein, and/or gene expression responses. Overall, this approach can be easily adapted into both short- and long-term effects of two distinct exercise regimens.

To assess the influence of exercise intensity on physiologic and biologic responses, two different exercise testing protocols were utilized. Methods outlining exercise testing on a cycle ergometer as an incremental maximal oxygen consumption test and endurance, steady state submaximal endurance test are described.

Lo svolgimento di prova per misurare le risposte fisiologiche e biologiche all'esercizio acuto negli esseri umani di esercizio submassimale e massima resistenza

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic ...

Precision Implementation of Minimal Erythema Dose (MED) Testing to Assess Individual Variation in Human Inflammatory Response

Minimal erythema dose (MED) testing is frequently used in clinical settings for determining the smallest amount of ultraviolet (UV) irradiation necessary to produce erythema (inflammatory reddening) on the surface of the skin. In this context, the MED is regarded as a key factor in determining starting doses for UV phototherapy for common skin conditions such as psoriasis and eczema. In research settings, MED testing also has potential to be a powerful tool for assessing within- and between-persons variation in inflammatory responses. However, MED testing has not been widely adopted for use in research settings, likely owing to a lack of published guidelines, which is a barrier to obtaining reproducible results from this assay. Also, protocols and equipment for establishing MED vary widely, making it difficult to compare results across laboratories. Here, we describe a precise and reproducible method to induce and measure superficial erythema using newly designed protocols and methods that can easily be adapted to other equipment and laboratory environments. The method described here includes detail on procedures that will allow extrapolation of a standardized dosage schedule to other equipment so that this protocol can be adapted to any UV radiation source.

Precision Implementation of Minimal Erythema Dose MED Testing to Assess Individual Variation in Human Inflammatory Response

Minimal erythema dose (MED) testing is used to establish dosage schedules for ultraviolet radiation phototherapy. It can assess individual variation in inflammatory response but lacks methodology for achieving reproducible results. Here, we present a precision implementation of MED and demonstrate its ability to capture individual variation in inflammatory response.

Implementazione di precisione dei test di dose minima di erithema (MED) per valutare la variazione individuale nella risposta infiammatoria umana

Minimal erythema dose (MED) testing is frequently used in clinical settings for determining the smallest amount of ultraviolet (UV) irradiation necessary ...

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic fatty liver disease, metabolic liver disease, and liver cancer) requires experimental manipulation of animal models and in vitro cell cultures. Both techniques have limitations, such as the requirement of large numbers of animals for in vivo manipulation. However, in vitro cell cultures do not reproduce the structure and function of the multicellular hepatic environment. The use of precision-cut liver slices is a technique in which uniform slices of viable mouse liver are maintained in laboratory tissue culture for experimental manipulation. This technique occupies an experimental niche that exists between animal studies and in vitro cell culture methods. The presented protocol describes a straightforward and reliable method to isolate and culture precision-cut liver slices from mice. As an application of this technique, ex vivo liver slices are treated with bile acids to simulate cholestatic liver injury and ultimately assess the mechanisms of hepatic fibrogenesis.

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.

Murine Precision-Cut Liver Slices come un modello Ex Vivo di biologia del fegato

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...

Intraoperative Assessment of Resection Margins in Oral Cavity Cancer: This is the Way

The goal of head and neck oncological surgery is complete tumor resection with adequate resection margins while preserving acceptable function and appearance. For oral cavity squamous cell carcinoma (OCSCC), different studies showed that only 15%-26% of all resections are adequate. A major reason for the low number of adequate resections is the lack of information during surgery; the margin status is only available after the final histopathologic assessment, days after surgery.
The surgeons and pathologists at the Erasmus MC University Medical Center in Rotterdam started the implementation of specimen-driven intraoperative assessment of resection margins (IOARM) in 2013, which became the standard of care in 2015. This method enables the surgeon to turn an inadequate resection into an adequate resection by performing an additional resection during the initial surgery. Intraoperative assessment is supported by a relocation method procedure that allows accurate identification of inadequate margins (found on the specimen) in the wound bed.
The implementation of this protocol resulted in an improvement of adequate resections from 15%-40%. However, the specimen-driven IOARM is not widely adopted because grossing fresh tissue is counter-intuitive for pathologists. The fear exists that grossing fresh tissue will deteriorate the anatomical orientation, shape, and size of the specimen and therefore will affect the final histopathologic assessment. These possible negative effects are countered by the described protocol. Here, the protocol for specimen-driven IOARM is presented in detail, as performed at the institute.

The goal of this protocol is to provide a clear overview of specimen-driven intraoperative assessment of resection margins. It is encouraged to implement this protocol to improve patient care at other institutes.