Gli autori desiderano ringraziare tutti i partecipanti, dentisti, e assistenti per la loro partecipazione a questo studio. Ringraziamo il professor Hirotaka Kuwata e Assistant Professor Hirobumi Morisaki, Dipartimento di Microbiologia Orale e Immunologia, Showa University School of Dentistry, per il loro sostegno.

<html> 1. Preparazione della soluzione PMBPAz (Figura 1) <ol><li> Sintetizzare MPC a seguito di un procedimento industriale riportato in precedenza 15. </li><li> Sintetizzare le fotoreattivo MPC polimero PMBPAz usando convenzionale polimerizzazione radicalica di MPC, BMA, e MPAz in etanolo, come descritto in precedenza 14. NOTA: PMBPAz dovrebbe contenere 60%, 30% e 10% (moli%) delle unità monomeriche summenzionati, rispettivamente. Refrigerare il PMBPAz sciolto in etanolo per preparare una soluzione allo 0,5% in peso prima dell&#39;uso. </li><li> Prima di trattare la superficie PMMA, lasciare che la soluzione di etanolo PMBPAz riposare per 30 minuti a temperatura ambiente. </li></ol><img alt="Figura 1" src="/files/ftp_upload/54965/54965fig1.jpg" /> Figura 1. Formula chimica strutturale del fotoreattivo MPC polimero PMBPAz. PMBPAz stato costruito utilizzando un&#39;unità MPC, un&#39;unità BMA, ed una unità MPAz./ftp_upload/54965/54965fig1large.jpg &quot;target =&quot; _ blank &quot;&gt; Clicca qui per vedere una versione più grande di questa figura. 2. PMBPAz Trattamento di un completo PMMA protesi (figura 2) <ol><li> Meccanicamente pulire la protesi per 3 minuti con un pennello protesi. </li><li> Chimicamente pulire la protesi per 5 minuti in un bagno sonicatore ultrasuoni con un detergente protesi compreso ipoclorito di sodio 2,0% e alle seguenti condizioni sonicazione: frequenza, 50/60 Hz; il numero di fase, 1φ; e frequenza di oscillazione, 28 kHz. </li><li> Prima di eseguire la modifica della superficie con PMBPAz, lavare la superficie della protesi per immersione in etanolo. Dopo il lavaggio etanolo, lasciare che la superficie della protesi asciutta per sé, come l&#39;etanolo è un composto volatile. </li><li> Applicare 1 ml di soluzione di etanolo PMBPAz direttamente su tutta la superficie della protesi con un pennello applicatore. Successivamente, tenere la protesi a temperatura ambiente e pressione atmosferica per 10 minuti per permettere al solvente di evaporare in un&#39;atmosfera vapori di etanolo. Lasciare asciugare. Ripetere questi passaggi due volte. </li><li> Irradiare la superficie della protesi con la luce UV per 2 min (a 254 nm). </li><li> Immergere la protesi PMBPAz trattati in acqua distillata per 5 minuti prima dell&#39;uso. </li></ol><img alt="figura 2" src="/files/ftp_upload/54965/54965fig2.jpg" /> Figura 2. Realizzazione di una superficie modificata utilizzando il fotoreattivo MPC polimero PMBPAz. La PMBPAz polimero può covalente legarsi a PMMA mediante l&#39;applicazione di irradiazione UV. <a href="http://ecsource.jove.com/files/ftp_upload/54965/54965fig2large.jpg" target="_blank">Clicca qui per vedere una versione più grande di questa figura.</a> 3. Valutazione di protesi placca Deposizione <ol><li> Utilizzare la protesi PMBPAz trattati a casa per 2 settimane. Come controllo, ripetere la stessa procedura, escludendo il trattamento con il PMBPAz. </li><li> Dopo un paziente ha utilizzato la protesi per 2 settimane (usato about da 12 a 14 ore al giorno), sciacquarlo con acqua di rubinetto per rimuovere eventuali particelle alimentari sfusi, e quindi macchiare con una soluzione di metilene blu Divulgante 0,25% per 1 min. </li><li> Pulire eventuali macchie presenti sulla superficie della protesi in acqua distillata utilizzando un sonicatore bagno ad ultrasuoni per 30 sec. </li><li> Catturare immagini a un angolo di 90 ° su un sedile di cera con una fotocamera digitale utilizzando un sistema a luce bianca. Cattura le immagini con la stessa distanza focale oggetto, tempo di esposizione, e l&#39;intensità di illuminazione. catturare immagini da 3 diverse direzioni per ottenere delle mucose, a destra e sinistra immagini superficie lucidata. </li><li> Calcolare la zona macchiata e quantificare utilizzando un software di analisi di immagine per l&#39;intera superficie della protesi 16. <ol><li> Selezionare l&#39;area della protesi con lo strumento &quot;lazo magnetico&quot; dall&#39;immagine catturata protesi. Successivamente, cambiare il canale di colore rosso per evidenziare l&#39;area della placca dall&#39;immagine zona protesi. Selezionare tutte le aree della placca utilizzando lo strumento &quot;bacchetta magica&quot; (tlivello olerance 8) e la &quot;select simile&quot; strumento dalla zona di protesi. </li><li> Contare il numero totale di pixel per ciascuna immagine. Successivamente, calcolare l&#39;indice percentuale di targa della superficie coperta sulla protesi dal numero di pixel di queste immagini (totale pixel placca / totale pixel superficie della protesi). Valutare le differenze statistiche tra le aree colorate sulle protesi PMBPAz-trattati e non trattati (ANOVA, p &lt;0,05). </li></ol></li></ol>

<ol>
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Soc. 56, 1601-1607 (2008).</li><li>Ishikawa, A., Yoneyama, T., Hirota, K., Miyake, Y., Miyatake, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Professional+oral+health+care+reduces+the+number+of+oropharyngeal+bacteria.">Professional oral health care reduces the number of oropharyngeal bacteria.</a> J. Dent. Res. 87, 594-598 (2008).</li><li>Hosokawa, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prosthodontic+interventions+for+special-needs+patients.">Prosthodontic interventions for special-needs patients.</a> J Prosthodont Res. 58, 69-70 (2014).</li><li>Parvizi, A., Lindquist, T., Schneider, R., Williamson, D., Boyer, D., Dawson, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+the+dimensional+accuracy+of+injection-molded+denture+base+materials+to+that+of+conventional+pressure-pack+acrylic+resin.">Comparison of the dimensional accuracy of injection-molded denture base materials to that of conventional pressure-pack acrylic resin.</a> J. Prosthodont. 13, 83-89 (2004).</li><li>Ishihara, K., Ueda, T., Nakabayashi, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+phospholipid+polylmers+and+their+properties+as+polymer+hydrogel+membranes.">Preparation of phospholipid polylmers and their properties as polymer hydrogel membranes.</a> Polym. J. 22, 355-360 (1990).</li><li>Ishihara, K., Aragaki, R., Ueda, T., Watenabe, A., Nakabayashi, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduced+thrombogenicity+of+polymers+having+phospholipid+polar+groups.">Reduced thrombogenicity of polymers having phospholipid polar groups.</a> J. Biomed. Mater. Res. 24 (8), 1069-1077 (1990).</li><li>Ishihara, K., Ziats, N. P., Tierney, B. P., Nakabayashi, N., Anderson, 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=Protein+adsorption+from+human+plasma+is+reduced+on+phospholipid+polymers.">Protein adsorption from human plasma is reduced on phospholipid polymers.</a> J. Biomed. Mater. Res. 25 (11), 1397-1407 (1991).</li><li>Ishihara, K., Nomura, H., Mihara, T., Kurita, K., Iwasaki, Y., Nakabayashi, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Why+do+phospholipid+polymers+reduce+protein+adsorption?.">Why do phospholipid polymers reduce protein adsorption?.</a> J. Biomed. Mater. Res. 39 (2), 323-330 (1998).</li><li>Ishihara, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioinspired+phospholipid+polymer+biomaterials+for+making+high+performance+artificial+organs.">Bioinspired phospholipid polymer biomaterials for making high performance artificial organs.</a> Sci. Technol. Adv. Mater. 1 (3), 131-138 (2000).</li><li>Iwasaki, Y., Ishihara, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+membrane-inspired+phospholipid+polymers+for+developing+medical+devices+with+excellent+biointerfaces.">Cell membrane-inspired phospholipid polymers for developing medical devices with excellent biointerfaces.</a> Sci. Technol. Adv. Mater. 13 (6), 064101 (10pp) (2012).</li><li>Zhang, N., Chen, C., Melo, M. A., 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=A+novel+protein-repellent+dental+composite+containing+2-methacryloyloxyethyl+phosphorylcholine.">A novel protein-repellent dental composite containing 2-methacryloyloxyethyl phosphorylcholine.</a> Int J Oral Sci. 7, 103-109 (2015).</li><li>Hirota, K., Yumoto, H., Miyamoto, K., 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=MPC-polymer+reduces+adherence+and+biofilm+formation+by+oral+bacteria.">MPC-polymer reduces adherence and biofilm formation by oral bacteria.</a> J Dent Res. 90, 900-905 (2011).</li><li>Fukazawa, K., Ishihara, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+photoreactive+phospholipid+polymers+for+use+in+versatile+surface+modification+of+various+materials+to+obtain+extreme+wettability.">Synthesis of photoreactive phospholipid polymers for use in versatile surface modification of various materials to obtain extreme wettability.</a> ACS Appl. Mater. Interfaces. 5 (15), 6832-6836 (2013).</li><li>Ishihara, K., Ueda, T., Nakabayashi, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+phospholipid+polymers+and+their+properties+as+polymer+hydrogel+membranes.">Preparation of phospholipid polymers and their properties as polymer hydrogel membranes.</a> Polym. J. 22 (5), 355-360 (1990).</li><li>Coulthwaite, L., Verran, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+in+vivo+denture+plaque+assessment+methods.">Evaluation of in vivo denture plaque assessment methods.</a> Br Dent J. 207 (E12), 282-283 (2009).</li><li>He, D., Susanto, H., Ulbricht, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photo-irradiation+for+preparation,+modification+and+stimulation+of+polymeric+membranes.">Photo-irradiation for preparation, modification and stimulation of polymeric membranes.</a> Prog Polym Sci. 34, 62-98 (2009).</li><li>Nakayama, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surface+macromolecular+architectural+designs+using+photo-graft+copolymerization+based+on+photochemistry+of+benzyl+N,N-diethyldithiocarbamate.">Surface macromolecular architectural designs using photo-graft copolymerization based on photochemistry of benzyl N,N-diethyldithiocarbamate.</a> Macromolecules. 29, 8622-8630 (1996).</li><li>Takahashi, N., Iwasa, F., Inoue, Y., Morisaki, H., Ishihara, K., Baba, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+the+durability+and+antiadhesive+action+of+2-methacryloyloxyethyl+phosphorylcholine+grafting+on+an+acrylic+resin+denture+base.">Evaluation of the durability and antiadhesive action of 2-methacryloyloxyethyl phosphorylcholine grafting on an acrylic resin denture base.</a> J. Prosthet. Dent. 112 (2), 194-203 (2014).</li></ol>

Non abbiamo informativa o sostegno finanziario.

Meccanismo della PMBPAz Coating È difficile da legare polimeri MPC ad una superficie PMMA in modo semplice e stabile. Qui, abbiamo utilizzato un nuovo monomero fotoreattivo recante un gruppo phenylazide, MPAz, che polimerizza con altri monomeri attraverso un procedimento di polimerizzazione radicalica convenzionale, risultando in un polimero con catene laterali phenylazide. I gruppi phenylazide sono decomposti mediante irradiazione UV per formare gruppi nitrene, che sono altamente reattivi gru...

sanitaria orale gioca un ruolo importante nella prevenzione di infezioni opportunistiche e polmonite chimica negli anziani fisicamente vulnerabile. Dell&#39;igiene orale, composto da spazzolatura dopo ogni pasto, protesi di pulizia, e la ricezione di cura professionale della salute orale, svolge un ruolo fondamentale nel ridurre l&#39;incidenza di tali malattie 1-3. In particolare, la scarsa igiene orale è fortemente associato con la polmonite da aspirazione negli anziani fisicamente vulnerabili. Pertanto, maggiore attenzione dovrebbe essere prestata alla cura della salute orale dei pazienti anziani, che hanno limitata capacità di pulire le loro protesi da soli 4. Anche nei paesi sviluppati, l&#39;uso di acrilici, protesi rimovibile a base di resina rimane l&#39;opzione di trattamento più comune per i pazienti anziani edentuli 5. Data la loro elevata capacità di assorbimento d&#39;acqua, la superficie microporosa, e la natura idrofoba, basi protesi in resina realizzati da polimetilmetacrilato (PMMA) si accumulano facilmente placca dentale sulle loro superfici, che si chiama protesi placca. 2-metacriloilossietil fosforilcolina polimeri (MPC) sono ben noti materiali polimerici biomedici 6 che hanno notevole antithrombogenicity e la compatibilità dei tessuti a causa della loro elevata resistenza alla assorbimento di proteine e cellule adesione 7-11. MPC rivestimenti polimerici sono stati applicati in vari dispositivi medici per prevenire l&#39;infezione e 12 sono stati suggeriti per avere il potenziale per prevenire l&#39;accumulo di placca protesi su superfici di protesi PMMA 13. La sfida nell&#39;applicazione clinica di polimeri MPC di PMMA dentiere è affidabile legano loro di PMMA superfici in modo semplice e stabile. In precedenza, i polimeri MPC sono stati segnalati per essere stabilmente legato a superfici PMMA da una tecnica di innesto e di avere la possibilità di inibire l&#39;accumulo di placca. Tuttavia, la tecnica di innesto non è semplice e richiede attrezzature specializzate, WHIch fa la sua applicazione clinica impegnativo. Qui, descriviamo una nuova procedura di rivestimento MPC che utilizza polimeri specificamente fosfolipidi fotoreattivi, un metacrilato derivata fotoreattivo, 2-metacriloilossietil-4-azidobenzoate (MPAz) -synthesized per realizzare una varietà di modifiche superficiali di diversi tipi di materiali. MPAz è stato co-polimerizzati con polimeri MPC e n-butil metacrilato (BMA) per produrre poli (MPC- co -BMA- co -MPAz) (PMBPAz). PMBPAz può covalentemente legarsi alle superfici delle protesi PMMA attraverso l&#39;attivazione dei gruppi azide in MPAz sotto ultravioletta (UV) irradiazione 14. Il procedimento non richiede attrezzature specializzate e può essere completata da clinici entro 2 min. Abbiamo inoltre applicato questa procedura in un ambiente clinico e ha dimostrato la sua utilità clinica e l&#39;efficacia nell&#39;inibire la deposizione di placca sulle protesi rimovibili.

<table border="0" cellpadding="0" cellspacing="0" width="695">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:236px;">Ultrasonic Cleaner</td>
			<td style="width:104px;">Aiwa Medical Engineering</td>
			<td style="width:116px;">AU-12C</td>
			<td style="width:239px;">Clean in distilled water before stain the denture</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">UVP UV Crosslinker DL-1000</td>
			<td>Funakoshi</td>
			<td>95-0174-03</td>
			<td>UV light irradiation for 2min before use</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lightbox S</td>
			<td>Suntec</td>
			<td align="right">6542</td>
			<td>capturing in this box after stain the denture</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Adobe Photoshop CS6 Extended</td>
			<td>Adobe</td>
			<td></td>
			<td>Calucurate and quantify</td>
		</tr>
	</tbody>
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2-methacryloyloxyethyl phosphorylcholine polymer treatment of complete dentures to inhibit denture plaque deposition

Removable dentures made of poly (methyl methacrylate) (PMMA) are prone to bacterial adherence and dental plaque formation, which is called denture plaque. Denture plaque-associated infection is a source of serious dental and medical complications in the elderly. 2-Methacryloyloxyethyl phosphorylcholine (MPC) is a well-known biomedical material that exhibits marked antithrombogenicity and tissue compatibility because of its high resistance to protein adsorption and cell adhesion. Therefore, MPC polymer coatings are suggested to have the potential to inhibit plaque deposition on the surface of PMMA dentures. However, coating MPC polymer on the surface of a PMMA denture is a complex procedure that requires specialized equipment, which is regarded as a major barrier to its clinical application. 
Here, we introduce a new MPC polymer treatment procedure that uses poly (MPC-co-BMA-co-MPAz) (PMBPAz) to prevent denture plaque deposition on removable dentures. This procedure enables the MPC coating of PMMA denture surfaces in a simple and stable manner that is resistant to various chemical and mechanical stresses due to the MPC layer of PMBPAz that is covalently bound to the PMMA surface by ultraviolet light irradiation. In addition, the procedure does not require any specialized equipment and can be completed by clinicians within 2 min. We applied this procedure in a clinical setting and demonstrated its clinical utility and efficacy in inhibiting plaque deposition on removable dentures.

trattamento PMBPAz consente il rivestimento MPC di PMMA superfici attraverso un metodo semplice e breve e mantiene l&#39;effetto di inibizione della placca del polimero MPC. Questa procedura non richiede attrezzature specializzate e può essere eseguita alla poltrona dai clinici entro 2 min. Seguendo passo protocollo 2, il trattamento PMBPAz di protesi complete rimovibili da 11 pazienti edentuli è stata condotta con successo in un ambiente clinico senza problemi sostanziali. Le medie in...

Watch this Scientific Journal Video about 2-Methacryloyloxyethyl Phosphorylcholine Polymer Treatment of Complete Dentures to Inhibit Denture Plaque Deposition at JoVE.com

2-Methacryloyloxyethyl Phosphorylcholine Polymer Treatment of Complete Dentures to Inhibit Denture Plaque Deposition

Removable poly(methyl methacrylate) (PMMA) dentures are prone to bacterial adherence and plaque formation. Denture plaque-associated infection is a source of serious dental and medical complications in the elderly. This paper introduces a novel protocol to treat PMMA dentures with 2-methacryloyloxyethyl phosphorylcholine polymer, poly(MPC-co-BMA-co-MPAz), to suppress plaque deposition on PMMA dentures.

2-metacriloilossietil fosforilcolina Polymer Trattamento di protesi totale per l&#39;inibizione della protesi Plaque Deposizione

Removable dentures made of poly (methyl methacrylate) (PMMA) are prone to bacterial adherence and dental plaque formation, which is called denture plaque. ...

2-methacryloyloxyethyl-phosphorylcholine-polymer-treatment-complete

Research

JoVE Journal

Medicine

10.3K Views.  Showa University. Removable poly(methyl methacrylate) (PMMA) dentures are prone to bacterial adherence and plaque formation. Denture plaque-associated infection is a source of serious dental and medical complications in the elderly. This paper introduces a novel protocol to treat PMMA dentures with 2-methacryloyloxyethyl phosphorylcholine polymer, poly(MPC-co-BMA-co-MPAz), to suppress plaque deposition on PMMA dentures.

Video: 2-Methacryloyloxyethyl Phosphorylcholine Polymer Treatment of Complete Dentures to Inhibit Denture Plaque Deposition

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading to an accumulation of lipid-laden intra-luminal plaque, smooth muscle cell proliferation and progressive narrowing of the vessel lumen. The formation of such vulnerable plaques prone to rupture underlies the majority of cases of acute myocardial infarction. The complex molecular and cellular inflammatory cascade is orchestrated by the recruitment of T lymphocytes and macrophages and their paracrine effects on endothelial and smooth muscle cells.1
Molecular imaging in atherosclerosis has evolved into an important clinical and research tool that allows in vivo visualization of inflammation and other biological processes. Several recent examples demonstrate the ability to detect high-risk plaques in patients, and assess the effects of pharmacotherapeutics in atherosclerosis.4 While a number of molecular imaging approaches (in particular MRI and PET) can image biological aspects of large vessels such as the carotid arteries, scant options exist for imaging of coronary arteries.2 The advent of high-resolution optical imaging strategies, in particular near-infrared fluorescence (NIRF), coupled with activatable fluorescent probes, have enhanced sensitivity and led to the development of new intravascular strategies to improve biological imaging of human coronary atherosclerosis.
Near infrared fluorescence (NIRF) molecular imaging utilizes excitation light with a defined band width (650-900 nm) as a source of photons that, when delivered to an optical contrast agent or fluorescent probe, emits fluorescence in the NIR window that can be detected using an appropriate emission filter and a high sensitivity charge-coupled camera. As opposed to visible light, NIR light penetrates deeply into tissue, is markedly less attenuated by endogenous photon absorbers such as hemoglobin, lipid and water, and enables high target-to-background ratios due to reduced autofluorescence in the NIR window. Imaging within the NIR 'window' can substantially improve the potential for in vivo imaging.2,5
Inflammatory cysteine proteases have been well studied using activatable NIRF probes10, and play important roles in atherogenesis. Via degradation of the extracellular matrix, cysteine proteases contribute importantly to the progression and complications of atherosclerosis8. In particular, the cysteine protease, cathepsin B, is highly expressed and colocalizes with macrophages in experimental murine, rabbit, and human atheromata.3,6,7 In addition, cathepsin B activity in plaques can be sensed in vivo utilizing a previously described 1-D intravascular near-infrared fluorescence technology6, in conjunction with an injectable nanosensor agent that consists of a poly-lysine polymer backbone derivatized with multiple NIR fluorochromes (VM110/Prosense750, ex/em 750/780nm, VisEn Medical, Woburn, MA) that results in strong intramolecular quenching at baseline.10 Following targeted enzymatic cleavage by cysteine proteases such as cathepsin B (known to colocalize with plaque macrophages), the fluorochromes separate, resulting in substantial amplification of the NIRF signal. Intravascular detection of NIR fluorescence signal by the utilized novel 2D intravascular NIRF catheter now enables high-resolution, geometrically accurate in vivo detection of cathepsin B activity in inflamed plaque.
In vivo molecular imaging of atherosclerosis using catheter-based 2D NIRF imaging, as opposed to a prior 1-D spectroscopic approach,6 is a novel and promising tool that utilizes augmented protease activity in macrophage-rich plaque to detect vascular inflammation.11,12 The following research protocol describes the use of an intravascular 2-dimensional NIRF catheter to image and characterize plaque structure utilizing key aspects of plaque biology. It is a translatable platform that when integrated with existing clinical imaging technologies including angiography and intravascular ultrasound (IVUS), offers a unique and novel integrated multimodal molecular imaging technique that distinguishes inflammatory atheromata, and allows detection of intravascular NIRF signals in human-sized coronary arteries.

In vivo Near Infrared Fluorescence NIRF Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

We detail a new near-infrared fluorescence (NIRF) catheter for 2-dimensional intravascular molecular imaging of plaque biology in vivo. The NIRF catheter can visualize key biological processes such as inflammation by reporting on the presence of plaque-avid activatable and targeted NIR fluorochromes. The catheter utilizes clinical engineering and power requirements and is targeted for application in human coronary arteries. The following research study describes a multimodal imaging strategy that utilizes a novel in vivo intravascular NIRF catheter to image and quantify inflammatory plaque in proteolytically active inflamed rabbit atheromata.

In vivo Near Infrared fluorescenza (NIRF) intravascolare Imaging Molecolare della placca infiammatoria, un approccio multimodale per Imaging di aterosclerosi

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading ...

Ricerca

Medicina

Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE). In the United States (U.S.), the high morbidity and mortality rates make VTE a serious health concern 1-2. After heart disease and stroke, VTE is the third most common vascular disease 3. In the U.S. alone, there is an estimated 900,000 people affected each year, with 300,000 deaths occurring annually 3. A reliable in vivo animal model to study the mechanisms of this disease is necessary.
The advantages of using the mouse complete stasis model of inferior vena cava thrombosis are several. The mouse model allows for the administration of very small volumes of limited availability test agents, reducing costs dramatically. Most promising is the potential for mice with gene knockouts that allow specific inflammatory and coagulation factor functions to be delineated. Current molecular assays allow for the quantitation of vein wall, thrombus, whole blood, and plasma for assays. However, a major concern involving this model is the operative size constraints and the friability of the vessels. Also, due to the small IVC sample weight (mean 0.005 grams) it is necessary to increase animal numbers for accurate statistical analysis for tissue, thrombus, and blood assays such as real-time polymerase chain reaction (RT-PCR), western blot, enzyme-linked immunosorbent (ELISA), zymography, vein wall and thrombus cellular analysis, and whole blood and plasma assays 4-8.
The major disadvantage with the stasis model is that the lack of blood flow inhibits the maximal effect of administered systemic therapeutic agents on the thrombus and vein wall.

The mouse complete stasis model of inferior vena cava thrombosis yields quantifiable amounts of vein wall tissue and thrombus. It has proven useful for evaluating interactions between the vein wall and the occlusive thrombus and in assessing the progression from acute to chronic inflammation.

Modello del mouse completa stasi della vena cava inferiore Trombosi

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE).  In the United States (U.S.), the high morbidity and ...

The Measurement and Treatment of Suppression in Amblyopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current estimates put the prevalence of amblyopia at approximately 1-3%1-3, the majority of cases being monocular2. Amblyopia is most frequently caused by ocular misalignment (strabismus), blur induced by unequal refractive error (anisometropia), and in some cases by form deprivation.
Although amblyopia is initially caused by abnormal visual input in infancy, once established, the visual deficit often remains when normal visual input has been restored using surgery and/or refractive correction. This is because amblyopia is the result of abnormal visual cortex development rather than a problem with the amblyopic eye itself4,5 . Amblyopia is characterized by both monocular and binocular deficits6,7 which include impaired visual acuity and poor or absent stereopsis respectively. The visual dysfunction in amblyopia is often associated with a strong suppression of the inputs from the amblyopic eye under binocular viewing conditions8. Recent work has indicated that suppression may play a central role in both the monocular and binocular deficits associated with amblyopia9,10 .
Current clinical tests for suppression tend to verify the presence or absence of suppression rather than giving a quantitative measurement of the degree of suppression. Here we describe a technique for measuring amblyopic suppression with a compact, portable device11,12 . The device consists of a laptop computer connected to a pair of virtual reality goggles. The novelty of the technique lies in the way we present visual stimuli to measure suppression. Stimuli are shown to the amblyopic eye at high contrast while the contrast of the stimuli shown to the non-amblyopic eye are varied. Patients perform a simple signal/noise task that allows for a precise measurement of the strength of excitatory binocular interactions. The contrast offset at which neither eye has a performance advantage is a measure of the "balance point" and is a direct measure of suppression. This technique has been validated psychophysically both in control13,14 and patient6,9,11 populations.
In addition to measuring suppression this technique also forms the basis of a novel form of treatment to decrease suppression over time and improve binocular and often monocular function in adult patients with amblyopia12,15,16 . This new treatment approach can be deployed either on the goggle system described above or on a specially modified iPod touch device15.

Amblyopia is a developmental disorder of the visual cortex that is often accompanied by strong suppression of one eye. We present a new technique for measuring and treating interocular suppression in patients with amblyopia that can be deployed using virtual reality goggles or a portable iPod Touch device.

La misurazione e trattamento di soppressione in Ambliopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

The Goeckerman Regimen for the Treatment of Moderate to Severe Psoriasis

Psoriasis is a chronic, immune-mediated inflammatory skin disease affecting approximately 2-3% of the population. The Goeckerman regimen consists of exposure to ultraviolet B (UVB) light and application of crude coal tar (CCT). Goeckerman therapy is extremely effective and relatively safe for the treatment of psoriasis and for improving a patient's quality of life. In the following article, we present our protocol for the Goeckerman therapy that is utilized specifically at the University of California, San Francisco. This protocol details the preparation of supplies, administration of phototherapy and application of topical tar. This protocol also describes how to assess the patient daily, monitor for adverse effects (including pruritus and burning), and adjust the treatment based on the patient's response. Though it is one of the oldest therapies available for psoriasis, there is an absence of any published videos demonstrating the process in detail. The video is beneficial for healthcare providers who want to administer the therapy, for trainees who want to learn more about the process, and for prospective patients who want to undergo treatment for their cutaneous disease.

Psoriasis is a chronic, immune-mediated inflammatory skin disease. The Goeckerman regimen, formulated for the treatment of psoriasis, consists of exposure to ultraviolet B (UVB) light and application of crude coal tar (CCT). The following protocol is for the administration of Goeckerman therapy for the treatment of moderate-to-severe psoriasis.

Il Regimen Goeckerman per il trattamento di moderata a grave psoriasi

Psoriasis is a  chronic, immune-mediated inflammatory skin disease affecting approximately 2-3%  of the population. The Goeckerman regimen consists of ...

A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

Atherosclerosis is a chronic inflammatory disease of the vasculature. There are various methods to study the inflammatory compound in atherosclerotic lesions. Mouse models are an important tool to investigate inflammatory processes in atherogenesis, but these models suffer from the phenotypic and functional differences between the murine and human immune system. In vitro cell experiments are used to specifically evaluate cell type-dependent changes caused by a substance of interest, but culture-dependent variations and the inability to analyze the influence of specific molecules in the context of the inflammatory compound in atherosclerotic lesions limit the impact of the results. In addition, measuring levels of a molecule of interest in human blood helps to further investigate its clinical relevance, but this represents systemic and not local inflammation. Therefore, we here describe a plaque culture model to study human atherosclerotic lesion biology ex vivo. In short, fresh plaques are obtained from patients undergoing endarterectomy or coronary artery bypass grafting and stored in RPMI medium on ice until usage. The specimens are cut into small pieces followed by random distribution into a 48-well plate, containing RPMI medium in addition to a substance of interest such as cytokines or chemokines alone or in combination for defined periods of time. After incubation, the plaque pieces can be shock frozen for mRNA isolation, embedded in Paraffin or OCT for immunohistochemistry staining or smashed and lysed for western blotting. Furthermore, cells may be isolated from the plaque for flow cytometry analysis. In addition, supernatants can be collected for protein measurement by ELISA. In conclusion, the presented ex vivo model opens the possibility to further study inflammatory lesional biology, which may result in identification of novel disease mechanisms and therapeutic targets.

A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

Atherosclerosis is a chronic inflammatory process. This manuscript illustrates an easy to use ex vivo model to investigate fresh carotid or coronary artery plaques. The ex vivo model allows for the investigation of potential substances on the inflammatory milieu in human atherosclerotic lesions and results can be analyzed by various methods.

Un umano<em&gt; Ex Vivo</em&gt; Placca aterosclerotica modello per studiare Lesione Biologia

Atherosclerosis is a chronic inflammatory disease of the vasculature. There are various methods to study the inflammatory compound in atherosclerotic ...

Network Analysis of Foramen Ovale Electrode Recordings in Drug-resistant Temporal Lobe Epilepsy Patients

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control seizures. However, a significant minority of patients continues to exhibit post-operative seizures, even in those cases in which the suspected source of seizures has been correctly localized and resected. The protocol presented here combines a clinical procedure routinely employed during the pre-operative evaluation of temporal lobe epilepsy (TLE) patients with a novel technique for network analysis. The method allows for the evaluation of the temporal evolution of mesial network parameters. The bilateral insertion of foramen ovale electrodes (FOE) into the ambient cistern simultaneously records electrocortical activity at several mesial areas in the temporal lobe. Furthermore, network methodology applied to the recorded time series tracks the temporal evolution of the mesial networks both interictally and during the seizures. In this way, the presented protocol offers a unique way to visualize and quantify measures that considers the relationships between several mesial areas instead of a single area.

This protocol describes a procedure to track the evolution of mesial network measures in temporal lobe epilepsy (TLE) patients. It is based on the combination of intracranial recordings with a novel numerical technique for data analysis. Specifically, we present a protocol for network analyses of foramen ovale recordings.

Network Analysis di forame ovale elettrodi Recordings in farmaco-resistenti epilessia del lobo temporale pazienti

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control ...

Complete and Partial Aortic Occlusion for the Treatment of Hemorrhagic Shock in Swine

Hemorrhage remains the leading cause of preventable deaths in trauma. Endovascular management of non-compressible torso hemorrhage has been at the forefront of trauma care in recent years. Since complete aortic occlusion presents serious concerns, the concept of partial aortic occlusion has gained a growing attention. Here, we present a large animal model of hemorrhagic shock to investigate the effects of a novel partial aortic balloon occlusion catheter and compare it with a catheter that works on the principles of complete aortic occlusion. Swine are anesthetized and instrumented in order to conduct controlled fixed-volume hemorrhage, and hemodynamic and physiological parameters are monitored. Following hemorrhage, aortic balloon occlusion catheters are inserted and inflated in the supraceliac aorta for 60 min, during which the animals receive whole-blood resuscitation as 20% of the total blood volume (TBV). Following balloon deflation, the animals are monitored in a critical care setting for 4 h, during which they receive fluid resuscitation and vasopressors as needed. The partial aortic balloon occlusion demonstrated improved distal mean arterial pressures (MAPs) during the balloon inflation, decreased markers of ischemia, and decreased fluid resuscitation and vasopressor use. As swine physiology and homeostatic responses following hemorrhage have been well-documented and are like those in humans, a swine hemorrhagic shock model can be used to test various treatment strategies. In addition to treating hemorrhage, aortic balloon occlusion catheters have become popular for their role in cardiac arrest, cardiac and vascular surgery, and other high-risk elective surgical procedures.

Here, we present a protocol demonstrating a hemorrhagic shock model in swine that uses aortic occlusion as a bridge to definitive care in trauma. This model has application in testing a wide range of surgical and pharmacological therapeutic strategies.

Completa e parziale occlusione aortica per il trattamento dello Shock emorragico nei suini

Hemorrhage remains the leading cause of preventable deaths in trauma. Endovascular management of non-compressible torso hemorrhage has been at the ...

Predicting Treatment Response to Image-Guided Therapies Using Machine Learning: An Example for Trans-Arterial Treatment of Hepatocellular Carcinoma

Intra-arterial therapies are the standard of care for patients with hepatocellular carcinoma who cannot undergo surgical resection. The objective of this study was to develop a method to predict response to intra-arterial treatment prior to intervention.
The method provides a general framework for predicting outcomes prior to intra-arterial therapy. It involves pooling clinical, demographic and imaging data across a cohort of patients and using these data to train a machine learning model. The trained model is applied to new patients in order to predict their likelihood of response to intra-arterial therapy.
The method entails the acquisition and parsing of clinical, demographic and imaging data from N patients who have already undergone trans-arterial therapies. These data are parsed into discrete features (age, sex, cirrhosis, degree of tumor enhancement, etc.) and binarized into true/false values (e.g., age over 60, male gender, tumor enhancement beyond a set threshold, etc.). Low-variance features and features with low univariate associations with the outcome are removed. Each treated patient is labeled according to whether they responded or did not respond to treatment. Each training patient is thus represented by a set of binary features and an outcome label. Machine learning models are trained using N - 1 patients with testing on the left-out patient. This process is repeated for each of the N patients. The N models are averaged to arrive at a final model.
The technique is extensible and enables inclusion of additional features in the future. It is also a generalizable process that may be applied to clinical research questions outside of interventional radiology. The main limitation is the need to derive features manually from each patient. A popular modern form of machine learning called deep learning does not suffer from this limitation, but requires larger datasets.

Intra-arterial therapies are the standard of care for patients with hepatocellular carcinoma who cannot undergo surgical resection. A method for predicting response to these therapies is proposed. The technique uses pre-procedural clinical, demographic, and imaging information to train machine learning models capable of predicting response prior to treatment.

Predire la risposta di trattamento alle terapie immagine-guida utilizzando Machine Learning: un esempio per Trans-arteriosa trattamento dell'epatocarcinoma

Intra-arterial therapies are the standard of care for patients with hepatocellular carcinoma who cannot undergo surgical resection. The objective of this ...

Direct Intrabronchial Administration to Improve the Selective Agent Deposition Within the Mouse Lung

Intratracheal (IT) administration of experimental agents is an essential technique in murine models of diffuse lung diseases, such as bleomycin-induced pulmonary fibrosis.&#160; However, distribution of intratracheally-administered agents to the distal mouse lung is often asymmetric, with lung parenchymal concentrations increased in the smaller (but equally accessible) left lung of the mouse.&#160; Described in this report is a novel intrabronchial (IB) approach to cannulate the left and/or right lungs of living mice non-operatively.&#160; It is also demonstrated how this approach can be used to selectively administer agents to one lung or adapted (via dose-adjusted IB delivery) to improve the left-right symmetry of lung delivery of experimental agents, thereby improving models of diffuse lung disease such as bleomycin-induced pulmonary fibrosis.

Intratracheal (IT) administration of experimental agents in mice often results in asymmetric delivery to the distal lungs.&#160; In this report, we describe a direct intrabronchial (IB) approach to cannulate each lung in living mice non-operatively.&#160; This approach can be used to selectively administer agents to one lung or may be adapted to improve the symmetric agent delivery to both lungs.

Amministrazione intrabronchiale diretta per migliorare la deposizione selettiva dell'agente all'interno del polmone del mouse

Intratracheal (IT) administration of experimental agents is an essential technique in murine models of diffuse lung diseases, such as bleomycin-induced ...

Nerve Ultrasound Protocol to Detect Dysimmune Neuropathies

Nerve ultrasound is increasingly used in the differential diagnosis of polyneuropathy as a complementary tool to nerve conduction studies. Morphological alterations of the peripheral nerves, such as increasing the cross-sectional area (CSA), have been described in various immune-mediated polyneuropathies. The most prominent morphological changes in nerve ultrasound have been described for the chronic inflammatory demyelinating polyneuropathy (CIDP)-spectrum disease. CIDP may be distinguished from hereditary and other polyneuropathies by measuring the extent and pattern of nerve swellings (CSA increase). Typical findings in demyelinating inflammatory neuropathies are multifocal nerve swellings with inhomogeneous fascicular structure, while CSA increase in demyelinating hereditary neuropathies occurs in a more generalized and homogenous manner. In other non-inflammatory axonal neuropathies, nerves can appear with normal or slight CSA increases, especially in typical entrapment sites. This article presents technical requirements for nerve ultrasound, an examination procedure using a standardized examination protocol, current reference values for the CSA, and typical sonographic pathological findings in patients with inflammatory neuropathies.

This article presents a protocol for nerve ultrasound in polyneuropathies to aid the diagnosis of inflammatory neuropathies.

Protocollo ad ultrasuoni nervosi per rilevare neuropatie disimmuni

Nerve ultrasound is increasingly used in the differential diagnosis of polyneuropathy as a complementary tool to nerve conduction studies. Morphological ...