Vogliamo riconoscere un contratto (WS1763368) da Pfizer Pharmaceuticals per sostenere in questo lavoro. Vorremmo anche ringraziare il saudita Ministero dell&#39;Istruzione Superiore (Mohe) per il loro supporto.

1. L&#39;esercizio <ol><li> Il muscolo di interesse per questo esperimento è stato flessori del gomito (bicipite brachiale). </li><li> La forza muscolare è stata misurata per ogni partecipante di essere in grado di dare ad ogni individuo una resistenza sufficiente. Questa è stata determinata analizzando ogni partecipante per la loro massima resistenza (RM). </li><li> Per testare la RM, abbiamo utilizzato un dispositivo di estensimetro interfacciato con un computer attraverso un (DA-100C) modulo amplificatore Biopac bioelettrica (BIOPAC Systems, Goleta, CA) per misurare la forza muscolare. Il modulo è stato collegato ad un convertitore analogico-100 MP al campionamento convertitore digitale ad una frequenza di 1.000 hertz al secondo, e con una risoluzione di 24 bit (Fig. 10). </li><li> Il dispositivo estensimetro è stata fissata ad un banco con un angolo di 45 °. I soggetti sono stati istruiti a sedersi dietro il dispositivo e resto i gomiti sulla zona imbottita, in modo che la forza sforzo è attraverso i loro polsi. Questo era il modo migliore per assicurare che il soggetto non saràt reclutare qualsiasi altro muscolo che il bicipite (Fig. 11). </li><li> Forza è stato determinato in 3 occasioni, ad ogni contrazione è di 3 secondi di durata con circa 45 secondi che separano le contrazioni. La media delle 3 misurazioni è stata la RM. </li><li> Dopo aver determinato la RM per il muscolo bicipite di ciascun partecipante, la sessione è destinato operazione è stata effettuata con il 35% del loro RM. </li><li> Tutti i soggetti sono stati sottoposti lo stesso esercizio con opportuni pesi ponderati per indurre il dolore muscolare (DOMS). Questa è stata effettuata facendo 4 serie da 25 ripetizioni di curl di concentrazione bicipiti mentre si è seduti su una sedia, e con i gomiti sostenuti sulle cosce (fig. 12). </li><li> Ogni soggetto è stato dato un periodo di 90 secondi di riposo tra ogni serie. Soggetti o ha fatto l&#39;intera serie di 25 ripetizioni, o sono stati istruiti a fermarsi se non sono riusciti a controllare costantemente il peso durante l&#39;esercizio. </li></ol> 2. Infra-Red CaMera Preparazione e configurazione <ol><li> La stanza dove i raggi infrarossi di imaging si svolge è stato fissato a una temperatura costante di ridurre al minimo eventuali distorsioni esterne da differenze di temperatura ambiente, che potrebbe portare a false letture termica. Ai fini di questo esperimento abbiamo avuto una camera a temperatura controllata che è stata mantenuta a circa 23 ° C (+ / - 0,5 ° C). </li><li> La fotocamera è stato fissato a una distanza di 1 metro di distanza, e ad un angolo perpendicolare alla pelle da misurare (Fig. 9a) *. </li><li> Dopo la distanza richiesta è stata costituita, i soggetti sono stati invitati a fermarsi fino a quando l&#39;immagine è stata presa. Questo non dovrebbe richiedere più di un paio di secondi, ma è molto critica per minimizzare i movimenti per assicurare l&#39;accuratezza delle immagini prese. </li><li> E &#39;preferibile che la camera ha più scuro vernice colorata, invece di colori più chiari, per ridurre al minimo le interferenze infrarossi. </li><li> L&#39;illuminazione è anche critico quando si tratta di immagini infrarosse, Becautilizzare fonte di luce che emettono raggi infrarossi onde come illuminazione fluorescente o tungsteno potrebbe dare falsi valori alti. L&#39;opzione migliore illuminazione sarebbe una sala attrezzata con uniforme luci a LED, le luci a LED quasi non producono alcun infrarossi interferenza (fig. 9b) *. </li></ol> * Una serie di test sono stati fatti presso i nostri laboratori utilizzando la telecamera a infrarossi FLIR 660 (Fig. 8), dove abbiamo confrontato le immagini della pelle da diverse angolazioni (0 (perpendicolare), 15, 30, 45 e 60 gradi), e a distanze diverse (1, 2 e 5 metri), dalla pelle, per individuare con precisione la temperatura della pelle. Tutte le immagini sono state confrontate con termocoppie calibrato e la migliore correlazione tra le immagini e le letture termocoppia è stato ad un angolo perpendicolare e ad una distanza di 1 metro di distanza dalla pelle (r = 0,93). Le diverse angolazioni e distanze causato una perdita pixilation, e diminuita la correlazione globale tra le immagini e le letture della termocoppia. 3. Immagine Acquirement <ol><li> Ai fini di questo esperimento, l&#39;immagine del muscolo esercitato è stata presa prima che l&#39;esercizio, e a 24, e 48 ore dopo l&#39;esercizio. </li><li> Il calore del corpo da fonti diverse il target potrebbe disturbare l&#39;immagine termica e dare false letture. Per questo motivo, nessuno dovrebbe essere accanto o dietro il bersaglio. </li><li> In questa indagine, le immagini sia del braccio allenato e non esercitati sono stati prelevati a fini comparativi. Abbiamo esercitato una delle braccia, come è stato detto in precedenza, e l&#39;altro braccio è stato utilizzato come controllo (Fig. 4 e 5). </li><li> Numeri delle immagini dalla telecamera a infrarossi sono stati registrati immediatamente su un foglio separato, in quanto potrebbe essere difficile identificare quale immagine appartiene a chi. </li></ol> 4. Image Processing &amp; analisi <ol><li> Le immagini acquisite IR sono stati elaborati utilizzando il &quot;ThermoVision ExaminIR&quot; Versione software: 1.10.2. </li><li> Dopo aver selezionato l&#39;immagine desiderata perr analisi, quattro regioni di interesse sono stati individuati l&#39;immagine acquisita del braccio utilizzando le caselle statistiche sull&#39;interfaccia del software (Fig. 6). </li><li> Quando le regioni richiesto in tutto il braccio sono stati individuati, il software mostra le medie e le deviazioni standard delle temperature per ciascuna delle regioni selezionate. Possiamo allora o croce confrontare ogni regione singolarmente o per ottenere una temperatura media di tutto il braccio (Fig. 7). </li></ol> 5. Scala analogica visiva e analisi del sangue <ol><li> Una scala analogica visiva (VAS) è stato utilizzato per valutare dolore soggettivo del braccio. La scala era di 10 cm (100 mm) lunga serie scritta &quot;nessun dolore&quot; ad una estremità, e &quot;molto dolorante&quot; alla fine opposto. Ogni partecipante è stato diretto a lasciare un segno lungo la linea da 10 cm a indicare la loro risposta al dolore. </li><li> La VAS sono stati somministrati ai soggetti prima dell&#39;esercizio, 24 ore dopo l&#39;esercizio, e in 48 ore. </li><li> Sangue periferico è stato colraccolti da soggetti per misurare i livelli di concentrazione di mioglobina nel sangue. </li><li> Il sangue è stato tratto dalla vena soggetti antecubitale prima dell&#39;allenamento, 30 minuti dopo l&#39;esercizio era finita, e dopo 48 ore. </li><li> Il sangue è stato centrifugato a 4000 rpm per 10 minuti per separare il siero dalle cellule. I campioni sono stati poi conservati a -80 ° C fino a quando le analisi di mioglobina è stato fatto. </li><li> Mioglobina è stata misurata utilizzando un Tosoh &quot;AIA-360&quot; Automated Analyzer immuno-enzimatico (Tosoh Corp., Tokyo, Giappone). I kit di analisi mioglobina (Myo 025297, ST AIA-PACK Mioglobina) sono stati utilizzati secondo le istruzioni produce. </li></ol> 6. Rappresentante Risultati I risultati di immagini termiche a infrarossi scattate durante questa indagine sono ben rappresentati nella figura 1. Le immagini prese al 3 periodi di tempo (pre-esercizio, 24 ore post-esercizio, e 48 ore post-esercizio) per le braccia esercitato dei 41 soggetti, ha dimostratoun notevole aumento della temperatura il giorno 2 (24 ore dopo l&#39;esercizio) rispetto al pre-esercizio temperature, e le temperature prese a 48 ore. Come mostrato in figura 1, la temperatura cutanea media era 32,80 + / - 1,03 ° C per il giorno 1 (pre-esercizio), e 33,96 + / - 1,46 ° C per il giorno 2 (24 ore dopo l&#39;esercizio fisico), e 32,82 + / - 1,29 per il giorno 3 (48 ore post-esercizio). Questa differenza di temperatura della pelle dal giorno 1 al giorno 2 è risultata significativa (ANOVA p &lt;0,01). Tuttavia, per il non-esercitato braccio, i cambiamenti tra i 3 periodi di tempo non erano evidenti. La Figura 1 mostra che la temperatura cutanea media era 33,08 + / - 0,83 ° C per il giorno 1 (pre-esercizio), e 32,79 + / - 1,42 ° C per il giorno 2 (24 ore dopo l&#39;esercizio fisico), e 33,17 + / - 0,95 per il giorno 3 (48 ore post-esercizio). Questa differenza di temperatura cutanea oltre i 3 giorni non è risultata significativa (ANOVA p = 0,38). I risultati delle letture dal dolore VAS sono mostrati in figura 2. Come si vede nella figfigura 2, il dolore riferito ha avuto un drammatico aumento nei giorni 2 e 3. Livelli di dolore del muscolo esercitato aumentato da 3,6 + / - 6,1 al giorno 1, al 36,3 + / - 22.8 il giorno 2, e il 37,5 + / - 25,3 il giorno 3. Questo aumento dal giorno 1 è risultata significativa (ANOVA p &lt;0,01). I risultati dei livelli di concentrazione della mioglobina sono mostrati in figura 3. Come si vede in questa figura, non c&#39;era quasi alcun cambiamento tra i 2 concentrazioni di mioglobina il giorno 1 (pre, e 30 minuti dopo l&#39;esercizio fisico). Ma il giorno 3, l&#39;aumento della mioglobina era molto grande. Questo aumento il giorno 3 è stato di circa 147 nanogrammi per millilitro (ng / ml) di sangue rispetto ai primi 2 concentrazioni al giorno 1. Concentrazioni di mioglobina sono stati 30,12 + / - 7,66 ng / mL al basale, 31,66 + / - 11,89 ng / ml 30 minuti dopo l&#39;esercizio fisico, e 178,96 + / - 249,51 ng / ml al giorno 3. Questo aumento il giorno 3 è stato altamente significativo (ANOVA p &lt;0,01). Una analisi di correlazione è stata fatta tra le temperature della pelleottenuto dalle immagini a infrarossi e il dolore VAS livelli. Si è constatato che c&#39;era una notevole correlazione tra le letture VAS il giorno 2, e la misurazione della temperatura cutanea il giorno 2. Questa correlazione è risultata significativa (r = 0,312, p &lt;0,05). Tuttavia, non vi era alcuna correlazione evidente tra le letture VAS e le temperature di pelle al giorno 3. Questa correlazione è stata insignificante (r = 0,047, p = 0,77). <img src="/files/ftp_upload/3551/3551fig1.jpg" alt="Figura 1"/> Figura 1. Un grafico rappresentante le differenze di temperatura della pelle tra le braccia esercitate (Diamonds), e poco esercitato braccia (Piazze) del 41 soggetti nel periodo di tempo di 3 giorni. <img src="/files/ftp_upload/3551/3551fig2.jpg" alt="Figura 2"/> Figura 2. Un grafico rappresentativo delle differenze di dolore muscolare percepita misurata con la VAS nel periodo di tempo 3 giorni per tutti i 41 soggetti. Figura 3. Un grafico rappresentativo delle differenze di concentrazioni di mioglobina per tutti i 41 soggetti sopra i 3 periodi di tempo. <img src="/files/ftp_upload/3551/3551fig4.jpg" alt="Figura 4"/> Figura 4. A), una tipica immagine IR del braccio esercitato un soggetto prima dell&#39;esercizio. B) l&#39;immagine a infrarossi dello stesso braccio soggetti 24 ore dopo l&#39;esercizio. <img src="/files/ftp_upload/3551/3551fig5.jpg" alt="Figura 5"/> Figura 5. A), una tipica immagine IR di un soggetto non-esercitato braccio prima dell&#39;esercizio. B) l&#39;immagine a infrarossi dello stesso braccio soggetti 24 ore dopo l&#39;esercizio. <img src="/files/ftp_upload/3551/3551fig6.jpg" alt="Figura 6"/> Figura 6. Un&#39;illustrazione delle 4 regioni di interesse per l&#39;analisi del thermAl immagine del braccio. <img src="/files/ftp_upload/3551/3551fig7.jpg" alt="Figura 7"/> Figura 7. Interfaccia software per la &quot;ThermoVision ExaminIR&quot; che mostra i 4 scatole di interesse su un&#39;immagine IR di un braccio esercitato. Inoltre mostrati sono le interpretazioni statistiche per ogni casella. <img src="/files/ftp_upload/3551/3551fig8.jpg" alt="Figura 8"/> Figura 8. La telecamera IR termico utilizzato per questa indagine (FLIR 660). <img src="/files/ftp_upload/3551/3551fig9.jpg" alt="Figura 9"/> Figura 9. A) L&#39;installazione della telecamera IR 1 metro di distanza dal braccio soggetti. B) Le luci a LED utilizzati nel laboratorio dove sono state scattate le immagini. <img src="/files/ftp_upload/3551/3551fig10.jpg" alt="Figura 10"/> Figura 10. A) I moduli Biopac utilizzato per misurare la forza muscolare. B) Il dispositivo estensimetro fissato a 45 ° panca inclinata e collegato al sistema Biopac. <img src="/files/ftp_upload/3551/3551fig11.jpg" alt="Figura 11"/> Figura 11. Un soggetto tipico esercitando vigore il dispositivo estensimetro. <img src="/files/ftp_upload/3551/3551fig12.jpg" alt="Figura 12"/> Figura 12. Un soggetto sottoposto il protocollo di esercizio per indurre il dolore muscolare.

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Nessun conflitto di interessi dichiarati.

Lo scopo principale di questa indagine è stato quello di valutare l&#39;utilità delle immagini termiche a infrarossi per individuare e misurare dolore muscolare dopo l&#39;esercizio fisico intenso, ed i nostri risultati suggeriscono che l&#39;imaging IR potrebbe essere una tecnica valida per la rilevazione DOMS, soprattutto entro le prime 24 ore di esercizio. Questo non è sorprendente, come Pennes 18 a condizione di un modello molto dettagliato del flusso di calore dal muscolo per la pelle degli arti. Ques...

<table border="1"><tbody><tr><td> Nome del dispositivo </td><td> Azienda </td><td> Numero di catalogo </td><td> Commenti </td></tr><tr><td> Infra-Red telecamera termica </td><td> FLIR </td><td> FLIR SC660 </td><td></td></tr><tr><td> Termica a infrarossi Software per l&#39;analisi </td><td> ThermoVision ExaminIR </td><td> Il software versione 1.10.2 </td><td></td></tr><tr><td> Amplifier Module bioelettrica </td><td> Biopac </td><td> DA100C </td><td> Il DA100C fornisce le impostazioni guadagno variabile, e riferimenti di tensione regolabile. </td></tr><tr><td> Analogico a digitale del modulo convertitore </td><td> Biopac </td><td> MP100 </td><td></td></tr><tr><td> Automatizzata enzima Analyzer immunoenzimatico </td><td> Tosoh </td><td> AIA -360 </td><td> Questo dispositivo è stato utilizzato per analizzare i campioni di sangue, e ottenere le letture mioglobina. </td></tr>
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the use of thermal infra-red imaging to detect delayed onset muscle soreness

Indolenzimento muscolare a insorgenza ritardata (DOMS), noto anche come il danno muscolare indotta da esercizio (EIMD), è comunemente esperienza in individui che sono stati fisicamente inattivi per periodi di tempo prolungati, e iniziare con un attacco inaspettato di esercizio 1-4, ma può anche atleti che si verificano in esercizio oltre i limiti normali di formazione 5. I sintomi associati a questo doloroso fenomeno può variare da dolorabilità muscolare lieve a grave dolore debilitante 1,3,5. L&#39;intensità di questi sintomi e gli aumenti disagio legate entro le prime 24 ore dopo la cessazione dell&#39;esercizio, e picchi tra 24-72 dopo 1,3 ore di esercizio fisico. Per questo motivo, DOMS è una delle forme più comuni ricorrenti di lesioni sportive che possono influire sulle prestazioni di un individuo, e diventano 1,4 intimidatorio per molti. Negli ultimi 3 decenni, il fenomeno DOMS ha acquisito una notevole quantitàdi interesse tra i ricercatori e specialisti in fisiologia dell&#39;esercizio, dello sport e campi di riabilitazione 6. C&#39;è stata una serie di studi pubblicati indagare questo evento doloroso in relazione ai suoi meccanismi di base, interventi di trattamento e strategie preventive 1-5,7-12. Tuttavia, è evidente dalla letteratura che DOMS non è una patologia facile da quantificare, poiché vi è una quantità gamma di variabilità tra gli strumenti di misura e metodi utilizzati per quantificare questa condizione 6. E &#39;ovvio che nessun accordo è stato fatto su una misura migliore valutazione per DOMS, il che rende difficile verificare se un intervento specifico in realtà aiuta a diminuire i sintomi associati a questo tipo di dolore o meno. Così, DOMS può essere visto come un po &#39;ambigua, perché dipendono da molti studi sulla misurazione dolore utilizzando una scala analogica visiva (VAS) 10,13-15, che è soggettiva piuttosto che una misura oggettiva. Anche se l&#39;agobiopsie del muscolo, e livelli ematici di proteine ​​myofibre potrebbe essere considerato un gold standard per alcuni 6, grandi variazioni in alcune di queste proteine ​​del sangue sono stati documentati 6,16, in aggiunta ai rischi elevati talvolta associata a tecniche invasive. Pertanto, nell&#39;ambito dell&#39;inchiesta in corso, abbiamo testato una termica a infrarossi (IR) tecnica di imaging della pelle sopra il muscolo esercitato per rilevare il dolore muscolare associato. Termografia a infrarossi è stata utilizzata, e trovato per avere successo nel rilevare diversi tipi di malattie e infezioni fin dal 1950 17. Ma sorprendentemente, nei pressi di niente è stato fatto sul DOMS e cambiamenti nella temperatura della pelle. Lo scopo principale di questa indagine è stato quello di esaminare i cambiamenti in DOMS utilizzando questa tecnica sicura e non invasiva.

Watch this Scientific Journal Video about The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness at JoVE.com

The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness

Lo scopo di questa indagine è stato quello di valutare se l&#39;utilizzo di raggi infrarossi della telecamera termica è un valido strumento per rilevare e quantificare il dolore muscolare dopo l&#39;esercizio.

L&#39;uso di Thermal Imaging Infra-Red per rilevare indolenzimento muscolare a insorgenza ritardata

Delayed onset muscle soreness (DOMS), also known as exercise induced muscle damage (EIMD), is commonly experienced in individuals who have been physically ...

the-use-thermal-infra-red-imaging-to-detect-delayed-onset-muscle

Research

JoVE Journal

Medicine

19.1K Views.  Loma Linda University. Lo scopo di questa indagine è stato quello di valutare se l'utilizzo di raggi infrarossi della telecamera termica è un valido strumento per rilevare e quantificare il dolore muscolare dopo l'esercizio.

Video: The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness

Thermal Ablation for the Treatment of Abdominal Tumors

Percutaneous thermal ablation is an emerging treatment option for many tumors of the abdomen not amenable to conventional treatments. During a thermal ablation procedure, a thin applicator is guided into the target tumor under imaging guidance. Energy is then applied to the tissue until temperatures rise to cytotoxic levels (50-60 &deg;C). Various energy sources are available to heat biological tissues, including radiofrequency (RF) electrical current, microwaves, laser light and ultrasonic waves. Of these, RF and microwave ablation are most commonly used worldwide.
During RF ablation, alternating electrical current (~500 kHz) produces resistive heating around the interstitial electrode. Skin surface electrodes (ground pads) are used to complete the electrical circuit. RF ablation has been in use for nearly 20 years, with good results for local tumor control, extended survival and low complication rates1,2. Recent studies suggest RF ablation may be a first-line treatment option for small hepatocellular carcinoma and renal-cell carcinoma3-5. However, RF heating is hampered by local blood flow and high electrical impedance tissues (eg, lung, bone, desiccated or charred tissue)6,7. Microwaves may alleviate some of these problems by producing faster, volumetric heating8-10. To create larger or conformal ablations, multiple microwave antennas can be used simultaneously while RF electrodes require sequential operation, which limits their efficiency. Early experiences with microwave systems suggest efficacy and safety similar to, or better than RF devices11-13.
Alternatively, cryoablation freezes the target tissues to lethal levels (-20 to -40 &deg;C). Percutaneous cryoablation has been shown to be effective against RCC and many metastatic tumors, particularly colorectal cancer, in the liver14-16. Cryoablation may also be associated with less post-procedure pain and faster recovery for some indications17. Cryoablation is often contraindicated for primary liver cancer due to underlying coagulopathy and associated bleeding risks frequently seen in cirrhotic patients. In addition, sudden release of tumor cellular contents when the frozen tissue thaws can lead to a potentially serious condition known as cryoshock 16.
Thermal tumor ablation can be performed at open surgery, laparoscopy or using a percutaneous approach. When performed percutaneously, the ablation procedure relies on imaging for diagnosis, planning, applicator guidance, treatment monitoring and follow-up. Ultrasound is the most popular modality for guidance and treatment monitoring worldwide, but computed tomography (CT) and magnetic resonance imaging (MRI) are commonly used as well. Contrast-enhanced CT or MRI are typically employed for diagnosis and follow-up imaging.

A thermal tumor ablation procedure is described. The entire procedure is detailed, including pretreatment planning and imaging studies, anesthesia, adjuvant techniques to facilitate a percutaneous approach, imaging guidance of the ablation device to the tumor, thermal treatment, post-treatment care and follow-up.

Ablazione termica nel trattamento dei tumori addominali

Percutaneous thermal ablation is an emerging treatment option for many tumors of the abdomen not amenable to conventional treatments. During a thermal ...

Ricerca

Medicina

Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging

In 2010 approximately 68,720 melanomas will be diagnosed in the US alone, with around 8,650 resulting in death 1. To date, the only effective treatment for melanoma remains surgical excision, therefore, the key to extended survival is early detection 2,3. Considering the large numbers of patients diagnosed every year and the limitations in accessing specialized care quickly, the development of objective in vivo diagnostic instruments to aid the diagnosis is essential. New techniques to detect skin cancer, especially non-invasive diagnostic tools, are being explored in numerous laboratories. Along with the surgical methods, techniques such as digital photography, dermoscopy, multispectral imaging systems (MelaFind), laser-based systems (confocal scanning laser microscopy, laser doppler perfusion imaging, optical coherence tomography), ultrasound, magnetic resonance imaging, are being tested. Each technique offers unique advantages and disadvantages, many of which pose a compromise between effectiveness and accuracy versus ease of use and cost considerations. Details about these techniques and comparisons are available in the literature 4.
Infrared (IR) imaging was shown to be a useful method to diagnose the signs of certain diseases by measuring the local skin temperature. There is a large body of evidence showing that disease or deviation from normal functioning are accompanied by changes of the temperature of the body, which again affect the temperature of the skin 5,6. Accurate data about the temperature of the human body and skin can provide a wealth of information on the processes responsible for heat generation and thermoregulation, in particular the deviation from normal conditions, often caused by disease. However, IR imaging has not been widely recognized in medicine due to the premature use of the technology 7,8 several decades ago, when temperature measurement accuracy and the spatial resolution were inadequate and sophisticated image processing tools were unavailable. This situation changed dramatically in the late 1990s-2000s. Advances in IR instrumentation, implementation of digital image processing algorithms and dynamic IR imaging, which enables scientists to analyze not only the spatial, but also the temporal thermal behavior of the skin 9, allowed breakthroughs in the field.
In our research, we explore the feasibility of IR imaging, combined with theoretical and experimental studies, as a cost effective, non-invasive, in vivo optical measurement technique for tumor detection, with emphasis on the screening and early detection of melanoma 10-13. In this study, we show data obtained in a patient study in which patients that possess a pigmented lesion with a clinical indication for biopsy are selected for imaging. We compared the difference in thermal responses between healthy and malignant tissue and compared our data with biopsy results. We concluded that the increased metabolic activity of the melanoma lesion can be detected by dynamic infrared imaging.

We demonstrated that malignant pigmented lesions with increased metabolic activity generate quantifiable amounts of heat and the measurement of the transient thermal response of the skin to a cooling excitation allows quantitative identification of melanoma and other skin cancers (vs. non-proliferative nevi) at an early stage of the disease.

Visualizzazione quantitativa e diagnosi del tumore della pelle utilizzando Dynamic Thermal Imaging

In 2010 approximately 68,720 melanomas will be diagnosed in the US alone, with around 8,650 resulting in death 1. To date, the only effective treatment ...

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

Uso di un modello animale di sepsi per valutare nuove terapie a base di erbe

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel underlying neurobiological mechanisms behind drug action and neurotransmitter-related disorders, such as depression and ADHD. Like most of the imaging methods (PET, SPECT, CT) it represents a progress in the investigation of brain disorders and the related function of neurotransmitter pathways in a non-invasive way with respect of the overall neuronal connectivity. Moreover it also provides the ideal tool for translation to clinical investigations. MRI, while still behind in molecular imaging strategies compared to PET and SPECT, has the great advantage to have a high spatial resolution and no need for the injection of a contrast-agent or radio-labeled molecules, thereby avoiding the repetitive exposure to ionizing radiations. Functional MRI (fMRI) is extensively used in research and clinical setting, where it is generally combined with a psycho-motor task. phMRI is an adaptation of fMRI enabling the investigation of a specific neurotransmitter system, such as serotonin (5-HT), under physiological or pathological conditions following activation via administration of a specific challenging drug.
The aim of the method described here is to assess brain 5-HT function in free-breathing animals. By challenging the 5-HT system while simultaneously acquiring functional MR images over time, the response of the brain to this challenge can be visualized. Several studies in animals have already demonstrated that drug-induced increases in extracellular levels of e.g. 5-HT (releasing agents, selective re-uptake blockers, etc) evoke region-specific changes in blood oxygenation level dependent (BOLD) MRI signals (signal due to a change of the oxygenated/deoxygenated hemoglobin levels occurring during brain activation through an increase of the blood supply to supply the oxygen and glucose to the demanding neurons) providing an index of neurotransmitter function. It has also been shown that these effects can be reversed by treatments that decrease 5-HT availability16,13,18,7. In adult rats, BOLD signal changes following acute SSRI administration have been described in several 5-HT related brain regions, i.e. cortical areas, hippocampus, hypothalamus and thalamus9,16,15. Stimulation of the 5-HT system and its response to this challenge can be thus used as a measure of its function in both animals and humans2,11.

The goal of this technique is to assess serotonin (5-HT) neurotransmitter function in the live and free-breathing animal with pharmacological magnetic resonance imaging (phMRI) and an intravenous challenge with a selective serotonin reuptake inhibitor (SSRI), fluoxetine.

L&#39;uso di Farmacologiche-sfida fMRI in ricerca pre-clinica: Applicazione del sistema 5-HT

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel ...

Molecular Imaging to Target Transplanted Muscle Progenitor Cells

Duchenne muscular dystrophy (DMD) is a severe genetic neuromuscular disorder that affects 1 in 3,500 boys, and is characterized by progressive muscle degeneration1, 2. In patients, the ability of resident muscle satellite cells (SCs) to regenerate damaged myofibers becomes increasingly inefficient4. Therefore, transplantation of muscle progenitor cells (MPCs)/myoblasts from healthy subjects is a promising therapeutic approach to DMD. A major limitation to the use of stem cell therapy, however, is a lack of reliable imaging technologies for long-term monitoring of implanted cells, and for evaluating its effectiveness. Here, we describe a non-invasive, real-time approach to evaluate the success of myoblast transplantation. This method takes advantage of a unified fusion reporter gene composed of genes (firefly luciferase [fluc], monomeric red fluorescent protein [mrfp] and sr39 thymidine kinase [sr39tk]) whose expression can be imaged with different imaging modalities9, 10. A variety of imaging modalities, including positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), optical imaging, and high frequency 3D-ultrasound are now available, each with unique advantages and limitations11. Bioluminescence imaging (BLI) studies, for example, have the advantage of being relatively low cost and high-throughput. It is for this reason that, in this study, we make use of the firefly luciferase (fluc) reporter gene sequence contained within the fusion gene and bioluminescence imaging (BLI) for the short-term localization of viable C2C12 myoblasts following implantation into a mouse model of DMD (muscular dystrophy on the X chromosome [mdx] mouse)12-14. Importantly, BLI provides us with a means to examine the kinetics of labeled MPCs post-implantation, and will be useful to track cells repeatedly over time and following migration. Our reporter gene approach further allows us to merge multiple imaging modalities in a single living subject; given the tomographic nature, fine spatial resolution and ability to scale up to larger animals and humans10,11, PET will form the basis of future work that we suggest may facilitate rapid translation of methods developed in cells to preclinical models and to clinical applications.

A non-invasive means to evaluate the success of myoblast transplantation is described. The method takes advantage of a unified fusion reporter gene composed of genes whose expression can be imaged with different imaging modalities. Here, we make use of a fluc reporter gene sequence to target cells via bioluminescence imaging.

Imaging molecolare a Target trapiantate cellule progenitrici muscolari

Duchenne muscular dystrophy (DMD) is a severe genetic neuromuscular disorder that affects 1 in 3,500 boys, and is characterized by progressive muscle ...

High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum

Over the recent years, antibodies against surface and conformational proteins involved in neurotransmission have been detected in autoimmune CNS diseases in children and adults. These antibodies have been used to guide diagnosis and treatment. Cell-based assays have improved the detection of antibodies in patient serum. They are based on the surface expression of brain antigens on eukaryotic cells, which are then incubated with diluted patient sera followed by fluorochrome-conjugated secondary antibodies. After washing, secondary antibody binding is then analyzed by flow cytometry. Our group has developed a high-throughput flow cytometry live cell-based assay to reliably detect antibodies against specific neurotransmitter receptors. This flow cytometry method is straight forward, quantitative, efficient, and the use of a high-throughput sampler system allows for large patient cohorts to be easily assayed in a short space of time. Additionally, this cell-based assay can be easily adapted to detect antibodies to many different antigenic targets, both from the central nervous system and periphery. Discovering additional novel antibody biomarkers will enable prompt and accurate diagnosis and improve treatment of immune-mediated disorders.

Over the recent years, live cell-based assays have been used successfully to detect antibodies against surface and conformational antigens. Here, we describe a method using high-throughput flow cytometry enabling the analysis of large cohorts of patients. Detection of novel antibodies will improve diagnosis and treatment of immune-mediated disorders.

High-throughput Citometria a Flusso test cellulare per individuare gli anticorpi per N-metil-D-aspartato Receptor o dopamina-2 recettore nel siero umano

Over the recent years, antibodies against surface and conformational proteins involved in neurotransmission have been detected in autoimmune CNS diseases ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

Modello murino per l&#39;imaging non invasivo per rilevare e monitorare Ovarian Cancer Ricorrenza

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

Quantitative Magnetic Resonance Imaging of Skeletal Muscle Disease

Quantitative magnetic resonance imaging (qMRI) describes the development and use of MRI to quantify physical, chemical, and/or biological properties of living systems. Neuromuscular diseases often exhibit a temporally varying, spatially heterogeneous, and multi-faceted pathology. The goal of this protocol is to characterize this pathology using qMRI methods. The MRI acquisition protocol begins with localizer images (used to locate the position of the body and tissue of interest within the MRI system), quality control measurements of relevant magnetic field distributions, and structural imaging for general anatomical characterization. The qMRI portion of the protocol includes measurements of the longitudinal and transverse relaxation time constants (T1 and T2, respectively). Also acquired are diffusion-tensor MRI data, in which water diffusivity is measured and used to infer pathological processes such as edema. Quantitative magnetization transfer imaging is used to characterize the relative tissue content of macromolecular and free water protons. Lastly, fat-water MRI methods are used to characterize fibro-adipose tissue replacement of muscle. In addition to describing the data acquisition and analysis procedures, this paper also discusses the potential problems associated with these methods, the analysis and interpretation of the data, MRI safety, and strategies for artifact reduction and protocol optimization.

Neuromuscular diseases often exhibit a temporally varying, spatially heterogeneous, and multi-faceted pathology. The goal of this protocol is to characterize this pathology using non-invasive magnetic resonance imaging methods.

Quantitative Magnetic Resonance Imaging di malattia muscolare scheletrico

Quantitative magnetic resonance imaging (qMRI) describes the development and use of MRI to quantify physical, chemical, and/or biological properties of ...

Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation

Cardiovascular disease is the leading cause of morbidity and mortality in the world. Atherosclerotic plaques, consisting of lipid-laden macrophages and calcification, develop in the coronary arteries, aortic valve, aorta, and peripheral conduit arteries and are the hallmark of cardiovascular disease. In humans, imaging with computed tomography allows for the quantification of vascular calcification; the presence of vascular calcification is a strong predictor of future cardiovascular events. Development of novel therapies in cardiovascular disease relies critically on improving our understanding of the underlying molecular mechanisms of atherosclerosis. Advancing our knowledge of atherosclerotic mechanisms relies on murine and cell-based models. Here, a method for imaging aortic calcification and macrophage infiltration using two spectrally distinct near-infrared fluorescent imaging probes is detailed. Near-infrared fluorescent imaging allows for the ex vivo quantification of calcification and macrophage accumulation in the entire aorta and can be used to further our understanding of the mechanistic relationship between inflammation and calcification in atherosclerosis. Additionally, a method for isolating and culturing animal aortic vascular smooth muscle cells and a protocol for inducing calcification in cultured smooth muscle cells from either murine aortas or from human coronary arteries is described. This in vitro method of modeling vascular calcification can be used to identify and characterize the signaling pathways likely important for the development of vascular disease, in the hopes of discovering novel targets for therapy.

Vascular calcification is an important predictor of and contributor to human cardiovascular disease. This protocol describes methods for inducing calcification of cultured primary vascular smooth muscle cells and for quantifying calcification and macrophage burden in animal aortas using near-infrared fluorescence imaging.

Calcificazione vascolare cellule muscolari lisce e imaging di aortica calcificazioni e infiammazione

Cardiovascular disease is the leading cause of morbidity and mortality in the world.  Atherosclerotic plaques, consisting of lipid-laden macrophages and ...

In Vivo Evaluation of the Mechanical and Viscoelastic Properties of the Rat Tongue

The tongue is a highly innervated and vascularized muscle hydrostat on the floor of the mouth of most vertebrates. Its primary functions include supporting mastication and deglutition, as well as taste-sensing and phonetics. Accordingly, the strength and volume of the tongue can impact the ability of vertebrates to accomplish basic activities such as feeding, communicating, and breathing. Human patients with sleep apnea have enlarged tongues, characterized by reduced muscle tone and increased intramuscular fat that can be visualized and quantified by magnetic resonance imaging (MRI). The abilities to measure force generation and viscoelastic properties of the tongue constitute important tools for obtaining functional information to correlate with imaging data. Here, we present techniques for measuring tongue force production in anesthetized Zucker rats via electrical stimulation of the hypoglossal nerves and for determining the viscoelastic properties of the tongue by applying passive Lissajous force/deformation curves.

In Vivo Evaluation of the Mechanical and Viscoelastic Properties of the Rat Tongue

We describe a surgical procedure in an anesthetized rat model for determining the muscle tone and viscoelastic properties of the tongue. The procedure involves specific stimulation of the hypoglossal nerves and application of passive Lissajous force/deformation curves to the muscle.