We would like to thank the whole Service of Neuroradiology. We would like to thank Lis Prussen for her work in the library.

NOTA: Questo protocollo segue le linee guida del Centre Hospitalier de Luxembourg in Lussemburgo. 1 Preparazione del tensore di diffusione Imaging per via visiva di Neurochirurgia e Follow Up <ol style="margin-left: 40px;"><li> Eseguire una risonanza magnetica scansione almeno un giorno prima dell&#39;intervento chirurgico rigorosamente assiale con 32 indicazioni di pendenza e un b0-immagine. Tenere in stretto contatto con l&#39;unità di neuroradiologia in qualsiasi momento. NOTA: Rendere chiaro al neuroradiologo che le immagini dopo l&#39;intervento chirurgico sono le stesse di quelle prima dell&#39;operazione. </li><li> Utilizzando un 3-Tesla MRI, eseguire una scansione 3DT1 ponderate e DTI-sequenza. Eseguire una sequenza 3DT1 ponderato dopo l&#39;intervento pure. </li></ol> 2 Utilizzo della stazione di pianificazione <ol><li> Trasferire i dati di scansione T2-pesate, 3DT1 ponderato e DTI-sequenza per imaging digitale e le comunicazioni in medicina (DICOM). Questa procedura richiede fino a 7 min. NOTA: Don `t fermare la procedura di prire dopo aver trasferito tutte le sequenze. È possibile fermare e continuare successivamente a seconda della data di intervento chirurgico. </li><li> Aprire il programma di sistema di navigazione chirurgica. Fare clic su File e poi su Importa DICOM. Ripetere questa procedura per tre volte per tutte le sequenze di cui sopra. <ol><li> Fare clic su Aggiungi per visualizzare. Aggiungere ogni sequenza separatamente. Don `t cercare di procedere con Vista. </li></ol></li><li> Fare clic su Strumenti. Aprire preparazione DTI Tensor. Osservare una nuova finestra al centro dello schermo. </li><li> Completare le seguenti quattro fasi. <ol><li> Eseguire Assegnazione gradiente come il primo passo. <ol><li> Modificare il valore-b da 1.000 a 800 s / mm ² in basso a destra della finestra. </li><li> Regolare la soglia in alto a destra della finestra. Farlo manualmente semplicemente scrivendo un numero o spostamento di un cursore. 20 potrebbe essere un valore accettabile. Si tratta di una esperienza personale e non è obbligatorio. </li></ol></li><li> Eseguire la registrazione Gradiente nel secondo passo. <ol> &lt;li&gt; Fare clic sul pulsante Tutti Auto. Questa procedura richiede fino a 5 min. </li><li> Fare clic su Verifica tutte le registrazioni. Senza verifica delle registrazioni non è possibile continuare. </li></ol></li><li> Eseguire Coregistrazione come il terzo passo. <ol><li> COREGISTER MR1 e MR2 immagini B0 manualmente. Alla fine Verifica tutte le registrazioni. NOTA: E &#39;possibile eseguire questo passaggio automaticamente. Tuttavia, i risultati non sono sempre soddisfacenti, alla fine. </li></ol></li><li> Eseguire Tensor computazione come il quarto e ultimo passo, <ol><li> Assicurarsi che FA / DEC / ADC sono su. Se non fai clic su ON. </li><li> Fare clic su Calcola. Questa procedura richiederà solo alcuni secondi. </li></ol></li></ol></li><li> Salvare tutti i dati e continuare con fibertracking. Non interrompere senza salvare tutto. </li></ol> 3 Fibertracking NOTA: anatomico conoscenza della via visiva è molto importante per il risultato positivo. <ol style = &quot;margin-left: 40px;&quot;&gt; <li> Preparatevi a scoprire i tre punti importanti in cui le fibre devono passare attraverso. </li><li> Determinare il chiasma ottico utilizzando conoscenze anatomiche. <ol><li> Utilizzare un ROI come punto di partenza e lasciare che le fibre passano attraverso. ROI sono definiti dal medico. </li><li> In alternativa, segmento regione sospetto. Clicca Segmentazione in basso a sinistra e apparirà un&#39;altra finestra. Aree segmentate sono anatomicamente aree definite. <ol><li> Dipingi la regione manualmente. Scorrere verso l&#39;alto e verso il basso per includere tutto il chiasma ottico. Salvare la procedura e tornare indietro. </li></ol></li><li> Traccia le fibre sia della regione di interesse o dalla zona segmentati o entrambi. </li><li> Le fibre raggiungono il nucleo genicolato sinistra (LGN), che è il secondo punto importante della via visiva. L&#39;angolo massimo è stato 50 il rischio di falsi tratti salirà con se l&#39;angolo è troppo alto. <ol><li> C&#39;è la possibilità di segmentare il LGN come mostrato con il chiasma otticoe poi seguire le fibre. Dopo aver segmentato il chiasma ottico, fibre pista che corrono dal LGN e finire nel chiasma ottico o viceversa. </li></ol></li><li> Segmento della corteccia visiva. Procedere come nel caso del chiasma ottico. Questo potrebbe richiedere del tempo come immagine 3DT1 ponderate contiene 160 fette. </li><li> Traccia le fibre della corteccia visiva al LGN. È possibile seguire loro dal LGN alla corteccia visiva pure. </li><li> Se la corteccia visiva è invaso da un tumore o edema quindi utilizzare una regione di interesse in luogo di una superficie segmentata e poi lasciare che le fibre correre in direzione del LGN. NOTA: Se l&#39;edema è segmentato a volte potrebbe invadere la corteccia visiva, allora poi la corteccia visiva potrebbe non essere in grado di segmentazione del tutto perché il computer può `t distinguere tra di loro. Quel `s perché è necessario mettere un ROI. </li><li> Ripetere tutto per l&#39;altro emisfero. </li><li> Inizia con l&#39;emisfero sano prima. NOTA: SiÈ possibile iniziare con l&#39;altra troppo ma potrebbe essere più facile per tenere traccia delle fibre dell&#39;emisfero sano prima di diventare una prima idea della situazione. Non è obbligatorio, è solo un consiglio. </li></ol></li><li> Segmento la lesione cerebrale e l&#39;edema. Procedere come indicato al 3.2.2. <ol><li> Assegnare un colore per ogni area segmentato o lesione per distinguere meglio. </li></ol></li><li> Salvare la procedura dopo ogni passaggio in caso di eventi imprevisti o in caso di emergenza. </li><li> Esportare tutti i dati in locale. E &#39;possibile esportarlo in sala operatoria direttamente, ma `t di isn consigliato. <ol><li> Premere File e quindi esportare oggetti 3D. Assicurarsi di esportare solo l&#39;esame di navigazione. </li><li> Don `t esportare l&#39;esame Hybrid. </li></ol></li><li> Inserisci cranica. Scegli il paziente giusto, quindi premere Stealthmerge. Scegliere le immagini 3DT1 ponderato esame di riferimento. </li><li> Creazione di un modello 3D e inserire tutto. </li><li> Importare il i datin la sala operatoria. </li></ol>

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The authors declare that they have no competing financial interests.

DTI è una tecnica che consente di visualizzare il neurochirurgo tratti di sostanza bianca in vivo 8. Il percorso visivo è uno di questi tratti. Anche se questo metodo fornisce ai medici con nuove possibilità per quanto riguarda il trattamento di pazienti con lesioni riguardanti le regioni eloquenti del cervello dobbiamo dire che alcuni limiti di questa tecnica esistono ancora. La prima e più ovvia sfida è spostamento del cervello, che rimane una questione in esame 4. Dopo l&#39;apertu...

Diffusione tensore di imaging (DTI) è utilizzato per ritrarre WMT non invasivo nel cervello umano 1. E &#39;stato utilizzato negli ultimi dieci anni per ridurre il rischio di danneggiare le aree eloquenti del cervello durante l&#39;intervento chirurgico 1. DTI è stata eseguita in una cinquantina di pazienti tra il marzo 2012 e il gennaio 2014 e il ritrarre il percorso visivo. DTI potrebbe migliorare la conservazione delle aree eloquenti del cervello durante l&#39;intervento chirurgico, fornendo importanti informazioni sulla localizzazione anatomica dei tratti di sostanza bianca. E &#39;stato inserito nella pianificazione strategica per la resezione di lesioni cerebrali complesse 1. Tuttavia, la rappresentazione del percorso visivo rimane una sfida, perché non esiste uno standard per i parametri del DTI, la collocazione dei volumi di semi e interpretazione dei risultati 12. Diversi algoritmi sono stati implementati finora 19-21. Alcuni approcci concentrati su metodi deterministici 19, 22-25. Altri usavano metodi probabilistici, 26,27,29. Più di recente, le tecniche che utilizzano campi tensoriali Q-ball, immagini spettrali diffusione e ad alta risoluzione angolare Diffusion Imaging (HARDI) vengono utilizzati per raffigurare tratti di sostanza bianca tra gli altri il percorso visivo 1,13-15,18. Tuttavia, il tempo necessario per HARDI è significativamente più lungo di 45 min, il software non è disponibile in commercio e sottolinea applicazioni scientifiche 18. Il periodo di insegnamento per HARDI sembra essere più lungo per DTI 18. Il protocollo presentato è facilmente realizzabile e può essere utilizzato per la routine quotidiana in operazioni neurochirurgiche per evitare morbilità e migliorare il risultato postoperatorio. Il tempo supplementare per questo protocollo è inferiore a 9 min, che è significativamente più veloce rispetto ad altri protocolli 1,9,12,16. Riconoscendo il fatto che molti algoritmi sofisticati sono stati sviluppati recentemente le limita cartastesso per l&#39;utilizzo di un software disponibile in commercio e approvato dalla FDA. Tuttavia è obbligatorio per tener conto delle limitazioni di questa tecnica che sono menzionati sopra.

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			<td style="width:173px;">&#160;Signa LX version 9.1</td>
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			<td height="42" style="height:42px;width:216px;">Surgical Navigation System Srogram</td>
			<td style="width:71px;">Medtronic</td>
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			<td height="42" style="height:42px;width:216px;">Surgical Navigation System Srogram</td>
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			<td style="width:173px;">4500810331&#160; 20016318</td>
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</table>

dti of the visual pathway - white matter tracts and cerebral lesions

<html> DTI è una tecnica che identifica tratti di sostanza bianca (WMT) non invasiva in pazienti sani e non sani, utilizzando misure di diffusione. Simile a vie ottiche (VP), WMT non sono visibili con la risonanza magnetica classica o intra-operatoria con il microscopio. DIT aiuterà neurochirurghi per impedire la distruzione del VP, mentre la rimozione di lesioni adiacenti a questa WMT. Abbiamo effettuato DTI su cinquanta pazienti prima e dopo l&#39;intervento chirurgico tra il marzo 2012 al gennaio 2014 Per navigare abbiamo utilizzato una sequenza 3DT1 ponderato. Inoltre, abbiamo effettuato un T2-pesate e DTI-sequenze. I parametri utilizzati sono stati, FOV: 200 x 200 mm, spessore di strato: 2 mm, e matrice di acquisizione: 96 x 96 cedendo voxel quasi isotropico di 2 x 2 x 2 mm. Assiale MRI è stata effettuata utilizzando un gradiente di direzione 32 e uno b0-immagine. Abbiamo usato Echo-Planar-Imaging (EPI) e ASSET imaging parallelo con un fattore di accelerazione di 2 e B-valore di 800 s / mm ². Il tempo di scansione è inferiore a 9 min. ent &quot;&gt; I DTI-dati ottenuti sono stati elaborati utilizzando un programma approvato dalla FDA chirurgico sistema di navigazione che utilizza un approccio diretto fibra-tracking noto come l&#39;assegnazione di fibra da inseguimento continuo (FACT). Questa si basa sulla propagazione delle linee tra le regioni di interesse ( ROI) che è definito da un medico. un angolo massimo di 50, FA valore di 0,10 e ADC valore di arresto di 0,20 mm ² / s iniziare sono stati i parametri utilizzati per trattografia. Ci sono alcune limitazioni a questa tecnica. Il telaio limitato tempo di acquisizione impone compromessi nella qualità dell&#39;immagine. Un altro punto importante da non trascurare è il turno del cervello durante l&#39;intervento chirurgico. Per quanto riguarda la seconda risonanza magnetica intraoperatoria potrebbe essere utile. Inoltre, il rischio di falsi tratti positivi o falsi negativi deve essere preso in considerazione che potrebbero compromettere i risultati finali.

<html> Questo protocollo consente al medico di rappresentare adeguatamente le parti principali del VP. Può essere utilizzato con una piccola quantità di tempo al fine di evitare danni in pazienti con lesioni cerebrali prossimi alle aree eloquenti. Controlli postoperatori mostrano anche buoni risultati. VP è ritratto in figura 7 dopo che il paziente è stato operato da un glioblastoma. Figura 2 mostra il VP dopo recidiva di un glioblastoma. Gli autori riconoscono il fatto delle diffico...

Watch this Scientific Journal Video about DTI of the Visual Pathway - White Matter Tracts and Cerebral Lesions at JoVE.com

DTI of the Visual Pathway - White Matter Tracts and Cerebral Lesions

Diffusione tensore di imaging (DTI) è stata eseguita per provare a descrivere le parti principali del percorso visivo. L&#39;obiettivo era quello di utilizzare un farmaco approvato dalla workstation commerciale standard che potrebbe essere utilizzato per la routine di tutti i giorni per cercare di ridurre i danni post-operatoria del percorso visivo nei pazienti.

DTI della via visiva - White Matter Tracts e lesioni cerebrali

DTI is a technique that identifies white matter tracts (WMT) non-invasively in healthy and non-healthy patients using diffusion measurements. Similar to ...

dti-of-the-visual-pathway-white-matter-tracts-and-cerebral-lesions

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JoVE Journal

Medicine

13.9K Views.  Centre Hospitalier de Luxembourg.  Diffusione tensore di imaging (DTI) è stata eseguita per provare a descrivere le parti principali del percorso visivo. L'obiettivo era quello di utilizzare un farmaco approvato dalla workstation commerciale standard che potrebbe essere utilizzato per la routine di tutti i giorni per cercare di ridurre i danni post-operatoria del percorso visivo nei pazienti. 

Video: DTI of the Visual Pathway - White Matter Tracts and Cerebral Lesions

The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

Focal cerebral ischemia is among the most common type of stroke seen in patients. Due to the clinical significance there has been a prolonged effort to develop suitable animal models to study the events that unfold during ischemic insult. These techniques include transient or permanent, focal or global ischemia models using many different animal models, with the most common being rodents.
The permanent MCA ligation method which is also referred as pMCAo in the literature is used extensively as a focal ischemia model in rodents 1-6. This method was originally described for rats by Tamura et al. in 1981 7. In this protocol a craniotomy was used to access the MCA and the proximal regions were occluded by electrocoagulation. The infarcts involve mostly cortical and sometimes striatal regions depending on the location of the occlusion. This technique is now well established and used in many laboratories 8-13. Early use of this technique led to the definition and description of &#x201C;infarct core&#x201D; and &#x201C;penumbra&#x201D; 14-16, and it is often used to evaluate potential neuroprotective compounds 10, 12, 13, 17. Although the initial studies were performed in rats, permanent MCA ligation has been used successfully in mice with slight modifications 18-20 .
This model yields reproducible infarcts and increased post-survival rates. Approximately 80% of the ischemic strokes in humans happen in the MCA area 21 and thus this is a very relevant model for stroke studies. Currently, there is a paucity of effective treatments available to stroke patients, and thus there is a need for good models to test potential pharmacological compounds and evaluate physiological outcomes. This method can also be used for studying intracellular hypoxia response mechanisms in vivo. 
Here, we present the MCA ligation surgery in a C57/BL6 mouse. We describe the pre-surgical preparation, MCA ligation surgery and 2,3,5 Triphenyltetrazolium chloride (TTC) staining for quantification of infarct volumes.

Middle cerebral artery (MCA) ligation is a technique to study focal cerebral ischemia in animal models. In this method, the middle cerebral artery is exposed by craniotomy and ligated by cauterization. This method gives highly reproducible infarct volumes and increased post-operative survival rates compared to other methods available.

L&#39;applicazione di permanente legatura dell&#39;arteria cerebrale media nel mouse

Focal cerebral ischemia is among the most common type of stroke seen in patients. Due to the clinical significance there has been a prolonged effort to ...

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Medicina

Driving Simulation in the Clinic: Testing Visual Exploratory Behavior in Daily Life Activities in Patients with Visual Field Defects

Patients suffering from homonymous hemianopia after infarction of the posterior cerebral artery (PCA) report different degrees of constraint in daily life, despite similar visual deficits. We assume this could be due to variable development of compensatory strategies such as altered visual scanning behavior. Scanning compensatory therapy (SCT) is studied as part of the visual training after infarction next to vision restoration therapy. SCT consists of learning to make larger eye movements into the blind field enlarging the visual field of search, which has been proven to be the most useful strategy1, not only in natural search tasks but also in mastering daily life activities2. Nevertheless, in clinical routine it is difficult to identify individual levels and training effects of compensatory behavior, since it requires measurement of eye movements in a head unrestrained condition. Studies demonstrated that unrestrained head movements alter the visual exploratory behavior compared to a head-restrained laboratory condition3. Martin et al.4 and Hayhoe et al.5 showed that behavior demonstrated in a laboratory setting cannot be assigned easily to a natural condition. Hence, our goal was to develop a study set-up which uncovers different compensatory oculomotor strategies quickly in a realistic testing situation: Patients are tested in the clinical environment in a driving simulator. SILAB software (Wuerzburg Institute for Traffic Sciences GmbH (WIVW)) was used to program driving scenarios of varying complexity and recording the driver's performance. The software was combined with a head mounted infrared video pupil tracker, recording head- and eye-movements (EyeSeeCam, University of Munich Hospital, Clinical Neurosciences).
The positioning of the patient in the driving simulator and the positioning, adjustment and calibration of the camera is demonstrated. Typical performances of a patient with and without compensatory strategy and a healthy control are illustrated in this pilot study. Different oculomotor behaviors (frequency and amplitude of eye- and head-movements) are evaluated very quickly during the drive itself by dynamic overlay pictures indicating where the subjects gaze is located on the screen, and by analyzing the data. Compensatory gaze behavior in a patient leads to a driving performance comparable to a healthy control, while the performance of a patient without compensatory behavior is significantly worse. The data of eye- and head-movement-behavior as well as driving performance are discussed with respect to different oculomotor strategies and in a broader context with respect to possible training effects throughout the testing session and implications on rehabilitation potential.

Patients with visual deficits after stroke report about different constraints in daily life most likely due to variable compensatory strategies, which are difficult to differentiate in clinical routine. We present a clinical set-up which allows measurement of different compensatory head- and eye-movement-strategies and evaluating their effects on driving performance.

Simulazione di guida presso la Clinica: Test di Visual comportamento esplorativo in attività della vita quotidiana in pazienti con difetti del campo visivo

Patients suffering from homonymous hemianopia after infarction of the posterior cerebral artery (PCA) report different degrees of constraint in daily ...

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

Sistema ottimizzato per il monitoraggio perfusione cerebrale nel modello Rat Stroke of intraluminale occlusione dell&#39;arteria cerebrale media

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow (ischemia) 1. Middle cerebral artery (MCA) delivers a great majority of blood to the lateral surface of the cortex 2, is the most common site of human stroke 3, and ischemia within its territory can result in extensive dysfunction or death 1,4,5. Survivors of ischemic stroke often suffer loss or disruption of motor capabilities, sensory deficits, and infarct. In an effort to capture these key characteristics of stroke, and thereby develop effective treatment, a great deal of emphasis is placed upon animal models of ischemia in MCA.
Here we present a method of permanently occluding a cortical surface blood vessel. We will present this method using an example of a relevant vessel occlusion that models the most common type, location, and outcome of human stroke, permanent middle cerebral artery occlusion (pMCAO). In this model, we surgically expose MCA in the adult rat and subsequently occlude via double ligature and transection of the vessel. This pMCAO blocks the proximal cortical branch of MCA, causing ischemia in all of MCA cortical territory, a large portion of the cortex. This method of occlusion can also be used to occlude more distal portions of cortical vessels in order to achieve more focal ischemia targeting a smaller region of cortex. The primary disadvantages of pMCAO are that the surgical procedure is somewhat invasive as a small craniotomy is required to access MCA, though this results in minimal tissue damage. The primary advantages of this model, however, are: the site of occlusion is well defined, the degree of blood flow reduction is consistent, functional and neurological impairment occurs rapidly, infarct size is consistent, and the high rate of survival allows for long-term chronic assessment.

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

We describe a highly reproducible method for the permanent occlusion of a rodent major cerebral blood vessel. This technique can be accomplished with very little peripheral damage, minimal blood loss, a high rate of long-term survival, and consistent infarct volume commensurate with the human clinical population.

Cerebrale permanente occlusione di vasi<em&gt; Via</em&gt; Doppio Legatura e resezione

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow ...

Generation and 3-Dimensional Quantitation of Arterial Lesions in Mice Using Optical Projection Tomography

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating important information on the pathogenesis of lesion formation and the action of novel therapies. Use of atherosclerosis-prone mice, surgical methods of lesion induction, and dietary modification has dramatically improved understanding of the mechanisms that contribute to disease development and the potential of new treatments.
Classically, analysis of lesions is performed ex vivo using 2-dimensional histological techniques. This article describes application of optical projection tomography (OPT) to 3-dimensional quantitation of arterial lesions. As this technique is non-destructive, it can be used as an adjunct to standard histological and immunohistochemical analyses.
Neointimal lesions were induced by wire-insertion or ligation of the mouse femoral artery whilst atherosclerotic lesions were generated by administration of an atherogenic diet to apoE-deficient mice.
Lesions were examined using OPT imaging of autofluorescent emission followed by complementary histological and immunohistochemical analysis. OPT clearly distinguished lesions from the underlying vascular wall. Lesion size was calculated in 2-dimensional sections using planimetry, enabling calculation of lesion volume and maximal cross-sectional area. Data generated using OPT were consistent with measurements obtained using histology, confirming the accuracy of the technique and its potential as a complement (rather than alternative) to traditional methods of analysis.
This work demonstrates the potential of OPT for imaging atherosclerotic and neointimal lesions. It provides a rapid, much needed ex vivo technique for the routine 3-dimensional quantification of vascular remodelling.

Ex vivo analysis of arterial lesions from animal models of cardiovascular disease classically relies on histological and immunohistochemical techniques. These provide 2-dimensional measurements in 3-dimensional lesions. This manuscript describes the generation of arterial lesions for quantitative analysis in 3-dimensions using optical projection tomography.

Generazione e quantificazione 3-Dimensional di arteriosa lesioni nei topi mediante tomografia ottica di proiezione

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating ...

Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have been developed to facilitate the study of the molecular and cellular pathophysiology of this disease. In this manuscript we describe specific techniques for the quantification and characterization of atherosclerotic lesions in the murine aortic sinus and ascending aorta. The advantage of this procedure is that it provides an accurate measurement of the cross-sectional area and total volume of the lesion, which can be used to compare atherosclerotic progression across different treatment groups. This is possible through the use of the valve leaflets as an anatomical landmark, together with careful adjustment of the sectioning angle. We also describe basic staining methods that can be used to begin to characterize atherosclerotic progression. These can be further modified to investigate antigens of specific interest to the researcher. The described techniques are generally applicable to a wide variety of existing and newly created dietary and genetically-induced models of atherogenesis.

We describe procedures to quantify and characterize atherosclerotic lesions in mouse models using precision sectioning of the aortic sinus and ascending aorta combined with histochemical and immunohistochemical analysis.

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have ...

Dynamic Visual Tests to Identify and Quantify Visual Damage and Repair Following Demyelination in Optic Neuritis Patients

In order to follow optic neuritis patients and evaluate the effectiveness of their treatment, a handy, accurate and quantifiable tool is required to assess changes in myelination at the central nervous system (CNS). However, standard measurements, including routine visual tests and MRI scans, are not sensitive enough for this purpose. We present two visual tests addressing dynamic monocular and binocular functions which may closely associate with the extent of myelination along visual pathways. These include Object From Motion (OFM) extraction and Time-constrained stereo protocols. In the OFM test, an array of dots compose an object, by moving the dots within the image rightward while moving the dots outside the image leftward or vice versa. The dot pattern generates a camouflaged object that cannot be detected when the dots are stationary or moving as a whole. Importantly, object recognition is critically dependent on motion perception. In the Time-constrained Stereo protocol, spatially disparate images are presented for a limited length of time, challenging binocular 3-dimensional integration in time. Both tests are appropriate for clinical usage and provide a simple, yet powerful, way to identify and quantify processes of demyelination and remyelination along visual pathways. These protocols may be efficient to diagnose and follow optic neuritis and multiple sclerosis patients.
In the diagnostic process, these protocols may reveal visual deficits that cannot be identified via current standard visual measurements. Moreover, these protocols sensitively identify the basis of the currently unexplained continued visual complaints of patients following recovery of visual acuity. In the longitudinal follow up course, the protocols can be used as a sensitive marker of demyelinating and remyelinating processes along time. These protocols may therefore be used to evaluate the efficacy of current and evolving therapeutic strategies, targeting myelination of the CNS.

Object From Motion (OFM) and Time-constrained stereo protocols are sensitive tools to identify monocular and binocular dynamic visual function deficits, which are uniquely affected in optic neuritis patients. Furthermore, these tests may be used as quantitative noninvasive tools to assess the extent of myelination along visual pathways.

I test visivi dinamici per identificare e quantificare eventuali danni visivi e riparazione seguito demielinizzazione nella neurite ottica pazienti

In order to follow optic neuritis patients and evaluate the effectiveness of their treatment, a handy, accurate and quantifiable tool is required to ...

A Versatile Murine Model of Subcortical White Matter Stroke for the Study of Axonal Degeneration and White Matter Neurobiology

Stroke affecting white matter accounts for up to 25% of clinical stroke presentations, occurs silently at rates that may be 5-10 fold greater, and contributes significantly to the development of vascular dementia. Few models of focal white matter stroke exist and this lack of appropriate models has hampered understanding of the neurobiologic mechanisms involved in injury response and repair after this type of stroke. The main limitation of other subcortical stroke models is that they do not focally restrict the infarct to the white matter or have primarily been validated in non-murine species. This limits the ability to apply the wide variety of murine research tools to study the neurobiology of white matter stroke. Here we present a methodology for the reliable production of a focal stroke in murine white matter using a local injection of an irreversible eNOS inhibitor. We also present several variations on the general protocol including two unique stereotactic variations, retrograde neuronal tracing, as well as fresh tissue labeling and dissection that greatly expand the potential applications of this technique. These variations allow for multiple approaches to analyze the neurobiologic effects of this common and understudied form of stroke.

Here we present methodology for the production of a focal stroke in murine white matter by local injection of an irreversible endothelial nitric oxide synthase (eNOS) inhibitor (L-Nio). Presented are two stereotactic variations, retrograde neuronal tracing, and fresh tissue labeling and dissection that expand the potential applications of this technique.

A Corsa Versatile Modello murino di Subcortical bianco Materia per lo Studio della assonale degenerazione e bianco Materia Neurobiologia

Stroke affecting white matter accounts for up to 25% of clinical stroke presentations, occurs silently at rates that may be 5-10 fold greater, and ...

Autologous Microfractured and Purified Adipose Tissue for Arthroscopic Management of Osteochondral Lesions of the Talus

In recent years, regenerative techniques have been increasingly studied and used to treat osteochondral lesions of the talus. In particular, several studies have focused their attention on mesenchymal stem cells derived from adipose tissue. Adipose-derived stem cells (ADSCs) exhibit morphological characteristics and properties similar to other mesenchymal cells, and are able to differentiate into several cellular lines. Moreover, these cells are also widely available in the subcutaneous tissue, representing 10 - 30% of the normal body weight, with a concentration of 5,000 cells per gram of tissue.
In the presented technique, the first step involves harvesting ADSCs from the abdomen and a process of microfracture and purification; next, the surgical procedure is performed entirely arthroscopically, with less soft tissue dissection, better joint visualization, and a faster recovery compared with standard open procedures. Arthroscopy is characterized by a first phase in which the lesion is identified, isolated, and prepared with microperforations; the second step, performed dry, involves injection of adipose tissue at the level of the lesion.
Between January 2016 and September 2016, four patients underwent arthroscopic treatment of osteochondral lesion of the talus with microfractured and purified adipose tissue. All patients reported clinical improvement six months after surgery with no reported complications. Functional scores at the latest follow-up are encouraging and confirm that the technique provides reliable pain relief and improvements in patients with osteochondral lesion of the talus.

The aim of this study is to report a protocol for the arthroscopic treatment of osteochondral lesions of the talus using microfractured and purified adipose-derived stem cells.

Microfratturate autologo e nel tessuto adiposo purificato per l'amministrazione artroscopica di lesioni osteocondrali dell'astragalo

In recent years, regenerative techniques have been increasingly studied and used to treat osteochondral lesions of the talus. In particular, several ...

Assessment of Static Graviceptive Perception in the Roll-Plane using the Subjective Visual Vertical Paradigm

Vestibular disorders are among the most common syndromes in medicine. In recent years, new vestibular diagnostic systems have been introduced that allow the examination of all semicircular canals in the clinical setting. Assessment methods of the otolithic system, which is responsible for the perception of linear acceleration and perception of gravity, are far less in clinical use. There are several experimental approaches for measuring the perception of gravity. The most frequently used method is the determination of the subjective visual vertical. This is usually measured with the head in an upright position. We present here an assessment method for testing otolith function in the roll plane. The subjective visual vertical is measured in the head upright position as well as with head inclination of &#177; 15&#176; and &#177; 30&#176; in the roll plane. This extended functional paradigm is an easy-to-perform clinical test of otolith function and ensures increased information content for the detection of impaired graviceptive perception.

The perception of gravity is commonly determined by the subjective visual vertical in the head upright position. The additional assessment at head tilts of &#177; 15&#176; and &#177; 30&#176; in the roll plane ensures increased information content for the detection of impaired graviceptive perception.

Valutazione della percezione gravicettiva statica nel Roll-Plane utilizzando il paradigma verticale visivo soggettivo

Vestibular disorders are among the most common syndromes in medicine. In recent years, new vestibular diagnostic systems have been introduced that allow ...