Alcune delle figure in questo manoscritto sono stati generosamente fornito dal laboratorio di Bart O. Williams. Gli autori sono supportati dal National Cancer Institute concede U54CA143803, CA163124, CA093900, CA143055 E.

Questo protocollo incontra e segue le linee guida stabilite dal Comitato Istituzionale cura degli animali e Usa Johns Hopkins University. 1. Micro-dissezione una prostata mouse <ol><li> Eutanasia il mouse per asfissia anidride carbonica o un metodo approvato alternativo in conformità con le linee guida per la cura degli animali e uso istituzionale. </li><li> Pin il mouse nella posizione supina ad una tavola di dissezione mettendo un perno attraverso ciascuno dei quattro zampe. </li><li> Spruzzare la addome con etanolo al 70%. </li><li> Tenere la pelle del basso addome con una pinza, e usare le forbici per tagliare attraverso la pelle e peritoneo circa 1 cm anteriormente all&#39;apertura del pene, attenzione a non tagliare tutti gli organi. </li><li> Tagliare la pelle e del peritoneo dal basso addome, fino entrambi i lati dell&#39;addome alla gabbia toracica, esponendo la cavità peritoneale. </li><li> Identificare la vescica e del tratto urogenitale, ed esporli delicatamente spostando grassi e altri organi di lato. </li><li> Usiforcipe Ng, presa vasi deferenti alla base vicino l&#39;uretra, e strappare via. </li><li> Ripetere con il dotto deferente opposte. </li><li> Utilizzando pinze, accuratamente afferrare la vescica e tirarlo, mentre simultaneamente con le forbici per tagliare attraverso l&#39;uretra sotto la vescica e la prostata ventrale (circa 1 cm sotto la base della vescica). Nota: Poiché la vescica è tirato su, il (vescica contenente una sezione dell&#39;uretra, tutti i lobi della prostata e le vescicole seminali) intero tratto urogenitale verrà con esso. Ci può essere una certa resistenza. </li><li> Posizionare il tratto urogenitale (UGT) in un 60-100 mm piastra di Petri contenente circa 5-10 ml tampone fosfato salino (PBS). </li><li> Eseguire la parte restante del protocollo sotto un microscopio di dissezione. Utilizzare due pinza sottile, uno in ogni mano. Tenere le pinze &quot;come una matita&quot; e riposare gli avambracci sul banco in modo da stabilizzare loro. </li><li> Utilizzare una pinza sottile per posizionare la UGT in modo tale che la vescica èsopra, l&#39;uretra è puntato verso il basso, e le vescicole seminali sono posizionate su entrambi i lati della vescica. </li><li> Grip l&#39;uretra con una pinza ed estrarre delicatamente il grasso con gli altri pinze senza strappare via qualsiasi tessuto prostatico. Nota: Fat apparirà relativo &quot;lucido&quot; al tessuto circostante. </li><li> Una volta che il grasso viene rimosso, utilizzare pinze per strappare il tessuto connettivo tra i due lobi ventrali della prostata e separarli. </li><li> Per rimuovere uno dei lobi ventrali, usare una pinza per afferrare un unico lobo ventrale, vicino alla punta, e utilizzare le altre pinze per afferrare che lobo alla base il più vicino al uretra più possibile. Mentre ancora presa alla base del lobo, utilizzare le altre pinze per afferrare la uretra. Estrarre il lobo di distanza dall&#39;uretra con una ditta, movimento regolare. Assicurarsi che nessun tessuto prostatico rimane attaccato al uretra. Posizionare il lobo nel mezzo appropriata, a seconda del passo successivo sperimentale. <ol><li> Per risolvere il problema e di paraffina-incorporare le sampio per l&#39;analisi istologica, posizionare il lobo in formalina neutra tamponata al 10% (NBF) 8. </li><li> Posizionare il lobo di congelamento di media, come ad esempio Office, per la crioconservazione e sezionamento 8. </li><li> Posizionare il lobo in PBS freddo per l&#39;isolamento delle cellule unico per la cultura o citometria a flusso 9. </li></ol></li><li> Ripetere il passaggio 1.15 con l&#39;altro lobo ventrale. </li><li> Per rimuovere uno dei lobi frontali, usare il forcipe per tirare delicatamente il lobo di distanza dal vescicole seminali, facendo attenzione a non forare la vescicole seminali. Una volta che il lobo è indipendente della vescicola seminale, rimuoverlo nello stesso modo come il lobo ventrale passo 1.15. </li><li> Ripetere il passaggio 1.17 con l&#39;altro lobo anteriore. </li><li> Vibrazione tratto urogenitale restante sopra tale che la prostata dorsale è visibile. </li><li> Per rimuovere uno dei lobi dorsolaterali, utilizzare pinze per strappare il tessuto connettivo tra i due lobi dorsolaterali e anche il tessuto connettivo tra i lobi e tegli posteriormente regione delle vescicole seminali. Rimuovere il lobo in modo simile come il lobo ventrale passo 1.15. </li><li> Ripetere il passaggio 1.20 con l&#39;altro lobo dorsolaterale. </li></ol> 2. castrazione chirurgica Nota: Si tratta di un intervento chirurgico di sopravvivenza, così asepsi, anestesia, e la gestione del dolore sono importanti per il completamento etica e di successo di questo protocollo. Seguire tutte le linee guida per la cura degli animali e uso istituzionale. Utilizzare una fonte di calore esterna, ad esempio una coperta di acqua ricircolo, durante la procedura chirurgica per prevenire l&#39;ipotermia. Non lasciare un animale incustodito fino a quando non ha ripreso conoscenza sufficiente a mantenere decubito sternale. Non restituire un animale che ha subito un intervento chirurgico per la compagnia di altri animali fino alla completa guarigione. Autoclavare tutti gli strumenti chirurgici prima dell&#39;uso in chirurgia. <ol><li> Su una superficie pulita e sterile anestetizzare il mouse utilizzando misure appropriate (ad esempio 2% isoflurano per via inalatoria) e posizionare il mOuse in posizione supina. Nota: Metodi anestesia alternativi, come la chetamina, possono anche essere utilizzati, a seconda del protocollo animali approvato. </li><li> Controllare i riflessi del topo pizzicando le dita dei piedi. Se il mouse non reagisce, passare alla fase successiva. Se non reagisce, aspettare 1-3 minuti e prova di nuovo. Radere l&#39;area chirurgica con un rasoio elettrico. </li><li> Asetticamente preparare l&#39;addome del mouse utilizzando alternati scrub di etanolo al 70% e iodio o procedure raccomandate dal personale veterinario istituzionale o IACUC. </li><li> Usando un bisturi sterile, praticare un&#39;incisione verticale 1 centimetro attraverso la pelle sulla linea mediana del basso addome, circa 1,5 cm anteriormente al pene. </li><li> Fai una piccola incisione (&lt;1 cm) attraverso il peritoneo, attenzione a non tagliare tutti gli organi. </li><li> Utilizzando pinze sterili, afferrare il bordo tagliato del peritoneo e sollevarlo leggermente in modo da poter vedere la cavità peritoneale sotto. </li><li> Utilizzando un altro pinza sterile, raggiungere in the cavità peritoneale e il grip sia il cuscinetto di grasso testicolare a sinistra oa destra, laterale alla vescica. Estrarre il cuscinetto adiposo attraverso l&#39;apertura nel peritoneo e pelle fino testicolo esce pure. Fare attenzione a non danneggiare altri tessuti o organi durante questa fase. </li><li> Usando una penna cauterizzazione, tagliare il cuscinetto adiposo che sta tenendo il testicolo. Tagliare lentamente attraverso l&#39;arteria testicolare con la penna cauterizzazione Nota: Se l&#39;arteria testicolare è tagliato troppo veloce, ci può essere troppo sanguinamento e il mouse possono avere per essere eutanasia. </li><li> Una volta che il cuscinetto di grasso e l&#39;arteria sono tagliati, rimuovere il pad testicoli e grassi. </li><li> Ripetere i passaggi 2,6-2,9 con l&#39;altro testicolo. Nota: Un metodo alternativo per la rimozione dei testicoli è quello di legare chirurgicamente l&#39;arteria testicolare, quindi tagliato nave con le forbici. </li><li> Sutura del peritoneo chiusa con suture assorbibili. </li><li> Staple la pelle chiusa con clip ferita chirurgica. </li><li> Somministrare un analgesico per gestire il dolore, e il luogoil mouse in una gabbia pulita su una gabbia caldo per 5-10 min. Monitorare il mouse per i segni di dolore o sanguinamento. </li></ol>

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S., Labrie, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unique+features+of+the+basal+cells+of+human+prostate+epithelium.">Unique features of the basal cells of human prostate epithelium.</a> Microsc Res Tech. 51 (5), 436-446 (2000).</li><li>Prins, G. S., Huang, L., Birch, L., Pu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+estrogens+in+normal+and+abnormal+development+of+the+prostate+gland.">The role of estrogens in normal and abnormal development of the prostate gland.</a> Ann N Y Acad Sci. 1089, 1-13 (2006).</li><li>Valkenburg, K. 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Gli autori dichiarano di non avere interessi finanziari concorrenti.

Prostate micro-dissezione consente di sperimentazione specifici lobi e analisi della prostata mouse (Figura 1). Nei modelli di topo geneticamente, fenotipi possono essere visti in un lobo che non si vede in altri. Inoltre, per l&#39;analisi istologica, questo protocollo assicura che la massima quantità di tessuto prostatico puro può essere sezionato e macchiato, senza altri tessuti del tratto urogenitale presenti nella sezione. Infine, per gli esperimenti a cella singola, questo protocollo permette pe...

La prostata è il sito più comune di cancro negli uomini negli Stati Uniti. Quasi 220.000 uomini ogni anno saranno con diagnosi di cancro alla prostata, e circa 27.000 uomini soccombere alla loro malattia 1. Gli uomini hanno un rischio 1 a 7 vita di una diagnosi di cancro alla prostata negli Stati Uniti 1. Iperplasia prostatica benigna (BPH), un noncancerous ingrandimento legata all&#39;età della prostata, è anche una condizione molto diffusa, che colpisce oltre l&#39;80% degli uomini oltre 80 2. In quanto tale, la prostata è al centro di molte ricerche. Modelli di topo sono stati utilizzati ampiamente per studiare le malattie della prostata 3. Nel complesso, la prostata mouse è un ottimo rappresentante della ghiandola umana 4, ma ci sono somiglianze e le differenze tra il mouse e anatomia della prostata umana e fisiologia 5. In entrambe le specie, la prostata si trova alla base della vescica, che circonda l&#39;uretra. La prostata di essere una sola loessere, diviso in quattro zone: centrale, di transizione, periferiche, e stroma fibromuscolare anteriore. Al contrario, la prostata mouse è suddiviso in tre lobi accoppiati a diverse posizioni intorno all&#39;uretra: anteriore, ventrale e dorsolateral. A livello cellulare, la differenza principale è che la cellula basale rapporto cellulare luminale nell&#39;uomo è di circa 1: 1, ma è 1: 4 in topi 6. Lo stroma murino è inoltre differente nei topi relativi agli esseri umani - esseri umani hanno più estesa muscolatura liscia, mentre lo strato muscolare è molto più sottile nella prostata murino. La corretta identificazione, la dissezione, e la manipolazione della prostata del mouse sono essenziali per la ricerca del mouse prostata. i livelli ormonali influenzano drasticamente lo sviluppo e l&#39;omeostasi della ghiandola prostatica in entrambi gli uomini e topi. Mentre gli estrogeni sembra giocare un ruolo nello sviluppo della prostata 7, l&#39;ormone più importante nella prostata è testosterone. Il testosterone è essenziale per uno sviluppo ghiandolared manutenzione. Dopo castrazione fisica o chimica, circa il 90% delle cellule luminali del Apoptose adulti prostata, e la ghiandola restringe. Se il testosterone viene reintrodotto ad un individuo in condizioni di castrazione, la prostata è in grado di rigenerarsi a piena capacità. Il testosterone sembra anche a guidare il cancro alla prostata, che è il motivo per cui la terapia di deprivazione androgenica è una strategia terapeutica di uso comune. Tuttavia, molti tumori della prostata diventano resistenti alla deprivazione androgenica. Inoltre, gli uomini sottoposti a terapia di deprivazione androgenica per il trattamento del cancro alla prostata sottoposti a cicli di condizioni castrati e rigenerati. Nel topo, la castrazione chirurgica, insieme con la rigenerazione ormonale della prostata reintroducendo il testosterone, è un importante strumento con cui studiare la resistenza castrazione e gli effetti del testosterone in bicicletta sulla prostata. In questo articolo, si discuterà e dimostrare le tecniche adeguate in base ai quali Locatee micro-sezionare una prostata mouse, nonché chirurgicamente castrare un mouse.

<table border="0" cellpadding="0" cellspacing="0" width="741">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Surgical tools</td>
			<td style="width:145px;">Roboz</td>
			<td style="width:113px;">Various</td>
			<td style="width:239px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Formaldehyde</td>
			<td>Sigma</td>
			<td>F8775</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">OCT medium</td>
			<td>VWR</td>
			<td>25608-930</td>
			<td style="width:239px;">Must be placed on dry ice to freeze, and stored at -80C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PBS</td>
			<td>Life Technologies</td>
			<td align="right">10010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane</td>
			<td>Johns Hopkins</td>
			<td></td>
			<td>Also available from vendors online</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cautery Pen</td>
			<td>Medline</td>
			<td>ESCT002</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Silk suture</td>
			<td>AD Surgical</td>
			<td>M-S330R19</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical Wound Clips</td>
			<td>Roboz</td>
			<td>RS-9265</td>
			<td></td>
		</tr>
	</tbody>
</table>

murine prostate micro-dissection and surgical castration

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has been used as a surrogate for discoveries in human prostate development and disease. Prostate micro-dissection allows consistent study of lobe-specific prostate anatomy, histology, and cellular characteristics in the absence of contamination of other tissues. Testosterone affects prostate development and disease. Androgen deprivation therapy is a common treatment for prostate cancer patients, but many prostate tumors become castration-resistant. Surgical castration of mouse models allows for the study of castration resistance and other facets of hormonal biology on the prostate. This procedure can be coupled with testosterone reintroduction, or hormonal regeneration of the prostate, a powerful method to study stem cell lineages in the prostate. Together, prostate micro-dissection and surgical castration opens up a multitude of opportunities for robust and consistent research of prostate development and disease. This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse.

Tutti i sei lobi della prostata sono stati rimossi da un mouse via prostata micro-dissezione (Figura 1). Il tratto urogenitale completa (UGT) è composta da tutti i lobi della prostata, della vescica, vescicole seminali, e l&#39;uretra (Figura 1A). I vasi deferenti attribuisce al uretra, ma non è necessaria per la prostata micro-dissezione, e possono quindi essere staccati prima della rimozione della UGT tagliando l&#39;uretra (Figura 1b-1c).</s...

Watch this Scientific Journal Video about Murine Prostate Micro-dissection and Surgical Castration at JoVE.com

Murine Prostate Micro-dissection and Surgical Castration

Questo manoscritto descrive i protocolli per la prostata micro-dissezione e la castrazione chirurgica nel topo di laboratorio. Noi raffiguriamo anche risultati rappresentativi prodotti da questi protocolli. Infine, si discute i vantaggi e l&#39;utilizzo di questi protocolli.

Murino della prostata Micro-dissezione e la castrazione chirurgica

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has ...

murine-prostate-micro-dissection-and-surgical-castration

Research

JoVE Journal

Medicine

45.2K Views.  Johns Hopkins University.  Questo manoscritto descrive i protocolli per la prostata micro-dissezione e la castrazione chirurgica nel topo di laboratorio. Noi raffiguriamo anche risultati rappresentativi prodotti da questi protocolli. Infine, si discute i vantaggi e l'utilizzo di questi protocolli. 

Video: Murine Prostate Micro-dissection and Surgical Castration

Microvascular Decompression: Salient Surgical Principles and Technical Nuances

Trigeminal neuralgia is a disorder associated with severe episodes of lancinating pain in the distribution of the trigeminal nerve. Previous reports indicate that 80-90% of cases are related to compression of the trigeminal nerve by an adjacent vessel. The majority of patients with trigeminal neuralgia eventually require surgical management in order to achieve remission of symptoms. Surgical options for management include ablative procedures (e.g., radiosurgery, percutaneous radiofrequency lesioning, balloon compression, glycerol rhizolysis, etc.) and microvascular decompression. Ablative procedures fail to address the root cause of the disorder and are less effective at preventing recurrence of symptoms over the long term than microvascular decompression. However, microvascular decompression is inherently more invasive than ablative procedures and is associated with increased surgical risks. Previous studies have demonstrated a correlation between surgeon experience and patient outcome in microvascular decompression. In this series of 59 patients operated on by two neurosurgeons (JSN and PEK) since 2006, 93% of patients demonstrated substantial improvement in their trigeminal neuralgia following the procedure&#x2014;with follow-up ranging from 6 weeks to 2 years. Moreover, 41 of 66 patients (approximately 64%) have been entirely pain-free following the operation.
In this publication, video format is utilized to review the microsurgical pathology of this disorder. Steps of the operative procedure are reviewed and salient principles and technical nuances useful in minimizing complications and maximizing efficacy are discussed.

There are many available options for management of the patient with trigeminal neuralgia. Microvascular decompression, while the most invasive of all options, is also the most effective at achieving long term remission of symptoms. Video instruction on how to maximize efficacy and minimize complications with this procedure is described.

La decompressione microvascolare: Principi salienti chirurgici e sfumature tecniche

Trigeminal neuralgia is a disorder associated with severe episodes of lancinating pain in the distribution of the trigeminal nerve. Previous reports ...

Ricerca

Medicina

Evaluation of Nanoparticle Uptake in Tumors in Real Time Using Intravital Imaging

Current technologies for tumor imaging, such as ultrasound, MRI, PET and CT, are unable to yield high-resolution images for the assessment of nanoparticle uptake in tumors at the microscopic level1,2,3, highlighting the utility of a suitable xenograft model in which to perform detailed uptake analyses. Here, we use high-resolution intravital imaging to evaluate nanoparticle uptake in human tumor xenografts in a modified, shell-less chicken embryo model. The chicken embryo model is particularly well-suited for these in vivo analyses because it supports the growth of human tumors, is relatively inexpensive and does not require anesthetization or surgery 4,5. Tumor cells form fully vascularized xenografts within 7 days when implanted into the chorioallantoic membrane (CAM) 6. The resulting tumors are visualized by non-invasive real-time, high-resolution imaging that can be maintained for up to 72 hours with little impact on either the host or tumor systems. Nanoparticles with a wide range of sizes and formulations administered distal to the tumor can be visualized and quantified as they flow through the bloodstream, extravasate from leaky tumor vasculature, and accumulate at the tumor site. We describe here the analysis of nanoparticles derived from Cowpea mosaic virus (CPMV) decorated with near-infrared fluorescent dyes and/or polyethylene glycol polymers (PEG) 7, 8, 9,10,11. Upon intravenous administration, these viral nanoparticles are rapidly internalized by endothelial cells, resulting in global labeling of the vasculature both outside and within the tumor7,12. PEGylation of the viral nanoparticles increases their plasma half-life, extends their time in the circulation, and ultimately enhances their accumulation in tumors via the enhanced permeability and retention (EPR) effect 7, 10,11. The rate and extent of accumulation of nanoparticles in a tumor is measured over time using image analysis software. This technique provides a method to both visualize and quantify nanoparticle dynamics in human tumors.

We present a novel approach to quantify nanoparticle localization in the vasculature of human xenografted tumors using dynamic, real-time intravital imaging in an avian embryo model.

Valutazione della captazione nanoparticelle nei tumori in tempo reale utilizzando intravitale Imaging

Current technologies for tumor imaging, such as ultrasound, MRI, PET and CT, are unable to yield high-resolution images for the assessment of nanoparticle ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

Our laboratory has developed a novel orthotopic implantation model of human prostate cancer (PCa). As PCa death is not due to the primary tumor, but rather the formation of distinct metastasis, the ability to effectively model this progression pre-clinically is of high value. In this model, cells are directly implanted into the ventral lobe of the prostate in Balb/c athymic mice, and allowed to progress for 4-6 weeks. At experiment termination, several distinct endpoints can be measured, such as size and molecular characterization of the primary tumor, the presence and quantification of circulating tumor cells in the blood and bone marrow, and formation of metastasis to the lung. In addition to a variety of endpoints, this model provides a picture of a cells ability to invade and escape the primary organ, enter and survive in the circulatory system, and implant and grow in a secondary site. This model has been used effectively to measure metastatic response to both changes in protein expression as well as to response to small molecule therapeutics, in a short turnaround time.

This orthotopic model of human prostate cancer allows for quantification of tumor size, circulating tumor cells, and formation of distinct metastasis to the lung. As cells must escape the primary organ, enter the blood stream, and implant into a secondary site, this model effectively recapitulates the scenario in humans.

Un ortotopico Modello murino di Human Prostate Cancer Metastasi

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Surgical Fixation of Sternal Fractures: Preoperative Planning and a Safe Surgical Technique Using Locked Titanium Plates and Depth Limited Drilling

Different ways to stabilize a sternal fracture are described in literature. Respecting different mechanisms of trauma such as the direct impact to the anterior chest wall or the flexion-compression injury of the trunk, there is a need to retain each sternal fragment in the correct position while neutralizing shearing forces to the sternum. Anterior sternal plating provides the best stability and is therefore increasingly used in most cases. However, many surgeons are reluctant to perform sternal osteosynthesis due to possible complications such as difficulties in preoperative planning, severe injuries to mediastinal organs, or failure of the performed method.
This manuscript describes one possible safe way to stabilize different types of sternal fractures in a step by step guidance for anterior sternal plating using low profile locking titanium plates. Before surgical treatment, a detailed survey of the patient and a three dimensional reconstructed computed tomography is taken out to get detailed information of the fracture&#8217;s morphology. The surgical approach is usually a midline incision. Its position can be described by measuring the distance from upper sternal edge to the fracture and its length can be approximated by the summation of 60 mm for the basis incision, the thickness of presternal soft tissue and the greatest distance between the fragments in case of multiple fractures.
Performing subperiosteal dissection along the sternum while reducing the fracture, using depth limited drilling, and fixing the plates prevents injuries to mediastinal organs and vessels.
Transverse fractures and oblique fractures at the corpus sterni are plated longitudinally, whereas oblique fractures of manubrium, sternocostal separation and any longitudinally fracture needs to be stabilized by a transverse plate from rib to sternum to rib. Usually the high convenience of a patient is seen during follow up as well as a precise reconstruction of the sternal morphology.

Here we present a method to stabilize sternal fractures by using locked titanium plates in a low profile design. Performing subperiosteal dissection along the sternum while reducing the fracture, using depth limited drilling, and fixing the plates provides a safe surgical way.

Fissazione chirurgica di fratture sterno: la pianificazione preoperatoria e una tecnica chirurgica sicuro con piastre di titanio bloccati e limitata profondità di foratura

Different ways to stabilize a sternal fracture are described in literature. Respecting different mechanisms of trauma such as the direct impact to the ...

A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the exact pathophysiology remains unknown. In order to unravel the pathophysiology a murine model of AVF-failure was developed in which the configuration of the anastomosis resembles the preferred situation in the clinical setting. A model was described in which an AVF is created by connecting the venous end of the branch of the external jugular vein to the side of the common carotid artery using interrupted sutures. At a histological level, we observed progressive stenotic intimal lesions in the venous outflow tract that is also seen in failed human AVFs. Although this procedure can be technically challenging due to the small dimensions of the animal, we were able to achieve a surgical success rate of 97% after sufficient training. The key advantage of a murine model is the availability of transgenic animals. In view of the different proposed mechanisms that are responsible for AVF failure, disabling genes that might play a role in vascular remodeling can help us to unravel the complex pathophysiology of AVF failure.

Here we present a murine model of arteriovenous fistula (AVF) failure in which a clinically relevant anastomotic configuration is incorporated. This model can be used to study the pathophysiology and to test possible therapeutic interventions.

A Novel Modello murino di fistola artero-venosa Fallimento: la procedura chirurgica in dettaglio

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information about this disease. The human orthotopic prostate cancer xenograft mouse model provides a useful alternative approach for understanding the specific interactions between genetically and molecularly altered tumor cells, their organ microenvironment, and for evaluation of efficacy of therapeutic regimens. This is a well characterized model designed to study the molecular events of primary tumor development and it recapitulates the early events in the metastatic cascade prior to embolism and entry of tumor cells into the circulation. Thus it allows elucidation of molecular mechanisms underlying the initial phase of metastatic disease. In addition, this model can annotate drug targets of clinical relevance and is a valuable tool to study prostate cancer progression. In this manuscript we describe a detailed procedure to establish a human orthotopic prostate cancer xenograft mouse model.

Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

Pre-clinica ortotopico Modello murino di Human cancro alla prostata

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Intravital Microscopy of Tumor-associated Vasculature Using Advanced Dorsal Skinfold Window Chambers on Transgenic Fluorescent Mice

Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static information and is often not sufficient for a correct interpretation. Intravital evaluation, in which a process is followed in time, provides extra and often unexpected information. With the creation of transgenic animals expressing cell-specific markers and live cell tracers, improvements to imaging equipment, and the development of several imaging chambers, intravital microscopy has become an important tool to better understand biological processes. This paper describes an experimental design for the investigation of tumor vessel development and of therapeutic effects in a spatial and temporal manner. Using this setup, the stage of vessel development, tip cell and lumen formation, blood flow, extravasation, an established vascular bed, and vascular destruction can be visualized and followed. Furthermore, therapeutic effects, intratumoral fate, and the localization of chemotherapeutic compounds can also be followed.

This protocol describes the intravital imaging of transgenic mice expressing cell-specific fluorescent markers. Intravital imaging provides a non-invasive method for high-resolution observations in living animals using implantable windows through which the microscopic visualization of processes is possible. This method is especially useful to study longitudinal processes.

Videomicroscopia microscopia del sistema vascolare del tumore-associati utilizzando avanzate dorsale dello Skinfold finestra Chambers su topi transgenici fluorescenti

Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static ...

Using In Vitro Live-cell Imaging to Explore Chemotherapeutics Delivered by Lipid-based Nanoparticles

Conventional imaging techniques can provide detailed information about cellular processes. However, this information is based on static images in an otherwise dynamic system, and successive phases are easily overlooked or misinterpreted. Live-cell imaging and time-lapse microscopy, in which living cells can be followed for hours or even days in a more or less continuous fashion, are therefore very informative. The protocol described here allows for the investigation of the fate of chemotherapeutic nanoparticles after the delivery of doxorubicin (dox) in living cells. Dox is an intercalating agent that must be released from its nanocarrier to become biologically active. In spite of its clinical registration for more than two decades, its uptake, breakdown, and drug release are still not fully understood. This article explores the hypothesis that lipid-based nanoparticles are taken up by the tumor cells and are slowly degraded. Released dox is then translocated to the nucleus. To prevent fixation artifacts, live-cell imaging and time-lapse microscopy, described in this experimental procedure, can be applied.

Using In Vitro Live-cell Imaging to Explore Chemotherapeutics Delivered by Lipid-based Nanoparticles

Live-cell imaging is a powerful tool to visualize dynamic processes. The examination of fixed cells provides only static pictures, which can lead to misinterpretation and confusion about the process. This work presents a method to study uptake, drug release, and intracellular localization of liposomal nanoparticles in living cells.

Utilizzo In Vitro cellule vive di Imaging per esplorare chemioterapici consegnato da nanoparticelle di basata sui lipidi

Conventional imaging techniques can provide detailed information about cellular processes. However, this information is based on static images in an ...

Surgical Treatment for Benign Prostatic Hyperplasia: Holmium Laser Enucleation of the Prostate (HoLEP).

Benign prostatic hyperplasia (BPH) commonly occurs among elderly males. The condition causes symptoms in the lower urinary tract, thereby reducing the quality of life. BPH includes 2 major treatments: medication and surgery. Surgical treatment is often the most effective and final intervention. Holmium laser enucleation of the prostate (HoLEP), one of the most efficient surgical procedures for BPH, is conducted transurethrally. During surgery, the most important and most difficult part is to locate the surgical capsule of the prostate, which is consistent to the idea of anatomic surgery. Different skills may be needed in treating different parts of the prostate. HoLEP tends to be efficient and safe with good hemostatic properties, and able to treat bladder calculus, which may be the complications of BPH. The technique is particularly suitable for prostate of varying volumes and sizes. Indeed, HoLEP also presents certain disadvantages, such as a long learning curve and costly equipment. Regardless, this method may be the "new standard" and the excellent method for the surgical treatment of BPH.

Surgical Treatment for Benign Prostatic Hyperplasia: Holmium Laser Enucleation of the Prostate HoLEP.

Here we present a safe and efficient protocol, holmium laser enucleation of the prostate, to treat benign prostatic hyperplasia.

Trattamento chirurgico per l'iperplasia prostatica benigna: Holmium Laser Enucleation della prostata (HoLEP).

Benign prostatic hyperplasia (BPH) commonly occurs among elderly males. The condition causes symptoms in the lower urinary tract, thereby reducing the ...

Micro-dissection of Enamel Organ from Mandibular Incisor of Rats Exposed to Environmental Toxicants

Enamel defects resulting from environmental conditions and ways of life are public health concerns because of their high prevalence. These defects result from altered activity of cells responsible for enamel synthesis named ameloblasts, which present in enamel organ. During amelogenesis, ameloblasts follow a specific and precise sequence of events of proliferation, differentiation, and death. A rat continually growing incisors is a suitable experimental model to study ameloblast activity and differentiation stages in physiological and pathological conditions. Here, we describe a reliable and consistent method to micro-dissect enamel organ of rats exposed to environmental toxicants. The micro-dissected dental epithelia contain secretion- and maturation-stage ameloblasts that may be used for qualitative experiments, such as immunohistochemistry assays and in situ hybridization, as well as for quantitative analyses such as RT-qPCR, RNA-seq, and Western blotting.

Understanding enamel formation and possible alterations requires the study of ameloblast activity. Here, we describe a reliable and consistent method to micro-dissect enamel organs containing secretion- and maturation-stage ameloblasts that may be used for further quantitative and qualitative experimental procedures.

Micro-dissezione dell'organo dello smalto da incisivo dei ratti esposti a tossici ambientali

Enamel defects resulting from environmental conditions and ways of life are public health concerns because of their high prevalence. These defects result ...