The authors acknowledge the Carefusion Foundation for support via a clinical excellence grant of this work titled &#8220;Prospective evaluation of procedural povidone iodine rectal preparation to reduce infectious complications following ultrasound guided prostate needle biopsy.&#8221;

Este estudio relacionado paciente fue aprobado por la junta de revisión institucional (IRB) para la investigación humana. 1. Preparación para el paciente y Posicionamiento <ol><li> Llevar al paciente a la sala de procedimiento y colocarlos en su lado derecho o izquierdo con las rodillas flexionadas hacia el pecho. </li><li> Realice un tiempo de espera quirúrgica mediante el cual el paciente, procedimiento quirúrgico, y la indicación son revisados ​​y confirmados. </li><li> Cuando se indica, administrar sedación intravenosa a través del servicio de anestesia. </li></ol> 2. Realizar un examen rectal digital (DRE) <ol><li> Lugar tamaño guantes apropiados en ambas manos. </li><li> Inserte dedo enguantado después de la lubricación en la bóveda rectal y examinar la próstata para palpar posibles zonas sospechosas que necesitan atención enfocada durante la ecografía transrectal. 10 </li></ol> 3. Creación de povidona yodada Preparación rectal (PIRP) <ol><li> Verter 15 ml de una 10%solución de yodo povidona disponible en el mercado en un lebrillo estéril. </li><li> Añadir 5 ml de un gel de lidocaína 1% disponible comercialmente para esta cuenca. </li><li> Mezclar el contenido durante 15 s para crear una suspensión de povidona yodada. </li></ol> 4. Administración primaria de PIRP <ol><li> Use un hisopo rectal estéril para la cultura de la bóveda rectal. </li><li> Abre un 4 cm x 4 cm de gasa estéril, sumergirse en el lodo de yodo povidona, y luego insertar en la bóveda rectal. Sólo una pequeña cola de gasa se deja que emana del ano. </li><li> Deja la gasa en su lugar durante 2 min después de lo cual se retira y se desecha. </li></ol> 5. Administración Secundaria de PIRP <ol><li> Use un hisopo ginecológico desechable para pintar la zona perianal con la solución PIRP. </li><li> Inserte este hisopo ginecológica en la bóveda rectal a una distancia de 3 cm del ano. </li><li> Para maximizar aún más la exposición de la mucosa rectal para PIRP, limpiar la vau rectallt de una manera similar al menos tres veces. </li><li> Permitir que la solución de povidona yodada se seque durante 2 -. 3 min antes de proceder con la ecografía transrectal estándar y posterior biopsia 11 Esto es esencial ya que los efectos bactericidas de la PIRP requieren la solución a secar en la mucosa expuesta. </li><li> Use un hisopo rectal estéril para la cultura de la bóveda rectal. </li></ol> 6. La ecografía transrectal y biopsia <ol><li> Inserte la sonda de ultrasonido a través del recto. </li><li> Efectuar las mediciones de la próstata y la posterior biopsia de la manera estándar consistente con todas las prácticas urológicas. 11,12 </li></ol>

<ol>
	<li>Nam, R. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increasing+hospital+admission+rates+for+complications+after+transrectal+ultrasound+guided+prostate+biopsy.">Increasing hospital admission rates for complications after transrectal ultrasound guided prostate biopsy.</a> J Urol. 189, S12-S17 (2013).</li><li>Feliciano, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+incidence+of+fluoroquinolone+resistant+infections+after+prostate+biopsy+are+fluoroquinolones+still+effective+prophylaxis.">The incidence of fluoroquinolone resistant infections after prostate biopsy are fluoroquinolones still effective prophylaxis.</a> J Urol. 179 (3), 952-955 (2008).</li><li>Kehinde, O., Al-Maghrebi, M., Sheikh, M., Anim, J. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combined+ciprofloxacin+and+amikacin+prophylaxis+in+the+prevention+of+septicemia+after+transrectal+ultrasound+guided+biopsy+of+the+prostate.">Combined ciprofloxacin and amikacin prophylaxis in the prevention of septicemia after transrectal ultrasound guided biopsy of the prostate.</a> J Urol. 189 (3), 911-915 (2013).</li><li>Taylor, A. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+antimicrobial+prophylaxis+using+rectal+swab+cultures+in+men+undergoing+transrectal+ultrasound+guided+prostate+biopsy+is+associated+with+reduced+incidence+of+postoperative+infectious+complications+and+cost+of+care.">Targeted antimicrobial prophylaxis using rectal swab cultures in men undergoing transrectal ultrasound guided prostate biopsy is associated with reduced incidence of postoperative infectious complications and cost of care.</a> J Urol. 187 (4), 1275-1279 (2012).</li><li>Gyorfi, J. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peri+procedural+povidone+iodine+rectal+preparation+reduces+microorganism+counts+and+infectious+complications+following+ultrasound-guided+needle+biopsy+of+the+prostate.">Peri procedural povidone iodine rectal preparation reduces microorganism counts and infectious complications following ultrasound-guided needle biopsy of the prostate.</a> World J Urol. 32 (4), 905-909 (2014).</li><li>Park, D. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+infective+complications+of+transrectal+prostate+biopsy.">Control of infective complications of transrectal prostate biopsy.</a> Surg Infect Larchmt. 15 (4), 431-436 (2014).</li><li>Park, D. S., Oh, J. J., Lee, J. H., Jang, W. K., Hong, Y. K., Hong, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simple+use+of+the+suppository+type+povidone+iodine+can+prevent+infectious+complications+in+transrectal+ultrasound+guided+prostate+biopsy.">Simple use of the suppository type povidone iodine can prevent infectious complications in transrectal ultrasound guided prostate biopsy.</a> Adv Urol. , (2009).</li><li>AbuGhosh, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+prospective+randomized+trial+of+povidone+iodine+prophylactic+cleansing+of+the+rectum+before+transrectal+ultrasound+guided+prostate+biopsy.">A prospective randomized trial of povidone iodine prophylactic cleansing of the rectum before transrectal ultrasound guided prostate biopsy.</a> J Urol. 189 (4), 1326-1331 (2013).</li><li>Gerber, G. S., Brendler, C. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+the+Urological+Patient:+History,+Physical+Exam,+and+Urinalysis.">Evaluation of the Urological Patient: History, Physical Exam, and Urinalysis.</a> Campbell Walsh Urology. , (2015).</li><li>Hodge, K. K., McNeal, J. E., Terris, M. K., Stamey, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Random+systematic+versus+directed+ultrasound+guided+transrectal+core+biopsies+of+the+prostate.">Random systematic versus directed ultrasound guided transrectal core biopsies of the prostate.</a> J Urol. 142 (1), 71-74 (1989).</li><li>Donoghue, P. M., McSweeney, S. E., Jhaveri, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genitourinary+imaging:+current+and+emerging+applications.">Genitourinary imaging: current and emerging applications.</a> J Postgrad Med. 56 (2), 131-139 (2010).</li></ol>

The authors have nothing to disclose.

Infecciones siguientes ETR PNB son secundarios a la translocación de bacterias bóveda rectal en la próstata altamente vascular. Basándose en este principio, hemos explorado el simple método de utilización de un tema antiséptico tal como povidona yodada para reducir los recuentos de colonias de microorganismos antes de la biopsia con aguja de próstata. Nuestra experiencia pone de relieve una reducción del 4,3% al 0,6% (p = 0,02) con el uso de este régimen PIRP en el momento de la biopsia. Además, la hipótesis...

El cáncer de próstata es el cáncer más común en hombres y la segunda causa principal de mortalidad relacionada con el cáncer. En 2014, aproximadamente 233.000 casos serán diagnosticados en los Estados Unidos, con casi 30.000 hombres sucumbir a esta malignidad. 1 Si bien el antígeno prostático específico (PSA) en sangre y un examen rectal digital (DRE) desempeñan un papel esencial en la detección del cáncer de próstata, transrectal ecografía biopsia con aguja de próstata guiada (ETR PNB) es el medio más común para obtener un diagnóstico histológico del tejido. ETR PNB consiste en insertar una sonda de ultrasonido en el recto del paciente y luego generalmente aceptado 12-14 biopsias de la zona periférica de la próstata se encuentra justo delante. Cada biopsia con aguja requiere la aprobación de una aguja de 18 G a través de la pared del recto hasta la próstata altamente vascular. Por lo tanto, el riesgo teórico de sangrado e infección debido a la translocación bacteriana existe después del procedimiento. Ningunotheless, el post-procedimiento de tasa de complicaciones actuarial se ha mantenido históricamente bajo. La profilaxis antibiótica antes de la ETR PNB se prescribe habitualmente con antibióticos a base de quinolonas son los agentes más frecuentemente utilizados. A pesar de la profilaxis adecuada, la investigación contemporánea implica un aumento de las complicaciones infecciosas post-biopsia. 2 Además, los estudios han atribuido este aumento de las complicaciones infecciosas después de la ETR PNB a la aparición de microorganismos resistentes a quinolonas en particular E. coli. 3 Esta creciente tasa de resistencia a quinolonas sugiere la necesidad de regímenes de profilaxis alternativos para procedimientos ETR PNB. Con este fin, varias vías diferentes están bajo investigación. Un enfoque implica la administración de un antibiótico intravenosa o intramuscular en el momento de la biopsia en conjunción con una quinolona oral. 4 Si bien esta estrategia puede reducir los eventos siguientes sepsis TRUS PNB,una limitación principal de este enfoque es el potencial para desarrollar aún más los organismos resistentes a esta familia de antibióticos. Otra metodología recientemente investigado incorpora el uso de un hisopo rectal pre-biopsia para evaluar a los pacientes colonizados con flora rectal resistentes a las quinolonas. 5 Si se detectan tales organismos, un régimen antibiótico &quot;selectivo&quot; se podría emplear basado en el perfil de sensibilidad de las bacterias resistentes identificado cepa. Si bien la metodología es elegante, el proceso real de la obtención de este tipo de hisopos, selectivamente el cultivo en un medio selectivo quinolona, ​​y la adaptación de los antibióticos requiere a partir de entonces una infraestructura clínica y de laboratorio que pueden faltar en muchas de las prácticas clínicas. La administración de un tema antiséptica para reducir la bóveda rectal recuentos de colonias de microorganismos antes de la biopsia puede presentar una estrategia alternativa para limitar las infecciones TRUS PNB. Iodone povidona es un agente barato, fácilmente disponible que esse ha documentado que los recuentos bacterianos reducidos cuando se aplica a los sitios quirúrgicos. Aplicaciones en las dos cirugías ginecológicas y colorrectales son bien conocidos. Por lo tanto, el uso de yodo povidona como una preparación rectal presentaría un método producible, simple, y rentable a las infecciones TRUS PNB reducirse sin necesidad de preparación adicional antes de la biopsia. Con respecto a la literatura urológica, previamente trabajo ha observado una reducción en infecciones clínicas siguientes ETR PNB. 6.9 Este video muestra la técnica de PIRP para resaltar la facilidad y simplicidad de integrar en las prácticas clínicas. Todos los pacientes sometidos a ecografía transrectal PNB fueron vistos inicialmente en una clínica de urología de la especialidad para la remisión o evaluación inicial para la detección del cáncer de próstata. Esta evaluación implicó una revisión de antígeno del paciente prostático específico (PSA) y el rendimiento de un examen rectal digital (DRE). Anormalidad ya sea del PSA o DRE impulsó recomendaciónun PNB ETR. El procedimiento ETR PNB se discutió con todos los pacientes y los riesgos asociados, incluyendo infección, sangrado, y retención urinaria se aclararon. Todos los pacientes recibieron tres días de la profilaxis con antibióticos orales, ya sea con un medicamento a base de las quinolonas (por ejemplo, ciprofloxacina) o trimetoprim-sulfametoxazol (Bactrim) antes de la biopsia.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Povidone Iodine Solution, 10% Topical Microbicide Antiseptic, 8 fl oz.</td>
			<td>Equate</td>
			<td>4067979</td>
			<td></td>
		</tr>
		<tr>
			<td>4x4 sterile gauze pads, white</td>
			<td>Dukal Corporation</td>
			<td>DKL1412</td>
			<td></td>
		</tr>
		<tr>
			<td>disposable gynecologic swab&#160;</td>
			<td>numerous vendors</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1% lidocaine gel&#160;</td>
			<td>numerous vendors</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

povidone iodine rectal preparation at time of prostate needle biopsy is a simple and reproducible means to reduce risk of procedural infection

Una sola institución y los estudios basados ​​en la población destacan que las complicaciones infecciosas siguientes biopsia ecografía transrectal de próstata guiada aguja (ETR PNB) están aumentando. Estas infecciones son en gran parte atribuible a quinolonas microorganismos resistentes que colonizan la bóveda rectal y son trasladadas en el torrente sanguíneo durante el procedimiento de la biopsia. Una preparación rectal povidona yodada (PIRP) en el momento de la biopsia es un método simple, reproducible para reducir los recuentos de colonias de microorganismos rectales y por lo tanto las infecciones resultantes siguientes TRUS PNB. Todos los pacientes se administran tres días de tratamiento antibiótico oral antes de la biopsia. La técnica PIRP implica inicialmente la colocación del paciente en la forma estándar para una TRUS PNB. Tras el examen rectal digital, 15 ml de una solución al 10% de povidona yodada comercialmente disponible se mezcla con 5 ml de 1% de la jalea de lidocaína para crear suspensión. A 4 cm x 4 cm gasa estéril es soaked en esta suspensión y luego insertado en la bóveda rectal durante 2 min después de lo cual se retira. A partir de entonces, un hisopo de algodón ginecológico desechable se utiliza para pintar tanto en la zona perianal y la bóveda rectal a una distancia de 3 cm del ano. Después se deja que la solución de yodo povidona secar durante 2 - 3 min antes de proceder con la ecografía transrectal estándar y posterior biopsia. Esta técnica PIRP ha sido en la práctica en nuestra institución desde marzo de 2012, con una reducción asociada de las infecciones post-biopsia del 4,3% al 0,6% (p = 0,02). La principal ventaja de este régimen de profilaxis es su simplicidad y reproducibilidad con el uso de un agente fácilmente disponibles, de bajo costo para reducir las infecciones. Además, la técnica evita la exposición de los pacientes a los antibióticos sistémicos adicionales con potencial de propagación adicional de organismos resistentes a múltiples fármacos. El uso de PIRP en ETR PNB, sin embargo, no es aplicable para los pacientes con yodo o los mariscos unllergies.

Esta técnica PIRP ha sido en la práctica desde enero de 2012 (hasta el presente) con 165 pacientes que han sido inscritos en esta medida de mejora de la calidad. En este momento, sólo un paciente (0,6%) ha experimentado una complicación infecciosa en la forma de sepsis sistémica. Este paciente fue colonizado con una resistente E. múltiples fármacos coli cepa. Hisopos rectales tras la administración de PIRP en este paciente señalaron que la bóveda rectal todavía fue colonizado con bacterias 2...

Watch this Scientific Journal Video about Povidone Iodine Rectal Preparation at Time of Prostate Needle Biopsy is a Simple and Reproducible Means to Reduce Risk of Procedural Infection at JoVE.com

Povidone Iodine Rectal Preparation at Time of Prostate Needle Biopsy is a Simple and Reproducible Means to Reduce Risk of Procedural Infection

Here, we present a protocol for use at the time of transrectal ultrasound guided prostate needle biopsy (TRUS PNB) that is a simple and cost-effective means to reduce infections following the procedure.

Povidona yodada Preparación rectal al momento de la próstata Biopsia con aguja es un medio simple y reproducible para reducir el riesgo de infección Procesal

Single institution and population-based studies highlight that infectious complications following transrectal ultrasound guided prostate needle biopsy ...

povidone-iodine-rectal-preparation-at-time-prostate-needle-biopsy-is

Research

JoVE Journal

Medicine

31.8K vistas.  Penn State Milton S. Hershey Medical Center. Here, we present a protocol for use at the time of transrectal ultrasound guided prostate needle biopsy (TRUS PNB) that is a simple and cost-effective means to reduce infections following the procedure.

Video: Povidone Iodine Rectal Preparation at Time of Prostate Needle Biopsy is a Simple and Reproducible Means to Reduce Risk of Procedural Infection

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.

Evaluación de la absorción de nanopartículas en los tumores en tiempo real utilizando intravital imagen

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

Investigación

Medicina

MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR (qPCR)

MicroRNAs (miRNAs) are single-stranded, 18&#x2013;24 nucleotide long, non-coding RNA molecules. They are involved in virtually every cellular process including development1, apoptosis2, and cell cycle regulation3. MiRNAs are estimated to regulate the expression of 30% to 90% of human genes4 by binding to their target messenger RNAs (mRNAs)5. Widespread dysregulation of miRNAs has been reported in various diseases and cancer subtypes6. Due to their prevalence and unique structure, these small molecules are likely to be the next generation of biomarkers, therapeutic agents and/or targets.
Methods used to investigate miRNA expression include SYBR green I dye- based as well as Taqman-probe based qPCR. If miRNAs are to be effectively used in the clinical setting, it is imperative that their detection in fresh and/or archived clinical samples be accurate, reproducible, and specific. qPCR has been widely used for validating expression of miRNAs in whole genome analyses such as microarray studies7. The samples used in this protocol were from patients who underwent radical prostatectomy for clinically localized prostate cancer; however other tissues and cell lines can be substituted in. Prostate specimens were snap-frozen in liquid nitrogen after resection. Clinical variables and follow-up information for each patient were collected for subsequent analysis8.
Quantification of miRNA levels in prostate tumor samples. The main steps in qPCR analysis of tumors are: Total RNA extraction, cDNA synthesis, and detection of qPCR products using miRNA-specific primers. Total RNA, which includes mRNA, miRNA, and other small RNAs were extracted from specimens using TRIzol reagent. Qiagen's miScript System was used to synthesize cDNA and perform qPCR (Figure 1). Endogenous miRNAs are not polyadenylated, therefore during the reverse transcription process, a poly(A) polymerase polyadenylates the miRNA. The miRNA is used as a template to synthesize cDNA using oligo-dT and Reverse Transcriptase. A universal tag sequence on the 5' end of oligo-dT primers facilitates the amplification of cDNA in the PCR step. PCR product amplification is detected by the level of fluorescence emitted by SYBR Green, a dye which intercalates into double stranded DNA. Specific miRNA primers, along with a Universal Primer that binds to the universal tag sequence will amplify specific miRNA sequences. 
The miScript Primer Assays are available for over a thousand human-specific miRNAs, and hundreds of murine-specific miRNAs. Relative quantification method was used here to quantify the expression of miRNAs. To correct for variability amongst different samples, expression levels of a target miRNA is normalized to the expression levels of a reference gene. The choice of a gene on which to normalize the expression of targets is critical in relative quantification method of analysis. Examples of reference genes typically used in this capacity are the small RNAs RNU6B, RNU44, and RNU48 as they are considered to be stably expressed across most samples. In this protocol, RNU6B is used as the reference gene.

MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR qPCR

Quantitative Real Time polymerase chain reaction (qPCR) is a rapid and sensitive method to investigate the expression levels of various microRNA (miRNA) molecules in tumor samples. Using this method expression of hundreds of different miRNA molecules can be amplified, quantified, and analyzed from the same cDNA template.

Detección de microARN en tumores de próstata por el cuantitativo PCR en tiempo real (qPCR)

MicroRNAs (miRNAs) are single-stranded, 18&#x2013;24 nucleotide long, non-coding RNA molecules. They are involved in virtually every cellular process ...

Combined In vivo Optical and &#181;CT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

Multimodality imaging has emerged as a common technological approach used in both preclinical and clinical research. Advanced techniques that combine in vivo optical and &#956;CT imaging allow the visualization of biological phenomena in an anatomical context. These imaging modalities may be especially useful to study conditions that impact bone. In particular, orthopaedic implant infections are an important problem in clinical orthopaedic surgery. These infections are difficult to treat because bacterial biofilms form on the foreign surgically implanted materials, leading to persistent inflammation, osteomyelitis and eventual osteolysis of the bone surrounding the implant, which ultimately results in implant loosening and failure. Here, a mouse model of an infected orthopaedic prosthetic implant was used that involved the surgical placement of a Kirschner-wire implant into an intramedullary canal in the femur in such a way that the end of the implant extended into the knee joint. In this model, LysEGFP mice, a mouse strain that has EGFP-fluorescent neutrophils, were employed in conjunction with a bioluminescent Staphylococcus aureus strain, which naturally emits light. The bacteria were inoculated into the knee joints of the mice prior to closing the surgical site. In vivo bioluminescent and fluorescent imaging was used to quantify the bacterial burden and neutrophil inflammatory response, respectively. In addition, &#956;CT imaging was performed on the same mice so that the 3D location of the bioluminescent and fluorescent optical signals could be co-registered with the anatomical &#956;CT images. To quantify the changes in the bone over time, the outer bone volume of the distal femurs were measured at specific time points using a semi-automated contour based segmentation process. Taken together, the combination of in vivo bioluminescent/fluorescent imaging with &#956;CT imaging may be especially useful for the noninvasive monitoring of the infection, inflammatory response and anatomical changes in bone over time.

Combined In vivo Optical and  &amp;#181;CT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

Combined optical and &#956;CT imaging in a mouse model of orthopaedic implant infection, utilizing a bioluminescent engineered strain of Staphylococcus aureus, provided the capability to noninvasively and longitudinally monitor the dynamics of the bacterial infection, as well as the corresponding inflammatory response and anatomical changes in the bone.

Combinado<em&gt; En vivo</em&gt; Óptica y μCT imágenes para vigilar la infección, inflamación y Hueso anatomía en una infección de implantes ortopédicos en ratones

Multimodality imaging has emerged as a common technological approach used in both preclinical and clinical research. Advanced techniques that combine in ...

Substernal Thyroid Biopsy Using Endobronchial Ultrasound-guided Transbronchial Needle Aspiration

Substernal thyroid goiter (STG) represents about 5.8% of all mediastinal lesions1. There is a wide variation in the published incidence rates due to the lack of a standardized definition for STG. Biopsy is often required to differentiate benign from malignant lesions. Unlike cervical thyroid, the overlying sternum precludes ultrasound-guided percutaneous fine needle aspiration of STG. Consequently, surgical mediastinoscopy is performed in the majority of cases, causing significant procedure related morbidity and cost to healthcare. Endobronchial Ultrasound-guided Transbronchial Needle Aspiration (EBUS-TBNA) is a frequently used procedure for diagnosis and staging of non-small cell lung cancer (NSCLC). Minimally invasive needle biopsy for lesions adjacent to the airways can be performed under real-time ultrasound guidance using EBUS. Its safety and efficacy is well established with over 90% sensitivity and specificity. The ability to perform EBUS as an outpatient procedure with same-day discharges offers distinct morbidity and financial advantages over surgery. As physicians performing EBUS gained procedural expertise, they have attempted to diversify its role in the diagnosis of non-lymph node thoracic pathologies. We propose here a role for EBUS-TBNA in the diagnosis of substernal thyroid lesions, along with a step-by-step protocol for the procedure.

Substernal thyroid lesions are common, and need to be differentiated from malignancy. Obtaining percutaneous fine needle biopsy is not possible due to its retrosternal location. This article proposes a protocol for biopsy of substernal thyroid lesions using Endobronchial Ultrasound-guided Transbronchial Needle Aspiration (EBUS-TBNA).

Subesternal tiroides Biopsia Usando guiada por ecografía endobronquial punción transbronquial

Substernal thyroid goiter (STG) represents about 5.8% of all mediastinal lesions1. There is a wide variation in the published incidence rates due to the ...

Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining samples of human Vastus lateralis, isolating mitochondria from minimal amounts of skeletal muscle tissue, and plate based respirometric profiling using an extracellular flux (XF) analyzer. Comparison of respirometric profiles obtained using 1.0, 2.5 and 5.0 &#956;g of mitochondria indicate that 1.0 &#956;g is sufficient to measure respiration and that 5.0 &#956;g provides most consistent results based on comparison of standard errors. Western blot analysis of isolated mitochondria for mitochondrial marker COX IV and non-mitochondrial tissue marker GAPDH indicate that there is limited non-mitochondrial contamination using this protocol. The ability to study mitochondrial respirometry in as little as 20 mg of muscle tissue allows users to utilize individual biopsies for multiple study endpoints in clinical research projects.

Methods for biopsy of Vastus lateralis, preparation of purified mitochondria, and respirometric profiling are described. The use of small muscle volume makes this technique suitable for clinical research applications.

Evaluación de Preparación y respirometría de mitocondrias aisladas de tejido músculo esquelético Obtenidos percutánea Biopsia con aguja

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining ...

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

A-crítico tamaño femoral Defecto sencillo modelo en ratones

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe the use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations of superior and uniform quality to that which can be achieved by hand. The device comprises a base that holds 4 stainless steel rods and a top, which acts a cutting guide. The rods are inserted into the lumen of the small intestine [divided into thirds] and the colon. The rods and samples are then placed over a piece of filter paper or card into the holding slots in the base of the device. The top of the device is then positioned and serves as a cutting guide. The two angled sections in the center of the top piece are used to guide a knife or scalpel and cut the intestines longitudinally on the top of the rods. Once the intestinal sections have been cut, the top is removed and the card,&#160;tissue and rods gently removed from the device and placed on the bench. The rods are then gently rolled sideways to flatten and stick the intestinal segments onto the underlying piece of filter paper or card. The final preparation can then be examined or fixed and stored for later analysis. The preparations are invaluable for the study of intestinal changes in normal or genetically modified mouse models. The preparations have been used for the study and quantification of the effects of inflammation (colitis), damage, pre-cancerous lesions (aberrant crypt foci (ACFs) and mucin depleted foci (MDFs)) and polyps or tumors.

The use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations is described.

Un dispositivo sencillo de preparar rápidamente enteros Montes del Intestino Ratón

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe ...

Generation of Prostate Cancer Patient Derived Xenograft Models from Circulating Tumor Cells

Patient derived xenograft (PDX) models are gaining popularity in cancer research and are used for preclinical drug evaluation, biomarker identification, biologic studies, and personalized medicine strategies. Circulating tumor cells (CTC) play a critical role in tumor metastasis and have been isolated from patients with several tumor types. Recently, CTCs have been used to generate PDX experimental models of breast and prostate cancer. This manuscript details the method for the generation of prostate cancer PDX models from CTCs developed by our group. Advantages of this method over conventional PDX models include independence from surgical sample collection and generating experimental models at various disease stages. Density gradient centrifugation followed by red blood cell lysis and flow cytometry depletion of CD45 positive mononuclear cells is used to enrich CTCs from peripheral blood samples collected from patients with metastatic disease. The CTCs are then injected into immunocompromised mice; subsequently generated xenografts can be used for functional studies or harvested for molecular characterization. The primary limitation of this method is the negative selection method used for CTC enrichment. Despite this limitation, the generation of PDX models from CTCs provides a novel experimental model to be applied to prostate cancer research.

This manuscript details a method used to generate prostate cancer patient derived xenografts (PDXs) from circulating tumor cells (CTCs). The generation of PDX models from CTCs provides an alternative experimental model to study prostate cancer; the most commonly diagnosed tumor and a frequent cause of death from cancer in men.

Generación del paciente con cáncer de próstata Derivado modelos de xenoinjertos de células tumorales circulantes

Patient derived xenograft (PDX) models are gaining popularity in cancer research and are used for preclinical drug evaluation, biomarker identification, ...

A Simple Mechanical Procedure to Create Limbal Stem Cell Deficiency in Mouse

Limbal stem cell deficiency (LSCD) is a state of malfunction or loss of limbal epithelial stem cells, after which the corneal epithelium is replaced with conjunctiva. Patients suffer from recurrent corneal defects, pain, inflammation, and loss of vision.
Previously, a murine model of LSCD was described and compared to two other models. The goal was to produce a consistent mouse model of LSCD that both mimics the phenotype in humans and lasts long enough to make it possible to study the disease pathophysiology and to evaluate new treatments. Here, the technique is described in more detail.
A motorized tool with a rotating burr has been designed to remove the rust rings from the corneal surface or to smooth the pterygium bed in patients. It is a suitable device to create the desired LSCD model. It is a readily available, easy-to-use tool with a fine tip that makes it appropriate for working on small eyes, as in mice. Its application prevents unnecessary trauma to the eye and it does not result in unwanted injuries, as often is the case with chemical injury models. As opposed to a blunt scraper, it removes the epithelium with the basement membrane. In this protocol, the limbal area was abraded two times, and then the whole corneal epithelium was shaved from limbus to limbus. To avoid stroma injury, care was taken not to brush the corneal surface once the epithelium was already removed.

The following article provides an easy, reproducible technique to effectively create a sustainable mouse model of limbal stem cell deficiency (LSCD). This animal model is useful in testing and comparing the efficacy of treatments for limbal stem cell diseases.

Un procedimiento mecánico simple de crear deficiencia de células madre limbares en ratón

Limbal stem cell deficiency (LSCD) is a state of malfunction or loss of limbal epithelial stem cells, after which the corneal epithelium is replaced with ...

An Anatomical Study of Nerves at Risk During Minimally Invasive Hallux Valgus Surgery

The growing popularity of minimally invasive surgical (MIS) procedures makes it necessary that new anatomical references arise, to aid in tridimensional orientation and localization of structures that are not directly visible to the surgeon. This is especially critical for structures at risk like nerves or blood vessels. Optimization of the handling of cadaveric material and the combination of multiple techniques compensate for the limited availability of adequate specimens. The described protocol combines anatomical plane-by-plane dissection and sectional anatomy of fresh-frozen specimens to help localize relevant structures, such as nerves, arteries, veins and to correctly position the portals during MIS procedures. Depiction of these structures in anatomy textbooks can differ from what is encountered in the surgical field; and for this reason, new anatomical studies with a surgical orientation are needed. However, this is a complex, time-consuming technique requiring specific training. The anatomical references described with the so-called 'clock method' provide the surgeon with an easy and reproducible system to locate the path of the nerves at risk in Hallux Valgus MIS procedures. This model can be extrapolated to many other minimally invasive surgical procedures.

Minimally invasive surgical (MIS) procedures rely on anatomical references to localize structures not directly visible to the surgeon. This manuscript describes a combined method of plane-by-plane dissection and sectional anatomy of fresh-frozen specimens to locate the structures at risk during MIS procedures.

Un estudio anatómico de los nervios en riesgo durante la cirugía de mínimamente invasiva de Hallux Valgus

The growing popularity of minimally invasive surgical (MIS) procedures makes it necessary that new anatomical references arise, to aid in tridimensional ...