מחקר זה נתמך על ידי מענקי NIH R37 DK54824 וR01 DK57284 לק&quot;ג

החוקרים מצהירים כי אין להם אינטרסים כלכליים מתחרים.

במאמר זה אנו מתארים פשוטים בשיטת מבחנה שריר חלק התכווצות שיכול לשמש כדי לטפל במספר השאלות מדעיות חשובות הקשורים לפיזיולוגיה בשלפוחית ​​השתן ופתולוגיה, כמו גם סיוע לגילוי תרופות חדשות לטיפול בתפקוד שלפוחית ​​השתן. יש לנו הדגמתי את השימוש בשיטה זו להערכת מאפי?...

שריר החלק בשלפוחית ​​השתן נרפה כדי לאפשר אחסון שתן, וחוזים לעורר חיסול שתן. הרפיה מתווכת על ידי תכונות שריר חלק פנימיות ועל ידי שחרור טוניק של נוראפינפרין (NE) מהעצבים הסימפתטית, אשר מפעיל קולטני בטא adrenergic (β 3 AR באדם) בdetrusor. רקה מושגת על ידי עיכוב הקלט האוהד והפעלת העצבים הפאראסימפתטית שישחררו ACH / ATP להתכווץ שריר החלק בשלפוחית ​​השתן 1. תנאים רבים פתולוגיים, כולל מוח ו / או פגיעה בחוט השדרה, מחלות ניווניות, סוכרת, חסימת מוצא שלפוחית ​​השתן או דלקת שלפוחית ​​השתן interstitial, יכולים לשנות את תפקוד שלפוחית ​​השתן עמוק, עם השלכות קשות על איכות חיים של המטופל 2. תנאים אלה לשנות את ההתכווצות של שריר החלק על ידי המשפיע על אחד או יותר מרכיבים של שלפוחית ​​השתן: שריר החלק, מביא או עצבי efferent ו / אורירית. כמה in vivo ובשיטות מבחנה ללמוד תפקוד שלפוחית ​​השתן פותחו. בvivo, cystometry הוא המדידה הראשונית של תפקוד שלפוחית ​​השתן. למרות זאת היא הכנה שלמה המאפשרת איסוף מידע במסגרת קרובה לתנאים פיסיולוגיים, יש מספר הנסיבות שבהן השימוש ברצועות שריר חלק הוא מועדפת. אלה כוללים מצבים שבם וכירורגי / או מניפולציות תרופתי ישפיעו על ההישרדות והיציבות של הכנת in vivo, או כאשר המחקרים דורשים שימוש ברקמה האנושית או כימיקלים יקרים. שיטה זו גם מאפשרת בדיקה של ההשפעות של תרופות, גיל ופתולוגיה בכל רכיב של שלפוחית ​​השתן, כלומר שריר חלק, רירית, מביא ועצבי efferent. רצועות שלפוחית ​​כבר מועסקות במשך השנים על ידי קבוצות רבות לענות על מספר שאלות מדעיות. הם היו רגילים לevaשינויי luate בפעילות ספונטנית myogenic הנגרמים על ידי פתולוגיה. הוא האמין כי פעילות זו תורמת לתסמיני דחיפות ותכיפות של פעילות יתר של שלפוחית ​​(OAB), ולכן הוא יעד לתרופות שפותחו עבור OAB 3-9. רצועות שלפוחית ​​השתן שמשו גם כדי לחקור גורמי myogenic ועצביים המווסתים את טונוס שרירים חלק במטרה לגלות תעלות יונים ו / או קולטנים ו / או מסלולים תאיים שיכול להיות ממוקדת כדי לגרום או הרפיה או התכווצות של שריר חלק 3,10- 13. מחקרים אחרים התמקדו בטבע של עצבית, כולל משדרים וקולטנים המעורבים ושינויים הנגרמים על ידי פתולוגיה 14,15. בנוסף, השיטה הייתה בשימוש להשוואה בין רקמות ממינים שונים 16- 18, בין האיברים 19-21, והערכה של מערכות יחסים מבנה פעילות תרופת 22-24. סיומת של שיטה זו נעשתה שימוש כדי measurדואר את ההשפעה של תרופות על שחרור משדר מעצבי efferent 25. יתר על כן, מגוון רחב של רקמות (שלפוחית ​​השתן, שופכה, מערכת עיכול, במערכת העיכול) שנקטפו מבעלי חיים או בני אדם (מניתוחים או רקמת תורם איברים שאושרו למחקר) וממגוון רחב של מודלים של בעלי חיים, כולל פגיעה בחוט השדרה (SCI), חסימת מוצא שלפוחית ​​השתן (BOO), או דלקת שלפוחית ​​השתן interstitial (IC) יכול להיחקר שימוש בטכניקה זו. במאמר זה אנו מציגים את השימוש בשיטה זו יחד עם פרוטוקולי ניסוי הכרחיים, כדי לטפל בכמה שאלות מדעיות שהוזכרו לעיל.

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bladder smooth muscle strip contractility as a method to evaluate lower urinary tract pharmacology

אנו מתארים שיטה במבחנה למדידת התכווצות שריר חלק בשלפוחית ​​השתן, ושימוש בו לחקירת מאפיינים פיסיולוגיים ותרופתיים של שריר החלק, כמו גם שינויים הנגרמים על ידי פתולוגיה. שיטה זו מספקת מידע קריטי להבנת תפקוד שלפוחית ​​השתן תוך כדי התגברות על קשיים גדולים מתודולוגיים נתקלו בניסויי vivo, כגון מניפולציות כירורגית ותרופתיות המשפיעות על יציבות והישרדות של ההכנות, שימוש ברקמות אנושיות, ו / או שימוש בכימיקלים יקרים. הוא גם מספק דרך לחקור את המאפיינים של כל מרכיב בשלפוחית ​​השתן (כלומר שריר חלק, רירית, בעצבים) בתנאים בריאים ופתולוגי. שלפוחית ​​השתן הוא להסיר חיה הרדים, להציב פתרון קרבס וחתוך לרצועות. רצועות ממוקמות לתוך תא מלא בפתרון קרבס חם. קצה אחד מחובר לtensio איזומטרימתמר n למדוד כוח התכווצות, בקצה השני מחובר למוט קבוע. רקמה היא מגורה על ידי ישירות הוספת תרכובות באמבטיה או על ידי אלקטרודות גירוי שדה חשמליות המפעילות את עצבים, בדומה למפעיל התכווצויות שלפוחית ​​in vivo. אנו מדגימים את השימוש בשיטה זו על מנת להעריך התכווצות שריר חלק ספונטנית במהלך פיתוח, ולאחר פגיעה בחוט השדרה ניסיונית, הטבע של עצבית (משדרים וקולטנים מעורבים), גורמים מעורבים בויסות של פעילות שריר חלק, התפקיד של רכיבי שלפוחית ​​בודדים, ומינים והבדלי איבר בתגובה לסוכנים תרופתיים. בנוסף, הוא יכול לשמש לחקר מסלולים תאיים מעורבים בהתכווצות ו / או הרפיה של שריר החלק, יחסי מבנה פעילות תרופה והערכה של שחרור משדר. שיטת התכווצות שריר חלק במבחנה כבר נעשה שימוש נרחב FOr מעל 50 שנים, וספק נתונים שתרמו באופן משמעותי להבנה של תפקוד שלפוחית ​​שלנו, כמו גם לפיתוח תרופות של תרכובות המשמשות כיום מבחינה קלינית לניהול שלפוחית ​​השתן.

פעילות myogenic ספונטנית פעילות myogenic ספונטנית היא אופייני שריר חלק חשוב שעובר שינויים עם התפתחות לאחר הלידה 6-9 ופתולוגיה (למשל, SCI, BOO) 3-5. משום שהאמינה כי פעילות זו תורמת לסימפטומים של פעילות יתר של שלפוחית ​?...

Watch this Scientific Journal Video about Bladder Smooth Muscle Strip Contractility as a Method to Evaluate Lower Urinary Tract Pharmacology at JoVE.com

Bladder Smooth Muscle Strip Contractility as a Method to Evaluate Lower Urinary Tract Pharmacology

כתב יד זה מציג פשוט, אך רב עוצמה, בשיטה חוץ גופית להערכת התכווצות שריר חלק בתגובה לסוכנים תרופתיים או גירוי עצב. יישומים עיקריים הם הקרנת סמים ופיזיולוגיה רקמת הבנה, פרמקולוגיה, ופתולוגיה.

שריר חלק בשלפוחית ​​השתן רצועת התכווצות כשיטה להערכת נמוכה יותר בדרכי שתן פרמקולוגיה

We describe an in vitro method to measure bladder smooth muscle contractility, and its use for investigating physiological and pharmacological properties ...

bladder-smooth-muscle-strip-contractility-as-method-to-evaluate-lower

Research

JoVE Journal

Medicine

26.1K Views.  University of Pittsburgh School of Medicine.  כתב יד זה מציג פשוט, אך רב עוצמה, בשיטה חוץ גופית להערכת התכווצות שריר חלק בתגובה לסוכנים תרופתיים או גירוי עצב. יישומים עיקריים הם הקרנת סמים ופיזיולוגיה רקמת הבנה, פרמקולוגיה, ופתולוגיה. 

Video: Bladder Smooth Muscle Strip Contractility as a Method to Evaluate Lower Urinary Tract Pharmacology

Manual Muscle Testing:  A Method of Measuring Extremity Muscle Strength Applied to Critically Ill Patients

Survivors of acute respiratory distress syndrome (ARDS) and other causes of critical illness often have generalized weakness, reduced exercise tolerance, and persistent nerve and muscle impairments after hospital discharge.1-6 Using an explicit protocol with a structured approach to training and quality assurance of research staff, manual muscle testing (MMT) is a highly reliable method for assessing strength, using a standardized clinical examination, for patients following ARDS, and can be completed with mechanically ventilated patients who can tolerate sitting upright in bed and are able to follow two-step commands. 7, 8 
This video demonstrates a protocol for MMT, which has been taught to &ge;43 research staff who have performed &gt;800 assessments on &gt;280 ARDS survivors. Modifications for the bedridden patient are included. Each muscle is tested with specific techniques for positioning, stabilization, resistance, and palpation for each score of the 6-point ordinal Medical Research Council scale.7,9-11 Three upper and three lower extremity muscles are graded in this protocol: shoulder abduction, elbow flexion, wrist extension, hip flexion, knee extension, and ankle dorsiflexion. These muscles were chosen based on the standard approach for evaluating patients for ICU-acquired weakness used in prior publications. 1,2.

Survivors of acute respiratory distress syndrome (ARDS) and critical illness frequently develop long-lasting muscle weakness. Manual muscle testing (MMT) is a standardized clinical examination commonly used to measure strength of peripheral skeletal muscle groups. This video demonstrates MMT using the 6-point Medical Research Council scale.

בדיקת שרירים ידנית: שיטת מדידת חוזק שרירים בגפיים יישומית על חולים אנושים

Survivors of acute respiratory distress syndrome (ARDS) and other causes of critical illness often have generalized weakness, reduced exercise tolerance, ...

Probe-based Confocal Laser Endomicroscopy of the Urinary Tract: The Technique

Probe-based confocal laser endomicroscopy (CLE) is an emerging optical imaging technology that enables real-time in vivo microscopy of mucosal surfaces during standard endoscopy. With applications currently in the respiratory1 and gastrointestinal tracts,2-6 CLE has also been explored in the urinary tract for bladder cancer diagnosis.7-10 Cellular morphology and tissue microarchitecture can be resolved with micron scale resolution in real time, in addition to dynamic imaging of the normal and pathological vasculature.7
The probe-based CLE system (Cellvizio, Mauna Kea Technologies, France) consists of a reusable fiberoptic imaging probe coupled to a 488 nm laser scanning unit. The imaging probe is inserted in the working channels of standard flexible and rigid endoscopes. An endoscope-based CLE system (Optiscan, Australia), in which the confocal endomicroscopy functionality is integrated onto the endoscope, is also used in the gastrointestinal tract. Given the larger scope diameter, however, application in the urinary tract is currently limited to ex vivo use.11 Confocal image acquisition is done through direct contact of the imaging probe with the target tissue and recorded as video sequences. As in the gastrointestinal tract, endomicroscopy of the urinary tract requires an exogenenous contrast agent&mdash;most commonly fluorescein, which can be administered intravenously or intravesically. Intravesical administration is a well-established method to introduce pharmacological agents locally with minimal systemic toxicity that is unique to the urinary tract. Fluorescein rapidly stains the extracellular matrix and has an established safety profile.12 Imaging probes of various diameters enable compatibility with different caliber endoscopes. To date, 1.4 and 2.6 mm probes have been evaluated with flexible and rigid cystoscopy.10 Recent availability of a &lt; 1 mm imaging probe13 opens up the possibility of CLE in the upper urinary tract during ureteroscopy. Fluorescence cystoscopy (i.e. photodynamic diagnosis) and narrow band imaging are additional endoscope-based optical imaging modalities14 that can be combined with CLE to achieve multimodal imaging of the urinary tract. In the future, CLE may be coupled with molecular contrast agents such as fluorescently labeled peptides15 and antibodies for endoscopic imaging of disease processes with molecular specificity.

Probe-based confocal laser endomicroscopy enables real-time microscopy of the human urinary tract during cystoscopy, providing dynamic, intravital imaging of pathological states such as bladder cancer with cellular resolution. Endomicroscopy may augment the diagnostic accuracy of standard white light endoscopy and provide intraoperative image guidance to improve surgical resection.

Endomicroscopy בדיקה מבוסס Confocal ליזר בדרכי השתן: הטכניקה

Probe-based  confocal laser endomicroscopy (CLE) is an emerging optical imaging technology  that enables real-time in vivo microscopy of mucosal surfaces ...

A Modified Precipitation Method to Isolate Urinary Exosomes

Identification of biomarkers that allow early detection of kidney diseases in urine and plasma has been an area of active interest for several years. Urinary exosome vesicles, 40-100 nm in size, are released into the urine under normal conditions by cells from all nephron segments and may contain protein, mRNA and microRNA representative of their cell type of origin. Under conditions of renal dysfunction or injury, exosomes may contain altered proportions of these components, which may serve as biomarkers for disease. There are currently several methods available for isolation of urinary exosomes, and we have previously conducted an experimental comparison of each of these approaches, including three based on ultracentrifugation, one using a nanomembrane ultrafiltration concentrator, one using a commercial precipitation reagent and one using a modification of the precipitation technique using ExoQuick reagent that we developed in our laboratory. We found the modified precipitation method produced the highest yield of exosome particles, miRNA, and mRNA, making this approach suitable for the isolation of exosomes for subsequent RNA profiling. We conclude that the modified exosome precipitation method offers a quick, scalable, and effective alternative for the isolation of exosomes from urine. In this report, we describe our modified precipitation technique using ExoQuick reagent for isolating exosomes from human urine.

This manuscript describes a protocol for isolating exosomes from human urine using a modified precipitation technique.

שיטת ממטרים השתנה לבודד השתן Exosomes

Identification of biomarkers that allow early detection of kidney diseases in urine and plasma has been an area of active interest for several years. ...

Urinary Bladder Distention Evoked Visceromotor Responses as a Model for Bladder Pain in Mice

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized by increased urgency and frequency of urination, as well as nocturia and general pelvic pain, especially upon bladder filling or voiding. Despite years of research, the cause of IC/BPS remains elusive and treatment strategies are unable to provide complete relief to patients. In order to study nervous system contributions to the condition, many animal models have been developed to mimic the pain and symptoms associated with IC/BPS. One such murine model is urinary bladder distension (UBD). In this model, compressed air of a specific pressure is delivered to the bladder of a lightly anesthetized animal over a set period of time. Throughout the procedure, wires in the superior oblique abdominal muscles record electrical activity from the muscle. This activity is known as the visceromotor response (VMR) and is a reliable and reproducible measure of nociception. Here, we describe the steps necessary to perform this technique in mice including surgical manipulations, physiological recording, and data analysis. With the use of this model, the coordination between primary sensory neurons, spinal cord secondary afferents, and higher central nervous system areas involved in bladder pain can be unraveled. This basic science knowledge can then be clinically translated to treat patients suffering from IC/BPS.

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized in part by pelvic pain. In order to study nervous system contributions to the condition, a physiological model of urinary bladder pain is used in mice and rats.

תגובות שתן בשלפוחית ​​השתן ההתנפחות העוררת Visceromotor כמודל לשלפוחית ​​שתן כאב בעכברים

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition ...

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

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

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

הסתיידות של לתאי השריר החלק בכלי הדם והדמיה של אבי העורקים הסתיידות ודלקת

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

Contractility Measurements of Human Uterine Smooth Muscle to Aid Drug Development

Discovery and characterization of novel pharmaceutical compounds or biochemical probes rely on robust and physiologically relevant assay systems. We describe methods to measure ex vivo myometrium contractility. This assay can be used to investigate factors and molecules involved in the modulation of myometrial contraction and to determine their excitatory or inhibitory actions, and hence their therapeutic potential in vivo. Biopsies are obtained from women undergoing cesarean section delivery with informed consent. Fine strips of myometrium are dissected, clipped and attached to a force transducer within 1 mL organ baths superfused with physiological saline solution at 37 &#176;C. Strips develop spontaneous contractions within 2&#8211;3 h under set tension and remain stable for many hours (&#62;6 h). Strips can also be stimulated to contract such as by the endogenous hormones, oxytocin and vasopressin, which cause concentration-dependent modulation of contraction frequency, force and duration, to more closely resemble contractions in labor. Hence, the effect of known and novel drug leads can be tested on spontaneous and agonist-induced contractions.
This protocol specifically details how this assay can be used to determine the potency of known and novel agents by measuring their effects on various parameters of human myometrial contraction. We use the oxytocin- and V1a receptor antagonists, atosiban and SR49059 as examples of known compounds which inhibit oxytocin- and vasopressin-induced contractions, and demonstrate how this method can be used to complement and validate pharmacological data obtained from cell-based assays to aid drug development. The effects of novel agonists in comparison to oxytocin and vasopressin can also be characterized. Whilst we use the example of the oxytocin/ vasopressin system, this method can also be used to study other receptors and ion channels that play a role in uterine contraction and relaxation to advance the understanding of human uterine physiology and pathophysiology.

This article describes experimental protocols to study ex vivo contractions of human myometrium and their application in drug discovery. This technique is used to improve the understanding of myometrial physiology and pathophysiology as well as to validate pharmacological data from novel research probes or drug leads.

Contractility המידות של האדם הרחם שריר חלק לפיתוח התרופה עזרה

Discovery and characterization of novel pharmaceutical compounds or biochemical probes rely on robust and physiologically relevant assay systems. We ...

Microelectrode Impalement Method to Record Membrane Potential from a Cannulated Middle Cerebral Artery

Membrane potential (Vm) of vascular smooth muscle cells determines vessel tone and thus blood flow to an organ. Changes in the expression and function of ion channels and electrogenic pumps that regulate Vm in disease conditions could potentially alter Vm, vascular tone, and blood flow. Thus, a basic understanding of electrophysiology and the methods necessary to accurately record Vm in healthy and diseased states are essential. This method will allow modulating Vm using different pharmacological agents to restore Vm. Although there are several methods, each with its advantages and disadvantages, this article provides protocols to record Vm from cannulated resistance vessels such as the middle cerebral artery using the microelectrode impalement method. Middle cerebral arteries are allowed to gain myogenic tone in a myograph chamber, and the vessel wall is impaled using high resistance microelectrodes. The Vm signal is collected through an electrometer, digitized, and analyzed. This method provides an accurate reading of the Vm of a vessel wall without damaging the cells and without changing the membrane resistance.

The primary goal of this article is to provide details of how to record membrane potential (Vm) from the middle cerebral artery using the microelectrode impalement method. The cannulated middle cerebral artery is equilibrated to gain myogenic tone, and the vessel wall is impaled using high resistance microelectrodes.

שיטת החלוקה של מיקרו-אלקטרודה להקלטת פוטנציאל ממברנה מעורק מוחי אמצעי של קאננולוב

Membrane potential (Vm) of vascular smooth muscle cells determines vessel tone and thus blood flow to an organ. Changes in the expression and function of ...

Quantitative Analysis of Cellular Composition in Advanced Atherosclerotic Lesions of Smooth Muscle Cell Lineage-Tracing Mice

Atherosclerosis remains the leading cause of death worldwide and, despite countless preclinical studies describing promising therapeutic targets, novel interventions have remained elusive. This is likely due, in part, to a reliance on preclinical prevention models investigating the effects of genetic manipulations or pharmacological treatments on atherosclerosis development rather than the established disease. Also, results of these studies are often confounding because of the use of superficial lesion analyses and a lack of characterization of lesion cell populations. To help overcome these translational hurdles, we propose an increased reliance on intervention models that employ investigation of changes in cellular composition at a single cell level by immunofluorescent staining and confocal microscopy. To this end, we describe a protocol for testing a putative therapeutic agent in a murine intervention model including a systematic approach for animal dissection, embedding, sectioning, staining, and quantification of brachiocephalic artery lesions. In addition, due to the phenotypic diversity of cells within late-stage atherosclerotic lesions, we describe the importance of using cell-specific, inducible lineage tracing mouse systems and how this can be leveraged for unbiased characterization of atherosclerotic lesion cell populations. Together, these strategies may assist vascular biologists to more accurately model therapeutic interventions and analyze atherosclerotic disease and will hopefully translate into a higher rate of success in clinical trials.

We propose a standardized protocol to characterize the cellular composition of late-stage murine atherosclerotic lesions including systematic methods of animal dissection, tissue embedding, sectioning, staining, and analysis of brachiocephalic arteries from atheroprone smooth muscle cell lineage tracing mice.

ניתוח כמותי של הרכב הסלולר בנגעים טרשת עורקים מתקדמת של שריר חלק תא שושלת היוחסין-עקיבה עכברים

Atherosclerosis remains the leading cause of death worldwide and, despite countless preclinical studies describing promising therapeutic targets, novel ...

Ultrasonography of the Adult Male Urinary Tract for Urinary Functional Testing

The incidence of clinical benign prostatic hyperplasia (BPH) and lower urinary tract symptoms (LUTS) is increasing due to the aging population, resulting in a significant economic and quality of life burden. Transgenic and other mouse models have been developed to recreate various aspects of this multifactorial disease; however, methods to accurately quantitate urinary dysfunction and the effectiveness of new therapeutic options are lacking. Here, we describe a method that can be used to measure bladder volume and detrusor wall thickness, urinary velocity, void volume and void duration, and urethral diameter. This would allow for the evaluation of disease progression and treatment efficacy over time. Mice were anesthetized with isoflurane, and the bladder was visualized by ultrasound. For non-contrast imaging, a 3D image was taken of the bladder to calculate volume and evaluate shape; the bladder wall thickness was measured from this image. For contrast-enhanced imaging, a catheter was placed through the dome of the bladder using a 27-gauge needle connected to a syringe by PE50 tubing. A bolus of 0.5 mL of contrast was infused into the bladder until a urination event occurred. Urethral diameter was determined at the point of the Doppler velocity sample window during the first voiding event. Velocity was measured for each subsequent event yielding a flow rate. In conclusion, high frequency ultrasound proved to be an effective method for assessing bladder and urethral measurements during urinary function in mice. This technique may be useful in the assessment of novel therapies for BPH/LUTS in an experimental setting.

We describe the use of high frequency ultrasound with contrast imaging as a method to measure bladder volume, bladder wall thickness, urine velocity, void volume, void duration, and urethral diameter. This strategy can be used to assess voiding dysfunction and treatment efficacy in various mouse models of lower urinary tract dysfunction (LUTD).

Ultrasonography של בוגרים זכר בדרכי השתן עבור בדיקות פונקציונליות השתן

The incidence of clinical benign prostatic hyperplasia (BPH) and lower urinary tract symptoms (LUTS) is increasing due to the aging population, resulting ...

Palatable Western-style Cafeteria Diet as a Reliable Method for Modeling Diet-induced Obesity in Rodents

Obesity is rapidly increasing in incidence in developed and developing countries and is known to induce or exacerbate many diseases.&#160;The health burden of obesity and its comorbid conditions highlight the need for better understanding of its pathogenesis, yet ethical constraints limit studies in humans. To this end externally valid models of obesity in laboratory animals are essential for the understanding of being overweight and obesity.&#160;While many species have been used to model the range of changes that accompany obesity in humans, rodents are most commonly used. Our laboratory has developed a western-style cafeteria diet that consistently leads to considerable weight gain and markers of metabolic disease in rodents. The diet exposes rodents to a variety of highly palatable foods to induce hyperphagia, modeling the modern western food environment. This diet rapidly induces weight gain and body fat accumulation in rats allowing for the study of effects of overeating and obesity. While the cafeteria diet may not provide the same control over macronutrient and micronutrient profile as purified high-fat or high-fat, high-sugar diets, the cafeteria diet typically induces a more severe metabolic phenotype than that observed with purified diets and is more in line with metabolic disturbances observed in the overweight and obese human population.

This protocol describes the use of a highly palatable, western-style cafeteria diet to model overeating and obesity in rodents. Here, we provide a detailed outline of food selection, preparation and measurement, and explain methodological factors that assist in generating a robust and reproducible phenotype.

טעים המערבי בסגנון הקפיטריה דיאט כשיטה אמינה עבור מידול דיאטה הנגרמת השמנה אצל מכרסמים

Obesity is rapidly increasing in incidence in developed and developing countries and is known to induce or exacerbate many diseases.&#160;The health ...

שריר חלק בשלפוחית ​​השתן רצועת התכווצות כשיטה להערכת נמוכה יותר בדרכי שתן פרמקולוגיה

שריר חלק בשלפוחית השתן רצועת התכווצות כשיטה להערכת נמוכה יותר בדרכי שתן פרמקולוגיה