Name: murine orchiectomy and ovariectomy to reduce sex hormone production
Uploaded: 2023-11-17T14:40:00
Description: Watch this Scientific Journal Video about Murine Orchiectomy and Ovariectomy to Reduce Sex Hormone Production at JoVE.com

We thank the University of Texas Southwestern Medical Center Animal Resource Center for their help in surgical training and protocol review. We thank the Wert Lab support team for their invaluable assistance. This work has been supported by funds from the National Institute of Health (NIH P30EY030413). Biorender.com was used for the creation of cartoon schematics.

All animal experiments were approved by the Institutional Animal Care and Use Committee at UTSW (APN#2019-102840).
1. Murine orchiectomy
<ol>
	<li>Prepare an aseptic working field and ensure that necessary surgical instruments have been sterilized and are readily available. Follow aseptic practices for rodent survival surgery.</li>
	<li>Record the weight of the male mouse and administer the preferred method of anesthesia according to the institutional guidelines. In this pro...

<ol>
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O., Theodorsson, A., Ingberg, E., Isaksson, I. M., Theodorsson, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ovariectomy+and+17&#946;-estradiol+replacement+in+rats+and+mice:+a+visual+demonstration.">Ovariectomy and 17&#946;-estradiol replacement in rats and mice: a visual demonstration.</a> Journal of Visualized Experiments. (64), e4013 (2012).</li><li>Valkenburg, K. C., Amend, S. R., Pienta, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+prostate+micro-dissection+and+surgical+castration.">Murine prostate micro-dissection and surgical castration.</a> Journal of Visualized Experiments. (111), e53984 (2016).</li><li>Pritchett-Corning, K. R., Luo, Y., Mulder, G. B., White, W. 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D. <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+early+bilateral+ovariectomy+in+mice+as+a+model+of+left+heart+disease.">Evaluation of early bilateral ovariectomy in mice as a model of left heart disease.</a> American Journal of Physiology Heart-Circulartory Physiology. 322 (6), H1080-H1085 (2022).</li><li>Lu, H., Ma, K., Jin, L., Zhu, H., Cao, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=17&#946;-estradiol+rescues+damages+following+traumatic+brain+injury+from+molecule+to+behavior+in+mice.">17&#946;-estradiol rescues damages following traumatic brain injury from molecule to behavior in mice.</a> Journal of Cell Physiology. 233 (2), 1712-1722 (2018).</li><li>Meydan, 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=Effects+of+testosterone+on+orchiectomy-induced+oxidative+damage+in+the+rat+hippocampus.">Effects of testosterone on orchiectomy-induced oxidative damage in the rat hippocampus.</a> Journal of Chemical Neuroanatomy. 40 (4), 281-285 (2010).</li><li>Ohlson, N., Bergh, A., Persson, M. L., Wikstr&#246;m, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Castration+rapidly+decreases+local+insulin-like+growth+factor-1+levels+and+inhibits+its+effects+in+the+ventral+prostate+in+mice.">Castration rapidly decreases local insulin-like growth factor-1 levels and inhibits its effects in the ventral prostate in mice.</a> Prostate. 66 (16), 1687-1697 (2006).</li><li>Tehranipour, M., Moghimi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuroprotective+effects+of+testosterone+on+regenerating+spinal+cord+motoneurons+in+rats.">Neuroprotective effects of testosterone on regenerating spinal cord motoneurons in rats.</a> Journal of Motor Behavior. 42 (3), 151-155 (2010).</li><li>Yamada, 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=The+impact+of+ovariectomy+on+olfactory+neuron+regeneration+in+mice.">The impact of ovariectomy on olfactory neuron regeneration in mice.</a> Chemical Senses. 45 (3), 203-209 (2020).</li><li>Koebele, S. V., Bimonte-Nelson, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+menopause:+The+utility+of+rodents+in+translational+behavioral+endocrinology+research.">Modeling menopause: The utility of rodents in translational behavioral endocrinology research.</a> Maturitas. 87, 5-17 (2016).</li></ol>

Surgical removal of testes and ovaries allows for studying murine physiology under controlled hormone deprivation. This technique is important for many fields of science, including neurodegeneration, mineral metabolism, cardiovascular, and reproductive health15,16,17,18,19,20,21. Here, we de...

The testes and ovaries are the primary organs responsible for&#160;sex hormone production. The cascade of hormonal communication leading to the production of testosterone and estrogen is a well-characterized process that begins in the hypothalamus with the release of gonadotropin-releasing hormone (GnRH)1. The release of GnRH causes the release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) from the pituitary gland. As these hormones enter the bloodstream, they then affect other tissues in the body. The primary target of LH is the testes (in males) and the ovaries (in females)2. In response to LH, the testes produce and release testosterone3. Similarly, the ovaries produce estrogen4. While the intended effects of these hormones are to prepare the cells and body for fertilization and ensure a functioning reproductive system, many other bodily systems can be affected.
Sex hormones have been linked to several physiological functions. For instance, estrogen helps maintain bone homeostasis by preventing the resorption of bone by osteoclasts. For this reason, ovariectomized mouse models can be used to study the physiology of bone diseases such as osteoporosis5,6,7. Testosterone and estrogen are also research targets for many cardiovascular and neurodegenerative diseases. Recently, elevated testosterone production coupled with high-fat diet has been linked to vascular oxidative stress8. In the brain, changes in LH after ovariectomy have caused alterations in spatial memory9. Reduction of estrogens following ovariectomy has also become a model system for studying cell death in the hippocampus, as this can induce apoptosis, resulting in memory deficits10. Testosterone has also shown a role in the growth of kidneys both in mouse models and humans following kidney transplantation11.
The creation of a hormone-deprived murine model allows for the study of sex hormones and their hormone cascades on various diseases or tissues. This can be accomplished by surgical removal of the testes (orchiectomy) or ovaries (ovariectomy). This procedure can be performed in mice of any strain when they are as young as weaning age (twenty-one days) or any adult age. Ovariectomy is performed in female mice, while orchiectomy is performed in male mice. By removing these organs, the levels of estrogen and testosterone, and many of their derivatives, such as progesterone, can be greatly reduced12,13. The process of performing orchiectomies or ovariectomies in mice can be rapid and minimally invasive with the proper technique. Rapid excision of these organs in a safe and efficient manner can allow for quick surgical processing while keeping mouse numbers minimal by having a 100% survival rate when performed correctly. Here, we detail a protocol for the rapid excision of the testes and ovaries and demonstrate the proper post-surgical monitoring to enable researchers to perform this surgery quickly and safely. We also include visual examples of the sex organs and surrounding tissues to provide the surgeon with anatomical landmarks when performing this procedure.

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			<td>Optional, can provide better clarity during procedure</td>
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			<td>Gauze Sponges, 4&#34;x4&#34;</td>
			<td>Fisher Scientific</td>
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			<td>4-0 or 5-0 Absorbable sutures are best</td>
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murine orchiectomy and ovariectomy to reduce sex hormone production

Sex hormone signaling plays a critical role in multiple organ systems as well as in the progression of various diseases, including neurodegenerative disease. The manipulation of sex hormone levels in the murine model system allows for the study of their impact on organs/tissues and within disease progression. Orchiectomy - the surgical removal of the testes - and ovariectomy - the surgical removal of the ovaries - provide a method to deplete the endogenous sex hormones so that the precise hormone levels can be provided through drug or other delivery methods. Here, we provide rapid and minimally invasive methods for both orchiectomy and ovariectomy in the murine model system for the reduction of sex hormones. This protocol details the surgical preparation and excision of the testes through the scrotal sac, and excision of the ovaries via two incisions in the right and left lateral dorsum.

The procedure presented here is performed in one- to three-month-old mice in the C57BL/6J background. Male mice weighed 16-28 g, and female mice weighed 14-24 g at the time of the procedure. This procedure has been optimized to be applicable for mice of many ages, from weaning through adulthood.
Surgical orchiectomy involves a single skin incision in the ventral scrotal sac, as depicted in Figure 1A. Both testes are removed one at a time and are severed through th...

Watch this Scientific Journal Video about Murine Orchiectomy and Ovariectomy to Reduce Sex Hormone Production at JoVE.com

Murine Orchiectomy and Ovariectomy to Reduce Sex Hormone Production

This manuscript describes a consistent way to quickly perform survival rodent orchiectomies and ovariectomies.

Sex hormone signaling plays a critical role in multiple organ systems as well as in the progression of various diseases, including neurodegenerative ...

The overall goal of this procedure is to demonstrate the surgical removal of murine testes and ovaries in a fast, safe, and effective manner. Murine orchiectomy is performed by a ventral midline incision into the scrotal sac, where the testes are isolated and removed, followed by closure of the skin incision. Murine ovariectomy is performed by a dorsolateral incision into the abdomen where the ovary is isolated and removed, followed by closure of the skin incision, with the option to repeat this procedure on the other side to perform a bilateral ovariectomy.To begin the murine orchiectomy, first start by recording the weight of a male mouse and prepare to administer the appropriate pain medication and anesthetics per your institutional guidelines. Once the mouse is under anesthesia, cover the eyes with a lubricating eye gel to protect them from drying out during the procedure. Before proceeding, ensure that the mouse is at an appropriate plane of anesthesia by performing a toe-pinch response test.Next, shave the inguinal and scrotal areas of the mouse. Using a cotton-tipped applicator, apply a thin layer of depilatory hair removal cream to the surgical site and surrounding areas. After 30 seconds, remove the depilatory cream with the clean gauze sponge.Transfer the mouse to a surgical area and make sure a source of heat is available. Use soft adhesive tape to adhere the mouse's feet to the surgical mat. If the testes have ascended into the abdomen, use gentle pressure on the abdomen to help them descend.Scrub the skin with Betadine solution, starting in the center over the surgical site and working readily outwards towards the border of the shaved area. Follow the Betadine scrub with an alcohol scrub performed in a similar manner. Repeat this process three times to ensure proper skin disinfection.Cover the mouse with the surgical drape. Grasp the skin and make a midline incision about 1 to 1 1/2 centimeters in length. Grasping the cut border of the skin, use a blunt-tipped instrument to separate the skin from the underlying tissue.Take time to do this on the lateral, superior, and inferior borders of the incision. Next, isolate one testicle by sliding the skin incision to be directly atop it. Use a pair of forceps to grasp the thin, transparent cremaster muscle that surrounds the testicle.Identification of the correct tissue can be confirmed by its transparency and its ability to move independently from the underlying tissues enclosed within it. Next, incise the cremaster muscle by making an incision into the apex of the tissue. Place a locking clamp on the posterior border of the cut cremaster muscle to hold tension on the tissue.Use a pair of forceps to grasp the testicle and exteriorize it from its cremaster sheath. Once the testicle is out, roll the tissue laterally and localize the cremaster insertion point that connects the cremaster muscle to the testicle and epididymis on the posterior side of the testicle. Clamp and tear this tissue to sever this connection.Exteriorize the inguinal fat pad by slowly pulling it out. While doing this, watch for signs of bleeding and be sure to not harm any of the blood vessels in the process. Next, use a pair of locking hemostats to clamp the spermatic cord and blood vessels.While holding slight tension on the testicle, cauterize the spermatic cord and blood vessels distal to the hemostat clamp. Slowly release the hemostat and ensure the tissue stump has good hemostasis and does not continue to bleed. Applying a slight outward pressure on the cremaster muscle will allow this tissue stump to retract into the body cavity.Bring the two sides of the cremaster muscle together. Using 4-0 or 5-0 absorbable sutures, place a suture to join the two ends of the cremaster muscle. Once the suture is finished, trim the ends of the suture material to be 1/2 a centimeter in length.Repeat this process for the other testicle by sliding the skin incision to be atop the other side, and follow the same steps used to remove the first testicle. Once both testicles have been removed and both sides of the cremaster have been sutured, evert the skin upwards and prepare to apply a wound clip to the area. Be sure the suture ends will not be caught in the wound clip.The mouse is now ready to continue with postoperative monitoring. To prepare for murine ovariectomy, weigh a female mouse and administer pain medication and anesthesia per your institutional guidelines. Administer lubricating eye gel and check to make sure the mouse has no toe-pinch response.Shave the dorsolateral regions of the mouse's back to prepare the surgical site. Using a cotton-tipped applicator, apply a layer of depilatory hair removal cream to the shaved area. After 30 seconds, remove the cream with a clean gauze sponge.Transfer the mouse to a surgical mat and ensure that a source of heat is available. Scrub the skin with Betadine solution, starting in the center and working readily outwards towards the border of the shaved area. Follow this by scrubbing the skin with 70%alcohol solution applied in a similar manner.Repeat this process for a total of three times to ensure proper skin disinfection. Plan the surgical incision as detailed in the written protocol. Place a surgical drape over the mouse's surgical site.Grasp the skin with forceps and incise the skin with a one-centimeter incision. Use a blunt-tipped instrument or a plain pair of hemostats to separate the skin from the underlying tissue. Take time to do this on the lateral, superior, and inferior borders of the incision.Grasp the abdominal wall and incise with a 1/2-centimeter incision. Use a pair of forceps to locate the ovary within the incision and pull to exteriorize it. Once the tissues have been identified, use a pair of hemostats to clamp and crush the distal uterine horn.Use a second pair of hemostats or a pair of forceps to clamp just distal to the first clamp. Holding the bottom hemostat still, pull the top clamp up and away from the body to perform a crush and tear of the distal uterine horn. Slowly release the hemostat and watch to make sure there is not excessive bleeding coming from the surgical site.Bring the two sides of the abdominal wall together and suture the incision closed, using a single suture with 4-0 absorbable suture material. Once the suture is secured, trim the ends of the suture material to be 1/2 a centimeter in length. Bring the two sides of the skin back together and prepare to place a wound clip.While everting the skin, place a wound clip over the incision site. The mouse is now ready to continue with postoperative monitoring. Document the procedure date and time and analgesic administration per your institutional guidelines.Once a mouse has undergone surgical orchiectomy or ovariectomy, transfer them to a clean, heated cage. Once a mouse has regained sternal recumbency, place them in a clean cage and provide moist chow and water. During their recovery, frequently check on the mouse's surgical site and look for any signs of infection or bleeding.To prevent damage to the wound clip, remove any rodent igloo habitats from the cage of an ovariectomy mouse for one week following surgery. After 14 days, remove the wound clips from the mouse's skin. The demonstrated protocol details how to perform an orchiectomy and ovariectomy in a mouse to generate a sex-hormone-depleted model.This involves removing the testicles through a midline incision in the scrotum, followed by exteriorizing the testicles and severing the spermatic cord in blood vessels, resulting in the removal of the testicle, epididymis, and inguinal fat pads. Surgical ovariectomy can be performed by a dorsolateral incision into the dorsum and exteriorizing the ovary, oviduct, and distal uterine horn. This is followed by severing the distal uterine horn, leading to removal of the ovary and oviduct.Careful surgical technique and proper postoperative care will result in well-healed surgical sites. These surgical procedures can be further validated by analyzing hormone levels in mouse serum samples one week after orchiectomy. Ovariectomy can be confirmed by the observation of uterine atrophy following the removal of the ovaries.This protocol enables the rapid and effective surgical removal of ovaries and testes from mice to generate a sex-hormone-depleted model. This can have many uses to study how depletion of these hormones affects various organs and physiological processes.

murine-orchiectomy-and-ovariectomy-to-reduce-sex-hormone-production

Research

JoVE Journal

Medicine

The Measurement and Treatment of Suppression in Amblyopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current estimates put the prevalence of amblyopia at approximately 1-3%1-3, the majority of cases being monocular2. Amblyopia is most frequently caused by ocular misalignment (strabismus), blur induced by unequal refractive error (anisometropia), and in some cases by form deprivation.
Although amblyopia is initially caused by abnormal visual input in infancy, once established, the visual deficit often remains when normal visual input has been restored using surgery and/or refractive correction. This is because amblyopia is the result of abnormal visual cortex development rather than a problem with the amblyopic eye itself4,5 . Amblyopia is characterized by both monocular and binocular deficits6,7 which include impaired visual acuity and poor or absent stereopsis respectively. The visual dysfunction in amblyopia is often associated with a strong suppression of the inputs from the amblyopic eye under binocular viewing conditions8. Recent work has indicated that suppression may play a central role in both the monocular and binocular deficits associated with amblyopia9,10 .
Current clinical tests for suppression tend to verify the presence or absence of suppression rather than giving a quantitative measurement of the degree of suppression. Here we describe a technique for measuring amblyopic suppression with a compact, portable device11,12 . The device consists of a laptop computer connected to a pair of virtual reality goggles. The novelty of the technique lies in the way we present visual stimuli to measure suppression. Stimuli are shown to the amblyopic eye at high contrast while the contrast of the stimuli shown to the non-amblyopic eye are varied. Patients perform a simple signal/noise task that allows for a precise measurement of the strength of excitatory binocular interactions. The contrast offset at which neither eye has a performance advantage is a measure of the "balance point" and is a direct measure of suppression. This technique has been validated psychophysically both in control13,14 and patient6,9,11 populations.
In addition to measuring suppression this technique also forms the basis of a novel form of treatment to decrease suppression over time and improve binocular and often monocular function in adult patients with amblyopia12,15,16 . This new treatment approach can be deployed either on the goggle system described above or on a specially modified iPod touch device15.

Amblyopia is a developmental disorder of the visual cortex that is often accompanied by strong suppression of one eye. We present a new technique for measuring and treating interocular suppression in patients with amblyopia that can be deployed using virtual reality goggles or a portable iPod Touch device.

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

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

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

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

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

Isolating and Using Sections of Bovine Mesenteric Artery and Vein as a Bioassay to Test for Vasoactivity in the Small Intestine

Mammalian gastrointestinal systems are constantly exposed to compounds (desirable and undesirable) that can have an effect on blood flow to and from that system. Changes in blood flow to the small intestine can result in effects on the absorptive functions of the organ. Particular interest in toxins liberated from feedstuffs through fermentative and digestive processes has developed in ruminants as an area where productive efficiencies could be improved. The video associated with this article describes an in vitro bioassay developed to screen compounds for vasoactivity in isolated cross-sections of bovine mesenteric artery and vein using a multimyograph. Once the blood vessels are mounted and equilibrated in the myograph, the bioassay itself can be used: as a screening tool to evaluate the contractile response or vasoactivity of compounds of interest; determine the presence of receptor types by pharmacologically targeting receptors with specific agonists; determine the role of a receptor with the presence of one or more antagonists; or determine potential interactions of compounds of interest with antagonists. Through all of this, data are collected real-time, tissue collected from a single animal can be exposed to a large number of different experimental treatments (an in vitro advantage), and represents vasculature on either side of the capillary bed to provide an accurate picture of what could be happening in the afferent and efferent blood supply supporting the small intestine.

The small intestine is frequently exposed to toxins that can influence blood flow and negatively impact nutrient absorption. Using a multimyograph and mesenteric artery and vein isolates, compounds or toxins of interest can be screened for vasoactivity.

Mammalian gastrointestinal systems are constantly exposed to compounds (desirable and undesirable) that can have an effect on blood flow to and from that ...

An Ex vivo Model to Study Hormone Action in the Human Breast

The study of hormone action in the human breast has been hampered by lack of adequate model systems. Upon in vitro culture, primary mammary epithelial cells tend to lose hormone receptor expression. Widely used hormone receptor positive breast cancer cell lines are of limited relevance to the in vivo situation. Here, we describe an ex vivo model to study hormone action in the human breast. Fresh human breast tissue specimens from surgical discard material such as reduction mammoplasties or mammectomies are mechanically and enzymatically digested to obtain tissue fragments containing ducts and lobules and multiple stromal cell types. These tissue microstructures kept in basal medium without growth factors preserve their intercellular contacts, the tissue architecture, and remain hormone responsive for several days. They are readily processed for RNA and protein extraction, histological analysis or stored in freezing medium. Fluorescence activated cell sorting (FACS) can be used to enrich for specific cell populations. This protocol provides a straightforward, standard approach for translational studies with highly complex, varied human specimens.

An Ex vivo Model to Study Hormone Action in the Human Breast

We have developed a novel ex vivo model to study hormone action in the human breast. It is based on tissue microstructures isolated from surgical breast tissue specimens which preserve tissue architecture, intercellular interactions, and paracrine signaling.

The study of hormone action in the human breast has been hampered by lack of adequate model systems. Upon in vitro culture, primary mammary epithelial ...

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

Single institution and population-based studies highlight that infectious complications following transrectal ultrasound guided prostate needle biopsy (TRUS PNB) are increasing. Such infections are largely attributable to quinolone resistant microorganisms which colonize the rectal vault and are translocated into the bloodstream during the biopsy procedure. A povidone iodine rectal preparation (PIRP) at time of biopsy is a simple, reproducible method to reduce rectal microorganism colony counts and therefore resultant infections following TRUS PNB.
All patients are administered three days of oral antibiotic therapy prior to biopsy. The PIRP technique involves initially positioning the patient in the standard manner for a TRUS PNB. Following digital rectal examination, 15 ml&#160;of a 10% solution of commercially available povidone iodine is mixed with 5 ml&#160;of 1% lidocaine jelly to create slurry. A 4 cm x 4 cm sterile gauze is soaked in this slurry and then inserted into the rectal vault for 2 min after which it is removed. Thereafter, a disposable cotton gynecologic swab is used to paint both the perianal area and the rectal vault to a distance of 3 cm from the anus. The povidone iodine solution is then allowed to dry for 2 - 3 min prior to proceeding with standard transrectal ultrasonography and subsequent biopsy.
This PIRP technique has been in practice at our institution since March of 2012 with an associated reduction of post-biopsy infections from 4.3% to 0.6% (p = 0.02). The principal advantage of this prophylaxis regimen is its simplicity and reproducibility with use of an easily available, inexpensive agent to reduce infections. Furthermore, the technique avoids exposing patients to additional systemic antibiotics with potential further propagation of multi-drug resistant organisms. Usage of PIRP at TRUS PNB, however, is not applicable for patients with iodine or shellfish allergies.

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.

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

Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of clinical efficacy, they are until now imperfect solutions. Stem cell therapy using ASC sheets could provide a solution to this problem. ASCs are considered as promising candidates for promoting tissue healing because of their trophic and immunomodulatory properties. Here, we provide methods to produce high-density ASC sheets, that are transplanted onto a colorectal anastomosis in a rat model to reduce the leakage. ASCs formed cell sheets in thermo-responsive culture dishes that could be easily detached. On the day of the transplantation, a partial colectomy with a 5-suture colorectal anastomosis was performed. Animals were immediately transplanted with 1 ASC sheet per rat. ASC sheets adhered spontaneously to the anastomosis without any glue, suture, or any biomaterial. Animal groups were sacrificed 3 and 7 days postoperatively. Compared to transplanted animals, the incidence of anastomotic abscesses and leakage was higher in control animals. In our model, the transplantation of ASC sheets after colorectal anastomosis was successful and associated with a lower leakage rate.

The preparation and transplantation of adipose tissue derived stem cell (ASC) sheets onto insufficiently sutured colorectal anastomoses in a rat model is presented. This novel application shows that ASC sheets can reduce colorectal anastomosis leakage.

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

A Clinical Trial Assessing the Safety, Efficacy, and Delivery of Olive-Oil-Based Three-Chamber Bags for Parenteral Nutrition

Limited evidence exists to precisely estimate efficacy and safety differences between parenteral nutrition (PN) prepared using olive-oil-based three-chamber bags (3CBs) and soybean-oil-based compounded bags (CoBs) in hospitalized adult patients. We designed a multicenter, randomized, prospective, open-label, noninferiority protocol to compare the efficacy, safety, and distribution of olive-oil-based 3CBs and soybean-oil-based CoB formulations in adult Chinese patients requiring PN during surgical intervention. Subjects were randomized to receive either one of the study treatments using an interactive voice or web-based recognition system in accordance with the randomization code. Randomization was further stratified based on the study site and surgical category. Both treatment groups received similar amounts of calories and protein. In addition, the two study treatments contained a similar composition of the amino-acid component. The only difference between the two PN formulations was the lipid constitution. The duration of administration of study treatments was a minimum of 5 days up to a maximum of 14 days after the surgical procedure. The primary efficacy endpoint was serum prealbumin levels on day 5 of the study. Noninferiority was proved if the anti-log of the lower bound of the 95% confidence interval (CI) of the treatment difference was at least 0.80. Other efficacy measures included treatment preparation time; duration to achieve tolerability of oral nutrition; associated infectious complications; length of hospitalization; and laboratory assessment of markers of nutrition, inflammation, metabolism, and oxidative stress. A total of 458 patients were enrolled in the study. The results showed that olive-oil-based 3CBs were non-inferior to soybean-based CoBs, besides being well tolerated. The infection rate was found to be significantly lower in the olive-oil-based 3CB group. Thus, this study may be used as a reference for future research on lipid emulsion and 3CBs.

Here, we present a protocol for a study comparing the efficacy, safety, and delivery of olive-oil-based 3CB and soybean-oil-based CoB formulations in adults requiring parenteral nutrition. The results revealed that olive-oil-based 3CBs is non-inferior and well tolerated compared to soybean formulations.

Limited evidence exists to precisely estimate efficacy and safety differences between parenteral nutrition (PN) prepared using olive-oil-based ...

Mechanisms Underlying Gut Hormone Secretion Using the Isolated Perfused Rat Small Intestine

The gut is the largest endocrine organ of the body, producing more than 15 different peptide hormones that regulate appetite and food intake, digestion, nutrient absorption and distribution, and post-prandial glucose excursions. Understanding the molecular mechanisms that regulate gut hormone secretion is fundamental for understanding and translating gut hormone physiology. Traditionally, the mechanisms underlying gut hormone secretion are either studied in vivo (in experimental animals or humans) or using gut hormone-secreting primary mucosal cell cultures or cell lines. Here, we introduce an isolated perfused rat small intestine as an alternative method for studying gut hormone secretion. The virtues of this model are that it relies on the intact gut, meaning that it recapitulates most of the physiologically important parameters responsible for the secretion in in vivo studies, including mucosal polarization, paracrine relationships and routes of perfusion/stimulus exposure. In addition, and unlike in vivo studies, the isolated perfused rat small intestine allows for almost complete experimental control and direct assessment of secretion. In contrast to in vitro studies, it is possible to study both the magnitude and the dynamics of secretion and to address important questions, such as what stimuli cause secretion of different gut hormones, from which side of the gut (luminal or vascular) is secretion stimulated, and to analyze in detail molecular sensors underlying the secretory response. In addition, the preparation is a powerful model for the study of intestinal absorption and details regarding the dynamics of intestinal absorption including the responsible transporters.

Here, we present a powerful and physiological model to study the molecular mechanisms underlying gut hormone secretion and intestinal absorption &#8212; the isolated perfused rat small intestine.

The gut is the largest endocrine organ of the body, producing more than 15 different peptide hormones that regulate appetite and food intake, digestion, ...

Cooling or Warming the Esophagus to Reduce Esophageal Injury During Left Atrial Ablation in the Treatment of Atrial Fibrillation

Ablation of the left atrium using either radiofrequency (RF) or cryothermal energy is an effective treatment for atrial fibrillation (AF) and is the most frequent type of cardiac ablation procedure performed. Although generally safe, collateral injury to surrounding structures, particularly the esophagus, remains a concern. Cooling or warming the esophagus to counteract the heat from RF ablation, or the cold from cryoablation, is a method that is used to reduce thermal esophageal injury, and there are increasing data to support this approach. This protocol describes the use of a commercially available esophageal temperature management device to cool or warm the esophagus to reduce esophageal injury during left atrial ablation. The temperature management device is powered by standard water-blanket heat exchangers, and is shaped like a standard orogastric tube placed for gastric suctioning and decompression. Water circulates through the device in a closed-loop circuit, transferring heat across the silicone walls of the device, through the esophageal wall. Placement of the device is analogous to the placement of a typical orogastric tube, and temperature is adjusted via the external heat-exchanger console.

The goal of this protocol is to describe the use of esophageal temperature modulation to counteract esophageal thermal injury from left atrial ablation for the treatment of atrial fibrillation.

Ablation of the left atrium using either radiofrequency (RF) or cryothermal energy is an effective treatment for atrial fibrillation (AF) and is the most ...

Network Pharmacology Prediction and Experimental Validation of Trichosanthes-Fritillaria thunbergii Action Mechanism Against Lung Adenocarcinoma

We aimed to study the mechanism of Trichosanthes-Fritillaria thunbergii in treating lung adenocarcinoma (LUAD) based on network pharmacology and experimental verification. The effective components and potential targets of Trichosanthis and Fritillaria thunbergii were collected by high-throughput experiment and reference-guided (HERB) database of traditional Chinese medicine and a similarity ensemble approach (SEA) database, and the LUAD-related targets were queried by the GeneCards and Online Mendelian Inheritance in Man (OMIM) databases. A drug-component-disease-target network was constructed by Cytoscape software. Protein-protein interaction (PPI) network, gene ontology (GO) function, and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses were conducted to obtain core targets and key pathways. An aqueous extract of Trichosanthes-Fritillaria thunbergii and A549 cells were used for the subsequent experimental validation. Through the HERB database and literature search, 31 effective compounds and 157 potential target genes of Trichosanthes-Fritillaria thunbergii were screened, of which 144 were regulatory targets of Trichosanthes-Fritillaria thunbergii in the treatment of lung adenocarcinoma. The GO functional enrichment analysis showed that the mechanism of action of Trichosanthes-Fritillaria thunbergii against lung adenocarcinoma is mainly protein phosphorylation. The KEGG pathway enrichment analysis suggested that the treatment of lung adenocarcinoma by Trichosanthes-Fritillaria thunbergii mainly involves the PI3K/AKT signaling pathway. The experimental validation showed that an aqueous extract of Trichosanthes-Fritillaria thunbergii could inhibit the proliferation of A549 cells and the phosphorylation of AKT. Through network pharmacology and experimental validation, it was verified that the PI3K/AKT signaling pathway plays a vital role in the action of Trichosanthes-Fritillaria thunbergii in treating lung adenocarcinoma.

Network Pharmacology Prediction and Experimental Validation of Trichosanthes-Fritillaria thunbergii Action Mechanism Against Lung Adenocarcinoma

This study reveals the mechanism of Trichosanthes-Fritillaria thunbergii in treating lung adenocarcinoma based on network pharmacology and experimental verification. The study also demonstrates that the PI3K/AKT signaling pathway plays a vital role in the action of Trichosanthes-Fritillaria thunbergii in treating lung adenocarcinoma.