The authors have no acknowledgements.

1. Installation of Dynamic Tissue System
<ol>
	<li>Exclude if there is hemodynamic instability, a need for further abdominal washouts, or a concern for intraabdominal sepsis.
	<ol>
		<li>If an ostomy exists, trim the ostomy wafer to make as narrow as possible and outline on abdominal skin. Cover ostomy with 4x4 and secure in place with adhesive transparent film dressing.</li>
		<li>Apply antimicrobial betadine impregnated adhesive drape to the skin of the abdomen widely, covering the operating field.</li>
		<li>Wash the abdomen out with at least 2 L of 40.5&#176; C normal saline.</li>
		<li>Measure and record the myofascial gap (MFG), visceral extrusion (VE), and the length of incision in centimeters for documentation of progress on the operative note.</li>
		<li>Mark the skin of the anterior abdominal wall 5 cm laterally from medial fascial edge to create an ellipse around the midline wound.</li>
		<li>Place the elastomer buttons side by side on the ellipse created in step 1.1.5. with the &#8220;U&#8221; of the elastomer at the 5 cm ellipse demarcation around the midline wound. This will assure 3 cm spacing as recommended.</li>
		<li>Create superficial thickness dermotomies with an 11 blade.</li>
		<li>Pull the elastomer through the abdominal wall using the cannulator.</li>
		<li>Repeat the same steps on the opposite side. This time pull the elastomer across the myofascial gap and secure both ends with a hemostat.</li>
		<li>Loosely attach the elastomer bands to the elastomer buttons. Do not tighten elastomer bands.</li>
		<li>Attach elastomer adhesion stickers on the button tails.</li>
		<li>Perform osteopathic maneuvers throughout installation. This is performed with two people standing on either side of the patient. Apply bimanual forces in the flank area, gently massaging towards the midline and create circular motions synchronized with the other operator first towards the head and then creating circular motions towards the feet.</li>
	</ol>
	</li>
</ol>
2. Silicone Visceral Protector Inserted
<ol>
	<li>Insert silicone visceral protector midline over abdominal viscera, taking care to protect any ostomies (Figure 3).</li>
	<li>Envelop the abdominal viscera with silicone visceral protector down to each paracolic gutter.</li>
	<li>Create necessary cut outs in the visceral protector with scissors to protect the ostomies.</li>
	<li>Place elastomer retainer in the midline on top of visceral protector and evenly space elastomer bands into the retainer (trim to size).</li>
	<li>Adjust elastomer bands slowly in between osteopathic maneuvers.</li>
	<li>Stop elastomer adjustments when tension hashmarks across the MFG is 1.5 &#8211; 2x stretch.</li>
	<li>Record the MFG and VE with a ruler in centimeters at the start, install, and completion (Figure 4).</li>
</ol>
3. Installation of Negative Pressure Wound Therapy Device
<ol>
	<li>Make NPWT device sponge as thing as possible and place in the midline above the elastomer retainer (Figure 5).</li>
	<li>Cover with vacuum drape tape and apply the negative pressure device with continuous suction at -100 mmHg.</li>
	<li>Perform osteopathic maneuver a few more times.</li>
</ol>
4. Elastomer Adjustment
<ol>
	<li>Bring the patient back to operating room, or if your facility has the capability to perform in ICU, periodically. First elastomer adjustment can be 2 &#8211; 5 days after installation.</li>
	<li>Remove the NPWT device overlying the midline wound with scissors, taking care to leave the adhesive iodine drape on the skin intact.</li>
	<li>Prep the abdomen widely including the elastomers in place with 4% chlorhexidine gluconate.</li>
	<li>Perform osteopathic maneuvers intermittently throughout procedure. See step 1.1.12. for details.</li>
	<li>Record the MFG and myofascial apposition (Figure 6).</li>
	<li>Irrigate the midline wound with at least 2 L of 40.5&#176; C normal saline, taking care not to spill water on the DTS or the antimicrobial iodine impregnated adhesive drape.</li>
	<li>Adjust elastomers by releasing the elastomer from the anchor and pulling each elastomer laterally, away from the midline.</li>
	<li>Reapply the black sponge NPWT device.</li>
</ol>
5. Fascial Closure
<ol>
	<li>Bring the patient back to the operating room. Prep widely.</li>
	<li>Remove the elastomer retainer.</li>
	<li>Remove the visceral protector and irrigate the abdominal cavity with at least 2 L of normal saline.</li>
	<li>Close the fascia in a bi-directional fashion utilizing a Smead-Jones technique and a #2 vicryl suture on a TP1 needle.</li>
	<li>Irrigate and dry the midline wound.</li>
	<li>Apply PUBM micromatrix powder to the midline wound and evenly distribute the powder so that the newly created linea alba is covered in PUBM micromatrix.</li>
	<li>Hydrate a two-layer, 10x15 cm, PUBM wound sheet and delaminate so there are 2 single sheets. Apply the sheets on top of the powder to cover the entire midline wound surface (Figure 7).</li>
	<li>Appose skin edge with strips of vac drape tape (Figure 8).</li>
</ol>

<ol>
	<li>Kirkpatrick, A. W., 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=Update+from+the+Abdominal+Compartment+Society+(WSACS)+on+intra-abdominal+hypertension+and+abdominal+compartment+syndrome:+past,+present,+and+future+beyond+Banff+2017.">Update from the Abdominal Compartment Society (WSACS) on intra-abdominal hypertension and abdominal compartment syndrome: past, present, and future beyond Banff 2017.</a> Anaesthesiology Intensive Therapy. 49 (2), 83-87 (2017).</li><li>Yetisir, F., Sarer, A. E., Acar, H. Z., Aygar, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delayed+Closure+of+61+Open+Abdomen+Patients+Based+on+an+Algorithm.">Delayed Closure of 61 Open Abdomen Patients Based on an Algorithm.</a> Indian Journal of Surgery. 79 (1), 38-44 (2017).</li><li>Ribeiro Junior, M. A., 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=Open+abdomen+in+gastrointestinal+surgery:+Which+technique+is+the+best+for+temporary+closure+during+damage+control?.">Open abdomen in gastrointestinal surgery: Which technique is the best for temporary closure during damage control?.</a> World Journal of Gastrointestinal Surgery. 8 (8), 590-597 (2016).</li><li>Karakose, O., 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=Bogota+Bag+Use+in+Planned+Re-Laparotomies.">Bogota Bag Use in Planned Re-Laparotomies.</a> Medical Science Monitor. 22, 2900-2904 (2016).</li><li>Lassalle, S., Chechin, C., de la Forge, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+cleansing+technique+for+complex+wounds.">A new cleansing technique for complex wounds.</a> Soins. 62 (814), 12-15 (2017).</li><li>Strang, S. G., Van Lieshout, E. M., Van Waes, O. J., Verhofstad, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevalence+and+mortality+of+abdominal+compartment+syndrome+in+severely+injured+patients:+A+systematic+review.">Prevalence and mortality of abdominal compartment syndrome in severely injured patients: A systematic review.</a> The Journal of Trauma and Acute Care Surgery. 81 (3), 585-592 (2016).</li><li>Bressan, A. K., Ball, C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intra-abdominal+hypertension+and+abdominal+compartment+syndrome+in+acute+pancreatitis,+hepato-pancreato-biliary+operations+and+liver+transplantation.">Intra-abdominal hypertension and abdominal compartment syndrome in acute pancreatitis, hepato-pancreato-biliary operations and liver transplantation.</a> Anaesthesiology Intensive Therapy. 49 (2), 159-166 (2017).</li><li>Muresan, M., 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=How+much+does+decompressive+laparotomy+reduce+the+mortality+rate+in+primary+abdominal+compartment+syndrome?:+A+single-center+prospective+study+on+66+patients.">How much does decompressive laparotomy reduce the mortality rate in primary abdominal compartment syndrome?: A single-center prospective study on 66 patients.</a> Medicine (Baltimore). 96 (5), e6006 (2017).</li><li>Coccolini, F., 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=IROA:+International+Register+of+Open+Abdomen,+preliminary+results.">IROA: International Register of Open Abdomen, preliminary results.</a> World Journal of Emergency Surgery. 12, 8 (2017).</li><li>Lecheminant, J., Field, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Porcine+urinary+bladder+matrix:+a+retrospective+study+and+establishment+of+protocol.">Porcine urinary bladder matrix: a retrospective study and establishment of protocol.</a> Journal of Wound Care. 21 (10), 482 (2012).</li><li>Coccolini, F., 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=IROA:+International+Register+of+Open+Abdomen,+preliminary+results.">IROA: International Register of Open Abdomen, preliminary results.</a> World Journal of Emergency Surgery. 12, 8 (2017).</li><li>Muturi, A., Ndaguatha, P., Ojuka, D., Kibet, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevalence+and+predictors+of+intra-abdominal+hypertension+and+compartment+syndrome+in+surgical+patients+in+critical+care+units+at+Kenyatta+National+Hospital.">Prevalence and predictors of intra-abdominal hypertension and compartment syndrome in surgical patients in critical care units at Kenyatta National Hospital.</a> BMC Emergency Medicine. 17 (1), (2017).</li><li>Okullo, A., 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+Abdominal+Reapproximation+Anchor+Device.">The Abdominal Reapproximation Anchor Device.</a> Surgical Innovation. 24 (1), 49-54 (2017).</li><li>Yetisir, F., Sarer, A. E., Acar, H. Z., Aygar, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delayed+Closure+of+61+Open+Abdomen+Patients+Based+on+an+Algorithm.">Delayed Closure of 61 Open Abdomen Patients Based on an Algorithm.</a> Indian Journal of Surgery. 79 (1), 38-44 (2017).</li><li>Chaudhry, H., Bukiet, B., Zhiming, J., Stecco, A., Findley, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deformations+experienced+in+the+human+skin,+adipose+tissue,+and+fascia+in+osteopathic+manipulative+medicine.">Deformations experienced in the human skin, adipose tissue, and fascia in osteopathic manipulative medicine.</a> The Journal of the American Osteopathic Association. 114 (1), 780-787 (2017).</li></ol>

Dr. Catherine Ronaghan is an ACell&#160;cadaver lab proctor and speaker. The rest of the authors have nothing to disclose.

The most critical step of the protocol for closing a complex abdominal wound is performing osteopathic maneuvers before elastomer placement, after elastomer placement, and before and after elastomer adjustments. In addition, we perform osteopathic maneuvers on these patients after surgery three times a day, for at least five days. Our approach describes the use of osteopathic maneuver prior and after elastomer adjustments. The anecdotal observation has been that these maneuvers aid in fascial approximation, by facilitati...

The increasing prevalence of abdominal compartment syndrome (ACS) has led to an emergence of various temporary abdominal closure (TAC) techniques1. TAC is performed to prevent evisceration, assist in the removal of unwanted intraperitoneal fluid, minimize intra-abdominal complications, and expedite the closure of the abdominal cavity2. Closure of an open abdomen facilitates restoration of normal physiology in the patient3. Prolonged duration of an open abdomen results in complications&#160;such as fistula formation and an inability to close the abdomen4. There are several methods to achieve final closure of an open abdomen.
The simplest way to temporarily close an abdomen is by using towel clips to close the skin5. One of the most commonly used and studied abdominal closure techniques is negative pressure wound therapy (NPWT)5. For the NPWT, a nonadherent barrier to protect the intraabdominal contents is applied followed by a moisture-absorbing sponge-like material,&#160;an outermost adhesive layer to sure the dressing in place, and a negative pressure mechanism6. A Bogota bag can also be used for temporary closure of an open abdomen. A Bogota bag is an empty intravenous fluid bag cut in half and sutured to&#160;skin edges7. NPWT and the Bogota bag closure are two temporizing measures that facilitate delayed primary closure of the abdominal cavity7.
Once the abdomen is deemed ready for closure, different closure methods can be utilized. The simplest way is to apply a split-thickness graft over the omentum once it has formed healthy granulation tissue. If the wound is not contaminated, a nonabsorbable synthetic sheet may be used to bridge the fascial edges8. If the fascial gap is less than 14-20 cm in maximal diameter, component separation of the rectus sheath can be performed9.
Some abdominal closure techniques allow for gradual reapproximation of the fascial edges and eventual primary closure10. A Wittmann patch consists of two opposing Velcro sheets that are sutured to each fascial edge11. The opposing sheets are then fastened together in the midline. This mechanism allows easy re-entry into the abdomen and adjustment for abdominal compartment pressures. Additionally, this can&#160;provide midline traction on the fascial edges that can prevent retraction of the fascial edges and also facilitate primary closure of the fascia.
Alternatively, a DTS is available and is part of the technique described in this paper. The described DTS is composed to a silicone viscera protector that is applied over abdominal contents to prevent adhesions and adherence of viscera to the abdominal wall. Adjustable elastomers then penetrate the full abdominal wall thickness on each side and provide medializing dynamic tension, allowing relaxation of the flat muscles (obliques and transversus abdominus). This allows medialization of the rectus myofascial units (Figure 1). A product composed of porcine urinary bladder extracellular matrix can be placed in the subcutaneous space once primary myofascial closure is achieved (Figure 2). Porcine xenograft placement in the subcutaneous space augments and expedites wound healing through angiogenesis, innervation, modulation of the inflammatory response, and resistance to infection12.
In this study, we describe a novel technique of primary abdominal closure following abdominal compartment syndrome utilizing a dynamic closure system and a biologic xenograft. At our level 1&#160;trauma and acute care center, abdominal compartment syndrome is a common diagnosis. Prior to utilization of this novel method, most catastrophic open abdomens were not amenable to primary closure and a skin graft was placed over the viscera or bridging&#160;mesh. Since the adoption of this method in May of 2016, we have closed 100% of open abdomens due to abdominal compartment syndrome in a high-risk&#160;population (average BMI 40.45, SD 9.83) (Table 1).

<table border="0" cellpadding="0" cellspacing="0" style="width:628px;" width="628">
	<tbody>
		<tr height="42">
			<td height="42" style="height:43px;width:238px;">ABRA Abdominal Wall Closure Set</td>
			<td style="width:209px;">Southmedic</td>
			<td style="width:182px;">CWK08 Abdominal</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:43px;width:238px;">3M Ioban 2 Antimicrobial Incise Drape</td>
			<td style="width:209px;">3M</td>
			<td style="width:182px;">6651EZ</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:43px;width:238px;">MicroMatrix Micronized Particles 200 mg</td>
			<td style="width:209px;">ACell</td>
			<td style="width:182px;">MM0200</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:43px;width:238px;">Cytal Wound Matrix 2-Layer 10 x 15 cm</td>
			<td style="width:209px;">ACell</td>
			<td style="width:182px;">WSM1015</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:43px;width:238px;">Negative Pressure Therapy System</td>
			<td style="width:209px;">KCI</td>
			<td style="width:182px;">09-03-193.ABT.IE</td>
		</tr>
	</tbody>
</table>

a case series of successful abdominal closure utilizing a novel technique combining a mechanical closure system with a biologic xenograft that accelerates wound healing

In the acute setting, once intra-abdominal injuries have been addressed, the next great hurdle is restoring a functional and intact abdominal compartment. The short and long-term consequences of living with a chronically open abdominal compartment include pulmonary, musculoskeletal, gastrointestinal, and emotional disability. The closure of catastrophic open abdomens presents a challenge to the surgeon. We present a technique utilizing a mechanical abdominal closure device in conjunction with biologic xenograft in closing complex open abdomens. This technique offers another option for definitive fascial closure and accelerated wound healing in this difficult patient population. The dynamic tissue system (DTS) is installed after control of original intraabdominal pathology. A porcine urinary bladder matrix (PUBM) is then placed in the subcutaneous space once fascial closure is achieved. Overall, primary myofascial closure was achieved in 100% of patients at a mean of 9.36 days.

We have analyzed a total of 11 patients so far with catastrophic open abdomens. Primary myofascial closure was achieved at a mean of 9.36 days. We had 0% surgical site infections (SSI) and achieved 100% primary myofascial closure. No enteroatmospheric fistula resulted in this technique, unless present prior to this DTS and xenograft approach. Since May 2016, zero open abdomens were left open or covered with a skin graft (Figure 9).
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Watch this Scientific Journal Video about A Case Series of Successful Abdominal Closure Utilizing a Novel Technique Combining a Mechanical Closure System with a Biologic Xenograft that Accelerates Wou

A Case Series of Successful Abdominal Closure Utilizing a Novel Technique Combining a Mechanical Closure System with a Biologic Xenograft that Accelerates Wound Healing

Closure of catastrophic open abdominal wounds presents a challenge to the surgeon. We present a surgical technique utilizing a combination of mechanical and biologic xenograft closure systems in closing complex open abdominal wounds. This technique offers another option to the surgeon for definitive fascial closure and accelerated wound healing.

In the acute setting, once intra-abdominal injuries have been addressed, the next great hurdle is restoring a functional and intact abdominal compartment. ...

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Medicine

51.1K Views.  Texas Tech University Health Sciences Center. Closure of catastrophic open abdominal wounds presents a challenge to the surgeon. We present a surgical technique utilizing a combination of mechanical and biologic xenograft closure systems in closing complex open abdominal wounds. This technique offers another option to the surgeon for definitive fascial closure and accelerated wound healing.

Video: A Case Series of Successful Abdominal Closure Utilizing a Novel Technique Combining a Mechanical Closure System with a Biologic Xenograft that Accelerates Wound Healing

A Novel Capsulorhexis Technique Using Shearing Forces with Cystotome

Purpose:
To demonstrate a capsulorhexis technique using predominantly shearing forces with a cystotome on a virtual reality simulator and on a human eye.
Method:
Our technique involves creating the initial anterior capsular tear with a cystotome to raise a flap. The flap left unfolded on the lens surface. The cystotome tip is tilted horizontally and is engaged on the flap near the leading edge of the tear. The cystotome is moved in a circular fashion to direct the vector forces. The loose flap is constantly swept towards the centre so that it does not obscure the view on the tearing edge.
Results:
Our technique has the advantage of reducing corneal wound distortion and subsequent anterior chamber collapse. The capsulorhexis flap is moved away from the tear leading edge allowing better visualisation of the direction of tear. This technique offers superior control of the capsulorhexis by allowing the surgeon to change the direction of the tear to achieve the desired capsulorhexis size.
Conclusions:
The EYESI Surgical Simulator is a realistic training platform for surgeons to practice complex capsulorhexis techniques. The shearing forces technique is a suitable alternative and in some cases a far better technique in achieving the desired capsulorhexis.

This video demonstrates a novel capsulorhexis technique in modern cataract surgery. Capsulorhexis is the most difficult and important part of phacoemulsification surgery.

Purpose:
To demonstrate a capsulorhexis technique using predominantly shearing forces with a cystotome on a virtual reality simulator and on a human eye....

Corneal Confocal Microscopy: A Novel Non-invasive Technique to Quantify Small Fibre Pathology in Peripheral Neuropathies

The accurate quantification of peripheral neuropathy is important to define at risk patients, anticipate deterioration, and assess new therapies. Conventional methods assess neurological deficits and electrophysiology and quantitative sensory testing quantifies functional alterations to detect neuropathy. However, the earliest damage appears to be to the small fibres and yet these tests primarily assess large fibre dysfunction and have a limited ability to demonstrate regeneration and repair. The only techniques which allow a direct examination of unmyelinated nerve fibre damage and repair are sural nerve biopsy with electron microscopy and skin-punch biopsy. However, both are invasive procedures and require lengthy laboratory procedures and considerable expertise. Corneal Confocal microscopy is a non-invasive clinical technique which provides in-vivo imaging of corneal nerve fibres. We have demonstrated early nerve damage, which precedes loss of intraepidermal nerve fibres in skin biopsies together with stratification of neuropathic severity and repair following pancreas transplantation in diabetic patients. We have also demonstrated nerve damage in idiopathic small fibre neuropathy and Fabry's disease.

Corneal Confocal microscopy is a non-invasive clinical technique which may be used to quantify C fibre damage to diagnose and stratify patients with increasing neuropathic severity.

The accurate quantification of peripheral neuropathy is important to define at risk patients, anticipate deterioration, and assess new therapies. ...

In vivo Dual Substrate Bioluminescent Imaging

Our understanding of how and when breast cancer cells transit from established primary tumors to metastatic sites has increased at an exceptional rate since the advent of in vivo bioluminescent imaging technologies 1-3. Indeed, the ability to locate and quantify tumor growth longitudinally in a single cohort of animals to completion of the study as opposed to sacrificing individual groups of animals at specific assay times has revolutionized how researchers investigate breast cancer metastasis. Unfortunately, current methodologies preclude the real-time assessment of critical changes that transpire in cell signaling systems as breast cancer cells (i) evolve within primary tumors, (ii) disseminate throughout the body, and (iii) reinitiate proliferative programs at sites of a metastatic lesion. However, recent advancements in bioluminescent imaging now make it possible to simultaneously quantify specific spatiotemporal changes in gene expression as a function of tumor development and metastatic progression via the use of dual substrate luminescence reactions. To do so, researchers take advantage for two light-producing luciferase enzymes isolated from the firefly (Photinus pyralis) and sea pansy (Renilla reniformis), both of which react to mutually exclusive substrates that previously facilitated their wide-spread use in in vitro cell-based reporter gene assays 4. Here we demonstrate the in vivo utility of these two enzymes such that one luminescence reaction specifically marks the size and location of a developing tumor, while the second luminescent reaction serves as a means to visualize the activation status of specific signaling systems during distinct stages of tumor and metastasis development. Thus, the objectives of this study are two-fold. First, we will describe the steps necessary to construct dual bioluminescent reporter cell lines, as well as those needed to facilitate their use in visualizing the spatiotemporal regulation of gene expression during specific steps of the metastatic cascade. Using the 4T1 model of breast cancer metastasis, we show that the in vivo activity of a synthetic Smad Binding Element (SBE) promoter was decreased dramatically in pulmonary metastasis as compared to that measured in the primary tumor 4-6. Recently, breast cancer metastasis was shown to be regulated by changes within the primary tumor microenvironment and reactive stroma, including those occurring in fibroblasts and infiltrating immune cells 7-9. Thus, our second objective will be to demonstrate the utility of dual bioluminescent techniques in monitoring the growth and localization of two unique cell populations harbored within a single animal during breast cancer growth and metastasis.

In vivo Dual Substrate Bioluminescent Imaging

Herein we describe the methods to construct, visualize, and quantify the bioluminescent reactions of both firefly and renilla luciferase enzymes expressed in metastatic breast cancer cells during their growth and metastasis in vivo.

Our understanding of how and when breast cancer cells transit from established primary tumors to metastatic sites has increased at an exceptional rate ...

Utilizing Transcranial Magnetic Stimulation to Study the Human Neuromuscular System

Transcranial magnetic stimulation (TMS) has been in use for more than 20 years 1, and has grown exponentially in popularity over the past decade. While the use of TMS has expanded to the study of many systems and processes during this time, the original application and perhaps one of the most common uses of TMS involves studying the physiology, plasticity and function of the human neuromuscular system. Single pulse TMS applied to the motor cortex excites pyramidal neurons transsynaptically 2 (Figure 1) and results in a measurable electromyographic response that can be used to study and evaluate the integrity and excitability of the corticospinal tract in humans 3. Additionally, recent advances in magnetic stimulation now allows for partitioning of cortical versus spinal excitability 4,5. For example, paired-pulse TMS can be used to assess intracortical facilitatory and inhibitory properties by combining a conditioning stimulus and a test stimulus at different interstimulus intervals 3,4,6-8. In this video article we will demonstrate the methodological and technical aspects of these techniques. Specifically, we will demonstrate single-pulse and paired-pulse TMS techniques as applied to the flexor carpi radialis (FCR) muscle as well as the erector spinae (ES) musculature. Our laboratory studies the FCR muscle as it is of interest to our research on the effects of wrist-hand cast immobilization on reduced muscle performance6,9, and we study the ES muscles due to these muscles clinical relevance as it relates to low back pain8. With this stated, we should note that TMS has been used to study many muscles of the hand, arm and legs, and should iterate that our demonstrations in the FCR and ES muscle groups are only selected examples of TMS being used to study the human neuromuscular system.

Transcranial magnetic stimulation (TMS) is a non-invasive tool to gain insight on the physiology and function of the human nervous system. Here, we present our TMS techniques to study cortical excitability of the upper limb and lumbar musculature.

Transcranial magnetic stimulation (TMS) has been in use for more than 20 years 1, and has grown exponentially in popularity over the past decade.  While ...

Intraoperative Detection of Subtle Endometriosis: A Novel Paradigm for Detection and Treatment of Pelvic Pain Associated with the Loss of Peritoneal Integrity

Endometriosis is a common disease affecting 40 to 70% of reproductive-aged women with chronic pelvic pain (CPP) and/or infertility. The purpose of this study was to demonstrate the use of a blue dye (methylene blue) to stain peritoneal surfaces during laparoscopy (L/S) to detect the loss of peritoneal integrity in patients with pelvic pain and suspected endometriosis. Forty women with CPP and 5 women without pain were evaluated in this pilot study. During L/S, concentrated dye was sprayed onto peritoneal surfaces, then aspirated and rinsed with Lactated Ringers solution. Areas of localized dye uptake were evaluated for the presence of visible endometriotic lesions. Areas of intense peritoneal staining were resected and some fixed in 2.5% buffered gluteraldehyde and examined by scanning (SEM) electron microscopy. Blue dye uptake was more common in women with endometriosis and chronic pelvic pain than controls (85% vs. 40%). Resection of the blue stained areas revealed endometriosis by SEM and loss of peritoneal cell-cell contact compared to normal, non-staining peritoneum. Affected peritoneum was associated with visible endometriotic implants in most but not all patients. Subjective pain relief was reported in 80% of subjects. Based on scanning electron microscopy, we conclude that endometrial cells extend well beyond visible implants of endometriosis and appear to disrupt the underlying mesothelium. Subtle lesions of endometriosis could therefore cause pelvic pain by disruption of peritoneal integrity, allowing menstrual or ovulatory blood and associated pain factors access to underlying sensory nerves. Complete resection of affected peritoneum may provide a better long-term treatment for endometriosis and CPP. This simple technique appears to improve detection of subtle or near invisible endometriosis in women with CPP and minimal visual findings at L/S and may serve to elevate diagnostic accuracy for endometriosis at laparoscopy.

Loss of peritoneal integrity provides a new paradigm to understand and treat chronic pelvic pain in women with mild forms of endometriosis and can be easily detected using intraoperative instillation of dye at the time of laparoscopy.

Endometriosis is a  common disease affecting 40 to 70% of reproductive-aged women with chronic  pelvic pain (CPP) and/or infertility. The purpose of this ...

Utilizing Repetitive Transcranial Magnetic Stimulation to Improve Language Function in Stroke Patients with Chronic Non-fluent Aphasia

Transcranial magnetic stimulation (TMS) has been shown to significantly improve language function in patients with non-fluent aphasia1. In this experiment, we demonstrate the administration of low-frequency repetitive TMS (rTMS) to an optimal stimulation site in the right hemisphere in patients with chronic non-fluent aphasia. A battery of standardized language measures is administered in order to assess baseline performance. Patients are subsequently randomized to either receive real rTMS or initial sham stimulation. Patients in the real stimulation undergo a site-finding phase, comprised of a series of six rTMS sessions administered over five days; stimulation is delivered to a different site in the right frontal lobe during each of these sessions. Each site-finding session consists of 600 pulses of 1 Hz rTMS, preceded and followed by a picture-naming task. By comparing the degree of transient change in naming ability elicited by stimulation of candidate sites, we are able to locate the area of optimal response for each individual patient. We then administer rTMS to this site during the treatment phase. During treatment, patients undergo a total of ten days of stimulation over the span of two weeks; each session is comprised of 20 min of 1 Hz rTMS delivered at 90% resting motor threshold. Stimulation is paired with an fMRI-naming task on the first and last days of treatment. After the treatment phase is complete, the language battery obtained at baseline is repeated two and six months following stimulation in order to identify rTMS-induced changes in performance. The fMRI-naming task is also repeated two and six months following treatment. Patients who are randomized to the sham arm of the study undergo sham site-finding, sham treatment, fMRI-naming studies, and repeat language testing two months after completing sham treatment. Sham patients then cross over into the real stimulation arm, completing real site-finding, real treatment, fMRI, and two- and six-month post-stimulation language testing.

We explore the use of repetitive transcranial magnetic stimulation (rTMS) to improve language abilities in patients with chronic stroke and non-fluent aphasia. After identifying a site in the right frontal gyrus for each patient that responds optimally to stimulation, we target this site during ten days of rTMS treatment.

Transcranial magnetic stimulation (TMS) has been shown to significantly improve language function in patients with non-fluent aphasia1. In this ...

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

Modeling and Simulations of Olfactory Drug Delivery with Passive and Active Controls of Nasally Inhaled Pharmaceutical Aerosols

There are many advantages of direct nose-to-brain drug delivery in the treatment of neurological disorders. However, its application is limited by the extremely low delivery efficiency (&#60; 1%) to the olfactory mucosa that directly connects the brain. It is crucial to develop novel techniques to deliver neurological medications more effectively to the olfactory region. The objective of this study is to develop a numerical platform to simulate and improve intranasal olfactory drug delivery. A coupled image-CFD method was presented that synthetized the image-based model development, quality meshing, fluid simulation, and magnetic particle tracking. With this method, performances of three intranasal delivery protocols were numerically assessed and compared. Influences of breathing maneuvers, magnet layout, magnetic field strength, drug release position, and particle size on the olfactory dosage were also numerically studied.
From the simulations, we found that clinically significant olfactory dosage (up to 45%) were feasible using the combination of magnet layout and selective drug release. A 64 -fold higher delivery of dosage was predicted in the case with magnetophoretic guidance compared to the case without it. However, precise guidance of nasally inhaled aerosols to the olfactory region remains challenging due to the unstable nature of magnetophoresis, as well as the high sensitivity of olfactory dosage to patient-, device-, and particle-related factors.

This manuscript reviews the modeling and simulations of different protocols to deliver medications to the olfactory region in image-based nasal airway models. Multiple software modules are used to develop the anatomically accurate nose model, generate computational mesh, simulate nasal airflows, and predict particle deposition at the olfactory region.

There are many advantages of direct nose-to-brain drug delivery in the treatment of neurological disorders. However, its application is limited by the ...

Mechanical Ventilation Boot Camp Curriculum

Medical management of mechanically ventilated patients is challenging to novice providers. Incorrect management of this population may lead to increased morbidity and mortality. A three-day simulation-based boot camp serves to provide one-on-one instruction with a critical care provider. These intensivists may dispense personalized immediate feedback as learners engage in hands-on practice with a real mechanical ventilator. Multiple different pathologies can be reviewed that may not be encountered in the clinical setting. Learners can visualize immediate consequences of their actions and may troubleshoot and ask questions, all while in a safe learning environment. We describe the use of human-patient simulators connected to breathing simulators and mechanical ventilators. Potential curriculum executors should be aware of the cost of the equipment and the time needed to dedicate to boot camp execution; however, this intensive interactive training has been shown to increase provider competency, knowledge, and confidence in ventilator management. This curriculum outline provides guidance on how to execute a simulation-based boot camp to train providers on the management of mechanically ventilated patients.

This curriculum outlines how to execute a simulation-based boot camp to teach providers how to manage mechanically ventilated patients.

Medical management of mechanically ventilated patients is challenging to novice providers. Incorrect management of this population may lead to increased ...

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to injury. In fact, the most common type of eye injury found in clinic is a corneal abrasion. Here, we utilize an ocular burr to induce an abrasion resulting in removal of the corneal epithelium in vivo on anesthetized mice. This method allows for targeted and reproducible epithelial disruption, leaving other areas intact. In addition, we describe the visualization of the abraded epithelium with fluorescein staining and provide concrete advice on how to visualize the abraded cornea. Then, we follow the timeline of wound healing 0, 18, and 72 h after abrasion, until the wound is re-epithelialized. The epithelial abrasion model of corneal injury is ideal for studies on epithelial cell proliferation, migration and re-epithelialization of the corneal layers. However, this method is not optimal to study stromal activation during wound healing, because the ocular burr does not penetrate to the stromal cell layers. This method is also suitable for clinical applications, for example, pre-clinical test of drug effectiveness.

This protocol describes a method to inflict an abrasion to the ocular surface of the mouse, and to follow the wound healing process thereafter. The protocol takes advantage of an ocular burr to partially remove the surface epithelium of the eye in anaesthetized mice.

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...

A Case Series of Successful Abdominal Closure Utilizing a Novel Technique Combining a Mechanical Closure System with a Biologic Xenograft that Accelerates Wound Healing

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Utilizing Repetitive Transcranial Magnetic Stimulation to Improve Language Function in Stroke Patients with Chronic Non-fluent Aphasia

Murine Model of Wound Healing

Modeling and Simulations of Olfactory Drug Delivery with Passive and Active Controls of Nasally Inhaled Pharmaceutical Aerosols

Mechanical Ventilation Boot Camp Curriculum

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing