Name: a detailed protocol for perspiration monitoring using a novel, small, wireless device
Uploaded: 2016-11-24T15:00:00
Description: Watch this Scientific Journal Video about A Detailed Protocol for Perspiration Monitoring Using a Novel, Small, Wireless Device at JoVE.com

We would like to acknowledge Mr. Ryohei Suganuma and Ms. Sakie Tachibana who helped perform the research. This study was supported in part by the JSPS KAKENHI Grant Numbers 15K20664, 24500848, and 21500405. This study was also funded in part by the MEXT/JST Tenure Track Promotion Program. A part of this study was based on the Japanese Unexamined Patent Application Publication No. 2011-169881 and the Japanese Patent No. 5708911.

NOTE: The device, including the method of analysis, is covered by Japanese Unexamined Patent Application Publication No. 2011-169881 and the Japanese Patent No. 5708911. This study, including the protocol of the experiment with human subjects, was approved by the Medical Ethics Committee of Kanazawa University (#553-1).
1. Prerequisites for the Perspiration Monitoring Device
NOTE: Perform these steps only once before the first use.
<ol>
	<li>Install the universal ...

<ol>
	<li>Hardy, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiology+of+temperature+regulation.">Physiology of temperature regulation.</a> Physiol. Rev. 41, 521-606 (1961).</li><li>Niepomniszcze, H., Amad, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Skin+disorders+and+thyroid+diseases.">Skin disorders and thyroid diseases.</a> J. Endocrinol. Invest. 24 (8), 628-638 (2001).</li><li>Korpelainen, J. T., Sotaniemi, K. A., Myllyla, V. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperhidrosis+as+a+reflection+of+autonomic+failure+in+patients+with+acute+hemispheral+brain+infarction.+An+evaporimetric+study.">Hyperhidrosis as a reflection of autonomic failure in patients with acute hemispheral brain infarction. An evaporimetric study.</a> Stroke. 23 (9), 1271-1275 (1992).</li><li>Fealey, R. D., Low, P. A., Thomas, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermoregulatory+sweating+abnormalities+in+diabetes+mellitus.">Thermoregulatory sweating abnormalities in diabetes mellitus.</a> Mayo Clin Proc. 64 (6), 617-628 (1989).</li><li>Leung, A. K., Chan, P. Y., Choi, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperhidrosis.">Hyperhidrosis.</a> Int J Dermatol. 38 (8), 561-567 (1999).</li><li>Kronenberg, F., Cote, L. J., Linkie, D. M., Dyrenfurth, I., Downey, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Menopausal+hot+flashes:+thermoregulatory,+cardiovascular,+and+circulating+catecholamine+and+LH+changes.">Menopausal hot flashes: thermoregulatory, cardiovascular, and circulating catecholamine and LH changes.</a> Maturitas. 6 (1), 31-43 (1984).</li><li>Gibson, L. E., Cooke, R. 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Eng. 62 (6), 1457-1465 (2015).</li><li>Gao, 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=Fully+integrated+wearable+sensor+arrays+for+multiplexed+in+situ+perspiration+analysis.">Fully integrated wearable sensor arrays for multiplexed in situ perspiration analysis.</a> Nature. 529 (7587), 509-514 (2016).</li><li>Nilsson, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+water+exchange+through+skin.">Measurement of water exchange through skin.</a> Med. Biol. Eng. 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The aim of this article is to introduce the use of a novel, wireless perspiration monitoring device. Owing to the recent progress of engineering, more accurate, easy-to-handle methods for temporal perspiration monitoring have been proposed; the ventilated chamber method24,25 and the vapor pressure diffusion method23 are representative examples. However, the ventilated chamber method requires the use of dry nitrogen or a pump with desiccant to produce a dry atmosphere, and thus, the exterior tends to...

Human perspiration, generally known as &#34;sweating,&#34; is not just a mechanism for thermoregulation1, but it is also related to certain kinds of diseases. The etiology of abnormal perspiration is broad, including: heatstroke, hyper- or hypothyroidism2, brain infarction3, diabetes mellitus4, dysautonomia5, menopause (known as &#34;hot flash&#34;)6, cystic fibrosis7, Parkinson&#39;s disease8, and social anxiety disorder9. In light of the number of perspiration-related diseases, it has been considered beneficial to monitor perspiration rates for the early diagnosis or prediction of such diseases (e.g., prevention of heatstroke) in a ubiquitous manner10.
To date, only a small number of devices for perspiration monitoring have been proposed. In early days, skin conductance and relative humidity were used for indirect indices of the amount of perspiration11,12. Most recently, several kinds of flexible, wearable sensors for perspiration monitoring have been proposed13-19, although they are intended for the analysis of sweat electrolytes rather than the amount or temporal pattern of perspiration. The calculation of water vapor diffusion has been utilized for a more quantitative method of monitoring water exchange from the skin20-23. However, this requires (1) the assumption that the outer atmosphere is still and constant20, (2) enough sensitivity to detect the natural flow of water vapor21,22, or (3) a coolant (e.g., Peltier device that consumes a substantial amount of electricity) to condense water vapor to liquid23; thus, they might be difficult for daily and long-term monitoring. As an alternative, a ventilated chamber method was developed20,24,25. In the ventilated chamber method, dry nitrogen or dehumidified air is infiltrated in a small chamber adjacent to the skin from a nitrogen gas tank or a pump, and gas in the water vapor evaporated from the skin is collected. The amount of water vapor from the skin can be calculated from the difference of the humidity in the outlet and inlet gases. Although this method can estimate the amount of perspiration very precisely, a nitrogen gas tank or a mechanical pump is generally large enough to impede daily monitoring.
To address these drawbacks, we have recently developed a novel device for perspiration monitoring, in which a closed chamber with a desiccant-driven enforced water vapor flow, enabled sensitive and long-term monitoring26. This device consists of a cylindrical plastic exterior, temperature/relative humidity (T/RH) sensor with recording microprocessor, and silica gel (Figure 1). In principle, the outer atmosphere should not interfere with the water vapor flow, and a coolant or ventilating chamber is not required. Perspiration profiles can be obtained by solving equations using a spreadsheet software26. A previous study has only shown the principle of the developed device and has omitted the detailed method for how to use the device because of space limitations.
The objective of this article, therefore, is to show a detailed method of how to use the developed device for perspiration monitoring, by showing the recording of stress-induced palmar perspiration during the sympathetic activity test as an example.

<table border="0" cellpadding="0" cellspacing="0" width="1173">
	<tbody>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:1173px;">Required for perspiration monitoring</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Perspiration monitoring device</td>
			<td style="width:264px;">Rousette Strategy Inc.</td>
			<td style="width:127px;">SNT-200</td>
			<td style="width:465px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">USB-serial port conversion interface</td>
			<td style="width:264px;">Rousette Strategy Inc.</td>
			<td style="width:127px;">UUI-200</td>
			<td style="width:465px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Perspiration recording software</td>
			<td style="width:264px;">Rousette Strategy Inc.</td>
			<td style="width:127px;">TED99</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Silica gel</td>
			<td style="width:264px;">Wako Pure Chemical Industries Ltd.</td>
			<td style="width:127px;">194-16665</td>
			<td>Type A silica gel should be used.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Medical double-sided tape</td>
			<td style="width:264px;">3M</td>
			<td style="width:127px;">2181</td>
			<td>Medical grade is recommended because of the attachment to the skin.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Computer</td>
			<td style="width:264px;"></td>
			<td style="width:127px;"></td>
			<td>Requires Windows operating system.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Name</td>
			<td style="width:264px;">Company</td>
			<td style="width:127px;">Catalog</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:1173px;">Required for the monitoring of sympathetic skin response</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Instrumentation amplifier</td>
			<td style="width:264px;">Nihon Kohden Corp.</td>
			<td style="width:127px;">AB-611J</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Amplifier chassis</td>
			<td style="width:264px;">Nihon Kohden Corp.</td>
			<td style="width:127px;">MEG-6108</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Input box</td>
			<td style="width:264px;">Nihon Kohden Corp.</td>
			<td style="width:127px;">JB-610B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Alcohol swab</td>
			<td style="width:264px;">Suzuran Sanitary Goods Co., Ltd.</td>
			<td style="width:127px;">4545766050846</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Electrodes</td>
			<td style="width:264px;">Nihon Kohden Corp.</td>
			<td style="width:127px;">NE-114A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Electrode paste</td>
			<td style="width:264px;">Nihon Kohden Corp.</td>
			<td style="width:127px;">Z-401CE</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Medical tape</td>
			<td style="width:264px;">Nichiban Co., Ltd.</td>
			<td style="width:127px;">SG257</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Analog signal interface</td>
			<td style="width:264px;">Micro Science K.K.</td>
			<td style="width:127px;">C BOX-014</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Analog-to-digital converter</td>
			<td style="width:264px;">Micro Science K.K.</td>
			<td style="width:127px;">ADM-686PCI</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">SSR recording software</td>
			<td style="width:264px;">Matsuyama Advance Co., Ltd.</td>
			<td style="width:127px;">LaBDAQ2000</td>
			<td></td>
		</tr>
	</tbody>
</table>

a detailed protocol for perspiration monitoring using a novel, small, wireless device

Perspiration monitoring can be utilized for the detection of certain diseases, such as thermoregulation and mental disorders, particularly when the patients are unaware of such disorders or are having difficulty expressing their symptoms. Until now, several devices for perspiration monitoring have been developed; however, such devices tend to have a relatively large exterior, considerable power consumption, and/or less sensitivity.
Recently, we developed a small, wireless device for perspiration monitoring. The device consists of a temperature/relative humidity (T/RH) sensor, battery-driven small data logger, and silica gel as a desiccant in a small cylindrical exterior. The T/RH sensor is placed between the detection windows (through which the water vapor from the skin enters) and the silica gel. The underlying principle of the perspiration monitoring device is based on Fick&#39;s law of diffusion, which means that water vapor flux from the skin to the silica gel (i.e.&#160;transepidermal water loss and perspiration) can be captured by change in humidity at the T/RH sensor. In addition, a baseline subtraction method was adopted to distinguish perspiration and transepidermal water loss.
As shown in the previous report, the developed device can monitor the perspiration at any sites of the body in an easy, wireless manner. However, detailed methods of how to use the device have not been disclosed yet. In this article, therefore, we would like to show the point-by-point tutorials of how to use the device for perspiration monitoring, by showing the sympathetic activity test with the sympathetic skin response monitoring as an example.

Using this novel device for perspiration monitoring (Figure 1) and the Fick&#39;s law-based calculation, the temporal perspiration profiles can be obtained in an easy, wireless manner. Figure 2 shows representative data of wireless perspiration monitoring during the sympathetic activity test. In the experiment, the device for the perspiration monitoring, along with the electrodes for the sympathetic skin response (SSR) monitoring, was attached to the subj...

Watch this Scientific Journal Video about A Detailed Protocol for Perspiration Monitoring Using a Novel, Small, Wireless Device at JoVE.com

A Detailed Protocol for Perspiration Monitoring Using a Novel, Small, Wireless Device

Recently, we developed a small wireless device for perspiration monitoring. In this article, we present detailed protocols on how to use the device for perspiration monitoring with an example of the sympathetic activity test.

Perspiration monitoring can be utilized for the detection of certain diseases, such as thermoregulation and mental disorders, particularly when the ...

The overall goal of this procedure is to monitor perspiration patterns with a newly developed wireless device. The main advantage of this technique is that both the amount and temporal pattern of perspiration can be monitored by just attaching the device to the skin. Since this is a new device, visual demonstration of this method is critical as the preparation and usage of the device are difficult to learn.First, check the perspiration monitoring device to ensure that the battery has not been inserted. Then, plug the white connector of the USB serial port conversion interface to the receptacle at the at the top of the perspiration monitoring device. Next, execute the perspiration recording software.Open the Settings tab and set the COM number to the ID number of the serial port. Click the Confirm Connection button and check if the message, Connected, appears. Within the Data File Folder settings, choose the drive and select the folder where the perspiration data will be saved.Set the Number of Holes to four. Open the Measure and Rec tab and set the Time Interval to the desired sample time. Then, insert a battery into the perspiration monitoring device.Fill the device with dry silica gel and close the lid. Finally, click the Start Logging Now button in the perspiration recording software. Begin by cleansing the skin at the recording location with an alcohol swab.Use electrode paste to place the anode, cathode, and grounding electrode on the palm, back of the hand, and wrist respectively. Fix the electrodes with medical tape. On the instrumentation amplifier, turn the Sensitivity knob to one millivolt per volt, the Lo Cut filter to 0.5 hertz, and the Hi Cut filter to three kilohertz.Finally, execute the sympathetic skin response, or SSR recording software, and click the Start Measurement button to begin the recording at the sample rate of 200 hertz. Begin by placing medical double sided tape on the bottom of the perspiration monitoring device without obstructing the four holes at the bottom of the device. Disconnect the perspiration monitoring device from the USB serial port conversation interface and immediately ensure that the perspiration monitoring begins automatically by checking if the LED lamp is blinking.Next, remove the release liner of the medical double sided tape and put the perspiration monitoring device on the skin where perspiration is to be monitored. Wait 10 minutes for stabilization of water vapor diffusion, then start the sympathetic activity test. After the examination, remove the perspiration monitoring device from the skin.Click the Stop Measurement button in the SSR recording software to stop the SSR recording and remove all electrodes from the skin. To perform the perspiration analysis, first connect the perspiration monitoring device to the USB serial port conversion interface. Then, open the perspiration monitoring software, click the Settings tab, and click the Confirm Connection button.Then, open the Measure and Rec tab and click the Download button to save the raw data of the device in the specified folder. Finally, select the Analysis tab and click Read Data File. Observe the open file dialog popup.Select the saved file and then click the Open button to allow the software to complete the analysis. As a result of the sympathetic activity test, palmar perspiration along with the SSR reaction was observed in response to the sympathetic activities, such as taking a deep inspiration or doing a mental calculation. Furthermore, a one hour recording of perspiration at the palm and the anterior chest showed distinct patterns according to different activities.After watching this video you should have a good understanding of how to use this novel wireless device for measuring the perspiration.

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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. ...

A Novel Technique of Rescuing Capsulorhexis Radial Tear-out using a Cystotome

Part 1 : Purpose: To demonstrate a capsulorhexis radial tear out rescue technique using a cystotome on a virtual reality cataract surgery simulator and in a human eye. Part 2 : Method: Steps: When a capsulorhexis begins to veer radially towards the periphery beyond the pupillary margin the following steps should be applied without delay. 2.1) Stop further capsulorhexis manoeuvre and reassess the situation. 2.2) Fill the anterior chamber with ophthalmic viscosurgical device (OVD). We recommend mounting the cystotome to a syringe containing OVD so that the anterior chamber can be reinflated rapidly. 2.3) The capsulorhexis flap is then left unfolded on the lens surface. 2.4) The cystotome tip is tilted horizontally to avoid cutting or puncturing the flap and is engaged on the flap near the leading edge of the tear but not too close to the point of tear. 2.5) Gently push or pull the leading edge of tear opposite to the direction of tear. 2.6) The leading tearing edge will start to do a 'U-Turn'. Maintain the tension on the flap until the tearing edge returns to the desired trajectory. Part 3 : Results: Using our technique, a surgeon can respond instantly to radial tear out without having to change surgical instruments. Changing surgical instruments at this critical stage runs a risk of further radial tear due to sudden shallowing of anterior chamber as a result of forward pressure from the vitreous. Our technique also has the advantage of reducing corneal wound distortion and subsequent anterior chamber collapse. Part 4 : Discussion The EYESI Surgical Simulator is a realistic training platform for surgeons to practice complex capsulorhexis tear-out techniques. Capsulorhexis is the most important and complex part of phacoemulsification and endocapsular intraocular lens implantation procedure. A successful cataract surgery depends on achieving a good capsulorhexis. During capsulorhexis, surgeons may face a challenging situation like a capsulorhexis radial tear-out. A surgeon must learn to tackle the problem promptly without making the situation worse. Some other methods of rescuing the situation have been described using a capsulorhexis forceps. However, we believe our method is quicker, more effective and easier to manipulate as demonstrated on the EYESi surgical simulator and on a human eye. Acknowledgments: List acknowledgements and funding sources. We would like to thank Dr. Wael El Gendy, for video clip. Disclosures: describe potential conflicting interests or state We have nothing to disclose. References: 1. Brian C. Little, Jennifer H. Smith, Mark Packer. J Cataract Refract Surg 2006; 32:1420 1422, Issue-9. 2. Neuhann T. Theorie und Operationstechnik der Kapsulorhexis. Klin Monatsbl Augenheilkd. 1987; 1990: 542-545. 3. Gimbel HV, Neuhann T. Development, advantages and methods of the continuous circular capsulorhexis technique. J Cataract Refract Surg. 1990; 16: 31-37. 4. Gimbel HV, Neuhann T. Continuous curvilinear capsulorhexis. (letter) J Cataract Refract Sur. 1991; 17: 110-111.

Capsulorhexis is an important step in phacoemulsification surgery. A surgeon creates a continous curvilinear tear on the anterior lens capsule by controlling the tearing vector forces. A peripherally extended tear is a serious complication. This video demonstrates a novel technique of rescuing capsular radial tear out using a cystotome.

Part 1 : Purpose:   To demonstrate a capsulorhexis radial tear out rescue technique using a cystotome on a virtual reality cataract surgery simulator and ...

The Trier Social Stress Test Protocol for Inducing Psychological Stress

This article demonstrates a psychological stress protocol for use in a laboratory setting. Protocols that allow researchers to study the biological pathways of the stress response in health and disease are fundamental to the progress of research in stress and anxiety.1 Although numerous protocols exist for inducing stress response in the laboratory, many neglect to provide a naturalistic context or to incorporate aspects of social and psychological stress. Of psychological stress protocols, meta-analysis suggests that the Trier Social Stress Test (TSST) is the most useful and appropriate standardized protocol for studies of stress hormone reactivity.2 In the original description of the TSST, researchers sought to design and evaluate a procedure capable of inducing a reliable stress response in the majority of healthy volunteers.3 These researchers found elevations in heart rate, blood pressure and several endocrine stress markers in response to the TSST (a psychological stressor) compared to a saline injection (a physical stressor).3 Although the TSST has been modified to meet the needs of various research groups, it generally consists of a waiting period upon arrival, anticipatory speech preparation, speech performance, and verbal arithmetic performance periods, followed by one or more recovery periods. The TSST requires participants to prepare and deliver a speech, and verbally respond to a challenging arithmetic problem in the presence of a socially evaluative audience.3 Social evaluation and uncontrollability have been identified as key components of stress induction by the TSST.4 In use for over a decade, the goal of the TSST is to systematically induce a stress response in order to measure differences in reactivity, anxiety and activation of the hypothalamic-pituitary-adrenal (HPA) or sympathetic-adrenal-medullary (SAM) axis during the task.1 Researchers generally assess changes in self-reported anxiety, physiological measures (e.g. heart rate), and/or neuroendocrine indices (e.g. the stress hormone cortisol) in response to the TSST. Many investigators have adopted salivary sampling for stress markers such as cortisol and alpha-amylase (a marker of autonomic nervous system activation) as an alternative to blood sampling to reduce the confounding stress of blood-collection techniques. In addition to changes experienced by an individual completing the TSST, researchers can compare changes between different treatment groups (e.g. clinical versus healthy control samples) or the effectiveness of stress-reducing interventions.1

This article describes a protocol for inducing psychological stress in participants, which enables researchers to measure psychological, physiological and neuroendocrine responses to stress within single participants or between groups.

This article demonstrates a psychological stress protocol for use in a laboratory setting.  Protocols that allow researchers to study the biological ...

Protocol for Long Duration Whole Body Hyperthermia in Mice

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body temperature that is observed during fever. The use of hyperthermia as an adjuvant has emerged as a promising procedure for tumor regression in the field of cancer biology. For this purpose, the most important requirement is to have reliable and uniform heating protocols. We have developed a protocol for hyperthermia (whole body) in mice. In this protocol, animals are exposed to cycles of hyperthermia for 90 min followed by a rest period of 15 min. During this period mice have easy access to food and water. High body temperature spikes in the mice during first few hyperthermia exposure cycles are prevented by immobilizing the animal. Additionally, normal saline is administered in first few cycles to minimize the effects of dehydration. This protocol can simulate fever like conditions in mice up to 12-24 hr. We have used 8-12 weeks old BALB/Cj female mice to demonstrate the protocol.

This paper describes a protocol for whole body hyperthermia in mice that can stimulate fever like conditions up to 12-24 hr.

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body ...

Collection Protocol for Human Pancreas

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation of pancreas recovered from cadaveric organ donors. The pancreas is divided into 3 main regions (head, body, tail) followed by serial transverse sections throughout the medial to lateral axis. Alternating sections are used for fixed paraffin and fresh frozen blocks and remnant samples are minced for snap frozen sample preparations, either with or without RNAse inhibitors, for DNA, RNA, or protein isolation. The overall goal of the pancreas dissection procedure is to sample the entire pancreas while maintaining anatomical orientation.
Endocrine cell heterogeneity in terms of islet composition, size, and numbers is reported for human islets compared to rodent islets 1. The majority of human islets from the pancreas head, body and tail regions are composed of insulin-containing &beta;-cells followed by lower proportions of glucagon-containing &alpha;-cells and somatostatin-containing &delta;-cells. Pancreatic polypeptide-containing PP cells and ghrelin-containing epsilon cells are also present but in small numbers. In contrast, the uncinate region contains islets that are primarily composed of pancreatic polypeptide-containing PP cells 2. These regional islet variations arise from developmental differences. The pancreas develops from the ventral and dorsal pancreatic buds in the foregut and after rotation of the stomach and duodenum, the ventral lobe moves and fuses with the dorsal 3. The ventral lobe forms the posterior portion of the head including the uncinate process while the dorsal lobe gives rise to the rest of the organ. Regional pancreatic variation is also reported with the tail region having higher islet density compared to other regions and the dorsal lobe-derived components undergoing selective atrophy in type 1 diabetes4,5.
Additional organs and tissues are often recovered from the organ donors and include pancreatic lymph nodes, spleen and non-pancreatic lymph nodes. These samples are recovered with similar formats as for the pancreas with the addition of isolation of cryopreserved cells. When the proximal duodenum is included with the pancreas, duodenal mucosa may be collected for paraffin and frozen blocks and minced snap frozen preparations.

This video demonstrates a dissection procedure for processing human pancreas into multiple storage formats. Anatomical orientation is maintained throughout the pancreatic regions to allow definition of regional islet composition and density.

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation ...

Parabiosis in Mice: A Detailed Protocol

Parabiosis is a surgical union of two organisms allowing sharing of the blood circulation. Attaching the skin of two animals promotes formation of microvasculature at the site of inflammation. Parabiotic partners share their circulating antigens and thus are free of adverse immune reaction. First described by Paul Bert in 18641, the parabiosis surgery was refined by Bunster and Meyer in 1933 to improve animal survival2. In the current protocol, two mice are surgically joined following a modification of the Bunster and Meyer technique. Animals are connected through the elbow and knee joints followed by attachment of the skin allowing firm support that prevents strain on the sutured skin. Herein, we describe in detail the parabiotic joining of a ubiquitous GFP expressing mouse to a wild type (WT) mouse. Two weeks after the procedure, the pair is separated and GFP positive cells can be detected by flow cytometric analysis in the blood circulation of the WT mouse. The blood chimerism allows one to examine the contribution of the circulating cells from one animal in the other.

Parabiotic joining of two organisms leads to the development of a shared circulatory system. In this protocol, we describe the surgical steps to form a parabiotic connection between a wild-type mouse and a constitutive GFP-expressing mouse.

Parabiosis is a surgical union of two organisms allowing sharing of the blood circulation. Attaching the skin of two animals promotes formation of ...

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

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

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

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

A Detailed Protocol for Physiological Parameters Acquisition and Analysis in Neurosurgical Critical Patients

Intracranial pressure (ICP) monitoring is now widely used in neurosurgical critical patients. Besides mean ICP value, the ICP derived parameters such as ICP waveform, amplitude of pulse (AMP), the correlation of ICP amplitude and ICP mean (RAP), pressure reactivity index (PRx), ICP and arterial blood pressure (ABP) wave amplitude correlation (IAAC), and so on, can reflect intracranial status, predict prognosis, and can also be used as guidance of proper treatment. However, most of the clinicians focus only on the mean ICP value while ignoring these parameters because of the limitations of the current devices. We have recently developed a multimodality monitoring system to address these drawbacks. This portable, user-friendly system will use a data collecting and storing device to continuously acquire patients&#39; physiological parameters first, i.e., ABP, ICP, and oxygen saturation, and then analyze these physiological parameters. We hope that the multimodality monitoring system will be accepted as a key measure to monitor physiological parameters, to analyze the current clinical status, and to predict the prognosis of the neurosurgical critical patients.

Recently, we have developed a portable multimodality monitoring system for the monitoring of various physiological parameters in neurosurgical critical patients. Detailed protocols on how to use this multimodality monitoring system are presented here.

Intracranial pressure (ICP) monitoring is now widely used in neurosurgical critical patients. Besides mean ICP value, the ICP derived parameters such as ...

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The amount of the different tissues in the callus provides important information on the fracture healing progress. Available in vivo techniques to longitudinally monitor the callus tissue development in preclinical fracture-healing studies using small animals include digital radiography and &#181;CT imaging. However, both techniques are only able to distinguish between mineralized and non-mineralized tissue. Consequently, it is impossible to discriminate cartilage from fibrous tissue. In contrast, magnetic resonance imaging (MRI) visualizes anatomical structures based on their water content and might therefore be able to noninvasively identify soft tissue and cartilage in the fracture callus. Here, we report the use of an MRI-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice. The experiments demonstrated that the fixator and a custom-made mounting device allow repetitive MRI scans, thus enabling longitudinal analysis of fracture-callus tissue development.

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

The evaluation of tissue development in the fracture callus during endochondral bone healing is essential to monitor the healing process. Here, we report the use of a magnetic resonance imaging (MRI)-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice.

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The ...