This research was partially supported by the Japan Agency for Medical Research and Development (AMED) under the Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS) project (Grant Number JP23dm0207001).

The present study was performed with the approval (W2023-2-041) of the Animal Experiments Committee of RIKEN (Saitama, Japan) and was conducted in accordance with the Institutional Guidelines for Experiments using Animals. In the present study, six female marmosets (2-6 years old) were used.
1. Criteria for commencement of treatment
<ol>
	<li>Start treatment when any one of the following criteria is observed: i) body weight &#60; 300 g or a decrease of &#62;20 g in the body weight per month; ii) hypoalbuminemia (serum albumin &#60; 3.8 g/dL); iii) anemia (hematocrit &#60; 35%); iv) chronic diarrhea without Clostridium difficile, enteropathogenic Escherichia coli, or intestinal protozoa; and v) an apparent decrease in muscle mass. 
	NOTE: Because of dehydration and subsequent hemoconcentration accompanied by WMS, the values of serum albumin and hematocrit may be artificially elevated in affected animals.</li>
</ol>
2. Oral administration
<ol>
	<li>Mix 0.1 mL of the iron formulation (see the Table of Materials) with a piece of sponge cake.</li>
	<li>Give the mash to the treated marmoset. 
	NOTE: Ensure that the treated marmoset has completely swallowed the mash.</li>
</ol>
3. Subcutaneous administration
<ol>
	<li>Catch the WMS-affected marmoset in the home cage by holding the tail gently.</li>
	<li>Hold the animal&#39;s upper body from the back with the other hand.</li>
	<li>Bring the animal to the treatment room.</li>
	<li>Let the animal cling to the assistant&#39;s chest or arm.</li>
	<li>Clean the area of the animal&#39;s back with an alcohol swab.</li>
	<li>Inject 0.1 mL of tranexamic acid (5% solution, see the Table of Materials) into the back subcutaneously using a 26 G needle 5x per week.</li>
	<li>Inject 5.0 mL of Ringer&#39;s lactate (see the Table of Materials) with 0.1 mL of a vitamin formulation (see the Table of Materials) subcutaneously using a 26 G needle 3x per week. 
	NOTE: Each injection should be done at each different spot within 1 min.</li>
</ol>
4. Intravenous administration of amino acid formulation
<ol>
	<li>Catch the WMS-affected marmoset in the home cage by holding the tail gently.</li>
	<li>Hold the animal&#39;s upper body from the back with the other hand.</li>
	<li>Bring the animal to the treatment room.</li>
	<li>Have the assistant restrain the animal&#39;s upper body by holding the base of both arms and restrain the lower body by holding the base of both legs with the other hand.Ask the assistant to place the thumb restraining the animal&#39;s lower body on the anterior side of the animal&#39;s knee to prevent it from bending.</li>
	<li>Clean the animal&#39;s calf with an alcohol swab.</li>
	<li>Inject 2.0 mL of the amino acid formulation (see the Table of Materials) intravenously via the saphenous vein using a 27 G butterfly needle three times per week. 
	NOTE: Injection must be done as slowly as possible up to 2 min.</li>
	<li>After injection, ask the assistant to press the area with absorbent cotton until bleeding stops.</li>
</ol>
5. Body weight measurement
<ol>
	<li>After treatment (intravenous and/or subcutaneous injection), weigh the animal with a carrying box once a week to monitor the body weight. 
	NOTE: Weigh the marmoset at around the same time because the body weight is affected by the presence of food.</li>
</ol>
6. Complete blood count (CBC) analysis and serum chemistry tests
<ol>
	<li>Catch the WMS-affected marmoset in the home cage by holding the tail gently.</li>
	<li>Hold the animal&#39;s upper body from the back with the other hand.</li>
	<li>Bring the animal to the treatment room.</li>
	<li>Have the assistant restrain the animal&#39;s upper body by holding the base of both arms and restrain the lower body by holding the base of either leg with the other hand.</li>
	<li>Ask the practitioner to hold the animal&#39;s leg that is not restrained.</li>
	<li>Clean the animal&#39;s inguinal region with an alcohol swab.</li>
	<li>Collect 0.4 mL of blood from the femoral vein using a 26 G needle monthly.</li>
	<li>Use 0.05 mL of the blood sample for CBC analysis, including hematocrit.</li>
	<li>Let the rest of the blood sample stand for 1 h at room temperature and centrifuge at 1,800 &#215; g for 20 min.</li>
	<li>Use the serum for chemistry tests, including albumin. 
	NOTE: The volume of the blood sample can be reduced by minimizing the number of parameters for the serum chemistry test. Adjust the volume of the blood sample according to the individual marmoset&#39;s condition.</li>
</ol>

An Efficient Modified Method for Treating Wasting Marmoset Syndrome

<ol>
	<li>Baxter, V. 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=Serum+albumin+and+body+weight+as+biomarkers+for+the+antemortem+identification+of+bone+and+gastrointestinal+disease+in+the+common+marmoset.">Serum albumin and body weight as biomarkers for the antemortem identification of bone and gastrointestinal disease in the common marmoset.</a> PLoS ONE. 8 (12), e82747 (2013).</li><li>Brack, M., Rothe, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+tubulointerstitial+nephritis+and+wasting+disease+in+marmosets+(Callithrix+jacchus).">Chronic tubulointerstitial nephritis and wasting disease in marmosets (Callithrix jacchus).</a> Vet Pathol. 18, 45-54 (1981).</li><li>Chalifoux, L. V., Bronson, R. T., Escajadillo, A., McKenna, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+analysis+of+the+association+of+gastroenteric+lesions+with+chronic+wasting+syndrome+of+marmosets.">An analysis of the association of gastroenteric lesions with chronic wasting syndrome of marmosets.</a> Vet Pathol Suppl. 7, 141-162 (1982).</li><li>Logan, A. C., Khan, K. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+pathologic+changes+in+two+marmosets+with+wasting+syndrome.">Clinical pathologic changes in two marmosets with wasting syndrome.</a> Toxicol Pathol. 24 (6), 707-709 (1996).</li><li>Ludlage, E., Mansfield, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+care+and+diseases+of+the+common+marmoset+(Callithrix+jacchus).">Clinical care and diseases of the common marmoset (Callithrix jacchus).</a> Comp Med. 53 (4), 369-382 (2003).</li><li>Ross, C. N., Davis, K., Dobek, G., Tardif, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aging+phenotypes+of+common+marmosets+(Callithrix+jacchus).">Aging phenotypes of common marmosets (Callithrix jacchus).</a> J Aging Res. 2012, 567143 (2012).</li><li>Yoshimoto, T., Niimi, K., Takahashi, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Curative+effects+of+tranexamic+acid+with+supportive+measures+on+wasting+marmoset+syndrome.">Curative effects of tranexamic acid with supportive measures on wasting marmoset syndrome.</a> Comp Med. 66 (6), 1-6 (2016).</li><li>Niimi, 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=Measurement+of+the+&#945;1-proteinase+inhibitor+(&#945;1-antitrypsin)+of+common+marmoset+and+intestinal+protein+loss+in+wasting+syndrome.">Measurement of the &#945;1-proteinase inhibitor (&#945;1-antitrypsin) of common marmoset and intestinal protein loss in wasting syndrome.</a> Biosci Rep. 39 (7), 20190562 (2019).</li><li>Otovic, P., Smith, S., Hutchinson, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+glucocorticoids+in+marmoset+wasting+syndrome.">The use of glucocorticoids in marmoset wasting syndrome.</a> J Med Primatol. 44 (2), 53-59 (2015).</li><li>Turner, P. V., Brabb, T., Pekow, C., Vasbinder, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Administration+of+substances+to+laboratory+animals:+routes+of+administration+and+factors+to+consider.">Administration of substances to laboratory animals: routes of administration and factors to consider.</a> J Am Assoc Lab Anim Sci. 50 (5), 600-613 (2011).</li><li>Lewis, S. M., Hotchkiss, C. E., Ullrey, D. E., Wolfe-Coote, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+13.+Nutrition+and+nutritional+diseases.">Chapter 13. Nutrition and nutritional diseases.</a> The Laboratory Primates. , 181-208 (2005).</li></ol>

The authors have no conflicts of interest to disclose.

In 2016, we reported that tranexamic acid with supportive measures was an effective treatment for WMS, which was the first report to demonstrate WMS therapy without the use of glucocorticoids. In humans, glucocorticoids are considered the most effective treatment of IBD. However, predonizolone, one of the most commonly used glucocorticoids, is not suitable for WMS treatment because of its adverse effects. Although budesonide, a glucocorticoid, was reported for WMS treatment9, the therapy was relatively ineffective in animals with acute forms of WMS. Tranexamic acid is a plasmin inhibitor that has anti-inflammatory effects, and no notable side effects were seen in our original methods. However, although the original treatment protocol was definitely effective, it imposed a heavy load for animals and carers.
In the present study, the modified treatment methods for WMS were conducted to decrease both the animal's physiological load and the carers' workload. In the modified methods, the administration route of tranexamic acid was changed (from intraperitoneal to subcutaneous), which contributed to reducing the risk of injury to the abdominal organs. The tranexamic acid solution was not diluted in this protocol to reduce contamination risk and preparation time. In the original methods, 3.0 mL of the amino acid formulation were injected intravenously. However, the volume was somewhat large because 5.0 mL/kg is recommended to be administered as a bolus10. Therefore, the volume was reduced in the modified methods. 
The vitamin formulation used in this protocol contains vitamins B and C. In the original method, 0.5 mL of the vitamin formulation contains 2.5 mg of thiamine chloride hydrochloride, which was 2.5x the requirement of laboratory housed postweaning nonhuman primates11. In the present method, 0.1 mL of the vitamin formulation was administered, which contains 1.0 mg of thiamine chloride hydrochloride. The frequency of administration of the iron formulation and tranexamic acid solution was reduced in the modified methods, which contributed to the reduction of burden for the animals as well as the carers. 
As written in the Representative Results section, there were significant treatment effects in the modified methods. The average treatment term was 37.8 &plusmn; 25.34 days, which was shorter than that of the original methods (56 days). Since marmosets are sensitive to mental stress, prolonged treatments have the opposite effect on the animal such as reduced appetite. Thus, we recommend that the carers should comprehensively decide the timing of treatment termination based on not only the value of each parameter but also the animal's behavior.

Wasting marmoset syndrome (WMS) is a serious disease that is unique to captive common marmoset colonies. The main symptoms are weight loss, decreased muscle mass, anemia, hypoalbuminemia, and chronic enteritis1,2,3,4. Some studies have reported that 60.5% of euthanized captive marmosets had evidence of chronic enteritis5, and 31-44% of deaths involved this illness6. The high prevalence of this disease necessitated its effective treatment.
In 2016, we reported that tranexamic acid with supportive measures was an effective treatment for WMS7. In addition, intestinal protein loss of WMS-affected marmosets was significantly attenuated with this treatment8. Although the original treatment protocol was definitely effective, it included a somewhat high volume of intravenous injection, a high dose of vitamin formulation, and daily restraining. Since marmosets are susceptible to hand-capture and restraining, these actions must be less frequent. Thus, we aimed to reduce the heavy load for animals with some modification.
In the present paper, we will provide the modified methods: subcutaneous injection of undiluted tranexamic acid 5 times per week (instead of intraperitoneal injection of 5-fold-diluted tranexamic acid solution 7 times per week), 2.0 mL of amino acid formulation for intravenous injection (instead of 3.0 mL), 0.1 mL of vitamin formulation for subcutaneous injection (instead of 0.5 mL), and oral administration of iron formulation 5 times per week (instead of 7 times per week).

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1.0 mL Syringe</td>
			<td>Terumo Corporation, Tokyo, Japan</td>
			<td>SS-01T</td>
			<td>For subcutaneously injection of Tranexamic acid</td>
		</tr>
		<tr>
			<td>10 mL Syringe</td>
			<td>Terumo Corporation, Tokyo, Japan</td>
			<td>SS-10SZ</td>
			<td>For Intravenous Administration of Amino acid formulation</td>
		</tr>
		<tr>
			<td>26 G Needle</td>
			<td>Terumo Corporation, Tokyo, Japan</td>
			<td>NN-2613S</td>
			<td>For subcutaneously injection</td>
		</tr>
		<tr>
			<td>27 G Butterfly Needle Type D</td>
			<td>Terumo Corporation, Tokyo, Japan</td>
			<td>SV-27DL</td>
			<td>For Intravenous Administration of Amino acid formulation</td>
		</tr>
		<tr>
			<td>5 mL Syringe</td>
			<td>Terumo Corporation, Tokyo, Japan</td>
			<td>SS-05SZ</td>
			<td>For subcutaneously injection of Ringer&#39;s lactate and Vitamin formulation</td>
		</tr>
		<tr>
			<td>Aminoleban Injection</td>
			<td>Otsuka Pharma Factory Inc., Tokyo, Japan</td>
			<td>1942</td>
			<td>For subcutaneously injection of Ringer&#39;s lactate and Vitamin formulation</td>
		</tr>
		<tr>
			<td>C-PARA</td>
			<td>Takata Pharma Co., Ltd., Saitama, Japan</td>
			<td>120-31960-0</td>
			<td>Vitamin formulation</td>
		</tr>
		<tr>
			<td>FCV Liquid</td>
			<td>Minerva Corporation, Saitama, Japan</td>
			<td></td>
			<td>Iron formulation</td>
		</tr>
		<tr>
			<td>Lactec Injection</td>
			<td>Otsuka Pharma Factory Inc., Tokyo, Japan</td>
			<td>0714</td>
			<td>Ringer&#8217;s lactate&#160;</td>
		</tr>
		<tr>
			<td>Vasolamin Injection</td>
			<td>Meiji Seika Pharma Co., Ltd., Tokyo, Japan</td>
			<td>DBASOS</td>
			<td>Tranexamic acid</td>
		</tr>
	</tbody>
</table>

modification of the treatment methods for wasting marmoset syndrome with tranexamic acid and supportive measures

Wasting marmoset syndrome (WMS), a serious disease in captive common marmoset (Callithrix jacchus) colonies, is associated with a high mortality rate. The specific cause of WMS is still unclear and there are few effective treatments. Previously, we had reported a tranexamic acid therapy with supportive measures as a useful treatment for WMS. In the present study, we describe the modified method: a combination of 0.1 mL of 5% tranexamic acid subcutaneously five times per week, 2.0 mL of amino acid formulation intravenously three times per week, 5.0 mL of Ringer's lactate with 0.1 mL of a vitamin formulation subcutaneously three times per week, and oral administration of 0.1 mL of an iron formulation five times per week. We also describe how to administer the solution intravenously via the saphenous vein with a tip of restraining the animal, as well as the detailed methods for oral and subcutaneous administration. The modified methods have comparable efficiency to the original WMS treatment method.

The modified treatment methods resulted in increased hematocrit (Figure 1A), serum albumin (Figure 1B), and body weight (Figure 1C) in the WMS-affected marmosets. Significant differences in hematocrit (P &#60; 0.05, Wilcoxon matched-pairs signed rank test), serum albumin (P &#60; 0.05, Wilcoxon matched-pairs signed rank test), and body weight (P &#60; 0.05, Wilcoxon matched-pairs signed rank test) were seen between pre and post treatment.
The treatment effects on the appearance were also observed. Figure 2 shows the appearance of a WMS-affected marmoset. Before treatment, tabefaction, arched back, rough fur (Figure 2A), and alopecia (Figure 2B) were observed, whereas no abnormal appearance was seen after treatment (Figure 2C,D).
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/66768/66768fig01.jpg" /> 
Figure 1: Changes (pre- vs post-treatment) in hematocrit, serum albumin, and body weight in a marmoset receiving the modified treatment. (A) Hematocrit values, (B) serum albumin values, and (C) body weight. The dashed lines indicate the criteria values. Abbreviations: Htc = hematocrit; Alb = serum albumin; BW = body weight. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66768/66768fig01large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/66768/66768fig02.jpg" /> 
Figure 2: Representative appearance of WMS-affected marmoset. (A) Appearance of pretreatment animal: tabefaction, arched back, and rough fur are observed. (B) Tail of pretreatment animal: alopecia is observed. (C) Appearance of posttreatment animal. (D) Tail of posttreatment animal. Abbreviation: WMS = wasting marmoset syndrome. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66768/66768fig02large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>

Author Spotlight: Developing a Safer and More Efficient Treatment Protocol for Wasting Marmoset Syndrome (WMS)

Here, we describe the modified treatment methods for wasting marmoset syndrome (WMS, also known as inflammatory bowel disease (IBD)-like disease) with tranexamic acid. We also present how to administer the therapeutic agents orally, subcutaneously, and intravenously.

Modification of the Treatment Methods for Wasting Marmoset Syndrome with Tranexamic Acid and Supportive Measures

Wasting marmoset syndrome (WMS), a serious disease in captive common marmoset (Callithrix jacchus) colonies, is associated with a high mortality rate. The ...

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775 Views.  RIKEN Center for Brain Science. Here, we describe the modified treatment methods for wasting marmoset syndrome (WMS, also known as inflammatory bowel disease (IBD)-like disease) with tranexamic acid. We also present how to administer the therapeutic agents orally, subcutaneously, and intravenously.

Video: Author Spotlight: Developing a Safer and More Efficient Treatment Protocol for Wasting Marmoset Syndrome (WMS)

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 ...

Network Analysis of Foramen Ovale Electrode Recordings in Drug-resistant Temporal Lobe Epilepsy Patients

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control seizures. However, a significant minority of patients continues to exhibit post-operative seizures, even in those cases in which the suspected source of seizures has been correctly localized and resected. The protocol presented here combines a clinical procedure routinely employed during the pre-operative evaluation of temporal lobe epilepsy (TLE) patients with a novel technique for network analysis. The method allows for the evaluation of the temporal evolution of mesial network parameters. The bilateral insertion of foramen ovale electrodes (FOE) into the ambient cistern simultaneously records electrocortical activity at several mesial areas in the temporal lobe. Furthermore, network methodology applied to the recorded time series tracks the temporal evolution of the mesial networks both interictally and during the seizures. In this way, the presented protocol offers a unique way to visualize and quantify measures that considers the relationships between several mesial areas instead of a single area.

This protocol describes a procedure to track the evolution of mesial network measures in temporal lobe epilepsy (TLE) patients. It is based on the combination of intracranial recordings with a novel numerical technique for data analysis. Specifically, we present a protocol for network analyses of foramen ovale recordings.

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control ...

Oleic Acid-Injection in Pigs As a Model for Acute Respiratory Distress Syndrome

The acute respiratory distress syndrome is a relevant intensive care disease with an incidence ranging between 2.2% and 19% of intensive care unit patients. Despite treatment advances over the last decades, ARDS patients still suffer mortality rates between 35 and 40%. There is still a need for further research to improve the outcome of patients suffering from ARDS. One problem is that no single animal model can mimic the complex pathomechanism of the acute respiratory distress syndrome, but several models exist to study different parts of it. Oleic acid injection (OAI)-induced lung injury is a well-established model for studying ventilation strategies, lung mechanics and ventilation/perfusion distribution in animals. OAI leads to severely impaired gas exchange, deterioration of lung mechanics and disruption of the alveolo-capillary barrier. The disadvantage of this model is the controversial mechanistic relevance of this model and the necessity for central venous access, which is challenging especially in smaller animal models. In summary, OAI-induced lung injury leads to reproducible results in small and large animals and hence represents a well-suited model for studying ARDS. Nevertheless, further research is necessary to find a model that mimics all parts of ARDS and lacks the problems associated with the different models existing today.

In this article, we present a protocol to induce acute lung injury in pigs by central-venous injection of oleic acid. This is an established animal model for studying the acute respiratory distress syndrome (ARDS).

The acute respiratory distress syndrome is a relevant intensive care disease with an incidence ranging between 2.2% and 19% of intensive care unit ...

Transaxillary First Rib Resection for Treatment of the Thoracic Outlet Syndrome

Thoracic outlet syndrome (TOS) is a common disorder that causes a significant loss of productivity. The transaxillary first rib resection (TFRR) protocol has been used for the decompression of trapped neurovascular structures in the TOS. Among the other surgical procedures, the advantage of the TFRR is that it has the smallest rate of recurrence and better cosmetic outcomes. The disadvantage of TFRR is that it provides a narrow, and deep working corridor that makes obtaining vascular control challenging.

Here, we present a protocol of the transaxillary resection of the first rib for treatment of thoracic outlet syndrome caused by compression of the brachial plexus, subclavian vein and artery.

Thoracic outlet syndrome (TOS) is a common disorder that causes a significant loss of productivity. The transaxillary first rib resection (TFRR) protocol ...

Surfactant Depletion Combined with Injurious Ventilation Results in a Reproducible Model of the Acute Respiratory Distress Syndrome (ARDS)

Various animal models exist to study the complex pathomechanisms of the acute respiratory distress syndrome (ARDS). These models include pulmo-arterial infusion of oleic acid, infusion of endotoxins or bacteria, cecal ligation and puncture, various pneumonia models, lung ischemia/reperfusion models and, of course, surfactant depletion models, among others. Surfactant depletion produces a rapid, reproducible deterioration of pulmonary gas exchange and hemodynamics and can be induced in anesthetized pigs using repeated lung lavages with 0.9% saline (35 mL/kg body weight, 37 &#176;C). The surfactant depletion model supports investigations with standard respiratory and hemodynamic monitoring with clinically applied devices. But the model suffers from a relatively high recruitability and ventilation with high airway pressures can immediately reduce the severity of the injury by reopening atelectatic lung areas. Thus, this model is not suitable for investigations of ventilator regimes that use high airway pressures. A combination of surfactant depletion and injurious ventilation with high tidal volume/low positive end-expiratory pressure (high Tv/low PEEP) to cause ventilator induced lung injury (VILI) will reduce the recruitability of the resulting lung injury. The advantages of a timely induction and the possibility to perform experimental research in a setting comparable to an intensive care unit are preserved.

Surfactant Depletion Combined with Injurious Ventilation Results in a Reproducible Model of the Acute Respiratory Distress Syndrome ARDS

A combination of surfactant washout using 0.9% saline (35 mL/kg body weight, 37 &#176;C) and high tidal volume ventilation with low PEEP to cause moderate ventilator induced lung injury (VILI) results in experimental acute respiratory distress syndrome (ARDS). This method provides a model of lung injury with low/limited recruitability to study the effect of various ventilation strategies for extended periods.

Various animal models exist to study the complex pathomechanisms of the acute respiratory distress syndrome (ARDS). These models include pulmo-arterial ...

Laparoscopic Oocyte Retrieval and Cryopreservation during Vaginoplasty for Treatment of Mayer-Rokitansky-Kuster-Hauser Syndrome

In Mayer-Rokitansky-Kuster-Hauser Syndrome (MRKHS) patients who are scheduled for laparoscopic vaginoplasty and have a desire for biological motherhood, we propose that a concomitant laparoscopic oocyte retrieval for cryopreservation is performed. Oocyte retrieval is pursued at the beginning of the laparoscopy. Right and left 5 mm trocars are positioned, through which a 17 G ovum aspiration needle is used for puncture of the right and left ovaries, respectively. To facilitate exposure of the follicles, the ovaries are mobilized and held with laparoscopic forceps.
When aspirating multiple follicles near each other, the needle tip is retained in the ovary to reduce the number of times that the ovarian cortex is transfixed and due to the inherent risk of bleeding. Subsequent steps are unchanged compared to the Davydov laparoscopic modified technique for vaginoplasty. Prior to surgery, controlled ovarian stimulation is performed with a gonadotropin hormone-releasing hormone (Gn-RH) antagonist protocol, and the concomitant procedure of oocyte retrieval and vaginoplasty is scheduled 36 h after the final follicular maturation trigger. Follicular fluid is collected in the same 10 mL sterile tubes used during transvaginal oocyte retrieval and transferred in a warming block (37 &#176;C) to the assisted reproduction laboratory, where mature (metaphase II) oocytes are vitrified.
In this case, a series of 23 women with MRKH, oocytes were successfully retrieved and cryopreserved in all patients; vaginoplasty was subsequently conducted without modifications, and the inpatient and outpatient postoperative care (day of urinary catheter removal, day of hospital discharge, dilator use, and comfort at follow-up) remained unaffected. One postoperative complication occurred in one patient (fever developing on day 5 post surgery and intraperitoneal fluid detection on transabdominal ultrasound) and resolved after conservative treatment. Rather than performing surgical vaginoplasty and delaying oocyte retrieval in MRKH patients, this approach combines both procedures in a single laparoscopy, thereby minimizing surgical invasiveness and anesthesiologic risks.

A dedicated team can offer Mayer-Rokitansky-Kuster-Hauser patients the option to perform controlled ovarian stimulation and oocyte cryopreservation at the time of laparoscopic vaginoplasty.

In Mayer-Rokitansky-Kuster-Hauser Syndrome (MRKHS) patients who are scheduled for laparoscopic vaginoplasty and have a desire for biological motherhood, ...

Microscopic Cyst Resection for the Treatment of Patients Diagnosed with Epididymal Cyst

Epididymal cysts mostly occur in men aged 20-40 years old. Previous reports have covered concerns about postoperative complexes, including postoperative asoedema, hematoma, sustaining pain, and seminal tract obstruction in patients who have undertaken nonmicroscopic epididymal cyst resection or epididymal resection. Nonmicroscopic epididymal cyst surgery is suggested for patients with childbirth plans as a precaution. The treatment of male epididymal cysts via microtechnology is obviously a beneficial option; we took the lead in carrying out microscopic epididymal exploration and cyst resection surgery in China. From September 2017 to April 2021, 41 young and middle-aged male patients diagnosed with epididymal cysts underwent microtechnology treatment in a program titled &#34;microscopic epididymal exploration and cystectomy&#34;. The postoperative follow-up lasted for 3-50 months. The results confirmed that, as microscopic manipulation largely improved visualization of the subtle tissue structures of the epididymis, the cyst could be clearly dissected apart and completely removed intact under the microscope. Bleeding during the operation was significantly reduced (2-3 mL) and wound drainage was not required. According to follow-up data, microscopic treatment significantly reduced the incidence of postoperative scrotal hematoma, edema, and long-term postoperative pain, thereby promising a higher surgical success rate as well as recurrence prevention. Besides, preliminary experience and reflection suggest that microscopic epididymal exploration and cystectomy provide efficient preservation of the epididymal patency through refined treatment, while a better prognosis can be achieved. We recommend that surgery be carried out before the epididymal cyst develops to 0.8 cm in diameter, for fear that a larger epididymal cyst (&#62;0.9 cm in diameter) could cause the complete destruction of all tubules of the ipsilateral epididymis - a more severe case with damage to the testicular output network.

Here, we present a protocol to treat patients diagnosed with an epididymal cyst using microscopic cyst resection. Based on microscopic manipulation, a largely improved visualization of the subtle tissue structures of the epididymis was obtained, so the cyst could be clearly dissected apart and completely removed intact.

Epididymal cysts mostly occur in men aged 20-40 years old. Previous reports have covered concerns about postoperative complexes, including postoperative ...

Technique for Salivary Gland Ultrasound Examination of Sjogren's Syndrome

Ultrasonographic Evaluation of Salivary Glands for Sjogren's Syndrome: Diagnostic and Monitoring Insights

Sjogren's syndrome (SS) is a chronic autoimmune condition commonly affecting the exocrine glands, causing oral or ocular dryness and extraglandular manifestations including arthralgia, cytopenia, and lymphoma. The presence of autoantibodies against SSA/Ro, labial salivary gland biopsy, ocular staining, Schirmer's test, or salivary flow assessment are included in the current classification criteria of SS. However, the availability and invasiveness of these procedures limit their widespread use in clinical settings. Salivary gland ultrasonography (SGUS) is a non-invasive imaging modality for the evaluation of the salivary gland parenchyma and is increasingly utilized to aid diagnosis and monitoring in SS. 
This article presents the protocol of SGUS for image acquisition at the parotid and submandibular glands. The objective is to present a standardized, reproducible, and practical approach to diagnostic SGUS for SS in daily clinical settings. Major salivary glands are scanned in a stepwise approach, beginning at the angle of the mandible for the superficial lobe of the parotid gland, followed by the deep lobe below the ramus of the mandible. Submandibular areas are then scanned for the submandibular glands. The steps in obtaining salivary gland images at each anatomical site are explained in the accompanying video. The echogenicity and echotexture at the thyroid gland are taken as a reference. The homogeneity, the presence and distribution of hypoechoic areas within the glands, and the border of the salivary glands are examined. The sizes and features of intra-/peri-glandular lymph nodes are recorded. The most distinctive sonographic feature in SS is glandular heterogeneity with the presence of hypoechoic/hyperechoic areas within the glands. 
In summary, while SGUS cannot diagnose SS on its own, it can supplement the current classification criteria of SS and guide the clinical decision for salivary gland biopsy to support the diagnosis of SS in patients with sicca syndrome or suspicious systemic features, combined with autoantibody testing.

Author Spotlight: A Focus on Standardized Salivary Gland Ultrasound Protocol in Connective Tissue Disease Research

Salivary gland ultrasonography is a promising tool for the diagnosis and evaluation of disease activity and complications in Sjogren's syndrome (SS). The scanning technique for salivary gland ultrasound examination relevant to SS is described in the article.

Sjogren's syndrome (SS) is a chronic autoimmune condition commonly affecting the exocrine glands, causing oral or ocular dryness and extraglandular ...

Seminal Vesiculoscope Laser-Assisted Surgery to Treat Ejaculatory Duct Blockage

Treatment of Ejaculatory Duct Obstruction by Seminal Vesiculoscopy Assisted Flow Modification

Transurethral resection of ejaculatory duct (TURED) is a primary surgical approach to treat ejaculatory duct obstruction (EDO) caused by the ejaculatory duct cyst. Intraoperative excision of the verumontanum is usually required to expose the ejaculatory ducts. However, preserving the verumontanum structure allows for a better simulation of normal physiological anatomy. Maintaining the verumontanum may increase the risk of postoperative distal ejaculatory duct scarring, leading to recurrent obstruction or reduced semen volume. Therefore, we attempted a novel technique that preserves the verumontanum, which is relatively easier and safer compared to TURED. The following were the procedural steps: 1. A 6F seminal vesiculoscope was introduced through the external urethral orifice to the vicinity of the verumontanum, locating the opening of the affected-side ejaculatory duct and introducing a guidewire into the cyst. This successful step preserved the verumontanum, maximizing the retention of the anti-reflux mechanism in the distal ejaculatory duct. 2. The holmium laser enlarged the affected-side ejaculatory duct opening to 5 mm, decreasing the likelihood of postoperative closure of the ejaculatory duct opening and simplifying the procedure. 3. A window was created within the cyst to access the contralateral seminal vesicle, and then a holmium laser was used to burn and dilate the opening to 5 mm, redirecting the contralateral ejaculatory duct into the cystic cavity. This modification preserved the opening of the healthy-side ejaculatory duct and provided a new outflow passage for semen, reducing the risk of decreased semen volume postoperatively. The patients experienced no complications postoperatively, had shorter hospital stays, and showed improvement in semen volume. Hence, this surgical approach is simple yet effective.

Author Spotlight: Advanced Treatment of Seminal Duct Blockage Employing Endoscopy-Mediated Semen Channel Refluviation 

This protocol introduces the clinical application of seminal vesicle endoscopy combined with holmium laser in the treatment of ejaculatory duct obstruction caused by ejaculatory duct cyst.

Transurethral resection of ejaculatory duct (TURED) is a primary surgical approach to treat ejaculatory duct obstruction (EDO) caused by the ejaculatory ...

Enhancing Melasma Treatment with Tranexamic Acid and Microneedling Therapy

Roller Microneedle Combined with Tranexamic Acid Solution in Treating Melasma

Melasma, a common, acquired facial pigmentation skin disorder, presents a straightforward clinical diagnosis but poses challenges in terms of effective management. The precise underlying causes of melasma remain elusive, and the current therapeutic approaches predominantly encompass pharmaceutical and laser interventions, with limited efficacy. Transdermal administration stands as a prevalent treatment method for melasma, often facilitated by the application of microneedles. Among these, tranexamic acid emerges as a frequently employed therapeutic agent. A subset of microneedles, known as roller microneedles, plays a significant role in this approach by delicately puncturing the epidermis with multiple fine needles, synergizing with drug delivery. This methodology not only enhances drug absorption but also augments treatment efficacy while minimizing tissue trauma. These attributes forecast promising avenues for the treatment of melasma. This article primarily introduces the combination of roller microneedle and tranexamic acid solution in the treatment of melasma and demonstrates the efficacy of roller microneedle and tranexamic acid solution in the treatment of melasma through clinical cases.

Author Spotlight: Non-Surgical Treatment of Melasma&#8211; Microneedling with Tranexamic Acid

This article presents a protocol to conduct the treatment of melasma by using roller microneedles in combination with tranexamic acid solution and evaluate the efficacy.

Melasma, a common, acquired facial pigmentation skin disorder, presents a straightforward clinical diagnosis but poses challenges in terms of effective ...