The authors would like to thank professors Piccoli and Pastori of the Department of Medical and Surgical Sciences, University of Padova, Italy, for providing the BIVA software. Also, to Dr. Luis Llorente and Dra. Andrea Hinojosa-Azaola from the Department of Immunology and Rheumatology at the INCMNSZ for rheumatological assessment of patients. This work was supported by the CONACyT which sponsored the scholarship CVU 777701 for Mariel Lozada Mellado during his Ph.D. course study and through the Research Project Grant 000000000261652. The sponsor did not have any role in the study design or in the collection, analysis, or interpretation of data, nor in the writing of the report and in the decision to submit the paper for publication.

The present protocol was approved by and followed the Human Research and Ethics Committee guidelines of the National Institute of Medical Sciences and Nutrition Salvador Zubir&#225;n (Ref.: 1347). Informed consent was obtained from the human participants before participation in this study. Only patients in functional class I to III without total or partial arthroplasties18,19 and who were not candidates for prostheses were included in this study. Exclusion criteria included patients with cardiovascular disease, cancer, chronic kidney disease, pregnancy, or other autoimmune diseases that overlap with RA.
1. Recruitment of participants
<ol>
	<li>Recruit patients. 
	NOTE: For the present study, twelve women with RA were recruited from the rheumatology outpatient clinic.</li>
	<li>Ensure that the patients receive stable pharmacological treatment during the previous 6 months; which could include any of the following: antimalarial drugs (e.g., chloroquine, hydroxychloroquine), disease-modifying antirheumatic drugs (DMARD) (e.g., methotrexate, leflunomide), and glucocorticoids (e. g., prednisone)20. 
	&#8203;NOTE: According to the assessment by the rheumatologist, changes in pharmacological treatment could be made during the intervention period, if necessary.</li>
</ol>
2. Participant pre-test
NOTE: Pre-tests were performed 1 week before beginning the dynamic exercise program. Multifrequency bioelectrical impedance analysis equipment (see Table of Materials) was used, and measurements were performed with patients fasting for 4 to 5 h.
<ol>
	<li>Steps before testing
	<ol>
		<li>Ensure that these measurements are carried out by a standardized person with extensive experience.</li>
		<li>Clean the equipment using 0.05% chlorhexidine and ensure that the hands are washed.</li>
		<li>Explain the procedure to the patient and obtain the measurements for the weight (kg) and height (cm).</li>
		<li>Ask the patient to remove both shoes and their right sock as well as any metal objects that are in contact with their skin.</li>
		<li>Place the patient in a supine position for 5 min with both legs and arms extended and verify that they are not in contact with any part of their body.</li>
	</ol>
	</li>
	<li>Measurement of BIA
	<ol>
		<li>Clean the back of the hand and the right foot with 70% alcohol.</li>
		<li>Place two electrodes on the back of the hand: one on the third metacarpophalangeal joint and the other in the middle of the wrist at the level of the head of the ulna.</li>
		<li>Place two electrodes on the right foot: one at the third metatarsophalangeal joint and the other between the medial and lateral malleoli. There should be a gap of 5-10 cm between the electrodes.</li>
		<li>Connect the four cables of the equipment. Once connected, place the red clamps on the electrodes close to the fingernail and toenail; place the black clamps on the remaining electrodes. The cables must not cross each other.</li>
		<li>The impedance values (Z) of four different frequencies (5, 50, 100, and 200 kHz) will be shown on the equipment screen. Note the resistance and reactance values for the 50 kHz frequency. These values will be necessary to classify patients with cachexia. 
		NOTE: Bioelectrical impedance analysis using tetrapolar multifrequency equipment provides accurate resistance and reactance values at a single frequency of 50 kHz as well as the ratio between the 200 kHz and 5 kHz impedance values (200/5 kHz).</li>
	</ol>
	</li>
	<li>Classification of cachexia by BIVA
	<ol>
		<li>Download the BIVA tolerance R-Xc graph software (see Table of Materials) and open it. 
		NOTE: The software is a spreadsheet that can be seen at the bottom of seven worksheets.</li>
		<li>Go to the second worksheet, Reference populations; choose a row that corresponds to the reference population; copy it; and paste it in row two, marked in yellow. 
		NOTE: The reference population is chosen according to the age range, race, sex, and BMI of the population to be evaluated.</li>
		<li>Go to the fifth worksheet, Subjects, and insert the patient&#39;s data in the second row: in column A, enter the patient&#39;s ID; in column B, enter the number one; and for the next two columns, one can choose whether to enter the patient&#39;s name.</li>
		<li>In column E, enter the patient&#39;s sex, using M for men or F for women. In columns F and G, insert the previously noted resistance and reactance values at 50 kHz. Enter the height (cm) and weight (kg) in the next two columns.</li>
		<li>In column J, enter the number corresponding to the reference population chosen in the second worksheet.</li>
		<li>Insert a number between 1 and 10 in column K. It will be needed for the &#34;Point graph&#34; sheet; in the next column, enter the patient&#39;s age. 
		NOTE: One can choose values between 1 and 10 because there can be up to 10 patients to graph simultaneously in the BIVA tolerance software.</li>
		<li>The options bar is at the top of the software. Find the complements option and click it. Then, select the calculate option that will be displayed and click it. Observe the resistance and reactance values adjusted by height and phase angle.</li>
		<li>Next, navigate to sheet 3, Point Graph, and observe a BIVA graph according to the chosen reference population. A dialogue box will be displayed. Select the group code entered in column K for step 2.3.6. Select OK, and the BIVA graph will be shown with the patient&#39;s vector drawn as a geometric figure.</li>
		<li>Observe the tolerance ellipses of 50%, 75%, and 95% as well as the quadrants I, II, III, and IV in the BIVA graph. To classify a patient with cachexia using BIVA, the vector must be in the lower right quadrant (quadrant IV) and outside the 75% tolerance ellipse (Figure 1). 
		NOTE: Patients whose vectors fall in any of the quadrants within the &#60;75% tolerance ellipses will be considered with a normal body composition classification21.</li>
	</ol>
	</li>
</ol>
3. Dynamic exercise program
NOTE: The program was applied and supervised by a physiotherapist. An intervention duration of 48 sessions per patient was estimated. The exercise sessions were carried out in a mechanotherapy gym within a physiotherapy area belonging to the Rheumatology and Immunology department of the &#34;INCMNSZ&#34; with a duration of 90 min, twice a week.
<ol>
	<li>Evaluation of the session
	<ol>
		<li>Ask patients about the pain or discomfort they perceive in their joints. 
		NOTE: The visual analogue scale (VAS) was used to assess pain. If they reported pain on the VAS above 7/10 in any joint, a more specific evaluation was carried out by the physiotherapy department (e.g., electrotherapy was used if there was only pain, thermotherapy was used if there was stiffness present, and cryotherapy was used when there was both pain and inflammation).</li>
		<li>Take vital signs before each exercise session.</li>
	</ol>
	</li>
	<li>Warm-up 
	NOTE: With a duration of 15 min, a general dynamic warm-up divided into phases was established. Activation phase: simple, gentle, and global movements were performed for all movement arcs while remaining in a static position, with 10 to 15 repetitions. Set-up phase: in this last part, gentle dynamic exercises were performed, which simulated the gestures of the movements that would be implemented in the work phase, with 10-15 repetitions.
	<ol>
		<li>Activation phase
		<ol>
			<li>Choose the most suitable warm-up exercise, including upper- and lower-extremity joints and their range of motion.
			<ol>
				<li>Upper extremity: Instruct the patient to reach a range of motion with no discomfort for each joint movement. The instructor must lead the patient through a normal-speed movement and instruct the patient to avoid a painful range of motion.</li>
				<li>Lower extremity: Instruct the patient to perform the warm-up in a standing position with both feet on the ground and on a stable surface. Instruct the patient to reach a non-painful movement speed through the range of motion for each joint while the patient sits in a chair. 
				NOTE: If some patients can stand for a long time, a sitting position must be reached, considering a stable chair with the back straight and the feet on the ground. The available ranges of motion of the hip, knee, ankle, and feet must be included.</li>
			</ol>
			</li>
		</ol>
		</li>
		<li>Set-up phase
		<ol>
			<li>Instruct the patient to perform functional movement patterns that include more than two joints per segment (lower limb or upper limb).</li>
			<li>Give supervision during this stage to bring a sense of wellness during the movement and adjust the range of motion when the patient presents discomfort.</li>
		</ol>
		</li>
	</ol>
	</li>
	<li>Work phase 
	NOTE: With a duration of 60 min, the work phase is divided into three stages of 20 min each.
	<ol>
		<li>Aerobic: perform the work on a treadmill. 
		NOTE: Select a treadmill with no default inclination.
		<ol>
			<li>Ensure the emergency stop device is working correctly and explain the safety measures to the patient. Advise the patient to wear sports shoes.</li>
			<li>Give the patients information about the adaptations that must be performed when the treadmill starts and must be performed properly to avoid unnatural gait movements.</li>
			<li>Establish a base speed for each patient, asking for a normal feeling during walking.</li>
			<li>Adjust the speed after 5 min on the treadmill. Using a pulse oximeter (see Table of Materials), measure the heart rate while the speed is increased until reaching a heart rate zone between 55% and 75%14,31of the HRmax. 
			NOTE: If the patient&#39;s heart rate goes over the 75% HRmax, the speed must be reduced to the ideal heart rate zone. Instruct the patient to look for a comfortable pace.</li>
			<li>After 10 min, ask the patient for an assessment using a perceived effort rating scale. 
			NOTE: The modified Borg rating of the perceived exertion scale was used to assess perceived exertion.</li>
			<li>Lower the treadmill speed to a comfortable pace for the patient&#39;s last 5 min. The speed must be lowered to a total stop when reaching 5 min.</li>
			<li>Ask the patient for any pain or discomfort after the use of the treadmill.</li>
		</ol>
		</li>
		<li>Resistance exercises 
		NOTE: Directed joint mobility exercises were used in combination with muscle strength exercises. The workout consisted of a set of 8-10 repetitions per exercise. Soft (0.5-2.6 kg) and medium (0.7-3.2 kg) resistance bands were used, and the resistance was gradually increased every 2 weeks. The dosage of the exercise depended on the state of the patient at the time of the intervention.
		<ol>
			<li>Upper Extremity
			<ol>
				<li>Instruct the patient to perform upper-extremity mobility while handling a wooden stick (&#60;1 kg) with both hands.</li>
				<li>Teach the patient combined exercises that include the range of motion of more than two joints (e.g., shoulder and elbow flexion).</li>
				<li>Instruct the patient to hold a band above the ends. The patient must roll up their hand with the end of the band to ensure their grip. 
				NOTE: If the patient&#39;s hands have discomfort, the instructor must gently secure the band to their wrist.</li>
				<li>Instruct the patient to put one end of the band on the floor and step it with their foot. Then, perform elbow flexion against the band&#39;s resistance. Elbow extension must work on eccentric contraction while returning to the neutral position. 
				NOTE: The patient must be standing with a stable foot base and good posture. If the patient indicates some discomfort, this exercise must be performed in a sitting position.</li>
				<li>Instruct the patient to roll up a band on their hand, ensuring no excessive pressure is applied. The other end should be held by the free hand of the patient next to the body at hip level. Then, instruct the patient to flex the elbow at 90&#176; with the elbow in a neutral position. 
				NOTE: The patient can rest for 20 s between movements.</li>
			</ol>
			</li>
			<li>Lower extremity
			<ol>
				<li>Instruct the patient to sit in a stable chair with 90&#176; hip and knee flexion and tie the ends of the resistance band, making a loop band. The patient must surround their legs with the rubber band at the distal part of the femur (above the knee). In this position, instruct the patient to perform hip flexions for each leg up to 20 to 30 degrees above the starting position. 
				NOTE: For correct alignment, avoid hip rotation and knee flexion. If the patient indicates discomfort, reduce the range of motion.</li>
				<li>Instruct the patient to sit in a stable chair with 90&#176; hip and knee flexion and tie the ends of the resistance band, making a loop band. The patient must surround their legs with the rubber band at the distal part of the femur (above the knee). In this position, instruct the patient to perform a slight hip flexion (above 10&#176; from the base position) and hip abduction. 
				NOTE: For correct alignment, avoid hip rotation and excessive knee flexion. If the patient indicates discomfort, reduce the range of motion.</li>
				<li>Instruct the patient to sit in a stable chair with 90&#176; hip and knee flexion and tie the ends of the resistance band, making a loop band. The patient must surround the nearest chair leg and their own leg with the rubber band at the ankle. Instruct the patient to return, at a slow tempo, to the base position. 
				NOTE: For correct alignment, the patient must maintain a comfortable sitting position and avoid hip flexion compensation. If needed, the patient can hold the base of the chair with their hands for more stability. The steps can be performed with one leg at a time or by changing sides.</li>
				<li>Instruct the patient to hold a standing position. Then, ask the patient to tie the ends of the resistance band, make a loop band, and place the band around their ankles. Instruct the patient to perform reps of changing positions between sitting and standing. 
				NOTE: If the patient feels discomfort during the exercise, reassess and make the exercise easy by using a higher chair to reduce knee flexion or using a second chair where the patient can support himself and facilitate movement.</li>
			</ol>
			</li>
		</ol>
		</li>
		<li>Recreation games 
		NOTE: Consisting of the implementation of exercise series that involve gestures or movements adapted from a particular sport such as soccer, basketball, or volleyball, integrating flexibility and coordination components, 4 to 7 stations consisting of polyarticular movements and different exercises are created, and two series of 8 to 15 repetitions are worked (with difficulty increasing every 2 weeks).
		<ol>
			<li>Choose the most suitable exercise based on a sporting gesture for the patients each session and make an exercise station. Each station must be designed taking into account the limitations of the patient.</li>
			<li>Make a soccer goal with two chairs with a 1.3 m separation between them.</li>
			<li>Instruct the patients to hit a 30 cm plastic ball with their feet at a 3 m spot in front of the soccer goal.</li>
			<li>Control the difficulty by increasing the repetitions or sets per station and by adding new stations to the circuit. 
			NOTE: Example station designs: (1) Attach a &#34;Ula Ula&#34; ring to the tip of a 1.3 m wooden stick, place the patient at a 2 m throwing point in front of the ring, and instruct them to throw a 30 cm plastic ball with their arms to the &#34;Ula Ula ring&#34;. Each patient must score at least 5 times and can score up to 10 times. (2) Attach a rope along the room walls to simulate a volleyball net. The rope must have a minimum height of 1. 7 m, and two patients must be in position on each side. Instruct the patients to pass a 40 cm air balloon over the rope at least 10 to 15 times each. (3) Place two patients with a 3 m separation between them and instruct the patients to throw a 30 cm plastic ball with their arms. Each patient must throw the plastic ball at least 10 times per arm. Patients must always supervise.</li>
		</ol>
		</li>
	</ol>
	</li>
	<li>Cooling 
	&#8203;NOTE: Cooling has a duration of 15 min and is composed of active static stretches.
	<ol>
		<li>Applied globally, stretching must be performed gently without putting stress on the joint. Stretching should not produce discomfort for the patient.</li>
		<li>Maintain each stretch for 15 to 20 s.</li>
	</ol>
	</li>
</ol>
4. Post-test evaluation
NOTE: The post-test assessment must be scheduled during the week after the last exercise session.
<ol>
	<li>Repeat the measurement of body composition to obtain the BIVA classification, as described in the pre-test. 
	NOTE: To make a comparison between before and after the implementation of the dynamic exercise program, it is necessary to obtain the mean of the resistance difference divided by the height (dR/H), the mean of the reactance difference divided by the height (dXc/H), and the standard deviation and the Pearson correlation coefficient of the differences with the following equation8: <img alt="Equation 1" src="/files/ftp_upload/65692/65692eq01.jpg" style="margin: auto;" /></li>
	<li>To obtain the change in the resistance and reactance, download the BIVA confidence software (see Table of Materials) and open it. 
	NOTE: The software is a spreadsheet; at the bottom, you can see five worksheets.</li>
	<li>In the fourth worksheet, &#34;Paired data&#34;, check for ten columns where it will be necessary to insert the requested data.
	<ol>
		<li>In column A, place the group ID. In column B, place the number of patients that were evaluated.</li>
		<li>In column C, insert the mean of d R/H obtained previously. In the next column, add the standard deviation.</li>
		<li>In column E, insert the mean of d Xc/H, and in the following column insert the standard deviation. In column G, insert the correlation coefficient obtained previously. 
		NOTE: In column H, choose to place 1, where one can display the confidence ellipse on the chart, or option 2 if you want to show the confidence ellipse and the difference mean vector.</li>
		<li>In the following two columns, one can choose whether to place the names of the group and the equipment that was used to make the measurements.</li>
	</ol>
	</li>
	<li>Once all the necessary data are complete, go to sheet 5, &#34;Paired graph&#34;. There, a graph of the means of the difference is visible, and will be able to locate the vector of the resistance and reactance mean, in addition to the confidence ellipse.</li>
	<li>To evaluate whether the change is statistically significant, locate the complements option in the toolbar and click it. It will open a box with Hotelling&#39;s T2 test statistic8, allowing one to locate the value of p.</li>
</ol>

The video discusses research on using vector impedance analysis to assess hydration changes in body cell mass in rheumatoid arthritis patients. It highlights the increased infection risk associated with low body cell mass and emphasizes the benefits of physical exercise in improving strength, quality of life, and inflammation in these patients.

Developing an Exercise Program for Rheumatoid Arthritis Patients

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J., Carmona, L., Hern&#225;ndez-Garc&#237;a, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estudio+sobre+el+manejo+de+la+artritis+reumatoide+en+Espa&#241;a+(emAR+II)+[Study+on+the+management+of+rheumatoid+arthritis+in+Spain+(emAR+II)].+Caracter&#237;sticas+cl&#237;nicas+de+los+pacientes+[Clinical+characteristic+of+patients].">Estudio sobre el manejo de la artritis reumatoide en Espa&#241;a (emAR II) [Study on the management of rheumatoid arthritis in Spain (emAR II)]. Caracter&#237;sticas cl&#237;nicas de los pacientes [Clinical characteristic of patients].</a> Reumatolog&#237;a Clinica. 8 (5), 236-242 (2012).</li><li>Hurkmans, E., Van der Giesen, F. J., Vlieland, T. P. M. V., Schoones, J., Van den Ende, E. C. H. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+exercise+programs+(aerobic+capacity+and/or+muscle+strength+training)+in+patients+with+rheumatoid+arthritis.">Dynamic exercise programs (aerobic capacity and/or muscle strength training) in patients with rheumatoid arthritis.</a> Cochrane Database of Systematic Reviews. 4, CD006853 (2009).</li><li>Baillet, 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=Efficacy+of+cardiorespiratory+aerobic+exercise+in+rheumatoid+arthritis:+Meta-analysis+of+randomized+controlled+trials.">Efficacy of cardiorespiratory aerobic exercise in rheumatoid arthritis: Meta-analysis of randomized controlled trials.</a> Arthritis Care &amp; Research. 62 (7), 984-992 (2010).</li><li>De Jong, Z., 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=Long-term+follow-up+of+a+high-intensity+exercise+program+in+patients+with+rheumatoid+arthritis.">Long-term follow-up of a high-intensity exercise program in patients with rheumatoid arthritis.</a> Clinical Rheumatology. 28 (6), 663-671 (2009).</li><li>Garc&#237;a-Morales, J. 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=Effect+of+a+dynamic+exercise+program+in+combination+with+Mediterranean+diet+on+quality+of+life+in+women+with+rheumatoid+arthritis.">Effect of a dynamic exercise program in combination with Mediterranean diet on quality of life in women with rheumatoid arthritis.</a> Journal of Clinical Rheumatology. 26 (2), S116-S122 (2019).</li><li>Munneke, 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=Effect+of+a+high-intensity+weight-bearing+exercise+program+on+radiologic+damage+progression+of+the+large+joints+in+subgroups+of+patients+with+rheumatoid+arthritis.">Effect of a high-intensity weight-bearing exercise program on radiologic damage progression of the large joints in subgroups of patients with rheumatoid arthritis.</a> Arthritis &amp; Rheumatism. 53, 410-417 (2005).</li><li>Hochberg, M., Chang, R., Dwosh, I., Lyndsey, S., Pincus, T., 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+American+College+of+Rheumatology+1991+Revised+Criteria+for+the+Classification+of+Global+Functional+Status+in+Rheumatoid+Arthritis.">The American College of Rheumatology 1991 Revised Criteria for the Classification of Global Functional Status in Rheumatoid Arthritis.</a> Arthritis &amp; Rheumatism. 35, 498-502 (1991).</li><li>Nikiphorou, E., Konan, S., MacGregor, A. J., Haddad, F. S., Young, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+surgical+treatment+of+rheumatoid+arthritis.">The surgical treatment of rheumatoid arthritis.</a> Bone Joint Journal. 96 (10), 1287-1289 (2014).</li><li>Jacqueline, B., 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=Rheumatoid+Arthritis:+A+Brief+Overview+of+the+Treatment.">Rheumatoid Arthritis: A Brief Overview of the Treatment.</a> Medical Principles and Practice. 27 (6), 501-507 (2019).</li><li>Piccoli, A., Rossi, B., Pillon, L., Bucciante, G. <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+method+for+monitoring+body+fluid+variation+by+bioimpedance+analysis:+the+RXc+graph.">A new method for monitoring body fluid variation by bioimpedance analysis: the RXc graph.</a> Kidney International. 46 (2), 534-539 (1994).</li><li>Benatti, F. B., Pedersen, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exercise+as+an+anti-inflammatory+therapy+for+rheumatic+diseases+-+Myokine+regulation.">Exercise as an anti-inflammatory therapy for rheumatic diseases - Myokine regulation.</a> Nature Reviews Rheumatology. 11 (2), 86-97 (2015).</li><li>Cooney, J. 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=Benefits+of+Exercise+in+Rheumatoid+Arthritis.">Benefits of Exercise in Rheumatoid Arthritis.</a> Journal of Aging Research. 6, 297-310 (2011).</li><li>Barbosa-Silva, M. C. G., Barros, 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=Bioelectrical+impedance+analysis+in+clinical+practice:+a+new+perspective+on+its+use+beyond+body+composition+equations.+Current+Opinion.">Bioelectrical impedance analysis in clinical practice: a new perspective on its use beyond body composition equations. Current Opinion.</a> Clinical Nutrition and Metabolic Care. 8 (3), 311-317 (2005).</li><li>Mulasi, U., Kuchnia, A. J., Cole, A. J., Earthman, C. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioimpedance+at+the+bedside:+current+applications,limitations,+and+opportunities.">Bioimpedance at the bedside: current applications,limitations, and opportunities.</a> Nutrition in Clinical Practice. 30 (2), 180-193 (2015).</li><li>Steihaug, O. M., Bogen, B., Kristoffersen, M., Ranhoff, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bones,+blood+and+steel:+How+bioelectrical+impedance+analysis+is+affected+by+a+hip+fracture+and+surgical+implants.">Bones, blood and steel: How bioelectrical impedance analysis is affected by a hip fracture and surgical implants.</a> Journal of Electrical Bioimpedance. 8, 54-59 (2017).</li><li>Nwosu, A. C., 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=Bioelectrical+impedance+vector+analysis+(BIVA)+is+a+method+to+compare+body+composition+differences+according+to+cancer+stage+and+type.">Bioelectrical impedance vector analysis (BIVA) is a method to compare body composition differences according to cancer stage and type.</a> Clinical nutrition ESPEN. 30, 59-66 (2019).</li><li>Martins, P. C., Gobbo, L. A., Silva, D. A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioelectrical+impedance+vector+analysis+(BIVA)+in+university+athletes.">Bioelectrical impedance vector analysis (BIVA) in university athletes.</a> Journal of the International Society of Sports Nutrition. 18 (7), 1-8 (2021).</li><li>Norman, K., Pirlich, M., Sorensen, J., Christensen, P., Kemps, M., Sch&#252;tz, T., Lochs, H., Kondrup, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioimpedance+vector+analysis+as+a+measure+of+muscle+function.">Bioimpedance vector analysis as a measure of muscle function.</a> Clinical Nutrition. 28 (1), 78-82 (2009).</li><li>Stagi, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Usability+of+classic+and+specific+bioelectrical+impedance+vector+analysis+in+measuring+body+composition+of+children.">Usability of classic and specific bioelectrical impedance vector analysis in measuring body composition of children.</a> Clinical nutrition. 41 (3), 673-679 (2022).</li><li>Garber, C. E., 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=American+College+of+Sports+Medicine.+American+College+of+Sports+Medicine+position+stand.+Quantity+and+quality+of+exercise+for+developing+and+maintaining+cardiorespiratory,+musculoskeletal,+and+neuromotor+fitness+in+apparently+healthy+adults:+guidance+for+prescribing+exercise.">American College of Sports Medicine. American College of Sports Medicine position stand. Quantity and quality of exercise for developing and maintaining cardiorespiratory, musculoskeletal, and neuromotor fitness in apparently healthy adults: guidance for prescribing exercise.</a> Medicine &amp; Science in Sports &amp; Exercise. 43 (7), 1334-1359 (2011).</li></ol>

The authors have nothing to disclose.

In rheumatoid arthritis, the vicious circle of the disease has been described, which refers to the structural changes in joints caused by inflammation mechanisms; these changes, together with the chronic inflammatory state, lead to patients going through stages of great pain and inflammation, with structural changes in joints and as a consequence functional disability, that increase the risk of developing metabolic and cardiovascular diseases and alterations in body composition such as rheumatoid cachexia<sup class='xref...

Rheumatoid arthritis (RA) is a disabling disease that affects patients&#39; functionality and independence due to acute joint pain, reduced muscle strength, and impaired physical function, all of which are associated with the inflammatory process inherent to the disease1,2. In advanced stages, persistent inflammation causes structural alterations leading to deformity, joint dysfunction, and rheumatoid cachexia, which is a poor prognostic factor for these patients3,4.
Rheumatoid cachexia is characterized by alterations in body composition, such as muscle loss with stable weight and increased fat mass, which can significantly impact the quality of life for these patients3,5,6. Various techniques are available to assess body composition, with the most widely used being bioelectrical impedance analysis (BIA). However, when conventional BIA analysis is used in subjects with altered body compositions, the estimations may be limited as they are based on prediction equations formulated for a healthy or normally hydrated population7,8.
A different approach, called bioelectrical impedance vector analysis (BIVA), utilizes the impedance vector based on graphical RXc. It utilizes impedance, resistance (R), and reactance (Xc) data corrected for height, resulting in a vector that provides information about the hydration state and integrity of the cell mass. BIVA is capable of classifying patients into categories such as dehydration, overhydration, normal, athlete, lean, cachectic, and obese, making it a valuable tool for RA patients8,9,10. Vectors located above or below the main axis (the left or right halves of the ellipses) have been associated with higher and lower cell mass&#160;in soft tissues, respectively. Forward and backward displacements of vectors parallel to the major axis are linked to dehydration and fluid overload. Athletes are defined as individuals with higher cell mass, potentially accompanied by dehydration. The lean classification refers to those with lower cell mass, potentially accompanied by dehydration, and the obese classification applies to individuals with higher cell mass, which may be accompanied by fluid overload. The classification of cachexia by BIVA is determined by high resistance and low reactance values, represented by the movement of the vector to the right of the graph, indicating a decrease in cell mass, potentially accompanied by an alteration in hydration status11 (Figure 1).
Conventional pharmacological treatments for RA primarily focus on reducing pain, inflammation, and joint damage progression, with limited attention given to alterations in body composition12. Among the non-pharmacological therapies commonly used in this population, exercise-based interventions have shown positive outcomes in improving functionality, fatigue, pain, joint mobility, aerobic capacity, muscle strength, endurance, flexibility, and psychological well-being. Importantly, these interventions have been shown to achieve these benefits without exacerbating symptoms or causing joint damage in patients without extensive pre-existing damage13,14,15,16,17. However, there is limited knowledge on implementing and evaluating changes in hydration and body cell mass status following exercise interventions in this population. These patients often experience pain, inflammation, and structural joint changes, limiting the types of activities they can engage in and further complicating body composition assessments using traditional approaches. This protocol aims to demonstrate how to evaluate changes in hydration and body cell mass status using bioelectrical impedance vectorial analysis after implementing a dynamic exercise program for patients with rheumatoid arthritis. Additionally, the protocol provides details of the dynamic exercise program, including cardiovascular capacity, strength, and coordination components, as well as the steps, instruments, limitations, and general considerations.

<table><tbody><tr><td>Alcohol 70% swabs</td><td>NA</td><td>NA</td><td>Any brand can be used</td></tr><tr><td>bicycle ergometer</td><td>NA</td><td>NA</td><td>Any brand can be used</td></tr><tr><td>BIVA&amp;nbsp; tolerance software 2002</td><td>NA</td><td>NA</td><td>Is a sofware created for academic use, can be download in http://www.renalgate.it/formule_calcolatori/bioimpedenza.htm in &quot;LE FORMULE DEL Prof. Piccoli&quot; section</td></tr><tr><td>BIVA confidence software</td><td>NA</td><td>NA</td><td>Is a sofware created for academic use, can be download in http://www.renalgate.it/formule_calcolatori/bioimpedenza.htm in &quot;LE FORMULE DEL Prof. Piccoli&quot; section</td></tr><tr><td>Chair</td><td>NA</td><td>NA</td><td>Any brand can be used</td></tr><tr><td>Chlorhexidine</td><td>NA</td><td>NA</td><td>Any brand can be used, 0.05%</td></tr><tr><td>Examination table</td><td>NA</td><td>NA</td><td>Any brand can be used</td></tr><tr><td>Leadwires square socket</td><td>BodyStat</td><td>SQ-WIRES</td><td></td></tr><tr><td>Long Bodystat 0525 electrodes</td><td>BodyStat</td><td>BS-EL4000</td><td></td></tr><tr><td>Plastic ball</td><td>NA</td><td>NA</td><td>Any brand can be used, 30 cm</td></tr><tr><td>Pulse oximeter</td><td>NA</td><td>NA</td><td>Any brand can be used</td></tr><tr><td>Quadscan 4000&amp;nbsp; equipment</td><td>BodyStat</td><td>BS-4000</td><td>Impedance measuring range: 20 - 1300 &amp;Omega; ohms&lt;br/&gt; Test Current: 620 &amp;mu;A&lt;br/&gt; Frequency: 5, 50, 100, 200 kHz&lt;br/&gt; Accuracy: Impedance 5 kHz: +/- 2 &amp;Omega;&lt;br/&gt; Impedance 50 kHz: +/- 2 &amp;Omega;&lt;br/&gt; Impedance 100 kHz: +/- 3 &amp;Omega;&lt;br/&gt; Impedance 200 kHz: +/- 3 &amp;Omega;&lt;br/&gt; Resistance 50 kHz: +/- 2 &amp;Omega;&lt;br/&gt; Reactance 50 kHz: +/- 1 &amp;Omega;&lt;br/&gt; Phase Angle 50 kHz: +/- 0.2&amp;deg;&lt;br/&gt; Calibration: A resistor is supplied for independent verification from time to time. The impedance value should read between 496 and 503 &amp;Omega;.</td></tr><tr><td>Resistence bands</td><td>NA</td><td>NA</td><td>Any brand can be used, with resistence of 0.5 kg to 3.2 kg</td></tr><tr><td>Stationary bicycle</td><td>NA</td><td>NA</td><td>Any brand can be used</td></tr><tr><td>Treadmill</td><td>NA</td><td>NA</td><td>Any brand can be used</td></tr><tr><td>Wooden stick</td><td>NA</td><td>NA</td><td>Any brand can be used, 1.5m in large and &amp;lt;1kg</td></tr></tbody></table>

evaluation of changes in hydration and body cell mass with bioelectrical impedance analysis after exercise program for rheumatoid arthritis patients

Rheumatoid arthritis (RA) is a debilitating disease that can result in complications such as rheumatoid cachexia. While physical exercise has shown benefits for RA patients, its impact on hydration and body cell mass remains uncertain. The presence of pain, inflammation, and joint changes often restrict activity and make traditional body composition assessments unreliable due to altered hydration levels. Bioelectrical impedance is a commonly used method for estimating body composition, but it has limitations since it was primarily developed for the general population and does not consider changes in body composition. On the other hand, bioelectrical impedance vectorial analysis (BIVA) offers a more comprehensive approach. BIVA involves graphically interpreting resistance (R) and reactance (Xc), adjusted for height, to provide valuable information about hydration status and the integrity of the cell mass.
Twelve women with RA were included in this study. At the beginning of the study, hydration and body cell mass measurements were obtained using the BIVA method. Subsequently, the patients participated in a six-month dynamic exercise program encompassing cardiovascular capacity, strength, and coordination training. To evaluate changes in hydration and body cell mass, the differences in the R and Xc parameters, adjusted for height, were compared using BIVA confidence software. The results showed notable changes: resistance decreased after the exercise program, while reactance increased. BIVA, as a classification method, can effectively categorize patients into dehydration, overhydration, normal, athlete, thin, cachectic, and obese categories. This makes it a valuable tool for assessing RA patients, as it provides information independent of body weight or prediction equations. Overall, the implementation of BIVA in this study shed light on the effects of the exercise program on hydration and body cell mass in RA patients. Its advantages lie in its ability to provide comprehensive information and overcome the limitations of traditional body composition assessment methods.

Results are presented for six female patients with RA who participated in a 48 session dynamic exercise program. The mean age of the patients was 52.7 &#177; 13.1 years, and their BMI was 26.8 &#177; 4.6. The average disease duration was 15.5 &#177; 6.1 years, and the disease activity, measured by Disease Activity Score 28, was classified as low activity with an average of 1.9 &#177; 1. Regarding disability, the Health Assessment Questionnaire Disability yielded an average score of 0.5 &#177; 0.3. For the six participant...

Watch this Scientific Journal Video about Implementation of BIVA for Analyzing Disease Risk Factors in Patients with Low Body Cell Mass at JoVE.com

Author Spotlight: Implementation of BIVA for Analyzing Disease Risk Factors in Patients with Low Body Cell Mass 

This protocol assesses alterations in hydration and body cell mass status using bioelectrical impedance vectorial analysis following a dynamic exercise program designed for patients with rheumatoid arthritis. The dynamic exercise program itself is detailed, highlighting its components focused on cardiovascular capacity, strength, and coordination. The protocol details steps, instruments, and limitations.

Evaluation of Changes in Hydration and Body Cell Mass with Bioelectrical Impedance Analysis after Exercise Program for Rheumatoid Arthritis Patients

Rheumatoid arthritis (RA) is a debilitating disease that can result in complications such as rheumatoid cachexia. While physical exercise has shown ...

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1.1K Views.  National Autonomous University of Mexico. This protocol assesses alterations in hydration and body cell mass status using bioelectrical impedance vectorial analysis following a dynamic exercise program designed for patients with rheumatoid arthritis. The dynamic exercise program itself is detailed, highlighting its components focused on cardiovascular capacity, strength, and coordination. The protocol details steps, instruments, and limitations.

Video: Author Spotlight: Implementation of BIVA for Analyzing Disease Risk Factors in Patients with Low Body Cell Mass 

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

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

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

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

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

Evaluation of Cancer Stem Cell Migration Using Compartmentalizing Microfluidic Devices and Live Cell Imaging

In the last 40 years, the United States invested over 200 billion dollars on cancer research, resulting in only a 5% decrease in death rate. A major obstacle for improving patient outcomes is the poor understanding of mechanisms underlying cellular migration associated with aggressive cancer cell invasion, metastasis and therapeutic resistance1. Glioblastoma Multiforme (GBM), the most prevalent primary malignant adult brain tumor2, exemplifies this difficulty. Despite standard surgery, radiation and chemotherapies, patient median survival is only fifteen months, due to aggressive GBM infiltration into adjacent brain and rapid cancer recurrence2. The interactions of aberrant cell migratory mechanisms and the tumor microenvironment likely differentiate cancer from normal cells3. Therefore, improving therapeutic approaches for GBM require a better understanding of cancer cell migration mechanisms. Recent work suggests that a small subpopulation of cells within GBM, the brain tumor stem cell (BTSC), may be responsible for therapeutic resistance and recurrence. Mechanisms underlying BTSC migratory capacity are only starting to be characterized1,4.
Due to a limitation in visual inspection and geometrical manipulation, conventional migration assays5 are restricted to quantifying overall cell populations. In contrast, microfluidic devices permit single cell analysis because of compatibility with modern microscopy and control over micro-environment6-9.
We present a method for detailed characterization of BTSC migration using compartmentalizing microfluidic devices. These PDMS-made devices cast the tissue culture environment into three connected compartments: seeding chamber, receiving chamber and bridging microchannels. We tailored the device such that both chambers hold sufficient media to support viable BTSC for 4-5 days without media exchange. Highly mobile BTSCs initially introduced into the seeding chamber are isolated after migration though bridging microchannels to the parallel receiving chamber. This migration simulates cancer cellular spread through the interstitial spaces of the brain. The phase live images of cell morphology during migration are recorded over several days. Highly migratory BTSC can therefore be isolated, recultured, and analyzed further. 
Compartmentalizing microfluidics can be a versatile platform to study the migratory behavior of BTSCs and other cancer stem cells. By combining gradient generators, fluid handling, micro-electrodes and other microfluidic modules, these devices can also be used for drug screening and disease diagnosis6. Isolation of an aggressive subpopulation of migratory cells will enable studies of underlying molecular mechanisms.

A compartmentalizing microfluidic device for investigating cancer stem cell migration is described. This novel platform creates a viable cellular microenvironment and enables microscopic visualization of live cell locomotion. Highly motile cancer cells are isolated to study molecular mechanisms of aggressive infiltration, potentially leading to more effective future therapies.

In the last 40 years, the United States invested over 200 billion dollars on cancer research, resulting in only a 5% decrease in death rate. A major ...

Assessing Teratogenic Changes in a Zebrafish Model of Fetal Alcohol Exposure

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, including mental retardation due to disordered and damaged brain development. Fetal alcohol spectrum disorder (FASD) is a term used to cover a continuum of birth defects that occur due to maternal alcohol consumption, and occurs in approximately 4% of children born in the United States. With 50% of child-bearing age women reporting consumption of alcohol, and half of all pregnancies being unplanned, unintentional exposure is a continuing issue2. In order to best understand the damage produced by ethanol, plus produce a model with which to test potential interventions, we developed a model of developmental ethanol exposure using the zebrafish embryo. Zebrafish are ideal for this kind of teratogen study3-8. Each pair lays hundreds of eggs, which can then be collected without harming the adult fish. The zebrafish embryo is transparent and can be readily imaged with any number of stains. Analysis of these embryos after exposure to ethanol at different doses and times of duration and application shows that the gross developmental defects produced by ethanol are consistent with the human birth defect. Described here are the basic techniques used to study and manipulate the zebrafish FAS model.

In order to understand the molecular mechanisms of the ethanol-induced developmental damage, we have developed a zebrafish model of ethanol exposure and are exploring the physical, cellular, and genetic alterations that occur after ethanol exposure1. We then seek to find potential interventions and rapidly test them in this animal model.

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, ...

In vivo Macrophage Imaging Using MR Targeted Contrast Agent for Longitudinal Evaluation of Septic Arthritis

Macrophages are key-cells in the initiation, the development and the regulation of the inflammatory response to bacterial infection. Macrophages are intensively and increasingly recruited in septic joints from the early phases of infection and the infiltration is supposed to regress once efficient removal of the pathogens is obtained. The ability to identify in vivo macrophage activity in an infected joint can therefore provide two main applications: early detection of acute synovitis and monitoring of therapy.
In vivo noninvasive detection of macrophages can be performed with magnetic resonance imaging using iron nanoparticles such as ultrasmall superparamagnetic iron oxide (USPIO). After intravascular or intraarticular administration, USPIO are specifically phagocytized by activated macrophages, and, due to their magnetic properties, induce signal changes in tissues presenting macrophage infiltration. A quantitative evaluation of the infiltrate is feasible, as the area with signal loss (number of dark pixels) observed on gradient echo MR images after particles injection is correlated with the amount of iron within the tissue and therefore reflects the number of USPIO-loaded cells.
We present here a protocol to perform macrophage imaging using USPIO-enhanced MR imaging in an animal model of septic arthritis, allowing an initial and longitudinal in vivo noninvasive evaluation of macrophages infiltration and an assessment of therapy action.

In vivo Macrophage Imaging Using MR Targeted Contrast Agent for Longitudinal Evaluation of Septic Arthritis

We demonstrate how to perform macrophage MR imaging using ultrasmall superparamagnetic contrast agent (USPIO) in septic arthritis, allowing an initial and longitudinal in vivo non-invasive evaluation of macrophages infiltration and an assessment of therapy efficacy.

Macrophages are key-cells in the initiation, the development and the regulation of the inflammatory response to bacterial infection. Macrophages are ...

Computerized Dynamic Posturography for Postural Control Assessment in Patients with Intermittent Claudication

Computerized dynamic posturography with the EquiTest is an objective technique for measuring postural strategies under challenging static and dynamic conditions. As part of a diagnostic assessment, the early detection of postural deficits is important so that appropriate and targeted interventions can be prescribed. The Sensory Organization Test (SOT) on the EquiTest determines an individual&#39;s use of the sensory systems (somatosensory, visual, and vestibular) that are responsible for postural control. Somatosensory and visual input are altered by the calibrated sway-referenced support surface and visual surround, which move in the anterior-posterior direction in response to the individual&#39;s postural sway. This creates a conflicting sensory experience. The Motor Control Test (MCT) challenges postural control by creating unexpected postural disturbances in the form of backwards and forwards translations. The translations are graded in magnitude and the time to recover from the perturbation is computed.
Intermittent claudication, the most common symptom of peripheral arterial disease, is characterized by a cramping pain in the lower limbs and caused by muscle ischemia secondary to reduced blood flow to working muscles during physical exertion. Claudicants often display poor balance, making them susceptible to falls and activity avoidance. The Ankle Brachial Pressure Index (ABPI) is a noninvasive method for indicating the presence of peripheral arterial disease and intermittent claudication, a common symptom in the lower extremities. ABPI is measured as the highest systolic pressure from either the dorsalis pedis or posterior tibial artery divided by the highest brachial artery systolic pressure from either arm. This paper will focus on the use of computerized dynamic posturography in the assessment of balance in claudicants.

Individuals with intermittent claudication exhibit poor balance compared to healthy controls. Computerized dynamic posturography is an objective method for measuring an individual's postural responses to balance disturbances. This provides an objective reflection of the person&#x2019;s ability to respond to situations under which sensory stimuli are altered and unexpected perturbations occur.

Computerized dynamic posturography with the EquiTest is an objective technique for measuring postural strategies under challenging static and dynamic ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

A Methodological Approach to Non-invasive Assessments of Vascular Function and Morphology

The endothelium is the innermost lining of the vasculature and is involved in the maintenance of vascular homeostasis. Damage to the endothelium may predispose the vessel to atherosclerosis and increase the risk for cardiovascular disease. Assessments of peripheral endothelial function are good indicators of early abnormalities in the vascular wall and correlate well with assessments of coronary endothelial function. The present manuscript details the important methodological steps necessary for the assessment of microvascular endothelial function using laser Doppler imaging with iontophoresis, large vessel endothelial function using flow-mediated dilatation, and carotid atherosclerosis using carotid artery ultrasound. A discussion on the methodological considerations for each of the techniques is also presented, and recommendations are made for future research.

The present article describes the methodological considerations for several non-invasive assessments of vascular function and morphology that are commonly used in medical research to assess different stages of atherosclerosis.

The endothelium is the innermost lining of the vasculature and is involved in the maintenance of vascular homeostasis.  Damage to the endothelium may ...

Treating SCA1 Mice with Water-Soluble Compounds to Non-Specifically Boost Mitochondrial Function

Mitochondrial dysfunction plays a significant role in the aging process and in neurodegenerative diseases including several hereditary spinocerebellar ataxias and other movement disorders marked by progressive degeneration of the cerebellum. The goal of this protocol is to assess mitochondrial dysfunction in Spinocerebellar ataxia type 1 (SCA1) and assess the efficacy of pharmacological targeting of metabolic respiration via the water-soluble compound succinic acid to slow disease progression. This approach is applicable to other cerebellar diseases and can be adapted to a host of water-soluble therapies.
Ex vivo analysis of mitochondrial respiration is used to detect and quantify disease-related changes in mitochondrial function. With genetic evidence (unpublished data) and proteomic evidence of mitochondrial dysfunction in the SCA1 mouse model, we evaluate the efficacy of treatment with the water-soluble metabolic booster succinic acid by dissolving this compound directly into the home cage drinking water. The ability of the drug to pass the blood brain barrier can be deduced using high performance liquid chromatography (HPLC). The efficacy of these compounds can then be tested using multiple behavioral paradigms including the accelerating rotarod, balance beam test and footprint analysis. Cytoarchitectural integrity of the cerebellum can be assessed using immunofluorescence assays that detect Purkinje cell nuclei and Purkinje cell dendrites and soma. These methods are robust techniques for determining mitochondrial dysfunction and the efficacy of treatment with water-soluble compounds in cerebellar neurodegenerative disease.

We present a biochemical and behavioral protocol to evaluate the efficacy of mitochondria-targeted water-soluble compounds for the treatment of Spinocerebellar ataxia type 1 (SCA1) and other cerebellar neurodegenerative diseases.

Mitochondrial dysfunction plays a significant role in the aging process and in neurodegenerative diseases including several hereditary spinocerebellar ...

Evaluation of Fluid Overload by Bioelectrical Impedance Vectorial Analysis

Early detection and management of fluid overload are critically important in acute illness, as the impact of therapeutic intervention can result in decreased or increased mortality rates. Accurate fluid status assessment entails appropriate therapy. Unfortunately, as the gold standard method of radioisotopic fluid measurement is costly, time-consuming, and lacks sensitivity in the acute care clinical setting, other less-accurate methods are typically used, such as clinical examination or 24 h output. Bioelectrical impedance vectorial analysis (BIVA) is an alternative impedance-based approach, where the raw parameter resistance and reactance of a subject are plotted to produce a vector, the position of which can be evaluated relative to tolerance intervals in an R-Xc graph. The fluid status is then interpreted as normal or abnormal, based on the distance from the mean vector derived from a healthy reference population. The objective of the present study is to demonstrate how to evaluate the presence of fluid overload through bioelectrical impedance vectorial analysis and the impedance ratio measured with tetrapolar multi-frequency equipment in patients admitted to the emergency department.

In this study, we demonstrate how to evaluate the presence of fluid overload through bioelectrical impedance vectorial analysis (BIVA) and the impedance ratio measured using tetrapolar multi-frequency equipment in patients admitted to the emergency department. BIVA and impedance ratio are reliable and useful tools to predict poor outcomes.

Early detection and management of fluid overload are critically important in acute illness, as the impact of therapeutic intervention can result in ...

Evaluation of Hydration Status by Bioelectrical Impedance Vector Analysis in Patients with Ischemic Heart Disease Undergoing Exercise Stress Test

Ischemic heart disease (IHD) represents a group of clinical syndromes characterized by myocardial ischemia, leading to an impairment in the myocardial blood supply and compromised perfusion. Several clinical variables assessed through a stress test, such as oxygen uptake (VO2) and heart rate oxygen pulse (HR/O2), have been attributed as cardiopulmonary prognostic factors in patients with IHD. However, other factors like hydration status (HS), potentially affecting the cardiopulmonary response, have been barely addressed. Unbalanced HS has a short-term effect on plasma volume and the sympathetic nervous system, which impacts blood volume, and lowers VO2 and HR/O2. Recently, bioelectrical impedance analysis (BIA), a method based on the opposition of body tissues (including fluid volume) to a low electrical current, has been widely used to assess HS by obtaining two components: resistance (R) and reactance (Xc) and using prediction formulas. However, several limitations as chronic illness or abnormal fluid status, may affect the results. In this sense, alternative BIA methods, such as bioelectrical impedance vector analysis (BIVA), have become relevant. R and Xc (adjusted by height) result in a vector plotted on the R/Xc graph, which allows interpreting the HS as normal or abnormal according to the distance of the mean vector. This study aims to describe how to determine HS by BIVA using a single-frequency device and compare the results with the cardiopulmonary response in patients with IHD.

Hydration status imbalances can have a short-term effect on direct and indirect determinants of oxygen uptake and pulse, and morbidity and mortality prognostic factors in ischemic heart disease. This protocol describes the technique for assessment of hydration status through bioelectrical impedance vector analysis and cardiopulmonary response during exercise stress test.

Ischemic heart disease (IHD) represents a group of clinical syndromes characterized by myocardial ischemia, leading to an impairment in the myocardial ...

Evaluation of Changes in Hydration and Body Cell Mass with Bioelectrical Impedance Analysis after Exercise Program for Rheumatoid Arthritis Patients

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