IJCS | Volume 31, Nº3, May/ June 2018

276 Figure 1 - Urine collection schedule on day without therapeutic intervention (Control) and on day with manual lymphatic drainage (MLD). Camargo et al. Manual lymphatic drainage, natriuresis and lipolysis International Journal of Cardiovascular Sciences. 2018;31(3)274-281 Original Article In the lower limbs, drainage of inguinal lymph nodes was started by 10 strokes to evacuate the lymph. With the hands contacting the skin, a pressure was put to promote the flow of the lymph towards internal iliac lymph nodes, followed by 10 strokes in each region, from proximal to distal thigh to direct the flow of the lymph to the internal saphenous vein. Ten strokes were performed on the knees, aimed at cleaning popliteal lymph nodes, by dividing the area in two regions (upper and lower). Then, drainage of legs and thighs was performed, promoting lymph to drain toward the anterointernal part of the leg. In the ankles, 10 strokes in the retromalleolar regionwere made, directing the lymph to the leg, whichwas repeated on the feet. At the end of the procedure, a clearingmotion was performed from the foot to the upper part of the leg, followed by 10 motions on popliteal lymph nodes and one motion toward upper thigh. The procedure was finished by 10 strokes on inguinal lymph nodes. Urine collection and assessment of urine composition Urine was collected at four-time points with 60-minute intervals – -60, 0, 60 and 120minutes. Urine samples were collected using 1000mL beakers in awater closet available by the experiment room. Volunteers were instructed to completely empty the bladder for correct assessment of urine flow. The sample collected at -60min aimed at excluding potential influence of dietary or climate factors on the results. This protocol was followed on two days – one (control) day without therapeutic intervention and one day of MLD, performed between urine collection at 0 and 60 minutes 11 (Figure 1). Urinary excretion of the following compounds was calculated from urinary flow rate (mL/min): sodium, by titration with silver nitrate (mM/min), glycerol (mmol/min) by colorimetric assay (LABORCLIN, SP); and ANP (pg/min) by ELISA (USCNLife Science Inc., Houston, EUA). Statistical analysis S t a t i s t i ca l ana l ys i s wa s pe r f ormed us i ng GraphPadPrism 5.0 software (Inc, La Jolla, CA, USA). Normal distribution was tested by the Shapiro-Wilk test. Data without normal distribution were expressed as median and interquartile range (25%-75%), whereas those with normal distribution as mean ± standard error. For data without normal distribution, theMann-Whitney test andWilcoxon test were used for unpaired (different “n”) and paired (same “n”) data, respectively, and normally distributed data were analyzed by the unpaired t-test. Statistical significance was set at 5%. Results In within-group analysis, on both days (control and MLD), initial values (0 min) in the nOCPu group showed increased urinary flow rate without changes in sodiumor glycerol excretion. The same analysis showed that there were no statistical differences in OCPu between days. However, between-group comparison showed that, at the beginning of the study, urinary flow rate on both days and sodium excretion on control day were lower in OCPu than nOCPu (Table 1).