Aplio 400

The CEUS liver parenchymal phase was used to determine the phagocytic capacity of Kupffer cells31 (link). The detailed protocol for imaging the liver through contrast ultrasonography has been published elsewhere32 (link). This time, we applied this protocol with some modification for the current CEUS measurements. Briefly, sonazoid used as a contrast agent was diluted to 0.1 mL/1200 g/body weight and injected into the vein of the subjects. A clinical gastroenterologist scanned the liver of the subjects using 1-second intermittent transmission scans at 40 min using ultrasonography [Aplio 400, Toshiba medical, Tokyo, Japan]. The fluorescent ROI [intensity in the region of interest] was calculated using the equipment software Advanced Dynamic Flow [Toshiba medical, Tokyo, Japan].

All study participants had undergone transthoracic echocardiography as part of periodic pre-participation evaluation (PPE) due to innocent heart murmurs or suspicion of abnormal electrocardiographic findings. The studies were performed at the National Centre for Sports Medicine between 2013 and 2017. Children thus found to have significant acquired or congenital heart diseases affecting normal heart size and haemodynamics were excluded. Echocardiograms were performed by two experienced sonographers using a commercially available ultrasound scanner (Toshiba Aplio 400, Toshiba Medical Systems Europe, Zoetermeer, the Netherlands), according to recent guidelines [15 (link)]. All measurements were taken in 2-dimensional parasternal long-axis view (PLAX) in end-diastole. The measurements were taken from inner edge to inner edge and reported to within 1 mm. Persons exhibiting ambiguous results of any measurement were excluded from the initial study group. LV mass (in grams) was calculated using the Devereux equation: LVM = 0.8{1.04[(LV cavity dimension + posterior wall thickness + interventricular septal thickness)³ − (LV cavity dimension)³]} + 0.6 [9 (link)].

Both the right and left carotid arteries were evaluated from the longitudinal and transverse views of the common (CCA), bifurcation, and internal carotid arteries (ICA) using a high-resolution B-mode ultrasonography (Aplio 400, Toshiba, Japan). Intima-media thickness (IMT) was measured in triplicate using a computer-assisted method by experienced sonographers blinded to the clinical and laboratory data. Mean IMT was defined as the average of the mean values from the distal 1 cm of the far walls of both right and left CCA. Plaque was defined as a focal wall thickening of ≥1.5 mm or ≥50% of the surrounding IMT [11 (link)]. The severity of plaque was evaluated as a plaque score which was calculated as the sum of points (range 0 to 12) of all 6 segments. In each segment, 1 point per plaque was allocated for the near and far walls of each segment (CCA, bifurcation, and ICA) of the right and left carotid arteries [12 (link)].

A renal ultrasound of the right kidney was performed in seven FGR and six control neonates within 24 hours after birth (Aplio 400, Toshiba medical systems, Zoetermeer, The Netherlands, 7 MHz, convex probe). Pulsatility and resistance index in renal artery and renal length were measured 2–3 times and means were subsequently calculated. Renal ultrasound was not conducted in all patients due to unavailability of investigators and in one case due to poor neonatal condition. Most ultrasounds were performed by the same investigator (DCV) with exception of two ultrasounds.

All patients underwent preoperative thyroid US, which was performed by 2 radiologists with 4 and 13 years of experience in performing thyroid US examination. High-resolution ultrasound scanners (iU 22, Phillips Medical Systems, Bothell, WA, USA; and Aplio 400, Toshiba Medical Systems, Tokyo, Japan), with 5–12 MHz and 8–15 MHz linear probes, respectively, were used. One of the 2 US instruments was arbitrarily used for each patient.