A high prevalence of heavy menstrual bleeding (HMB) has been reported in women with Fontan circulation. Cyanosis has been reported to contribute to HMB, and menstruation has been suggested to affect cardiac status in women with congenital heart disease. This study aimed to evaluate the relationship between the amount of menstrual flow and cardiac status in women with Fontan circulation.Twenty women who had undergone the Fontan procedure were prospectively investigated and the amount of their menstrual flow was evaluated using a questionnaire. Participants were divided into two groups-small and large menstrual bleeding groups-and their clinical data, including the results of hematological tests and echocardiographic findings, were evaluated.One (5%) woman showed primary amenorrhea. Eight of the remaining 19 (42%) women were included in the large menstrual bleeding group. Women with large menstrual bleeding showed a significantly higher hematocrit level (47.1% [36.2%-50.3%] versus 42.1% [35.3%-44.9%], P = 0.006) and longer QRS duration (106 [92-172] ms versus 88 [78-140] ms, P = 0.008), as well as a lower fractional area change (37.4% [35.6%-47.2%] versus 47.0% [38.2%-55.7%], P = 0.010) and global longitudinal strain (−10.5% [−14.9% to −6.6%] versus −13.9% [−20.5% to −7.8%], P = 0.041) of the dominant ventricle on echocardiography, than women with small bleeding.Erythrocytosis, longer QRS duration, and reduced ventricular function were related to increased menstrual bleeding in women with Fontan circulation. These functions may be interrelated with the amount of menstrual flow in such women.

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Tricuspid annular plane systolic excursion (TAPSE), tricuspid lateral annular systolic velocity, fractional area change (FAC), and right ventricular (RV) free wall longitudinal strain are indices of RV function measured by transthoracic echocardiography [1]. The current guideline of the American Heart Association and American Thoracic Society recommend these echocardiographic indices as important follow-up tools for assessing RV function in patients with pulmonary hypertension [2]. However, this guideline is mainly based on data from patients with pulmonary arterial hypertension [2,3]. A few previous reports have shown that mean pulmonary artery pressure (PAP) plays an important role in evaluating the severity and prognosis of pulmonary diseases such as chronic obstructive pulmonary disease, idiopathic pulmonary fibrosis, and sarcoidosis [4,5]. Although right heart failure is known to be progressive in patients with pulmonary diseases, few studies have examined the relationship between echocardiographic indices of RV function and the severity or patient prognosis of pulmonary diseases.

IntroductionFormation of cell spheres is an important procedure in biomedical research. A large number of high-quality cell spheres of uniform size and shape are required for basic studies and therapeutic applications. Conventional approaches, including the hanging drop method and suspension culture, are used for cell sphere production. However, these methods are time consuming, cell spheres cannot be harvested easily, and it is difficult to control the size and geometry of cell spheres. To resolve these problems, a novel multiple-funnel cell culture insert was designed for size controlling, easy harvesting, and scale-up production of cell spheres.MethodsThe culture substrate has 680 micro-funnels with a 1-mm width top, 0.89 mm depth, and 0.5 mm square bottom. Mouse embryonic stem cells were used to test the newly developed device. The seeded embryonic stem cells settled at the downward medium surface toward the bottom opening and aggregated as embryoid bodies (EBs). For cell sphere harvest, the bottom of the culture insert was put in contact with the medium surface in another culture dish, and the medium in the device flowed down with cell spheres by hydrostatic pressure.ResultsCompact cell spheres with uniform size and shape were collected easily. The diameter of the spheres could be controlled by adjusting the seeding cell density. Spontaneous neural differentiation (nestin and Tju1) and retinoic acid-induced endodermal differentiation (Pdx-1 and insulin I) were improved in the EBs produced using the new insert compared to those in EBs produced by suspension culture.ConclusionsThis novel cell culture insert shall improve future studies of cell spheres and benefit clinical applications of cell therapy.