An Alternative Approach to Determining Metabolic Syndrome Component Cutoffs in Children and Adolescents Using Segmental Regression Analysis.

Background: The prevalence of metabolic syndrome is increasing in children and adolescents. Although some diagnostic criteria for metabolic syndrome exist, further research is needed to determine appropriate age-, sex-, and race-specific cutoffs for each component. Methods and Results: Health examinations were conducted in 1,679 children aged 6-15 years in 9 regions of Japan. Participants were divided into 3 age groups for each sex: 6-8, 9-11, and 12-15 years. For metabolic syndrome components in each group, inverse cumulative percentile graphs were drawn and approximated by 3 regression lines using segmented regression analysis. The intersection of each regression line was defined as the breakpoint, and the measured value corresponding to the breakpoint percentile as the breakpoint value. Breakpoint values for waist circumference were age dependent at approximately 60, 70, and 80 cm for ages 6-8, 9-11, and 12-15 years, respectively. Breakpoint values for blood pressure were age- and/or sex dependent, while those for triglycerides, high-density lipoprotein cholesterol, and fasting blood glucose were neither age nor sex dependent. Based on these results, we proposed new cutoffs for diagnosing metabolic syndrome in Japanese children and adolescents. Conclusions: Breakpoint values obtained by segmented regression analysis on inverse cumulative percentile graphs can be useful for determining metabolic syndrome component cutoffs in children and adolescents.

Developmental trajectories at a high risk for childhood overweight/obesity.

BACKGROUND: The associations between developmental patterns (trajectories) in children and maternal factors have been widely investigated, but paternal effects on these trajectories are unclear. This study aimed to determine child and parental factors involved in developmental trajectories at high risk for causing adverse cardiovascular (CV) profiles in children. METHODS: We analyzed longitudinal anthropometric data from birth to the present and CV profiles of 1,832 healthy volunteers (51% girls) aged 3-15 years who participated in a nationwide study between July 2012 and January 2014. Six trajectory latent class growth models were developed using body mass index z- scores. Predictors for being in developmental trajectories at high risk for causing adverse CV profiles were determined by multivariate regression analysis. RESULTS: The mean±standard deviation number of anthropometric data points was 12±3 for both boys and girls. Among the six trajectories, the infantile onset and continual increase groups had significantly worse levels of many CV profiles than those in the remaining groups. Paternal overweight/obesity was an independent predictor for boys being in the infantile onset group and for girls being in the continual increase group. Additionally, maternal pre-pregnancy overweight/obesity in boys and maternal excessive gestational weight gain in girls were independent predictors for being in the infantile onset group. Having no sibling in boys and an older maternal age were independent predictors for being in the continual increase group. CONCLUSIONS: Interventions to prevent childhood obesity should include strategies that focus on fathers and mothers as well as those that focus on children with certain types of familial background.

Promoting physical activity through walking to treat childhood obesity, mainly for mild to moderate obesity.

BACKGROUND: There are no randomized controlled trials examining the effect of walking on childhood obesity. METHODS: A randomized controlled trial was conducted between August 2014 and April 2015 in Japan. Elementary school children aged 6 to 12 years with a percentage overweight (%OW) of ≥20% were recruited. One hundred and ninety children wanted to participate in the program, and all were accepted. After viewing a video that promoted physical activity through walking, participants were randomly assigned to three groups: walking (≥10 000 steps on school holidays), limiting screen time (<90 min on weekdays and <150 min on school holidays), and a control group (no intervention). The primary outcome was a decrease in %OW after 3 months' intervention. Per protocol analysis was performed using 156 participants who fulfilled the inclusion criteria of a %OW ≥20%. RESULTS: The mean %OW was 35 ± 7% before intervention. The mean reduction in %OW after intervention in the walking (n = 59), limiting ST (n = 46), and control (n = 51) groups were -4.06 ± 4.84, -1.97 ± 4.62, and -1.81 ± 3.64 percentage points, respectively. Reduction in %OW was significantly larger in the walking group than in the control group: adjusted mean difference, -2.18 percentage points (95% confidence interval, -3.85 to -0.52), P = 0.002. The intervention in children also had favorable effects on the lifestyles of their parents. The intention-to-treat analysis of all 190 participants showed comparable results. CONCLUSION: Promoting physical activity through walking on school holidays may be an additional strategy for treating elementary school children with obesity.

Transplantation of Human Corneal Endothelial Cells Cultured on Bio-Engineered Collagen Vitrigel in a Rabbit Model of Corneal Endothelial Dysfunction.

PURPOSE: To evaluate the effect of transplanting bioengineered corneal endothelial grafts in a rabbit model of corneal endothelial failure. METHODS: Human corneal endothelial cells (HCECs) were seeded on a vitrigel carrier. After Descemet's membrane was removed from the eyes of rabbits, transplantation was done with a vitrigel/HCEC graft or vitrigel alone without cells, or the eyes were left untreated. Slit lamp examinations and measurement of the central corneal thickness (CCT) were performed for 14 days postoperatively. RESULTS: HCECs cultured on vitrigel were strongly positive for ZO-1 and Na+/K+ ATPase. On day 14, the cornea showed mild edema and the pupil margins were visible through the grafts in the vitrigel/HCEC graft group. HCECs completely covered the grafts on day 14. In contrast, there was severe corneal edema and the pupil margins were undetectable on day 14 after transplantation of the vitrigel carrier alone or no transplantation. Proliferation of host cells was not observed in these groups. On day 14, the mean CCT was significantly thinner in the vitrigel/HCEC graft group than in the other two groups (p = 0.0008). CONCLUSIONS: Transplantation of a vitrigel/HCEC graft was effective for reducing the corneal thickness and restoring corneal transparency, suggesting the usefulness of vitrigel as a carrier for corneal endothelial cells.

High-density collagen patch prevents stricture after endoscopic circumferential submucosal dissection of the esophagus: a porcine model.

BACKGROUND AND AIMS: Extensive excision of the esophageal mucosa by endoscopic submucosal dissection (ESD) frequently evokes a luminal stricture. This study aimed to determine the efficacy of a high-density collagen patch for the prevention of esophageal stricture in extensive ESD. METHODS: Six pigs underwent circumferential esophageal ESD under general anesthesia. In 3 pigs, artificial ulcers were covered by 2 collagen patches. The other 3 pigs underwent circumferential ESD only. RESULTS: The 2 collagen patches were settled onto the ulcer surface using a general endoscope and instruments. The collagen patch-treated group showed significantly better patency rates on both the oral and anal sides of the wound area compared with the control group at day 14. The mucosal re-epithelization ratio was significantly promoted, and the extent of mucosal inflammation and fibrosis was significantly decreased with the collagen patch treatment in the wound area. The frequency of cells positive α-smooth muscle actin was significantly reduced in the collagen patch-treated group compared with the control group. CONCLUSIONS: We have established a high-density collagen device that can reduce the esophageal stricture associated with extensive ESD. This easy-to-handle device would be useful during superficial esophageal cancer treatment by ESD.

Non-high-density lipoprotein cholesterol as a cardiovascular risk screening tool in children.

BACKGROUND: Non-high-density lipoprotein cholesterol (non-HDL-C) is now recognized as a strong predictive factor for cardiovascular disease in adults, but there have been few reports on non-HDL-C in children. METHODS: A total of 5853 4th and 7th grade schoolchildren were included in the screening for lifestyle-related disease from 2010 to 2011 in Takaoka City. The children underwent anthropometric measurements and non-fasting blood tests to measure total cholesterol (TC), HDL-C, triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C). The relationship between percent overweight and each lipid level was analyzed, and children above the 97th percentile level with respect to both TC (220 mg/dL) and non-HDL-C (152 mg/dL) were compared and assessed. The relationship between non-HDL-C and the prevalence of metabolic syndrome was also analyzed among 150 obese children. RESULTS: Non-HDL-C positively correlated with percent overweight and TG rather than did TC (r = 0.273, 0.360 vs 0.118, 0.179, all P < 0.001). In the screening using TC criteria, several subjects with increased HDL-C were miscategorized as hyperlipidemic, whereas none were miscategorized using the non-HDL-C criteria. The sensitivity of the identification of increased LDL-C was lower when the criteria for TC were used rather than the criteria for non-HDL-C (80.8% vs 98.3%). Among obese children, the prevalence of metabolic syndrome increased significantly and was accompanied by an increase in non-HDL-C (P = 0.009). CONCLUSION: Instead of TC, non-HDL-C would serve as a better and useful cardiovascular risk screening tool for lifestyle-related disease in school children.

Development of an oxygenation culture method for activating the liver-specific functions of HepG2 cells utilizing a collagen vitrigel membrane chamber.

We recently developed a collagen vitrigel membrane (CVM) chamber possessing a scaffold composed of high-density collagen fibrils. In this study, we first confirmed that the advantage of CVM chamber in comparison to the traditional culture chamber with porous polyethylene terephthalate membrane is to preserve a culture medium poured in its inside even though the under side is not a liquid phase but solid and gas phases. Subsequently, we designed three different culture systems to grow HepG2 cells in a culture medium (liquid phase) on the CVM which the under side is a culture medium, a plastic surface (solid phase) or 5 % CO2 in air (gas phase) and aimed to develop a brief culture method useful for activating the liver-specific functions and analyzing the pharmacokinetics of fluorescein diacetate. HepG2 cells cultured for 2 days on the liquid-solid interface and subsequently for 1 day on the liquid-gas interface represented excellent cell viability and morphology in comparison to the others, and remarkably improved albumin secretion and urea synthesis to almost the same level of freshly isolated human hepatocytes and CYP3A4 activity to about half the level of differentiated HepaRG cells. Also, the cells rapidly absorbed fluorescein diacetate, distributed it in cytosol, metabolized it into fluorescein, and speedily excreted fluorescein into both bile canaliculus-like networks and extracellular solution. These data suggest that hepatic structure and functions of monolayered HepG2 cells can be induced within a day after the oxygenation from beneath the CVM.

A new cell-free bandage-type artificial skin for cutaneous wounds.

Engineered skin substitutes are widely used in skin wound management. However, no currently available products satisfy all the criteria of usability in emergency situations, easy handling, and minimal scar formation. To overcome these shortcomings, we designed a cell-free bandage-type artificial skin, named "VitriBand" (VB), using adhesive film dressing, silicone-coated polyethylene terephthalate film, and collagen xerogel membrane defined as a dried collagen vitrigel membrane without free water. We analyzed its advantages over in-line products by comparing VB with hydrocolloid dressing and collagen sponge. For evaluation, mice inflicted with full-thickness skin defects were treated with VB, hydrocolloid dressing, and collagen sponge. A plastic film group treated only with adhesive film dressing and silicone-coated polyethylene terephthalate film, and a no treatment group were also compared. VB promoted epithelization while inhibiting the emergence of myofibroblasts and inflammation in the regenerating tissue more effectively than the plastic film, hydrocolloid dressing, and collagen sponge products. We have succeeded in establishing a cell-free bandage-type artificial skin that could serve as a promising first-line medical biomaterial for emergency treatment of skin injuries in various medical situations.

Development and evaluation of porcine atelocollagen vitrigel membrane with a spherical curve and transplantable artificial corneal endothelial grafts.

PURPOSE: To develop a collagen vitrigel (CV) optimized as a corneal endothelial cell (CEC) carrier and create an artificial corneal endothelial graft. METHODS: We first developed a flat-shaped collagen vitrigel for regenerative medicine (CV-RM) using porcine atelocollagen and ultraviolet (UV) irradiation. The optimal UV amount was determined by measuring the CV-RM transparency under various irradiating conditions. The collagen vitrigel for corneal endothelial regenerative treatment (CV-CERT), a transparent porcine atelocollagen with a curved shape, was made using spherically curved molds and UV irradiation. The membrane permeability of the CV-CERT was tested in vitro. The biocompatibility, transparency, and adhesiveness of the CV-CERT were evaluated in rabbit eyes. We also developed a culture technique for distributing human CECs on the curved CV-CERT. RESULTS: The optimal amount of UV irradiation for CV-RM transparency was 2400 mJ/cm(2). Membrane permeability of CV-CERT at day 5 was higher than that of commercially available CV (P = 0.032). The CV-CERT was transparent and biocompatible in rabbit corneas for up to 4 months. The CV-CERT remained attached to the rabbit corneal posterior surface, whereas the flat-shaped CV-RM, differing only in shape from the CV-CERT, dislocated soon after surgery. Human CECs seeded on the CV-CERT using our technique were evenly distributed with a single layer structure and a mean cell density of 2650 ± 100 cells/mm(2). CONCLUSIONS: We developed a transparent and biocompatible porcine-derived atelocollagen vitrigel membrane with a spherical curvature. A transplantable artificial endothelial graft was created by combining cultured human CECs and the CV-CERT.

Epithelial-to-mesenchymal transition and slit function of mesothelial cells are regulated by the cross talk between mesothelial cells and endothelial cells.

Peritoneal dysfunction is a major factor leading to treatment failure of peritoneal dialysis (PD). However, the precise mechanism of the peritoneal diffusion changes related to PD remains to be elucidated. To this end, we have established a novel peritoneal diffusion model in vitro, which consists of a three-dimensional culture system using a collagen vitrigel membrane chamber and a fluid-stream generation system. This artificial peritoneal model revealed that high-glucose culture medium and fluid flow stress promoted the epithelial-mesenchymal transition (EMT) process of mesothelial cells and that endothelial cells inhibited this mesothelial EMT process. Mesothelial cells in the EMT state showed high expression of connective tissue growth factor and low expression of bone morphogenic protein-7, while non-EMT mesothelial cells showed the opposite expression pattern of these two proteins. In addition, these protein expressions were dependent on the presence of endothelial cells in the model. Our model revealed that the endothelial slit function was predominantly dependent on the covering surface area, while the mesothelial layer possessed a specific barrier function for small solutes independently of the surface area. Notably, a synergic barrier effect of mesothelial cells and endothelial cells was present with low-glucose pretreatment, but high-glucose pretreatment abolished this synergic effect. These findings suggest that the mesothelial slit function is not only regulated by the high-glucose-induced EMT process but is also affected by an endothelial paracrine effect. This peritoneal diffusion model could be a promising tool for the development of PD.

Impact of lifestyles of adolescents and their parents on cardiovascular risk factors in adolescents.

AIM: The aim of this study was to determine the impact of the lifestyles of adolescents and their parents on the levels of cardiovascular (CV) risk factors in the adolescents. METHODS: A total of 755 volunteers (331 male, 424 female) aged 15 to 18 years were included. Abdominal obesity, hypertension, elevated triglyceride levels, decreased high density lipoprotein-cholesterol levels, and hyperglycemia were considered to be CV risk factors. Self-reported lifestyle, including participation in school-based extracurricular (EC) physical activities, time spent on physical activity or watching television (TV), and average daily food intake were assessed. Parental information on weight status and lifestyle was also obtained. RESULTS: Multivariate regression analyses showed that participation in EC physical activities, time spent watching TV, regular breakfast consumption, total energy intake, fiber intake per 1,000 Kcal, and parental BMI were independently associated with the levels of one or more CV risk factors in adolescents. Among these, participation in EC physical activities had a profound effect on adolescent CV risk factor levels. The risk of male adolescent obesity was associated with paternal obesity, but not with maternal obesity. Conversely, the risk of female adolescent obesity was associated with maternal obesity but not with paternal obesity. CONCLUSIONS: Participation in EC physical activities may be the first-line approach for adolescents to maintain favorable CV risk factor levels. An association between paternal or maternal obesity and adolescent obesity differs between adolescent genders in Japan; thus, approaches focusing on parents should take the gender of the adolescent into consideration.

Impact of having one cardiovascular risk factor on other cardiovascular risk factor levels in adolescents.

AIM: Little is known about the impact of having one cardiovascular (CV) risk factor on the levels of other CV risk factors in the general adolescent population. We hypothesized that when adolescents have one CV risk factor, the levels of other CV risk factors worsen simultaneously. METHODS: Subjects consisted of 1,257 healthy adolescent volunteers (549 males and 708 females) aged 15-18 years. Abdominal obesity, hypertension, raised triglyceride levels, decreased HDL-cholesterol levels and hyperglycemia were used as CV risk factors. Homeostasis assessment of insulin resistance (HOMA-IR) was used as a surrogate marker of insulin resistance. Levels of four biomarkers, leptin, adiponectin, high-sensitive C-reactive protein, and desacyl-ghrelin, were also determined. Cut-offs for gender-specific individual CV risk factor levels were based on the 90th (or 10th) percentile values of the subjects in the present study. RESULTS: The levels of all CV risk factors and HOMA-IR significantly and simultaneously worsened when adolescents had one CV risk factor in both genders. Having any one CV risk factor indicated the development of other CV risk factors in adolescents; in particular, we found that the development of abdominal obesity in male subjects had a harmful effect on the levels of other CV risk factors and was associated with the worsening of all four biomarkers examined. CONCLUSIONS: It is important to determine the presence or absence of these CV risk factors before and/or during adolescence, because having one CV risk factor indicates the start of an accumulation of CV risk factors in the general adolescent population.