Age-specific association between meal-skipping patterns and the risk of hyperglycemia in Korean adults: a national cross-sectional study using the KNHANES data.

BACKGROUND: Glucose metabolism regulation is influenced by age and meal skipping, although research on their interplay with hyperglycemia remains limited. This study aims to explore the intricate relationship between meal-skipping patterns and hyperglycemia risk across distinct age groups in South Korean adults. METHODS: Utilizing data from the Korea National Health and Nutrition Examination Surveys (KNHANES) conducted from 2013 to 2020, comprising 28,530 individuals aged 19 years and older, this study employed multivariable logistic regression models to examine the associations between meal-skipping patterns and the risk of hyperglycemia. RESULTS: Meal-skipping patterns were categorized into three groups: no skipping (NS), skipping breakfast (SB), and skipping dinner (SD). Age groups were defined as "young" (aged 19-44), "middle-aged" (aged 45-64), and "elderly" adults (over 65 years old). Among "young" adults, SB was associated with a 1.33-fold higher risk of hyperglycemia (OR = 1.33, 95% CI = 1.14-1.54) compared to NS. Conversely, in "elderly" adults, SD was linked to a 0.49-fold reduced risk (95% CI = 0.29-0.82) when compared to NS. Additionally, we observed that the Korean Health Eating Index (KHEI) scores, representing the quality of diet on a scale of 0 to 100, were consistently lower in SB compared to NS across all age groups. Intriguingly, specifically among the "elderly" group, this score was higher in SD compared to NS (p < 0.001). CONCLUSIONS: This study demonstrates age-specific variations in the association between meal-skipping patterns and the risk of hyperglycemia.

Correction: Exploring the Effect of the Dynamics of Behavioral Phenotypes on Health Outcomes in an mHealth Intervention for Childhood Obesity: Longitudinal Observational Study.

[This corrects the article DOI: 10.2196/45407.].

Exploring the Effect of the Dynamics of Behavioral Phenotypes on Health Outcomes in an mHealth Intervention for Childhood Obesity: Longitudinal Observational Study.

BACKGROUND: Advancements in mobile health technologies and machine learning approaches have expanded the framework of behavioral phenotypes in obesity treatment to explore the dynamics of temporal changes. OBJECTIVE: This study aimed to investigate the dynamics of behavioral changes during obesity intervention and identify behavioral phenotypes associated with weight change using a hybrid machine learning approach. METHODS: In total, 88 children and adolescents (ages 8-16 years; 62/88, 71% male) with age- and sex-specific BMI ≥85th percentile participated in the study. Behavioral phenotypes were identified using a hybrid 2-stage procedure based on the temporal dynamics of adherence to the 5 behavioral goals during the intervention. Functional principal component analysis was used to determine behavioral phenotypes by extracting principal component factors from the functional data of each participant. Elastic net regression was used to investigate the association between behavioral phenotypes and weight change. RESULTS: Functional principal component analysis identified 2 distinctive behavioral phenotypes, which were named the high or low adherence level and late or early behavior change. The first phenotype explained 47% to 69% of each factor, whereas the second phenotype explained 11% to 17% of the total behavioral dynamics. High or low adherence level was associated with weight change for adherence to screen time (ß=-.0766, 95% CI -.1245 to -.0312), fruit and vegetable intake (ß=.1770, 95% CI .0642-.2561), exercise (ß=-.0711, 95% CI -.0892 to -.0363), drinking water (ß=-.0203, 95% CI -.0218 to -.0123), and sleep duration. Late or early behavioral changes were significantly associated with weight loss for changes in screen time (ß=.0440, 95% CI .0186-.0550), fruit and vegetable intake (ß=-.1177, 95% CI -.1441 to -.0680), and sleep duration (ß=-.0991, 95% CI -.1254 to -.0597). CONCLUSIONS: Overall level of adherence, or the high or low adherence level, and a gradual improvement or deterioration in health-related behaviors, or the late or early behavior change, were differently associated with weight loss for distinctive obesity-related lifestyle behaviors. A large proportion of health-related behaviors remained stable throughout the intervention, which indicates that health care professionals should closely monitor changes made during the early stages of the intervention. TRIAL REGISTRATION: Clinical Research Information Science KCT0004137; https://tinyurl.com/ytxr83ay.

Sedentary Time and Fast-Food Consumption Associated With Weight Gain During COVID-19 Lockdown in Children and Adolescents With Overweight or Obesity.

BACKGROUND: The coronavirus disease pandemic is predicted to have adverse health effects on children and adolescents who are overweight or obese due to restricted school activity and stay-at-home orders. The purpose of this observational study was to determine the factors associated with weight gain in children and adolescents with overweight and obesity during coronavirus disease 2019 (COVID-19) lockdown. METHODS: Ninety-seven participants (sex- and age-specific body mass index (BMI) ≥ 85th percentile) were included. A baseline examination was conducted pre-COVID-19 (August 2019 to January 2020), and re-examination was performed post-lockdown (June to September 2020) and the results were compared. Correlation and regression analyses were conducted to investigate the association among changes in cardiometabolic markers and lifestyle behaviors with changes in BMI z-score. RESULTS: During the COVID-19 pandemic, an increase in BMI z-score (2.56 [2.01-2.94] to 2.62 [2.03-3.18]) was noticed in children and adolescents with obesity. Changes in cardiometabolic markers including liver enzymes, triglycerides (r = 0.398), leptin (r = 0.578), and adiponectin (r = -0.326), as well as muscular strength (r = -0.212), were correlated with the increase in BMI z-score. According to a multivariate regression analysis, changes in sedentary time (B = 0.016; 95% confidence interval [CI], 0.001-0.032) and fast-food consumption (B = 0.067; 95% CI, 0.013-0.122) were the lifestyle variables associated with BMI z-score increase. CONCLUSION: Changes in lifestyle behaviors including fast-food consumption and sedentary time during the COVID-19 pandemic may be associated with weight gain. In order to prevent health-related risks in children and adolescents with obesity during the pandemic, it is important to maintain the level of physical activity and healthy dietary habits.

Tumor susceptibility gene 101 is required for the maintenance of uterine epithelial cells during embryo implantation.

BACKGROUND: The tumor susceptibility gene 101 (Tsg101), a component of the endosomal sorting complex required for transport (ESCRT) complex I, is involved in multiple biological processes involving endomembranous structures and the plasma membrane. The role of Tsg101 in the uterine epithelium was investigated in Tsg101 floxed mice crossed with Lactoferrin-iCre mice (Tsg101d/d). METHODS: Tsg101d/d mice were bred with stud male mice and the status of pregnancy was examined on days 4 and 6. Histological analyses were performed to examine the uterine architecture. Immunofluorescence staining of several markers was examined by confocal microscopy. Uterine epithelial cells (UECs) were isolated from Tsg101f/f and Tsg101d/d mice, and the expression of necroptosis effectors was examined by RT-PCR, western blotting, and immunofluorescence staining. UECs were also subjected to RNA expression profiling. RESULTS: Tsg101d/d female mice were subfertile with implantation failure, showing unattached blastocysts on day 6 of pregnancy. Histological and marker analyses revealed that some Tsg101d/d day 4 pregnant uteri showed a disintegrated uterine epithelial structure. Tsg101d/d UECs began to degenerate within 18 h of culture. In UECs, expression of necroptosis effectors, such as RIPK1, RIPK3, and MLKL were first confirmed. UECs responded to a stimulus to activate necroptosis and showed increased cell death. CONCLUSIONS: Tsg101 deficiency in the uterine epithelium causes implantation failure, which may be caused by epithelial defects. This study provides evidence that UECs harbor a necroptotic machinery that responds to death-inducing signals. Thus, Tsg101 expression in the uterine epithelium is required for normal pregnancy in mice.

Perivascular Stem Cell-Derived Cyclophilin A Improves Uterine Environment with Asherman's Syndrome via HIF1α-Dependent Angiogenesis.

Asherman's syndrome (AS) is characterized by intrauterine adhesions or fibrosis resulting from scarring inside the endometrium. AS is associated with infertility, recurrent miscarriage, and placental abnormalities. Although mesenchymal stem cells show therapeutic promise for the treatment of AS, the molecular mechanisms underlying its pathophysiology remain unclear. We ascertained that mice with AS, like human patients with AS, suffer from extensive fibrosis, oligo/amenorrhea, and infertility. Human perivascular stem cells (hPVSCs) from umbilical cords repaired uterine damage in mice with AS, regardless of their delivery routes. In mice with AS, embryo implantation is aberrantly deferred, which leads to intrauterine growth restriction followed by no delivery at term. hPVSC administration significantly improved implantation defects and subsequent poor pregnancy outcomes via hypoxia inducible factor 1α (HIF1α)-dependent angiogenesis in a dose-dependent manner. Pharmacologic inhibition of HIF1α activity hindered hPVSC actions on pregnancy outcomes, whereas stabilization of HIF1α activity facilitated such actions. Furthermore, therapeutic effects of hPVSCs were not observed in uterine-specific HIF1α-knockout mice with AS. Secretome analyses of hPVSCs identified cyclophilin-A as the major paracrine factor for hPVSC therapy via HIF1α-dependent angiogenesis. Collectively, we demonstrate that hPVSCs-derived cyclophilin-A facilitates HIF1α-dependent angiogenesis to ameliorate compromised uterine environments in mice with AS, representing the major pathophysiologic features of humans with AS.

Relationship Between Bone Mineral Density and Body Composition According to Obesity Status in Children.

OBJECTIVE: To analyze the relationship between body composition, metabolic parameters, and bone mineral density (BMD) according to sex and the degree of obesity in children and adolescents. METHODS: A total of 236 subjects with obesity, aged 10 to 15 years (36.9% girls), were enrolled. Obesity was classified into simple (SmOb) and extreme (ExOb) forms. The BMD of the total body, less head, was measured by dual energy x-ray absorptiometry, and the BMD z-score was used to evaluate the relationship of body composition with metabolic parameters. RESULTS: BMD z-scores were higher in subjects with ExOb than in those with SmOb. Lean mass index (LMI), body mass index z-score, and vitamin D intake showed positive relationships, whereas percentage of body fat and serum leptin level showed negative relationships with BMD z-scores in boys. In girls, LMI and body mass index z-score showed positive relationships with BMD z-scores. In multivariable linear regressions, serum leptin level showed negative relationships with BMD z-score, only in boys. In addition, positive relationships of LMI and negative relationships of percentage of body fat with BMD z-scores were observed in subjects with SmOb. However, positive relationships of LMI with BMD z-scores were attenuated in subjects with ExOb. CONCLUSION: High BMD appears to be positively associated with lean mass in children and adolescents with obesity, which might be a natural protective mechanism to withstand the excess weight. However, excessive body fat appears to be negatively associated with BMD, which might attenuate the positive relationship between lean mass and BMD in subjects with ExOb.

Lipidomic changes in mouse oocytes vitrified in PEG 8000-supplemented vitrification solutions.

Cryopreserved oocytes are inevitably exposed to cold stress, which negatively affects the cellular aspects of the oocytes. Lipidomic analysis of the oocytes reveals quantitative changes in lipid classes under conditions of cold stress, leading to potential freezing-vulnerability. We had previously shown that specific phospholipids are significantly downregulated in vitrified-warmed mouse oocytes compared to those in fresh oocytes. In this study, we examined whether supplementation of polyethylene glycol 8000 (PEG 8000) during vitrification influences the lipidome of the oocytes. We used liquid chromatography with tandem mass spectrometry (LC-MS/MS) to study the alteration in the lipidome in three groups of mouse oocytes: fresh, vitrified-warmed, and vitrified with PEG 8000-warmed during vitrification. In these groups, we targeted to analyze 21 lipid classes. We profiled 132 lipid species in the oocytes and statistical analyses revealed lipid classes that were up- or downregulated in these groups. Overall, our data revealed that several classes of lipids were affected during vitrification, and that oocytes vitrified with PEG 8000 to some extent alleviated the levels of changes in phospholipid and sphingolipid contents during vitrification. These results suggest that phospholipids and sphingolipids are influenced by PEG 8000 during vitrification and that PEG 8000 can be considered as a potential candidate for preserving membrane integrity during oocyte cryopreservation.

Molecular analysis of lipid uptake- and necroptosis-associated factor expression in vitrified-warmed mouse oocytes.

BACKGROUND: We had previously demonstrated that vitrification reduces the levels of certain phospholipid classes, and that oocytes from aged mice show a similar lipidome alteration, even without vitrification. In the current investigation, we examined if vitrification-warming of mouse oocytes from young and aged mice causes any changes in molecular aspects of lipid-associated features. METHODS: Metaphase II (MII) stage oocytes were harvested from young (10-14-week-old) and aged (45-54-week-old) mice by a superovulation regime with PMSG followed by hCG. We examined the status of the intracellular lipid pool and the integrity of the plasma membrane by staining oocytes with BODIPY 500/510 and CellMask live dyes. Expression of lipid uptake- and necroptosis-associated genes was assessed by quantitative PCR analyses, in oocytes from young and old mice, before and after vitrification. Localization patterns of two crucial necroptosis proteins, phosphorylated MLKL (pMLKL) and phosphorylated RIPK1 (pRIPK1) were examined in mouse oocytes by immunofluorescence staining. Necrostain-1 (Nec1), an inhibitor of RIPK1, was used to examine if RIPK1 activity is required to maintain oocyte quality during vitrification. RESULTS: We confirmed that vitrified-warmed oocytes from aged mice showed noticeable decrease in both CellMask and BODIPY 500/510 dyes. Among the lipid uptake-associated genes, Cd36 expression was higher in oocytes from aged mice. Necroptosis is a type of programmed cell death that involves damage to the plasma membrane, eventually resulting in cell rupture. The expression of necroptosis-associated genes did not significantly differ among groups. We observed that localization patterns of pMLKL and pRIPK1 were unique in mouse oocytes, showing association with microtubule organizing centers (MTOCs) and spindle poles. pMLKL was also localized on kinetochores of MII chromosomes. Oocytes treated with Nec1 during vitrification showed a decreased survival rate, indicating the importance of RIPK1 activity in oocyte vitrification. CONCLUSIONS: We report that oocytes from aged mice show differential expression of CD36, which suggests that CD36-mediated lipid uptake may be influenced by age. We also show for the first time that pMLKL and pRIPK1 exhibit unique localization pattern in mouse oocytes and this may suggest role(s) for these factors in non-necroptosis-associated cellular processes.

Sarcopenia in relation to nutrition and lifestyle factors among middle-aged and older Korean adults with obesity.

PURPOSE: To examine the prevalence of sarcopenic obesity (SO) and its association with nutrition and lifestyle factors. METHODS: Data from the 2008-2011 Korea National Health and Nutrition Examination Survey for 3937 Korean individuals aged 40 years or older with obesity defined by the modified Asia-Pacific criteria (BMI ≥ 25 kg/m2) were used. Sarcopenia was defined as an appendicular skeletal muscle mass divided by weight (%) of < 1 SD below the sex-specific mean for young adults. Logistic regression models were used for the associations controlling covariates. RESULTS: The prevalence of SO was 52.5%. The SO group had insufficient energy intake, protein, and antioxidant micronutrients, a lower overall dietary quality, a lower physical activity, and higher rates of negative psychological factors than the non-sarcopenic obesity group (p < 0.05). After controlling for covariates, having high mean adequacy ratio scores which was calculated by averaging the sum of the nutrient adequacy ratios had a 94% lower prevalence of SO in the older individuals. Low participation in aerobic exercises had a 74% higher prevalence of SO in the older aged group, and high participation in flexibility exercises had a 11% lower prevalence of SO in the middle-aged group. Interestingly, negative psychological factors were found only in the middle-aged adults with SO (p < 0.05). CONCLUSION: Sarcopenia was present in over half of the middle-aged and older Korean adults with obesity. Having high dietary quality, increased physical activity, and positive psychological health were associated with a low prevalence of SO.

Association between combinations of nutritional status and quality of life and food purchasing motives among the elderly in South Korea.

BACKGROUND: In the elderly, nutritional status and quality of life (QOL) could potentially affect food purchasing behaviors. We examined the association between combinations of nutritional status and QOL and food purchasing motives among the elderly. METHODS: A total of 143 community-dwelling elderly were recruited in Seoul, South Korea. Nutritional status and QOL were assessed and participants were divided into four groups according to those combinations. Binary logistic regression analysis was used to examine the odds of food purchasing motives according to combinations of nutritional status and QOL. RESULTS: As a result of comparing the scores (mean ± SD) of the overall important factors for food purchasing, health related factors such as Nutrition quality and Preventive of treatment effect were the highest score (4.4 ± 0.8), followed by Price (4.1 ± 0.9), Ease of purchase (3.8 ± 0.9), Ease of chewing (3.7 ± 0.9), and Taste (3.6 ± 0.9). Participants with a low nutritional status and low QOL had more eating-related problems (77.8%) including chewing difficulty (48.9%) and constipation (17.8%) than those with a high nutritional status and high QOL (P < 0.05). Participants who were in high nutritional status and low QOL were more likely to be motivated by Ease of chewing (OR: 6.72; 95% CI: 1.44-31.37; P < 0.05), while those who were in low nutritional status and high QOL were less motivated by Taste (OR: 0.28; 95% CI: 0.08-0.94; P < 0.05) compared to those who were in high nutritional status and high QOL. CONCLUSIONS: There were differences in food purchasing motives such as Ease of chewing or Taste according to combinations of nutritional status and QOL. These data are important in demonstrating differing motives for food choice across nutritional status and QOL, and also provide indications of which care service and food development may be needed in promoting health for the elderly in South Korea.

A significantly enhanced antibacterial spectrum of D-enantiomeric lipopeptide bactenecin.

Cationic antimicrobial peptides (CAMPs) are important antibiotics because they possess a broad spectrum of activity against both Gram-positive and Gram-negative bacteria, including those resistant to traditional antibiotics. The cyclic peptide bactenecin is a 12-amino acid CAMP that contains one intramolecular disulfide bond. To improve the antibacterial activity of bactenecin, we designed and synthesized several bactenecin analogs by applying multiple approaches, including amino acid substitution, use of the d-enantiomeric form, and lipidation. Among the synthetic analogs, d-enantiomeric bactenecin conjugated to capric acid, which we named dBacK-(cap), exhibited a significantly enhanced antibacterial spectrum with MIC values ranging from 1 to 8 µM against both Gram-positive and Gram-negative bacteria, including some drug-resistant bacteria. Upon exposure to dBacK-(cap), S. aureus cells were killed within 1 h at the MIC value, but full inactivation of E. coli required over 2 h. These results indicate that covalent addition of a d-amino acid and a fatty acid to bactenecin is the most effective approach for enhancing its antibacterial activity.

Estrogen induces EGR1 to fine-tune its actions on uterine epithelium by controlling PR signaling for successful embryo implantation.

The harmonized actions of ovarian E2 and progesterone (P4) regulate the proliferation and differentiation of uterine cells in a spatiotemporal manner. Imbalances between these hormones often lead to infertility and gynecologic diseases. Whereas numerous factors that are involved in P4 signaling have been identified, few local factors that mediate E2 actions in the uterus have been revealed. Here, we demonstrate that estrogen induces the transcription factor, early growth response 1 ( Egr1), to fine-tune its actions in uterine epithelial cells (ECs) that are responsible for uterine receptivity for embryo implantation. In the presence of exogenous gonadotrophins, ovulation, fertilization, and embryonic development normally occur in Egr1-/- mice, but these animals experience the complete failure of embryo implantation with reduced artificial decidualization. Although serum levels of E2 and P4 were comparable between Egr1+/+ and Egr1-/- mice on d 4 of pregnancy, aberrantly reduced levels of progesterone receptor in Egr1-/- uterine ECs caused enhanced E2 activity and impaired P4 response. Ultrastructural analyses revealed that Egr1-/- ECs are not fully able to provide proper uterine receptivity. Uterine mRNA landscapes in Egr1-/- mice revealed that EGR1 controls the expression of a subset of E2-regulated genes. In addition, P4 signaling was unable to modulate estrogen actions, including those that are involved in cell-cycle progression, in ECs that were deficient in EGR1. Furthermore, primary coculture of Egr1-/- ECs with Egr1+/+ stromal cells, and vice versa, supported the notion that Egr1 is required to modulate E2 actions on ECs to prepare the uterine environment for embryo implantation. In contrast to its role in ECs, loss of Egr1 in stroma significantly reduced stromal cell proliferation. Collectively, our results demonstrate that E2 induces EGR1 to streamline its actions for the preparation of uterine receptivity for embryo implantation in mice.-Kim, H.-R., Kim, Y. S., Yoon, J. A., Yang, S. C., Park, M., Seol, D.-W., Lyu, S. W., Jun, J. H., Lim, H. J., Lee, D. R., Song, H. Estrogen induces EGR1 to fine-tune its actions on uterine epithelium by controlling PR signaling for successful embryo implantation.

Glycaemic indices and glycaemic loads of common Korean carbohydrate-rich foods.

Glycaemic index (GI) and glycaemic load (GL) values of foods consumed in Asia remain poorly characterised despite the fact that Asian diets are high in carbohydrates. We evaluated the GI and GL of the most commonly consumed carbohydrate-rich foods, according to food type and cooking methods. GI and GL values were determined using protocols from the FAO/WHO and International Standards Organization recommendations. A total of 152 healthy subjects were enrolled in the study. In all, forty-nine carbohydrate-rich foods were categorised as cereal grains, noodles and pasta, breads and other processed grains and starchy vegetables, prepared using standard cooking methods and evaluated. Cereal grains had the widest range of GI values that the food made with white rice and barley had GI values of 51-93 and 35-70, respectively, according to cooking methods, and most cereal grains had high GL values. Noodles and pasta had low to medium GI values, but most foods had high GL values. Breads had medium to high GI and GL values, while other processed grains had low to medium GI and GL values. The GI values for food made with starchy vegetables (e.g. potatoes and sweet potatoes) varied widely for different cooking methods but tended to have low GL values. In conclusion, GI values for a single food type varied widely with the cooking method used. This study of GI and GL values for common carbohydrate-rich foods provides a valuable reference for consumers and health professionals to make informed food choices for glycaemic control.

Validation of body composition using bioelectrical impedance analysis in children according to the degree of obesity.

This study aimed to validate body composition analysis using bioelectrical impedance analysis (BIA) against dual-energy X-ray absorptiometry (DXA) in children with obesity and to compare agreement between BIA and DXA according to their degree of obesity. Three hundred and sixteen children aged 6-17 years participated in the Intervention for Childhood and Adolescents Obesity via Activity and Nutrition study. We divided participants by body mass index (BMI) percentile (group 1: mild to moderate obesity; group 2: severe obesity) and compared body composition variables, eg, percentage of body fat (%BF), fat mass (FM), and fat-free mass (FFM) using BIA and DXA. The %BF and FM of BIA were significantly lower (-1.8% and -0.8 kg, respectively), and the FFM of BIA was significantly higher (1.4 kg) than those of DXA. There were significant negative relationships between the absolute value of differences from BIA and DXA and BMI z-scores in %BF, FM, and FFM (regression coefficient [ß]: -1.39, 95% confidence interval [CI]: -1.81 to -0.97; ß: -0.34, 95%CI: -0.61 to -0.06; ß: -0.73, 95%CI: -1.03 to -0.44, respectively). The gap of body compositions between BIA and DXA decreased as participants became more obese, and the differences of FM in boys with severe obesity and the differences of %BF and FFM in girls with severe obesity were much less than those in children with mild to moderate obesity. In conclusion, the agreement between DXA and BIA was better for children with severe obesity than for children with mild to moderate obesity.

Expression of Egr3 in mouse gonads and its localization and function in oocytes.

OBJECTIVE: The early growth response (Egr) family consists of four members (Egr1, Egr2, Egr3, and Egr4) that are zinc finger transcription factors. Among them, Egr3 is involved in transcriptional regulation of target genes during muscle spindle formation and neurite outgrowth. We previously showed that the immunoreactive Egr3 is localized on oocyte spindle and accumulate near the microtubule organizing center during meiosis I in mice. Egr3 was also shown to be localized on spermatocytes. We herein investigated if Egr3 is expressed in mouse gonads and if Egr3 blockade results in any defect in oocyte maturation. METHODS: Expression of Egr3 in mouse gonads was examined by reverse transcription-polymerase chain reaction. Full-length Egr3 and truncated Egr3 (ΔEgr3) complementary RNAs (cRNAs) with Xpress tag at N-terminus and DsRed2 at C-terminus, and small interfering RNA (siRNA) targeting Egr3 were microinjected into mouse oocytes at germinal vesicle stage. Localization of microinjected Egr3 was examined by confocal live imaging and immunofluorescence staining. RESULTS: Egr3 mRNA was detected in mouse ovaries and testes from 1 to 4 week-old mice. An uncharacterized longer transcript containing 5'untranslated region was also detected in 3 and 4 week-old gonads. Microinjected Xpress-Egr3-DsRed2 or Xpress-ΔEgr3-DsRed2 localized to nuclei and chromosomes during meiotic progression. Microinjection of these cRNAs or Egr3 siRNA in oocytes did not affect meiotic maturation. Immunofluorescence staining of Egr3 in Xpress-ΔEgr3-DsRed2-injected oocytes showed a positive signal only on meiotic spindle, suggesting that this antibody does not detect endogenous or exogenous Egr3 in mouse oocytes. CONCLUSION: The results show that Egr3 localizes to chromosomes during meiotic progression and that certain antibodies may not faithfully represent localization of target proteins in oocytes. Egr3 seems to be dispensable during oocyte maturation in mice.

Rapamycin Influences the Efficiency of In vitro Fertilization and Development in the Mouse: A Role for Autophagic Activation.

The mammalian target of rapamycin (mTOR) regulates cellular processes such as cell growth, metabolism, transcription, translation, and autophagy. Rapamycin is a selective inhibitor of mTOR, and induces autophagy in various systems. Autophagy contributes to clearance and recycling of macromolecules and organelles in response to stress. We previously reported that vitrified-warmed mouse oocytes show acute increases in autophagy during warming, and suggested that it is a natural response to cold stress. In this follow-up study, we examined whether the modulation of autophagy influences survival, fertilization, and developmental rates of vitrified-warmed mouse oocytes. We used rapamycin to enhance autophagy in metaphase II (MII) oocytes before and after vitrification. The oocytes were then subjected to in vitro fertilization (IVF). The fertilization and developmental rates of vitrified-warmed oocytes after rapamycin treatment were significantly lower than those for control groups. Modulation of autophagy with rapamycin treatment shows that rapamycin-induced autophagy exerts a negative influence on fertilization and development of vitrified-warmed oocytes.

Exosomal proteins in the aqueous humor as novel biomarkers in patients with neovascular age-related macular degeneration.

Age-related macular degeneration (AMD) describes the progressive degeneration of the retinal pigment epithelium (RPE), retina, and choriocapillaris and is the leading cause of blindness in people over 50. The molecular mechanisms underlying this multifactorial disease remain largely unknown. To uncover novel secretory biomarkers related to the pathogenesis of AMD, we adopted an integrated approach to compare the proteins identified in the conditioned medium (CM) of cultured RPE cells and the exosomes derived from CM and from the aqueous humor (AH) of AMD patients by LC-ESI-MS/MS. Finally, LC-MRM was performed on the AH from patients and controls, which revealed that cathepsin D, cytokeratin 8, and four other proteins increased in the AH of AMD patients. The present study has identified potential biomarkers and therapeutic targets for AMD treatment, such as proteins related to the autophagy-lysosomal pathway and epithelial-mesenchymal transition, and demonstrated a novel and effective approach to identifying AMD-associated proteins that might be secreted by RPE in vivo in the form of exosomes. The proteomics-based characterization of this multifactorial disease could help to match a particular marker to particular target-based therapy in AMD patients with various phenotypes.

Autophagic activation in vitrified-warmed mouse oocytes.

Vitrification involves the use of cryoprotectants (CPAs) and liquid nitrogen (LN2), which may cause osmotic damage and cryoinjury to oocytes. Autophagy is widely recognized as a survival or response mechanism elicited by various environmental and cellular stressors. However, the induction of autophagy in vitrified-warmed oocytes has not been examined. In this work, we investigated whether the vitrification-warming process induces autophagy in mouse oocytes. Metaphase II (MII) oocytes that were vitrified and stored in LN2 for at least 2 weeks were used in the study. In RT-PCR analyses, we observed that several Atg genes such as Atg5, Atg7, Atg12, LC3a (Map1lc3a), LC3b (Map1lc3b), and Beclin1 were expressed in MII mouse oocytes. Slight reduction in mRNA levels of Atg7 and Atg12 in vitrified-warmed oocytes was noted, and expression of these genes was not significantly influenced. Confocal live imaging analysis using oocytes from GFP-LC3 transgenic mice revealed that vitrified-warmed oocytes had a significantly higher number of GFP-LC3 puncta in comparison to fresh oocytes. The expression of BECLIN1 protein was also increased in vitrified-warmed oocytes. Treatment with 3-methyladenine, an inhibitor of autophagy, did not significantly affect the rates of oocyte survival, IVF, and embryonic development after warming and IVF. The results suggest that the observed autophagic activation in vitrified-warmed oocytes is a natural adaptive response to cold stress. Collectively, we show for the first time that vitrified-warmed mouse oocytes exhibit autophagic activation during warming and that this response is not induced by CPA-containing solutions. The induction of autophagy by cold temperature is first reported herein.

Association between obesity and metabolic co-morbidities among children and adolescents in South Korea based on national data.

BACKGROUND: Childhood obesity has become a serious public health threat worldwide due to its many short- and long-term adverse health effects. We assessed the association between weight status and metabolic co-morbidities among South Korean children using nationally representative data. METHODS: Data from the 2007-2008 Korea National Health and Nutrition Examination Surveys for 1,526 children aged 10-19 years were used. Logistic regression models were fit to examine the association between weight status [overweight, 85th percentile≤BMI<95th percentile; obese, BMI≥95th percentile; and central obesity, waist circumference≥90th percentile, based on 2007 Korean National Growth Charts] and metabolic outcomes. RESULTS: More obese children had metabolic co-morbidities than normal-weight children (P<0.05). Boys had higher means BMI than girls, WC, and BP, while girls had higher means of total blood cholesterol and HDL-cholesterol (P<0.05). Attributable risk of metabolic syndrome was high due to overweight and obesity: 91.1% for central obesity and 29.6% for high TG. Obese children had increased risk of having high BP (adjusted odds ratio (OR): 1.90; 95% CI: 1.05-3.45), dyslipidemia (OR: 6.21; 95% CI: 3.59-10.75), high TG (OR: 6.87; 95% CI: 4.05-11.64), low HDL (OR: 4.46; 95% CI: 2.23-8.89), and ≥2 co-morbidities (OR: 26.97; 95% CI: 14.95-48.65) compared to normal-weight subjects, while the associations between weight status and metabolic outcomes were stronger in boys. CONCLUSIONS: Obesity was strongly associated with metabolic co-morbidities in South Korean children.