Nitrate sources, timing, and pathways of a permeable volcanic aquifer system with mixed land use in Jeju Island, South Korea.

The occurrence of various N-related human activities increases the difficulty in distinguishing the major sources of NO3- contamination in groundwater, especially in areas with mixed land uses. In addition, the estimation of the timing and pathways of NO3- is necessary to better understand the processes of NO3- contamination in the subsurface aquifer system. This study applied environmental tracers, such as stable isotopes and age tracers (δ15N and δ18O of NO3-, δ11B, chlorofluorocarbons, and 3H), to elucidate the sources, timing, and pathways of NO3- contamination in the groundwaters of the Hanrim area, which has suffered from illegal disposal of livestock wastes since the 1980s, and also characterizes them based on mixed N-contaminant sources such as chemical fertilizers and sewage. The combined use of δ15N and δ11B overcame the limitation of using only NO3- isotopes for the identification of overlapping sources of N and successfully identified the major source of N as livestock wastes. The lumped parameter model (LPM) estimated the binary mixing of the young (age: 23-40 years, NO3-N: 2.55-15.10 mg/L) and old (age: >60 years, NO3-N: <3 mg/L) groundwaters, and explained their age mixing behaviors. The young groundwater was highly affected by livestock-derived N loading during 1987-1998, which coincides with the period of improper dumping of livestock wastes. Furthermore, the young groundwater with elevated NO3-N followed the historical NO3-N curves with younger ages (6 and 16 years) than those derived from the LPM, suggesting the possibility of faster inflows of livestock wastes through the permeable volcanic structures. This study demonstrated that a comprehensive understanding of NO3- contamination processes can be achieved using environmental tracer methods, which enables the efficient management of groundwater resources in areas with multiple N sources.

Impact of abdominal obesity on the risk of glioma development in patients with diabetes: A nationwide population-based cohort study in Korea.

BACKGROUND: Abdominal obesity has been suggested as a risk factor for glioma; however, it is unclear whether this association applies to people with diabetes. This study examined the association between abdominal obesity and the risk of developing gliomas in diabetic patients. METHODS: We conducted a retrospective cohort study using the National Health Insurance System of South Korea from 2009 to 2012. The primary outcome was the incidence of newly diagnosed gliomas according to waist circumference (WC), and subgroup analyses were performed according to demographic characteristics and diabetes status including disease duration, number of oral hypoglycemic agents, and insulin use. RESULTS: Of a total of 1,893,057 participants, 1,846 (0.10%) cases of gliomas occurred. After adjusting for confounding factors, WC ≥90 cm (men)/85 cm (women) was associated with significantly higher risks of gliomas (adjusted HR [95% CI]; 1.279 [1.053, 1.554], 1.317 [1.048, 1.655], and 1.369 [1.037, 1.807] in the WC <95 cm (men)/90 cm (women) group, WC <100 cm (men)/95 cm (women) group, and WC ≥100 cm (men)/95 cm (women) group, respectively). Subgroup analysis showed that patients with larger WC had a consistently higher incidence of glioma than their lean counterparts, except for insulin users (insulin user vs. nonuser, P for interaction = .03). CONCLUSIONS: Abdominal obesity was associated with the development of gliomas in diabetic patients in a nationwide population-based database. Further study is needed in diabetic patients to stratify the risk for glioma development according to WC and to establish the underlying mechanism of carcinogenesis.

Integrated application of a Bayesian mixing model, numerical model, and environmental tracers to characterize groundwater recharge sources in a mountainous area.

In this study, the combined use of a Bayesian mixing model (BMM), numerical model (random walk particle tracking-RWPT), and environmental tracers (δ18O-δD, 3H, and CFC) was applied to elucidate the probabilistic contribution of the recharge sources, flow path, and residence time of groundwater across the mountainous area of Jeju Island, South Korea. Especially, the BMM ability to estimate the variable recharge contributions to the aquifer by different elevations and seasons was investigated. The δ18O-δD isotopes showed that groundwater in the study area was primarily fed by precipitation during the wet season, and the BMM estimated that wet season recharge contributed to approximately 64% of the total. The BMM-based probabilistic estimation of recharge sources revealed a mixed contribution of source waters from different elevations. A notable difference in recharge flow path was observed between highland (>450 masl) and lowland (<400 masl) wells, where the inflow of source water from the regional flow was dominant in the former and both regional and local recharges served as significant groundwater sources in the latter. Evidence from age tracers (3H and CFC-12) also supported different recharge mechanisms between highland and lowland wells. A reasonable match between the BMM- and RWPT-derived recharge contributions (RMSE 0.02-0.06) was achieved within the uncertainty ranges, with RWPT being particularly useful for capturing different flow paths between highland and lowland wells. The dynamics revealed here provide important information for establishing an improved and informed groundwater management plan for the mountainous area of Jeju Island. Ultimately, this study highlights the advantageous integrated analysis of BMM, RWPT, and environmental tracer analyses to enhance the reliability of recharge area estimation and increase the collective understanding of complex hydrogeological systems in mountainous areas.

Association between organochlorine pesticides and nonalcoholic fatty liver disease in the National Health and Nutrition Examination Survey 2003-2004.

While endocrine disruptors are emerging as a cause of nonalcoholic fatty liver disease (NAFLD), little is known about the link between NAFLD and organochlorine pesticides (OCPs), one of the endocrine disruptors. We retrospectively analyzed the U.S. National Health and Nutrition Examination Survey 2003-2004 and compared the baseline demographics in individuals according to the presence of NAFLD (fatty liver index [FLI] ≥ 60). Logistic regression analysis was performed to determine whether OCP concentration affected NAFLD prevalence and subgroup analyses regarding NAFLD-related variables and advanced hepatic fibrosis (FIB-4 ≥ 2.67) were performed. Of the 1515 individuals, 579 (38.2%) had NAFLD. Oxychlordane showed concentration-dependent risk for NAFLD (OR 3.471 in fourth quartile [Q4]; 95% CI 1.865-6.458; P = 0.007). p,p'-DDE and trans-nonachlor showed similar trends without statistical significance. Conversely, mirex showed the lowest risk for NAFLD in the highest concentration quartile (OR 0.29 in Q4; 95% CI 0.175-0.483; P < 0.001). Oxychlordane showed the most pronounced association with the levels of each component of FLI and liver enzymes. None of the OCPs were significantly associated with advanced fibrosis. In conclusion, among OCPs, exposure to oxychlordane showed the most prominent impact associated with NAFLD.

Sphingosine 1-Phosphate Receptor 4 Promotes Nonalcoholic Steatohepatitis by Activating NLRP3 Inflammasome.

BACKGROUND & AIMS: Sphingosine 1-phosphate receptors (S1PRs) are a group of G-protein-coupled receptors that confer a broad range of functional effects in chronic inflammatory and metabolic diseases. S1PRs also may mediate the development of nonalcoholic steatohepatitis (NASH), but the specific subtypes involved and the mechanism of action are unclear. METHODS: We investigated which type of S1PR isoforms is activated in various murine models of NASH. The mechanism of action of S1PR4 was examined in hepatic macrophages isolated from high-fat, high-cholesterol diet (HFHCD)-fed mice. We developed a selective S1PR4 functional antagonist by screening the fingolimod (2-amino-2-[2-(4- n -octylphenyl)ethyl]-1,3- propanediol hydrochloride)-like sphingolipid-focused library. RESULTS: The livers of various mouse models of NASH as well as hepatic macrophages showed high expression of S1pr4. Moreover, in a cohort of NASH patients, expression of S1PR4 was 6-fold higher than those of healthy controls. S1pr4+/- mice were protected from HFHCD-induced NASH and hepatic fibrosis without changes in steatosis. S1pr4 depletion in hepatic macrophages inhibited lipopolysaccharide-mediated Ca++ release and deactivated the Nod-like receptor pyrin domain-containning protein 3 (NLRP3) inflammasome. S1P increased the expression of S1pr4 in hepatic macrophages and activated NLRP3 inflammasome through inositol trisphosphate/inositol trisphosphate-receptor-dependent [Ca++] signaling. To further clarify the biological function of S1PR4, we developed SLB736, a novel selective functional antagonist of SIPR4. Similar to S1pr4+/- mice, administration of SLB736 to HFHCD-fed mice prevented the development of NASH and hepatic fibrosis, but not steatosis, by deactivating the NLRP3 inflammasome. CONCLUSIONS: S1PR4 may be a new therapeutic target for NASH that mediates the activation of NLRP3 inflammasome in hepatic macrophages.

Hepatic MIR20B promotes nonalcoholic fatty liver disease by suppressing PPARA.

Background: Non-alcoholic fatty liver disease (NAFLD) is characterized by excessive lipid accumulation and imbalances in lipid metabolism in the liver. Although nuclear receptors (NRs) play a crucial role in hepatic lipid metabolism, the underlying mechanisms of NR regulation in NAFLD remain largely unclear. Methods: Using network analysis and RNA-seq to determine the correlation between NRs and microRNA in human NAFLD patients, we revealed that MIR20B specifically targets PPARA. MIR20B mimic and anti-MIR20B were administered to human HepG2 and Huh-7 cells and mouse primary hepatocytes as well as high-fat diet (HFD)- or methionine-deficient diet (MCD)-fed mice to verify the specific function of MIR20B in NAFLD. We tested the inhibition of the therapeutic effect of a PPARα agonist, fenofibrate, by Mir20b and the synergic effect of combination of fenofibrate with anti-Mir20b in NAFLD mouse model. Results: We revealed that MIR20B specifically targets PPARA through miRNA regulatory network analysis of nuclear receptor genes in NAFLD. The expression of MIR20B was upregulated in free fatty acid (FA)-treated hepatocytes and the livers of both obesity-induced mice and NAFLD patients. Overexpression of MIR20B significantly increased hepatic lipid accumulation and triglyceride levels. Furthermore, MIR20B significantly reduced FA oxidation and mitochondrial biogenesis by targeting PPARA. In Mir20b-introduced mice, the effect of fenofibrate to ameliorate hepatic steatosis was significantly suppressed. Finally, inhibition of Mir20b significantly increased FA oxidation and uptake, resulting in improved insulin sensitivity and a decrease in NAFLD progression. Moreover, combination of fenofibrate and anti-Mir20b exhibited the synergic effect on improvement of NAFLD in MCD-fed mice. Conclusions: Taken together, our results demonstrate that the novel MIR20B targets PPARA, plays a significant role in hepatic lipid metabolism, and present an opportunity for the development of novel therapeutics for NAFLD. Funding: This research was funded by Korea Mouse Phenotyping Project (2016M3A9D5A01952411), the National Research Foundation of Korea (NRF) grant funded by the Korea government (2020R1F1A1061267, 2018R1A5A1024340, NRF-2021R1I1A2041463, 2020R1I1A1A01074940, 2016M3C9A394589324), and the Future-leading Project Research Fund (1.210034.01) of UNIST.

Perilipin 5 is a novel target of nuclear receptor LRH-1 to regulate hepatic triglycerides metabolism.

Liver receptor homolog-1 (LRH-1) has emerged as a regulator of hepatic glucose, bile acid, and mitochondrial metabolism. However, the functional mechanism underlying the effect of LRH-1 on lipid mobilization has not been addressed. This study investigated the regulatory function of LRH-1 in lipid metabolism in maintaining a normal liver physiological state during fasting. The Lrh-1f/f and LRH-1 liver-specific knockout (Lrh-1LKO) mice were either fed or fasted for 24 h, and the liver and serum were isolated. The livers were used for qPCR, western blot, and histological analysis. Primary hepatocytes were isolated for immunocytochemistry assessments of lipids. During fasting, the Lrh-1LKO mice showed increased accumulation of triglycerides in the liver compared to that in Lrh-1f/f mice. Interestingly, in the Lrh-1LKO liver, decreases in perilipin 5 (PLIN5) expression and genes involved in ß-oxidation were observed. In addition, the LRH-1 agonist dialauroylphosphatidylcholine also enhanced PLIN5 expression in human cultured HepG2 cells. To identify new target genes of LRH-1, these findings directed us to analyze the Plin5 promoter sequence, which revealed -1620/-1614 to be a putative binding site for LRH-1. This was confirmed by promoter activity and chromatin immunoprecipitation assays. Additionally, fasted Lrh-1f/f primary hepatocytes showed increased co-localization of PLIN5 in lipid droplets (LDs) compared to that in fasted Lrh-1LKO primary hepatocytes. Overall, these findings suggest that PLIN5 might be a novel target of LRH-1 to mobilize LDs, protect the liver from lipid overload, and manage the cellular needs during fasting. [BMB Reports 2021; 54(9): 476-481].

Delineation of groundwater quality locations suitable for target end-use purposes through deep neural network models.

Groundwater is the main source of water for beverages, and its quality varies depending on extraction location; this is particularly the case in regions with complex geology, topography, and multiple forms of land use. Thus, it is important to determine a suitable groundwater extraction location based on intended water use and the related water quality standards. In this study, deep neural network (DNN) models and GIS data relating to groundwater quality were applied to estimate potential maps of Gangwon Province in South Korea, where groundwater is frequently extracted for drinking purposes. These maps specify areas where the groundwater quality is conducive for being used as mineral water and water for brewing coffee (hereafter referred as "coffee water"). Sensitivity analysis identified how inputs were sensitive to model estimation and showed that land-use variables were the most sensitive. The importance of each variable quantified how good or bad its region is for the desired groundwater. The overall features of importance were similar between mineral water and coffee water. However, with differences in hydrogeological units, carbonate rock was a variable of high positive importance for mineral water; metamorphic rock was its equivalent for coffee water. Our results offer a potential map of desired groundwater quality in the absence of a detailed understanding of the underlying hydrochemical processes governing groundwater quality. Additionally, the development of such a potential mapping model can help to determine the appropriate development area of groundwater for their respective purposes.

Differentiation of natural and anthropogenic contaminant sources using isotopic and microbial signatures in a heavily cultivated coastal area.

Hydrogeochemical and multiple isotope data for groundwater samples were obtained and interpreted to discriminate anthropogenic and natural contaminant sources in a coastal aquifer underlying a heavily cultivated watershed in Hwaseong, South Korea. The local aquifers are vulnerable to contamination, due to high anthropogenic N inputs and the location close to the ocean facilitating seawater intrusion. Thus, to effectively control the groundwater quality in the study area, it is necessary to differentiate between anthropogenic and natural contaminant sources. The concentrations of NO3-N in the groundwater ranged between 0.14 and 45.6 mg/L in August 2015 and 0.2-39.6 mg/L in March 2016. High concentrations of Cl- (388-1107 mg/L) and a high electrical conductivity (1027-2715 µS/cm) were observed in the study area, suggesting that the groundwater was affected by seawater intrusion. Furthermore, δ15N-NO3-, δ34S-SO42- values and 87Sr/86Sr of groundwater were determined to reveal the origins of the natural and anthropogenic contaminants and the groundwater mean residence times (MRT) and 87Sr/86Sr ratios were used to assess the hydrogeochemical processes along the flow path in the study area. Young groundwater was affected by an anthropogenic contamination source with contributions of 26-46% adding nitrate to the aquifer, whereas old groundwater was impacted by mixing with seawater with contributions of 10-20% with low concentrations of NO3-N, but elevated concentrations of chloride and sulfate. Recently recharged uncontaminated groundwater showed oxic conditions with a diverse microbial community structure, whereas young groundwater contaminated by anthropogenic sources showed a less diverse microbial community structure. The results of this study suggest that multiple isotopes combined with groundwater MRT and microbial data can be applied to distinguish natural and anthropogenic contaminant sources in a groundwater system.

Insulin Resistance Increases Serum Immunoglobulin E Sensitization in Premenopausal Women.

Background: Although studies have shown that obesity is associated with aeroallergen sensitization (atopy), controversy still exists. We aimed to investigate the association between metabolic status, obesity, and atopy stratified by sex and menopausal status. Methods: A total of 1,700 adults from the 2010 Korean National Health and Nutrition Examination Survey were classified into metabolically healthy nonobese (MHNO), metabolically unhealthy nonobese (MUNO), metabolically healthy obese (MHO), and metabolically unhealthy obese (MUO) by body mass index and insulin resistance. Atopy was defined as a positive response to at least one aeroallergen. Multiple regression analysis was used to evaluate the risk of immunoglobulin E (IgE) elevation or atopy in relation to the degree of metabolic abnormality and obesity. Results: In premenopausal women, total IgE was positively correlated with obesity and insulin resistance. MUNO participants had a higher risk of having elevated total IgE compared to MHNO participants (odds ratio [OR], 2.271; 95% confidence interval [CI], 1.201 to 4.294), while MHO participants did not show a significant difference (OR, 1.435; 95% CI, 0.656 to 3.137) in premenopausal women. MUNO, but not MHO was also associated with atopy (OR, 2.157; 95% CI, 1.284 to 3.625). In men and postmenopausal women, there was no significant difference between metabolic status, obesity, and atopy among groups. Conclusion: Increased insulin resistance is associated with total IgE and atopy in premenopausal women but not in postmenopausal women or men.

Mitophagy deficiency increases NLRP3 to induce brown fat dysfunction in mice.

Although macroautophagy/autophagy deficiency causes degenerative diseases, the deletion of essential autophagy genes in adipocytes paradoxically reduces body weight. Brown adipose tissue (BAT) plays an important role in body weight regulation and metabolic control. However, the key cellular mechanisms that maintain BAT function remain poorly understood. in this study, we showed that global or brown adipocyte-specific deletion of pink1, a Parkinson disease-related gene involved in selective mitochondrial autophagy (mitophagy), induced BAT dysfunction, and obesity-prone type in mice. Defective mitochondrial function is among the upstream signals that activate the NLRP3 inflammasome. NLRP3 was induced in brown adipocyte precursors (BAPs) from pink1 knockout (KO) mice. Unexpectedly, NLRP3 induction did not induce canonical inflammasome activity. Instead, NLRP3 induction led to the differentiation of pink1 KO BAPs into white-like adipocytes by increasing the expression of white adipocyte-specific genes and repressing the expression of brown adipocyte-specific genes. nlrp3 deletion in pink1 knockout mice reversed BAT dysfunction. Conversely, adipose tissue-specific atg7 KO mice showed significantly lower expression of Nlrp3 in their BAT. Overall, our data suggest that the role of mitophagy is different from general autophagy in regulating adipose tissue and whole-body energy metabolism. Our results uncovered a new mitochondria-NLRP3 pathway that induces BAT dysfunction. The ability of the nlrp3 knockouts to rescue BAT dysfunction suggests the transcriptional function of NLRP3 as an unexpected, but a quite specific therapeutic target for obesity-related metabolic diseases.Abbreviations: ACTB: actin, beta; BAPs: brown adipocyte precursors; BAT: brown adipose tissue; BMDMs: bone marrow-derived macrophages; CASP1: caspase 1; CEBPA: CCAAT/enhancer binding protein (C/EBP), alpha; ChIP: chromatin immunoprecipitation; EE: energy expenditure; HFD: high-fat diet; IL1B: interleukin 1 beta; ITT: insulin tolerance test; KO: knockout; LPS: lipopolysaccharide; NLRP3: NLR family, pyrin domain containing 3; PINK1: PTEN induced putative kinase 1; PRKN: parkin RBR E3 ubiquitin protein ligase; RD: regular diet; ROS: reactive oxygen species; RT: room temperature; UCP1: uncoupling protein 1 (mitochondrial, proton carrier); WT: wild-type.

Sphingomyelin synthase 1 mediates hepatocyte pyroptosis to trigger non-alcoholic steatohepatitis.

OBJECTIVE: Lipotoxic hepatocyte injury is a primary event in non-alcoholic steatohepatitis (NASH), but the mechanisms of lipotoxicity are not fully defined. Sphingolipids and free cholesterol (FC) mediate hepatocyte injury, but their link in NASH has not been explored. We examined the role of free cholesterol and sphingomyelin synthases (SMSs) that generate sphingomyelin (SM) and diacylglycerol (DAG) in hepatocyte pyroptosis, a specific form of programmed cell death associated with inflammasome activation, and NASH. DESIGN: Wild-type C57BL/6J mice were fed a high fat and high cholesterol diet (HFHCD) to induce NASH. Hepatic SMS1 and SMS2 expressions were examined in various mouse models including HFHCD-fed mice and patients with NASH. Pyroptosis was estimated by the generation of the gasdermin-D N-terminal fragment. NASH susceptibility and pyroptosis were examined following knockdown of SMS1, protein kinase Cδ (PKCδ), or the NLR family CARD domain-containing protein 4 (NLRC4). RESULTS: HFHCD increased the hepatic levels of SM and DAG while decreasing the level of phosphatidylcholine. Hepatic expression of Sms1 but not Sms2 was higher in mouse models and patients with NASH. FC in hepatocytes induced Sms1 expression, and Sms1 knockdown prevented HFHCD-induced NASH. DAG produced by SMS1 activated PKCδ and NLRC4 inflammasome to induce hepatocyte pyroptosis. Depletion of Nlrc4 prevented hepatocyte pyroptosis and the development of NASH. Conditioned media from pyroptotic hepatocytes activated the NOD-like receptor family pyrin domain containing 3 inflammasome (NLRP3) in Kupffer cells, but Nlrp3 knockout mice were not protected against HFHCD-induced hepatocyte pyroptosis. CONCLUSION: SMS1 mediates hepatocyte pyroptosis through a novel DAG-PKCδ-NLRC4 axis and holds promise as a therapeutic target for NASH.

Application of geographically weighted regression models to predict spatial characteristics of nitrate contamination: Implications for an effective groundwater management strategy.

Groundwater nitrate contamination has been the main water quality problem threatening the sustainable utilization of water resources in Jeju Island, South Korea. The spatially varying distribution of nitrate levels associated with complex environmental and anthropogenic factors has been a major challenge restricting improved groundwater management. In this study, we applied ordinary least squares (OLS) regression and geographically weighted regression (GWR) models to determine the relationships between the NO3-N concentration and various parameters (topography, hydrology and land use) across the island. A comparison between the OLS regression and GWR prediction models showed that the GWR models outperformed the OLS regression models, with a higher R2 and a lower corrected Akaike Information Criterion (AICc) value than the OLS regression models. Interestingly, the GWR model was able to provide undiscovered information that was not revealed in the OLS regression models. For example, the GWR model found that orchards (OR) and urban (UR) variables significantly contributed to nitrate enrichment in the certain parts of the island, whereas these variables were ignored as a statistically insignificant factor in the OLS regression model. Our study highlighted that GWR models are a useful tool for investigating spatially varying relationships between groundwater quality and environmental factors; therefore, it can be applied to establish advanced groundwater management plans by reflecting the spatial heterogeneity associated with environmental and anthropogenic conditions.

Autophagic flux defect in diabetic kidney disease results in megamitochondria formation in podocytes.

Podocyte injury is an important factor in the pathogenesis of diabetic nephropathy. Podocytes are characterized by large numbers of mitochondria. However, mitochondrial dysfunction as it relates to kidney pathology remains poorly understood. The present study found that podocyte mitochondria in different animal models of diabetes mellitus became elongated with the development of albuminuria, suggesting a change in mitochondrial dynamics. We then treated cells with a combination of glucose, fatty acids, and angiotensin II (GFA) to mimic the diabetic milieu. Cultured podocytes exposed to GFA showed megamitochondria formation and decreased autophagosome degradation. We also found that GFA treatment decreased the binding of the autophagosome to the lysosome. Our results suggest that megamitochondria are common in podocytes during diabetic nephropathy and that insufficient autophagy flux may underlie this effect. These findings have expanded our understanding of the pathogenesis of diabetic nephropathy and identified a potential pharmacological target for treatment.

Impact of Diabetes Control on Subclinical Atherosclerosis: Analysis from Coronary Computed Tomographic Angiography Registry.

BACKGROUND: There are limited data on the impact of diabetes control on the risk of subclinical coronary atherosclerosis. METHODS: We analyzed 6,434 consecutive asymptomatic individuals without previous history of coronary artery disease who underwent coronary computed tomographic angiography (CCTA) (mean age, 53.7±7.6 years and 4,694 men [73.0%]). The degree and extent of subclinical coronary atherosclerosis were assessed by CCTA, and ≥50% diameter stenosis was defined as significant. A cardiac event was defined as a composite of all-cause death, myocardial infarction, unstable angina, or coronary revascularization. Study participants were categorized as normal (n=5,319), controlled diabetes (glycosylated hemoglobin [HbA1c] <7%, n=747), or uncontrolled diabetes (HbA1c ≥7%, n=368), respectively. RESULTS: Compared with normal individuals, there were no statistically significant differences in the risk of for any atherosclerotic plaque (odds ratio [OR], 1.16; 95% confidence interval [CI], 0.98 to 1.38; P=0.086) and significant coronary artery stenosis (OR, 1.08; 95% CI, 0.82 to 1.42; P=0.583) in controlled diabetic individuals. In contrast, uncontrolled diabetic individuals had consistently higher risks of any atherosclerotic plaque (OR, 2.16; 95% CI, 1.70 to 2.75; P<0.001) and significant coronary artery stenosis (OR, 3.34; 95% CI, 2.52 to 4.43; P<0.001) than normal individuals. During a follow-up of median 5.4 years, there was no significant difference in cardiac events between normal and controlled diabetic individuals (P=0.365). However, uncontrolled diabetes was associated with an increased risk of cardiac events compared with normal individuals (P<0.001) and controlled diabetic individuals (P=0.023). CONCLUSION: Asymptomatic uncontrolled diabetes was associated with significant subclinical coronary atherosclerosis with subsequent high risk for cardiac events.

Inhibition of Ceramide Accumulation in Podocytes by Myriocin Prevents Diabetic Nephropathy.

BACKGROUND: Ceramides are associated with metabolic complications including diabetic nephropathy in patients with diabetes. Recent studies have reported that podocytes play a pivotal role in the progression of diabetic nephropathy. Also, mitochondrial dysfunction is known to be an early event in podocyte injury. Thus, we tested the hypothesis that ceramide accumulation in podocytes induces mitochondrial damage through reactive oxygen species (ROS) production in patients with diabetic nephropathy. METHODS: We used Otsuka Long Evans Tokushima Fatty (OLETF) rats and high-fat diet (HFD)-fed mice. We fed the animals either a control- or a myriocin-containing diet to evaluate the effects of the ceramide. Also, we assessed the effects of ceramide on intracellular ROS generation and on podocyte autophagy in cultured podocytes. RESULTS: OLETF rats and HFD-fed mice showed albuminuria, histologic features of diabetic nephropathy, and podocyte injury, whereas myriocin treatment effectively treated these abnormalities. Cultured podocytes exposed to agents predicted to be risk factors (high glucose, high free fatty acid, and angiotensin II in combination [GFA]) showed an increase in ceramide accumulation and ROS generation in podocyte mitochondria. Pretreatment with myriocin reversed GFA-induced mitochondrial ROS generation and prevented cell death. Myriocin-pretreated cells were protected from GFA-induced disruption of mitochondrial integrity. CONCLUSION: We showed that mitochondrial ceramide accumulation may result in podocyte damage through ROS production. Therefore, this signaling pathway could become a pharmacological target to abate the development of diabetic kidney disease.

Impaired Peroxisomal Fitness in Obese Mice, a Vicious Cycle Exacerbating Adipocyte Dysfunction via Oxidative Stress.

Aims: Peroxisome is a critical organelle for fatty acid oxidation (FAO) and metabolism of reactive oxygen species (ROS). Increased oxidative stress in adipose tissue contributes to the development of insulin resistance and metabolic syndrome in obesity. This study aimed to investigate the role of peroxisomal fitness in maintaining adipocyte function, which has been under-rated in the obesity research area. Results: Reduced peroxisomal gene expressions in white adipose tissue (WAT) of obese mice suggested a close correlation between peroxisomes and obesity. Peroxisomal biogenesis factor 5 siRNA increased cellular ROS and inflammatory mediators in 3T3-L1 adipocytes. On the contrary, hydrogen peroxide or tumor necrosis factor-α treatment significantly decreased biogenesis- and function-related peroxisomal proteins, suggesting a positive feedback loop of ROS/inflammation and peroxisomal dysfunction. Correspondingly, catalase (a major peroxisomal antioxidant)-knockout mice fed with high-fat diet (HFD) exhibited suppressed peroxisomal proteins along with increased oxidative stress and accelerated obesity. In response to fenofibrate (a peroxisomal proliferator) treatment, WAT of HFD-fed wild-type mice showed not only increases in peroxisomal biogenesis and FAO but also attenuated features of adipocyte dysfunction and obesity. However, these results were not observed in peroxisome proliferator-activated receptor-alpha null obese mice. Innovation: Impaired peroxisomal fitness enhanced oxidative stress and inflammation in adipocytes, which exacerbates obesity. Conclusion: Adipose tissue peroxisomal homeostasis plays an important role in attenuating the features of obesity, and it can be a potential therapeutic target of obesity.

Mesenchymal stem cells prevent the progression of diabetic nephropathy by improving mitochondrial function in tubular epithelial cells.

The administration of mesenchymal stem cells (MSCs) was shown to attenuate overt as well as early diabetic nephropathy in rodents, but the underlying mechanism of this beneficial effect is largely unknown. Inflammation and mitochondrial dysfunction are major pathogenic factors in diabetic nephropathy. In this study, we found that the repeated administration of MSCs prevents albuminuria and injury to tubular epithelial cells (TECs), an important element in the progression of diabetic nephropathy, by improving mitochondrial function. The expression of M1 macrophage markers was significantly increased in diabetic kidneys compared with that in control kidneys. Interestingly, the expression of arginase-1 (Arg1), an important M2 macrophage marker, was reduced in diabetic kidneys and increased by MSC treatment. In cultured TECs, conditioned media from lipopolysaccharide-activated macrophages reduced peroxisomal proliferator-activated receptor gamma coactivator 1α (Pgc1a) expression and impaired mitochondrial function. The coculture of macrophages with MSCs increased and decreased the expression of Arg1 and M1 markers, respectively. Treatment with conditioned media from cocultured macrophages prevented activated macrophage-induced mitochondrial dysfunction in TECs. In the absence of MSC coculture, Arg1 overexpression in macrophages reversed Pgc1a suppression in TECs. These observations suggest that MSCs prevent the progression of diabetic nephropathy by reversing mitochondrial dysfunction in TECs via the induction of Arg1 in macrophages.

Nationwide groundwater monitoring around infectious-disease-caused livestock mortality burials in Korea: Superimposed influence of animal leachate on pre-existing anthropogenic pollution.

A foot-and-mouth disease (FMD) outbreak during 2010 affected the entire country of South Korea and approximately 3.4 million swine and bovine mortalities were disposed of at approximately 4800 on-farm burial sites for a few months following the first outbreak. Furthermore, outbreaks of avian influenza (AI) have struck Korea consistently since 2014. Public concern regarding the deterioration of the surrounding environment has been raised aiming at the enormous infected animal carcass burials. On behalf of the Ministry of Environment (ME) of the Korean government, we conducted groundwater monitoring at approximately 3000 wells around the burial sites for 7 years from 2011 to 2017. The baseline groundwater already had enriched levels of nitrogen and chloride compounds masking the influence of leachate with the pre-existing anthropogenic contamination. Based on the trend analysis, most monitoring wells had no significant trends in NO3-N and Cl implying that an abrupt degradation in water quality was not expected across the country. Increasing proportions of total coliform detection in the groundwater of the majority of provinces will cause potential damage to human health around the carcass burial sites. Wells showing frequent upward and downward trends near the carcass burial pits were targeted for principal component analysis (PCA) and the results showed that NH4-N, TOC, Cl, and K could be indicators distinguishing the sole impacts of animal leachate on the groundwater. Analyses of the nationwide monitoring data indicated possibly delayed and prolonged impacts of the widespread burials of livestock mortalities on the groundwater environment. The finding provides realistic insight regarding how to manage the mass burial of livestock mortalities to protect groundwater resources.