HLA DR Genome Editing with TALENs in Human iPSCs Produced Immune-Tolerant Dendritic Cells.

Although human induced pluripotent stem cells (iPSCs) can serve as a universal cell source for regenerative medicine, the use of iPSCs in clinical applications is limited by prohibitive costs and prolonged generation time. Moreover, allogeneic iPSC transplantation requires preclusion of mismatches between the donor and recipient human leukocyte antigen (HLA). We, therefore, generated universally compatible immune nonresponsive human iPSCs by gene editing. Transcription activator-like effector nucleases (TALENs) were designed for selective elimination of HLA DR expression. The engineered nucleases completely disrupted the expression of HLA DR on human dermal fibroblast cells (HDF) that did not express HLA DR even after stimulation with IFN-γ. Teratomas formed by HLA DR knockout iPSCs did not express HLA DR, and dendritic cells differentiated from HLA DR knockout iPSCs reduced CD4+ T cell activation. These engineered iPSCs might provide a novel translational approach to treat multiple recipients from a limited number of cell donors.

Anti-inflammatory evaluation of the methanolic extract of Taraxacum officinale in LPS-stimulated human umbilical vein endothelial cells.

BACKGROUND: Atherosclerosis is a chronic vascular inflammatory disease. Since even low-level endotoxemia constitutes a powerful and independent risk factor for the development of atherosclerosis, it is important to find therapies directed against the vascular effects of endotoxin to prevent atherosclerosis. Taraxacum officinale (TO) is used for medicinal purposes because of its choleretic, diuretic, antioxidative, anti-inflammatory, and anti-carcinogenic properties, but its anti-inflammatory effect on endothelial cells has not been established. METHODS: We evaluated the anti-inflammatory activity of TO filtered methanol extracts in LPS-stimulated human umbilical vein endothelial cells (HUVECs) by monocyte adhesion and western blot assays. HUVECs were pretreated with 100 µg/ml TO for 1 h and then incubated with 1 µg/ml LPS for 24 h. The mRNA and protein expression levels of the targets (pro-inflammatory cytokines and adhesion molecules) were analyzed by real-time PCR and western blot assays. We also preformed HPLC analysis to identify the components of the TO methanol extract. RESULTS: The TO filtered methanol extracts dramatically inhibited LPS-induced endothelial cell-monocyte interactions by reducing vascular cell adhesion molecule-1 and monocyte chemoattractant protein-1, and pro-inflammatory cytokine expression. TO suppressed the LPS-induced nuclear translocation of NF-κB, whereas it did not affect MAPK activation. CONCLUSIONS: Our findings demonstrated that methanol extracts of TO could attenuate LPS-induced endothelial cell activation by inhibiting the NF-κB pathway. These results indicate the potential clinical benefits and applications of TO for the prevention of vascular inflammation and atherosclerosis.

Characteristics of Body Composition and Muscle Strength of North Korean Refugees during South Korean Stay.

BACKGROUND: The aim of this study was to investigate the changes of body composition and muscle strength of North Korean refugees (NKRs) according to their duration of stay in South Korea. METHODS: NKRs who volunteered and were living in South Korea, aged 20 to 75 years were recruited. Body compositions were analyzed by bioelectrical impedance analysis. Muscle strength was measured with the hand grip test. Demographic and migration information was obtained with a questionnaire. RESULTS: A total of 158 volunteers were recruited at a mean age of 48.3±11.4 years. The mean time from when they escaped from North Korea and arrived in South Korea was 5.8±4.3 years. Height, weight, and body surface area were significantly smaller in all NKRs compared to South Korean controls, except for women aged over 50 years. In females of younger ages (<50 years), NKRs with more than a 4-year stay in South Korea had a higher weight and fat mass than that of those who had a shorter stay (less than 4 years) in South Korea. All NKRs had a weaker grip strength than that of the age-matched controls from South Korea. CONCLUSION: The NKRs showed relatively smaller physiques and weaker muscle strength than that of the South Korean controls. In younger female NKRs, shorter South Korean stay group showed small body weight and fat mass than that of longer South Korean stay group. Specific health support programs might be needed.

Fundamental issues related to the origin of melatonin and melatonin isomers during evolution: relation to their biological functions.

Melatonin and melatonin isomers exist and/or coexist in living organisms including yeasts, bacteria and plants. The levels of melatonin isomers are significantly higher than that of melatonin in some plants and in several fermented products such as in wine and bread. Currently, there are no reports documenting the presence of melatonin isomers in vertebrates. From an evolutionary point of view, it is unlikely that melatonin isomers do not exist in vertebrates. On the other hand, large quantities of the microbial flora exist in the gut of the vertebrates. These microorganisms frequently exchange materials with the host. Melatonin isomers, which are produced by these organisms inevitably enter the host's system. The origins of melatonin and its isomers can be traced back to photosynthetic bacteria and other primitive unicellular organisms. Since some of these bacteria are believed to be the precursors of mitochondria and chloroplasts these cellular organelles may be the primary sites of melatonin production in animals or in plants, respectively. Phylogenic analysis based on its rate-limiting synthetic enzyme, serotonin N-acetyltransferase (SNAT), indicates its multiple origins during evolution. Therefore, it is likely that melatonin and its isomer are also present in the domain of archaea, which perhaps require these molecules to protect them against hostile environments including extremely high or low temperature. Evidence indicates that the initial and primary function of melatonin and its isomers was to serve as the first-line of defence against oxidative stress and all other functions were acquired during evolution either by the process of adoption or by the extension of its antioxidative capacity.

Delivery of pineal melatonin to the brain and SCN: role of canaliculi, cerebrospinal fluid, tanycytes and Virchow-Robin perivascular spaces.

Historically, the direct release of pineal melatonin into the capillary bed within the gland has been accepted as the primary route of secretion. Herein, we propose that the major route of melatonin delivery to the brain is after its direct release into the cerebrospinal fluid (CSF) of the third ventricle (3V). Melatonin concentrations in the CSF are not only much higher than in the blood, also, there is a rapid nocturnal rise at darkness onset and precipitous decline of melatonin levels at the time of lights on. Because melatonin is a potent free radical scavenger and antioxidant, we surmise that the elevated CSF levels are necessary to combat the massive free radical damage that the brain would normally endure because of its high utilization of oxygen, the parent molecule of many toxic oxygen metabolites, i.e., free radicals. Additionally, the precise rhythm of CSF melatonin provides the master circadian clock, the suprachiasmatic nucleus, with highly accurate chronobiotic information regarding the duration of the dark period. We predict that the discharge of melatonin directly into the 3V is aided by a number of epithalamic structures that have heretofore been overlooked; these include interpinealocyte canaliculi and evaginations of the posterodorsal 3V that directly abut the pineal. Moreover, the presence of tanycytes in the pineal recess and/or a discontinuous ependymal lining in the pineal recess allows melatonin ready access to the CSF. From the ventricles melatonin enters the brain by diffusion and by transport through tanycytes. Melatonin-rich CSF also circulates through the aqueduct and eventually into the subarachnoid space. From the subarachnoid space surrounding the brain, melatonin penetrates into the deepest portions of the neural tissue via the Virchow-Robin perivascular spaces from where it diffuses into the neural parenchyma. Because of the high level of pineal-derived melatonin in the CSF, all portions of the brain are better shielded from oxidative stress resulting from toxic oxygen derivatives.

The prevalence of and factors associated with the use of smoking cessation medication in Korea: trend between 2005-2011.

BACKGROUND: In Korea, nicotine replacement therapy (NRT) has been widely used in government-led, public health center-based smoking cessation services since 2004 and varenicline has become available from 2007 but without reimbursement. In this study which used a series of nationwide cross-sectional surveys in Korea performed from 2005 to 2011, we examined the prevalence of smoking cessation medication use and factors associated with it. METHODS: We analyzed data from the third to fifth waves of Korean National Health and Nutrition Examination Survey (2005-2011). Prevalence of each smoking cessation method use was calculated for each year, and its secular trend was tested by multivariate logistic regression. RESULTS: Among smokers who made quit attempt during the previous year, 15.7% had used smoking cessation medications,15.3% had used NRT, and 0.7% had used prescription medication. There was a significant increasing trend for NRT use (P<0.001) during the study period, but use of prescription medication did not show any increase over time (P = 0.654) Education on smoking prevention and cessation was associated with smoking cessation medications use (OR 2.08, 95% CI 1.58-2.75). CONCLUSIONS: While the use of NRT has increased over years through government-sponsored smoking cessation programs, use of prescription drugs remained very low and flat probably due to lack of reimbursement. Education of smokers about effective smoking cessation methods and change in reimbursement policy are suggested to stimulate evidence-based smoking cessation practice.

Metabolic effects of aloe vera gel complex in obese prediabetes and early non-treated diabetic patients: randomized controlled trial.

OBJECTIVE: The metabolic effects of an aloe vera gel complex (Aloe QDM complex) on people with prediabetes or early diabetes mellitus (DM) are unknown. The goal of this study was to determine the effects of Aloe QDM complex on body weight, body fat mass (BFM), fasting blood glucose (FBG), fasting serum insulin, and Homeostasis Model of Assessment - Insulin Resistance (HOMA-IR) in obese individuals with prediabetes or early DM who were not on diabetes medications. METHODS: Participants (n = 136) were randomly assigned to an intervention or a control group and evaluated at baseline and at 4 and 8 wk. RESULTS: The study lost six participants in the control group and eight in the intervention group. At 8 wk, body weight (P = 0.02) and BFM (P = 0.03) were significantly lower in the intervention group. At 4 wk, serum insulin level (P = 0.04) and HOMA-IR (P = 0.047) were lower in the intervention group; they also were lower at 8 wk but with borderline significance (P = 0.09; P = 0.08, respectively). At 8 wk, FBG tended to decrease in the intervention group (P = 0.02), but the between-group difference was not significant (P = 0.16). CONCLUSION: In obese individuals with prediabetes or early untreated DM, Aloe QDM complex reduced body weight, BFM, and insulin resistance.

Prevalence, awareness, and treatment of osteoporosis among Korean women: The Fourth Korea National Health and Nutrition Examination Survey.

PURPOSE: This study aimed to assess the factors associated with the prevalence, awareness, and treatment of osteoporosis in a representative sample of Korean women. METHODS: Data were obtained from dual energy X-ray absorptiometry measurement of the lumbar vertebrae and femoral neck, and from a standardized questionnaire in 2870 Korean women aged 50years and older who participated in the Fourth Korea National Health and Nutrition Examination Survey 2008-2009. Osteoporosis was defined by World Health Organization T-score criteria, and awareness and treatment were defined by self-report of an osteoporosis diagnosis and self-report of current anti-osteoporotic medication use, respectively. We assessed the relationship between multiple risk factors and prevalence, awareness, and treatment. RESULTS: Osteoporosis was reported in 39.1% of Korean women. Among those with osteoporosis, only 37.5% were aware of their diagnosis and 23.5% received pharmacological treatment. Despite higher prevalence among respondents who were older, of lower body weight, calcium intake, physical activity, and education levels, the awareness and treatment rates of these groups were similar or lower than that of the low-risk controls in multivariate logistic regression models. Moreover, easily identifiable risk factors (e.g., history of fracture, falls, height loss, familial osteoporosis) were not associated with awareness and treatment. Participants who had undergone health screening in the previous 2years exhibited increased awareness and treatment rates independently of other demographic factors. CONCLUSIONS: Osteoporosis was highly prevalent in this Korean study but was underdiagnosed and undertreated. Routine health screenings could be an effective strategy to increase osteoporosis awareness and treatment.

Solubilization of water soluble anthocyanins in apolar medium using reverse micelle.

Crude anthocyanins extracted from grape skin were solubilized in hexane containing 100 mM bis(2-ethylhexyl)sodium sulfosuccinate (AOT) by forming stable reverse micelles (RMs). Anthocyanins solubilized in RMs showed about four times greater color intensity than that in aqueous medium. The color intensity of anthocyanins in RMs was primarily affected by the interaction between sulfonate head of AOT and flavylium cation of anthocyanins. The molar ratio of water to AOT (Wo) also influenced the color properties. As the Wo increased from five to 20, the color intensity increased and resulted in a bathochromic shift. This result suggests that increased micelle size facilitates complexation between AOT and flavylium cation. The color stability of anthocyanins in RM was higher than that of buffered anthocyanins during the storage at 30 degrees C. The current study might be utilized as a model system to predict color properties of anthocyanins in apolar medium.

Melatonin prevents oxidative damage to protein and lipid induced by ascorbate-Fe(3+)-EDTA: Comparison with glutathione and alpha-tocopherol.

OBJECTIVES: The ability of melatonin to protect protein and lipid against oxidative damage induced by an ascorbate-Fe(3+)-EDTA (AFE) system which generates the hydroxyl radical was investigated using bovine serum albumin (BSA) and phosphatidylcholine (PC) liposomes, respectively, and compared with the protective effects of reduced glutathione and alpha-tocopherol. The comparison study was also performed using PC liposomes containing BSA. METHODS: BSA, PC liposomes or their mixtures were exposed to the HO.-generating system of AFE composed of 0.1 mM EDTA-Fe(3+) and 0.5 mM ascorbate in 0.1 M phosphate buffer, pH 7.4, at 37 degrees C for 1 h. Oxidative damage of BSA was determined by measuring the carbonyl content and the fragmentation of protein by the reaction with dinitrophenylhydrazine (DNPH) and electrophoresis, respectively. Lipid peroxidation of PC liposomes was indicated by the quantity of malondialdehyde and 4hydroxyalkenals. RESULTS: Melatonin inhibited protein damage as indicated by the reduced formation of carbonyl groups and fragmentation of BSA by AFE as effectively as did glutathione while alpha-tocopherol was ineffective. Melatonin also prevented lipid peroxidation to the same extent as did alpha-tocopherol in PC liposomes. CONCLUSION: Both BSA and PC lipid exposed to AFE are effectively protected by melatonin while hydrophilic glutathione and hydrophobic alpha-tocopherol are as effective as melatonin only in one target, i.e., BSA or PC lipid, respectively.