Biocompatible and Biodegradable Fe³⁺–Melanoidin Chelate as a Potentially Safe Contrast Agent for Liver MRI

Currently, most MRI probes available for clinical use contain gadolinium, which is a high-risk paramagnetic metal that can cause severe side effects (e.g., nephrogenic systemic fibrosis). To limit such side effects and improve diagnostic efficacy, we developed a novel biocompatible MRI contrast agent using glucose, glycine, and paramagnetic iron ion. Glucose and glycine were polymerized into melanoidin by the nonenzymatic Maillard reaction, and Fe3+ was chelated stably with the melanoidin during polymerization. The Fe3+–melanoidin chelate had biocompatibility, biodegradability, and unique contrast effects on both T1- and T2-weighted MRI, depending on the pH and oxidative environments. The administration of the Fe3+–melanoidin chelate to a mouse model of liver cancer showed highly enhanced liver-to-tumor contrasts on both T1- and T2-weighted MRI

Data_Sheet_1_Monthly Variation in Flux of Inorganic Nutrients From Submarine Groundwater Discharge in a Volcanic Island: Significant Nitrogen Contamination in Groundwater.docx

To determine nutrient fluxes derived from submarine groundwater discharge (SGD), we conducted monthly hydrological surveys on the coast of Jeju, a volcanic island located in the southern sea of Korea. The concentrations of dissolved inorganic nitrogen (DIN), dissolved inorganic phosphorus (DIP), and dissolved silicon (DSi) were significantly correlated with salinity, indicating that fresh SGD (FSGD) is a major nutrient source in Jeju Island where no other coastal freshwater origins exist. Based on a DSi-mass balance model, seepage rate of FSGD was found to depend on 5-day precipitation before sampling campaigns, which immediately permeated via porous aquifers. Thus, the FSGD-driven nutrient fluxes were generally higher in rainy season (July–August) and September 2019 when typhoons occurred. However, high DIN and DIP fluxes were found during spring (March–May), even at low seepage rate, perhaps by a fertilizer input from agriculture activity. This study highlights that large variation of the SGD-driven nutrient fluxes was caused by environmental and anthropogenic factors and emphasizes on the importance of long-term investigation.</p

Target-Specific Gene Silencing of Layer-by-Layer Assembled Gold–Cysteamine/siRNA/PEI/HA Nanocomplex

Target-specific intracellular delivery of small interfering RNA (siRNA) is regarded as one of the most important technologies for the development of siRNA therapeutics. In this work, a cysteamine modified gold nanoparticles (AuCM)/siRNA/polyethyleneimine (PEI)/hyaluronic acid (HA) complex was successfully developed using a layer-by-layer method for target-specific intracellular delivery of siRNA by HA receptor mediated endocytosis. Atomic force microscopic and zeta potential analyses confirmed the formation of a AuCM/siRNA/PEI/HA complex having a particle size of ca. 37.3 nm and a negative surface charge of ca. −12 mV. With a negligible cytotoxicity, AuCM/siRNA/PEI/HA complex showed an excellent target-specific gene silencing efficiency of ca. 70% in the presence of 50 vol % serum, which was statistically much higher than that of siRNA/Lipofectamine 2000 complex. In the competitive binding tests with free HA, dark-field bioimaging and inductively coupled plasma–atomic emission spectroscopy confirmed the target-specific intracellular delivery of AuCM/siRNA/PEI/HA complex to B16F1 cells with HA receptors. Moreover, the systemic delivery of AuCM/siRNA/PEI/HA complex using apolipoprotein B (ApoB) siRNA as a model drug resulted in a significantly reduced ApoB mRNA level in the liver tissue. Taken together, AuCM/siRNA/PEI/HA complex was thought to be developed as target-specific siRNA therapeutics for the systemic treatment of various liver diseases

KLF8 directly binds to C/EBPα and PPARγ2 promoter.

<p>(A) EMSA experiment using KLF binding sites of β-globin (positive control), C/EBPα, or PPARγ2 promoter as probes. A FLAG-tagged KLF8 protein was in vitro translated using TNT T7 quick master mix. (B) EMSA experiment using wild type or mutated probe of the C/EBPα promoter. FLAG-tagged C/EBPβ-LAP or KLF8 protein was in vitro translated using TNT T7 quick master mix. (C) Western blot analysis of the in vitro translated proteins. n.s, non-specific. (D) ChIP was performed on 3T3-L1 cell chromatin at the indicated time points after induction of differentiation, using control IgG, anti-C/EBPβ, or anti-KLF8 antibody. The immunoprecipitated DNA was used as a PCR template to detect the PPARγ or C/EBPα promoter regions.</p

Overexpression of KLF8 results in enhanced differentiation.

<p>Preadipocyte 3T3-L1 cells were transiently transfected using electroporation with pcDNA3.0-KLF8-FLAG or empty vector and differentiated with standard hormonal cocktail as described in Materials and Methods. (A) After 5 days of differentiation, lipid accumulation was detected by oil red-O staining. (B) Western blot analysis of whole-cell extracts prepared at the indicated times during differentiation. LAP, liver-activating protein, LIP, liver-inhibitory protein, PPARγ1, PPARγ2, and 43 or 30 kDa of C/EBPα proteins were indicated.</p

Univariate and multivariate analysis for overall survival.

<p>Univariate and multivariate analysis for overall survival.</p

Trends of participating in clinical trials among patients with MBC from 2000 to 2014.

<p>Trends of participating in clinical trials among patients with MBC from 2000 to 2014.</p

KLF8 expression during 3T3-L1 adipocyte differentiation and in mouse adipose tissue.

<p>(A) Total RNA was extracted from 3T3-L1 cells at the indicated times before and after induction of differentiation. The expression levels of KLF5, KLF8, KLF12, and KLF17 were determined by RT-PCR. (B) Anti-rabbit polyclonal KLF8 antibody was generated and tested on 293T cells that were transfected with either pcDNA3.0 or pcDNA3.0-KLF8-FLAG. Whole-cell lysates were immunoblotted (IB) using anti-KLF8 or anti-FLAG, which verified the specific antigen-antibody interaction. (C) Western blot analysis of KLF8 expression was performed at the indicated time points during 3T3-L1 cell differentiation. (D) KLF5, KLF8, and fatty acid synthase (FASN) mRNA levels were measured in the stromal vascular fraction (SVF) or the fat fraction of mouse epididymal adipose tissue using real-time qPCR. Data represent the mean ± SD.</p

Real-world treatment persistence of non-tumor necrosis factor inhibitors versus tumor necrosis factor inhibitors among patients with rheumatoid arthritis in South Korea

Aims: We aimed to assess treatment persistence of tumor necrosis factor (TNF) inhibitors and non-TNF inhibitors in two groups of rheumatoid arthritis (RA) patients: biologic disease-modifying antirheumatic drug (bDMARD) initiators and switchers. Patients and methods: This retrospective cohort study utilized a national health insurance claims database. Patients aged ≥18 years initiating/switching bDMARD between 1 December 2013 and 31 December 2014, the index period, were followed for 12 months. Initiators who began treatment with a bDMARD during the index period were defined as having no bDMARD prescriptions for the previous year. Switchers who changed treatment from the previous bDMARD to the index bDMARD were defined as having different bDMARDs during the index period. Treatment persistence rates during the follow-up period were measured, and factors associated with non-persistence were assessed with the Cox proportional hazard model. Results: Of 2684 patients, treatment persistence rates were the highest for abatacept in initiators (69.3%) and tocilizumab in switchers (77.0%), while adalimumab showed the lowest persistence rates for both initiators and switchers (48.2%, 28.8%), followed by etanercept (51.3%, 41.0%). Adalimumab and etanercept were significantly more likely to show non-persistence (HR 1.58, 95% CI 1.27–1.96; HR 1.42, 95% CI 1.14–1.76) compared to infliximab for initiators, while tocilizumab was significantly more likely to show persistence (HR 0.411, 95% CI 0.206–0.819) in switchers. Conclusions: Non-TNF inhibitors showed higher persistence rates than TNF inhibitors in South Korean RA patients, and tocilizumab especially was associated with higher persistence in patients with inadequate response to TNF inhibitors. Good persistence with non-TNF inhibitors indicates the potential for long-term efficacy as first-line treatment.</p

Gene expression change of KLF family members during 3T3-L1 differentiation.

*<p>Microarray was performed using total RNA samples from 3T3-L1 cells after 0, 2, 4, and 7 days of differentiation. Gene expression was analyzed by Agilent Mouse Genome 4×44 k oligo chip. The preparation for hybridization and the scanning of mouse chips were performed according to the manufacturer’s protocols (Genocheck). More than 2-fold changes of expression are indicated in bold.</p