Frequency of Pro475Ser Polymorphism of ADAMTS13 Gene and Its Association with ADAMTS-13 Activity in the Korean Population

Purpose: The in vitro study suggested that proline to serine polymorphism in codon 475 (C1423T) of the A Disintegrin and Metalloprotease with ThromboSpondin type 1 repeats-13 (ADAMTS-13) gene is related to reduced activity of ADAMTS-13. In this study, the frequency of the Pro475Ser polymorphism in Koreans was studied and plasma ADAMTS-13 activity was measured to find out whether this polymorphism contributes to decreased ADAMTS-13 activity in Koreans. Patients and Methods: The frequency of the C1423T allele of the ADAMTS13 gene was studied along with measuring plasma ADAMTS-13 activity in 250 healthy Korean individuals. Results: The allele frequency of C1423T polymorphism was 4%, and the median activity of CT type was 107 (69- 143)%, which was lower than in controls with the CC genotype [118 (48- 197)%, (p = 0.021)]. Conclusion: Therefore, the Pro475Ser polymorphism seems to be popular in the Korean population, and attenuates ADAMTS-13 plasma activity. Key Words: ADAMTS-13, polymorphism, thrombotic thromobocytopenic purpur

Effect of Microbial Short-Chain Fatty Acids on CYP3A4-Mediated Metabolic Activation of Human Pluripotent Stem Cell-Derived Liver Organoids

The early and accurate prediction of the hepatotoxicity of new drug targets during nonclinical drug development is important to avoid postmarketing drug withdrawals and late-stage failures. We previously established long-term expandable and functional human-induced pluripotent stem cell (iPSC)-derived liver organoids as an alternative source for primary human hepatocytes. However, PSC-derived organoids are known to present immature fetal characteristics. Here, we treated these liver organoids with microbial short-chain fatty acids (SCFAs) to improve metabolic maturation based on microenvironmental changes in the liver during postnatal development. The effects of the three main SCFA components (acetate, propionate, and butyrate) and their mixture on liver organoids were determined. Propionate (1 µM) significantly promoted the CYP3A4/CYP3A7 expression ratio, and acetate (1 µM), propionate (1 µM), and butyrate (1 µM) combination treatment, compared to no treatment (control), substantially increased CYP3A4 activity and albumin secretion, as well as gene expression. More importantly, mixed SCFA treatment accurately revealed troglitazone-induced hepatotoxicity, which was redeemed on a potent CYP3A4 inhibitor ketoconazole treatment. Overall, we determined, for the first time, that SCFA mixture treatment might contribute to the accurate evaluation of the CYP3A4-dependent drug toxicity by improving metabolic activation, including CYP3A4 expression, of liver organoids

Advanced human liver models for the assessment of drug-induced liver injury

Drug safety issues continue to occur even with drugs that are approved after the completion of clinical studies. Drug-induced liver injury (DILI) is a major obstacle to drug development, because the liver is the primary site of drug metabolism, and injuries caused during this process are severe. Conventional in vitro human liver models, such as 2-dimensional hepatic cell lines, lack in vivo physiological relevance, and animal studies have limitations in the form of species differences and regulatory restrictions. To resolve this issue, an increasing number of 3-dimensional human liver systems, including organoids, are being developed. In this review, we provide an overview of recent assessments of DILI prediction, approaches for in vitro hepatotoxicity evaluation, and a variety of advanced human liver models. We discuss the advantages, limitations, and future perspectives of current human liver models for accurate drug safety evaluations

CFD modelling of cyclonic-DAF (dissolved air flotation) reactor for algae removal

The purpose of this study is to simulate the Cyclonic-DAF tanks using CFD techniques for the optimal design. CFD simulations were performed on two different Cyclonic-DAF reactor shapes either having or not having a cone shaped settled sludge removal unit in the bottom area of the inlet in the Cyclonic-DAF reaction tank. An average flow of less than 10% within the Cyclonic-DAF was rarely found in both shapes while areas of less than 20% were only slightly found near the central concentrate waste pipe. Areas having an average flow of less than 50% were found around the concentrate waste pipe and were observed at a rate of less than 5% of the total volume. Efficiency test results of different Cyclonic-DAF (CDAF) tank height to diameter ratios set at 1:1, 1:35:1, 1.6:1, and 1.85:1 indicated similar efficiencies with turbidity at 91.9% and chlorophyll-a at 94.7%. Keywords: Algae removal, Dissolved air flotation, DAF, Cyclonic-DAF, Computational fluid dynamics (CFD

Recent advances in multicellular human liver models

The liver is the most important metabolic organ in the body. Model systems that recapitulate the complex organ structure and cell composition of the human liver are insufficient to study liver biology and to test toxicity and efficacy during new drug development. Recently established 3-dimensional liver models, including spheroids and organoids, organs-on-a-chip, bioprinting, and the decellularization/recellularization technique, have provided platforms that emulate the structural and functional characteristics of the human liver better than conventional 2-dimensional cell culture models and animal models. This review summarizes the architecture and cell compositions of human liver tissue, focusing on recent studies of multicellular human liver models that recapitulate in vivo-like physiologies with morphological and functional advances by the cellular communication of parenchymal and non-parenchymal cells. We discuss the applications, limitations, and future perspectives of advanced multicellular human liver models

Standard operating protocol of hepatic organoid differentiation from human induced pluripotent stem cells

Background Mature liver organoids are potential cellular sources for research to understand the pathology of several liver-related conditions, including end-stage chronic liver disease. Although several methods exist for the differentiation of mature hepatic organoids from human induced pluripotent stem cells (hiPSCs), organoid generation have limitations due to various experimental culture conditions. Methods We differentiated homozygous hiPSCs into definitive endoderm, hepatic endoderm, and hepatic maturation in a two-dimensional culture condition. Expandable hepatic organoids (EHO) and mature hepatic organoids (MHO) were cultured in a three-dimensional culture condition. Results We successfully established hepatic organoids that were reproductively and expandably cultured for at least three months. MHO showed significant increases in hepatic-specific markers, drug-metabolizing enzymes and transporter genes, and human albumin secretion. Conclusion Therefore, we established a standard operating protocol for gen¬erating mature and expandable hepatic organoids derived from hiPSCs, and made the start¬ing materials available to promote widespread use of the protocol

Efficient and reproducible generation of human induced pluripotent stem cell-derived expandable liver organoids for disease modeling

Abstract Genetic liver disease modeling is difficult because it is challenging to access patient tissue samples and to develop practical and relevant model systems. Previously, we developed novel proliferative and functional liver organoids from pluripotent stem cells; however, the protocol requires improvement for standardization and reproducible mass production. Here, we improved the method such that it is suitable for scalable expansion and relatively homogenous production, resulting in an efficient and reproducible process. Moreover, three medium components critical for long-term expansion were defined. Detailed transcriptome analysis revealed that fibroblast growth factor signaling, the essential pathway for hepatocyte proliferation during liver regeneration, was mainly enriched in proliferative liver organoids. Short hairpin RNA-mediated knockdown of FGFR4 impaired the generation and proliferation of organoids. Finally, glycogen storage disease type Ia (GSD1a) patient-specific liver organoids were efficiently and reproducibly generated using the new protocol. They well maintained disease-specific phenotypes such as higher lipid and glycogen accumulation in the liver organoids and lactate secretion into the medium consistent with the main pathologic characteristics of patients with GSD1a. Therefore, our newly established liver organoid platform can provide scalable and practical personalized disease models and help to find new therapies for incurable liver diseases including genetic liver diseases

The prevalence and clinical significance of transitional vertebrae: a radiologic investigation using whole spine spiral three-dimensional computed tomographic images

Background Errors in counting spinal segments are common during interventional procedures when there are transitional vertebrae. In this study, we investigated the prevalence of the transitional vertebrae including thoracolumbar transitional vertebra (TLTV) and lumbosacral transitional vertebrae (LSTV). The relationship between the existence of TLTV and abnormal rib count or the existence of LSTV were also evaluated. Methods The vertebral levels were counted craniocaudally, starting from C1, based on the assumption of 7 cervical, 12 thoracic, and 5 lumbar vertebrae, using whole spine spiral three-dimensional computed tomographic images. The 20th and 25th vertebrae were defined as L1 and S1, respectively. Results In total, 150 patients had TLTV, with a prevalence of 11.2% (150/1,340). LSTV was observed in 111 of 1,340 cases (8.3%). Sacralization was observed in 68 of 1,340 cases (5.1%) and lumbarization in 43 of 1,340 cases (3.2%). There was a significant relationship between the existence of TLTV and the abnormal rib count (odds ratio [OR]: 117.26, 95% confidence interval [95% CI]: 60.77–226.27; P < 0.001) and LSTV (OR: 7.38, 95% CI: 3.99–13.63; P < 0.001). Conclusions Our study results suggest that patients with TLTV are more likely to have an abnormal rib count or LSTV. If a TLTV or LSTV is seen on the fluoroscopic image, a whole spine image is necessary to permit accurate numbering of the lumbar vertebra

Genomic Signature of the Standardized Uptake Value in 18F-Fluorodeoxyglucose Positron Emission Tomography in Breast Cancer

The standardized uptake value (SUV), an indicator of the degree of glucose uptake in 18F-fluorodeoxyglucose positron emission tomography (FDG-PET), has been used for predicting the clinical behavior of malignant tumors. However, its characteristics have been insufficiently explored at the genomics level. Here, we aim to identify genomic signatures reflecting prognostic SUV characteristics in breast cancer (BRC). Through integrative genomic profiling of 3710 BRC patients, including 254 patients who underwent preoperative FDG-PET, we identified an SUV signature, which showed independent clinical utility for predicting BRC prognosis (hazard ratio [HR] 1.27, 95% confidence interval [CI] = 1.12 to 1.45, p = 2.23 &times; 10&minus;4). The risk subgroups classified by the signature exhibited mutually exclusive mutation patterns of TP53 and PIK3CA and showed significantly different responsiveness to immunotherapy. Experimental assays revealed that a signaling axis defined by TP53&ndash;FOXM1 and its downstream effectors in glycolysis&ndash;gluconeogenesis, including LDHA, might be important mediators in the FDG-PET process. Our molecular characterizations support an understanding of glucose metabolism and poor prognosis in BRC with a high SUV, utilizable in clinical practice to assist other diagnostic tools