Neutral sphingomyelinase blockade enhances hematopoietic stem cell fitness through an integrated stress response.

Hematopoietic stem and progenitor cell (HSPC) transplantation serves as a curative therapy for many benign and malignant hematopoietic disorders and as a platform for gene therapy. However, growing needs for ex vivo manipulation of HSPC-graft products are limited by barriers in maintaining critical self-renewal and quiescence properties. The role of sphingolipid metabolism in safeguarding these essential cellular properties has been recently recognized, but not yet widely explored. Here, we demonstrate that pharmacologic and genetic inhibition of neutral sphingomyelinase 2 (nSMase-2) leads to sustained improvements in long-term competitive transplantation efficiency after ex vivo culture. Mechanistically, nSMase-2 blockade activates a canonical integrated stress response (ISR) and promotes metabolic quiescence in human and murine HSPCs. These adaptations result in part from disruption in sphingolipid metabolism that impairs the release of nSMase-2-dependent extracellular vesicles (EVs). The aggregate findings link EV trafficking and the ISR as a regulatory dyad guarding HSPC homeostasis and long-term fitness. Translationally, transient nSMase-2 inhibition enables ex vivo graft manipulation with enhanced HSPC potency.

A novel thiazolidinedione derivative TD118 showing selective algicidal effects for red tide control.

Thiazolidinedione (TD) derivatives have been found to have an algicidal effect on harmful algal bloom microalgae. In this study, 75 TD derivatives were synthesized and analyzed for algicidal activity. Among these synthetic TDs, 18 TD derivatives showed specific algicidal activity on two strains belonging to the classes Raphidophyceae (Chattonella marina and Heterosigma akashiwo) and Dinophyceae (Cochlodinium polykrikoides). Two strains belonging to Bacillariophyceae (Navicula pelliculosa and Phaeodactylum EPV), one strain belonging to Dinophyceae (Amphidinium sp.), and a Eustigmatophycean microalga (Nannochloropsis oculata) showed less sensitivity to the TD derivatives than the other two phyla. The most reactive TD derivative, compound 2 (TD118), was selected and tested for morphological and physiological changes. TD118 effectively damaged the cell membrane of C. marina, H. akashiwo and C. polykrikoides. The O2 evolution and photosystem II efficiency (F(v)/F(m)) of C. marina, H. akashiwo and C. polykrikoides were also severely reduced by TD118 treatment. Amphidinium sp., N. pelliculosa, Phaeodactylum EPV and N. oculata showed less reduction of O2 evolution and the F(v)/F(m) by TD118. These results imply that the species-specific TD structure relationship may be due to structural and/or physiological differences among microalgal species.

Dynamic Tracking of Native Polyclonal Hematopoiesis in Adult Mice.

Hematopoietic dysfunction has been associated with a reduction in the number of active precursors. However, precursor quantification at homeostasis and under diseased conditions is constrained by the scarcity of available methods. To address this issue, we optimized a method for quantifying a wide range of hematopoietic precursors. Assuming the random induction of a stable label in precursors following a binomial distribution, the estimation depends on the inverse correlation between precursor numbers and the variance of precursor labeling among independent samples. Experimentally validated to cover the full dynamic range of hematopoietic precursors in mice (1 to 105), we utilized this approach to demonstrate that thousands of precursors, which emerge after modest expansion during fetal-to-adult transition, contribute to native and perturbed hematopoiesis. We further estimated the number of precursors in a mouse model of Fanconi Anemia, showcasing how repopulation deficits can be segregated into autologous (cell proliferation) and non-autologous causes (lack of precursor). Our results support an accessible and reliable approach for precursor quantification, emphasizing the contemporary perspective that native hematopoiesis is highly polyclonal.

Inflammatory recruitment of healthy hematopoietic stem and progenitor cells in the acute myeloid leukemia niche.

Inflammation in the bone marrow (BM) microenvironment is a constitutive component of leukemogenesis in acute myeloid leukemia (AML). Current evidence suggests that both leukemic blasts and stroma secrete proinflammatory factors that actively suppress the function of healthy hematopoietic stem and progenitor cells (HSPCs). HSPCs are also cellular components of the innate immune system, and we reasoned that they may actively propagate the inflammation in the leukemic niche. In two separate congenic models of AML we confirm by evaluation of the BM plasma secretome and HSPC-selective single-cell RNA sequencing (scRNA-Seq) that multipotent progenitors and long-lived stem cells adopt inflammatory gene expression programs, even at low leukemic infiltration of the BM. In particular, we observe interferon gamma (IFN-γ) pathway activation, along with secretion of its chemokine target, CXCL10. We show that AML-derived nanometer-sized extracellular vesicles (EVAML) are sufficient to trigger this inflammatory HSPC response, both in vitro and in vivo. Altogether, our studies indicate that HSPCs are an unrecognized component of the inflammatory adaptation of the BM by leukemic cells. The pro-inflammatory conversion and long-lived presence of HSPCs in the BM along with their regenerative re-expansion during remission may impact clonal selection and disease evolution.

Genome-wide association study revealed the genomic regions associated with skin pigmentation in an Ogye x White Leghorn F2 chicken population.

Skin color in chickens is an economically important trait that determines the first impression of a consumer toward a broiler and can ultimately affect consumer choice in the market. Therefore, identification of genomic regions associated with skin color is crucial for increasing the sales value of chickens. Although previous studies have attempted to reveal the genetic markers associated with the skin coloration in chickens, most were limited to investigations of candidate genes, such as melanin-related genes, and focused on case/control studies based on a single or small population. In this study, we performed a genome-wide association study (GWAS) on 770 F2 intercrosses produced by an experimental population of 2 chicken breeds, namely Ogye and White Leghorns, with different skin colors. The GWAS demonstrated that the L* value among the 3 skin color traits is highly heritable, and the genomic regions located on 2 chromosomes (20 and Z) were detected to harbor SNPs significantly associated with the skin color trait, accounting for most of the total genetic variance. Particular genomic regions spanning a ∼2.94 Mb region on GGA Z and a ∼3.58 Mb region on GGA 20 were significantly associated with skin color traits, and in these regions, certain candidate genes, including MTAP, FEM1C, GNAS, and EDN3, were found. Our findings could help elucidate the genetic mechanisms underlying chicken skin pigmentation. Furthermore, the candidate genes can be used to provide a valuable breeding strategy for the selection of specific chicken breeds with ideal skin coloration.

Rational biomarker development for the early and minimally invasive monitoring of AML.

Recurrent disease remains the principal cause for treatment failure in acute myeloid leukemia (AML) across age groups. Reliable biomarkers of AML relapse risk and disease burden have been problematic, as symptoms appear late and current monitoring relies on invasive and cost-ineffective serial bone marrow (BM) surveillance. In this report, we discover a set of unique microRNA (miRNA) that circulates in AML-derived vesicles in the peripheral blood ahead of the general dissemination of leukemic blasts and symptomatic BM failure. Next-generation sequencing of extracellular vesicle-contained small RNA in 12 AML patients and 12 controls allowed us to identify a panel of differentially incorporated miRNA. Proof-of-concept studies using a murine model and patient-derived xenografts demonstrate the feasibility of developing miR-1246, as a potential minimally invasive AML biomarker.

Hematopoietic stem and progenitor cell signaling in the niche.

Hematopoietic stem and progenitor cells (HSPCs) are responsible for lifelong maintenance of hematopoiesis through self-renewal and differentiation into mature blood cell lineages. Traditional models hold that HSPCs guard homeostatic function and adapt to regenerative demand by integrating cell-autonomous, intrinsic programs with extrinsic cues from the niche. Despite the biologic significance, little is known about the active roles HSPCs partake in reciprocally shaping the function of their microenvironment. Here, we review evidence of signals emerging from HSPCs through secreted autocrine or paracrine factors, including extracellular vesicles, and via direct contact within the niche. We also discuss the functional impact of direct cellular interactions between hematopoietic elements on niche occupancy in the context of leukemic infiltration. The aggregate data support a model whereby HSPCs are active participants in the dynamic adaptation of the stem cell niche unit during development and homeostasis, and under inflammatory stress, malignancy, or transplantation.

Development of a fast real-time PCR assay based on TaqMan probe for identification of edible rice grasshopper (Oxya chinensis) in processed food products.

Interest in using insects as an alternative source of food for humans is increasing. However, few analytical methods provide accurate information about the presence of insect species in processed foods. In this study, we developed a fast real-time PCR assay based on a TaqMan probe that can be performed within 40 min to detect edible rice grasshopper in commercial food products. A rice grasshopper-specific primer pair and probe targeting the cytochrome c oxidase subunit 1 (COI) gene were newly designed, having an amplicon size of 110 bp. The specificity of this primer pair and probe was verified using 19 insects and five crustaceans and no cross-reactivity was obtained against the non-target species. The absolute limit of detection (LOD) was 0.5 pg of rice grasshopper DNA, and as low as 0.1% of rice grasshopper was detected in raw, heat-treated, and autoclaved binary insect mixtures. To evaluate the effect of food matrix, binary mixtures containing rice grasshopper in wheat were used additionally, and at least 0.1% of target species was detected using this assay. The applicability of this assay was confirmed using nine commercial food samples labeled as containing rice grasshopper or locust. The fast real-time PCR developed in this study is a specific and sensitive method for identifying edible rice grasshopper in various food samples.

Neural network control for position tracking of a two-axis inverted pendulum system: experimental studies.

In this paper, experimental studies of a decentralized neural network control scheme of the reference compensation technique applied to control a 2-degrees-of-freedom (2-DOF) inverted pendulum on an x - y plane are presented. Each axis is controlled by two separate neural network controllers to have a decoupled control structure. Neural network controllers are applied not only to balance the angle of pendulum, but also to control the position tracking of the cart. The decoupled control structure can compensate for uncertainties and cancel coupling effects. Especially, a circular trajectory tracking task is tested for position tracking control of the cart while maintaining the angle of the pendulum. Experimental result shows that position control of the inverted pendulum and cart is successful.

Algicidal activity of thiazolidinedione derivatives against harmful algal blooming species.

Thiazolidinedione (TD) derivatives exhibit algicidal activity against harmful algal blooming species such as Chattonella marina, Heterosigma akashiwo, and Cochlodinium polykrikoides, as reported previously. In this study, the efficacies and selectivities of TD derivatives were tested by analyzing the structure-activity relationships of various TD derivatives. To investigate structure-activity relationships for growth inhibition of harmful algae, we added a methylene group between the cyclohexyl ring and oxygen of 5-(3-chloro-4-hydroxybenzylidene)-TD, which decreased the inhibitory potency of compound 17. Interestingly, another addition of a methylene group significantly increased the inhibitory potency against C. polykrikoides. The addition of 1 µM compound 17 resulted in the cell rupture of harmful algae after less than 10 h incubation at 20 °C. Compound 17 was applied to both harmful and non-harmful algae and showed a drastic reduction in the efficiency of photosystem II, resulting in reduced photosynthetic oxygen evolution. Compound 17 at a 5 µM concentration destroyed all of the harmful algae, while algicidal activity against non-harmful algae did not exceed 30% of the control within the concentration range tested. In contrast, a herbicide, 3-(3,4-dichlorophenyl)-1,1-dimethylurea, tested at a 5 µM concentration, exhibited 40-70% algicidal activity relative to that of the control against both harmful and non-harmful algae. Compound 17 is a promising lead compound for the development of algicides to control harmful algal blooming species.

Enhanced efficacy of TD53, a novel algicidal agent, against the harmful algae via the liposomal delivery system.

The present study aimed to design the liposomal delivery system for TD53, a novel algicial drug in order to improve the delivery properties of TD53 and evaluate its algicidal effects as well as selectivity against harmful and non-harmful algae. Liposomes of TD53 were prepared with 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) by a lyophilization, resulting in relatively small size vesicles (234±38nm) and narrow size distribution (PI=0.130±0.027). The drug leakage from the liposome was negligible in the F/2 media (<2% during 96h incubation). Subsequently algicidal activity of liposomal TD53 against harmful and nonharmful algae was evaluated at various concentrations. The IC(50) values of TD53 in liposome against harmful algae such as Chattonella marina, Heterosigma akashiwo and Cocholodinium polykrikoides were 2.675, 2.029, and 0.480µM, respectively, and were reduced by approximately 50% compared to those obtained from non-liposomal TD53. In contrast, the algicidal effect of liposomal TD53 was insignificant against non-harmful algae including Navicula pelliculosa, Nannochloropsis oculata and Phaeodactylum EPV. Those results suggested that liposomal delivery systems might be effective to enhance the efficacy of TD53 while maintaining the selectivity to harmful algal species.

Electrochemical immunosensor for prostate-specific antigen using self-assembled oligophenylethynylenethiol monolayer containing dendrimer.

Electrochemical immunosensor for prostate-specific antigen (PSA) was fabricated with a self-assembled 4-(2-(4-(acetylthio)phenyl)ethynyl)benzoic acid (APBA) as a bioreceptor. In order to enhance the electrochemical activity of PSA detection, poly(amidoamine) dendrimer was linked on the APBA self-assembled monolayer (SAM). The formation and electrical properties of the SAMs were investigated by surface plasmon resonance and cyclic voltammetry, respectively. The surface morphology of PSA sandwich complex onto the APBA SAM was studied by atomic force microscopy.

Fabrication of self-assembled oligophenylethynylenethiol monolayer for electrochemical glucose biosensor.

An electrochemical glucose biosensor was developed using a gold (Au) electrode, which was composed of self-assembled oligophenylethynylenethiol monolayer and glucose oxidase (GOx) structure. Oligophenylethynylenethiol was used as a chemical linker for the immobilization of GOx on Au electrode, which facilitates the transfer of electron produced by enzyme reaction to the Au electrode. The electrical property of self-assembled oligophenylethynylenethiol monolayer was investigated by using cyclic voltammetry (CV). The formation of self-assembled oligophenylethynylenethiol monolayer and GOx layer on Au surface was verified by using atomic force microscopy (AFM) and surface plasmon resonance (SPR). The electrochemical glucose biosensor exhibited a linear relationship between target concentration and oxidation current in the range of 2-30mM and its detection limit was 2mM.

Assessment of prescribed herbal medicine on liver function in Korea: a prospective observational study.

OBJECTIVE: The aim of this study was to investigate the influence of herbal medicine (HM) prescribed by doctors of Korean medicine (KMD) on liver function in Korea. DESIGN AND INTERVENTIONS: For this multicenter, prospective, observational study, we enrolled patients who wished to take HM prescribed by KMD for various medical purposes in Korea. One hundred and twenty-two (122) patients took HM for an average of 20.6 +/- 8.4 (mean +/- standard deviation) days, and completed questionnaires. OUTCOME MEASURES: Liver function tests (LFTs) were performed before (first test) and after each HM treatment (second test). For LFT, aspartate aminotransferase, alanine aminotransferase, total bilirubin (t-Bil), direct bilirubin, alkaline phosphatase, and gamma-glutamyl transferase were measured. RESULTS: There were no significant changes in LFT data between the first and second tests, except in the t-Bil level. However, all data of total bilirubin level in second test were within normal range, except only one patient. Multivariate analysis did not identify any herb that significantly increased t-Bil; hence no hepatotoxic herb was found. Twenty-one (21) of the 122 patients were abnormal on first testing, and 10 at the second testing. Of the patients taking herbs, 4 changed from normal to abnormal and 15 from abnormal to normal (p = 0.019). CONCLUSION: The current study showed that ingestion of HM prescribed by KMD did not increase the frequency of abnormal LFTs, at least in the short term.