Mitochondrial function contributes to oxysterol-induced osteogenic differentiation in mouse embryonic stem cells

AbstractOxysterols, oxidized derivatives of cholesterol, are biologically active molecules. Specific oxysterols have potent osteogenic properties that act on osteoprogenitor cells. However, the molecular mechanisms underlying these osteoinductive effects on embryonic stem cells (ESCs) are unknown. This study investigated the effect of an oxysterol combination of 22(S)-hydroxycholesterol and 20(S)-hydroxycholesterol (SS) on osteogenic differentiation of ESCs and the alterations to mitochondrial activity during differentiation. Osteogenic differentiation was assessed by alkaline phosphatase (ALP) activity, matrix mineralization, mRNA expression of osteogenic factors, runt-related transcription factor 2, osterix, and osteocalcin, and protein levels of collagen type IA (COLIA) and osteopontin (OPN). Treatment of cells with SS increased osteoinductive activity compared to the control group. Intracellular reactive oxygen species production, intracellular ATP content, mitochondrial membrane potential, mitochondrial mass, mitochondrial DNA copy number, and mRNA expression of peroxisome proliferator-activated receptor-γ coactivators 1α and β, transcription factors involved in mitochondrial biogenesis, were significantly increased during osteogenesis, indicating upregulation of mitochondrial activity. Oxysterol combinations also increased protein levels of mitochondrial respiratory complexes I–V. We also found that SS treatment increased hedgehog signaling target genes, Smo and Gli1 expression. Inhibition of Hh signaling by cyclopamine suppressed mitochondrial biogenesis and ESC osteogenesis. Subsequently, oxysterol-induced Wnt/β-catenin pathways were inhibited by repression of Hh signaling and mitochondrial biogenesis. Transfection of β-catenin specific siRNA decreased the protein levels of COLIA and OPN, as well as ALP activity. Collectively, these data suggest that lipid-based oxysterols enhance differentiation of ESCs toward the osteogenic lineage by regulating mitochondrial activity, canonical Hh/Gli, and Wnt/β-catenin signaling

The Globular Cluster System of M60 (NGC 4649). I. CFHT MOS Spectroscopy and Database

We present the measurement of radial velocities for globular clusters in M60, giant elliptical galaxy in the Virgo cluster. Target globular cluster candidates were selected using the Washington photometry based on the deep 16\arcmin \times 16\arcmin images taken at the KPNO 4m and using the $VI$ photometry derived from the HST/WFPC2 archive images. The spectra of the target objects were obtained using the Multi-Object Spectrograph (MOS) at the Canada-France-Hawaii Telescope (CFHT). We have measured the radial velocity for 111 objects in the field of M60: 93 globular clusters (72 blue globular clusters with $1.0\le(C-T_1)<1.7$ and 21 red globular clusters with $1.7\le(C-T_1)<2.4$), 11 foreground stars, 6 small galaxies, and the nucleus of M60. The measured velocities of the 93 globular clusters range from $\sim 500$ km s$^{-1}$ to $\sim 1600$ km s$^{-1}$, with a mean value of $1070_{-25}^{+27}$ km s$^{-1}$, which is in good agreement with the velocity of the nucleus of M60 ($v_{\rm gal}=1056$ km s$^{-1}$). Combining our results with data in the literature, we present a master catalog of radial velocities for 121 globular clusters in M60. The velocity dispersion of the globular clusters in the master catalog is found to be $234_{-14}^{+13}$ km s$^{-1}$ for the entire sample, $223_{-16}^{+13}$ km s$^{-1}$ for 83 blue globular clusters, and $258_{-31}^{+21}$ km s$^{-1}$ for 38 red globular clusters.Comment: 29 pages, 8 figures. To appear in Ap

Application and evaluation of the MLVA typing assay for the Brucella abortus strains isolated in Korea

<p>Abstract</p> <p>Background</p> <p>A Brucella eradication program has been executed in Korea. To effectively prevent and control brucellosis, a molecular method for genetic identification and epidemiological trace-back must be established. As part of that, the MLVA typing assay was evaluated and applied to <it>B. abortus </it>isolates for analyzing the characteristics of the regional distribution and relationships of foreign isolates.</p> <p>Results</p> <p>A total of 177 isolates originating from 105 cattle farms for the period 1996 to 2008 were selected as representatives for the nine provinces of South Korea. A dendrogram of strain relatedness was constructed in accordance with the number of tandem repeat units for 17 loci so that it was possible to trace back in the restricted areas. Even in a farm contaminated by one source, however, the <it>Brucella </it>isolates showed an increase or decrease in one TRs copy number at some loci with high DI values. Moreover, those 17 loci was confirmed in stability via <it>in-vitro </it>and <it>in-vivo </it>passage, and found to be sufficiently stable markers that can readily identify the inoculated strain even if minor changes were detected. In the parsimony analysis with foreign <it>Brucella </it>isolates, domestic isolates were clustered distinctively, and located near the Central and Southern American isolates.</p> <p>Conclusion</p> <p>The MLVA assay has enough discrimination power in the <it>Brucella </it>species level and can be utilized as a tool for the epidemiological trace-back of the <it>B. abortus </it>isolates. But it is important to consider that <it>Brucella </it>isolates may be capable of undergoing minor changes at some loci in the course of infection or in accordance with the changes of the host.</p

Relationship between Mediterranean diet and depression in South Korea: the Korea National Health and Nutrition Examination Survey

BackgroundSeveral studies have shown that adherence to the Mediterranean diet is associated with a lower risk of depression; however, little is known about the Asian population. This study investigated the relationship between adherence to the Mediterranean diet and depression in a sample of the South Korean population.MethodsIn total, 5,849 adults from the 2014 and 2016 Korea National Health and Nutrition Examination Surveys were included in the study. The Mediterranean diet adherence was measured using a modified alternate Mediterranean diet score (mMED) developed to adjust for Korean dietary patterns. The mMED scores using the Food Frequency Questionnaire were divided into four categories (0–2, 3–4, 5–6, and 7–9 points). Subjects with depression were defined as having moderate-to-severe depressive symptoms using the Patient Health Questionnaire-9, with a cutoff value of 10. Logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). A subgroup analysis was performed based on sex.ResultsThe results of logistic regression analysis indicated that individuals with higher mMED were 42–73% less likely to report depression compared to individuals with the lowest mMED [ORs (95% CIs) =0.58 (0.37–0.90), 0.50 (0.31–0.80), 0.27 (0.15–0.47)] after adjusting for socio-demographic and health-related variables. In women, individuals with mMED of 7–9 had 71% lower odds of depression [ORs (95% CIs): 0.29 (0.13–0.64)]. In men, individuals with mMED of 5–9 had 55% [ORs (95% CIs): 0.45 (0.23–0.91)] to 79% [ORs (95% CIs): 0.21 (0.08–0.57)] lower odds of depression.ConclusionThis study suggests that adherence to the Mediterranean diet is inversely associated with depression in both men and women among Korean adults. This study provides evidence that a Mediterranean diet is crucial in preventing depressive symptoms in Asian populations

The Globular Cluster System of M60 (NGC 4649). II. Kinematics of the Globular Cluster System

We present a kinematic analysis of the globular cluster (GC) system in the giant elliptical galaxy (gE) M60 in the Virgo cluster. Using the photometric and spectroscopic database of 121 GCs (83 blue GCs and 38 red GCs), we have investigated the kinematics of the GC system. We have found that the M60 GC system shows a significant overall rotation. The rotation amplitude of the blue GCs is slightly smaller than or similar to that of the red GCs, and their angles of rotation axes are similar. The velocity dispersions about the mean velocity and about the best fit rotation curve for the red GCs are marginally larger than those for the blue GCs. Comparison of observed stellar and GC velocity dispersion profiles with those calculated from the stellar mass profile shows that the mass-to-light ratio should be increased as the galactocentric distance increases, indicating the existence of an extended dark matter halo. The entire sample of GCs in M60 is found to have a tangentially biased velocity ellipsoid unlike the GC systems in other gEs. Two subsamples appear to have different velocity ellipsoids. The blue GC system has a modest tangentially biased velocity ellipsoid, while the red GC system has a modest radially biased or an isotropic velocity ellipsoid. From the comparison of the kinematic properties of the M60 GC system to those of other gEs (M87, M49, NGC 1399, NGC 5128, and NGC 4636), it is found that the velocity dispersion of the blue GC system is similar to or larger than that of the red GC system except for M60, and the rotation of the GC system is not negligible. The entire sample of each GC system shows an isotropic velocity ellipsoid except for M60, while the subsamples show diverse velocity ellipsoids. We discuss the implication of these results for the formation models of the GC system in gEs.Comment: 48 pages, 16 figures. To appear in Ap

Gene Expression Profile of the Hypothalamus in DNP-KLH Immunized Mice Following Electroacupuncture Stimulation

Clinical evidence indicates that electroacupuncture (EA) is effective for allergic disorder. Recent animal studies have shown that EA treatment reduces levels of IgE and Th2 cytokines in BALB/c mice immunized with 2,4-dinitrophenylated keyhole limpet protein (DNP-KLH). The hypothalamus, a brain center of the neural-immune system, is known to be activated by EA stimulation. This study was performed to identify and characterize the differentially expressed genes in the hypothalamus of DNP-KLH immunized mice that were stimulated with EA or only restrained. To this aim, we conducted a microarray analysis to evaluate the global gene expression profiles, using the hypothalamic RNA samples taken from three groups of mice: (i) normal control group (no treatments); (ii) IMH group (DNP-KLH immunization + restraint); and (iii) IMEA group (immunization + EA stimulation). The microarray analysis revealed that total 39 genes were altered in their expression levels by EA treatment. Ten genes, including T-cell receptor alpha variable region family 13 subfamily 1 (Tcra-V13.1), heat shock protein 1B (Hspa1b) and 2′–5′ oligoadenylate synthetase 1F (Oas1f), were up-regulated in the IMEA group when compared with the IMH group. In contrast, 29 genes, including decay accelerating factor 2 (Daf2), NAD(P)H dehydrogenase, quinone 1 (Nqo1) and programmed cell death 1 ligand 2 (Pdcd1lg2) were down-regulated in the IMEA group as compared with the IMH group. These results suggest that EA treatment can modulate immune response in DNP-KLH immunized mice by regulating expression levels of genes that are associated with innate immune, cellular defense and/or other kinds of immune system in the hypothalamus

Embryoid body size-mediated differential endodermal and mesodermal differentiation using polyethylene glycol (PEG) microwell array

Embryoid bodies have a number of similarities with cells in gastrulation, which provides useful biological information about embryonic stem cell differentiation. Extensive research has been done to study the control of embryoid body-mediated embryonic stem cell differentiation in various research fields. Recently, microengineering technology has been used to control the size of embryoid bodies and to direct lineage specific differentiation of embryonic stem cells. However, the underlying biology of developmental events in the embryoid bodies of different sizes has not been well elucidated. In this study, embryoid bodies with different sizes were generated within microfabricated PEG microwell arrays, and a series of gene and molecular expressions related to early developmental events was investigated to further elucidate the size-mediated differentiation. The gene and molecular expression profile suggested preferential visceral endoderm formation in 450 μm embryoid bodies and preferential lateral plate mesoderm formation in 150 μm embryoid bodies. These aggregates resulted in higher cardiac differentiation in 450 μm embryoid bodies and higher endothelial differentiation in 150 μm embryoid bodies, respectively. Our findings may provide further insight for understanding embryoid body size-mediated developmental progress.National Science Foundation (U.S.) (CAREER Award DMR0847287)United States. Office of Naval Research (Naval Research Young National Investigator Award)National Institutes of Health (U.S.) (HL092836, EB02597, AR057837