Characterization of the ATP transporter in the reconstituted rough endoplasmic reticulum proteoliposomes

AbstractAdenosine triphosphate (ATP) transporter from rat liver rough endoplasmic reticulum (RER) was solubilized and reconstituted into phosphatidylcholine liposomes. The RER proteoliposomes, resulting from optimizing some reconstitution parameters, had an apparent Km value of 1.5 μM and a Vmax of 286 pmol min−1 (mg protein)−1 and showed higher affinity for ATP and a lower Vmax value than intact RER (Km of 6.5 μM and Vmax of 1 nmol). ATP transport was time- and temperature-dependent, inhibited by 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid, which is known as an inhibitor of anion transporters including ATP transporter, but was not affected by atractyloside, a specific inhibitor of mitochondrial ADP/ATP carrier. The internal and external effects of various nucleotides on the ATP transport were examined. ATP transport was cis-inhibited strongly by ADP and weakly by AMP. ADP-preloaded RER proteoliposomes showed a specific increase of ATP transport activity while AMP-preloaded RER proteoliposomes did not show the enhanced overshoot peak in the ATP uptake plot. These results demonstrate the ADP/ATP antiport mechanism of ATP transport in rat liver RER

Human CD57(+ )germinal center-T cells are the major helpers for GC-B cells and induce class switch recombination

BACKGROUND: The function of CD57(+ )CD4(+ )T cells, constituting a major subset of germinal center T (GC-Th) cells in human lymphoid tissues, has been unclear. There have been contradictory reports regarding the B cell helping function of CD57(+ )GC-Th cells in production of immunoglobulin (Ig). Furthermore, the cytokine and co-stimulation requirement for their helper activity remains largely unknown. To clarify and gain more insight into their function in helping B cells, we systematically investigated the capacity of human tonsil CD57(+ )GC-Th cells in inducing B cell Ig synthesis. RESULTS: We demonstrated that CD57(+ )GC-Th cells are highly efficient in helping B cell production of all four subsets of Ig (IgM, IgG, IgA and IgE) compared to other T-helper cells located in germinal centers or interfollicular areas. CD57(+ )GC-Th cells were particularly more efficient than other T cells in helping GC-B cells but not naïve B cells. CD57(+ )GC-Th cells induced the expression of activation-induced cytosine deaminase (AID) and class switch recombination in developing B cells. IgG1-3 and IgA1 were the major Ig isotypes induced by CD57(+ )GC-Th cells. CD40L, but not IL-4, IL-10 and IFN-γ, was critical in CD57(+ )GC-Th cell-driven B cell production of Ig. However, IL-10, when added exogenously, significantly enhanced the helper activity of CD57(+ )GC-Th cells, while TGF-β1 completely and IFN-γ partially suppressed the CD57(+ )GC-Th cell-driven Ig production. CONCLUSIONS: CD57(+)CD4(+ )T cells in the germinal centers of human lymphoid tissues are the major T helper cell subset for GC-B cells in Ig synthesis. Their helper activity is consistent with their capacity to induce AID and class switch recombination, and can be regulated by CD40L, IL-4, IL-10 and TGF-β

Alternative Splicing of the Basic Helix–Loop–Helix Transcription Factor Gene CmbHLH2 Affects Anthocyanin Biosynthesis in Ray Florets of Chrysanthemum (Chrysanthemum morifolium)

Chrysanthemum is an important ornamental crop worldwide. Some white-flowered chrysanthemum cultivars produce red ray florets under natural cultivation conditions, but little is known about how this occurs. We compared the expression of anthocyanin biosynthetic and transcription factor genes between white ray florets and those that turned red based on cultivation conditions to comprehend the underlying mechanism. Significant differences in the expression of CmbHLH2 were detected between the florets of different colors. CmbHLH2 generated two alternatively spliced transcripts, designated CmbHLH2Full and CmbHLH2Short. Compared with CmbHLH2Full, CmbHLH2Short encoded a truncated protein with only a partial MYB-interaction region and no other domains normally present in the full-length protein. Unlike the full-length form, the splicing variant protein CmbHLH2Short localized to the cytoplasm and the nucleus and could not interact with CmMYB6. Additionally, CmbHLH2Short failed to activate anthocyanin biosynthetic genes and induce pigment accumulation in transiently transfected tobacco leaves, whereas CmbHLH2Full promoted both processes when simultaneously expressed with CmMYB6. Co-expressing CmbHLH2Full and CmMYB6 also enhanced the promoter activities of CmCHS and CmDFR. Notably, the Arabidopsis tt8-1 mutant, which lacks red pigmentation in the leaves and seeds, could be complemented by the heterologous expression of CmbHLH2Full, which restored red pigmentation and resulted in red pigmentation in high anthocyanin and proanthocyanidin contents in the leaves and seeds, respectively, whereas expression of CmbHLH2Short did not. Together, these results indicate that CmbHLH2 and CmMYB6 interaction plays a key role in the anthocyanin pigmentation changes of ray florets in chrysanthemum. Our findings highlight alternative splicing as a potential approach to modulate anthocyanin biosynthesis in specific tissues

Satellite Laser Ranging System at Geochang Sta

Korea Astronomy and Space Science Institute (KASI) has been developing the space optical and laser tracking (SOLT) system for space geodesy, space situational awareness, and Korean space missions. The SOLT system comprises satellite laser ranging (SLR), adaptive optics (AO), and debris laser tracking (DLT) systems, which share numerous subsystems, such as an optical telescope and tracking mount. It is designed to be capable of laser ranging up to geosynchronous Earth orbit satellites with a laser retro-reflector array, space objects imaging brighter than magnitude 10, and laser tracking low Earth orbit space debris of uncooperative targets. For the realization of multiple functions in a novel configuration, the SOLT system employs a switching mirror that is installed inside the telescope pedestal and feeds the beam path to each system. The SLR and AO systems have already been established at the Geochang station, whereas the DLT system is currently under development and the AO system is being prepared for testing. In this study, the design and development of the SOLT system are addressed and the SLR data quality is evaluated compared to the International Laser Ranging Service (ILRS) tracking stations in terms of single-shot ranging precision. The analysis results indicate that the SLR system has a good ranging performance, to a few millimeters precision. Therefore, it is expected that the SLR system will not only play an important role as a member of the ILRS tracking network, but also contribute to future Korean space missions

The mid-infrared view of red sequence galaxies in Abell 2218 with <i>AKARI</i>

We present the AKARI Infrared Camera (IRC) imaging observation of early-type galaxies (ETGs) in A2218 at z ~ 0.175. Mid-infrared (MIR) emission from ETGs traces circumstellar dust emission from asymptotic giant branch (AGB) stars or/and residual star formation. Including the unique imaging capability at 11 and 15 μm, our AKARI data provide an effective way to investigate MIR properties of ETGs in the cluster environment. Among our flux-limited sample of 22 red sequence ETGs with precise dynamical and line strength measurements (less than 18 mag at 3 μm), we find that at least 41% have MIR-excess emission. The N3 – S11 versus N3 (3 and 11 μm) color-magnitude relation shows the expected blue sequence, but the MIR-excess galaxies add a red wing to the relation especially at the fainter end. A spectral energy distribution analysis reveals that the dust emission from AGB stars is the most likely cause of the MIR excess, with a low level of star formation being the next possible explanation. The MIR-excess galaxies show a wide spread of N3 – S11 colors, implying a significant spread (2-11 Gyr) in the estimated mean ages of stellar populations. We study the environmental dependence of MIR-excess ETGs over an area out to a half virial radius (~1 Mpc). We find that the MIR-excess ETGs are preferentially located in the outer region. From this evidence, we suggest that the fainter, MIR-excess ETGs have just joined the red sequence, possibly due to the infall and subsequent morphological/spectral transformation induced by the cluster environment

Determination of the Distal Fusion Level in the Management of Thoracolumbar and Lumbar Adolescent Idiopathic Scoliosis Using Pedicle Screw Instrumentation

Study DesignA retrospective study.PurposeTo determine the exact distal fusion level in the management of thoracolumbar/lumbar adolescent idiopathic scoliosis (TL/L AIS) using pedicle screw instrumentation (PSI).Overview of LiteratureThe selection of distal fusion level remains controversial in TL/L AIS.MethodsRadiographic parameters of 66 TL/L AIS patients were analyzed. The patients were grouped according to the distal fusion level; L3 group (fusion to L3, n=58) and L4 group (fusion to L4, n=8). The L3 group was subdivided into L3A (L3 crosses the mid-sacral line with rotation of less than grade II, n=33) and L3B (L3 does not cross the mid-sacral line or rotation is grade II or more, n=25) based on both bending radiographs. All of the patients in the L4 group had the same location and rotation of L3 in bending films as that of patients in the L3B group. An unsatisfactory result was defined as a lowest instrumented vertebral tilt (LIVT) of more than 10° or coronal balance of more than 15 mm.ResultsAmong the 3 groups, there was a significantly lesser correction in the TL/L curve and LIVT in the L3B group. Unsatisfactory results were obtained in 3 patients (9.1%) of the L3A group, in 15 patients (68.2%) of the L3B group, and in 1 patient (12.5%) of the L4 group with a significant difference.ConclusionsIn TL/L AIS treatment with PSI, the curve can be fused to L3 with favorable radiographic outcomes when L3 crosses the mid-sacral line with rotation of less than grade II in bending films. Otherwise, fusion has to be extended to L4