Science.gov

Sample records for 7a 7b 7c

  1. MicroRNAs as biomarkers for major depression: a role for let-7b and let-7c

    PubMed Central

    Gururajan, A; Naughton, M E; Scott, K A; O'Connor, R M; Moloney, G; Clarke, G; Dowling, J; Walsh, A; Ismail, F; Shorten, G; Scott, L; McLoughlin, D M; Cryan, J F; Dinan, T G

    2016-01-01

    There is a growing emphasis in the field of psychiatry on the need to identify candidate biomarkers to aid in diagnosis and clinical management of depression, particularly with respect to predicting response to specific therapeutic strategies. MicroRNAs are small nucleotide sequences with the ability to regulate gene expression at the transcriptomic level and emerging evidence from a range of studies has highlighted their biomarker potential. Here we compared healthy controls (n=20) with patients diagnosed with major depression (n=40) and who were treatment-resistant to identify peripheral microRNA biomarkers, which could be used for diagnosis and to predict response to electroconvulsive therapy (ECT) and ketamine (KET) infusions, treatments that have previously shown to be effective in treatment-resistant depression (TRD). At baseline and after treatment, blood samples were taken and symptom severity scores rated using the Hamilton Depression Rating Scale (HDRS). Samples were analyzed for microRNA expression using microarray and validated using quantitative PCR. As expected, both treatments reduced HDRS scores. Compared with controls, the baseline expression of the microRNA let-7b was less by ~40% in TRD patients compared with controls. The baseline expression of let-7c was also lower by ~50% in TRD patients who received ECT. Bioinformatic analysis revealed that let-7b and let-7c regulates the expression of 27 genes in the PI3k-Akt-mTOR signaling pathway, which has previously been reported to be dysfunctional in depression. The expression of miR-16, miR-182, miR-451 and miR-223 were similar to that in controls. Baseline microRNA expression could not predict treatment response and microRNAs were unaffected by treatment. Taken together, we have identified let-7b and let-7c as candidate biomarkers of major depression. PMID:27483380

  2. MicroRNAs as biomarkers for major depression: a role for let-7b and let-7c.

    PubMed

    Gururajan, A; Naughton, M E; Scott, K A; O'Connor, R M; Moloney, G; Clarke, G; Dowling, J; Walsh, A; Ismail, F; Shorten, G; Scott, L; McLoughlin, D M; Cryan, J F; Dinan, T G

    2016-01-01

    There is a growing emphasis in the field of psychiatry on the need to identify candidate biomarkers to aid in diagnosis and clinical management of depression, particularly with respect to predicting response to specific therapeutic strategies. MicroRNAs are small nucleotide sequences with the ability to regulate gene expression at the transcriptomic level and emerging evidence from a range of studies has highlighted their biomarker potential. Here we compared healthy controls (n=20) with patients diagnosed with major depression (n=40) and who were treatment-resistant to identify peripheral microRNA biomarkers, which could be used for diagnosis and to predict response to electroconvulsive therapy (ECT) and ketamine (KET) infusions, treatments that have previously shown to be effective in treatment-resistant depression (TRD). At baseline and after treatment, blood samples were taken and symptom severity scores rated using the Hamilton Depression Rating Scale (HDRS). Samples were analyzed for microRNA expression using microarray and validated using quantitative PCR. As expected, both treatments reduced HDRS scores. Compared with controls, the baseline expression of the microRNA let-7b was less by ~40% in TRD patients compared with controls. The baseline expression of let-7c was also lower by ~50% in TRD patients who received ECT. Bioinformatic analysis revealed that let-7b and let-7c regulates the expression of 27 genes in the PI3k-Akt-mTOR signaling pathway, which has previously been reported to be dysfunctional in depression. The expression of miR-16, miR-182, miR-451 and miR-223 were similar to that in controls. Baseline microRNA expression could not predict treatment response and microRNAs were unaffected by treatment. Taken together, we have identified let-7b and let-7c as candidate biomarkers of major depression. PMID:27483380

  3. Metal-Dependent Regulation of ATP7A and ATP7B in Fibroblast Cultures

    PubMed Central

    Lenartowicz, Malgorzata; Moos, Torben; Ogórek, Mateusz; Jensen, Thomas G.; Møller, Lisbeth B.

    2016-01-01

    Deficiency of one of the copper transporters ATP7A and ATP7B leads to the rare X-linked disorder Menkes Disease (MD) or the rare autosomal disorder Wilson disease (WD), respectively. In order to investigate whether the ATP7A and the ATP7B genes may be transcriptionally regulated, we measured the expression level of the two genes at various concentrations of iron, copper, and insulin. Treating fibroblasts from controls or from individuals with MD or WD for 3 and 10 days with iron chelators revealed that iron deficiency led to increased transcript levels of both ATP7A and ATP7B. Copper deficiency obtained by treatment with the copper chelator led to a downregulation of ATP7A in the control fibroblasts, but surprisingly not in the WD fibroblasts. In contrast, the addition of copper led to an increased expression of ATP7A, but a decreased expression of ATP7B. Thus, whereas similar regulation patterns for the two genes were observed in response to iron deficiency, different responses were observed after changes in the access to copper. Mosaic fibroblast cultures from female carriers of MD treated with copper or copper chelator for 6–8 weeks led to clonal selection. Cells that express the normal ATP7A allele had a selective growth advantage at high copper concentrations, whereas more surprisingly, cells that express the mutant ATP7A allele had a selective growth advantage at low copper concentrations. Thus, although the transcription of ATP7A is regulated by copper, clonal growth selection in mosaic cell cultures is affected by the level of copper. Female carriers of MD are rarely affected probably due to a skewed inactivation of the X-chromosome bearing the ATP7A mutation. PMID:27587995

  4. Metal-Dependent Regulation of ATP7A and ATP7B in Fibroblast Cultures.

    PubMed

    Lenartowicz, Malgorzata; Moos, Torben; Ogórek, Mateusz; Jensen, Thomas G; Møller, Lisbeth B

    2016-01-01

    Deficiency of one of the copper transporters ATP7A and ATP7B leads to the rare X-linked disorder Menkes Disease (MD) or the rare autosomal disorder Wilson disease (WD), respectively. In order to investigate whether the ATP7A and the ATP7B genes may be transcriptionally regulated, we measured the expression level of the two genes at various concentrations of iron, copper, and insulin. Treating fibroblasts from controls or from individuals with MD or WD for 3 and 10 days with iron chelators revealed that iron deficiency led to increased transcript levels of both ATP7A and ATP7B. Copper deficiency obtained by treatment with the copper chelator led to a downregulation of ATP7A in the control fibroblasts, but surprisingly not in the WD fibroblasts. In contrast, the addition of copper led to an increased expression of ATP7A, but a decreased expression of ATP7B. Thus, whereas similar regulation patterns for the two genes were observed in response to iron deficiency, different responses were observed after changes in the access to copper. Mosaic fibroblast cultures from female carriers of MD treated with copper or copper chelator for 6-8 weeks led to clonal selection. Cells that express the normal ATP7A allele had a selective growth advantage at high copper concentrations, whereas more surprisingly, cells that express the mutant ATP7A allele had a selective growth advantage at low copper concentrations. Thus, although the transcription of ATP7A is regulated by copper, clonal growth selection in mosaic cell cultures is affected by the level of copper. Female carriers of MD are rarely affected probably due to a skewed inactivation of the X-chromosome bearing the ATP7A mutation. PMID:27587995

  5. TRIPOLI-4 criticality calculations for MOX fuelled SNEAK 7A and 7B fast critical assemblies

    SciTech Connect

    Lee, Y. K.

    2012-07-01

    A prototype Generation IV fast neutron reactor is under design and development in France. The MOX fuel will be introduced into this self-generating core in order to demonstrate low net plutonium production. To support the TRIPOLI-4 Monte Carlo transport code in criticality calculations of fast reactors, the effective delayed neutron fraction {beta}eff estimation and the Probability Tables (PT) option to treat the unresolved resonance region of cross-sections are two essentials. In this study, TRIPOLI-4 calculations have been made using current nuclear data libraries JEFF-3.1.1 and ENDF/B-VII.0 to benchmark the reactor physics parameters of the MOX fuelled SNEAK 7A and 7B fast critical assemblies. TRIPOLI-4 calculated K{sub eff} and {beta}eff of the homogeneous R-Z models and the 3D multi-cell models have been validated against the measured ones. The impact of the PT option on K{sub eff} is 340 {+-} 10 pcm for SNEAK 7A core and 410 {+-} 12 pcm for 7B. Four-group spectra and energy spectral indices, f8/f5, f9/f5, and c8/f5 in the two SNEAK cores have also been calculated with the TRIPOLI-4 mesh tally. Calculated spectrum-hardening index f8/f5 is 0.0418 for SNEAK 7A and 0.0315 for 7B. From this study the SNEAK 3D models have been verified for the next revision of IRPhE (International Handbook of Evaluated Reactor Physics Benchmark Experiments). (authors)

  6. Clusterin and COMMD1 Independently Regulate Degradation of the Mammalian Copper ATPases ATP7A and ATP7B*

    PubMed Central

    Materia, Stephanie; Cater, Michael A.; Klomp, Leo W. J.; Mercer, Julian F. B.; La Fontaine, Sharon

    2012-01-01

    ATP7A and ATP7B are copper-transporting P1B-type ATPases (Cu-ATPases) that are critical for regulating intracellular copper homeostasis. Mutations in the genes encoding ATP7A and ATP7B lead to copper deficiency and copper toxicity disorders, Menkes and Wilson diseases, respectively. Clusterin and COMMD1 were previously identified as interacting partners of these Cu-ATPases. In this study, we confirmed that clusterin and COMMD1 interact to down-regulate both ATP7A and ATP7B. Overexpression and knockdown of clusterin/COMMD1 decreased and increased, respectively, endogenous levels of ATP7A and ATP7B, consistent with a role in facilitating Cu-ATPase degradation. We demonstrate that whereas the clusterin/ATP7B interaction was enhanced by oxidative stress or mutation of ATP7B, the COMMD1/ATP7B interaction did not change under oxidative stress conditions, and only increased with ATP7B mutations that led to its misfolding. Clusterin and COMMD1 facilitated the degradation of ATP7B containing the same Wilson disease-causing C-terminal mutations via different degradation pathways, clusterin via the lysosomal pathway and COMMD1 via the proteasomal pathway. Furthermore, endogenous ATP7B existed in a complex with clusterin and COMMD1, but these interactions were neither competitive nor cooperative and occurred independently of each other. Together these data indicate that clusterin and COMMD1 represent alternative and independent systems regulating Cu-ATPase quality control, and consequently contributing to the maintenance of copper homeostasis. PMID:22130675

  7. Syntrophic biodegradation of butachlor by Mycobacterium sp. J7A and Sphingobium sp. J7B isolated from rice paddy soil.

    PubMed

    Kim, Nam Hyun; Kim, Dong-Uk; Kim, Ijung; Ka, Jong-Ok

    2013-07-01

    Two bacterial strains involved in syntrophic degradation of chloroacetamide herbicide butachlor were isolated from a rice paddy soil. Analysis of 16S rRNA gene sequences indicated that the two isolates were related to members of the genera Mycobacterium and Sphingobium, respectively. Thus, a pair consisted of Mycobacterium sp. J7A and Sphingobium sp. J7B could rapidly degrade butachlor (100 mg L(-1)) at 28 °C within 24 h, while each isolate alone was not able to completely degrade butachlor. The isolate Mycobacterium sp. J7A was observed to grow slightly on butachlor, possibly utilizing the alkyl side chain of butachlor as its carbon and energy source, but the isolate Sphingobium sp. J7B alone could not grow on butachlor at all. Gas chromatography-mass spectrometry on catabolic intermediates revealed that the strain J7A produced and accumulated 2-chloro-N-(2,6-diethylphenyl) acetamide (CDEPA) during growth on butachlor. This intermediate was not further degraded by strain J7A, but strain J7B was observed to be able to completely degrade and grow on it through 2,6-diethylaniline (DEA). The results showed that butachlor was completely degraded by the two isolates by syntrophic metabolism, in which strain Mycobacterium sp. J7A degraded butachlor to CDEPA, which was subsequently degraded by strain Sphingobium sp. J7B through DEA.

  8. Altered localisation of the copper efflux transporters ATP7A and ATP7B associated with cisplatin resistance in human ovarian carcinoma cells

    PubMed Central

    Kalayda, Ganna V; Wagner, Christina H; Buß, Irina; Reedijk, Jan; Jaehde, Ulrich

    2008-01-01

    Background Copper homeostasis proteins ATP7A and ATP7B are assumed to be involved in the intracellular transport of cisplatin. The aim of the present study was to assess the relevance of sub cellular localisation of these transporters for acquired cisplatin resistance in vitro. For this purpose, localisation of ATP7A and ATP7B in A2780 human ovarian carcinoma cells and their cisplatin-resistant variant, A2780cis, was investigated. Methods Sub cellular localisation of ATP7A and ATP7B in sensitive and resistant cells was investigated using confocal fluorescence microscopy after immunohistochemical staining. Co-localisation experiments with a cisplatin analogue modified with a carboxyfluorescein-diacetate residue were performed. Cytotoxicity of the fluorescent cisplatin analogue in A2780 and A2780cis cells was determined using an MTT-based assay. The significance of differences was analysed using Student's t test or Mann-Whitney test as appropriate, p values of < 0.05 were considered significant. Results In the sensitive cells, both transporters are mainly localised in the trans-Golgi network, whereas they are sequestrated in more peripherally located vesicles in the resistant cells. Altered localisation of ATP7A and ATP7B in A2780cis cells is likely to be a consequence of major abnormalities in intracellular protein trafficking related to a reduced lysosomal compartment in this cell line. Changes in sub cellular localisation of ATP7A and ATP7B may facilitate sequestration of cisplatin in the vesicular structures of A2780cis cells, which may prevent drug binding to genomic DNA and thereby contribute to cisplatin resistance. Conclusion Our results indicate that alterations in sub cellular localisation of transport proteins may contribute to cisplatin resistance in vitro. Investigation of intracellular protein localisation in primary tumour cell cultures and tumour tissues may help to develop markers of clinically relevant cisplatin resistance. Detection of resistant

  9. Iowa Commercial Pesticide Applicator Manual, Category 7A: General and Household Pest Control. CS-19. Category 7B: Termite Control, CS-20. Category 7C: Food Industry Pest Control, CS-21. Category 7D: Community Insect Control, CS-22.

    ERIC Educational Resources Information Center

    Stockdale, Harold J., Ed.; And Others

    This manual provides information needed to meet specific standards for certification as a pesticide applicator. The first section discusses general and household pest control and is concerned with parasitic pests and man, stored product pests, and irritating vertebrates. Section two is devoted to identifying and controlling structural pests such…

  10. Expression of the copper transporters hCtr1, ATP7A and ATP7B is associated with the response to chemotherapy and survival time in patients with resected non-small cell lung cancer

    PubMed Central

    YANG, TIAN; CHEN, MINGWEI; CHEN, TIANJUN; THAKUR, ASMITANANDA

    2015-01-01

    Copper transporter family proteins may regulate the chemoresistance of non-small cell lung cancer (NSCLC) to platinum-based anticancer drugs. The present study aimed to investigate the expression of these proteins in lung cancer tissue specimens for association with clinicopathological data and patient responses to chemotherapy and survival. A total of 54 patients with surgically resected NSCLC that received first-line platinum-based doublet chemotherapy were recruited in the present study, and the paraffin-embedded pre-treatment tumor tissue specimens were subjected to immunohistochemical analysis for the expression of human copper transporter 1 (hCtr1) and copper-transporting p-type adenosine triphosphatase 1 (ATP7A) and 2 (ATP7B). This cohort of patients with NSCLC received platinum-based chemotherapy subsequent to the surgical removal of tumor lesions. ATP7B expression was found to be significantly associated with tumor cell differentiation, while hCtr1 expression was significantly associated with improved chemotherapeutic responses. The median survival time was 20 months in patients possessing tumors with high ATP7A expression, but >66 months in patients possessing tumors with low ATP7A expression at the end of the follow-up (P<0.001). The median survival time at the end of the follow-up was 15 months in patients with low tumor hCtr1 expression, but >66 months in patients with high tumor hCtr1 expression (P<0.001). High hCtr1 and low ATP7A expression were each favorable prognostic factors subsequent to chemotherapy for patients with resected NSCLC. Multivariate analysis revealed that high hCtr1 expression combined with low ATP7A expression, good tumor differentiation and female gender were all favorable independent predictive and prognostic factors for patients with resected NSCLC following chemotherapy. High hCtr1 expression combined with low ATP7A expression was associated with an improved prognosis in patients with resected NSCLC that received platinum

  11. The Tumor Suppressor BCL7B Functions in the Wnt Signaling Pathway.

    PubMed

    Uehara, Tomoko; Kage-Nakadai, Eriko; Yoshina, Sawako; Imae, Rieko; Mitani, Shohei

    2015-01-01

    Human BCL7 gene family consists of BCL7A, BCL7B, and BCL7C. A number of clinical studies have reported that BCL7 family is involved in cancer incidence, progression, and development. Among them, BCL7B, located on chromosome 7q11.23, is one of the deleted genes in patients with Williams-Beuren syndrome. Although several studies have suggested that malignant diseases occurring in patients with Williams-Beuren syndrome are associated with aberrations in BCL7B, little is known regarding the function of this gene at the cellular level. In this study, we focused on bcl-7, which is the only homolog of BCL7 gene family in Caenorhabditis elegans, and analyzed bcl-7 deletion mutants. As a result, we found that bcl-7 is required for the asymmetric differentiation of epithelial seam cells, which have self-renewal properties as stem cells and divide asymmetrically through the WNT pathway. Distal tip cell development, which is regulated by the WNT pathway in Caenorhabditis elegans, was also affected in bcl-7-knockout mutants. Interestingly, bcl-7 mutants exhibited nuclear enlargement, reminiscent of the anaplastic features of malignant cells. Furthermore, in KATOIII human gastric cancer cells, BCL7B knockdown induced nuclear enlargement, promoted the multinuclei phenotype and suppressed cell death. In addition, this study showed that BCL7B negatively regulates the Wnt-signaling pathway and positively regulates the apoptotic pathway. Taken together, our data indicate that BCL7B/BCL-7 has some roles in maintaining the structure of nuclei and is involved in the modulation of multiple pathways, including Wnt and apoptosis. This study may implicate a risk of malignancies with BCL7B-deficiency, such as Williams-Beuren syndrome. PMID:25569233

  12. The Tumor Suppressor BCL7B Functions in the Wnt Signaling Pathway

    PubMed Central

    Uehara, Tomoko; Kage-Nakadai, Eriko; Yoshina, Sawako; Imae, Rieko; Mitani, Shohei

    2015-01-01

    Human BCL7 gene family consists of BCL7A, BCL7B, and BCL7C. A number of clinical studies have reported that BCL7 family is involved in cancer incidence, progression, and development. Among them, BCL7B, located on chromosome 7q11.23, is one of the deleted genes in patients with Williams-Beuren syndrome. Although several studies have suggested that malignant diseases occurring in patients with Williams-Beuren syndrome are associated with aberrations in BCL7B, little is known regarding the function of this gene at the cellular level. In this study, we focused on bcl-7, which is the only homolog of BCL7 gene family in Caenorhabditis elegans, and analyzed bcl-7 deletion mutants. As a result, we found that bcl-7 is required for the asymmetric differentiation of epithelial seam cells, which have self-renewal properties as stem cells and divide asymmetrically through the WNT pathway. Distal tip cell development, which is regulated by the WNT pathway in Caenorhabditis elegans, was also affected in bcl-7-knockout mutants. Interestingly, bcl-7 mutants exhibited nuclear enlargement, reminiscent of the anaplastic features of malignant cells. Furthermore, in KATOIII human gastric cancer cells, BCL7B knockdown induced nuclear enlargement, promoted the multinuclei phenotype and suppressed cell death. In addition, this study showed that BCL7B negatively regulates the Wnt-signaling pathway and positively regulates the apoptotic pathway. Taken together, our data indicate that BCL7B/BCL-7 has some roles in maintaining the structure of nuclei and is involved in the modulation of multiple pathways, including Wnt and apoptosis. This study may implicate a risk of malignancies with BCL7B-deficiency, such as Williams-Beuren syndrome. PMID:25569233

  13. ATP7B detoxifies silver in ciliated airway epithelial cells

    SciTech Connect

    Ibricevic, Aida; Brody, Steven L.; Youngs, Wiley J.; Cannon, Carolyn L.

    2010-03-15

    Silver is a centuries-old antibiotic agent currently used to treat infected burns. The sensitivity of a wide range of drug-resistant microorganisms to silver killing suggests that it may be useful for treating refractory lung infections. Toward this goal, we previously developed a methylated caffeine silver acetate compound, SCC1, that exhibits broad-spectrum antimicrobial activity against clinical strains of bacteria in vitro and when nebulized to lungs in mouse infection models. Preclinical testing of high concentrations of SCC1 in primary culture mouse tracheal epithelial cells (mTEC) showed selective ciliated cell death. Ciliated cell death was induced by both silver- and copper-containing compounds but not by the methylated caffeine portion of SCC1. We hypothesized that copper transporting P-type ATPases, ATP7A and ATP7B, play a role in silver detoxification in the airway. In mTEC, ATP7A was expressed in non-ciliated cells, whereas ATP7B was expressed only in ciliated cells. The exposure of mTEC to SCC1 induced the trafficking of ATP7B, but not ATP7A, suggesting the presence of a cell-specific silver uptake and detoxification mechanisms. Indeed, the expression of the copper uptake protein CTR1 was also restricted to ciliated cells. A role of ATP7B in silver detoxification was further substantiated when treatment of SCC1 significantly increased cell death in ATP7B shRNA-treated HepG2 cells. In addition, mTEC from ATP7B{sup -/-} mice showed enhanced loss of ciliated cells compared to wild type. These studies are the first to demonstrate a cell type-specific expression of the Ag{sup +}/Cu{sup +} transporters ATP7A, ATP7B, and CTR1 in airway epithelial cells and a role for ATP7B in detoxification of these metals in the lung.

  14. ATP7B expression confers multidrug resistance through drug sequestration.

    PubMed

    Moinuddin, F M; Shinsato, Yoshinari; Komatsu, Masaharu; Mitsuo, Ryoichi; Minami, Kentaro; Yamamoto, Masatatsu; Kawahara, Kohich; Hirano, Hirofumi; Arita, Kazunori; Furukawa, Tatsuhiko

    2016-04-19

    We previously reported that ATP7B is involved in cisplatin resistance and ATP7A confers multidrug resistance (MDR) in cancer cells.In this study, we show that ATP7B expressing cells also are resistant to doxorubicin, SN-38, etoposide, and paclitaxel as well as cisplatin.In ATP7B expressing cells, doxorubicin relocated from the nuclei to the late-endosome at 4 hours after doxorubicin exposure. EGFP-ATP7B mainly colocalized with doxorubicin.ATP7B has six metal binding sites (MBSs) in the N-terminal cytoplasmic region. To investigate the role of the MBSs of ATP7B in doxorubicin resistance, we used three mutant ATP7B (Cu0, Cu6 and M6C/S) expressing cells. Cu0 has no MBSs, Cu6 has only the sixth MBS and M6C/S carries CXXC to SXXS mutation in the sixth MBS. Cu6 expressing cells were less resistance to the anticancer agents than wild type ATP7B expressing cells, and had doxorubicin sequestration in the late-endosome. Cu0- and M6C/S-expressing cells were sensitive to doxorubicin. In these cells, doxorubicin did not relocalize to the late-endosome. EGFP-M6C/S mainly localized to the trans-Golgi network (TGN) even in the presence of copper. Thus the cysteine residues in the sixth MBS of ATP7B are essential for MDR phenotype.Finally, we found that ammonium chloride and tamoxifen suppressed late endosomal sequestration of doxorubicin, thereby attenuating drug resistance. These results suggest that the sequestration depends on the acidity of the vesicles partly.We here demonstrate that ATP7B confers MDR by facilitating nuclear drug efflux and late endosomal drug sequestration. PMID:26988911

  15. ATP7B expression confers multidrug resistance through drug sequestration

    PubMed Central

    Moinuddin, F M; Shinsato, Yoshinari; Komatsu, Masaharu; Mitsuo, Ryoichi; Minami, Kentaro; Yamamoto, Masatatsu; Kawahara, Kohich; Hirano, Hirofumi; Arita, Kazunori; Furukawa, Tatsuhiko

    2016-01-01

    We previously reported that ATP7B is involved in cisplatin resistance and ATP7A confers multidrug resistance (MDR) in cancer cells. In this study, we show that ATP7B expressing cells also are resistant to doxorubicin, SN-38, etoposide, and paclitaxel as well as cisplatin. In ATP7B expressing cells, doxorubicin relocated from the nuclei to the late-endosome at 4 hours after doxorubicin exposure. EGFP-ATP7B mainly colocalized with doxorubicin. ATP7B has six metal binding sites (MBSs) in the N-terminal cytoplasmic region. To investigate the role of the MBSs of ATP7B in doxorubicin resistance, we used three mutant ATP7B (Cu0, Cu6 and M6C/S) expressing cells. Cu0 has no MBSs, Cu6 has only the sixth MBS and M6C/S carries CXXC to SXXS mutation in the sixth MBS. Cu6 expressing cells were less resistance to the anticancer agents than wild type ATP7B expressing cells, and had doxorubicin sequestration in the late-endosome. Cu0- and M6C/S-expressing cells were sensitive to doxorubicin. In these cells, doxorubicin did not relocalize to the late-endosome. EGFP-M6C/S mainly localized to the trans-Golgi network (TGN) even in the presence of copper. Thus the cysteine residues in the sixth MBS of ATP7B are essential for MDR phenotype. Finally, we found that ammonium chloride and tamoxifen suppressed late endosomal sequestration of doxorubicin, thereby attenuating drug resistance. These results suggest that the sequestration depends on the acidity of the vesicles partly. We here demonstrate that ATP7B confers MDR by facilitating nuclear drug efflux and late endosomal drug sequestration. PMID:26988911

  16. Estimated Environmental Exposures for MISSE-7B

    NASA Technical Reports Server (NTRS)

    Finckenor, Miria M.; Moore, Chip; Norwood, Joseph K.; Henrie, Ben; DeGroh, Kim

    2012-01-01

    This paper details the 18-month environmental exposure for Materials International Space Station Experiment 7B (MISSE-7B) ram and wake sides. This includes atomic oxygen, ultraviolet radiation, particulate radiation, thermal cycling, meteoroid/space debris impacts, and observed contamination. Atomic oxygen fluence was determined by measured mass and thickness loss of polymers of known reactivity. Diodes sensitive to ultraviolet light actively measured solar radiation incident on the experiment. Comparisons to earlier MISSE flights are discussed.

  17. SLUDGE BATCH 7B GLASS VARIABILITY STUDY

    SciTech Connect

    Johnson, F.; Edwards, T.

    2011-10-25

    The Defense Waste Processing Facility (DWPF) is preparing to initiate processing Sludge Batch 7b (SB7b). In support of the upcoming processing, the Savannah River National Laboratory (SRNL) provided a recommendation to utilize Frits 418 with a 6% Na{sub 2}O addition (26 wt% Na{sub 2}O in sludge) and 702 with a 4% Na{sub 2}O addition (24 wt% Na{sub 2}O in sludge) to process SB7b. This recommendation was based on assessments of the compositional projections for SB7b available at the time from the Savannah River Remediation (SRR). To support qualification of SB7b, SRNL executed a variability study to assess the applicability of the current durability models for SB7b. The durability models were assessed over the expected composition range of SB7b, including potential caustic additions, combined with Frits 702 and 418 over a 32-40% waste loading (WL) range. Thirty four glasses were selected based on Frits 418 and 702 coupled with the sludge projections with an additional 4-6% Na{sub 2}O to reflect the potential caustic addition. Six of these glasses, based on average nominal sludge compositions including the appropriate caustic addition, were developed for both Frit 418 and Frit 702 at 32, 36 and 40% WL to provide coverage in the center of the anticipated SB7b glass region. All glasses were fabricated and characterized using chemical composition analysis, X-ray diffraction (XRD) and the Product Consistency Test (PCT). To comply with the DWPF Glass Product Control Program, a total of thirty four glasses were fabricated to assess the applicability of the current DWPF PCCS durability models. Based on the measured PCT response, all of the glasses were acceptable with respect to the Environmental Assessment (EA) benchmark glass regardless of thermal history. The NL[B] values of the SB7b variability study glasses were less than 1.99 g/L as compared to 16.695 g/L for EA. A small number of the D-optimally selected 'outer layer' extreme vertices (EV) glasses were not predictable

  18. Analysis Of The Sludge Batch 7b (Macrobatch 9) DWPF Pour Stream Glass Sample

    SciTech Connect

    Johnson, F. C.; Crawford, C. L.; Pareizs, J. M.

    2013-11-18

    The Defense Waste Processing Facility (DWPF) began processing Sludge Batch 7b (SB7b), also referred to as Macrobatch 9 (MB9), in January 2012. SB7b is a blend of the heel of Tank 40 from Sludge Batch 7a (SB7a) and the SB7b material that was transferred to Tank 40 from Tank 51. SB7b was processed using Frit 418. During processing of each sludge batch, the DWPF is required to take at least one glass sample to meet the objectives of the Glass Product Control Program (GPCP), which is governed by the DWPF Waste Form Compliance Plan, and to complete the necessary Production Records so that the final glass product may be disposed of at a Federal Repository. Two pour stream glass samples were collected while processing SB7b. The samples were transferred to the Savannah River National Laboratory (SRNL) where one was analyzed and the other was archived. The following conclusions were drawn from the analytical results provided in this report: The sum of oxides for the official SB7b pour stream glass is within the Product Composition Control System (PCCS) limits (95-105 wt%); The average calculated Waste Dilution Factor (WDF) for SB7b is 2.3. In general, the measured radionuclide content of the official SB7b pour stream glass is in good agreement with the calculated values from the Tank 40 dried sludge results from the SB7b Waste Acceptance Program Specification (WAPS) sample; As in previous pour stream samples, ruthenium and rhodium inclusions were detected by Scanning Electron Microscopy-Electron Dispersive Spectroscopy (SEM-EDS) in the SB7b pour stream sample; The Product Consistency Test (PCT) results indicate that the official SB7b pour stream glass meets the waste acceptance criteria for durability with a normalized boron release of 0.8 g/L, which is an order of magnitude less than the Environmental Assessment (EA) glass; The measured density of the SB7b pour stream glass was 2.70 g/cm{sup 3}; The Fe{sup 2+}/ΣFe ratio of the SB7b pour stream samples was 0.07.

  19. SLUDGE BATCH 7B QUALIFICATION ACTIVITIES WITH SRS TANK FARM SLUDGE

    SciTech Connect

    Pareizs, J.; Click, D.; Lambert, D.; Reboul, S.

    2011-11-16

    Waste Solidification Engineering (WSE) has requested that characterization and a radioactive demonstration of the next batch of sludge slurry - Sludge Batch 7b (SB7b) - be completed in the Shielded Cells Facility of the Savannah River National Laboratory (SRNL) via a Technical Task Request (TTR). This characterization and demonstration, or sludge batch qualification process, is required prior to transfer of the sludge from Tank 51 to the Defense Waste Processing Facility (DWPF) feed tank (Tank 40). The current WSE practice is to prepare sludge batches in Tank 51 by transferring sludge from other tanks. Discharges of nuclear materials from H Canyon are often added to Tank 51 during sludge batch preparation. The sludge is washed and transferred to Tank 40, the current DWPF feed tank. Prior to transfer of Tank 51 to Tank 40, SRNL typically simulates the Tank Farm and DWPF processes with a Tank 51 sample (referred to as the qualification sample). With the tight schedule constraints for SB7b and the potential need for caustic addition to allow for an acceptable glass processing window, the qualification for SB7b was approached differently than past batches. For SB7b, SRNL prepared a Tank 51 and a Tank 40 sample for qualification. SRNL did not receive the qualification sample from Tank 51 nor did it simulate all of the Tank Farm washing and decanting operations. Instead, SRNL prepared a Tank 51 SB7b sample from samples of Tank 7 and Tank 51, along with a wash solution to adjust the supernatant composition to the final SB7b Tank 51 Tank Farm projections. SRNL then prepared a sample to represent SB7b in Tank 40 by combining portions of the SRNL-prepared Tank 51 SB7b sample and a Tank 40 Sludge Batch 7a (SB7a) sample. The blended sample was 71% Tank 40 (SB7a) and 29% Tank 7/Tank 51 on an insoluble solids basis. This sample is referred to as the SB7b Qualification Sample. The blend represented the highest projected Tank 40 heel (as of May 25, 2011), and thus, the highest

  20. TANK 40 FINAL SB7B CHEMICAL CHARACTERIZATION RESULTS

    SciTech Connect

    Bannochie, C.

    2012-03-15

    /free OH{sup -}/other base, total inorganic carbon/total organic carbon (TIC/TOC) analyses, and Cs-137 gamma scan. Weighted dilutions of slurry were submitted for IC, TIC/TOC, and total base/free OH{sup -}/other base analyses. Activities for U-233, U-235, and Pu-239 were determined from the ICP-MS data for the aqua regia digestions of the Tank 40 WAPS slurry using the specific activity of each isotope. The Pu-241 value was determined from a Pu-238/-241 method developed by SRNL AD and previously described. The following conclusions were drawn from the analytical results reported here: (1) The ratios of the major elements for the SB7b WAPS sample are different from those measured for the SB7a WAPS sample. There is less Al and Mn relative to Fe than the previous sludge batch. (2) The elemental composition of this sample and the analyses conducted here are reasonable and consistent with DWPF batch data measurements in light of DWPF pre-sample concentration and SRAT product heel contributions to the DWPF SRAT receipt analyses. The element ratios for Al/Fe, Ca/Fe, Mn/Fe, and U/Fe agree within 10% between this work and the DWPF Sludge Receipt and Adjustment Tank (SRAT) receipt analyses. (3) Sulfur in the SB7b WAPS sample is 82% soluble, slightly less than results reported for SB3, SB4, and SB6 samples but unlike the 50% insoluble sulfur observed in the SB5 WAPS sample. In addition, 23% of the soluble sulfur is not present as sulfate in SB7b. (4) The average activities of the fissile isotopes of interest in the SB7b WAPS sample are (in {mu}Ci/g of total dried solids): 4.22E-02 U-233, 6.12E-04 U-235, 1.08E+01 Pu-239, and 5.09E+01 Pu-241. The full radionuclide composition will be reported in a future document. (5) The fission product noble metal and Ag concentrations appear to have largely peaked in previous DWPF sludge batches, with the exception of Ru, which still shows a slight increase in SB7b.

  1. SULFATE SOLUBILITY LIMIT VERIFICATION FOR DWPF SLUDGE BATCH 7B

    SciTech Connect

    Fox, K.

    2011-10-03

    the targeted values. The results for the SB7b glasses fabricated with Frit 418 showed an apparent trend of increasing sulfate retention with increasing Na{sub 2}O additions to the 5/25/11 sludge projection. This trend appears contradictory to other recent studies of sulfate retention in Defense Waste Processing Facility (DWPF) type glasses. Additional apparent contradictions to this trend were found in the data collected in the present study. Overall, the results for the SB7b sulfate study glasses with Frit 418 and the 5/25/11 projection with Na{sub 2}O additions showed that subtle changes in this complex glass composition impacted the degree of sulfate retention. These results do however provide confidence that a 0.6 wt % sulfate limit in glass is warranted for Frit 418 with the SB7b compositions evaluated in this study. The results for the SB7b glasses fabricated with Frit 702 are consistent with those of the previous SB7a study in that Frit 702 allowed for higher sulfate retention as compared to Frit 418 for the same sludge compositions. It is recommended that the DWPF implement a sulfate concentration limit of 0.6 wt % in glass for SB7b processing with Frit 418. If a higher than projected sulfate concentration is measured when SB7b processing begins (i.e., if a sulfate concentration higher than 0.6 wt % becomes necessary to achieve targeted waste loadings), DWPF should consider a transition to Frit 702. The sulfate limit could likely be raised to 0.8 wt % by transitioning to this frit. However, if DWPF considers transitioning from Frit 418 to Frit 702, additional glasses should be fabricated to confirm this higher limit due to the issues with incorrect B{sub 2}O{sub 3} concentrations for some of the glasses made with Frit 702 in this study. There are several factors other than sulfate retention that must also be carefully considered prior to changing frit compositions.

  2. CoRoT-7b: SUPER-EARTH OR SUPER-Io?

    SciTech Connect

    Barnes, Rory; Kaib, Nathan A.; Raymond, Sean N.; Greenberg, Richard; Jackson, Brian

    2010-02-01

    CoRoT-7b, a planet about 70% larger than the Earth orbiting a Sun-like star, is the first-discovered rocky exoplanet, and hence has been dubbed a 'super-Earth'. Some initial studies suggested that since the planet is so close to its host star, it receives enough insolation to partially melt its surface. However, these past studies failed to take into consideration the role that tides may play in this system. Even if the planet's eccentricity has always been zero, we show that tidal decay of the semimajor axis could have been large enough that the planet formed on a wider orbit which received less insolation. Moreover, CoRoT-7b could be tidally heated at a rate that dominates its geophysics and drives extreme volcanism. In this case, CoRoT-7b is a 'super-Io' that, like Jupiter's volcanic moon, is dominated by volcanism and rapid resurfacing. Such heating could occur with an eccentricity of just 10{sup -5}. This small value could be driven by CoRoT-7c if its own eccentricity is larger than {approx}10{sup -4}. CoRoT-7b may be the first of a class of planetary super-Ios likely to be revealed by the CoRoT and Kepler spacecraft.

  3. Determination Of Reportable Radionuclides For DWPF Sludge Batch 7B (Macrobatch 9)

    SciTech Connect

    Crawford, C. L.; Diprete, D. P.

    2012-12-17

    The DWPF is receiving radioactive sludge slurry from HLW Tank 40. The radioactive sludge slurry in Tank 40 is a blend of the heel from Sludge Batch 7a (SB7a) and Sludge Batch 7b (SB7b) that was transferred to Tank 40 from Tank 51. The blend of sludge in Tank 40 is also referred to as Macrobatch 9 (MB9). This report develops the list of reportable radionuclides and associated activities as a function of time. Twenty-seven radionuclides have been identified as reportable for DWPF SB7b. Each of these radionuclides has a half-life greater than ten years and contributes more than 0.01% of the radioactivity on a Curie basis at some point from production through the 1100 year period between 2015 and 3115. For SB7b, all of the radionuclides in the Design Basis glass are reportable except for three radionuclides: Pd-107, Cs-135, and Th-230. At no time during the 1100- year period between 2015 and 3115 did any of these three radionuclides contribute to more than 0.01% of the radioactivity on a Curie basis. The radionuclide measurements made for SB7b are the most extensive conducted to date. Some method development/refinement occurred during the conduct of these measurements, leading to lower detection limits and more accurate measurement of some isotopes than was previously possible.

  4. DWPF SIMULANT CPC STUDIES FOR SB7B

    SciTech Connect

    Koopman, D.

    2011-11-01

    same acid stoichiometric factor. The slow acid addition in MCU seemed to alter the reactions that consumed the small excess acid present such that hydrogen generation was promoted relative to sludge-only processing. The coupled test reached higher wt.% total solids, and this likely contributed to the SME cycle hydrogen limit being exceeded at 110% KMA. It is clear from the trends in the SME processing GC data, however, that the frit slurry formic acid contributed to driving the hydrogen generation rate above the SME cycle limit. Hydrogen generation rates after the second frit addition generally exceeded those after the first frit addition. SRAT formate loss increased with increasing acid stoichiometry (15% to 35%). A substantial nitrate gain which was observed to have occurred after acid addition (and nitrite destruction) was reversed to a net nitrate loss in runs with higher acid stoichiometry (nitrate in SRAT product less than sum of sludge nitrate and added nitric acid). Increased ammonium ion formation was also indicated in the runs with nitrate loss. Oxalate loss on the order 20% was indicated in three of the four acid stoichiometry runs and in the coupled flowsheet run. The minimum acid stoichiometry run had no indicated loss. The losses were of the same order as the official analytical uncertainty of the oxalate concentration measurement, but were not randomly distributed about zero loss, so some actual loss was likely occurring. Based on the entire set of SB7b test data, it is recommended that DWPF avoid concentrating additional sludge solids in single SRAT batches to limit the concentrations of noble metals to SB7a processing levels (on a grams noble metal per SRAT batch basis). It is also recommended that DWPF drop the formic acid addition that accompanies the process frit 418 additions, since SME cycle data showed considerable catalytic activity for hydrogen generation from this additional acid (about 5% increase in stoichiometry occurred from the frit

  5. 7. B & O RAILROAD BRIDGE. PHILADELPHIA, PHILADELPHIA CO., PA. ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    7. B & O RAILROAD BRIDGE. PHILADELPHIA, PHILADELPHIA CO., PA. Sec. 1101, MP 3.11. - Northeast Railroad Corridor, Amtrak route between Delaware-Pennsylvania & Pennsylvania-New Jersey state lines, Philadelphia, Philadelphia County, PA

  6. Overview of Materials International Space Station Experiment 7B

    NASA Technical Reports Server (NTRS)

    Jaworske, Donald A.; Siamidis, John

    2009-01-01

    Materials International Space Station Experiment 7B (MISSE 7B) is the most recent in a series of experiments flown on the exterior of International Space Station for the purpose of determining the durability of materials and components in the space environment. A collaborative effort among the Department of Defense, the National Aeronautics and Space Administration, industry, and academia, MISSE 7B will be flying a number of NASA experiments designed to gain knowledge in the area of space environmental effects to mitigate risk for exploration missions. Consisting of trays called Passive Experiment Containers, the suitcase sized payload opens on hinges and allows active and passive experiments contained within to be exposed to the ram and wake or zenith and nadir directions in low Earth orbit, in essence, providing a test bed for atomic oxygen exposure, ultraviolet radiation exposure, charged particle radiation exposure, and thermal cycling. New for MISSE 7B is the ability to monitor experiments actively, with data sent back to Earth via International Space Station communications. NASA?s active and passive experiments cover a range of interest for the Agency. Materials relevant to the Constellation Program include: solar array materials, seal materials, and thermal protection system materials. Materials relevant to the Exploration Technology Development Program include: fabrics for spacesuits, materials for lunar dust mitigation, and new thermal control coatings. Sensors and components on MISSE 7B include: atomic oxygen fluence monitors, ultraviolet radiation sensors, and electro-optical components. In addition, fundamental space environmental durability science experiments are being flown to gather atomic oxygen erosion data and thin film polymer mechanical and optical property data relevant to lunar lander insulation and the James Web Space Telescope. This paper will present an overview of the NASA experiments to be flown on MISSE 7B, along with a summary of the

  7. Neuroendocrine secretory protein 7B2: structure, expression and functions.

    PubMed Central

    Mbikay, M; Seidah, N G; Chrétien, M

    2001-01-01

    7B2 is an acidic protein residing in the secretory granules of neuroendocrine cells. Its sequence has been elucidated in many phyla and species. It shows high similarity among mammals. A Pro-Pro-Asn-Pro-Cys-Pro polyproline motif is its most conserved feature, being carried by both vertebrate and invertebrate sequences. It is biosynthesized as a precursor protein that is cleaved into an N-terminal fragment and a C-terminal peptide. In neuroendocrine cells, 7B2 functions as a specific chaperone for the proprotein convertase (PC) 2. Through the sequence around its Pro-Pro-Asn-Pro-Cys-Pro motif, it binds to an inactive proPC2 and facilitates its transport from the endoplasmic reticulum to later compartments of the secretory pathway where the zymogen is proteolytically matured and activated. Its C-terminal peptide can inhibit PC2 in vitro and may contribute to keep the enzyme transiently inactive in vivo. The PC2-7B2 model defines a new neuroendocrine paradigm whereby proteolytic activation of prohormones and proneuropeptides in the secretory pathway is spatially and temporally regulated by the dynamics of interactions between converting enzymes and their binding proteins. Interestingly, unlike PC2-null mice, which are viable, 7B2-null mutants die early in life from Cushing's disease due to corticotropin ('ACTH') hypersecretion by the neurointermediate lobe, suggesting a possible involvement of 7B2 in secretory granule formation and in secretion regulation. The mechanism of this regulation is yet to be elucidated. 7B2 has been shown to be a good marker of several neuroendocrine cell dysfunctions in humans. The possibility that anomalies in its structure and expression could be aetiological causes of some of these dysfunctions warrants investigation. PMID:11439082

  8. Wnt7b is an important intrinsic regulator of hair follicle stem cell homeostasis and hair follicle cycling.

    PubMed

    Kandyba, Eve; Kobielak, Krzysztof

    2014-04-01

    The hair follicle (HF) is an exceptional mini-organ to study the mechanisms which regulate HF morphogenesis, cycling, hair follicle stem cell (hfSCs) homeostasis, and progeny differentiation. During morphogenesis, Wnt signaling is well-characterized in the initiation of HF patterning but less is known about which particular Wnt ligands are required and whether individual Wnt ligands act in an indispensable or redundant manner during postnatal hfSCs anagen onset and HF cycle progression. Previously, we described the function of the bone morphogenetic protein (BMP) signaling target gene WNT7a in intrinsic regulation of hfSCs homeostasis in vivo. Here, we investigated the role of Wnt7b, which was also intrinsically upregulated in hfSCs during physiological and precocious anagen after BMP inhibition in vivo. We demonstrated Wnt7b to be a direct target of canonical BMP signaling in hfSCs and using Wnt7b conditional gene targeting during HF morphogenesis revealed disrupted HF cycling including a shorter anagen, premature catagen onset with overall shorter hair production, and diminished HF differentiation marker expression. Additionally, we observed that postnatal ablation of Wnt7b resulted in delayed HF activation, affecting both the hair germ and bulge hfSCs but still maintaining a two-step sequence of HF stimulation. Interestingly, Wnt7b cKO hfSCs participated in reformation of the new HF bulge, but with slower self-renewal. These findings demonstrate the importance of intrinsic Wnt7b expression in hfSCs regulation and normal HF cycling and surprisingly reveal a nonredundant role for Wnt7b in the control of HF anagen length and catagen entry which was not compensated by other Wnt ligands. PMID:24222445

  9. Wnt7b is an important intrinsic regulator of hair follicle stem cell homeostasis and hair follicle cycling.

    PubMed

    Kandyba, Eve; Kobielak, Krzysztof

    2014-04-01

    The hair follicle (HF) is an exceptional mini-organ to study the mechanisms which regulate HF morphogenesis, cycling, hair follicle stem cell (hfSCs) homeostasis, and progeny differentiation. During morphogenesis, Wnt signaling is well-characterized in the initiation of HF patterning but less is known about which particular Wnt ligands are required and whether individual Wnt ligands act in an indispensable or redundant manner during postnatal hfSCs anagen onset and HF cycle progression. Previously, we described the function of the bone morphogenetic protein (BMP) signaling target gene WNT7a in intrinsic regulation of hfSCs homeostasis in vivo. Here, we investigated the role of Wnt7b, which was also intrinsically upregulated in hfSCs during physiological and precocious anagen after BMP inhibition in vivo. We demonstrated Wnt7b to be a direct target of canonical BMP signaling in hfSCs and using Wnt7b conditional gene targeting during HF morphogenesis revealed disrupted HF cycling including a shorter anagen, premature catagen onset with overall shorter hair production, and diminished HF differentiation marker expression. Additionally, we observed that postnatal ablation of Wnt7b resulted in delayed HF activation, affecting both the hair germ and bulge hfSCs but still maintaining a two-step sequence of HF stimulation. Interestingly, Wnt7b cKO hfSCs participated in reformation of the new HF bulge, but with slower self-renewal. These findings demonstrate the importance of intrinsic Wnt7b expression in hfSCs regulation and normal HF cycling and surprisingly reveal a nonredundant role for Wnt7b in the control of HF anagen length and catagen entry which was not compensated by other Wnt ligands.

  10. Decreased Lin7b Expression in Layer 5 Pyramidal Neurons May Contribute to Impaired Corticostriatal Connectivity in Huntington Disease

    PubMed Central

    Zucker, Birgit; Kama, Jibrin A.; Kuhn, Alexandre; Thu, Doris; Orlando, Lianna R.; Dunah, Anthone W.; Gokce, Ozgun; Taylor, David M.; Lambeck, Johann; Friedrich, Bernd; Lindenberg, Katrin S.; Faull, Richard L.M.; Weiller, Cornelius; Young, Anne B.; Luthi-Carter, Ruth

    2010-01-01

    Motor dysfunction, cognitive impairment and regional cortical atrophy indicate cerebral cortical involvement in Huntington disease (HD). To address the hypothesis that abnormal corticostriatal connectivity arises from polyglutamine-related alterations in cortical gene expression, we isolated layer 5 cortical neurons by laser-capture microdissection and analyzed transcriptome-wide mRNA changes in them. Enrichment of transcription factor mRNAs including foxp2, tbr1, and neuroD6, and neurotransmission- and plasticity-related RNAs including sema5A, pclo, ntrk2, cntn1 and lin7b were observed. Layer 5 motor cortex neurons of transgenic R6/2 HD mice also demonstrated numerous transcriptomic changes, including decreased expression of mRNAs encoding the lin7 homolog b, (lin7b, also known as veli-2 and mals2). Decreases in LIN7B and CNTN1 RNAs were also detected in human HD layer 5 motor cortex neurons. lin7b, a scaffold protein implicated in synaptic plasticity, neurite outgrowth and cellular polarity, was decreased at the protein level in layer 5 cortical neurons in R6/2 mice and human HD brains. Decreases in Lin7b and Lin7a mRNAs were detected in R6/2 cortex as early as 6 weeks of age, suggesting that this is an early pathogenetic event. Thus, decreased cortical LIN7 expression may contribute to abnormal corticostriatal connectivity in HD. PMID:20720508

  11. THE MASS OF CoRoT-7b

    SciTech Connect

    Hatzes, Artie P.; Wuchterl, Guenther; Fridlund, Malcolm; Gandolfi, Davide; Nachmani, Gil; Mazeh, Tsevi; Valencia, Diana; Hebrard, Guillaume; Borde, Pascal; Carone, Ludmila; Paetzold, Martin; Udry, Stephane; Bouchy, Francois; Deleuil, Magali; Moutou, Claire; Barge, Pierre; Deeg, Hans; Tingley, Brandon; Dvorak, Rudolf; Ferraz-Mello, Sylvio E-mail: malcolm.fridlund@esa.int; and others

    2011-12-10

    The mass of CoRoT-7b, the first transiting super-Earth exoplanet, is still a subject of debate. A wide range of masses have been reported in the literature ranging from as high as 8 M{sub Circled-Plus} to as low as 2.3 M{sub Circled-Plus }. This range in mass is largely due to the activity level of the star that contributes a significant amount of radial velocity (RV) 'jitter' and how the various methods correct this jitter. Although most mass determinations give a density consistent with a rocky planet, the lower value permits a bulk composition that can be up to 50% water. We present an analysis of the CoRoT-7b RV measurements that uses very few and simple assumptions in treating the activity signal. By analyzing those RV data for which multiple measurements were made in a given night, we remove the activity related RV contribution without any a priori model. We argue that the contribution of activity to the final RV curve is negligible and that the K-amplitude due to the planet is well constrained. This yields a mass of 7.42 {+-} 1.21 M{sub Circled-Plus} and a mean density of {rho} = 10.4 {+-} 1.8 gm cm{sup -3}. CoRoT-7b is similar in mass and radius to the second rocky planet to be discovered, Kepler-10b, and within the errors they have identical bulk densities-they are virtual twins. These bulk densities lie close to the density-radius relationship for terrestrial planets similar to what is seen for Mercury. CoRoT-7b and Kepler-10b may have an internal structure more like Mercury than the Earth.

  12. Tank 40 Final SB7b Chemical Characterization Results

    SciTech Connect

    Bannochie, C. J.

    2012-11-06

    A sample of Sludge Batch 7b (SB7b) was taken from Tank 40 in order to obtain radionuclide inventory analyses necessary for compliance with the Waste Acceptance Product Specifications (WAPS). The SB7b WAPS sample was also analyzed for chemical composition including noble metals and fissile constituents. At the Savannah River National Laboratory (SRNL) the 3-L Tank 40 SB7b sample was transferred from the shipping container into a 4-L high density polyethylene bottle and solids were allowed to settle over the weekend. Supernate was then siphoned off and circulated through the shipping container to complete the transfer of the sample. Following thorough mixing of the 3-L sample, a 558 g sub-sample was removed. This sub-sample was then utilized for all subsequent analytical samples. Eight separate aliquots of the slurry were digested, four with HNO{sub 3}/HCl (aqua regia) in sealed Teflon vessels and four with NaOH/Na{sub 2}O{sub 2} (alkali or peroxide fusion) using Zr crucibles. Two Analytical Reference Glass ? 1 (ARG-1) standards were digested along with a blank for each preparation. Each aqua regia digestion and blank was diluted to 1:100 mL with deionized water and submitted to Analytical Development (AD) for inductively coupled plasma ? atomic emission spectroscopy (ICP-AES) analysis, inductively coupled plasma ? mass spectrometry (ICP-MS) analysis, atomic absorption spectroscopy (AA) for As and Se, and cold vapor atomic absorption spectroscopy (CV-AA) for Hg. Equivalent dilutions of the alkali fusion digestions and blank were submitted to AD for ICP-AES analysis. Tank 40 SB7b supernate was collected from a mixed slurry sample in the SRNL Shielded Cells and submitted to AD for ICP-AES, ion chromatography (IC), total base/free OH{sup -}/other base, total inorganic carbon/total organic carbon (TIC/TOC) analyses, and Cs-137 gamma scan. Weighted dilutions of slurry were submitted for IC, TIC/TOC, and total base/free OH-/other base analyses. Activities for U-233, U-235

  13. Thermoelasticity of Fe7C3 under inner core conditions

    NASA Astrophysics Data System (ADS)

    Li, Y.; Vočadlo, L.; Brodholt, J.; Wood, I. G.

    2016-08-01

    It has recently been reported, on the basis of extrapolated experimental data, that the iron carbide, Fe7C3, has shear wave velocities and a Poisson's ratio consistent with the seismological values for the Earth's inner core and thus that Fe7C3 is a strong candidate for the inner core composition. In this study, using ab initio molecular dynamics simulations, we report the thermoelastic properties of Fe7C3 at 350 GPa up to its melting temperature. Due to significant elastic softening prior to melting, the calculated elastic properties, including wave velocities, do indeed agree well with those from seismology. However, the density was found to be much too low (by ~8%) when compared to geophysical data, and therefore, Fe7C3 must be ruled out as a major component of the Earth's inner core.

  14. Characterization of FRA7B, a human common fragile site mapped at the 7p chromosome terminal region.

    PubMed

    Bosco, Nazario; Pelliccia, Franca; Rocchi, Angela

    2010-10-01

    Common fragile sites (CFS) are specific regions of the mammalian chromosomes that are particularly prone to gaps and breaks. They are a cause of genome instability, and the location of many CFS correlates with breakpoints of aberrations recurrent in some cancers. The molecular characterization of some CFS has not clarified the causes of their fragility. In this work, by using fluorescence in situ hybridization analysis with BAC and PAC clones, we determined the DNA sequence of the CFS FRA7B. The FRA7B sequence was then analyzed to identify coding sequences and some structural features possibly involved in fragility. FRA7B spans about 12.2 megabases, and is therefore one of the largest CFS analyzed. It maps at the 7p21.3-22.3 chromosome bands, therefore at the interface of G- and R-band regions that are probably difficult to replicate. A 90-kilobase long sequence that presents very high flexibility values was identified at the very beginning of the more fragile CFS region. Three large genes (THSD7A, SDK1, and MAD1L1) and two miRNA genes (MIRN589 and MIRN339) map in the fragile region. The chromosome band 7p22 is a recurrent breakpoint in chromosome abnormalities in different types of neoplasm. FRA7B is the first characterized CFS located in a chromosome terminal region.

  15. KEPLER-7b: A TRANSITING PLANET WITH UNUSUALLY LOW DENSITY

    SciTech Connect

    Latham, David W.; Buchhave, Lars A.; Furesz, Gabor; Geary, John C.; Borucki, William J.; Koch, David G.; Lissauer, Jack J.; Rowe, Jason F.; Brown, Timothy M.; Basri, Gibor; Batalha, Natalie M.; Caldwell, Douglas A.; Jenkins, Jon M.; Cochran, William D.; Dunham, Edward W.; Gautier, Thomas N.; Howell, Steve B.; Marcy, Geoffrey W.; Monet, David G.

    2010-04-20

    We report on the discovery and confirmation of Kepler-7b, a transiting planet with unusually low density. The mass is less than half that of Jupiter, M {sub P} = 0.43 M {sub J}, but the radius is 50% larger, R {sub P} = 1.48 R {sub J}. The resulting density, {rho}{sub P} = 0.17 g cm{sup -3}, is the second lowest reported so far for an extrasolar planet. The orbital period is fairly long, P = 4.886 days, and the host star is not much hotter than the Sun, T {sub eff} = 6000 K. However, it is more massive and considerably larger than the Sun, M {sub *} = 1.35 M {sub sun} and R {sub *} = 1.84 R {sub sun}, and must be near the end of its life on the main sequence.

  16. DETERMINATION OF REPORTABLE RADIONUCLIDES FOR DWPF SLUDGE BATCH 7B (MACROBATCH 9)

    SciTech Connect

    Crawford, C. L.; Diprete, D. P.

    2014-05-01

    The Waste Acceptance Product Specifications (WAPS) 1.2 require that “The Producer shall report the inventory of radionuclides (in Curies) that have half-lives longer than 10 years and that are, or will be, present in concentrations greater than 0.05 percent of the total inventory for each waste type indexed to the years 2015 and 3115”. As part of the strategy to comply with WAPS 1.2, the Defense Waste Processing Facility (DWPF) will report for each waste type, all radionuclides (with half-lives greater than 10 years) that have concentrations greater than 0.01 percent of the total inventory from time of production through the 1100 year period from 2015 through 3115. The initial listing of radionuclides to be included is based on the design-basis glass as identified in the Waste Form Compliance Plan (WCP) and Waste Form Qualification Report (WQR). However, it is required that this list be expanded if other radionuclides with half-lives greater than 10 years are identified that may meet the greater than 0.01% criterion for Curie content. Specification 1.6 of the WAPS, International Atomic Energy Agency (IAEA) Safeguards Reporting for High Level Waste (HLW), requires that the ratio by weights of the following uranium and plutonium isotopes be reported: U-233, U-234, U-235, U-236, U-238, Pu-238, Pu-239, Pu-240, Pu-241, and Pu- 242. Therefore, the complete set of reportable radionuclides must also include this set of U and Pu isotopes. The DWPF is receiving radioactive sludge slurry from HLW Tank 40. The radioactive sludge slurry in Tank 40 is a blend of the heel from Sludge Batch 7a (SB7a) and Sludge Batch 7b (SB7b) that was transferred to Tank 40 from Tank 51. The blend of sludge in Tank 40 is also referred to as Macrobatch 9 (MB9). This report develops the list of reportable radionuclides and associated activities as a function of time. The DWPF will use this list and the activities as one of the inputs for the development of the Production Records that relate to

  17. Determination Of Reportable Radionuclides For DWPF Sludge Batch 7B (Macrobatch 9)

    SciTech Connect

    Crawford, C. L.; DiPrete, D. P.

    2013-08-22

    The Waste Acceptance Product Specifications (WAPS) 1.2 require that “The Producer shall report the inventory of radionuclides (in Curies) that have half-lives longer than 10 years and that are, or will be, present in concentrations greater than 0.05 percent of the total inventory for each waste type indexed to the years 2015 and 3115”. As part of the strategy to comply with WAPS 1.2, the Defense Waste Processing Facility (DWPF) will report for each waste type, all radionuclides (with half-lives greater than 10 years) that have concentrations greater than 0.01 percent of the total inventory from time of production through the 1100 year period from 2015 through 3115. The initial listing of radionuclides to be included is based on the design-basis glass as identified in the Waste Form Compliance Plan (WCP) and Waste Form Qualification Report (WQR). However, it is required that this list be expanded if other radionuclides with half-lives greater than 10 years are identified that may meet the greater than 0.01% criterion for Curie content. Specification 1.6 of the WAPS, International Atomic Energy Agency (IAEA) Safeguards Reporting for High Level Waste (HLW), requires that the ratio by weights of the following uranium and plutonium isotopes be reported: U-233, U-234, U-235, U-236, U-238, Pu-238, Pu-239, Pu-240, Pu-241, and Pu-242. Therefore, the complete set of reportable radionuclides must also include this set of U and Pu isotopes. The DWPF is receiving radioactive sludge slurry from HLW Tank 40. The radioactive sludge slurry in Tank 40 is a blend of the heel from Sludge Batch 7a (SB7a) and Sludge Batch 7b (SB7b) that was transferred to Tank 40 from Tank 51. The blend of sludge in Tank 40 is also referred to as Macrobatch 9 (MB9). This report develops the list of reportable radionuclides and associated activities as a function of time. The DWPF will use this list and the activities as one of the inputs for the development of the Production Records that relate to

  18. Critical roles for the COOH terminus of the Cu-ATPase ATP7B in protein stability, trans-Golgi network retention, copper sensing, and retrograde trafficking.

    PubMed

    Braiterman, L; Nyasae, L; Leves, F; Hubbard, A L

    2011-07-01

    ATP7A and ATP7B are copper-transporting P-type ATPases that are essential to eukaryotic copper homeostasis and must traffic between intracellular compartments to carry out their functions. Previously, we identified a nine-amino acid sequence (F37-E45) in the NH(2) terminus of ATP7B that is required to retain the protein in the Golgi when copper levels are low and target it apically in polarized hepatic cells when copper levels rise. To understand further the mechanisms regulating the intracellular dynamics of ATP7B, using multiple functional assays, we characterized the protein phenotypes of 10 engineered and Wilson disease-associated mutations in the ATP7B COOH terminus in polarized hepatic cells and fibroblasts. We also examined the behavior of a chimera between ATP7B and ATP7A. Our results clearly demonstrate the importance of the COOH terminus of ATP7B in the protein's copper-responsive apical trafficking. L1373 at the end of transmembrane domain 8 is required for protein stability and Golgi retention in low copper, the trileucine motif (L1454-L1456) is required for retrograde trafficking, and the COOH terminus of ATP7B exhibits a higher sensitivity to copper than does ATP7A. Importantly, our results demonstrating that four Wilson disease-associated missense mutations behaved in a wild-type manner in all our assays, together with current information in the literature, raise the possibility that several may not be disease-causing mutations.

  19. EOS7C-ECBM Version 1.0

    2012-11-14

    EOS7C is an equation of state module for the TOUGH2 program for CO2 or N2 in Methane (CH4) Reservoirs. In the present work, additions have been made to the EOS7C Version 1.0 module to include the Enhanced Coal Bed Methane (ECBM) modifications developed by Webb (2003). In addition, the Dusty Gas Model for gas-phase diffusion (Webb 2001) has been included. The ECBM modification to the EOS7C equation of state incorporate the extended Langmuir isothem formore » sorbing gases, including the change in porosity associated with the sorbed gas mass. Comparison to hand calculations for pure gas and binary mixture shows very good agreement. Application to a CO2 well injection problem by Law et al. (2002). The Dusty Model modification add options to calculate gas diffusion using the Dusty-Gas Model including separate and coupled approaches. Comparison to low-permeability pure gas diffusion data shows excellent agreement. The results from the DGM are compared to the Fick's Law behavior for diffusion across a capillary fringe. The differences between the models are small due to the relatively high permeability (10-11 m2) of the problem.« less

  20. EOS7C-ECBM Version 1.0

    SciTech Connect

    2012-11-14

    EOS7C is an equation of state module for the TOUGH2 program for CO2 or N2 in Methane (CH4) Reservoirs. In the present work, additions have been made to the EOS7C Version 1.0 module to include the Enhanced Coal Bed Methane (ECBM) modifications developed by Webb (2003). In addition, the Dusty Gas Model for gas-phase diffusion (Webb 2001) has been included. The ECBM modification to the EOS7C equation of state incorporate the extended Langmuir isothem for sorbing gases, including the change in porosity associated with the sorbed gas mass. Comparison to hand calculations for pure gas and binary mixture shows very good agreement. Application to a CO2 well injection problem by Law et al. (2002). The Dusty Model modification add options to calculate gas diffusion using the Dusty-Gas Model including separate and coupled approaches. Comparison to low-permeability pure gas diffusion data shows excellent agreement. The results from the DGM are compared to the Fick's Law behavior for diffusion across a capillary fringe. The differences between the models are small due to the relatively high permeability (10-11 m2) of the problem.

  1. Defective roles of ATP7B missense mutations in cellular copper tolerance and copper excretion.

    PubMed

    Zhu, Min; Dong, Yi; Ni, Wang; Wu, Zhi-Ying

    2015-07-01

    Wilson's disease (WD) is a hereditary disorder of copper metabolism resulting from mutations within ATP7B. Clinical investigations showed that ATP7B missense mutations cause a wide variety of symptoms in WD patients, which implies that those mutations might affect ATP7B function in a number of ways and each would have deleterious consequences on normal copper distribution and lead to WD. Nonetheless, it is still unknown about the influences of those mutations on ATP7B function of increasing copper excretion and enhancing cellular copper tolerance. Here we established the stable expression cell lines of wild-type (WT) ATP7B and its four missense mutants (R778L, R919G, T935M and P992L), tested cellular copper tolerance and copper excretion using those cell lines, and also observed cellular distribution of WT ATP7B proteins and those mutants in transiently transfected cells. We found that extrinsic expressing WT ATP7B reduced CuCl2-induced copper accumulation and enhanced cellular copper tolerance by accelerating copper excretion, which was selectively compromised by R778L and P992L mutations. Further investigation showed that R778L mutation disrupted the subcellular localization and trafficking of ATP7B proteins, whereas P992L mutation only affected the trafficking of ATP7B. This indicates that ATP7B missense mutants have distinct effects on cellular copper tolerance.

  2. The neuroendocrine polypeptide 7B2 is an endogenous inhibitor of prohormone convertase PC2.

    PubMed Central

    Martens, G J; Braks, J A; Eib, D W; Zhou, Y; Lindberg, I

    1994-01-01

    The subtilisin-like prohormone convertase PC2 and the polypeptide 7B2 (an intracellularly cleaved protein of unknown function) are both selectively present in the regulated secretory pathway of neurons and endocrine cells. Here we demonstrate that intact recombinant 7B2 is a potent inhibitor of PC2 and prevents proPC2 cleavage in vitro, whereas the 7B2 cleavage product is virtually inactive. The PC2-related proteinase PC1/PC3 is not inhibited by 7B2. Furthermore, the carboxyl-terminal half of the 7B2 protein sequence is distantly related to the so-called potato inhibitor I family (which includes subtilisin inhibitors). Our findings indicate that 7B2 is a physiological inhibitor of PC2 and may provide alternative avenues for the manipulation of peptide hormone levels. Images PMID:8016065

  3. Novel alternative splice variants of rat phosphodiesterase 7B showing unique tissue-specific expression and phosphorylation.

    PubMed Central

    Sasaki, Takashi; Kotera, Jun; Omori, Kenji

    2002-01-01

    cDNA species coding for novel variants of cyclic-AMP-specific phosphodiesterases (PDEs), namely the PDE7B family, were isolated from rats and characterized. Rat PDE7B1 (RNPDE7B1) was composed of 446 amino acid residues. Rat PDE7B2 (RNPDE7B2) and PDE7B3 (RNPDE7B3), which possessed unique N-terminal sequences, consisted of 359 and 459 residues respectively. Northern hybridization analysis showed that rat PDE7B transcripts were particularly abundant in the striatum and testis. PCR analyses revealed that rat PDE7B2 transcripts were restricted to the testis and that low levels of PDE7B3 transcripts were expressed in the heart, lung and skeletal muscle. In situ hybridization analysis demonstrated that rat PDE7B transcripts were expressed in striatal neurons and spermatocytes. In spermatocytes, rat PDE7B transcripts were expressed in a stage-specific manner during spermatogenesis. The K(m) values of recombinant rat PDE7B1, PDE7B2 and PDE7B3 for cAMP were 0.05, 0.07 and 0.05 microM respectively. Each rat PDE7B variant was the most sensitive to 3-isobutyl-1-methylxanthine (IC(50) 1.5-2.1 microM). Two phosphorylation sites for cAMP-dependent protein kinase (PKA) were found in rat PDE7B1 and PDE7B3, whereas rat PDE7B2 possessed one site. PKA-dependent phosphorylation was observed in C-terminal phosphorylation sites of three rat PDE7B variants, in addition to unique N-terminal regions of rat PDE7B1 and PDE7B3. Unique tissue distribution and PKA-dependent phosphorylation of PDE7B variants suggested that each variant has a specific role for cellular functions via cAMP signalling in various tissues. PMID:11772393

  4. Let-7b Inhibits Human Cancer Phenotype by Targeting Cytochrome P450 Epoxygenase 2J2

    PubMed Central

    Yang, Shenglan; Gong, Wei; Wang, Yan; Cianflone, Katherine; Tang, Jiarong; Wang, Dao Wen

    2012-01-01

    Background MicroRNAs (miRNAs) are small, noncoding RNA molecules of 20 to 22 nucleotides that regulate gene expression by binding to their 3′ untranslated region (3′UTR). Increasing data implicate altered miRNA participation in the progress of cancer. We previously reported that CYP2J2 epoxygenase promotes human cancer phenotypes. But whether and how CYP2J2 is regulated by miRNA is not understood. Methods and Results Using bioinformatics analysis, we found potential target sites for miRNA let-7b in 3′UTR of human CYP2J2. Luciferase and western blot assays revealed that CYP2J2 was regulated by let-7b. In addition, let-7b decreased the enzymatic activity of endogenous CYP2J2. Furthermore, let-7b may diminish cell proliferation and promote cell apoptosis of tumor cells via posttranscriptional repression of CYP2J2. Tumor xenografts were induced in nude mice by subcutaneous injection of MDA-MB-435 cells. The let-7b expression vector, pSilencer-let-7b, was injected through tail vein every 3 weeks. Let-7b significantly inhibited the tumor phenotype by targeting CYP2J2. Moreover, quantitative real-time polymerase chain reaction and western blotting were used to determine the expression levels of let-7b and CYP2J2 protein from 18 matched lung squamous cell cancer and adjacent normal lung tissues; the expression level of CYP2J2 was inversely proportional to that of let-7b. Conclusions Our results demonstrated that the decreased expression of let-7b could lead to the high expression of CYP2J2 protein in cancerous tissues. These findings suggest that miRNA let-7b reduces CYP2J2 expression, which may contribute to inhibiting tumor phenotypes. PMID:22761738

  5. Phosphorylation and Alternative Splicing of 7B2 Reduce Prohormone Convertase 2 Activation

    PubMed Central

    Ramos-Molina, Bruno

    2015-01-01

    FAM20C is a secretory kinase responsible for the phosphorylation of multiple secreted proteins in mammalian cells; it has been shown to phosphorylate serine residues within a variety of different bone proteins. In this work we demonstrate that FAM20C also phosphorylates threonines, specifically those within the N-terminal domain of the neuroendocrine chaperone 7B2. Analysis of the primary sequence of 7B2 revealed that three threonine residues in its N-terminal domain are located within FAM20C consensus motifs: Thr73, Thr99, and Thr111. The individual substitution of Thr73 and Thr111 residues by neutral alanines caused a marked decrease in the total phosphorylation of 7B2. Furthermore, the phosphomimetic substitution of Thr111 by Glu clearly diminished the ability of 7B2 to activate pro-prohormone convertase 2 (PC2) in 7B2-lacking SK-N-MC neuroblastoma cells, suggesting that the phosphorylation of this residue critically impacts the 7B2-proPC2 interaction. However, the phosphomimetic mutation did not alter 7B2's ability to function as an antiaggregant for human islet amyloid polypeptide. FAM20C-mediated phosphorylation of a common alternatively spliced variant of human 7B2 that lacks Ala100 (thus eliminating the Thr99 phosphorylation consensus site) was similar to the Ala-containing protein, but this variant did not activate proPC2 as efficiently as the Ala-containing protein. Although threonines within 7B2 were phosphorylated efficiently, FAM20C was incapable of performing the well-known regulatory threonine phosphorylation of the molecular chaperone binding immunoglobulin protein. Taken together, these results indicate that FAM20C plays a role in 7B2-mediated proPC2 activation by phosphorylating residue Thr111; and that 7B2 function is regulated by alternative splicing. PMID:25811241

  6. Copper binding triggers compaction in N-terminal tail of human copper pump ATP7B.

    PubMed

    Mondol, Tanumoy; Åden, Jörgen; Wittung-Stafshede, Pernilla

    2016-02-12

    Protein conformational changes are fundamental to biological reactions. For copper ion transport, the multi-domain protein ATP7B in the Golgi network receives copper from the cytoplasmic copper chaperone Atox1 and, with energy from ATP hydrolysis, moves the metal to the lumen for loading of copper-dependent enzymes. Although anticipated, conformational changes involved in ATP7B's functional cycle remain elusive. Using spectroscopic methods we here demonstrate that the four most N-terminal metal-binding domains in ATP7B, upon stoichiometric copper addition, adopt a more compact arrangement which has a higher thermal stability than in the absence of copper. In contrast to previous reports, no stable complex was found in solution between the metal-binding domains and the nucleotide-binding domain of ATP7B. Metal-dependent movement of the first four metal-binding domains in ATP7B may be a trigger that initiates the overall catalytic cycle.

  7. CYP7B1 Enzyme Deletion Impairs Reproductive Behaviors in Male Mice

    PubMed Central

    Oyola, Mario G.; Zuloaga, Damian G.; Carbone, David; Malysz, Anna M.; Acevedo-Rodriguez, Alexandra; Handa, Robert J.

    2015-01-01

    In addition to androgenic properties mediated via androgen receptors, dihydrotestosterone (DHT) also regulates estrogenic functions via an alternate pathway. These estrogenic functions of DHT are mediated by its metabolite 5α-androstane-3β, 17β-diol (3β-diol) binding to estrogen receptor β (ERβ). CYP7B1 enzyme converts 3β-diol to inactive 6α- or 7α-triols and plays an important role as a regulator of estrogenic functions mediated by 3β-diol. Using a mutant mouse carrying a null mutation for the CYP7B1 gene (CYP7B1KO), we examined the contribution of CYP7B1 on physiology and behavior. Male, gonadectomized (GDX) CYP7B1KO and their wild type (WT) littermates were assessed for their behavioral phenotype, anxiety-related behavioral measures, and hypothalamic pituitary adrenal axis reactivity. No significant effects of genotype were evident in anxiety-like behaviors in open field (OFA), light-dark (L/D) exploration, and elevated plus maze (EPM). T significantly reduced open arm time on the EPM while not affecting L/D exploratory and OFA behaviors in CYP7B1KO and WT littermates. T also attenuated the corticosterone response to EPM in both genotypes. In GDX animals, T was able to reinstate male-specific reproductive behaviors (latencies and number of mounts, intromission, and ejaculations) in the WT but not in the CYP7B1KO mice. The male reproductive behavior defect in CYP7B1KO seems to be due to their inability to distinguish olfactory cues from a behavioral estrus female. CYP7B1KO mice also showed a reduction in androgen receptor mRNA expression in the olfactory bulb. Our findings suggest a novel role for the CYP7B1 enzyme in the regulation of male reproductive behaviors. PMID:25849728

  8. Analysis and application of ATP7B gene mutations in 35 patients with hepatolenticular degeneration.

    PubMed

    Zong, Y N; Kong, X D

    2015-01-01

    We investigated the genetic mutations involved in Wilson's disease to improve prenatal genetic diagnosis and presymptomatic diagnosis. The polymerase chain reaction (PCR) was used to amplify the exons and exon-intron boundaries of the ATP7B gene in 35 Wilson's disease pedigrees. The PCR products were further analyzed by Sanger sequencing. Prenatal genetic diagnoses were performed by chorionic villus sampling after the genotypes of parents of the probands were identified. The overall mutation detection frequency was 92.9%. A total of 24 distinct mutations were detected, seven of which are novel: A1291T (c.3871G>A), c.2593_2594insGTCA, c.2790_2792delCAT, c.3661_3663delGGG, c.3700delG, c.4094_4097delCTGT, and IVS6+1G>A. Three mutations, R778L (c.2333G>T) (45.7%), A874V (c.2621C>T) (7.1%), and P992L (c.2975C>T) (7.1%) are relatively frequent. Two presymptomatic patients were detected through familial screening, and they began taking medicine after diagnosis. Of the subjects with Wilson's disease pedigrees who had received a prenatal genetic diagnosis, three fetuses were normal and one was a carrier. Twenty-four distinct mutations were identified, and our knowledge of the population genetics of Wilson's disease in China has therefore improved. For pedigrees with the Wilson's disease, genetic counseling, prenatal diagnosis, and presymptomatic diagnosis by Sanger sequencing and haplotype analysis are feasible. PMID:26782526

  9. Results of International Space Station Vehicle Materials Exposed on MISSE-7B

    NASA Technical Reports Server (NTRS)

    Finckenor, Miria M.; Golden, Johnny L.; O'Rourke, Mary Jane E.; Kravchenko, Michael

    2012-01-01

    Materials samples were exposed to the low Earth orbit (LEO) environment as part of the MISSE-7B flight experiment for 18 months. Optical properties, thickness/mass loss, surface elemental analysis, visual and microscopic analysis for surface change are some of the techniques employed in this investigation. Where possible, the MISSE-7B results are compared to analyses from other LEO experiments. ISS materials currently flying on MISSE-8 are also discussed.

  10. Genome Sequence of Leucobacter sp. 4J7B1, a Plant-Osmoprotectant Soil Microorganism

    PubMed Central

    Vílchez, J. I.; García-Fontana, C.; Calvo, C.; González-López, J.

    2015-01-01

    We report the first genome sequence for Leucobacter sp. 4J7B1, a newly described desiccation-tolerant strain. The complete genome sequence of Leucobacter sp. 4J7B1 has been sequenced and is estimated to be around 3.5 Mb in size, with an average GC content of 62.18%. We predict 2,953 protein-coding sequences. PMID:25999566

  11. Copper does not alter the intracellular distribution of ATP7B, a copper-transporting ATPase.

    PubMed

    Harada, M; Sakisaka, S; Kawaguchi, T; Kimura, R; Taniguchi, E; Koga, H; Hanada, S; Baba, S; Furuta, K; Kumashiro, R; Sugiyama, T; Sata, M

    2000-09-01

    Wilson's disease is a genetic disorder characterized by the accumulation of copper in the body due to a defect of biliary copper excretion. However, the mechanism of biliary copper excretion has not been fully clarified. We examined the effect of copper on the intracellular localization of the Wilson disease gene product (ATP7B) and green fluorescent protein (GFP)-tagged ATP7B in a human hepatoma cell line (Huh7). The intracellular organelles were visualized by fluorescence microscopy. GFP-ATP7B colocalized with late endosome markers, but not with endoplasmic reticulum, Golgi, or lysosome markers in both the steady and copper-loaded states. ATP7B mainly localized at the perinuclear regions in both states. These results suggest that the main localization of ATP7B is in the late endosomes in both the steady and copper-loaded states. ATP7B seems to translocate copper from the cytosol to the late endosomal lumen, thus participating in biliary copper excretion via lysosomes.

  12. sept7b is required for the differentiation of pancreatic endocrine progenitors

    PubMed Central

    Dash, Surjya Narayan; Hakonen, Elina; Ustinov, Jarkko; Otonkoski, Timo; Andersson, Olov; Lehtonen, Sanna

    2016-01-01

    Protection or restoration of pancreatic β-cell mass as a therapeutic treatment for type 1 diabetes requires understanding of the mechanisms that drive the specification and development of pancreatic endocrine cells. Septins are filamentous small GTPases that function in the regulation of cell division, cytoskeletal organization and membrane remodeling, and are involved in various tissue-specific developmental processes. However, their role in pancreatic endocrine cell differentiation remains unknown. Here we show by functional manipulation techniques in transgenic zebrafish lines that suppression of sept7b, the zebrafish ortholog of human SEPT7, profoundly increases the number of endocrine progenitors but limits their differentiation, leading to reduction in β- and α-cell mass. Furthermore, we discovered that shh (sonic hedgehog) expression in the endoderm, essential for the development of pancreatic progenitors of the dorsal pancreatic bud, is absent in larvae depleted of sept7b. We also discovered that sept7b is important for the differentiation of ventral pancreatic bud-derived cells: sept7b-depleted larvae exhibit downregulation of Notch receptors notch1a and notch1b and show precocious differentiation of NeuroD-positive endocrine cells in the intrapancreatic duct and gut epithelium. Collectively, this study provides a novel insight into the development of pancreatic endocrine progenitors, revealing an essential role for sept7b in endocrine progenitor differentiation. PMID:27114183

  13. Mutations in COX7B cause microphthalmia with linear skin lesions, an unconventional mitochondrial disease.

    PubMed

    Indrieri, Alessia; van Rahden, Vanessa Alexandra; Tiranti, Valeria; Morleo, Manuela; Iaconis, Daniela; Tammaro, Roberta; D'Amato, Ilaria; Conte, Ivan; Maystadt, Isabelle; Demuth, Stephanie; Zvulunov, Alex; Kutsche, Kerstin; Zeviani, Massimo; Franco, Brunella

    2012-11-01

    Microphthalmia with linear skin lesions (MLS) is an X-linked dominant male-lethal disorder associated with mutations in holocytochrome c-type synthase (HCCS), which encodes a crucial player of the mitochondrial respiratory chain (MRC). Unlike other mitochondrial diseases, MLS is characterized by a well-recognizable neurodevelopmental phenotype. Interestingly, not all clinically diagnosed MLS cases have mutations in HCCS, thus suggesting genetic heterogeneity for this disorder. Among the possible candidates, we analyzed the X-linked COX7B and found deleterious de novo mutations in two simplex cases and a nonsense mutation, which segregates with the disease, in a familial case. COX7B encodes a poorly characterized structural subunit of cytochrome c oxidase (COX), the MRC complex IV. We demonstrated that COX7B is indispensable for COX assembly, COX activity, and mitochondrial respiration. Downregulation of the COX7B ortholog (cox7B) in medaka (Oryzias latipes) resulted in microcephaly and microphthalmia that recapitulated the MLS phenotype and demonstrated an essential function of complex IV activity in vertebrate CNS development. Our results indicate an evolutionary conserved role of the MRC complexes III and IV for the proper development of the CNS in vertebrates and uncover a group of mitochondrial diseases hallmarked by a developmental phenotype.

  14. Otud7b facilitates T cell activation and inflammatory responses by regulating Zap70 ubiquitination.

    PubMed

    Hu, Hongbo; Wang, Hui; Xiao, Yichuan; Jin, Jin; Chang, Jae-Hoon; Zou, Qiang; Xie, Xiaoping; Cheng, Xuhong; Sun, Shao-Cong

    2016-03-01

    Signal transduction from the T cell receptor (TCR) is crucial for T cell-mediated immune responses and, when deregulated, also contributes to the development of autoimmunity. How TCR signaling is regulated is incompletely understood. In this study, we demonstrate a ubiquitin-dependent mechanism in which the deubiquitinase Otud7b has a crucial role in facilitating TCR signaling. Upon TCR ligation, Otud7b is rapidly recruited to the tyrosine kinase Zap70, a central mediator of TCR-proximal signaling. Otud7b deficiency attenuates the activation of Zap70 and its downstream pathways and impairs T cell activation and differentiation, rendering mice refractory to T cell-mediated autoimmune and inflammatory responses. Otud7b facilitated Zap70 activation by deubiquitinating Zap70, thus preventing the association of Zap70 with the negative-regulatory phosphatases Sts1 and Sts2. These findings establish Otud7b as a positive regulator of TCR-proximal signaling and T cell activation, highlighting the importance of deubiquitination in regulating Zap70 function. PMID:26903241

  15. Functional characterization of new mutations in Wilson disease gene (ATP7B) using the yeast model.

    PubMed

    Papur, Ozlenen Simsek; Terzioglu, Orhan; Koc, Ahmet

    2015-01-01

    The Wilson disease gene, a copper transporting ATPase (Atp7b), is responsible for the sequestration of Cu into secretory vesicles, and this function is exhibited by the orthologous Ccc2p in the yeast. In this study, we aimed to characterize clinically relevant new mutations of human ATP7B (p.T788I, p.V1036I and p.R1038G-fsX83) in yeast lacking the CCC2 gene. Expression of human wild type ATP7B gene in ccc2Δ mutant yeast restored the growth deficiency and copper transport activity; however, expression of the mutant forms did not restore the copper transport functions and only partially supported the cell growth. Our data support that p.T788I, p.V1036I and p.R1038G-fsX83 mutations cause functional deficiency in ATP7B functions and suggest that these residues are important for normal ATP7B function.

  16. MicroRNA let-7c Is Downregulated in Prostate Cancer and Suppresses Prostate Cancer Growth

    PubMed Central

    Nadiminty, Nagalakshmi; Tummala, Ramakumar; Lou, Wei; Zhu, Yezi; Shi, Xu-Bao; Zou, June X.; Chen, Hongwu; Zhang, Jin; Chen, Xinbin; Luo, Jun; deVere White, Ralph W.; Kung, Hsing-Jien; Evans, Christopher P.; Gao, Allen C.

    2012-01-01

    Purpose Prostate cancer (PCa) is characterized by deregulated expression of several tumor suppressor or oncogenic miRNAs. The objective of this study was the identification and characterization of miR-let-7c as a potential tumor suppressor in PCa. Experimental Design Levels of expression of miR-let-7c were examined in human PCa cell lines and tissues using qRT-PCR and in situ hybridization. Let-7c was overexpressed or suppressed to assess the effects on the growth of human PCa cell lines. Lentiviral-mediated re-expression of let-7c was utilized to assess the effects on human PCa xenografts. Results We identified miR-let-7c as a potential tumor suppressor in PCa. Expression of let-7c is downregulated in castration-resistant prostate cancer (CRPC) cells. Overexpression of let-7c decreased while downregulation of let-7c increased cell proliferation, clonogenicity and anchorage-independent growth of PCa cells in vitro. Suppression of let-7c expression enhanced the ability of androgen-sensitive PCa cells to grow in androgen-deprived conditions in vitro. Reconstitution of Let-7c by lentiviral-mediated intratumoral delivery significantly reduced tumor burden in xenografts of human PCa cells. Furthermore, let-7c expression is downregulated in clinical PCa specimens compared to their matched benign tissues, while the expression of Lin28, a master regulator of let-7 miRNA processing, is upregulated in clinical PCa specimens. Conclusions These results demonstrate that microRNA let-7c is downregulated in PCa and functions as a tumor suppressor, and is a potential therapeutic target for PCa. PMID:22479342

  17. Growth characteristics of primary M7C3 carbide in hypereutectic Fe-Cr-C alloy.

    PubMed

    Liu, Sha; Zhou, Yefei; Xing, Xiaolei; Wang, Jibo; Ren, Xuejun; Yang, Qingxiang

    2016-01-01

    The microstructure of the hypereutectic Fe-Cr-C alloy is observed by optical microscopy (OM). The initial growth morphology, the crystallographic structure, the semi-molten morphology and the stacking faults of the primary M7C3 carbide are observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The in-suit growth process of the primary M7C3 carbide was observed by confocal laser microscope (CLM). It is found that the primary M7C3 carbide in hypereutectic Fe-Cr-C alloy is irregular polygonal shape with several hollows in the center and gaps on the edge. Some primary M7C3 carbides are formed by layers of shell or/and consist of multiple parts. In the initial growth period, the primary M7C3 carbide forms protrusion parallel to {} crystal planes. The extending and revolving protrusion forms the carbide shell. The electron backscattered diffraction (EBSD) maps show that the primary M7C3 carbide consists of multiple parts. The semi-molten M7C3 carbide contains unmelted shell and several small-scale carbides inside, which further proves that the primary M7C3 carbide is not an overall block. It is believed that the coalescence of the primary M7C3 carbides is ascribed to the growing condition of the protrusion and the gap filling process. PMID:27596718

  18. Growth characteristics of primary M7C3 carbide in hypereutectic Fe-Cr-C alloy.

    PubMed

    Liu, Sha; Zhou, Yefei; Xing, Xiaolei; Wang, Jibo; Ren, Xuejun; Yang, Qingxiang

    2016-09-06

    The microstructure of the hypereutectic Fe-Cr-C alloy is observed by optical microscopy (OM). The initial growth morphology, the crystallographic structure, the semi-molten morphology and the stacking faults of the primary M7C3 carbide are observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The in-suit growth process of the primary M7C3 carbide was observed by confocal laser microscope (CLM). It is found that the primary M7C3 carbide in hypereutectic Fe-Cr-C alloy is irregular polygonal shape with several hollows in the center and gaps on the edge. Some primary M7C3 carbides are formed by layers of shell or/and consist of multiple parts. In the initial growth period, the primary M7C3 carbide forms protrusion parallel to {} crystal planes. The extending and revolving protrusion forms the carbide shell. The electron backscattered diffraction (EBSD) maps show that the primary M7C3 carbide consists of multiple parts. The semi-molten M7C3 carbide contains unmelted shell and several small-scale carbides inside, which further proves that the primary M7C3 carbide is not an overall block. It is believed that the coalescence of the primary M7C3 carbides is ascribed to the growing condition of the protrusion and the gap filling process.

  19. Growth characteristics of primary M7C3 carbide in hypereutectic Fe-Cr-C alloy

    NASA Astrophysics Data System (ADS)

    Liu, Sha; Zhou, Yefei; Xing, Xiaolei; Wang, Jibo; Ren, Xuejun; Yang, Qingxiang

    2016-09-01

    The microstructure of the hypereutectic Fe-Cr-C alloy is observed by optical microscopy (OM). The initial growth morphology, the crystallographic structure, the semi-molten morphology and the stacking faults of the primary M7C3 carbide are observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The in-suit growth process of the primary M7C3 carbide was observed by confocal laser microscope (CLM). It is found that the primary M7C3 carbide in hypereutectic Fe-Cr-C alloy is irregular polygonal shape with several hollows in the center and gaps on the edge. Some primary M7C3 carbides are formed by layers of shell or/and consist of multiple parts. In the initial growth period, the primary M7C3 carbide forms protrusion parallel to {} crystal planes. The extending and revolving protrusion forms the carbide shell. The electron backscattered diffraction (EBSD) maps show that the primary M7C3 carbide consists of multiple parts. The semi-molten M7C3 carbide contains unmelted shell and several small-scale carbides inside, which further proves that the primary M7C3 carbide is not an overall block. It is believed that the coalescence of the primary M7C3 carbides is ascribed to the growing condition of the protrusion and the gap filling process.

  20. Growth characteristics of primary M7C3 carbide in hypereutectic Fe-Cr-C alloy

    PubMed Central

    Liu, Sha; Zhou, Yefei; Xing, Xiaolei; Wang, Jibo; Ren, Xuejun; Yang, Qingxiang

    2016-01-01

    The microstructure of the hypereutectic Fe-Cr-C alloy is observed by optical microscopy (OM). The initial growth morphology, the crystallographic structure, the semi-molten morphology and the stacking faults of the primary M7C3 carbide are observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The in-suit growth process of the primary M7C3 carbide was observed by confocal laser microscope (CLM). It is found that the primary M7C3 carbide in hypereutectic Fe-Cr-C alloy is irregular polygonal shape with several hollows in the center and gaps on the edge. Some primary M7C3 carbides are formed by layers of shell or/and consist of multiple parts. In the initial growth period, the primary M7C3 carbide forms protrusion parallel to {} crystal planes. The extending and revolving protrusion forms the carbide shell. The electron backscattered diffraction (EBSD) maps show that the primary M7C3 carbide consists of multiple parts. The semi-molten M7C3 carbide contains unmelted shell and several small-scale carbides inside, which further proves that the primary M7C3 carbide is not an overall block. It is believed that the coalescence of the primary M7C3 carbides is ascribed to the growing condition of the protrusion and the gap filling process. PMID:27596718

  1. Let-7b-mediated suppression of basigin expression and metastasis in mouse melanoma cells

    SciTech Connect

    Fu, Tzu-Yen; Chang, Chia-Che; Lin, Chun-Ting; Lai, Cong-Hao; Peng, Shao-Yu; Ko, Yi-Ju; Tang, Pin-Chi

    2011-02-15

    Basigin (Bsg), also called extracellular matrix metalloproteinase inducer (EMMPRIN), is highly expressed on the surface of tumor cells and stimulates adjacent fibroblasts or tumor cells to produce matrix metalloproteinases (mmps). It has been shown that Bsg plays an important role in growth, development, cell differentiation, and tumor progression. MicroRNAs (miRNAs) are a class of short endogenous non-protein coding RNAs of 20-25 nucleotides (nt) that function as post-transcriptional regulators of gene expression by base-pairing to their target mRNAs and thereby mediate cleavage of target mRNAs or translational repression. In this study, let-7b, one of the let-7 family members, was investigated for its effect on the growth and invasiveness of the mouse melanoma cell line B16-F10. We have shown that let-7b can suppress the expression of Bsg in B16-F10 cells and also provided evidence that this suppression could result in the indirect suppression of mmp-9. The ability of B16-F10 cells transfected with let-7b to invade or migrate was significantly reduced. In addition, let-7b transfected B16-F10 cells displayed an inhibition of both cellular proliferation and colony formation. Furthermore, it was shown that the overexpression of let-7b in B16-F10 cells could reduce lung metastasis. Taken together, the present study identifies let-7b as a tumor suppressor that represses cancer cell proliferation and migration as well as tumor metastasis in mouse melanoma cells.

  2. Site-Directed Mutagenesis, in Vivo Electroporation and Mass Spectrometry in Search for Determinants of the Subcellular Targeting of Rab7b Paralogue in the Model Eukaryote Paramecium Octaurelia

    PubMed Central

    Wyroba, E.; Kwaśniak, P.; Miller, K.; Kobyłecki, K.; Osińska, M.

    2016-01-01

    Protein products of paralogous genes resulting from whole genome duplication may acquire new functions. The role of post-translational modifications (PTM) in proper targeting of Paramecium Rab7b paralogue (distinct from that of Rab7a directly involved in phagocytosis) was studied using point mutagenesis, proteomic analysis and double immunofluorescence after in vivo electroporation of the mutagenized protein. Here we show that substitution of Thr200 by Ala diminished the incorporation of [P32] by 37% and of [C14-]UDP-glucose by 24% into recombinant Rab7b_200 in comparison to the non-mutagenized control. Double confocal imaging revealed that Rab7b_200 was mistargeted upon electroporation into living cells in contrast to non-mutagenized recombinant Rab7b correctly incorporated in the cytostome area. Using nano LC-MS/MS to compare the peptide map of Rab7b with that after deglycosylation with a mixture of five enzymes of different specificity we identified a peptide ion at m/z=677.63+ representing a glycan group attached to Thr200. Based on its mass and quantitative assays with [P32] and [C14]UDP-glucose, the suggested composition of the adduct attached to Thr200 is (Hex)1(HexNAc)1(Phos)3 or (HexNAc)1 (Deoxyhexose)1 (Phos)1 (HexA)1. These data indicate that PTM of Thr200 located in the hypervariable C-region of Paramecium octaurelia Rab7b is crucial for the proper localization/function of this protein. Moreover, the two Rab7 paralogues differ also in another PTM: substantially more phosphorylated amino acid residues are in Rab7b than in Rab7a. PMID:27349314

  3. Site-directed mutagenesis, in vivo electroporation and mass spectrometry in search for determinants of the subcellular targeting of Rab7b paralogue in the model eukaryote Paramecium octaurelia.

    PubMed

    Wyroba, E; Kwaśniak, P; Miller, K; Kobyłecki, K; Osińska, M

    2016-01-01

    Protein products of the paralogous genes resulting from the whole genome duplication may acquire new function. The role of post-translational modifications (PTM) in proper targeting of Paramecium Rab7b paralogue - distinct from that of Rab7a directly involved in phagocytosis - was studied using point mutagenesis, proteomic analysis and double immunofluorescence after in vivo electroporation of the mutagenized protein. Here we show that substitution of Thr200 by Ala200 resulted in diminished incorporation of [P32] by 37.4% and of 32 [C14-]UDP-glucose by 24%, respectively, into recombinant Rab7b_200 in comparison to the non-mutagenized control. Double confocal imaging revealed that Rab7b_200 was mistargeted upon electroporation into living cells contrary to non- mutagenized recombinant Rab7b correctly incorporated in the cytostome area. We identified the peptide ion at m/z=677.63+ characteristic for the glycan group attached to Thr200 in Rab7b using nano LC-MS/MS and comparing the peptide map of this protein with that after deglycosylation with the mixture of five enzymes of different specificity. Based on the mass of this peptide ion and quantitative radioactive assays with [P32]and  [C14-]UDP- glucose, the suggested composition of the adduct attached to Thr200 might be (Hex)1(HexNAc)1(Phos)3 or (HexNAc)1 (Deoxyhexose)1 (Phos)1 (HexA)1. These data indicate that PTM of Thr200 located in the hypervariable C-region of Rab7b in Paramecium is crucial for the proper localization/function of this protein. Moreover, these proteins differ also in other PTM: the number of phosphorylated amino acids in Rab7b is much higher than in Rab7a. PMID:27349314

  4. Site-directed mutagenesis, in vivo electroporation and mass spectrometry in search for determinants of the subcellular targeting of Rab7b paralogue in the model eukaryote Paramecium octaurelia.

    PubMed

    Wyroba, E; Kwaśniak, P; Miller, K; Kobyłecki, K; Osińska, M

    2016-01-01

    Protein products of the paralogous genes resulting from the whole genome duplication may acquire new function. The role of post-translational modifications (PTM) in proper targeting of Paramecium Rab7b paralogue - distinct from that of Rab7a directly involved in phagocytosis - was studied using point mutagenesis, proteomic analysis and double immunofluorescence after in vivo electroporation of the mutagenized protein. Here we show that substitution of Thr200 by Ala200 resulted in diminished incorporation of [P32] by 37.4% and of 32 [C14-]UDP-glucose by 24%, respectively, into recombinant Rab7b_200 in comparison to the non-mutagenized control. Double confocal imaging revealed that Rab7b_200 was mistargeted upon electroporation into living cells contrary to non- mutagenized recombinant Rab7b correctly incorporated in the cytostome area. We identified the peptide ion at m/z=677.63+ characteristic for the glycan group attached to Thr200 in Rab7b using nano LC-MS/MS and comparing the peptide map of this protein with that after deglycosylation with the mixture of five enzymes of different specificity. Based on the mass of this peptide ion and quantitative radioactive assays with [P32]and  [C14-]UDP- glucose, the suggested composition of the adduct attached to Thr200 might be (Hex)1(HexNAc)1(Phos)3 or (HexNAc)1 (Deoxyhexose)1 (Phos)1 (HexA)1. These data indicate that PTM of Thr200 located in the hypervariable C-region of Rab7b in Paramecium is crucial for the proper localization/function of this protein. Moreover, these proteins differ also in other PTM: the number of phosphorylated amino acids in Rab7b is much higher than in Rab7a.

  5. High DRC Levels Are Associated with Let-7b Overexpression in Women with Breast Cancer

    PubMed Central

    Encarnación, Jarline; Ortiz, Carmen; Vergne, Ralphdy; Vargas, Wanda; Coppola, Domenico; Matta, Jaime L.

    2016-01-01

    Nucleotide Excision Repair (NER) is a critical pathway involved in breast cancer (BC). We have previously published that a low DNA repair capacity (DRC) is associated with a higher risk of BC in Puerto Rican women. Let-7b belongs to a miRNA family with tumor suppressor activity that targets oncogenes. We isolated miRNAs from plasma of 153 Puerto Rican women with and without BC. DRC was measured in lymphocytes by means of a host cell reactivation assay. These women were divided into four groups according to their DRC level: High (>3.8%) and low (<3.8%). The four groups consisted of BC patients with high (n = 35) and low (n = 43) DRC and controls with high (n = 39) and low (n = 36) DRC. Epidemiologic data were collected at initial BC diagnosis and almost five years after diagnosis. A significant difference in Let-7b expression was found in BC patients with high DRC versus the remaining groups (p < 0.001). Thus, our data reveal a possible role of Let-7b on DRC during breast carcinogenesis. Our study is innovative because it provides the first evidence that Let-7b may play role in DRC regulation (through the NER repair pathway) in BC. PMID:27271599

  6. Let-7b promotes alpaca hair growth via transcriptional repression of TGFβR I.

    PubMed

    Yan, Shen; Yu, Zhang; Ning, Liu; Hai-Dong, Wang; Jian-Shan, Xie; Shu-Yuan, Gao; Jia-Qi, Cheng; Xiu-Ju, Yu; Ting, Wang; Chang-Sheng, Dong; Xiao-Yan, He

    2016-02-10

    The young male alpaca ear and the back skins were used to investigate the effect of transforming growth factor receptor-β I (TGFβR I) on alpaca hair follicles and hair growth. The expression level and location of TGFβR I in alpaca ear and dorsal skin were detected through real-time quantitative PCR (RT-PCR) and paraffin section immunohistochemical technique (ICC-P). The results shown TGFβR I was lower expression in back skin compared to ear skin and the mean density of the positive reaction in ear skin was significantly higher than back skin. The targeted relationship with let-7b was detected using the dual-luciferase reporter vector of TGFβR I, which showed a significant target relationship between let-7b and TGFβR I. After transfection with let-7b eukaryotic expression vector, the relative mRNA expression of TGFβR I in alpaca skin fibroblasts did not differ, while the relative protein level was significantly decreased. In summary, a higher TGFβR I expression level in the ear skin suggests that TGFβR I may inhibit coat hair elongation. Further studies showed TGFβR I protein was downregulated by let-7b through transcriptional repression.

  7. 29 CFR 794.140 - Compensation requirements for a workweek under section 7(b)(3).

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 3 2012-07-01 2012-07-01 false Compensation requirements for a workweek under section 7(b)(3). 794.140 Section 794.140 Labor Regulations Relating to Labor (Continued) WAGE AND HOUR DIVISION, DEPARTMENT OF LABOR STATEMENTS OF GENERAL POLICY OR INTERPRETATION NOT DIRECTLY RELATED TO REGULATIONS PARTIAL OVERTIME EXEMPTION FOR EMPLOYEES...

  8. Whole-Genome Sequence of Stenotrophomonas maltophilia ZBG7B Reveals Its Biotechnological Potential

    PubMed Central

    Chong, Teik-Min; Adrian, Tan-Guan-Sheng; Kher, Heng Leong; Hong, Kar-Wai; Grandclément, Catherine; Faure, Denis; Yin, Wai-Fong; Dessaux, Yves

    2015-01-01

    Stenotrophomonas maltophilia ZBG7B was isolated from vineyard soil of Zellenberg, France. Here, we present the draft genome sequence of this bacterial strain, which has facilitated the prediction of function for several genes encoding biotechnologically important enzymes, such as xylosidase, xylanase, laccase, and chitinase. PMID:26659682

  9. 46 CFR 151.50-86 - Alkyl (C7-C9) nitrates.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 5 2012-10-01 2012-10-01 false Alkyl (C7-C9) nitrates. 151.50-86 Section 151.50-86 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES BARGES CARRYING BULK LIQUID HAZARDOUS MATERIAL CARGOES Special Requirements § 151.50-86 Alkyl (C7-C9) nitrates....

  10. 46 CFR 153.560 - Special requirements for Alkyl (C7-C9) nitrates.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 5 2012-10-01 2012-10-01 false Special requirements for Alkyl (C7-C9) nitrates. 153.560 Section 153.560 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS... Equipment Special Requirements § 153.560 Special requirements for Alkyl (C7-C9) nitrates. (a) The...

  11. 46 CFR 151.50-86 - Alkyl (C7-C9) nitrates.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 5 2014-10-01 2014-10-01 false Alkyl (C7-C9) nitrates. 151.50-86 Section 151.50-86 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES BARGES CARRYING BULK LIQUID HAZARDOUS MATERIAL CARGOES Special Requirements § 151.50-86 Alkyl (C7-C9) nitrates....

  12. 46 CFR 151.50-86 - Alkyl (C7-C9) nitrates.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 5 2011-10-01 2011-10-01 false Alkyl (C7-C9) nitrates. 151.50-86 Section 151.50-86 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES BARGES CARRYING BULK LIQUID HAZARDOUS MATERIAL CARGOES Special Requirements § 151.50-86 Alkyl (C7-C9) nitrates....

  13. 46 CFR 153.560 - Special requirements for Alkyl (C7-C9) nitrates.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 5 2011-10-01 2011-10-01 false Special requirements for Alkyl (C7-C9) nitrates. 153.560 Section 153.560 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS... Equipment Special Requirements § 153.560 Special requirements for Alkyl (C7-C9) nitrates. (a) The...

  14. 46 CFR 151.50-86 - Alkyl (C7-C9) nitrates.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 5 2010-10-01 2010-10-01 false Alkyl (C7-C9) nitrates. 151.50-86 Section 151.50-86 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES BARGES CARRYING BULK LIQUID HAZARDOUS MATERIAL CARGOES Special Requirements § 151.50-86 Alkyl (C7-C9) nitrates....

  15. 46 CFR 153.560 - Special requirements for Alkyl (C7-C9) nitrates.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 5 2014-10-01 2014-10-01 false Special requirements for Alkyl (C7-C9) nitrates. 153.560 Section 153.560 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS... Equipment Special Requirements § 153.560 Special requirements for Alkyl (C7-C9) nitrates. (a) The...

  16. 46 CFR 153.560 - Special requirements for Alkyl (C7-C9) nitrates.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 5 2010-10-01 2010-10-01 false Special requirements for Alkyl (C7-C9) nitrates. 153.560 Section 153.560 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS... Equipment Special Requirements § 153.560 Special requirements for Alkyl (C7-C9) nitrates. (a) The...

  17. 46 CFR 153.560 - Special requirements for Alkyl (C7-C9) nitrates.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 5 2013-10-01 2013-10-01 false Special requirements for Alkyl (C7-C9) nitrates. 153.560 Section 153.560 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS... Equipment Special Requirements § 153.560 Special requirements for Alkyl (C7-C9) nitrates. (a) The...

  18. 46 CFR 151.50-86 - Alkyl (C7-C9) nitrates.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 5 2013-10-01 2013-10-01 false Alkyl (C7-C9) nitrates. 151.50-86 Section 151.50-86 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES BARGES CARRYING BULK LIQUID HAZARDOUS MATERIAL CARGOES Special Requirements § 151.50-86 Alkyl (C7-C9) nitrates....

  19. Distal hereditary motor neuropathy type 7B with Dynactin 1 mutation.

    PubMed

    Hwang, Sun Hee; Kim, Eun Ja; Hong, Young Bin; Joo, Jaesoon; Kim, Sung Min; Nam, Soo Hyun; Hong, Hyun Dae; Kim, Seung Hyun; Oh, Kiwook; Lim, Jeong-Geun; Cho, Jeong Hee; Chung, Ki Wha; Choi, Byung-Ok

    2016-10-01

    Mutations in the Dynactin 1 (DCTN1) gene have been demonstrated to result in various neurodegenerative diseases, including distal hereditary motor neuropathy type 7B (dHMN7B), Perry syndrome, amyotrophic lateral sclerosis and amyotrophic lateral sclerosis‑frontotemporal dementia. However, since the first dHMN7B patient with a DCTN1 mutation was described in 2003, to the best of our knowledge no further cases have been reported. In the present study, the DCTN1 p.G59S mutation was identified in two unrelated families from a total of 24 Korean families with dHMN, by whole exome sequencing. Codon 59 appears to be the mutational hot spot in the DCTN1 gene, as all described dHMN7B patients to date have harbored an identical p.G59S mutation. The families of the present study with the DCTN1 mutation had a milder disease with a later onset compared with the previously described patients. No affected family members exhibited facial muscle weakness or bulbar involvement. One family member demonstrated vocal cord palsy as the initial sign of disease; however, in the other family hand muscle weakness was the first major symptom. No affected patients demonstrated sensory loss or upper motor neuron involvements. Although this is only the second report of dHMN7B resulting from a DCTN1 mutation, the frequency of the DCTN1 mutation was not low in the Korean population examined, and clinical heterogeneities were observed in patients with the DCTN1 mutation. Therefore, it may be beneficial to screen all dHMN patients for the DCTN1 mutation.

  20. Distal hereditary motor neuropathy type 7B with Dynactin 1 mutation.

    PubMed

    Hwang, Sun Hee; Kim, Eun Ja; Hong, Young Bin; Joo, Jaesoon; Kim, Sung Min; Nam, Soo Hyun; Hong, Hyun Dae; Kim, Seung Hyun; Oh, Kiwook; Lim, Jeong-Geun; Cho, Jeong Hee; Chung, Ki Wha; Choi, Byung-Ok

    2016-10-01

    Mutations in the Dynactin 1 (DCTN1) gene have been demonstrated to result in various neurodegenerative diseases, including distal hereditary motor neuropathy type 7B (dHMN7B), Perry syndrome, amyotrophic lateral sclerosis and amyotrophic lateral sclerosis‑frontotemporal dementia. However, since the first dHMN7B patient with a DCTN1 mutation was described in 2003, to the best of our knowledge no further cases have been reported. In the present study, the DCTN1 p.G59S mutation was identified in two unrelated families from a total of 24 Korean families with dHMN, by whole exome sequencing. Codon 59 appears to be the mutational hot spot in the DCTN1 gene, as all described dHMN7B patients to date have harbored an identical p.G59S mutation. The families of the present study with the DCTN1 mutation had a milder disease with a later onset compared with the previously described patients. No affected family members exhibited facial muscle weakness or bulbar involvement. One family member demonstrated vocal cord palsy as the initial sign of disease; however, in the other family hand muscle weakness was the first major symptom. No affected patients demonstrated sensory loss or upper motor neuron involvements. Although this is only the second report of dHMN7B resulting from a DCTN1 mutation, the frequency of the DCTN1 mutation was not low in the Korean population examined, and clinical heterogeneities were observed in patients with the DCTN1 mutation. Therefore, it may be beneficial to screen all dHMN patients for the DCTN1 mutation. PMID:27573046

  1. Mutational analysis of ATP7B in Chinese Wilson disease patients.

    PubMed

    Hua, Rui; Hua, Fang; Jiao, Yonggeng; Pan, Yu; Yang, Xu; Peng, Shanshan; Niu, Junqi

    2016-01-01

    Wilson Disease (WD) is an inborn error of copper metabolism inherited in an autosomal recessive manner caused by the mutations in the P-type ATPase gene (ATP7B). In this study, we screen and detect the mutations of the ATP7B gene in unrelated Chinese WD patients. A total of 68 individuals from ten provinces of China with WD were recruited. Of them, 43 were males and 25 were females, and their onset ages were from 1 to 48 years with a median onset age of 22.2 years. All the exons and exon/intron boundaries of ATP7B gene of the patients were sequenced and aligned to the referred ATP7B gene sequence. The results suggested that 66 of the 68 patents carried with at least one mutation and 48 different mutations were identified including 34 missense, one synonymous, two nonsense, two splicing, and nine frameshift mutations (five insertion and four deletion). Among these mutations, c.2333G>T, c.2310C>G, c.2975C>T, and c.3443T>C were the most prevalent mutants and c.2310C>G always linked with c.2333G>T. The eighth, 11(th), and 18(th) exons carried more mutations (6/48, 5/48, and 5/48, respectively) than others. After comparing with the mutations reported previously, 22 out of the 48 mutations were identified as novel mutations. A popular algorithm, Polyphen-2, was used to predict the effects of the amino-acid substitution due to the mutations on the structure and function of ATP7B function and the predicted results indicated that all the missense mutations were unfavorable except c.121A>G and c.748G>A. Phenotype/genotype correlation analysis suggested that the patients with c.2975C>T or c.3809A>G often presented WD features before 12 years old while the patients with c.3443T>C almost presented WD after 12 years old. This is the first time to identify the common mutations contributing to early onset age in Chinese WD patients. Our study will broaden our knowledge about ATP7B mutations in WD patients.

  2. Mutational analysis of ATP7B in Chinese Wilson disease patients.

    PubMed

    Hua, Rui; Hua, Fang; Jiao, Yonggeng; Pan, Yu; Yang, Xu; Peng, Shanshan; Niu, Junqi

    2016-01-01

    Wilson Disease (WD) is an inborn error of copper metabolism inherited in an autosomal recessive manner caused by the mutations in the P-type ATPase gene (ATP7B). In this study, we screen and detect the mutations of the ATP7B gene in unrelated Chinese WD patients. A total of 68 individuals from ten provinces of China with WD were recruited. Of them, 43 were males and 25 were females, and their onset ages were from 1 to 48 years with a median onset age of 22.2 years. All the exons and exon/intron boundaries of ATP7B gene of the patients were sequenced and aligned to the referred ATP7B gene sequence. The results suggested that 66 of the 68 patents carried with at least one mutation and 48 different mutations were identified including 34 missense, one synonymous, two nonsense, two splicing, and nine frameshift mutations (five insertion and four deletion). Among these mutations, c.2333G>T, c.2310C>G, c.2975C>T, and c.3443T>C were the most prevalent mutants and c.2310C>G always linked with c.2333G>T. The eighth, 11(th), and 18(th) exons carried more mutations (6/48, 5/48, and 5/48, respectively) than others. After comparing with the mutations reported previously, 22 out of the 48 mutations were identified as novel mutations. A popular algorithm, Polyphen-2, was used to predict the effects of the amino-acid substitution due to the mutations on the structure and function of ATP7B function and the predicted results indicated that all the missense mutations were unfavorable except c.121A>G and c.748G>A. Phenotype/genotype correlation analysis suggested that the patients with c.2975C>T or c.3809A>G often presented WD features before 12 years old while the patients with c.3443T>C almost presented WD after 12 years old. This is the first time to identify the common mutations contributing to early onset age in Chinese WD patients. Our study will broaden our knowledge about ATP7B mutations in WD patients. PMID:27398169

  3. Mutational analysis of ATP7B in Chinese Wilson disease patients

    PubMed Central

    Hua, Rui; Hua, Fang; Jiao, Yonggeng; Pan, Yu; Yang, Xu; Peng, Shanshan; Niu, Junqi

    2016-01-01

    Wilson Disease (WD) is an inborn error of copper metabolism inherited in an autosomal recessive manner caused by the mutations in the P-type ATPase gene (ATP7B). In this study, we screen and detect the mutations of the ATP7B gene in unrelated Chinese WD patients. A total of 68 individuals from ten provinces of China with WD were recruited. Of them, 43 were males and 25 were females, and their onset ages were from 1 to 48 years with a median onset age of 22.2 years. All the exons and exon/intron boundaries of ATP7B gene of the patients were sequenced and aligned to the referred ATP7B gene sequence. The results suggested that 66 of the 68 patents carried with at least one mutation and 48 different mutations were identified including 34 missense, one synonymous, two nonsense, two splicing, and nine frameshift mutations (five insertion and four deletion). Among these mutations, c.2333G>T, c.2310C>G, c.2975C>T, and c.3443T>C were the most prevalent mutants and c.2310C>G always linked with c.2333G>T. The eighth, 11th, and 18th exons carried more mutations (6/48, 5/48, and 5/48, respectively) than others. After comparing with the mutations reported previously, 22 out of the 48 mutations were identified as novel mutations. A popular algorithm, Polyphen-2, was used to predict the effects of the amino-acid substitution due to the mutations on the structure and function of ATP7B function and the predicted results indicated that all the missense mutations were unfavorable except c.121A>G and c.748G>A. Phenotype/genotype correlation analysis suggested that the patients with c.2975C>T or c.3809A>G often presented WD features before 12 years old while the patients with c.3443T>C almost presented WD after 12 years old. This is the first time to identify the common mutations contributing to early onset age in Chinese WD patients. Our study will broaden our knowledge about ATP7B mutations in WD patients. PMID:27398169

  4. MicroRNAs let-7b/i suppress human glioma cell invasion and migration by targeting IKBKE directly

    SciTech Connect

    Tian, Yuan; Hao, Shaobo; Ye, Minhua; Zhang, Anling; Nan, Yang; Wang, Guangxiu; Jia, Zhifan; Yu, Kai; Guo, Lianmei; Pu, Peiyu; Huang, Qiang; Zhong, Yue

    2015-03-06

    We demonstrated that IKBKE is overexpressed in human gliomas and that the downregulation of IKBKE markedly inhibits the proliferative and invasive abilities of glioma cells, which is consistent with the results reported by several different research groups. Therefore, IKBKE represents a promising therapeutic target for the treatment of glioma. In the present study, we verified that the microRNAs let-7b and let-7i target IKBKE through luciferase assays and found that let-7b/i mimics can knock down IKBKE and upregulate E-cadherin through western blot analysis. Moreover, the expression levels of let-7b/i were significantly lower in glioma cell lines than that in normal brain tissues, as determined by quantitative real-time PCR. Furthermore, let-7b/i inhibit the invasion and migration of glioma cells, as determined through wound healing and Transwell assays. The above-mentioned data suggest that let-7b/i inhibit the invasive ability of glioma cells by directly downregulating IKBKE and indirectly upregulating E-cadherin. - Highlights: • Let-7b and let-7i are downregulated in glioma cell lines. • IKBKE is a target gene of let-7b/i. • Let-7b/i inhibit the invasion and migration of glioma cells. • Let-7b/i upregulate E-cadherin by downregulating IKBKE.

  5. Kepler's optical phase curve of the exoplanet HAT-P-7b.

    PubMed

    Borucki, W J; Koch, D; Jenkins, J; Sasselov, D; Gilliland, R; Batalha, N; Latham, D W; Caldwell, D; Basri, G; Brown, T; Christensen-Dalsgaard, J; Cochran, W D; DeVore, E; Dunham, E; Dupree, A K; Gautier, T; Geary, J; Gould, A; Howell, S; Kjeldsen, H; Lissauer, J; Marcy, G; Meibom, S; Morrison, D; Tarter, J

    2009-08-01

    Ten days of photometric data were obtained during the commissioning phase of the Kepler mission, including data for the previously known giant transiting exoplanet HAT-P-7b. The data for HAT-P-7b show a smooth rise and fall of light from the planet as it orbits its star, punctuated by a drop of 130 +/- 11 parts per million in flux when the planet passes behind its star. We interpret this as the phase variation of the dayside thermal emission plus reflected light from the planet as it orbits its star and is occulted. The depth of the occultation is similar in photometric precision to the detection of a transiting Earth-size planet for which the mission was designed.

  6. Kepler's optical phase curve of the exoplanet HAT-P-7b.

    PubMed

    Borucki, W J; Koch, D; Jenkins, J; Sasselov, D; Gilliland, R; Batalha, N; Latham, D W; Caldwell, D; Basri, G; Brown, T; Christensen-Dalsgaard, J; Cochran, W D; DeVore, E; Dunham, E; Dupree, A K; Gautier, T; Geary, J; Gould, A; Howell, S; Kjeldsen, H; Lissauer, J; Marcy, G; Meibom, S; Morrison, D; Tarter, J

    2009-08-01

    Ten days of photometric data were obtained during the commissioning phase of the Kepler mission, including data for the previously known giant transiting exoplanet HAT-P-7b. The data for HAT-P-7b show a smooth rise and fall of light from the planet as it orbits its star, punctuated by a drop of 130 +/- 11 parts per million in flux when the planet passes behind its star. We interpret this as the phase variation of the dayside thermal emission plus reflected light from the planet as it orbits its star and is occulted. The depth of the occultation is similar in photometric precision to the detection of a transiting Earth-size planet for which the mission was designed. PMID:19661420

  7. A kinetic study of Trichoderma reesei Cel7B catalyzed cellulose hydrolysis.

    PubMed

    Song, Xiangfei; Zhang, Shujun; Wang, Yefei; Li, Jingwen; He, Chunyan; Yao, Lishan

    2016-06-01

    One prominent feature of Trichoderma reesei (Tr) endoglucanases catalyzed cellulose hydrolysis is that the reaction slows down quickly after it starts (within minutes). But the mechanism of the slowdown is not well understood. A structural model of Tr- Cel7B catalytic domain bound to cellulose was built computationally and the potentially important binding residues were identified and tested experimentally. The 13 tested mutants show different binding properties in the adsorption to phosphoric acid swollen cellulose and filter paper. Though the partitioning parameter to filter paper is about 10 times smaller than that to phosphoric acid swollen cellulose, a positive correlation is shown for two substrates. The kinetic studies show that the reactions slow down quickly for both substrates. This slowdown is not correlated to the binding constant but anticorrelated to the enzyme initial activity. The amount of reducing sugars released after 24h by Cel7B in phosphoric acid swollen cellulose, Avicel and filter paper cellulose hydrolysis is correlated with the enzyme activity against a soluble substrate p-nitrophenyl lactoside. Six of the 13 tested mutants, including N47A, N52D, S99A, N323D, S324A, and S346A, yield ∼15-35% more reducing sugars than the wild type (WT) Cel7B in phosphoric acid swollen cellulose and filter paper hydrolysis. This study reveals that the slowdown of the reaction is not due to the binding of the enzyme to cellulose. The activity of Tr- Cel7B against the insoluble substrate cellulose is determined by the enzyme's capability in hydrolyzing the soluble substrate. PMID:27178789

  8. A kinetic study of Trichoderma reesei Cel7B catalyzed cellulose hydrolysis.

    PubMed

    Song, Xiangfei; Zhang, Shujun; Wang, Yefei; Li, Jingwen; He, Chunyan; Yao, Lishan

    2016-06-01

    One prominent feature of Trichoderma reesei (Tr) endoglucanases catalyzed cellulose hydrolysis is that the reaction slows down quickly after it starts (within minutes). But the mechanism of the slowdown is not well understood. A structural model of Tr- Cel7B catalytic domain bound to cellulose was built computationally and the potentially important binding residues were identified and tested experimentally. The 13 tested mutants show different binding properties in the adsorption to phosphoric acid swollen cellulose and filter paper. Though the partitioning parameter to filter paper is about 10 times smaller than that to phosphoric acid swollen cellulose, a positive correlation is shown for two substrates. The kinetic studies show that the reactions slow down quickly for both substrates. This slowdown is not correlated to the binding constant but anticorrelated to the enzyme initial activity. The amount of reducing sugars released after 24h by Cel7B in phosphoric acid swollen cellulose, Avicel and filter paper cellulose hydrolysis is correlated with the enzyme activity against a soluble substrate p-nitrophenyl lactoside. Six of the 13 tested mutants, including N47A, N52D, S99A, N323D, S324A, and S346A, yield ∼15-35% more reducing sugars than the wild type (WT) Cel7B in phosphoric acid swollen cellulose and filter paper hydrolysis. This study reveals that the slowdown of the reaction is not due to the binding of the enzyme to cellulose. The activity of Tr- Cel7B against the insoluble substrate cellulose is determined by the enzyme's capability in hydrolyzing the soluble substrate.

  9. Improving the activity of Trichoderma reesei cel7B through stabilizing the transition state.

    PubMed

    Wang, Yefei; Song, Xiangfei; Zhang, Shujun; Li, Jingwen; Shu, Zhiyu; He, Chunyan; Huang, Qingshan; Yao, Lishan

    2016-06-01

    Trichoderma reesei (Tr.) cellulases, which convert cellulose to reducing sugars, are a promising catalyst used in the lignocellulosic biofuel production. Improving Tr. cellulases activity, though very difficult, is highly desired due to the recalcitrance of lignocellulose. Meanwhile, it is preferable to enhance the cellulase's promiscuity so that substrates other than cellulose can also be hydrolyzed. In this work, an attempt is made to improve the catalytic activity of a major endogluanase Tr. Cel7B against xylan which crosslinks with cellulose in lignocellulose. By using quantum mechanics/molecular mechanics (QM/MM) molecular dynamics (MD) simulations, the transition state of the xylo-oligosaccharide hydrolysis is identified. Then, mutations are introduced and their effect on the transition state stabilization is ranked based on the free energy calculations. Seven top ranked mutants are evaluated experimentally. Three mutants A208Q, A222D, and G230R show a higher activity than the wild-type Tr. Cel7B in the hydrolysis of xylan (by up to 47%) as well as filter paper (by up to 50%). The combination of the single mutants can further improve the enzyme activity. Our work demonstrates that the free energy method is effective in engineering the Tr. Cel7B activity against xylan and cellulose, and thus may also be useful for improving the activity of other Tr. cellulases. Biotechnol. Bioeng. 2016;113: 1171-1177. © 2015 Wiley Periodicals, Inc. PMID:26616246

  10. AP180-mediated trafficking of Vamp7B limits homotypic fusion of Dictyostelium contractile vacuoles.

    PubMed

    Wen, Yujia; Stavrou, Irene; Bersuker, Kirill; Brady, Rebecca J; De Lozanne, Arturo; O'Halloran, Theresa J

    2009-10-01

    Clathrin-coated vesicles play an established role in endocytosis from the plasma membrane, but they are also found on internal organelles. We examined the composition of clathrin-coated vesicles on an internal organelle responsible for osmoregulation, the Dictyostelium discoideum contractile vacuole. Clathrin puncta on contractile vacuoles contained multiple accessory proteins typical of plasma membrane-coated pits, including AP2, AP180, and epsin, but not Hip1r. To examine how these clathrin accessory proteins influenced the contractile vacuole, we generated cell lines that carried single and double gene knockouts in the same genetic background. Single or double mutants that lacked AP180 or AP2 exhibited abnormally large contractile vacuoles. The enlarged contractile vacuoles in AP180-null mutants formed because of excessive homotypic fusion among contractile vacuoles. The SNARE protein Vamp7B was mislocalized and enriched on the contractile vacuoles of AP180-null mutants. In vitro assays revealed that AP180 interacted with the cytoplasmic domain of Vamp7B. We propose that AP180 directs Vamp7B into clathrin-coated vesicles on contractile vacuoles, creating an efficient mechanism for regulating the internal distribution of fusion-competent SNARE proteins and limiting homotypic fusions among contractile vacuoles. Dictyostelium contractile vacuoles offer a valuable system to study clathrin-coated vesicles on internal organelles within eukaryotic cells.

  11. Non-ceruloplasmin bound copper and ATP7B gene variants in Alzheimer's disease.

    PubMed

    Squitti, R; Siotto, M; Arciello, M; Rossi, L

    2016-09-01

    ATP7B, a protein mainly expressed in the hepatocytes, is a copper chaperone that loads the metal into the serum copper-protein ceruloplasmin during its synthesis and also escorts superfluous copper into the bile, by a sophisticated trafficking mechanism. Impaired function of this ATPase is associated with a well-known inborn error of copper metabolism, Wilson's disease (WD). Several mutations of ATP7B are known, involving different regions of the protein, thus resulting in a plethora of phenotypes in WD patients. It is a consolidated notion that copper dysmetabolism occurs in Alzheimer's disease (AD) as well. Besides the molecular mechanisms relating copper to the protein hallmarks of this disease and neurodegeneration, more recently the observation that a free-copper in the serum, not bound to ceruloplasmin (non-Cp-Cu), characterizes AD patients, prompted our research to identify possible genetic defects of the ATP7B gene in AD patients. Four specific single nucleotide polymorphisms and a WD rare mutation have a statistical association with AD. They contribute to characterize a copper subtype of AD. Additional facets of this AD phenotype, typified by higher levels of non-Cp-Cu, are presented and discussed in the framework of copper failure as an accelerator risk factor of neurological disorders with different aetiology. PMID:27499330

  12. Non-ceruloplasmin bound copper and ATP7B gene variants in Alzheimer's disease.

    PubMed

    Squitti, R; Siotto, M; Arciello, M; Rossi, L

    2016-09-01

    ATP7B, a protein mainly expressed in the hepatocytes, is a copper chaperone that loads the metal into the serum copper-protein ceruloplasmin during its synthesis and also escorts superfluous copper into the bile, by a sophisticated trafficking mechanism. Impaired function of this ATPase is associated with a well-known inborn error of copper metabolism, Wilson's disease (WD). Several mutations of ATP7B are known, involving different regions of the protein, thus resulting in a plethora of phenotypes in WD patients. It is a consolidated notion that copper dysmetabolism occurs in Alzheimer's disease (AD) as well. Besides the molecular mechanisms relating copper to the protein hallmarks of this disease and neurodegeneration, more recently the observation that a free-copper in the serum, not bound to ceruloplasmin (non-Cp-Cu), characterizes AD patients, prompted our research to identify possible genetic defects of the ATP7B gene in AD patients. Four specific single nucleotide polymorphisms and a WD rare mutation have a statistical association with AD. They contribute to characterize a copper subtype of AD. Additional facets of this AD phenotype, typified by higher levels of non-Cp-Cu, are presented and discussed in the framework of copper failure as an accelerator risk factor of neurological disorders with different aetiology.

  13. Improving the activity of Trichoderma reesei cel7B through stabilizing the transition state.

    PubMed

    Wang, Yefei; Song, Xiangfei; Zhang, Shujun; Li, Jingwen; Shu, Zhiyu; He, Chunyan; Huang, Qingshan; Yao, Lishan

    2016-06-01

    Trichoderma reesei (Tr.) cellulases, which convert cellulose to reducing sugars, are a promising catalyst used in the lignocellulosic biofuel production. Improving Tr. cellulases activity, though very difficult, is highly desired due to the recalcitrance of lignocellulose. Meanwhile, it is preferable to enhance the cellulase's promiscuity so that substrates other than cellulose can also be hydrolyzed. In this work, an attempt is made to improve the catalytic activity of a major endogluanase Tr. Cel7B against xylan which crosslinks with cellulose in lignocellulose. By using quantum mechanics/molecular mechanics (QM/MM) molecular dynamics (MD) simulations, the transition state of the xylo-oligosaccharide hydrolysis is identified. Then, mutations are introduced and their effect on the transition state stabilization is ranked based on the free energy calculations. Seven top ranked mutants are evaluated experimentally. Three mutants A208Q, A222D, and G230R show a higher activity than the wild-type Tr. Cel7B in the hydrolysis of xylan (by up to 47%) as well as filter paper (by up to 50%). The combination of the single mutants can further improve the enzyme activity. Our work demonstrates that the free energy method is effective in engineering the Tr. Cel7B activity against xylan and cellulose, and thus may also be useful for improving the activity of other Tr. cellulases. Biotechnol. Bioeng. 2016;113: 1171-1177. © 2015 Wiley Periodicals, Inc.

  14. CoRoT-7b: Convection in a Tidally Locked Planet

    NASA Astrophysics Data System (ADS)

    Noack, Lena; Stamenkovic, Vlada; Wagner, Frank W.; Sohl, Frank; Breuer, Doris

    2010-05-01

    The number of terrestrial extrasolar planets found in the past few years is increasing rapidly. Some have masses ranging from 2 to 10 Earth masses, and the habitability of these planets is widely discussed in the planetary community. Due to observational limitations we will mostly be able to observe planets that are very close to its host star, resulting in a potentially tidally locked orbit. Our goal is to investigate if such planets can be habitable at all. But to do so, we first have to understand the convection behaviour of such planets. In this work we model the mantle convection of the recently discovered exoplanet CoRoT-7b [1], which is a planet believed to be tidally locked. The extreme intense insolation in the vicinity of its host star heats the day-side of CoRoT-7b, leading to surface temperatures about 2000 Kelvin higher than on the night-side [1]. CoRoT-7b is about 5 times more massive than the Earth and predominantly composed of dry silicate rock similar to Earth's Moon. A central iron core, if present, would be relatively small [2] with a core mass fraction of no more than 15 wt%. The mantle convection is modelled in a spherical shell [3] using a temperature- and pressure-dependent viscosity. We use a radioactive heat source density similar to present Earth. Coriolis forces are neglected and we assume that CoRoT-7b has no atmosphere. The results show that the lower mantle above the core-mantle boundary is in a more sluggish convection regime as a consequence of the viscosity increase with pressure. Depending on the strength of the viscosity increase, even a so-called low-lid [4] can form and conductive heat transport dominates from the core to the upper part of the mantle. The thermal state of such a deeply situated, conductive lower mantle of CoRoT-7b is not much influenced by the strongly laterally varying surface temperature. However, the temperatures of the upper convecting mantle are found to strongly vary from one side of the planet to the

  15. Hemolytic anemia as first presentation of Wilson's disease with uncommon ATP7B mutation.

    PubMed

    Ye, Xing-Nong; Mao, Li-Ping; Lou, Yin-Jun; Tong, Hong-Yan

    2015-01-01

    Wilson's disease (WD) is a rare inherited disorder of copper metabolism and the main manifestations are liver and brain disorders. Hemolytic anemia is an unusual complication of WD. We describe a 15-year-old girl who developed hemolytic anemia as the first manifestation of Wilson's disease. An Arg952Lys mutation was found in exon 12 of the ATP7B gene, which is uncommon among Chinese Han individuals. From this case and reviews, we can achieve a better understanding of WD. Besides, we may conclude that the probable diagnosis of WD should be considered in young patients with unexplained hemolytic anemia, especially in patients with hepatic and/or neurologic disorder.

  16. "Invar"-like behavior in compressed Fe7C3 with implication for deep carbon cycle

    NASA Astrophysics Data System (ADS)

    Liu, J.; Li, J.; Ikuta, D.

    2014-12-01

    Iron carbide Fe7C3 has recently emerged as a leading candidate component of the inner core because it is likely the first phase to solidify from a liquid containing iron and a small amount of carbon, and previous studies suggest that it provides a good match for the density of the inner core under relevant conditions. Pressure-induced magnetic transitions have been observed in Fe7C3 (Chen et al., 2012). The pressure of the ferromagnetic to paramagnetic transition remains controversial and its effect on equation of state (EoS) is unclear, thus introducing uncertainties in estimating the density of Fe7C3 under inner core pressures. Here we report the lattice parameters and unit cell volume of hexagonal Fe7C3 at 300 K and up to 70 GPa, obtained through synchrotron x-ray diffraction measurements using a diamond anvil cell. The experiments used fine powder of Fe7C3 that was synthesized in the multi-anvil apparatus at the University of Michigan. The sample was embedded in neon pressure medium together with Au powder and ruby spheres as additional pressure markers. We observed significant softening at 5~8 GPa, similar to the reported "invar"-like behavior in Fe-Ni alloy (Dubrovinsky et al., 2001). For comparison, the compression curve of iron in the same loading turned out to be smooth as expected, which confirms that the abnormal behavior in Fe7C3 compression curve is due to its own property change and not an artifact. The new data allow us to establish the equation-of-state (EoS) of Fe7C3 and then estimate the density of Fe7C3 at inner core conditions. References: Chen, B., Gao, L.L., Lavina, B., Dera, P., Alp, E.E., Zhao, J.Y., Li, J., 2012. Magneto-elastic coupling in compressed Fe7C3 supports carbon in Earth's inner core. Geophys Res Lett 39. Dubrovinsky, L., Dubrovinskaia, N., Abrikosov, I.A., Vennstrom, M., Westman, F., Carlson, S., van Schilfgaarde, M., Johansson, B., 2001. Pressure-induced invar effect in Fe-Ni alloys. Phys Rev Lett 86, 4851-4854.

  17. P7C3 neuroprotective chemicals block axonal degeneration and preserve function after traumatic brain injury.

    PubMed

    Yin, Terry C; Britt, Jeremiah K; De Jesús-Cortés, Héctor; Lu, Yuan; Genova, Rachel M; Khan, Michael Z; Voorhees, Jaymie R; Shao, Jianqiang; Katzman, Aaron C; Huntington, Paula J; Wassink, Cassie; McDaniel, Latisha; Newell, Elizabeth A; Dutca, Laura M; Naidoo, Jacinth; Cui, Huxing; Bassuk, Alexander G; Harper, Matthew M; McKnight, Steven L; Ready, Joseph M; Pieper, Andrew A

    2014-09-25

    The P7C3 class of neuroprotective aminopropyl carbazoles has been shown to block neuronal cell death in models of neurodegeneration. We now show that P7C3 molecules additionally preserve axonal integrity after injury, before neuronal cell death occurs, in a rodent model of blast-mediated traumatic brain injury (TBI). This protective quality may be linked to the ability of P7C3 molecules to activate nicotinamide phosphoribosyltransferase, the rate-limiting enzyme in nicotinamide adenine dinucleotide salvage. Initiation of daily treatment with our recently reported lead agent, P7C3-S243, 1 day after blast-mediated TBI blocks axonal degeneration and preserves normal synaptic activity, learning and memory, and motor coordination in mice. We additionally report persistent neurologic deficits and acquisition of an anxiety-like phenotype in untreated animals 8 months after blast exposure. Optimized variants of P7C3 thus offer hope for identifying neuroprotective agents for conditions involving axonal damage, neuronal cell death, or both, such as occurs in TBI. PMID:25220467

  18. P7C3 and an unbiased approach to drug discovery for neurodegenerative diseases

    PubMed Central

    McKnight, Steven L.; Ready, Joseph M.

    2014-01-01

    A novel neuroprotective small molecule was discovered using a target-agnostic in vivo screen in living mice. This aminopropyl carbazole, named P7C3, is orally bioavailable, crosses the blood–brain barrier, and is non-toxic at doses several fold higher than the efficacious dose. The potency and drug-like properties of P7C3 were optimized through a medicinal chemistry campaign, providing analogues for detailed examination. Improved versions, such as (−)-P7C3-S243 and P7C3-A20, displayed neuro-protective properties in rodent models of Parkinson’s disease, amyotrophic lateral sclerosis, traumatic brain injury and age-related cognitive decline. Derivatives appended with immobilizing moieties may reveal the protein targets of the P7C3 class of neuroprotective compounds. Our results indicate that unbiased, in vivo screens might provide starting points for the development of treatments for neurodegenerative diseases as well as tools to study the biology underlying these disorders. PMID:24514864

  19. 76 FR 5066 - Airworthiness Directives; Pratt & Whitney JT8D-7, -7A, -7B, -9, -9A, -11, -15, -15A, -17, -17A...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-28

    ...-14398 (70 FR 73361, December 12, 2005). That AD applies to the specified products. The NPRM published in the Federal Register on August 18, 2010 (75 FR 50942). That NPRM proposed to modify the TLS of the... ``significant rule'' under DOT Regulatory Policies and Procedures (44 FR 11034, February 26, 1979), (3) Will...

  20. 75 FR 50942 - Airworthiness Directives; Pratt & Whitney JT8D-7, -7A, -7B, -9, -9A, -11, -15, -15A, -17, -17A...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-18

    ..., 2000 (65 FR 19477-78). Examining the AD Docket You may examine the AD docket on the Internet at http... AD 2005-25-05, Amendment 39- 14398 (70 FR 73361, December 12, 2005), to require revisions to the TLS.... Is not a ``significant rule'' under the DOT Regulatory Policies and Procedures (44 FR 11034,...

  1. Improving Trichoderma reesei Cel7B thermostability by targeting the weak spots.

    PubMed

    Zhang, Shujun; Wang, Yefei; Song, Xiangfei; Hong, Jingbo; Zhang, Yu; Yao, Lishan

    2014-10-27

    For proteins that denature irreversibly, the denaturation is typically triggered by a partial unfolding, followed by a permanent change (e.g., aggregation). The regions that initiate the partial unfolding are named "weak spots". In this work, a molecular dynamics (MD) simulation and data analysis protocol is developed to identify the weak spots of Trichoderma reesei Cel7B, an important endoglucanase in cellulose hydrolysis, through assigning the local melting temperature (Tmp) to individual residue pairs. To test the predicted weak spots, a total of eight disulfide bonds were designed in these regions and all enhanced the enzyme thermostability. The increased stability, quantified by ΔT50 (which is the T50 difference between the mutant and the wild type enzyme), is negatively correlated with the MD-predicted Tmp, demonstrating the effectiveness of the protocol and highlighting the importance of the weak spots. Strengthening interactions in these regions proves to be a useful strategy in improving the thermostability of Tr. Cel7B. PMID:25286389

  2. Preliminary X-ray analysis of cellobiohydrolase Cel7B from Melanocarpus albomyces

    SciTech Connect

    Parkkinen, Tarja; Koivula, Anu; Vehmaanperä, Jari; Rouvinen, Juha

    2007-09-01

    The crystallization and preliminary X-ray diffraction analysis of cellobiohydrolase from M. albomyces is reported. Cellobiohydrolases are enzymes that cleave off cellobiose units from cellulose chains in a processive manner. Melanocarpus albomyces Cel7B is a thermostable single-module cellobiohydrolase that has relatively low activity on small soluble substrates at room temperature. It belongs to glycoside hydrolase family 7, which includes endo-β-1,4-glucanases and cellobiohydrolases. Cel7B was crystallized using the hanging-drop vapour-diffusion method and streak-seeding. The crystals belonged to space group P2{sub 1}, with unit-cell parameters a = 50.9, b = 94.5, c = 189.8 Å, β = 90.0° and four monomers in the asymmetric unit. Analysis of the intensity statistics showed that the crystals were pseudo-merohedrally twinned, with a twinning fraction of 0.37. X-ray diffraction data were collected at 1.6 Å resolution using synchrotron radiation.

  3. A Bayesian analysis of HAT-P-7b using the EXONEST algorithm

    SciTech Connect

    Placek, Ben; Knuth, Kevin H.

    2015-01-13

    The study of exoplanets (planets orbiting other stars) is revolutionizing the way we view our universe. High-precision photometric data provided by the Kepler Space Telescope (Kepler) enables not only the detection of such planets, but also their characterization. This presents a unique opportunity to apply Bayesian methods to better characterize the multitude of previously confirmed exoplanets. This paper focuses on applying the EXONEST algorithm to characterize the transiting short-period-hot-Jupiter, HAT-P-7b (also referred to as Kepler-2b). EXONEST evaluates a suite of exoplanet photometric models by applying Bayesian Model Selection, which is implemented with the MultiNest algorithm. These models take into account planetary effects, such as reflected light and thermal emissions, as well as the effect of the planetary motion on the host star, such as Doppler beaming, or boosting, of light from the reflex motion of the host star, and photometric variations due to the planet-induced ellipsoidal shape of the host star. By calculating model evidences, one can determine which model best describes the observed data, thus identifying which effects dominate the planetary system. Presented are parameter estimates and model evidences for HAT-P-7b.

  4. Physical state of the deep interior of the CoRoT-7b exoplanet

    NASA Astrophysics Data System (ADS)

    Wagner, Frank W.; Sohl, Frank; Rückriemen, Tina; Rauer, Heike

    2011-11-01

    The present study takes the CoRoT-7b exoplanet as an analogue for massive terrestrial planets to investigate conditions, under which intrinsic magnetic fields could be sustained in liquid cores. We examine the effect of depth-dependent transport parameters (e.g., activation volume of mantle rock) on a planet's thermal structure and the related heat flux across the core mantle boundary. For terrestrial planets more massive than the Earth, our calculations suggest that a substantial part of the lowermost mantle is in a sluggish convective regime, primarily due to pressure effects on viscosity. Hence, we find substantially higher core temperatures than previously reported from parameterized convection models. We also discuss the effect of melting point depression in the presence of impurities (e.g., sulfur) in iron-rich cores and compare corresponding melting relations to the calculated thermal structure. Since impurity effects become less important at the elevated pressure and temperature conditions prevalent in the deep interior of CoRoT-7b, iron-rich cores are likely solid, implying that a self-sustained magnetic field would be absent.

  5. Improving Trichoderma reesei Cel7B thermostability by targeting the weak spots.

    PubMed

    Zhang, Shujun; Wang, Yefei; Song, Xiangfei; Hong, Jingbo; Zhang, Yu; Yao, Lishan

    2014-10-27

    For proteins that denature irreversibly, the denaturation is typically triggered by a partial unfolding, followed by a permanent change (e.g., aggregation). The regions that initiate the partial unfolding are named "weak spots". In this work, a molecular dynamics (MD) simulation and data analysis protocol is developed to identify the weak spots of Trichoderma reesei Cel7B, an important endoglucanase in cellulose hydrolysis, through assigning the local melting temperature (Tmp) to individual residue pairs. To test the predicted weak spots, a total of eight disulfide bonds were designed in these regions and all enhanced the enzyme thermostability. The increased stability, quantified by ΔT50 (which is the T50 difference between the mutant and the wild type enzyme), is negatively correlated with the MD-predicted Tmp, demonstrating the effectiveness of the protocol and highlighting the importance of the weak spots. Strengthening interactions in these regions proves to be a useful strategy in improving the thermostability of Tr. Cel7B.

  6. The role of metal binding and phosphorylation domains in the regulation of cisplatin-induced trafficking of ATP7B.

    PubMed

    Safaei, Roohangiz; Adams, Preston L; Mathews, Ryan A; Manorek, Gerald; Howell, Stephen B

    2013-08-01

    The copper (Cu) exporter ATP7B mediates cellular resistance to cisplatin (cDDP) by increasing drug efflux. ATP7B binds and sequesters cDDP in into secretory vesicles. Upon cDDP exposure ATP7B traffics from the trans-Golgi network (TGN) to the periphery of the cell in a manner that requires the cysteine residues in its metal binding domains (MBD). To elucidate the role of the various domains of ATP7B in its cDDP-induced trafficking we expressed a series of mCherry-tagged variants of ATP7B in HEK293T cells and analyzed their subcellular localization in basal media and after a 1 h exposure to 30 μM cDDP. The wild type ATP7B and a variant in which the cysteines in the CXXC motifs of MBD 1-5 were converted to serines trafficked out of the trans-Golgi (TGN) when exposed to cDDP. Conversion of the cysteines in all 6 of the CXXC motifs to serines, or in only the sixth MBD, rendered ATP7B incapable of trafficking on exposure to cDDP. Truncation of MBD1-5 or MBD1-6 resulted in the loss of TGN localization. Addition of the first 63 amino acids of ATP7B to these variants restored TGN localization to a great extent and enabled the MBD1-5 variant to undergo cDDP-induced trafficking. A variant of ATP7B in which the aspartate 1027 residue in the phosphorylation domain was converted to glutamine localized to the TGN but was incapable of cDDP-induced trafficking. These results demonstrate that the CXXC motif in the sixth MBD and the catalytic activity of ATP7B are required for cDDP-induced trafficking as they are for Cu-induced redistribution of ATP7B; this provides further evidence that cDDP mimics Cu with respect to the molecular mechanisms by they control the subcellular distribution of ATP7B.

  7. The role of metal binding and phosphorylation domains in the regulation of cisplatin-induced trafficking of ATP7B

    PubMed Central

    Safaei, Roohangiz; Adams, Preston L.; Mathews, Ryan A.; Manorek, Gerald; Howell, Stephen B.

    2014-01-01

    The copper (Cu) exporter ATP7B mediates cellular resistance to cisplatin (cDDP) by increasing drug efflux. ATP7B binds and sequesters cDDP in into secretory vesicles. Upon cDDP exposure ATP7B traffics from the trans-Golgi network (TGN) to the periphery of the cell in a manner that requires the cysteine residues in its metal binding domains (MBD). To elucidate the role of the various domains of ATP7B in its cDDP-induced trafficking we expressed a series of mCherry-tagged variants of ATP7B in HEK293T cells and analyzed their subcellular localization in basal media and after a 1 h exposure to 30 μM cDDP. The wild type ATP7B and a variant in which the cysteines in the CXXC motifs of MBD 1-5 were converted to serines trafficked out of the trans-Golgi (TGN) when exposed to cDDP. Conversion of the cysteines in all 6 of the CXXC motifs to serines, or in only the sixth MBD, rendered ATP7B incapable of trafficking on exposure to cDDP. Truncation of MBD1-5 or MBD1-6 resulted in the loss of TGN localization. Addition of the first 63 amino acids of ATP7B to these variants restored TGN localization to a great extent and enabled the MBD1-5 variant to undergo cDDP-induced trafficking. A variant of ATP7B in which the aspartate 1027 residue in the phosphorylation domain was converted to glutamine localized to the TGN but was incapable of cDDP-induced trafficking. These results demonstrate that the CXXC motif in the sixth MBD and the catalytic activity of ATP7B are required for cDDP-induced trafficking as they are for Cu-induced redistribution of ATP7B; this provides further evidence that cDDP mimics Cu with respect to the molecular mechanisms by they control the subcellular distribution of ATP7B. PMID:23803742

  8. Copper transport during lactation in transgenic mice expressing the human ATP7A protein

    PubMed Central

    Llanos, Roxana M.; Michalczyk, Agnes A.; Freestone, David J.; Currie, Scott; Linder, Maria C.; Ackland, M. Leigh; Mercer, Julian F.B.

    2008-01-01

    Both copper transporting ATPases, ATP7A and ATP7B, are expressed in mammary epithelial cells but their role in copper delivery to milk has not been clarified. We investigated the role of ATP7A in delivery of copper to milk using transgenic mice that over-express human ATP7A. In mammary gland of transgenic mice, human ATP7A protein was 10- to 20-fold higher than in control mice, and was localized to the basolateral membrane of mammary epithelial cells in lactating mice. The copper concentration in the mammary gland of transgenic dams and stomach contents of transgenic pups was significantly reduced compared to non-transgenic mice. The mRNA levels of endogenous Atp7a, Atp7b, and Ctr1 copper transporters in the mammary gland were not altered by the expression of the ATP7A transgene, and the protein levels of Atp7b and ceruloplasmin were similar in transgenic and non-transgenic mice. These data suggest that ATP7A plays a role in removing excess copper from the mammary epithelial cells rather than supplying copper to milk. PMID:18515074

  9. Interaction of Drosophila melanogaster prohormone convertase 2 and 7B2. Insect cell-specific processing and secretion.

    PubMed

    Hwang, J R; Siekhaus, D E; Fuller, R S; Taghert, P H; Lindberg, I

    2000-06-01

    The prohormone convertases (PCs) are an evolutionarily ancient group of proteases required for the maturation of neuropeptide and peptide hormone precursors. In Drosophila melanogaster, the homolog of prohormone convertase 2, dPC2 (amontillado), is required for normal hatching behavior, and immunoblotting data indicate that flies express 80- and 75-kDa forms of this protein. Because mouse PC2 (mPC2) requires 7B2, a helper protein for productive maturation, we searched the fly data base for the 7B2 signature motif PPNPCP and identified an expressed sequence tag clone encoding the entire open reading frame for this protein. dPC2 and d7B2 cDNAs were subcloned into expression vectors for transfection into HEK-293 cells; mPC2 and rat 7B2 were used as controls. Although active mPC2 was detected in medium in the presence of either d7B2 or r7B2, dPC2 showed no proteolytic activity upon coexpression of either d7B2 or r7B2. Labeling experiments showed that dPC2 was synthesized but not secreted from HEK-293 cells. However, when dPC2 and either d7B2 or r7B2 were coexpressed in Drosophila S2 cells, abundant immunoreactive dPC2 was secreted into the medium, coincident with the appearance of PC2 activity. Expression and secretion of dPC2 enzyme activity thus appears to require insect cell-specific posttranslational processing events. The significant differences in the cell biology of the insect and mammalian enzymes, with 7B2 absolutely required for secretion of dPC2 and zymogen conversion occurring intracellularly in the case of dPC2 but not mPC2, support the idea that the Drosophila enzyme has specific requirements for maturation and secretion that can be met only in insect cells. PMID:10749852

  10. Using the 7C Framework for Teaching & Learning Health Education & Promotion

    ERIC Educational Resources Information Center

    Becker, Craig M.; Xu, Lei; Chaney, Beth

    2016-01-01

    Health Professionals are needed to address and improve health status. This paper presents a teaching technique that will help students acquire and develop applied health education and promotion skills. This paper introduces a 7C Framework to encourage teachers to use Challenge, Courage, Commitment, Competence, Connection, Contribution, and…

  11. 18 CFR 2.55 - Definition of terms used in section 7(c).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 18 Conservation of Power and Water Resources 1 2013-04-01 2013-04-01 false Definition of terms used in section 7(c). 2.55 Section 2.55 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY GENERAL RULES GENERAL POLICY AND INTERPRETATIONS Statements...

  12. 18 CFR 2.55 - Definition of terms used in section 7(c).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 18 Conservation of Power and Water Resources 1 2010-04-01 2010-04-01 false Definition of terms used in section 7(c). 2.55 Section 2.55 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY GENERAL RULES GENERAL POLICY AND INTERPRETATIONS Statements...

  13. 18 CFR 2.55 - Definition of terms used in section 7(c).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 18 Conservation of Power and Water Resources 1 2012-04-01 2012-04-01 false Definition of terms used in section 7(c). 2.55 Section 2.55 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY GENERAL RULES GENERAL POLICY AND INTERPRETATIONS Statements...

  14. Functional analysis and drug response to zinc and D-penicillamine in stable ATP7B mutant hepatic cell lines

    PubMed Central

    Chandhok, Gursimran; Horvath, Judit; Aggarwal, Annu; Bhatt, Mohit; Zibert, Andree; Schmidt, Hartmut HJ

    2016-01-01

    AIM: To study the effect of anti-copper treatment for survival of hepatic cells expressing different ATP7B mutations in cell culture. METHODS: The most common Wilson disease (WD) mutations p.H1069Q, p.R778L and p.C271*, found in the ATP7B gene encoding a liver copper transporter, were studied. The mutations represent major genotypes of the United States and Europe, China, and India, respectively. A human hepatoma cell line previously established to carry a knockout of ATP7B was used to stably express WD mutants. mRNA and protein expression of mutant ATP7B, survival of cells, apoptosis, and protein trafficking were determined. RESULTS: Low temperature increased ATP7B protein expression in several mutants. Intracellular ATP7B localization was significantly impaired in the mutants. Mutants were classified as high, moderate, and no survival based on their viability on exposure to toxic copper. Survival of mutant p.H1069Q and to a lesser extent p.C271* improved by D-penicillamine (DPA) treatment, while mutant p.R778L showed a pronounced response to zinc (Zn) treatment. Overall, DPA treatment resulted in higher cell survival as compared to Zn treatment; however, only combined Zn + DPA treatment fully restored cell viability. CONCLUSION: The data indicate that the basic impact of a genotype might be characterized by analysis of mutant hepatic cell lines. PMID:27122662

  15. Age-related change in plasma concentration of 7B2(a novel pituitary polypeptide) in normal humans

    SciTech Connect

    Natori, S.; Iguchi, H.; Nawata, H.; Kato, K.; Ibayashi, H.; Chretian, M.

    1987-08-24

    Using a specific radioimmunoassay, the authors measured concentrations of plasma 7B2(a novel pituitary polypeptide) immunoreactivity(7B2-IR) in normal human subjects, patients with chronic renal failure and those with liver cirrhosis. Mean(+/-SEM) values of plasma 7B2-IR in normal healthy men and women were 55.8 +/- 1.2 pg/ml (n=266) and 56.1 +/- 0.9 pg/ml (n = 408), respectively. The elevation of plasma 7B2-IR showed a relationship with age of the subjects, in both men(r=0.39, t = 6.86, p < 0.001) and women (r=0.35, t=7.44, p < 0.001). Plasma 7B2-IR concentrations were elevated in patients with chronic renal failure (536 +/- 45 pg/ml, Mean +/- SEM, n = 10) as well as those in liver cirrhosis (95 +/- 10 pg/ml, Mean +/- SEM, n = 15) compared to values in normal subjects, suggesting that 7B2 is mainly eliminated through the kidney and is partly metabolized in the liver. 10 references, 4 figures.

  16. HATS-7b: A Hot Super Neptune Transiting a Quiet K Dwarf Star

    NASA Astrophysics Data System (ADS)

    Bakos, G. Á.; Penev, K.; Bayliss, D.; Hartman, J. D.; Zhou, G.; Brahm, R.; Mancini, L.; de Val-Borro, M.; Bhatti, W.; Jordán, A.; Rabus, M.; Espinoza, N.; Csubry, Z.; Howard, A. W.; Fulton, B. J.; Buchhave, L. A.; Ciceri, S.; Henning, T.; Schmidt, B.; Isaacson, H.; Noyes, R. W.; Marcy, G. W.; Suc, V.; Howe, A. R.; Burrows, A. S.; Lázár, J.; Papp, I.; Sári, P.

    2015-11-01

    We report the discovery by the HATSouth network of HATS-7b, a transiting Super-Neptune with a mass of 0.120 ± 0.012 {M}{{J}}, a radius of {0.563}-0.034+0.046 {R}{{J}}, and an orbital period of 3.1853 days. The host star is a moderately bright (V=13.340\\+/- 0.010 mag, {K}S=10.976\\+/- 0.026 mag) K dwarf star with a mass of 0.849 ± 0.027 {M}ȯ , a radius of {0.815}-0.035+0.049 {R}ȯ , and a metallicity of [{Fe}/{{H}}] =+0.250\\+/- 0.080. The star is photometrically quiet to within the precision of the HATSouth measurements, has low RV jitter, and shows no evidence for chromospheric activity in its spectrum. HATS-7b is the second smallest radius planet discovered by a wide-field ground-based transit survey, and one of only a handful of Neptune-size planets with mass and radius determined to 10% precision. Theoretical modeling of HATS-7b yields a hydrogen–helium fraction of 18 ± 4% (rock-iron core and H2–He envelope), or 9 ± 4% (ice core and H2–He envelope), i.e., it has a composition broadly similar to that of Uranus and Neptune, and very different from that of Saturn, which has 75% of its mass in H2–He. Based on a sample of transiting exoplanets with accurately (<20%) determined parameters, we establish approximate power-law relations for the envelopes of the mass–density distribution of exoplanets. HATS-7b, which, together with the recently discovered HATS-8b, is one of the first two transiting super-Neptunes discovered in the Southern sky, is a prime target for additional follow-up observations with Southern hemisphere facilities to characterize the atmospheres of Super-Neptunes (which we define as objects with mass greater than that of Neptune, and smaller than halfway between that of Neptune and Saturn, i.e., 0.054 {M}{{J}}\\lt {M}{{p}}\\lt 0.18 {M}{{J}}). The HATSouth network is operated by a collaboration consisting of Princeton University (PU), the Max Planck Institute für Astronomie (MPIA), the Australian National University (ANU), and the

  17. HATS-7b: A Hot Super Neptune Transiting a Quiet K Dwarf Star

    NASA Astrophysics Data System (ADS)

    Bakos, G. Á.; Penev, K.; Bayliss, D.; Hartman, J. D.; Zhou, G.; Brahm, R.; Mancini, L.; de Val-Borro, M.; Bhatti, W.; Jordán, A.; Rabus, M.; Espinoza, N.; Csubry, Z.; Howard, A. W.; Fulton, B. J.; Buchhave, L. A.; Ciceri, S.; Henning, T.; Schmidt, B.; Isaacson, H.; Noyes, R. W.; Marcy, G. W.; Suc, V.; Howe, A. R.; Burrows, A. S.; Lázár, J.; Papp, I.; Sári, P.

    2015-11-01

    We report the discovery by the HATSouth network of HATS-7b, a transiting Super-Neptune with a mass of 0.120 ± 0.012 {M}{{J}}, a radius of {0.563}-0.034+0.046 {R}{{J}}, and an orbital period of 3.1853 days. The host star is a moderately bright (V=13.340\\+/- 0.010 mag, {K}S=10.976\\+/- 0.026 mag) K dwarf star with a mass of 0.849 ± 0.027 {M}⊙ , a radius of {0.815}-0.035+0.049 {R}⊙ , and a metallicity of [{Fe}/{{H}}] =+0.250\\+/- 0.080. The star is photometrically quiet to within the precision of the HATSouth measurements, has low RV jitter, and shows no evidence for chromospheric activity in its spectrum. HATS-7b is the second smallest radius planet discovered by a wide-field ground-based transit survey, and one of only a handful of Neptune-size planets with mass and radius determined to 10% precision. Theoretical modeling of HATS-7b yields a hydrogen-helium fraction of 18 ± 4% (rock-iron core and H2-He envelope), or 9 ± 4% (ice core and H2-He envelope), i.e., it has a composition broadly similar to that of Uranus and Neptune, and very different from that of Saturn, which has 75% of its mass in H2-He. Based on a sample of transiting exoplanets with accurately (<20%) determined parameters, we establish approximate power-law relations for the envelopes of the mass-density distribution of exoplanets. HATS-7b, which, together with the recently discovered HATS-8b, is one of the first two transiting super-Neptunes discovered in the Southern sky, is a prime target for additional follow-up observations with Southern hemisphere facilities to characterize the atmospheres of Super-Neptunes (which we define as objects with mass greater than that of Neptune, and smaller than halfway between that of Neptune and Saturn, i.e., 0.054 {M}{{J}}\\lt {M}{{p}}\\lt 0.18 {M}{{J}}). The HATSouth network is operated by a collaboration consisting of Princeton University (PU), the Max Planck Institute für Astronomie (MPIA), the Australian National University (ANU), and the Pontificia

  18. The KDM2B- let-7b -EZH2 axis in myelodysplastic syndromes as a target for combined epigenetic therapy.

    PubMed

    Karoopongse, Ekapun; Yeung, Cecilia; Byon, John; Ramakrishnan, Aravind; Holman, Zaneta J; Jiang, Peter Y Z; Yu, Qiang; Deeg, H Joachim; Marcondes, A Mario

    2014-01-01

    Both DNA and histone methylation are dysregulated in the myelodysplastic syndromes (MDS). Based on preliminary data we hypothesized that dysregulated interactions of KDM2B, let-7b and EZH2 signals lead to an aberrant epigenetic landscape. Gene expression in CD34+ cells from MDS marrows was analyzed by NanoString miR array and validated by real-time polymerase chain reaction (PCR). The functions of KDM2B, let-7b and EZH2 were characterized in myeloid cell lines and in primary MDS cells. Let-7b levels were significantly higher, and KDM2B and EZH2 expression was lower in primary CD34+ MDS marrow cells (n = 44) than in healthy controls (n = 21; p<0.013, and p<0.0001, respectively). Overexpression of let-7b reduced EZH2 and KDM2B protein levels, and decreased cells in S-phase while increasing G0/G1 cells (p = 0.0005), accompanied by decreased H3K27me3 and cyclin D1. Silencing of KDM2B increased let-7b expression. Treatment with the cyclopentanyl analog of 3-deazaadenosine, DZNep, combined with the DNA hypomethylating agent 5-azacitidine, decreased levels of EZH2, suppressed methylation of di- and tri-methylated H3K27, and increased p16 expression, associated with cell proliferation. Thus, KDM2B, via let-7b/EZH2, promotes transcriptional repression. DZNep bypassed the inhibitory KDM2B/let-7b/EZH2 axis by preventing H3K27 methylation and reducing cell proliferation. DZNep might be able to enhance the therapeutic effects of DNA hypomethylating agents such as 5-azacitidine, currently considered standard therapy for patients with MDS. PMID:25225797

  19. General access to taiwaniaquinoids based on a hypothetical abietane C7-C8 cleavage biogenetic pathway.

    PubMed

    Tapia, Rubén; Guardia, Juan J; Alvarez, Esteban; Haidöur, Ali; Ramos, Jose M; Alvarez-Manzaneda, Ramón; Chahboun, Rachid; Alvarez-Manzaneda, Enrique

    2012-01-01

    A new strategy for synthesizing taiwaniaquinoids, a group of terpenoids with an unusual rearranged 5(6→7) or 6-nor-5(6→7)abeo-abietane skeleton, which exhibit promising biological activities, is reported. The procedure, based on the cleavage of the C7-C8 double bond of abietane diterpenes, is the only one yet reported for synthesizing C(20) taiwaniaquinoids bearing a carbon function on the cyclopentane B ring; it is also applicable to the synthesis of the wide variety of existing taiwaniaquinoids. Utilizing this, (-)-taiwaniaquinone A, F, G, and H, (-)-taiwaniaquinol B, and (-)-dichroanone have been synthesized from (+)-abietic acid. The versatility of this strategy allows us to propose the abietane C7-C8 cleavage as a possible biosynthetic pathway to this type of rearranged diterpenes; this proposal seems to be supported by phytochemical evidence.

  20. Biosynthesis of poly(3-hydroxybutyrateco-3-hydroxy-4-methylvalerate) by Strain Azotobacter chroococcum 7B

    PubMed Central

    Bonartsev, A.P.; Bonartseva, G. A.; Myshkina, V. L.; Voinova, V. V.; Mahina, T. K.; Zharkova, I. I.; Yakovlev, S. G.; Zernov, A. L.; Ivanova, E. V.; Akoulina, E. A.; Kuznetsova, E. S.; Zhuikov, V. A.; Alekseeva, S. G.; Podgorskii, V. V.; Bessonov, I. V.; Kopitsyna, M. N.; Morozov, A. S.; Milanovskiy, E. Y.; Tyugay, Z. N.; Bykova, G. S.; Kirpichnikov, M. P.; Shaitan, K. V.

    2016-01-01

    Production of novel polyhydroxyalkanoates (PHAs), biodegradable polymers for biomedical applications, and biomaterials based on them is a promising trend in modern bioengineering. We studied the ability of an effective strain-producer Azotobacter chroococcum 7B to synthesize not only poly(3-hydroxybutyrate) homopolymer (PHB) and its main copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), but also a novel copolymer, poly(3-hydroxybutyrate-co-3-hydroxy-4-methylvalerate) (PHB4MV). For the biosynthesis of PHB copolymers, we used carboxylic acids as additional carbon sources and monomer precursors in the chain of synthesized copolymers. The main parameters of these polymers’ biosynthesis were determined: strain-producer biomass yield, polymer yield, molecular weight and monomer composition of the synthesized polymers, as well as the morphology of A. chroococcum 7B bacterial cells. The physico-chemical properties of the polymers were studied using nuclear magnetic resonance spectroscopy (NMR), differential scanning calorimetry (DSC), contact angle test, and other methods. In vitro biocompatibility of the obtained polymers was investigated using stromal cells isolated from the bone marrow of rats with the XTT cell viability test. The synthesis of the novel copolymer PHB4MV and its chemical composition were demonstrated by NMR spectroscopy: the addition of 4-methylvaleric acid to the culture medium resulted in incorporation of 3-hydroxy-4-methylvalerate (3H4MV) monomers into the PHB polymer chain (0.6 mol%). Despite the low molar content of 3H4MV in the obtained copolymer, its physico-chemical properties were significantly different from those of the PHB homopolymer: it has lower crystallinity and a higher contact angle, i.e. the physico-chemical properties of the PHB4MV copolymer containing only 0.6 mol% of 3H4MV corresponded to a PHBV copolymer with a molar content ranging from 2.5% to 7.8%. In vitro biocompatibility of the obtained PHB4MV copolymer

  1. DNA Methylation and Transcriptomic Changes in Response to Different Lights and Stresses in 7B-1 Male-Sterile Tomato

    PubMed Central

    Omidvar, Vahid; Fellner, Martin

    2015-01-01

    We reported earlier that 7B-1 mutant in tomato (Solanum lycopersicum L., cv. Rutgers), an ABA overproducer, is defective in blue light (B) signaling leading to B-specific resistance to abiotic and biotic stresses. Using a methylation-sensitive amplified polymorphism (MSAP) assay, a number of genes were identified, which were differentially methylated between 7B-1 and its wild type (WT) seedlings in white (W), blue (B), red (R) lights and dark (D) or in response to exogenous ABA and mannitol-induced stresses. The genomic methylation level was almost similar in different lights between 7B-1 and WT seedlings, while significant differences were observed in response to stresses in D, but not B. Using a cDNA-AFLP assay, several transcripts were identified, which were differentially regulated between 7B-1 and WT by B or D or in response to stresses. Blue light receptors cryptochrome 1 and 2 (CRY1 and CRY2) and phototropin 1 and 2 (PHOT1 and PHOT2) were not affected by the 7B-1 mutation at the transcriptional level, instead the mutation had likely affected downstream components of the light signaling pathway. 5-azacytidine (5-azaC) induced DNA hypomethylation, inhibited stem elongation and differentially regulated the expression of a number of genes in 7B-1. In addition, it was shown that mir167 and mir390 were tightly linked to auxin signaling pathway in 5-azaC-treated 7B-1 seedlings via the regulation of auxin-response factor (ARF) transcripts. Our data showed that DNA methylation remodeling is an active epigenetic response to different lights and stresses in 7B-1 and WT, and highlighted the differences in epigenetic and transcriptional regulation of light and stress responses between 7B-1 and WT. Furthermore, it shed lights on the crosstalk between DNA hypomethylation and miRNA regulation of ARFs expression. This information could also be used as a benchmark for future studies of male-sterility in other crops. PMID:25849771

  2. Fragmentation of mitochondrial cardiolipin by copper ions in the Atp7b-/- mouse model of Wilson's disease.

    PubMed

    Yurkova, Irina L; Arnhold, Juergen; Fitzl, Guenther; Huster, Dominik

    2011-07-01

    Cellular copper overload as found in Wilson's disease may disturb mitochondrial function and integrity. Atp7b(-/-) mice accumulate copper in the liver and serve as an animal model for this inherited disease. The molecular mechanism of copper toxicity in hepatocytes is poorly understood. Total mitochondrial lipids from liver of wild-type mice were subjected to oxidative stress by the Cu(2+)/H(2)O(2)/ascorbate system. Phosphatidic acid (PA) and phosphatidylhydroxyacetone (PHA) were detected as cardiolipin fragmentation products by thin-layer chromatography combined with MALDI-TOF mass spectrometry in oxidized samples, but not in unperturbed ones. The formation of PA and PHA in copper-treated model membrane correlated well with the decrease of cardiolipin. Mitochondrial lipids from Atp7b(-/-) mice of different age were analyzed for the presence of PA. While 32-weeks old wild-type (control) and Atp7b(-/-) mice did not show any PA, there was a steady increase in the amount of this lipid in Atp7b(-/-) mice in contrast to control with increasing age. Hepatocytes from elder Atp7b(-/-)mice contained morphologically changed mitochondria unlike cells from wild-type animals of the same age. We concluded that free-radical fragmentation of cardiolipin with the formation of PA is a likely mechanism that damages mitochondria under conditions of oxidative stress due to copper overload. Our findings are relevant for better understanding of molecular mechanisms for liver damage found in Wilson's disease.

  3. Mechanical properties and chemical bonding characteristics of Cr7C3 type multicomponent carbides

    NASA Astrophysics Data System (ADS)

    Xiao, B.; Feng, J.; Zhou, C. T.; Jiang, Y. H.; Zhou, R.

    2011-01-01

    The first principles calculations based on density functional theory are performed to investigate the mechanical properties and chemical bonding features of several Cr7C3 type multicomponent carbides (Fe16Cr12C12, Fe12Cr12W4C12, Fe12Cr12Mo4C12, Fe12Cr12W4C8B4, and Fe12Cr12Mo4C8B4). The full set elastic constants are calculated using stress-strain method. The Voigt-Reuss-Hill approximation is used to evaluate the mechanical moduli. The mechanical anisotropy is characterized by calculating several different anisotropic indexes and factors, such as universal anisotropic index (AU), shear anisotropic factors (A1, A2, and A3), and percent anisotropy (AB and AG). The surface constructions of bulk and Young's moduli are illustrated to indicate the mechanical anisotropy. The obtained results indicate that the presence of B could deteriorate the mechanical moduli of (Fe,Cr)7C3 phase in Cr cast iron slightly. On the other hand, the chemical stability of (Fe,Cr)7C3 carbides is improved due to chemical bonding modifications through doping with B. These conclusions are in agreement with experimental results.

  4. Evaluating endoglucanase Cel7B-lignin interaction mechanisms and kinetics using quartz crystal microgravimetry.

    PubMed

    Pfeiffer, Katherine A; Sorek, Hagit; Roche, Christine M; Strobel, Kathryn L; Blanch, Harvey W; Clark, Douglas S

    2015-11-01

    The kinetics and mechanisms of protein interactions with solid surfaces are important to fields as diverse as industrial biocatalysis, biomedical engineering, food science, and cell biology. The nonproductive adsorption of cellulase enzymes to lignin, a plant cell wall polymer, reduces their effectiveness in saccharifying biomass. Cellulase has been shown to interact with lignin, but the heterogeneity of lignin surfaces, challenges in measuring irreversible components of these interactions, and fast adsorption rates make quantifying the reaction kinetics difficult. This work employs quartz crystal microgravimetry with dissipation monitoring (QCM-D) for real-time measurement of adsorbed mass on a flat lignin surface. We have developed a method for casting homogeneous lignin films that are chemically similar to lignin found in pretreated biomass, and used QCM-D to compare three models of reversible-irreversible binding behavior: a single-site transition model, a transition model with changing adsorbate footprint, and a two-site transition model. Of the three models tested, the two-site transition model provides the only kinetic mechanism able to describe the behavior of Cel7B binding to lignin. While the direct implications of lignin-cellulase interactions may be limited to biomass deconstruction for renewable energy and green chemistry, the analytical and experimental methods demonstrated in this work are relevant to any system in which the kinetics and reaction mechanism of reversible and irreversible protein adsorption at a solid-liquid interface are important. PMID:25994114

  5. Characterization of the Dielectric Constant in the Trichoderma reesei Cel7B Active Site.

    PubMed

    Song, Xiangfei; Wang, Yefei; Zhang, Shujun; Yan, Shihai; Li, Tong; Yao, Lishan

    2015-07-27

    An attempt is made to evaluate the dielectric constant of the Trichoderma reesei Cel7B active site. Through kinetic measurements, the pKa value of the catalytic acid E201 is determined. Mutations (away from E201) with net charge changes are introduced to perturb the E201 pKa. It is shown that the mutation with a +1 charge change (including G225R, G230R, and A335R) decreases the pKa of E201, whereas the mutation with a -1 charge change (including Q149E, A222D, G225D, and G230D) increases the pKa. This effect is consistent with the electrostatic interaction between the changed charge and the E201 side chain. The fitting of the experimental data yields an apparent dielectric constant of 25-80. Molecular dynamics simulations with explicit water molecules indicate that the high solvent accessibility of the active site contributes largely to the high dielectric constant. ONIOM calculations show that high dielectric constant benefits the catalysis through decreasing the energy of the transition state relative to that of the enzyme substrate complex. PMID:26114648

  6. A mechanistic study of Trichoderma reesei Cel7B catalyzed glycosidic bond cleavage.

    PubMed

    Zhang, Yu; Yan, Shihai; Yao, Lishan

    2013-07-25

    An ONIOM study is performed to illustrate the mechanism of Trichoderma reesei Cel7B catalyzed p-nitrophenyl lactoside hydrolysis. In both the glycosylation and deglycosylation steps, the reaction proceeds in a concerted way, meaning the nucleophilic attack and the glycosidic bond cleavage occur simultaneously. The glycosylation step is rate limiting with a barrier of 18.9 kcal/mol, comparable to the experimental value derived from the kcat measured in this work. The function of four residues R108, Y146, Y170, and D172, which form a hydrogen-bond network involving the substrate, is studied by conservative mutations. The mutants, including R108K, Y146F, Y170F, and D172N, decrease the enzyme activity by about 150-8000-fold. Molecular dynamics simulations show that the mutations disrupt the hydrogen-bond network, cause the substrate to deviate from active binding and hinder either the proton transfer from E201 to O4(+1) or the nucleophilic attack from E196 to C1(-1). PMID:23822607

  7. An investigation of higher-order effects in modeling exoplanet HAT-P-7b

    NASA Astrophysics Data System (ADS)

    Kurth, Susan Anne

    In its search for Earth-like planets, NASA's Kepler mission observed over 200,000 stars. Among these systems were the class of planets known as the "hot Jupiters." These are giant gaseous planets with periods less than 10 days. Here I present an analysis of the Kepler observations of the exoplanet HAT-P-7b, a hot Jupiter with an orbital period of 2.2 days, a mass of 1.8 MJ, and a radius of 1.5 RJ. Due to its very close proximity to its host star, the planet has a day-side temperature of 2800 K and a night-side temperature of 1950 K. The tight orbit also causes planetary reflection, as well as higher-order effects in the system such as ellipsoidal variations, Doppler beaming, and gravity darkening. My thesis explores these effects on the Kepler HAT-P-7 light curve using the state of the art Eclipsing Light Curve (ELC) code. The "short cadence" data from Kepler contain 2 million measurements, which I phase folded and binned to get robust uncertainties, resulting in a final data set with 2,704 points. Because of the exquisitely high precision of the data (few ppm level), the physical effects mentioned above need to be accounted for. Including these effects enables me to accurately solve for the system parameters.

  8. The Terpolymer Produced by Azotobacter Chroococcum 7B: Effect of Surface Properties on Cell Attachment

    PubMed Central

    Bonartsev, Anton; Yakovlev, Sergey; Boskhomdzhiev, Arasha; Zharkova, Irina; Bagrov, Dmitrii; Myshkina, Vera; Mahina, Tatiana; Kharitonova, Elena; Samsonova, Olga; Zernov, Anton; Zhuikov, Vsevolod; Efremov, Yurii; Voinova, Vera; Bonartseva, Garina; Shaitan, Konstantin

    2013-01-01

    The copolymerization of poly(3-hydroxybutyrate) (PHB) is a promising trend in bioengineering to improve biomedical properties, e.g. biocompatibility, of this biodegradable polymer. We used strain Azotobacter chroococcum 7B, an effective producer of PHB, for biosynthesis of not only homopolymer and its main copolymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHB-HV), but also novel terpolymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate)-poly(ethylene glycol) (PHB-HV-PEG), using sucrose as the primary carbon source and valeric acid and poly(ethylene glycol) 300 (PEG 300) as additional carbon sources. The chemical structure of PHB-HV-PEG was confirmed by 1H nuclear-magnetic resonance analysis. The physico-chemical properties (molecular weight, crystallinity, hydrophilicity, surface energy) of produced biopolymer, the protein adsorption to the terpolymer, and cell growth on biopolymer films were studied. Despite of low EG-monomers content in bacterial-origin PHB-HV-PEG polymer, the terpolymer demonstrated significant improvement in biocompatibility in vitro in contrast to PHB and PHB-HV polymers, which may be coupled with increased protein adsorption, hydrophilicity and surface roughness of PEG-containing copolymer. PMID:23468935

  9. Characterization of the Dielectric Constant in the Trichoderma reesei Cel7B Active Site.

    PubMed

    Song, Xiangfei; Wang, Yefei; Zhang, Shujun; Yan, Shihai; Li, Tong; Yao, Lishan

    2015-07-27

    An attempt is made to evaluate the dielectric constant of the Trichoderma reesei Cel7B active site. Through kinetic measurements, the pKa value of the catalytic acid E201 is determined. Mutations (away from E201) with net charge changes are introduced to perturb the E201 pKa. It is shown that the mutation with a +1 charge change (including G225R, G230R, and A335R) decreases the pKa of E201, whereas the mutation with a -1 charge change (including Q149E, A222D, G225D, and G230D) increases the pKa. This effect is consistent with the electrostatic interaction between the changed charge and the E201 side chain. The fitting of the experimental data yields an apparent dielectric constant of 25-80. Molecular dynamics simulations with explicit water molecules indicate that the high solvent accessibility of the active site contributes largely to the high dielectric constant. ONIOM calculations show that high dielectric constant benefits the catalysis through decreasing the energy of the transition state relative to that of the enzyme substrate complex.

  10. A mechanistic study of Trichoderma reesei Cel7B catalyzed glycosidic bond cleavage.

    PubMed

    Zhang, Yu; Yan, Shihai; Yao, Lishan

    2013-07-25

    An ONIOM study is performed to illustrate the mechanism of Trichoderma reesei Cel7B catalyzed p-nitrophenyl lactoside hydrolysis. In both the glycosylation and deglycosylation steps, the reaction proceeds in a concerted way, meaning the nucleophilic attack and the glycosidic bond cleavage occur simultaneously. The glycosylation step is rate limiting with a barrier of 18.9 kcal/mol, comparable to the experimental value derived from the kcat measured in this work. The function of four residues R108, Y146, Y170, and D172, which form a hydrogen-bond network involving the substrate, is studied by conservative mutations. The mutants, including R108K, Y146F, Y170F, and D172N, decrease the enzyme activity by about 150-8000-fold. Molecular dynamics simulations show that the mutations disrupt the hydrogen-bond network, cause the substrate to deviate from active binding and hinder either the proton transfer from E201 to O4(+1) or the nucleophilic attack from E196 to C1(-1).

  11. Evaluating endoglucanase Cel7B-lignin interaction mechanisms and kinetics using quartz crystal microgravimetry.

    PubMed

    Pfeiffer, Katherine A; Sorek, Hagit; Roche, Christine M; Strobel, Kathryn L; Blanch, Harvey W; Clark, Douglas S

    2015-11-01

    The kinetics and mechanisms of protein interactions with solid surfaces are important to fields as diverse as industrial biocatalysis, biomedical engineering, food science, and cell biology. The nonproductive adsorption of cellulase enzymes to lignin, a plant cell wall polymer, reduces their effectiveness in saccharifying biomass. Cellulase has been shown to interact with lignin, but the heterogeneity of lignin surfaces, challenges in measuring irreversible components of these interactions, and fast adsorption rates make quantifying the reaction kinetics difficult. This work employs quartz crystal microgravimetry with dissipation monitoring (QCM-D) for real-time measurement of adsorbed mass on a flat lignin surface. We have developed a method for casting homogeneous lignin films that are chemically similar to lignin found in pretreated biomass, and used QCM-D to compare three models of reversible-irreversible binding behavior: a single-site transition model, a transition model with changing adsorbate footprint, and a two-site transition model. Of the three models tested, the two-site transition model provides the only kinetic mechanism able to describe the behavior of Cel7B binding to lignin. While the direct implications of lignin-cellulase interactions may be limited to biomass deconstruction for renewable energy and green chemistry, the analytical and experimental methods demonstrated in this work are relevant to any system in which the kinetics and reaction mechanism of reversible and irreversible protein adsorption at a solid-liquid interface are important.

  12. Identification of myopia-associated WNT7B polymorphisms provides insights into the mechanism underlying the development of myopia.

    PubMed

    Miyake, Masahiro; Yamashiro, Kenji; Tabara, Yasuharu; Suda, Kenji; Morooka, Satoshi; Nakanishi, Hideo; Khor, Chiea-Chuen; Chen, Peng; Qiao, Fan; Nakata, Isao; Akagi-Kurashige, Yumiko; Gotoh, Norimoto; Tsujikawa, Akitaka; Meguro, Akira; Kusuhara, Sentaro; Polasek, Ozen; Hayward, Caroline; Wright, Alan F; Campbell, Harry; Richardson, Andrea J; Schache, Maria; Takeuchi, Masaki; Mackey, David A; Hewitt, Alex W; Cuellar, Gabriel; Shi, Yi; Huang, Luling; Yang, Zhenglin; Leung, Kim Hung; Kao, Patrick Y P; Yap, Maurice K H; Yip, Shea Ping; Moriyama, Muka; Ohno-Matsui, Kyoko; Mizuki, Nobuhisa; MacGregor, Stuart; Vitart, Veronique; Aung, Tin; Saw, Seang-Mei; Tai, E-Shyong; Wong, Tien Yin; Cheng, Ching-Yu; Baird, Paul N; Yamada, Ryo; Matsuda, Fumihiko; Yoshimura, Nagahisa

    2015-03-31

    Myopia can cause severe visual impairment. Here, we report a two-stage genome-wide association study for three myopia-related traits in 9,804 Japanese individuals, which was extended with trans-ethnic replication in 2,674 Chinese and 2,690 Caucasian individuals. We identify WNT7B as a novel susceptibility gene for axial length (rs10453441, Pmeta=3.9 × 10(-13)) and corneal curvature (Pmeta=2.9 × 10(-40)) and confirm the previously reported association between GJD2 and myopia. WNT7B significantly associates with extreme myopia in a case-control study with 1,478 Asian patients and 4,689 controls (odds ratio (OR)meta=1.13, Pmeta=0.011). We also find in a mouse model of myopia downregulation of WNT7B expression in the cornea and upregulation in the retina, suggesting its possible role in the development of myopia.

  13. Effect of Carbide Ceramic Zone on Wear Resistance of the (Fe,Cr)7C3/Fe Surface Gradient Composite

    NASA Astrophysics Data System (ADS)

    Ye, Fangxia; Xu, Yunhua; Hojamberdiev, Mirabbos; Lai, Yujun; Wang, Chong; Wang, Xin

    2015-08-01

    In this work, we report on the influence of microstructure and mechanical properties of the (Fe,Cr)7C3 ceramic zone on wear resistance of the (Fe,Cr)7C3/Fe surface gradient composite fabricated by in situ synthesis method followed by a post-heat treatment at 1100 °C for 20 h in argon atmosphere. The phase composition, microstructure, nanoindentation hardness, elastic modulus, fracture toughness, and relative wear resistance of the (Fe,Cr)7C3/Fe surface gradient composite were investigated by means of x-ray diffraction, scanning electron microscopy, nanoindentation tester, and wear resistance testing instrument, respectively. The XRD results showed that (Fe,Cr)7C3 is the predominant crystalline phases in the fabricated composite. The volume fraction of the (Fe,Cr)7C3 particulates formed has a gradient distribution from the surface to the iron matrix, and the microstructure also changes significantly. The (Fe,Cr)7C3 bulk ceramic zone with the volume fraction of about 100% and the (Fe,Cr)7C3 dense ceramic zone with the volume fraction of about 90% were synthesized on the upper surface of the (Fe,Cr)7C3/Fe surface gradient composite, respectively. The average nanoindentation hardness and elastic modulus of the (Fe,Cr)7C3 bulk ceramic zone of the composite were determined to be 12.711 and 256.054 GPa, respectively. The fracture toughness of the (Fe,Cr)7C3 bulk ceramic zone is in the range of 2.06-4.19 MPa m1/2, and its relative wear resistance is about 56 times higher than that of the iron matrix. The (Fe,Cr)7C3 dense ceramic zone with rod-like, secondary (Fe,Cr)7C3 particulates was formed at the bottom of the (Fe,Cr)7C3 bulk ceramic zone. Rod-like, secondary (Fe,Cr)7C3 particulates are dense and grew in the direction of the iron substrate, providing higher wear resistance to the composite. The wear mechanisms of the (Fe,Cr)7C3 bulk and dense ceramic zones are considered to be microcutting, microcracking, and spalling pit.

  14. 29 CFR 516.21 - Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section 7(b...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 3 2012-07-01 2012-07-01 false Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section 7(b)(3) of the Act. 516.21 Section 516.21 Labor Regulations Relating to Labor....21 Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section...

  15. 29 CFR 516.21 - Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section 7(b...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 3 2013-07-01 2013-07-01 false Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section 7(b)(3) of the Act. 516.21 Section 516.21 Labor Regulations Relating to Labor....21 Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section...

  16. 29 CFR 516.21 - Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section 7(b...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 3 2014-07-01 2014-07-01 false Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section 7(b)(3) of the Act. 516.21 Section 516.21 Labor Regulations Relating to Labor....21 Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section...

  17. 29 CFR 516.21 - Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section 7(b...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section 7(b)(3) of the Act. 516.21 Section 516.21 Labor Regulations Relating to Labor....21 Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section...

  18. 29 CFR 516.21 - Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section 7(b...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 3 2011-07-01 2011-07-01 false Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section 7(b)(3) of the Act. 516.21 Section 516.21 Labor Regulations Relating to Labor....21 Bulk petroleum employees partially exempt from overtime pay requirements pursuant to section...

  19. Multi-allele genotyping platform for the simultaneous detection of mutations in the Wilson disease related ATP7B gene.

    PubMed

    Amvrosiadou, Maria; Petropoulou, Margarita; Poulou, Myrto; Tzetis, Maria; Kanavakis, Emmanuel; Christopoulos, Theodore K; Ioannou, Penelope C

    2015-12-01

    Wilson's disease is an inherited disorder of copper transport in the hepatocytes with a wide range of genotype and phenotype characteristics. Mutations in the ATP7B gene are responsible for the disease. Approximately, over 500 mutations in the ATP7B gene have been described to date. We report a method for the simultaneous detection of the ten most common ATP7B gene mutations in Greek patients. The method comprises 3 simple steps: (i) multiplex PCR amplification of fragments in the ATP7B gene flanking the mutations (ii) multiplex primer extension reaction of the unpurified amplification products using allele-specific primers and (iii) visual detection of the primer extension reaction products within minutes by means of dry-reagent multi-allele dipstick assay using anti-biotin conjugated gold nanoparticles. Optimization studies on the efficiency and specificity of the PEXT reaction were performed. The method was evaluated by genotyping 46 DNA samples of known genotype and 34 blind samples. The results were fully concordant with those obtained by reference methods. The method is simple, rapid, cost-effective and it does not require specialized instrumentation or highly qualified personnel. PMID:26580967

  20. Multi-allele genotyping platform for the simultaneous detection of mutations in the Wilson disease related ATP7B gene.

    PubMed

    Amvrosiadou, Maria; Petropoulou, Margarita; Poulou, Myrto; Tzetis, Maria; Kanavakis, Emmanuel; Christopoulos, Theodore K; Ioannou, Penelope C

    2015-12-01

    Wilson's disease is an inherited disorder of copper transport in the hepatocytes with a wide range of genotype and phenotype characteristics. Mutations in the ATP7B gene are responsible for the disease. Approximately, over 500 mutations in the ATP7B gene have been described to date. We report a method for the simultaneous detection of the ten most common ATP7B gene mutations in Greek patients. The method comprises 3 simple steps: (i) multiplex PCR amplification of fragments in the ATP7B gene flanking the mutations (ii) multiplex primer extension reaction of the unpurified amplification products using allele-specific primers and (iii) visual detection of the primer extension reaction products within minutes by means of dry-reagent multi-allele dipstick assay using anti-biotin conjugated gold nanoparticles. Optimization studies on the efficiency and specificity of the PEXT reaction were performed. The method was evaluated by genotyping 46 DNA samples of known genotype and 34 blind samples. The results were fully concordant with those obtained by reference methods. The method is simple, rapid, cost-effective and it does not require specialized instrumentation or highly qualified personnel.

  1. Functional assessment of the carboxy-terminus of the Wilson disease copper-transporting ATPase, ATP7B.

    PubMed

    Hsi, Gloria; Cullen, Lara M; Moira Glerum, D; Cox, Diane W

    2004-03-01

    The carboxy-terminus of ATP7B, the protein defective in the copper-transport disorder Wilson disease, was investigated with respect to its role in copper delivery to the ferroxidase ceruloplasmin. We use yeast as a model system to assess the functional capabilities of ATP7B variants. The yeast ferroxidase, Fet3p, acquires copper from Ccc2p and cannot function if Ccc2p is impaired; expression of wild-type ATP7B in ccc2 yeast complements the iron-deficient phenotype. Our results demonstrate that the C-terminus of ATP7B is necessary for protein stability, as removal of the nonmembranous terminus leads to reduced protein levels and cessation of growth in iron-limited medium. Growth is partially restored when an additional three amino acids are present and is near wild-type levels when only one-third of the C-terminus is present. Measurement of ferroxidase activity is a more sensitive indicator of copper transport function and allowed identification of impaired variants not detected with the growth assay. PMID:14962673

  2. Niemann-Pick C1 protein transports copper to the secretory compartment from late endosomes where ATP7B resides.

    PubMed

    Yanagimoto, Chikatoshi; Harada, Masaru; Kumemura, Hiroto; Koga, Hironori; Kawaguchi, Takumi; Terada, Kunihiko; Hanada, Shinichiro; Taniguchi, Eitaro; Koizumi, Yukio; Koyota, Souichi; Ninomiya, Haruaki; Ueno, Takato; Sugiyama, Toshihiro; Sata, Michio

    2009-01-15

    Wilson disease is a genetic disorder characterized by the accumulation of copper in the body by defective biliary copper excretion. Wilson disease gene product (ATP7B) functions in copper incorporation to ceruloplasmin (Cp) and biliary copper excretion. However, copper metabolism in hepatocytes has been still unclear. Niemann-Pick disease type C (NPC) is a lipid storage disorder and the most commonly mutated gene is NPC1 and its gene product NPC1 is a late endosome protein and regulates intracellular vesicle traffic. In the present study, we induced NPC phenotype and examined the localization of ATP7B and secretion of holo-Cp, a copper-binding mature form of Cp. The vesicle traffic was modulated using U18666A, which induces NPC phenotype, and knock down of NPC1 by RNA interference. ATP7B colocalized with the late endosome markers, but not with the trans-Golgi network markers. U18666A and NPC1 knock down decreased holo-Cp secretion to culture medium, but did not affect the secretion of other secretory proteins. Copper accumulated in the cells after the treatment with U18666A. These findings suggest that ATP7B localizes in the late endosomes and that copper in the late endosomes is transported to the secretory compartment via NPC1-dependent pathway and incorporated into apo-Cp to form holo-Cp.

  3. Fusion of the ZC3H7B and BCOR genes in endometrial stromal sarcomas carrying an X;22-translocation.

    PubMed

    Panagopoulos, Ioannis; Thorsen, Jim; Gorunova, Ludmila; Haugom, Lisbeth; Bjerkehagen, Bodil; Davidson, Ben; Heim, Sverre; Micci, Francesca

    2013-07-01

    Endometrial stromal sarcomas (ESS) are genetically heterogeneous uterine tumors in which a JAZF1-SUZ12 chimeric gene resulting from the chromosomal translocation t(7;17)(p15;q21) as well as PHF1 rearrangements (in chromosomal band 6p21) with formation of JAZF1-PHF1, EPC1-PHF1, and MEAF6-PHF1 chimeras have been described. Here, we investigated two ESS characterized cytogenetically by the presence of a der(22)t(X;22)(p11;q13). Whole transcriptome sequencing one of the tumors identified a ZC3H7-BCOR chimeric transcript. Reverse transciptase-PCR with the ZC3H7B forward and BCOR reverse primer combinations confirmed the presence of a ZC3H7-BCOR chimeric transcript in both ESS carrying a der(22)t(X;22) but not in a control ESS with t(1;6) and the MEAF6-PHF1 fusion. Sequencing of the amplified cDNA fragments showed that in both cases ESS exon 10 of ZC3H7B (from 22q13; accession number NM_017590 version 4) was fused to exon 8 of BCOR (from Xp11; accession number NM_001123385 version 1). Reciprocal multiple BCOR-ZC3H7B cDNA fragments were amplified in only one case suggesting that ZC3H7B-BCOR, on the der(22)t(X;22), is the pathogenetically important fusion gene. The putative ZC3H7B-BCOR protein would contain the tetratricopeptide repeats and LD motif from ZC3H7B and the AF9 binding site (1093-1233aa), the 3 ankyrin repeats (1410-1509 aa), and the NSPC1 binding site of BCOR. Although the presence of these motifs suggests various functions of the chimeric protein, it is possible that its most important role may be in epigenetic regulation. Whether or not the (patho)genetic subsets JAZF1-SUZ12, PHF1 rearrangements, and ZC3H7B-BCOR correspond to any phenotypic, let alone clinically important, differences in ESS remain unknown. PMID:23580382

  4. Aldehyde dehydrogenase 7A1 (ALDH7A1) attenuates reactive aldehyde and oxidative stress induced cytotoxicity

    PubMed Central

    Brocker, Chad; Cantore, Miriam; Failli, Paola; Vasiliou, Vasilis

    2012-01-01

    Mammalian aldehyde dehydrogenase 7A1 (ALDH7A1) is homologous to plant ALDH7B1 which protects against various forms of stress such as increased salinity, dehydration and treatment with oxidants or pesticides. Deleterious mutations in human ALDH7A1 are responsible for pyridoxine-dependent and folinic acid-responsive seizures. In previous studies, we have shown that human ALDH7A1 protects against hyperosmotic stress presumably through the generation of betaine, an important cellular osmolyte, formed from betaine aldehyde. Hyperosmotic stress is coupled to an increase in oxidative stress and lipid peroxidation (LPO). In this study, cell viability assays revealed that stable expression of mitochondrial ALDH7A1 in Chinese hamster ovary (CHO) cells provides significant protection against treatment with the LPO-derived aldehydes hexanal and 4-hydroxy-2-nonenal (4HNE) implicating a protective function for the enzyme during oxidative stress. A significant increase in cell survival was also observed in CHO cells expressing either mitochondrial or cytosolic ALDH7A1 treated with increasing concentrations of hydrogen peroxide (H2O2) or 4HNE, providing further evidence for anti-oxidant activity. In vitro enzyme activity assays indicate that human ALDH7A1 is sensitive to oxidation and that efficiency can be at least partially restored by incubating recombinant protein with the thiol reducing agent β-mercaptoethanol (BME). We also show that after reactivation with BME, recombinant ALDH7A1 is capable of metabolizing the reactive aldehyde 4HNE. In conclusion, ALDH7A1 mechanistically appears to provide cells protection through multiple pathways including the removal of toxic LPO-derived aldehydes in addition to osmolyte generation. PMID:21338592

  5. High CO2 Leads to Na,K-ATPase Endocytosis via c-Jun Amino-Terminal Kinase-Induced LMO7b Phosphorylation.

    PubMed

    Dada, Laura A; Trejo Bittar, Humberto E; Welch, Lynn C; Vagin, Olga; Deiss-Yehiely, Nimrod; Kelly, Aileen M; Baker, Mairead R; Capri, Joseph; Cohn, Whitaker; Whitelegge, Julian P; Vadász, István; Gruenbaum, Yosef; Sznajder, Jacob I

    2015-12-01

    The c-Jun amino-terminal kinase (JNK) plays a role in inflammation, proliferation, apoptosis, and cell adhesion and cell migration by phosphorylating paxillin and β-catenin. JNK phosphorylation downstream of AMP-activated protein kinase (AMPK) activation is required for high CO2 (hypercapnia)-induced Na,K-ATPase endocytosis in alveolar epithelial cells. Here, we provide evidence that during hypercapnia, JNK promotes the phosphorylation of LMO7b, a scaffolding protein, in vitro and in intact cells. LMO7b phosphorylation was blocked by exposing the cells to the JNK inhibitor SP600125 and by infecting cells with dominant-negative JNK or AMPK adenovirus. The knockdown of the endogenous LMO7b or overexpression of mutated LMO7b with alanine substitutions of five potential JNK phosphorylation sites (LMO7b-5SA) or only Ser-1295 rescued both LMO7b phosphorylation and the hypercapnia-induced Na,K-ATPase endocytosis. Moreover, high CO2 promoted the colocalization and interaction of LMO7b and the Na,K-ATPase α1 subunit at the plasma membrane, which were prevented by SP600125 or by transfecting cells with LMO7b-5SA. Collectively, our data suggest that hypercapnia leads to JNK-induced LMO7b phosphorylation at Ser-1295, which facilitates the interaction of LMO7b with Na,K-ATPase at the plasma membrane promoting the endocytosis of Na,K-ATPase in alveolar epithelial cells.

  6. High CO2 Leads to Na,K-ATPase Endocytosis via c-Jun Amino-Terminal Kinase-Induced LMO7b Phosphorylation

    PubMed Central

    Trejo Bittar, Humberto E.; Welch, Lynn C.; Vagin, Olga; Deiss-Yehiely, Nimrod; Kelly, Aileen M.; Baker, Mairead R.; Capri, Joseph; Cohn, Whitaker; Whitelegge, Julian P.; Vadász, István; Gruenbaum, Yosef; Sznajder, Jacob I.

    2015-01-01

    The c-Jun amino-terminal kinase (JNK) plays a role in inflammation, proliferation, apoptosis, and cell adhesion and cell migration by phosphorylating paxillin and β-catenin. JNK phosphorylation downstream of AMP-activated protein kinase (AMPK) activation is required for high CO2 (hypercapnia)-induced Na,K-ATPase endocytosis in alveolar epithelial cells. Here, we provide evidence that during hypercapnia, JNK promotes the phosphorylation of LMO7b, a scaffolding protein, in vitro and in intact cells. LMO7b phosphorylation was blocked by exposing the cells to the JNK inhibitor SP600125 and by infecting cells with dominant-negative JNK or AMPK adenovirus. The knockdown of the endogenous LMO7b or overexpression of mutated LMO7b with alanine substitutions of five potential JNK phosphorylation sites (LMO7b-5SA) or only Ser-1295 rescued both LMO7b phosphorylation and the hypercapnia-induced Na,K-ATPase endocytosis. Moreover, high CO2 promoted the colocalization and interaction of LMO7b and the Na,K-ATPase α1 subunit at the plasma membrane, which were prevented by SP600125 or by transfecting cells with LMO7b-5SA. Collectively, our data suggest that hypercapnia leads to JNK-induced LMO7b phosphorylation at Ser-1295, which facilitates the interaction of LMO7b with Na,K-ATPase at the plasma membrane promoting the endocytosis of Na,K-ATPase in alveolar epithelial cells. PMID:26370512

  7. The Wilson disease protein ATP7B resides in the late endosomes with Rab7 and the Niemann-Pick C1 protein.

    PubMed

    Harada, Masaru; Kawaguchi, Takumi; Kumemura, Hiroto; Terada, Kunihiko; Ninomiya, Haruaki; Taniguchi, Eitaro; Hanada, Shinichiro; Baba, Shinji; Maeyama, Michiko; Koga, Hironori; Ueno, Takato; Furuta, Koh; Suganuma, Tatsuo; Sugiyama, Toshihiro; Sata, Michio

    2005-02-01

    Wilson disease is a genetic disorder characterized by the accumulation of copper in the body due to a defect of biliary copper excretion. Although the Wilson disease gene has been cloned, the cellular localization of the gene product (ATP7B) has not been fully clarified. Therefore, the precise physiological action of ATP7B is still unknown. We examined the distribution of ATP7B using an anti-ATP7B antibody, green fluorescent protein (GFP)-ATP7B (GFP-ATP7B) and ATP7B-DsRed in various cultured cells. Intracellular organelles were visualized by fluorescence microscopy. The distribution of ATP7B was compared with that of Rab7 and Niemann-Pick C1 (NPC1), proteins that localize in the late endosomes. U18666A, which induces the NPC phenotype, was used to modulate the intracellular vesicle traffic. GFP-ATP7B colocalized with various late endosome markers including Rab7 and NPC1 but not with Golgi or lysosome markers. U18666A induced the formation of late endosome-lysosome hybrid organelles, with GFP-ATP7B localized with NPC1 in these structures. We have confirmed that ATP7B is a late endosome-associated membrane protein. ATP7B appears to translocate copper from the cytosol to the late endosomal lumen, thus participating in biliary copper excretion via lysosomes. Thus, defective copper ATPase activity of ATP7B in the late endosomes appears to be the main defect of Wilson disease.

  8. 7C2, the new neutron diffractometer for liquids and disordered materials at LLB

    NASA Astrophysics Data System (ADS)

    Cuello, G. J.; Darpentigny, J.; Hennet, L.; Cormier, L.; Dupont, J.; Homatter, B.; Beuneu, B.

    2016-09-01

    The disordered-materials diffractometer 7C2 at the Laboratoire Leon Brillouin (LLB), Saclay, France has been upgraded through the replacement of its detector. The old one, a banana like BF3 detector, has been replaced by a new ensemble of 3He tubes arranged on a cylindrical surface concentric with the sample position, covering an angular range of 133°, with an angular step of 0.52°. At the shortest wavelength, 0.57 Å, this represents a maximum momentum transfer of 20 Å-1. The gas pressure in the 3He tubes is 30 bars, which increases the efficiency to 80% for 0.72 Å neutrons, compared to 17% for the previous detector. A larger solid angle of detection and an improved efficiency have increased the total counting rate by a factor of 22, thereby reducing random error in the diffraction measurements. The banana-like multidetector gives to 7C2 the possibility of registering a complete diffractogram over the whole angular range in one shot, which is a comparative advantage with respect to other hot-neutrons two-axis diffractometers.

  9. The Radio luminosity Function of Radio-Loud Quasars from the 7C Redshift Survey

    NASA Technical Reports Server (NTRS)

    Willott, Chris J.; Rawlings, Steve; Blundell, Katherine M.; Lacy, Mark

    1998-01-01

    We present a complete sample of 24 radio-loud quasars (RLQs) from the new 7C Redshift Survey. Every quasar with a low-frequency (151 MHz) radio flux-density S(sub 151) > 0.5 Jy in two regions of the sky covering 0.013 sr is included; 23 of these have sufficient extended flux to meet the selection criteria, 18 of these have steep radio spectra (hereafter denoted as SSQs). The key advantage of this sample over most samples of RLQs is the lack of an optical magnitude limit. By combining the 7C and 3CRR samples, we have investigated the properties of RLQs as a function of redshift z and radio luminosity L(sub 151). We derive the radio luminosity function (RLF) of RLQs and find that the data are well fitted by a single power-law with slope alpha(sub 1) = 1.9 +/- 0.1 (for H(sub 0) = 50 km/s.Mpc, OMEGA(sub M) = 1, OMEGA(sub DELTA) = 0). We find that there must be a break in the RLQ RLF at log(sub 10)(L(sub 151)/W Hz.sr) approximately < or = 27, in order for the models to be consistent with the 7C and 6C source counts. The z-dependence of the RLF follows a one-tailed gaussian which peaks at z = 1.7 +/- 0.2. We find no evidence for a decline in the co-moving space density of RLQs at higher redshifts. A positive correlation between the radio and optical luminosities of SSQs is observed, confirming a result of Serjeant. We are able to rule out this correlation being due to selection effects or biases in our combined sample. The radio-optical correlation and best-fit model RLF enable us to estimate the distribution of optical magnitudes of quasars in samples selected at low radio frequencies, We con- clude that for samples with S(sub 151) approximately < or = 1 Jy one must use optical data significantly deeper than the POSS-I limit (R approximately equal 20), in order to avoid severe incompleteness.

  10. Recombinant Trichoderma harzianum endoglucanase I (Cel7B) is a highly acidic and promiscuous carbohydrate-active enzyme.

    PubMed

    Pellegrini, Vanessa O A; Serpa, Viviane Isabel; Godoy, Andre S; Camilo, Cesar M; Bernardes, Amanda; Rezende, Camila A; Junior, Nei Pereira; Franco Cairo, João Paulo L; Squina, Fabio M; Polikarpov, Igor

    2015-11-01

    Trichoderma filamentous fungi have been investigated due to their ability to secrete cellulases which find various biotechnological applications such as biomass hydrolysis and cellulosic ethanol production. Previous studies demonstrated that Trichoderma harzianum IOC-3844 has a high degree of cellulolytic activity and potential for biomass hydrolysis. However, enzymatic, biochemical, and structural studies of cellulases from T. harzianum are scarce. This work reports biochemical characterization of the recombinant endoglucanase I from T. harzianum, ThCel7B, and its catalytic core domain. The constructs display optimum activity at 55 °C and a surprisingly acidic pH optimum of 3.0. The full-length enzyme is able to hydrolyze a variety of substrates, with high specific activity: 75 U/mg for β-glucan, 46 U/mg toward xyloglucan, 39 U/mg for lichenan, 26 U/mg for carboxymethyl cellulose, 18 U/mg for 4-nitrophenyl β-D-cellobioside, 16 U/mg for rye arabinoxylan, and 12 U/mg toward xylan. The enzyme also hydrolyzed filter paper, phosphoric acid swollen cellulose, Sigmacell 20, Avicel PH-101, and cellulose, albeit with lower efficiency. The ThCel7B catalytic domain displays similar substrate diversity. Fluorescence-based thermal shift assays showed that thermal stability is highest at pH 5.0. We determined kinetic parameters and analyzed a pattern of oligosaccharide substrates hydrolysis, revealing cellobiose as a final product of C6 degradation. Finally, we visualized effects of ThCel7B on oat spelt using scanning electron microscopy, demonstrating the morphological changes of the substrate during the hydrolysis. The acidic behavior of ThCel7B and its considerable thermostability hold a promise of its industrial applications and other biotechnological uses under extremely acidic conditions. PMID:26156238

  11. Distinct phenotype of a Wilson disease mutation reveals a novel trafficking determinant in the copper transporter ATP7B

    PubMed Central

    Braiterman, Lelita T.; Murthy, Amrutha; Jayakanthan, Samuel; Nyasae, Lydia; Tzeng, Eric; Gromadzka, Grazyna; Woolf, Thomas B.; Lutsenko, Svetlana; Hubbard, Ann L.

    2014-01-01

    Wilson disease (WD) is a monogenic autosomal-recessive disorder of copper accumulation that leads to liver failure and/or neurological deficits. WD is caused by mutations in ATP7B, a transporter that loads Cu(I) onto newly synthesized cupro-enzymes in the trans-Golgi network (TGN) and exports excess copper out of cells by trafficking from the TGN to the plasma membrane. To date, most WD mutations have been shown to disrupt ATP7B activity and/or stability. Using a multidisciplinary approach, including clinical analysis of patients, cell-based assays, and computational studies, we characterized a patient mutation, ATP7BS653Y, which is stable, does not disrupt Cu(I) transport, yet renders the protein unable to exit the TGN. Bulky or charged substitutions at position 653 mimic the phenotype of the patient mutation. Molecular modeling and dynamic simulation suggest that the S653Y mutation induces local distortions within the transmembrane (TM) domain 1 and alter TM1 interaction with TM2. S653Y abolishes the trafficking-stimulating effects of a secondary mutation in the N-terminal apical targeting domain. This result indicates a role for TM1/TM2 in regulating conformations of cytosolic domains involved in ATP7B trafficking. Taken together, our experiments revealed an unexpected role for TM1/TM2 in copper-regulated trafficking of ATP7B and defined a unique class of WD mutants that are transport-competent but trafficking-defective. Understanding the precise consequences of WD-causing mutations will facilitate the development of advanced mutation-specific therapies. PMID:24706876

  12. The Effect of Zinc and D-Penicillamine in a Stable Human Hepatoma ATP7B Knockout Cell Line

    PubMed Central

    Chandhok, Gursimran; Schmitt, Nadine; Sauer, Vanessa; Aggarwal, Annu; Bhatt, Mohit; Schmidt, Hartmut H. J.

    2014-01-01

    Mutations in the copper (Cu) transporter gene ATP7B, the primary cause of Wilson disease (WD), result in high liver Cu and death of hepatocytes. Cu chelators and zinc salts are the two most important drugs used in the treatment of WD patients; however, the molecular mechanisms of the drugs with regard to ATP7B expression have not been determined. A targeted knockout of ATP7B (KO) was established in the most widely used human hepatoma cell line, HepG2 for molecular studies of the pathogenesis and treatment of the disease. KO cells showed similar growth, Cu uptake, release, and gene expression as compared to parental cells. However, in the presence of Cu, morphological changes, oxidative stress, apoptosis, and loss of viability were observed. Induction of metallothionein (MT1X) after Cu exposure was significantly reduced in KO cells. Following zinc treatment, MT1X expression was strongly induced and a high percentage of KO cells could be rescued from Cu induced toxicity. D-penicillamine treatment had a minor effect on the viability of KO cells whereas the parental cell line showed a pronounced improvement. Combined treatment displayed a highly synergistic effect in KO cells. The data suggest that zinc has a previously unrecognized effect on the viability of hepatocytes that lack ATP7B due to a high induction of MT1X expression that compensates low gene expression after Cu exposure. A combination therapy that simultaneously targets at MT1X induction and Cu chelation improves the overall survival of hepatocytes for most efficient therapy of patients having WD. PMID:24892424

  13. Recombinant Trichoderma harzianum endoglucanase I (Cel7B) is a highly acidic and promiscuous carbohydrate-active enzyme.

    PubMed

    Pellegrini, Vanessa O A; Serpa, Viviane Isabel; Godoy, Andre S; Camilo, Cesar M; Bernardes, Amanda; Rezende, Camila A; Junior, Nei Pereira; Franco Cairo, João Paulo L; Squina, Fabio M; Polikarpov, Igor

    2015-11-01

    Trichoderma filamentous fungi have been investigated due to their ability to secrete cellulases which find various biotechnological applications such as biomass hydrolysis and cellulosic ethanol production. Previous studies demonstrated that Trichoderma harzianum IOC-3844 has a high degree of cellulolytic activity and potential for biomass hydrolysis. However, enzymatic, biochemical, and structural studies of cellulases from T. harzianum are scarce. This work reports biochemical characterization of the recombinant endoglucanase I from T. harzianum, ThCel7B, and its catalytic core domain. The constructs display optimum activity at 55 °C and a surprisingly acidic pH optimum of 3.0. The full-length enzyme is able to hydrolyze a variety of substrates, with high specific activity: 75 U/mg for β-glucan, 46 U/mg toward xyloglucan, 39 U/mg for lichenan, 26 U/mg for carboxymethyl cellulose, 18 U/mg for 4-nitrophenyl β-D-cellobioside, 16 U/mg for rye arabinoxylan, and 12 U/mg toward xylan. The enzyme also hydrolyzed filter paper, phosphoric acid swollen cellulose, Sigmacell 20, Avicel PH-101, and cellulose, albeit with lower efficiency. The ThCel7B catalytic domain displays similar substrate diversity. Fluorescence-based thermal shift assays showed that thermal stability is highest at pH 5.0. We determined kinetic parameters and analyzed a pattern of oligosaccharide substrates hydrolysis, revealing cellobiose as a final product of C6 degradation. Finally, we visualized effects of ThCel7B on oat spelt using scanning electron microscopy, demonstrating the morphological changes of the substrate during the hydrolysis. The acidic behavior of ThCel7B and its considerable thermostability hold a promise of its industrial applications and other biotechnological uses under extremely acidic conditions.

  14. An electrochemical biosensor for double-stranded Wnt7B gene detection based on enzymatic isothermal amplification.

    PubMed

    Li, Junlong; Chen, Zhongping; Xiang, Yu; Zhou, Lili; Wang, Ting; Zhang, Zhang; Sun, Kexin; Yin, Dan; Li, Yi; Xie, Guoming

    2016-12-15

    Wnt7B gene plays an important role in the development and progression of breast cancer, gastric cancer, esophageal cancer and pancreatic cancer. While, the natural state of DNA is double stranded, which makes it difficult to be directly detected. Here, we develop an electrochemical biosensor method for Wnt7B gene detection without the need to denature the target. This method firstly used nicking enzyme for exploiting in the double-stranded DNA (dsDNA). Then, long single-stranded DNA (ssDNA) was generated from the cutting site through polymerase extension reaction. Whereafter, the long ssDNA triggered a hairpin self-assembly recycling reaction, which gave rise to another isothermal amplification reaction. Last, short ssDNA was formed after the this amplification process, which could hybridize with the capture probe immobilized on Au electrode and result in signal variation. This method showed excellent analytical performance for dsDNA, of which the linear range was 2fM to 500pM and the detection limit was 1.6fM (S/N=3). It also showed an good results when applied to the real sample of Wnt7B gene detection. PMID:27326913

  15. Spin-orbit alignment for KELT-7b and HAT-P-56b via Doppler tomography with TRES

    NASA Astrophysics Data System (ADS)

    Zhou, George; Latham, David W.; Bieryla, Allyson; Beatty, Thomas G.; Buchhave, Lars A.; Esquerdo, Gilbert A.; Berlind, Perry; Calkins, Michael L.

    2016-08-01

    We present Doppler tomographic analyses for the spectroscopic transits of KELT-7b and HAT-P-56b, two hot-Jupiters orbiting rapidly rotating F-dwarf host stars. These include analyses of archival Tillinghast Reflector Echelle Spectrograph (TRES) observations for KELT-7b, and a new TRES transit observation of HAT-P-56b. We report spin-orbit aligned geometries for KELT-7b (2.7° ± 0.6°) and HAT-P-56b (8° ± 2°). The host stars KELT-7 and HAT-P-56 are among some of the most rapidly rotating planet-hosting stars known. We examine the tidal re-alignment model for the evolution of the spin-orbit angle in the context of the spin rates of these stars. We find no evidence that the rotation rates of KELT-7 and HAT-P-56 have been modified by star-planet tidal interactions, suggesting that the spin-orbit angle of systems around these hot stars may represent their primordial configuration. In fact, KELT-7 and HAT-P-56 are two of three systems in supersynchronous, spin-orbit aligned states, where the rotation periods of the host stars are faster than the orbital periods of the planets.

  16. Intronic rs2147363 variant in ATP7B transcription factor-binding site associated with Alzheimer's disease.

    PubMed

    Bucossi, Serena; Polimanti, Renato; Ventriglia, Mariacarla; Mariani, Stefania; Siotto, Mariacristina; Ursini, Francesca; Trotta, Laura; Scrascia, Federica; Callea, Antonio; Vernieri, Fabrizio; Squitti, Rosanna

    2013-01-01

    Copper homeostasis abnormalities have been shown to be associated with Alzheimer's disease (AD), possibly by accelerating amyloid-β toxicity and plaque formation. The ATP7B gene plays a key role in controlling body copper balance. Our previous studies showed an association between ATP7B variants and AD risk. Among these variants, an intronic single nucleotide polymorphism, rs2147363, was associated with AD risk. In order to understand this intronic association, we screened a population of 286 AD patients and 283 healthy controls, and verified the presence of other functional coding variants in linkage disequilibrium (LD). Then we searched for a regulatory function region close to rs2147363. An LD analysis revealed the presence of an LD between rs2147363 and a Wilson's disease-causing variant, rs7334118. However, this mutation did not explain the observed genetic association. Conversely, in silico analyses of rs2147363 functionality highlighted that this variant is located in a binding site of a transcription factor, and is, consequently, associated with regulatory function. These data suggest that the genetic variation in cis-regulatory elements located in non-coding regions can have a role in determining ATP7B functionality and account for some of the AD missing hereditability.

  17. Solvothermal synthesis of Fe7C3 and Fe3C nanostructures with phase and morphology control

    NASA Astrophysics Data System (ADS)

    Williams, Brent; Clifford, Dustin; El-Gendy, Ahmed A.; Carpenter, Everett E.

    2016-07-01

    A phase transition, from orthorhombic Fe3C to hexagonal Fe7C3, was observed using a wet synthesis mediated by hexadecyltrimethylammonium chloride (CTAC). In this study, CTAC has been shown to control carbide phase, morphology, and size of the iron carbide nanostructures. Fe7C3 hexagonal prisms were formed with an average diameter of 960 nm, the thickness of 150 nm, and Fe3C nanostructures with an approximate size of 50 nm. Magnetic studies show ferromagnetic behavior with Ms of 126 emu/g, and Hc of 170 Oe with respect to Fe7C3 and 95 emu/g and 590 Oe with respect to Fe3C. The thermal studies using high temperature x-ray diffraction show stability of Fe7C3 up to 500 °C. Upon slow cooling, the Fe7C3 phase is recovered with an intermediate oxide phase occurring around 300 °C. This study has demonstrated a simple route in synthesizing iron carbides for an in depth magnetic study and crystal phase transition study of Fe7C3 at elevated temperatures.

  18. Effect of Ti additive on (Cr, Fe)7C3 carbide in arc surfacing layer and its refined mechanism

    NASA Astrophysics Data System (ADS)

    Zhou, Yefei; Yang, Yulin; Yang, Jian; Hao, Feifei; Li, Da; Ren, Xuejun; Yang, Qingxiang

    2012-06-01

    Arc surfacing layer of hypoeutectic high chromium cast iron (HCCI) expects refiner carbides in the microstructure to improve its mechanical properties. In this paper, Ti additive as a strong carbide forming element was added in the hypoeutectic HCCI arc surfacing layer. Microstructure of titaniferous hypoeutectic HCCI was studied by optical microscopy, X-ray diffraction and field emission scanning electronic microscopy with energy dispersive spectrometer. Furthermore, the M(M = Cr, Fe)7C3 carbide refinement mechanism was explained by the phase diagram calculation and lattice misfit theory. The results show that, the M7C3 carbide in arc surfacing microstructure of hypoeutectic HCCI has been refined with 2 wt.% Ti additive, and TiC carbide can be observed in/around the M7C3 carbide. With Ti addictive increasing, the micro-hardness along the depth in profile section of layer becomes more uniform, and the wear resistance has been improved. According to the phase diagram calculation, MC carbide precipitates prior to M7C3 carbide in Fe-Cr-C-Ti alloy. In addition, the lattice misfit between (1 1 0)TiC and (0C is 9.257%, which indicates that the TiC as heterogeneous nuclei of the M7C3 is medium effective. Therefore, the M7C3 carbide can be refined.

  19. Fairchild 7A compressor model

    SciTech Connect

    Foster, R.E.

    1986-09-23

    The Fairchild 7A centrifugal compressor used in the X-326 isotopic ''top'' cascade at the Portsmouth Gaseous Diffusion Plant has been modeled using a proprietary computer code called COMPAL by Concepts E.T.I., Inc. of Norwich, VT. The 7A compressor is described and the results of the modeling calculations are presented. Performance characteristics curves (PR vs. flow) are included for various mixtures of UF/sub 6/ and N/sub 2/ gases.

  20. The P7C3 class of neuroprotective compounds exerts antidepressant efficacy in mice by increasing hippocampal neurogenesis.

    PubMed

    Walker, A K; Rivera, P D; Wang, Q; Chuang, J-C; Tran, S; Osborne-Lawrence, S; Estill, S J; Starwalt, R; Huntington, P; Morlock, L; Naidoo, J; Williams, N S; Ready, J M; Eisch, A J; Pieper, A A; Zigman, J M

    2015-04-01

    Augmenting hippocampal neurogenesis represents a potential new strategy for treating depression. Here we test this possibility by comparing hippocampal neurogenesis in depression-prone ghrelin receptor (Ghsr)-null mice to that in wild-type littermates and by determining the antidepressant efficacy of the P7C3 class of neuroprotective compounds. Exposure of Ghsr-null mice to chronic social defeat stress (CSDS) elicits more severe depressive-like behavior than in CSDS-exposed wild-type littermates, and exposure of Ghsr-null mice to 60% caloric restriction fails to elicit antidepressant-like behavior. CSDS resulted in more severely reduced cell proliferation and survival in the ventral dentate gyrus (DG) subgranular zone of Ghsr-null mice than in that of wild-type littermates. Also, caloric restriction increased apoptosis of DG subgranular zone cells in Ghsr-null mice, although it had the opposite effect in wild-type littermates. Systemic treatment with P7C3 during CSDS increased survival of proliferating DG cells, which ultimately developed into mature (NeuN+) neurons. Notably, P7C3 exerted a potent antidepressant-like effect in Ghsr-null mice exposed to either CSDS or caloric restriction, while the more highly active analog P7C3-A20 also exerted an antidepressant-like effect in wild-type littermates. Focal ablation of hippocampal stem cells with radiation eliminated this antidepressant effect, further attributing the P7C3 class antidepressant effect to its neuroprotective properties and resultant augmentation of hippocampal neurogenesis. Finally, P7C3-A20 demonstrated greater proneurogenic efficacy than a wide spectrum of currently marketed antidepressant drugs. Taken together, our data confirm the role of aberrant hippocampal neurogenesis in the etiology of depression and suggest that the neuroprotective P7C3-compounds represent a novel strategy for treating patients with this disease.

  1. The P7C3 class of neuroprotective compounds exerts antidepressant efficacy in mice by increasing hippocampal neurogenesis

    PubMed Central

    Walker, AK; Rivera, PD; Wang, Q; Chuang, J-C; Tran, S; Osborne-Lawrence, S; Estill, SJ; Starwalt, R; Huntington, P; Morlock, L; Naidoo, J; Williams, NS; Ready, JM; Eisch, AJ; Pieper, AA; Zigman, JM

    2014-01-01

    Augmenting hippocampal neurogenesis represents a potential new strategy for treating depression. Here we test this possibility by comparing hippocampal neurogenesis in depression-prone ghrelin receptor (Ghsr)-null mice to that in wild-type littermates and by determining the antidepressant efficacy of the P7C3 class of neuroprotective compounds. Exposure of Ghsr-null mice to chronic social defeat stress (CSDS) elicits more severe depressive-like behavior than in CSDS-exposed wild-type littermates, and exposure of Ghsr-null mice to 60% caloric restriction fails to elicit antidepressant-like behavior. CSDS resulted in more severely reduced cell proliferation and survival in the ventral dentate gyrus (DG) subgranular zone of Ghsr-null mice than in that of wild-type littermates. Also, caloric restriction increased apoptosis of DG subgranular zone cells in Ghsr-null mice, although it had the opposite effect in wild-type littermates. Systemic treatment with P7C3 during CSDS increased survival of proliferating DG cells, which ultimately developed into mature (NeuN+) neurons. Notably, P7C3 exerted a potent antidepressant-like effect in Ghsr-null mice exposed to either CSDS or caloric restriction, while the more highly active analog P7C3-A20 also exerted an antidepressant-like effect in wild-type littermates. Focal ablation of hippocampal stem cells with radiation eliminated this antidepressant effect, further attributing the P7C3 class antidepressant effect to its neuroprotective properties and resultant augmentation of hippocampal neurogenesis. Finally, P7C3-A20 demonstrated greater proneurogenic efficacy than a wide spectrum of currently marketed antidepressant drugs. Taken together, our data confirm the role of aberrant hippocampal neurogenesis in the etiology of depression and suggest that the neuroprotective P7C3-compounds represent a novel strategy for treating patients with this disease. PMID:24751964

  2. First-principles calculations of properties of orthorhombic iron carbide Fe7C3 at the Earth's core conditions

    NASA Astrophysics Data System (ADS)

    Raza, Zamaan; Shulumba, Nina; Caffrey, Nuala M.; Dubrovinsky, Leonid; Abrikosov, Igor A.

    2015-06-01

    A recently discovered phase of orthorhombic iron carbide o-Fe7C3 [Prescher et al., Nat. Geosci. 8, 220 (2015), 10.1038/ngeo2370] is assessed as a potentially important phase for interpretation of the properties of the Earth's core. In this paper, we carry out first-principles calculations on o-Fe7C3 , finding properties to be in broad agreement with recent experiments, including a high Poisson's ratio (0.38). Our enthalpy calculations suggest that o-Fe7C3 is more stable than Eckstrom-Adcock hexagonal iron carbide (h-Fe7C3 ) below approximately 100 GPa. However, at 150 GPa, the two phases are essentially degenerate in terms of Gibbs free energy, and further increasing the pressure towards Earth's core conditions stabilizes h-Fe7C3 with respect to the orthorhombic phase. Increasing the temperature tends to stabilize the hexagonal phase at 360 GPa, but this trend may change beyond the limit of the quasiharmonic approximation.

  3. P7C3 Attenuates the Scopolamine-Induced Memory Impairments in C57BL/6J Mice.

    PubMed

    Jiang, Bo; Song, Lu; Huang, Chao; Zhang, Wei

    2016-05-01

    Memory impairment is the most common symptom in patients with Alzheimer's disease. The purpose of this study is to evaluate the memory enhancing effects of P7C3, a recently identified compound with robust proneurogenic and neuroprotective effects, on the cognitive impairment induced by scopolamine, a muscarinic acetylcholine receptor antagonist. Different behavior tests including the Y-maze, Morris water maze, and passive avoidance tests were performed to measure cognitive functions. Scopolamine significantly decreased the spontaneous alternation and step-through latency of C57BL/6J mice in Y-maze test and passive avoidance test, whereas increased the time of mice spent to find the hidden platform in Morris water maze test. Importantly, intraperitoneal administration of P7C3 effectively reversed those Scopolamine-induced cognitive impairments in C57BL/6J mice. Furthermore, P7C3 treatment significantly enhanced the level of brain-derived neurotrophic factor (BDNF) signaling pathway in the cortex and hippocampus, and the usage of selective BDNF signaling inhibitor fully blocked the anti-amnesic effects of P7C3. Therefore, these findings suggest that P7C3 could improve the scopolamine-induced learning and memory impairment possibly through activation of BDNF signaling pathway, thereby exhibiting a cognition-enhancing potential.

  4. Could CoRoT-7b and Kepler-10b be remnants of evaporated gas or ice giants?

    PubMed

    Leitzinger, M; Odert, P; Kulikov, Yu N; Lammer, H; Wuchterl, G; Penz, T; Guarcello, M G; Micela, G; Khodachenko, M L; Weingrill, J; Hanslmeier, A; Biernat, H K; Schneider, J

    2011-10-01

    We present thermal mass loss calculations over evolutionary time scales for the investigation if the smallest transiting rocky exoplanets CoRoT-7b (∼1.68REarth) and Kepler-10b (∼1.416REarth) could be remnants of an initially more massive hydrogen-rich gas giant or a hot Neptune-class exoplanet. We apply a thermal mass loss formula which yields results that are comparable to hydrodynamic loss models. Our approach considers the effect of the Roche lobe, realistic heating efficiencies and a radius scaling law derived from observations of hot Jupiters. We study the influence of the mean planetary density on the thermal mass loss by placing hypothetical exoplanets with the characteristics of Jupiter, Saturn, Neptune, and Uranus to the orbital location of CoRoT-7b at 0.017 AU and Kepler-10b at 0.01684 AU and assuming that these planets orbit a K- or G-type host star. Our findings indicate that hydrogen-rich gas giants within the mass domain of Saturn or Jupiter cannot thermally lose such an amount of mass that CoRoT-7b and Kepler-10b would result in a rocky residue. Moreover, our calculations show that the present time mass of both rocky exoplanets can be neither a result of evaporation of a hydrogen envelope of a "Hot Neptune" nor a "Hot Uranus"-class object. Depending on the initial density and mass, these planets most likely were always rocky planets which could lose a thin hydrogen envelope, but not cores of thermally evaporated initially much more massive and larger objects. PMID:21969736

  5. Could CoRoT-7b and Kepler-10b be remnants of evaporated gas or ice giants?

    PubMed Central

    Leitzinger, M.; Odert, P.; Kulikov, Yu.N.; Lammer, H.; Wuchterl, G.; Penz, T.; Guarcello, M.G.; Micela, G.; Khodachenko, M.L.; Weingrill, J.; Hanslmeier, A.; Biernat, H.K.; Schneider, J.

    2011-01-01

    We present thermal mass loss calculations over evolutionary time scales for the investigation if the smallest transiting rocky exoplanets CoRoT-7b (∼1.68REarth) and Kepler-10b (∼1.416REarth) could be remnants of an initially more massive hydrogen-rich gas giant or a hot Neptune-class exoplanet. We apply a thermal mass loss formula which yields results that are comparable to hydrodynamic loss models. Our approach considers the effect of the Roche lobe, realistic heating efficiencies and a radius scaling law derived from observations of hot Jupiters. We study the influence of the mean planetary density on the thermal mass loss by placing hypothetical exoplanets with the characteristics of Jupiter, Saturn, Neptune, and Uranus to the orbital location of CoRoT-7b at 0.017 AU and Kepler-10b at 0.01684 AU and assuming that these planets orbit a K- or G-type host star. Our findings indicate that hydrogen-rich gas giants within the mass domain of Saturn or Jupiter cannot thermally lose such an amount of mass that CoRoT-7b and Kepler-10b would result in a rocky residue. Moreover, our calculations show that the present time mass of both rocky exoplanets can be neither a result of evaporation of a hydrogen envelope of a “Hot Neptune” nor a “Hot Uranus”-class object. Depending on the initial density and mass, these planets most likely were always rocky planets which could lose a thin hydrogen envelope, but not cores of thermally evaporated initially much more massive and larger objects. PMID:21969736

  6. Could CoRoT-7b and Kepler-10b be remnants of evaporated gas or ice giants?

    PubMed

    Leitzinger, M; Odert, P; Kulikov, Yu N; Lammer, H; Wuchterl, G; Penz, T; Guarcello, M G; Micela, G; Khodachenko, M L; Weingrill, J; Hanslmeier, A; Biernat, H K; Schneider, J

    2011-10-01

    We present thermal mass loss calculations over evolutionary time scales for the investigation if the smallest transiting rocky exoplanets CoRoT-7b (∼1.68REarth) and Kepler-10b (∼1.416REarth) could be remnants of an initially more massive hydrogen-rich gas giant or a hot Neptune-class exoplanet. We apply a thermal mass loss formula which yields results that are comparable to hydrodynamic loss models. Our approach considers the effect of the Roche lobe, realistic heating efficiencies and a radius scaling law derived from observations of hot Jupiters. We study the influence of the mean planetary density on the thermal mass loss by placing hypothetical exoplanets with the characteristics of Jupiter, Saturn, Neptune, and Uranus to the orbital location of CoRoT-7b at 0.017 AU and Kepler-10b at 0.01684 AU and assuming that these planets orbit a K- or G-type host star. Our findings indicate that hydrogen-rich gas giants within the mass domain of Saturn or Jupiter cannot thermally lose such an amount of mass that CoRoT-7b and Kepler-10b would result in a rocky residue. Moreover, our calculations show that the present time mass of both rocky exoplanets can be neither a result of evaporation of a hydrogen envelope of a "Hot Neptune" nor a "Hot Uranus"-class object. Depending on the initial density and mass, these planets most likely were always rocky planets which could lose a thin hydrogen envelope, but not cores of thermally evaporated initially much more massive and larger objects.

  7. STUDYING THE ATMOSPHERE OF THE EXOPLANET HAT-P-7b VIA SECONDARY ECLIPSE MEASUREMENTS WITH EPOXI, SPITZER, AND KEPLER

    SciTech Connect

    Christiansen, Jessie L.; Ballard, Sarah; Charbonneau, David; Holman, Matthew J.; Madhusudhan, Nikku; Seager, Sara; Wellnitz, Dennis D.; Deming, Drake; A'Hearn, Michael F.

    2010-02-10

    The highly irradiated transiting exoplanet, HAT-P-7b, currently provides one of the best opportunities for studying planetary emission in the optical and infrared wavelengths. We observe six near-consecutive secondary eclipses of HAT-P-7b at optical wavelengths with the EPOXI spacecraft. We place an upper limit on the relative eclipse depth of 0.055% (95% confidence). We also analyze Spitzer observations of the same target in the infrared, obtaining secondary eclipse depths of 0.098% +- 0.017%, 0.159% +- 0.022%, 0.245% +- 0.031%, and 0.225% +- 0.052% in the 3.6, 4.5, 5.8, and 8.0 {mu}m IRAC bands, respectively. We combine these measurements with the recently published Kepler secondary eclipse measurement and generate atmospheric models for the dayside of the planet that are consistent with both the optical and infrared measurements. The data are best fit by models with a temperature inversion, as expected from the high incident flux. The models predict a low optical albedo of {approx}<0.13, with subsolar abundances of Na, K, TiO, and VO. We also find that the best-fitting models predict that 10% of the absorbed stellar flux is redistributed to the nightside of the planet, which is qualitatively consistent with the inefficient day-night redistribution apparent in the Kepler phase curve. Models without thermal inversions fit the data only at the 1.25sigma level, and also require an overabundance of methane, which is not expected in the very hot atmosphere of HAT-P-7b. We also analyze the eight transits of HAT-P-7b present in the EPOXI data set and improve the constraints on the system parameters, finding a period of P = 2.2047308 +- 0.0000025 days, a stellar radius of R{sub *} = 1.824 +- 0.089 R{sub sun}, a planetary radius of R{sub p} = 1.342 +- 0.068 R{sub Jup}, and an inclination of i = 85.7{sup +3.5}{sub -2.2} deg.

  8. Reconstruction of BL7B for UV, VIS and IR spectroscopy with a 3 m normal-incidence monochromator.

    PubMed

    Fukui, K; Nakagawa, H; Shimoyama, I; Nakagawa, K; Okamura, H; Nanba, T; Hasumoto, M; Kinoshita, T

    1998-05-01

    The beamline BL7B at the UVSOR facility for solid-state spectroscopy is currently under reconstruction. This reconstruction mainly involves the replacement of the 1 m Seya-Namioka-type monochromator (50-600 nm) with a 3 m NIM (modified version of McPherson model 2253), which covers the 50-1000 nm range with three gratings. The deviation angle of the gratings is 15 degrees. For linear and circular polarization experiments, the beamline optics consist of a two-grazing-incidence (87.5 degrees ) pre-mirror system and a normal-incidence (15 degrees ) post-mirror.

  9. Emission behavior of sudan red 7B on dogbone-shaped gold nanorods: Aspect ratio dependence of the metallic nanostructures

    NASA Astrophysics Data System (ADS)

    Rahman, Dewan S.; Sharma, Debdulal; Ghosh, Sujit Kumar

    2014-01-01

    Cetyltrimethylammonium bromide-stabilized 'dogbone-shaped' gold nanorods of aspect ratio varying from 1 to 6 have been synthesized by seed-mediated growth method in aqueous medium. Then, sudan red 7B, an alien molecular probe has been used as local probe to elucidate aspect ratio dependence of the nanorods on the photophysical properties of the dye molecules. It is seen that the relative intensity decreases exponentially with increasing aspect ratio and has been attributed to decrease in overall surface area for a particular concentration of the nanorods. The mechanism of fluorescence quenching has been ascribed to the electron and energy transfer processes in the gold-fluorophore hybrid nanostructures.

  10. EOS7C Version 1.0 TOUGH2 Module for Carbon Dioxide or Nitrogen in Natural Gas

    2008-01-11

    EOS7C is a TOUGH2 module for multicomponent gas mixtures in the systems methane-carbon dioxide (CH{sub 4}-CO{sub 2}) or methane-nitrogen (CH{sub 4}-N{sub 2}) with or without an aqueous phase and H{sub 2}O vapor. EOS7C uses a cubic equation of state and an accurate solubility formulation along with a multiphase Darcy's Law to model flow and transport of gas and aqueous phase mixtures over a wide range of pressures and temperatures appropriate to subsurface geologic carbon sequestrationmore » sites and natural gas reservoirs. EOS7C models supercritical CO{sub 2{ and subcritical CO{sub 2} as a non-condensible gas, hence EOS7C does not model the transitions to liquid or solid CO{sub 2} conditions. The components modeled in EOS7C are water, brine, non-condensible gas, gas tracer, methane, and optional heat. The non-condensible gas (NCG) can be selected by the user to be CO{sub 2} or N{sub 2}. The real gas properties module has options for Peng-Robinson, Redlich-Kwong, or Soave-Redlich-Kwong equations of state to calculate gas mixture density, enthalpy departure, and viscosity. Partitioning of the NCG and CH{sub 4} between the aqueous and gas phases is calculated using a very accurate chemical equilibrium approach. Transport of the gaseous and dissolved components is by advection and Fickian molecular diffusion. EOS7C is written in FORTAN77.« less

  11. Apical targeting and Golgi retention signals reside within a 9-amino acid sequence in the copper-ATPase, ATP7B.

    PubMed

    Braiterman, Lelita; Nyasae, Lydia; Guo, Yan; Bustos, Rodrigo; Lutsenko, Svetlana; Hubbard, Ann

    2009-02-01

    ATP7B is a copper-transporting P-type ATPase present predominantly in liver. In basal copper, hepatic ATP7B is in a post-trans-Golgi network (TGN) compartment where it loads cytoplasmic Cu(I) onto newly synthesized ceruloplasmin. When copper levels rise, the protein redistributes via unique vesicles to the apical periphery where it exports intracellular Cu(I) into bile. We want to understand the mechanisms regulating the copper-sensitive trafficking of ATP7B. Earlier, our laboratory reported the presence of apical targeting/TGN retention information within residues 1-63 of human ATP7B; deletion of these residues resulted in a mutant protein that was not efficiently retained in the post-TGN in low copper and constitutively trafficked to the basolateral membrane of polarized, hepatic WIF-B cells with and without copper (13). In this study, we used mutagenesis and adenovirus infection of WIF-B cells followed by confocal immunofluorescence microscopy analysis to identify the precise retention/targeting sequences in the context of full-length ATP7B. We also analyzed the expression of selected mutants in livers of copper-deficient and -loaded mice. Our combined results clearly demonstrate that nine amino acids, F(37)AFDNVGYE(45), comprise an essential apical targeting determinant for ATP7B in elevated copper and participate in the TGN retention of the protein under low-copper conditions. The signal is novel, does not require phosphorylation, and is highly conserved in approximately 24 species of ATP7B. Furthermore, N41S, which is part of the signal we identified, is the first and only Wilson disease-causing missense mutation in residues 1-63 of ATP7B. Expression of N41S-ATP7B in WIF-B cells severely disabled the targeting and retention of the protein. We present a working model of how this physiologically relevant signal might work. PMID:19033537

  12. INVESTIGATION OF SYSTEMATIC EFFECTS IN KEPLER DATA: SEASONAL VARIATIONS IN THE LIGHT CURVE OF HAT-P-7b

    SciTech Connect

    Van Eylen, V.; Lindholm Nielsen, M.; Hinrup, B.; Tingley, B.; Kjeldsen, H.

    2013-09-10

    With years of Kepler data currently available, the measurement of variations in planetary transit depths over time can now be attempted. To do so, it is of primary importance to understand which systematic effects may affect the measurement of transits. We aim to measure the stability of Kepler measurements over years of observations. We present a study of the depth of about 500 transit events of the Hot Jupiter HAT-P-7b, using 14 quarters (Q0-Q13) of data from the Kepler satellite. We find a systematic variation in the depth of the primary transit, related to quarters of data and recurring yearly. These seasonal variations are about 1%. Within seasons, we find no evidence for trends. We speculate that the cause of the seasonal variations could be unknown field crowding or instrumental artifacts. Our results show that care must be taken when combining transits throughout different quarters of Kepler data. Measuring the relative planetary radius of HAT-P-7b without taking these systematic effects into account leads to unrealistically low error estimates. This effect could be present in all Kepler targets. If so, relative radius measurements of all Hot Jupiters to a precision much better than 1% are unrealistic.

  13. ATMOSPHERE AND SPECTRAL MODELS OF THE KEPLER-FIELD PLANETS HAT-P-7b AND TrES-2

    SciTech Connect

    Spiegel, David S.; Burrows, Adam E-mail: burrows@astro.princeton.ed

    2010-10-10

    We develop atmosphere models of two of the three Kepler-field planets that were known prior to the start of the Kepler mission (HAT-P-7b and TrES-2). We find that published Kepler and Spitzer data for HAT-P-7b appear to require an extremely hot upper atmosphere on the dayside, with a strong thermal inversion and little day-night redistribution. The Spitzer data for TrES-2 suggest a mild thermal inversion with moderate day-night redistribution. We examine the effect of nonequilibrium chemistry on TrES-2 model atmospheres and find that methane levels must be adjusted by extreme amounts in order to cause even mild changes in atmospheric structure and emergent spectra. Our best-fit models to the Spitzer data for TrES-2 lead us to predict a low secondary eclipse planet-star flux ratio ({approx}<2 x 10{sup -5}) in the Kepler bandpass, which is consistent with what very recent observations have found. Finally, we consider how the Kepler-band optical flux from a hot exoplanet depends on the strength of a possible extra optical absorber in the upper atmosphere. We find that the optical flux is not monotonic in optical opacity, and the non-monotonicity is greater for brighter, hotter stars.

  14. Polyamines release the let-7b-mediated suppression of initiation codon recognition during the protein synthesis of EXT2

    PubMed Central

    Imamura, Masataka; Higashi, Kyohei; Yamaguchi, Katsutoshi; Asakura, Kiryu; Furihata, Tomomi; Terui, Yusuke; Satake, Toshihiko; Maegawa, Jiro; Yasumura, Kazunori; Ibuki, Ai; Akase, Tomoko; Nishimura, Kazuhiro; Kashiwagi, Keiko; Linhardt, Robert J.; Igarashi, Kazuei; Toida, Toshihiko

    2016-01-01

    Proteoglycans (PGs), a family of glycosaminoglycan (GAG)-protein glycoconjugates, contribute to animal physiology through interactions between their glycan chains and growth factors, chemokines and adhesion molecules. However, it remains unclear how GAG structures are changed during the aging process. Here, we found that polyamine levels are correlated with the expression level of heparan sulfate (HS) in human skin. In cultured cell lines, the EXT1 and EXT2 enzymes, initiating HS biosynthesis, were stimulated at the translational level by polyamines. Interestingly, the initiation codon recognition by 43S preinitiation complex during EXT2 translation is suppressed by let-7b, a member of the let-7 microRNA family, through binding at the N-terminal amino acid coding sequence in EXT2 mRNA. Let-7b-mediated suppression of initiation codon depends on the length of 5′-UTR of EXT2 mRNA and its suppression is inhibited in the presence of polyamines. These findings provide new insights into the HS biosynthesis related to miRNA and polyamines. PMID:27650265

  15. Polyamines release the let-7b-mediated suppression of initiation codon recognition during the protein synthesis of EXT2.

    PubMed

    Imamura, Masataka; Higashi, Kyohei; Yamaguchi, Katsutoshi; Asakura, Kiryu; Furihata, Tomomi; Terui, Yusuke; Satake, Toshihiko; Maegawa, Jiro; Yasumura, Kazunori; Ibuki, Ai; Akase, Tomoko; Nishimura, Kazuhiro; Kashiwagi, Keiko; Linhardt, Robert J; Igarashi, Kazuei; Toida, Toshihiko

    2016-01-01

    Proteoglycans (PGs), a family of glycosaminoglycan (GAG)-protein glycoconjugates, contribute to animal physiology through interactions between their glycan chains and growth factors, chemokines and adhesion molecules. However, it remains unclear how GAG structures are changed during the aging process. Here, we found that polyamine levels are correlated with the expression level of heparan sulfate (HS) in human skin. In cultured cell lines, the EXT1 and EXT2 enzymes, initiating HS biosynthesis, were stimulated at the translational level by polyamines. Interestingly, the initiation codon recognition by 43S preinitiation complex during EXT2 translation is suppressed by let-7b, a member of the let-7 microRNA family, through binding at the N-terminal amino acid coding sequence in EXT2 mRNA. Let-7b-mediated suppression of initiation codon depends on the length of 5'-UTR of EXT2 mRNA and its suppression is inhibited in the presence of polyamines. These findings provide new insights into the HS biosynthesis related to miRNA and polyamines. PMID:27650265

  16. Non-classical heteroacenes: synthesis and properties of anthra[2,3-c:6,7-c']dithiophene derivatives.

    PubMed

    Djukic, Brandon; Perepichka, Dmitrii F

    2011-12-21

    A new class of linear heteroacenes, anthra[2,3-c:6,7-c']dithiophenes, containing non-classical (non-Kekulé) thiophene in a 22-electron π-conjugated system have been prepared by electrochemical reduction. The electronic properties have been studied by DFT calculations and spectroelectrochemistry, supporting the formation of a triplet electronic structure. PMID:22042513

  17. The 3D structure of the defense-related rice protein Pir7b predicted by homology modeling and ligand binding studies.

    PubMed

    Luo, Quan; Han, Wei-Wei; Zhou, Yi-Han; Yao, Yuan; Li, Ze-Sheng

    2008-07-01

    To better understand the ligand-binding mechanism of protein Pir7b, important part in detoxification of a pathogen-derived compound against Pyricularia oryzae, a 3D structure model of protein Pir7b was constructed based on the structure of the template SABP2. Three substrates were docking to this protein, two of them were proved to be active, and some critical residues are identified, which had not been confirmed by the experiments. His87 and Leu17 considered as 'oxyanion hole' contribute to initiating the Ser86 nucleophilic attack. Gln187 and Asp139 can form hydrogen bonds with the anilid group to maintain the active binding orientation with the substrates. The docking model can well interpret the specificity of protein Pir7b towards the anilid moiety of the substrates and provide valuable structure information about the ligand binding to protein Pir7b. PMID:18449577

  18. Computational Investigations of Trichoderma Reesei Cel7A Suggest New Routes for Enzyme Activity Improvements

    SciTech Connect

    Beckham, G. T.; Payne, C. M.; Bu, L.; Taylor, C. B.; McCabe, C.; Chu, J. W.; Himmel, M. E.; Crowley, M. F.

    2012-01-01

    The Trichoderma reesei Family 7 cellulase (Cel7A) is a key industrial enzyme in the production of biofuels from lignocellulosic biomass. It is a multi-modular enzyme with a Family 1 carbohydrate-binding module, a flexible O-glycosylated linker, and a large catalytic domain. We have used simulation to elucidate new functions for the 3 sub-domains, which suggests new routes to increase the activity of this central enzyme. These findings include new roles for glycosylation, which we have shown can be used to tune the binding affinity. We have also examined the structures of the catalytically-active complex of Cel7A and its non-processive counterpart, Cel7B, engaged on cellulose, which suggests allosteric mechanisms involved in chain binding when these cellulases are complexed on cellulose. Our computational results also suggest that product inhibition varies significantly between Cel7A and Cel7B, and we offer a molecular-level explanation for this observation. Finally, we discuss simulations of the absolute and relative binding free energy of cellulose ligands and various mutations along the CD tunnel, which will affect processivity and the ability of Cel7A (and related enzymes) to digest cellulose. These results highlight new considerations in protein engineering for processive and non-processive cellulases for production of lignocellulosic biofuels.

  19. Interactions between Copper-binding Sites Determine the Redox Status and Conformation of the Regulatory N-terminal Domain of ATP7B*

    PubMed Central

    LeShane, Erik S.; Shinde, Ujwal; Walker, Joel M.; Barry, Amanda N.; Blackburn, Ninian J.; Ralle, Martina; Lutsenko, Svetlana

    2010-01-01

    Copper-transporting ATPase ATP7B is essential for human copper homeostasis and normal liver function. ATP7B has six N-terminal metal-binding domains (MBDs) that sense cytosolic copper levels and regulate ATP7B. The mechanism of copper sensing and signal integration from multiple MBDs is poorly understood. We show that MBDs communicate and that this communication determines the oxidation state and conformation of the entire N-terminal domain of ATP7B (N-ATP7B). Mutations of copper-coordinating Cys to Ala in any MBD (2, 3, 4, or 6) change the N-ATP7B conformation and have distinct functional consequences. Mutating MBD2 or MBD3 causes Cys oxidation in other MBDs and loss of copper binding. In contrast, mutation of MBD4 and MBD6 does not alter the redox status and function of other sites. Our results suggest that MBD2 and MBD3 work together to regulate access to other metal-binding sites, whereas MBD4 and MBD6 receive copper independently, downstream of MBD2 and MBD3. Unlike Ala substitutions, the Cys-to-Ser mutation in MBD2 preserves the conformation and reduced state of N-ATP7B, suggesting that hydrogen bonds contribute to interdomain communications. Tight coupling between MBDs suggests a mechanism by which small changes in individual sites (induced by copper binding or mutation) result in stabilization of distinct conformations of the entire N-ATP7B and altered exposure of sites for interactions with regulatory proteins. PMID:20032459

  20. Communication between the N and C termini is required for copper-stimulated Ser/Thr phosphorylation of Cu(I)-ATPase (ATP7B).

    PubMed

    Braiterman, Lelita T; Gupta, Arnab; Chaerkady, Raghothama; Cole, Robert N; Hubbard, Ann L

    2015-04-01

    The Wilson disease protein ATP7B exhibits copper-dependent trafficking. In high copper, ATP7B exits the trans-Golgi network and moves to the apical domain of hepatocytes where it facilitates elimination of excess copper through the bile. Copper levels also affect ATP7B phosphorylation. ATP7B is basally phosphorylated in low copper and becomes more phosphorylated ("hyperphosphorylated") in elevated copper. The functional significance of hyperphosphorylation remains unclear. We showed that hyperphosphorylation occurs even when ATP7B is restricted to the trans-Golgi network. We performed comprehensive phosphoproteomics of ATP7B in low versus high copper, which revealed that 24 Ser/Thr residues in ATP7B could be phosphorylated, and only four of these were copper-responsive. Most of the phosphorylated sites were found in the N- and C-terminal cytoplasmic domains. Using truncation and mutagenesis, we showed that inactivation or elimination of all six N-terminal metal binding domains did not block copper-dependent, reversible, apical trafficking but did block hyperphosphorylation in hepatic cells. We showed that nine of 15 Ser/Thr residues in the C-terminal domain were phosphorylated. Inactivation of 13 C-terminal phosphorylation sites reduced basal phosphorylation and eliminated hyperphosphorylation, suggesting that copper binding at the N terminus propagates to the ATP7B C-terminal region. C-terminal mutants with either inactivating or phosphomimetic substitutions showed little effect upon copper-stimulated trafficking, indicating that trafficking does not depend on phosphorylation at these sites. Thus, our studies revealed that copper-dependent conformational changes in the N-terminal region lead to hyperphosphorylation at C-terminal sites, which seem not to affect trafficking and may instead fine-tune copper sequestration. PMID:25666620

  1. Tissue-specific expression of cell-surface Qa-2 antigen from a transfected Q7b gene of C57BL/10 mice

    PubMed Central

    1987-01-01

    We screened a cDNA library prepared from a BALB.B10 CTL clone that expresses Qa-2 antigen, and isolated four clones derived from Q7b, a Qa region gene of C57BL/10. One of these Q7b cDNAs and the Q7b chromosomal gene were subcloned into expression vectors and transfected into L cells and R1.1 thymoma cells. We found that the chromosomal Q7b gene expresses Qa-2 on the surface of R1.1 cells, but not on L cells while the Q7b cDNA expresses protein on the surface of both cell types. The levels of Qa-2 expression do not correlate with the total levels of Q7b mRNA in these transfectants. Our results suggest that the tissue- specific expression of Qa-2 may be controlled, in part, by mechanisms of alternate RNA splicing. By using hybrid gene constructs, we have mapped the tissue-specific element to the 3' part of the gene, downstream of a site near the middle of exon 4. The hybrid polypeptides differ significantly in their transmembrane and cytoplasmic regions. These portions of the protein also may play a role in the tissue- specific expression of Qa-2. PMID:3502706

  2. miRNA let-7b modulates macrophage polarization and enhances tumor-associated macrophages to promote angiogenesis and mobility in prostate cancer.

    PubMed

    Wang, Zhigang; Xu, Lu; Hu, Yinying; Huang, Yanqin; Zhang, Yujuan; Zheng, Xiufen; Wang, Shanshan; Wang, Yifan; Yu, Yanrong; Zhang, Meng; Yuan, Keng; Min, Weiping

    2016-05-09

    Macrophage polarization is a highly plastic physiological process that responds to a variety of environmental factors by changing macrophage phenotype and function. Tumor-associated macrophages (TAMs) are generally recognized as promoting tumor progression. As universal regulators, microRNAs (miRNAs) are functionally involved in numerous critical cellular processes including macrophage polarization. Let-7b, a miRNA, has differential expression patterns in inflamed tissues compared with healthy controls. However, whether and how miRNA let-7b regulates macrophage phenotype and function is unclear. In this report, we find that up-regulation of let-7b is characteristic of prostatic TAMs, and down-regulation of let-7b in TAMs leads to changes in expression profiles of inflammatory cytokines, such as IL-12, IL-23, IL-10 and TNF-α. As a result, TAMs treated with let-7b inhibitors reduce angiogenesis and prostate carcinoma (PCa) cell mobility. Let-7b may play a vital role in regulating macrophage polarization, thus modulating the prognosis of prostate cancer.

  3. The discrepancy between the absence of copper deposition and the presence of neuronal damage in the brain of Atp7b(-/-) mice.

    PubMed

    Dong, Yi; Shi, Sheng-Sheng; Chen, Sheng; Ni, Wang; Zhu, Min; Wu, Zhi-Ying

    2015-02-01

    Wilson's disease (WD) is caused by mutations within the copper-transporting ATPase (ATP7B), characterized by copper deposition in various organs, principally the liver and the brain. With the availability of Atp7b(-/-) mice, the valid animal model of WD, the mechanism underlying copper-induced hepatocyte necrosis has been well understood. Nonetheless, little is known about the adverse impact of copper accumulation on the brain in WD. Therefore, the aim of this study was to identify copper disturbances according to various brain compartments and further dissect the causal relationship between copper storage and neuronal damage using Atp7b(-/-) mice. Copper levels in the liver, whole brain, brain compartments and basal ganglia mitochondria of Atp7b(-/-) mice and age-matched controls were measured by atomic absorption spectroscopy. Delicate electron microscopic studies on hepatocytes and neurons in the basal ganglia were performed. Here we further confirmed the remarkably elevated copper content and abnormal ultrastructure findings in livers of Atp7b(-/-) mice. Interestingly, we found the ultrastructure abnormalities in neurons of the basal ganglia of Atp7b(-/-) mice, whereas copper deposition was not detected in the whole brain, even within the basal ganglia and its mitochondria. The disparity provided a new understanding of neuronal dysfunction in WD, and strongly indicated that copper might not be the sole causative player and other unidentified pathogenic factors could enhance the toxic effects of copper on neurons in WD.

  4. miRNA let-7b modulates macrophage polarization and enhances tumor-associated macrophages to promote angiogenesis and mobility in prostate cancer

    PubMed Central

    Wang, Zhigang; Xu, Lu; Hu, Yinying; Huang, Yanqin; Zhang, Yujuan; Zheng, Xiufen; Wang, Shanshan; Wang, Yifan; Yu, Yanrong; Zhang, Meng; Yuan, Keng; Min, Weiping

    2016-01-01

    Macrophage polarization is a highly plastic physiological process that responds to a variety of environmental factors by changing macrophage phenotype and function. Tumor-associated macrophages (TAMs) are generally recognized as promoting tumor progression. As universal regulators, microRNAs (miRNAs) are functionally involved in numerous critical cellular processes including macrophage polarization. Let-7b, a miRNA, has differential expression patterns in inflamed tissues compared with healthy controls. However, whether and how miRNA let-7b regulates macrophage phenotype and function is unclear. In this report, we find that up-regulation of let-7b is characteristic of prostatic TAMs, and down-regulation of let-7b in TAMs leads to changes in expression profiles of inflammatory cytokines, such as IL-12, IL-23, IL-10 and TNF-α. As a result, TAMs treated with let-7b inhibitors reduce angiogenesis and prostate carcinoma (PCa) cell mobility. Let-7b may play a vital role in regulating macrophage polarization, thus modulating the prognosis of prostate cancer. PMID:27157642

  5. Structural and metabolic changes in Atp7b-/- mouse liver and potential for new interventions in Wilson's disease.

    PubMed

    Huster, Dominik

    2014-05-01

    Wilson's disease (WD) is caused by ATP7B mutations and results in copper accumulation and toxicity in liver and brain tissues. The specific mechanisms underlying copper toxicity are still poorly understood. Mouse models have revealed new insights into pathomechanisms of hepatic WD. Mitochondrial damage is observed in livers of WD patients and in mouse models; copper induces fragmentation of mitochondrial membrane lipids, particularly cardiolipin, with deleterious effects on both mitochondrial integrity and function. Copper accumulation also induces chronic inflammation in WD livers, which is followed by regeneration in parts of the liver and occasionally neoplastic proliferation. Gene expression studies using microarrays have aided our understanding of the molecular basis of these changes. Copper overload alters cholesterol biosynthesis in hepatocytes resulting in reduced liver and serum cholesterol. Experiments are currently underway to elucidate the link between copper and cholesterol metabolism. These findings may facilitate the development of specific therapies to ameliorate WD progression.

  6. KELT-7b: A Hot Jupiter Transiting a Bright V = 8.54 Rapidly Rotating F-star

    NASA Astrophysics Data System (ADS)

    Bieryla, Allyson; Collins, Karen; Beatty, Thomas G.; Eastman, Jason; Siverd, Robert J.; Pepper, Joshua; Gaudi, B. Scott; Stassun, Keivan G.; Cañas, Caleb; Latham, David W.; Buchhave, Lars A.; Sanchis-Ojeda, Roberto; Winn, Joshua N.; Jensen, Eric L. N.; Kielkopf, John F.; McLeod, Kim K.; Gregorio, Joao; Colón, Knicole D.; Street, Rachel; Ross, Rachel; Penny, Matthew; Mellon, Samuel N.; Oberst, Thomas E.; Fulton, Benjamin J.; Wang, Ji; Berlind, Perry; Calkins, Michael L.; Esquerdo, Gilbert A.; DePoy, Darren L.; Gould, Andrew; Marshall, Jennifer; Pogge, Richard; Trueblood, Mark; Trueblood, Patricia

    2015-07-01

    We report the discovery of KELT-7b, a transiting hot Jupiter with a mass of 1.28+/- 0.18 MJ, radius of {1.533}-0.047+0.046 RJ, and an orbital period of 2.7347749+/- 0.0000039 days. The bright host star (HD 33643; KELT-7) is an F-star with V = 8.54, Teff = 6789{}-49+50 K, [Fe/H] = {0.139}-0.081+0.075, and {log}\\{\\text{}}g\\=4.149+/- 0.019. It has a mass of {1.535}-0.054+0.066 M⊙, a radius of {1.732}-0.045+0.043 R⊙, and is the fifth most massive, fifth hottest, and the ninth brightest star known to host a transiting planet. It is also the brightest star around which Kilodegree Extremely Little Telescope (KELT) has discovered a transiting planet. Thus, KELT-7b is an ideal target for detailed characterization given its relatively low surface gravity, high equilibrium temperature, and bright host star. The rapid rotation of the star (73+/- 0.5 km s‑1) results in a Rossiter–McLaughlin effect with an unusually large amplitude of several hundred m s‑1. We find that the orbit normal of the planet is likely to be well-aligned with the stellar spin axis, with a projected spin–orbit alignment of λ = 9.°7 ± 5.°2. This is currently the second most rapidly rotating star to have a reflex signal (and thus mass determination) due to a planetary companion measured.

  7. KELT-7b: A Hot Jupiter Transiting a Bright V = 8.54 Rapidly Rotating F-star

    NASA Astrophysics Data System (ADS)

    Bieryla, Allyson; Collins, Karen; Beatty, Thomas G.; Eastman, Jason; Siverd, Robert J.; Pepper, Joshua; Gaudi, B. Scott; Stassun, Keivan G.; Cañas, Caleb; Latham, David W.; Buchhave, Lars A.; Sanchis-Ojeda, Roberto; Winn, Joshua N.; Jensen, Eric L. N.; Kielkopf, John F.; McLeod, Kim K.; Gregorio, Joao; Colón, Knicole D.; Street, Rachel; Ross, Rachel; Penny, Matthew; Mellon, Samuel N.; Oberst, Thomas E.; Fulton, Benjamin J.; Wang, Ji; Berlind, Perry; Calkins, Michael L.; Esquerdo, Gilbert A.; DePoy, Darren L.; Gould, Andrew; Marshall, Jennifer; Pogge, Richard; Trueblood, Mark; Trueblood, Patricia

    2015-07-01

    We report the discovery of KELT-7b, a transiting hot Jupiter with a mass of 1.28+/- 0.18 MJ, radius of {1.533}-0.047+0.046 RJ, and an orbital period of 2.7347749+/- 0.0000039 days. The bright host star (HD 33643; KELT-7) is an F-star with V = 8.54, Teff = 6789{}-49+50 K, [Fe/H] = {0.139}-0.081+0.075, and {log}\\{\\text{}}g\\=4.149+/- 0.019. It has a mass of {1.535}-0.054+0.066 M⊙, a radius of {1.732}-0.045+0.043 R⊙, and is the fifth most massive, fifth hottest, and the ninth brightest star known to host a transiting planet. It is also the brightest star around which Kilodegree Extremely Little Telescope (KELT) has discovered a transiting planet. Thus, KELT-7b is an ideal target for detailed characterization given its relatively low surface gravity, high equilibrium temperature, and bright host star. The rapid rotation of the star (73+/- 0.5 km s-1) results in a Rossiter-McLaughlin effect with an unusually large amplitude of several hundred m s-1. We find that the orbit normal of the planet is likely to be well-aligned with the stellar spin axis, with a projected spin-orbit alignment of λ = 9.°7 ± 5.°2. This is currently the second most rapidly rotating star to have a reflex signal (and thus mass determination) due to a planetary companion measured.

  8. Spectral Line Survey toward RCrA IRS7B in the 345 GHz Window with ASTE

    NASA Astrophysics Data System (ADS)

    Watanabe, Y.; Sakai, N.; Lindberg, J.; Jorgensen, J.; Bisschopi, S.; Yamamoto, S.

    2011-05-01

    Recently, it has been pointed out that the chemical composition in the low-mass protostars varies from source to source. Two extreme cases are the hot corinos where various complex organic molecules are abundant, and the WCCC (Warm Carbon-Chain Chemistry) sources where carbon-chain molecules are abundant. Understanding an origin of such chemical diversity is an important target for astrochemistry. We have conducted a spectral line survey in the 332 - 364 GHz band (figure 1) with ASTE toward a low mass class 0 protostar in the Corona Australis (R CrA IRS7B), which have been recognized as a hot corino candidate because of its bright emissions of H_2CO and CH_3OH. We have also performed a supplementary observations in the 450 GHz band. In total, 16 molecular species including CO, CS, CN, NO, CCH, c-C3H2, HCO+, H2CO and CH3OH are identified. Strong emissions of CN, and CCH are observed, whereas, complex organic molecules and long carbon-chain molecules were not detected. The rotation temperature of CH3OH is evaluated to be ˜ 16 K, which is much lower than that in the typical hot corino IRAS 16293-2422 (˜85 K). The deuterium fractionation ratio for CCH and H2CO are obtained to be 0.06-0.01, which are an order of magnitude lower than that found in the hot corino. Furthermore c-C3H2, whose production pathway is related to carbon-chain molecules, is abundant, and its rotation temperature is similar to that of CH3OH. From these results, it is likely that the RCrA IRS7B is a borderline source between the hot corinos and the WCCC sources. A related survey in the 230 GHz band is in progress for full characterization of the chemical composition of this low-mass protostellar source (Lindberg et al. 2011).

  9. Identification and Phylogenetic Analysis of a CC-NBS-LRR Encoding Gene Assigned on Chromosome 7B of Wheat

    PubMed Central

    Gong, Caiyan; Cao, Shuanghe; Fan, Renchun; Wei, Bo; Chen, Guiping; Wang, Xianping; Li, Yiwen; Zhang, Xiangqi

    2013-01-01

    Hexaploid wheat displays limited genetic variation. As a direct A and B genome donor of hexaploid wheat, tetraploid wheat represents an important gene pool for cultivated bread wheat. Many disease resistant genes express conserved domains of the nucleotide-binding site and leucine-rich repeats (NBS-LRR). In this study, we isolated a CC-NBS-LRR gene locating on chromosome 7B from durum wheat variety Italy 363, and designated it TdRGA-7Ba. Its open reading frame was 4014 bp, encoding a 1337 amino acid protein with a complete NBS domain and 18 LRR repeats, sharing 44.7% identity with the PM3B protein. TdRGA-7Ba expression was continuously seen at low levels and was highest in leaves. TdRGA-7Ba has another allele TdRGA-7Bb with a 4 bp deletion at position +1892 in other cultivars of tetraploid wheat. In Ae. speltoides, as a B genome progenitor, both TdRGA-7Ba and TdRGA-7Bb were detected. In all six species of hexaploid wheats (AABBDD), only TdRGA-7Bb existed. Phylogenic analysis showed that all TdRGA-7Bb type genes were grouped in one sub-branch. We speculate that TdRGA-7Bb was derived from a TdRGA-7Ba mutation, and it happened in Ae. speltoides. Both types of TdRGA-7B participated in tetraploid wheat formation. However, only the TdRGA-7Bb was retained in hexaploid wheat. PMID:23887654

  10. Molecular characterization and bioinformatics analysis of Ncoa7B, a novel ovulation-associated and reproduction system-specific Ncoa7 isoform.

    PubMed

    Shkolnik, Ketty; Ben-Dor, Shifra; Galiani, Dalia; Hourvitz, Ariel; Dekel, Nava

    2008-03-01

    In the present work, we employed bioinformatics search tools to select ovulation-associated cDNA clones with a preference for those representing putative novel genes. Detailed characterization of one of these transcripts, 6C3, by real-time PCR and RACE analyses led to identification of a novel ovulation-associated gene, designated Ncoa7B. This gene was found to exhibit a significant homology to the Ncoa7 gene that encodes a conserved tissue-specific nuclear receptor coactivator. Unlike Ncoa7, Ncoa7B possesses a unique and highly conserved exon at the 5' end and encodes a protein with a unique N-terminal sequence. Extensive bioinformatics analysis has revealed that Ncoa7B has one identifiable domain, TLDc, which has recently been suggested to be involved in protection from oxidative DNA damage. An alignment of TLDc domain containing proteins was performed, and the closest relative identified was OXR1, which also has a corresponding, highly related short isoform, with just a TLDc domain. Moreover, Ncoa7B expression, as seen to date, seems to be restricted to mammals, while other TLDc family members have no such restriction. Multiple tissue analysis revealed that unlike Ncoa7, which was abundant in a variety of tissues with the highest expression in the brain, Ncoa7B mRNA expression is restricted to the reproductive system organs, particularly the uterus and the ovary. The ovarian expression of Ncoa7B was stimulated by human chorionic gonadotropin. Additionally, using real-time PCR, we demonstrated the involvement of multiple signaling pathways for Ncoa7B expression on preovulatory follicles. PMID:18299425

  11. cAMP/PKA/CREB/GLT1 signaling involved in the antidepressant-like effects of phosphodiesterase 4D inhibitor (GEBR-7b) in rats

    PubMed Central

    Liu, Xu; Guo, Haibiao; Sayed, Mohammad Daud SOM; Lu, Yang; Yang, Ting; Zhou, Dongsheng; Chen, Zhongming; Wang, Haitao; Wang, Chuang; Xu, Jiangping

    2016-01-01

    Objectives GEBR-7b, a potential phosphodiesterase 4D inhibitor, has been shown to have memory-enhancing effects in rodents. However, it is still unknown whether GEBR-7b also has the antidepressant-like effects in rats. Herein, we examined the potential of GEBR-7b to attenuate depression-like behaviors in the rat model of depression induced by chronic unpredictable stress (CUS). Next, we also investigated the alterations of cyclic adenosine monophosphate (cAMP), protein kinase A (PKA) catalytic subunit (PKAca), cAMP response element-binding (CREB), and glutamate transporter 1 (GLT1) levels produced by GEBR-7b in the rats model of depression. Methods Effects of GEBR-7b on CUS (35 days)-induced depression-like behaviors were examined by measuring immobility time in the forced swimming test (FST). Hippocampal cAMP levels were examined by enzyme-linked immunosorbent assay, whereas PKAca, phosphorylation of CREB (pCREB), CREB, and GLT1 in the hippocampus of rats were subjected to Western blot analysis. Results CUS exposure caused a depression-like behavior evidenced by the increased immobility time in FST. Depression-like behavior induced by CUS was accompanied by a significant increased GLT, decreased cAMP, PKAca, pCREB activities in hippocampus. However, repeated GEBR-7b administration significantly reversed CUS-induced depression-like behavior and changes of cAMP/PKA/CREB/GLT1 signaling. No alteration was observed in locomotor activity in open field test. Conclusion These findings indicate that GEBR-7b reversed the depression-like behaviors induced by CUS in rats, which is at least in part mediated by modulating cAMP, PKAca, pCREB, and GLT1 levels in the hippocampus of rats, supporting its neuroprotective potential against behavioral and biochemical dysfunctions induced by CUS. PMID:26855578

  12. Membrane Docking of the Synaptotagmin 7 C2A Domain: Computation Reveals Interplay between Electrostatic and Hydrophobic Contributions.

    PubMed

    Chon, Nara Lee; Osterberg, J Ryan; Henderson, Jack; Khan, Hanif M; Reuter, Nathalie; Knight, Jefferson D; Lin, Hai

    2015-09-22

    The C2A domain of synaptotagmin 7 (Syt7) is a Ca(2+) and membrane binding module that docks and inserts into cellular membranes in response to elevated intracellular Ca(2+) concentrations. Like other C2 domains, Syt7 C2A binds Ca(2+) and membranes primarily through three loop regions; however, it docks at Ca(2+) concentrations much lower than those required for other Syt C2A domains. To probe structural components of its unusually strong membrane docking, we conducted atomistic molecular dynamics simulations of Syt7 C2A under three conditions: in aqueous solution, in the proximity of a lipid bilayer membrane, and embedded in the membrane. The simulations of membrane-free protein indicate that Syt7 C2A likely binds three Ca(2+) ions in aqueous solution, consistent with prior experimental reports. Upon membrane docking, the outermost Ca(2+) ion interacts directly with lipid headgroups, while the other two Ca(2+) ions remain chelated by the protein. The membrane-bound domain was observed to exhibit large-amplitude swinging motions relative to the membrane surface, varying by up to 70° between a more parallel and a more perpendicular orientation, both during and after insertion of the Ca(2+) binding loops into the membrane. The computed orientation of the membrane-bound protein correlates well with experimental electron paramagnetic resonance measurements presented in the preceding paper ( DOI: 10.1021/acs.biochem.5b00421 ). In particular, the strictly conserved residue Phe229 inserted stably ∼4 Å below the average depth of lipid phosphate groups, providing critical hydrophobic interactions anchoring the domain in the membrane. Overall, the position and orientation of Syt7 C2A with respect to the membrane are consistent with experiments.

  13. First principles study on the electronic structures and stability of Cr 7C 3 type multi-component carbides

    NASA Astrophysics Data System (ADS)

    Xiao, B.; Feng, J.; Zhou, C. T.; Xing, J. D.; Xie, X. J.; Chen, Y. H.

    2008-06-01

    First principles calculations were conducted to investigate the stabilities of six multi-component carbides of Cr 7C 3 by calculating the cohesive energy and formation enthalpy of them. The theoretical predictions were compared with the experimental results and they were in agreement with each other. The electronic structures of the six carbides were also calculated in order to provide more information about the relationship between the stability and crystal compositions at atomic scale.

  14. Salinity-dependent copper accumulation in the guppy Poecilia vivipara is associated with CTR1 and ATP7B transcriptional regulation.

    PubMed

    da Silva, Evelise Sampaio; Abril, Sandra Isabel Moreno; Zanette, Juliano; Bianchini, Adalto

    2014-07-01

    Copper (Cu) accumulation and regulation of key-genes involved in Cu homeostasis were evaluated in freshwater- and saltwater-acclimated guppies Poecilia vivipara. Fish were exposed (96h) to environmentally relevant concentrations of dissolved Cu (0, 5.0, 9.0 and 20.0μg/L). In freshwater guppies, gill and liver Cu accumulation was dependent on Cu concentration in the exposure medium. In saltwater guppies, this dependence was observed only in the gut. These findings indicate that Cu accumulation was salinity- and tissue-dependent. Key genes involved in Cu metabolism were sequenced for the first time in P. vivipara. Transcripts coding for the high-affinity copper transporter (CTR1) and copper-transporting ATPase (ATP7B) were identified using polymerase chain reaction (PCR) and gene sequencing. The full-length CTR1 open reading frame (1560bp) and a partial ATP7B (690bp) were discovered. Predicted amino acid sequences shared high identities with the CTR1 of Fundulus heteroclitus (81%) and the ATP7B of Sparus aurata (87%). Basal transcriptional levels addressed by RT-qPCR in control fish indicate that CTR1 and ATP7B was highly transcribed in liver of freshwater guppies while CTR1 was highly transcribed in gut of saltwater guppies. This could explain the higher Cu accumulation observed in liver of freshwater guppies and in gut of saltwater guppies, because CTR1 is involved in Cu uptake. Reduced gill mRNA expression of CTR1 was observed in freshwater guppies exposed to 20.0μg/L Cu and in saltwater guppies exposed to 5.0μg/L Cu. In turn, reduced mRNA expression of gut ATP7B was observed in freshwater and salt water guppies exposed to 9.0 and 20.0μg/L Cu. Liver CTR1 and ATP7B transcription were not affected by Cu exposure. These findings suggest that gill CTR1 and gut ATP7B are down-regulated to limit Cu absorption after exposure to dissolved Cu, while liver CTR1 and ATP7B levels are maintained to allow Cu storage and detoxification. In conclusion, findings reported here

  15. Genetic variability in copper-transporting P-type adenosine triphosphatase (ATP7B) is associated with Alzheimer's disease in a Chinese population.

    PubMed

    Liu, H P; Lin, W Y; Wang, W F; Tsai, C H; Wu, W C; Chiou, M T; Shen, C P; Wu, B T; Tsai, F J

    2013-01-01

    Previous experiments demonstrated that transgenic mice carrying both amyloid precursor protein and mutant ATP7B transgenes reduce amyloid plaques and diminish plasma Abeta levels. These experiments showed that a structural change of ATP7B may affect Alzheimer’s disease (AD) susceptibility. In this study three missense SNPs in ATP7B gene (rs1801243, rs1801244, and rs1801249) were chosen to test whether they were associated with AD. We tested this hypothesis using a case control design. The experimental data showed that there was a significant deviation from Hardy-Weinberg equilibrium (HWE) for SNP rs1801249 (c.3419 T greater than C, Val1140Ala) in the case group (p = 0.014) but not in the control group and that there was an association between SNP rs1801249 and AD under a recessive model (p = 0.003). The data also showed that the genotype frequency distribution of the ATP7B c.1366 G greater than C polymorphism (rs1801244, Val456Leu) differed significantly between the AD patients and the normal subjects (p = 0.012). In addition, the frequency of the TGC haplotype of SNPs rs1801243, rs1801244, and rs1801249 was significantly higher in the AD patients compared with the normal subjects (p = 8.49×10-7). These observations suggested that genetic variations in the copper transporter gene ATP7B might contribute to AD pathogenesis in the Taiwanese population.

  16. Early gestational gene transfer with targeted ATP7B expression in the liver improves phenotype in a murine model of Wilson's disease.

    PubMed

    Roybal, J L; Endo, M; Radu, A; Gray, L; Todorow, C A; Zoltick, P W; Lutsenko, S; Flake, A W

    2012-11-01

    The ideal gene therapy for metabolical liver disorders would target hepatocytes before the onset of disease and be durable, non-toxic and non-immunogenic. Early gestational gene transfer can achieve such goals. Here, we demonstrate that prenatal gene transfer of human Atp7b reduces liver pathology and improves biochemical markers in Atp7b(-/-) mice, a murine model of Wilson's disease (WD). Following prenatal injection of lentivirus vector containing the human Atp7b gene under the transcriptional control of a liver-specific promoter, the full-length ATP7B was detectable in mouse livers for the entire duration of experiments (20 weeks after birth). In contrast to a marked pathology in non-injected animals, livers from age-matched treated mice consistently demonstrated normal gross and histological morphology. Hepatic copper content was decreased in the majority of treated mice, although remaining copper levels varied. Improvement of hepatic copper metabolism was further apparent from the presence of copper-bound ceruloplasmin in the sera and normalization of the mRNA levels for HMG CoA-reductase. With this approach, the complete loss of copper transport function can be ameliorated, as evident from phenotypical improvement in treated Atp7b(-/-) mice. This study provides proof of principle for in utero gene therapy in WD and other liver-based enzyme deficiencies.

  17. Alpaca fiber growth is mediated by microRNA let-7b via down-regulation of target gene FGF5.

    PubMed

    Wang, T; Zhang, Y; Wang, H D; Shen, Y; Liu, N; Cao, J; Yu, X J; Dong, C S; He, X Y

    2015-10-29

    MicroRNAs are very small endogenous RNA molecules that play a crucial role in an array of biological processes, including regulation of skin morphogenesis. The microRNA let-7b is thought to modulate animal hair growth, by binding target genes that encode growth factors. Fibroblast growth factor 5 (FGF5) has been previously reported to be involved in the initiation of the catagen phase of hair growth. In this study, we combined previous reports with bioinformatic analysis techniques to identify and validate FGF5 and, using lucerifase assay, confirmed targeted binding of let-7b to FGF5. To investigate the interaction between let-7b and FGF5, alpaca skin fibroblasts were transfected with let-7b over-expression vectors, and then mRNA and protein expression levels of FGF5 and the gene encoding its receptor, FGFR1, were evaluated. Levels of FGF5 mRNA and protein were remarkably lower in transfected groups, as compared to controls. In summary, this study confirmed that let-7b acts as a regulator of skin morphogenesis, by directly targeting FGF5 and down-regulating its expression. It provides the evidence of hair growth regulated by miRNAs in animals and may have important applications in wool production.

  18. KEPLER'S OPTICAL SECONDARY ECLIPSE OF HAT-P-7b AND PROBABLE DETECTION OF PLANET-INDUCED STELLAR GRAVITY DARKENING

    SciTech Connect

    Morris, Brett M.; Deming, Drake; Mandell, Avi M.

    2013-02-20

    We present observations spanning 355 orbital phases of HAT-P-7 observed by Kepler from 2009 May to 2011 March (Q1-9). We find a shallower secondary eclipse depth than initially announced, consistent with a low optical albedo and detection of nearly exclusively thermal emission, without a reflected light component. We find an approximately 10 ppm perturbation to the average transit light curve near phase -0.02 that we attribute to a temperature decrease on the surface of the star, phased to the orbit of the planet. This cooler spot is consistent with planet-induced gravity darkening, slightly lagging the sub-planet position due to the finite response time of the stellar atmosphere. The brightness temperature of HAT-P-7b in the Kepler bandpass is T{sub B} = 2733 {+-} 21 K and the amplitude of the deviation in stellar surface temperature due to gravity darkening is approximately -0.18 K. The detection of the spot is not statistically unequivocal due its small amplitude, though additional Kepler observations should be able to verify the astrophysical nature of the anomaly.

  19. Estrogen-mediated regulation of steroid metabolism in rat glial cells; effects on neurosteroid levels via regulation of CYP7B1-mediated catalysis.

    PubMed

    Wicher, Grzegorz; Norlin, Maria

    2015-01-01

    Many neuroactive steroids, including dehydroepiandrosterone (DHEA), pregnenolone, 27-hydroxycholesterol and 17β-estradiol, are known to affect development and function of the brain and nervous system. These and other steroids can undergo tissue and/or cell-specific enzymatic conversions into steroid metabolites. Carefully regulated production of steroids with various physiological effects is important for cells of the nervous system. Astrocytes express many steroidogenic enzymes and are considered important producers of brain steroids. The quantitative roles of different pathways for steroid metabolism in rat astrocytes are not clear. In the current study we examined effects of estrogens on steroid metabolism catalyzed by CYP7B1 and other enzymes in primary cultures of rat astrocytes. The CYP7B1 enzyme, which has been linked to neurodegenerative disease, is involved in the metabolism of several important neurosteroids. In the present study, we found that 7α-hydroxylation, performed by CYP7B1, is the quantitatively most important pathway for DHEA metabolism in rat astrocytes. In addition, our present experiments on catalytic steroid conversions revealed that estrogens significantly suppress the CYP7B1-catalyzed metabolism of not only DHEA but also of pregnenolone and 27-hydroxycholesterol in rat astrocytes. These novel findings point to a regulatory mechanism for control of the cellular levels of these neurosteroids via CYP7B1. Our hypothesis that estrogens can regulate neurosteroid levels via this enzymatic reaction was supported by experiments using ELISA to assay levels of DHEA and pregnenolone in the presence or absence of estrogen. Furthermore, the present results show that estrogen suppresses CYP7B1-catalyzed 7α-hydroxylation also in primary cultures of rat Schwann cells, indicating that regulation by estrogen via this enzyme may be of relevance in both the CNS and the PNS.

  20. The impact of active site protonation on substrate ring conformation in Melanocarpus albomyces cellobiohydrolase Cel7B.

    PubMed

    Schutt, Timothy C; Bharadwaj, Vivek S; Granum, David M; Maupin, C Mark

    2015-07-14

    The ability to utilize biomass as a feedstock for liquid fuel and value-added chemicals is dependent on the efficient and economic utilization of lignin, hemicellulose, and cellulose. In current bioreactors, cellulases are used to convert crystalline and amorphous cellulose to smaller oligomers and eventually glucose by means of cellulase enzymes. A critical component of the enzyme catalyzed hydrolysis reaction is the degree to which the enzyme can facilitate substrate ring deformation from the chair to a more catalytically active conformation (e.g. skewed boat) at the -1 subsite. Presented here is an evaluation of the impact of the protonation state for critical active site residues (i.e. Glu212, Asp214, Glu217, and His228) in Melanocarpus albomyces (Ma) Cellobiohydrolase Cel7B on the substrate's orientation and ring conformation. It is found that the protonation state of the active site can disrupt the intra-enzyme hydrogen bonding network and enhance the sampling of various ring puckering conformations for the substrate ring at the +1 and -1 subsites. In particular it is observed that the protonation state of Asp214 dictates the accessibility of the glycosidic bond to the catalytic acid/base Glu217 by influencing the φ/ψ dihedral angles and the puckering of the ring structure. The protonation-orientation-conformation analysis has revealed an active site that primarily utilizes two highly coupled protonation schemes; one protonation scheme to orient the substrate and generate catalytically favorable substrate geometries and ring puckering conformations and another protonation scheme to hydrolyze the glycosidic bond. In addition to identifying how enzymes utilize protonation state to manipulate substrate geometry, this study identifies possible directions for improving catalytic activity through protein engineering. PMID:26061383

  1. Cell attachment on poly(3-hydroxybutyrate)-poly(ethylene glycol) copolymer produced by Azotobacter chroococcum 7B

    PubMed Central

    2013-01-01

    Background The improvement of biomedical properties, e.g. biocompatibility, of poly(3-hydroxyalkanoates) (PHAs) by copolymerization is a promising trend in bioengineering. We used strain Azotobacter chroococcum 7B, an effective producer of PHAs, for biosynthesis of not only poly(3-hydroxybutyrate) (PHB) and its main copolymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHB-HV), but also alternative copolymer, poly(3-hydroxybutyrate)-poly(ethylene glycol) (PHB-PEG). Results In biosynthesis we used sucrose as the primary carbon source and valeric acid or poly(ethylene glycol) 300 (PEG 300) as additional carbon sources. The chemical structure of PHB-PEG and PHB-HV was confirmed by 1H nuclear-magnetic resonance (1H NMR) analysis. The physico-chemical properties (molecular weight, crystallinity, hydrophilicity, surface energy) and surface morphology of films from PHB copolymers were studied. To study copolymers biocompatibility in vitro the protein adsorption and COS-1 fibroblasts growth on biopolymer films by XTT assay were analyzed. Both copolymers had changed physico-chemical properties compared to PHB homopolymer: PHB-HV and PHB-PEG had less crystallinity than PHB; PHB-HV was more hydrophobic than PHB in contrast to PHB-PEG appeared to have greater hydrophilicity than PHB; whereas the morphology of polymer films did not differ significantly. The protein adsorption to PHB-PEG was greater and more uniform than to PHB and PHB-PEG copolymer promoted better growth of COS-1 fibroblasts compared with PHB homopolymer. Conclusions Thus, despite low EG-monomers content in bacterial origin PHB-PEG copolymer, this polymer demonstrated significant improvement in biocompatibility in contrast to PHB and PHB-HV copolymers, which may be coupled with increased protein adsorption and hydrophilicity of PEG-containing copolymer. PMID:23692611

  2. Presence of Not5 and ubiquitinated Rps7A in polysome fractions depends upon the Not4 E3 ligase.

    PubMed

    Panasenko, Olesya O; Collart, Martine A

    2012-02-01

    In this study, we determine that Saccharomyces cerevisiae Not4 E3 ligase ubiquitinates Rps7A in vivo and in vitro, but not its paralogue, Rps7B. Ubiquitinated Rps7A is detectable only in 80S and polysomes, but not in free 40S fractions. A different role of the Rps7 paralogues in vivo is supported by the observation that the deletion of Rps7A but not Rps7B is sensitive to translational inhibitors and leads to an accumulation of aggregated proteins. An important accumulation of aggregated proteins that include ribosomal proteins and ribosome-associated chaperones is also observed in cells lacking Not4. A contribution of Not4 to ribosomal function extending beyond Rps7A ubiquitination is supported by the observation that the deletion of Not4 displays a synthetic slow growth phenotype when combined with the deletion of either one of the two Rps7 paralogues. Not4 is detectable in polysome fractions, as are other subunits of the Ccr4-Not complex such as Not5. The optimal presence of Not5 in polysomes is dependent upon Not4 and the deletion of Not5 leads to a dramatic reduction of polysomes. These results lead us to suggest that Not4 contributes to normal polysome levels and is important for cellular protein solubility maybe in part by ubiquitination of Rps7A.

  3. Hidden carbon in Earth’s inner core revealed by shear softening in dense Fe7C3

    PubMed Central

    Li, Zeyu; Zhang, Dongzhou; Liu, Jiachao; Hu, Michael Y.; Zhao, Jiyong; Bi, Wenli; Alp, E. Ercan; Xiao, Yuming; Chow, Paul; Li, Jie

    2014-01-01

    Earth’s inner core is known to consist of crystalline iron alloyed with a small amount of nickel and lighter elements, but the shear wave (S wave) travels through the inner core at about half the speed expected for most iron-rich alloys under relevant pressures. The anomalously low S-wave velocity (vS) has been attributed to the presence of liquid, hence questioning the solidity of the inner core. Here we report new experimental data up to core pressures on iron carbide Fe7C3, a candidate component of the inner core, showing that its sound velocities dropped significantly near the end of a pressure-induced spin-pairing transition, which took place gradually between 10 GPa and 53 GPa. Following the transition, the sound velocities increased with density at an exceptionally low rate. Extrapolating the data to the inner core pressure and accounting for the temperature effect, we found that low-spin Fe7C3 can reproduce the observed vS of the inner core, thus eliminating the need to invoke partial melting or a postulated large temperature effect. The model of a carbon-rich inner core may be consistent with existing constraints on the Earth's carbon budget and would imply that as much as two thirds of the planet's carbon is hidden in its center sphere. PMID:25453077

  4. TTC7B emerges as a novel risk factor for ischemic stroke through the convergence of several genome-wide approaches

    PubMed Central

    Krug, Tiago; Gabriel, João Paulo; Taipa, Ricardo; Fonseca, Benedita V; Domingues-Montanari, Sophie; Fernandez-Cadenas, Israel; Manso, Helena; Gouveia, Liliana O; Sobral, João; Albergaria, Isabel; Gaspar, Gisela; Jiménez-Conde, Jordi; Rabionet, Raquel; Ferro, José M; Montaner, Joan; Vicente, Astrid M; Silva, Mário Rui; Matos, Ilda; Lopes, Gabriela; Oliveira, Sofia A

    2012-01-01

    We hereby propose a novel approach to the identification of ischemic stroke (IS) susceptibility genes that involves converging data from several unbiased genetic and genomic tools. We tested the association between IS and genes differentially expressed between cases and controls, then determined which data mapped to previously reported linkage peaks and were nominally associated with stroke in published genome-wide association studies. We first performed gene expression profiling in peripheral blood mononuclear cells of 20 IS cases and 20 controls. Sixteen differentially expressed genes mapped to reported whole-genome linkage peaks, including the TTC7B gene, which has been associated with major cardiovascular disease. At the TTC7B locus, 46 tagging polymorphisms were tested for association in 565 Portuguese IS cases and 520 controls. Markers nominally associated in at least one test and defining associated haplotypes were then examined in 570 IS Spanish cases and 390 controls. Several polymorphisms and haplotypes in the intron 5–intron 6 region of TTC7B were also associated with IS risk in the Spanish and combined data sets. Multiple independent lines of evidence therefore support the role of TTC7B in stroke susceptibility, but further work is warranted to identify the exact risk variant and its pathogenic potential. PMID:22453632

  5. PROCEEDINGS: 1991 INTERNATIONAL CONFERENCE ON MUNICIPAL WASTE COMBUSTION - VOLUME 2. SESSIONS 1B, 2B, 3B, 4B, 7A, 7B, 8A, 8B, AND 9B

    EPA Science Inventory

    The three-volumes document 82 presentations by authors from 15 countries at the Second International Conference on Municipal Waste Combustion (MWC) in Tampa, Florida, April 16-19, 1991. The Conference fostered the exchange of current information on research concerning MWC, ash di...

  6. Spectrum of mutations in the ATP binding domain of ATP7B gene of Wilson Disease in a regional Indian cohort.

    PubMed

    Guggilla, Sreenivasa Rao; Senagari, Jalandhar Reddy; Rao, P N; Madireddi, Sujatha

    2015-09-10

    Wilson disease is an autosomal recessive disorder of abnormal copper accumulation in the liver, brain, kidney and cornea, resulting in hepatic and neurological abnormalities, which results from impaired ATP7B protein function due to mutations in candidate ATP7B gene, till date more than 500 disease causing mutations were found. In India most disease causing mutations were identified in ATP-BD. DNA samples of the 101 WD cases and 100 control population were analyzed for mutations. 11 mutations were identified in 57 chromosomes. Three novel mutations, c.3310T>A (p.Cys1104Ser), c.3337C>A (p.Leu1113Met) on exon 15 and c.3877G>A (p.Glu1293Lys) on exon 18 were identified for the first time in the ATP7B gene. Two mutations, c.3121C>T (p.Arg1041Trp) and c.3128T>C (p.Leu1043Pro) on exon 14 were discovered for the first time in Indian Wilson disease patients. Four previously reported mutations c.3008C>T, c.3029A>G on exon 13, c.3182G>A on exon 14 and c.3809A>G on exon 18 from South India were also found in this study. Our research has enriched the spectrum of mutations of the ATP7B gene in the south Indian population. The detection of new mutations in the ATP7B gene can aid in genetic counseling and clinical or/prenatal diagnosis.

  7. Downregulation of HMGA2 by the pan-deacetylase inhibitor panobinostat is dependent on hsa-let-7b expression in liver cancer cell lines

    SciTech Connect

    Di Fazio, Pietro; Montalbano, Roberta; Neureiter, Daniel; Alinger, Beate; Schmidt, Ansgar; Merkel, Anna Lena; Quint, Karl; Ocker, Matthias

    2012-09-10

    Inhibitors of protein deacetylases represent a novel therapeutic option for cancer diseases due to their effects on transcriptional regulation by interfering with histones acetylation and on several other cellular pathways. Recently, their ability to modulate several transcription factors and, interestingly, also co-factors, which actively participate in formation and modulation of transcription complexes was shown. We here investigate whether HMGA2 (High Mobility Group AT-2 hook), a nuclear non-histone transcriptional co-factor with known oncogenic properties, can be influenced by the novel pan-deacetylase inhibitor panobinostat (LBH589) in human hepatocellular carcinoma models. Panobinostat strongly downregulated HMGA2 in HepG2 and Hep3B cells; this effect was mediated by transcriptional upregulation and promotion of the maturation of the tumorsuppressor miRNA hsa-let-7b, which could inhibit HMGA2 expression via RNA interference pathways. siRNA knockdown of HMGA2 or transfection of hsa-let-7b mimicking oligonucleotides confirmed the role of HMGA2 in regulating cell proliferation and apoptosis in liver cancer cell lines. Co-incubation with panobinostat showed an additive effect on inhibition of cell proliferation using an impedance-based real-time cell analyzer. Treatment of HepG2 xenografts with panobinostat also led to a downregulation of HMGA2 in vivo. These findings show that pan-deacetylase inhibitors also modulate other signaling pathways and networks than histone modifications to influence cell fate. -- Highlights: Black-Right-Pointing-Pointer Panobinostat for the treatment of liver cancer. Black-Right-Pointing-Pointer Panobinostat meddles with miRNAs-dependent transcriptional and translational control. Black-Right-Pointing-Pointer Tumorsuppressor miRNA hsa-let-7b upregulation. Black-Right-Pointing-Pointer HMGA2 is downregulated via RNA interference pathways mediated by hsa-let-7b. Black-Right-Pointing-Pointer Panobinostat determines inhibition of

  8. Economic assessments of small-scale drinking-water interventions in pursuit of MDG target 7C.

    PubMed

    Cameron, John; Jagals, Paul; Hunter, Paul R; Pedley, Steve; Pond, Katherine

    2011-12-01

    This paper uses an applied rural case study of a safer water intervention in South Africa to illustrate how three levels of economic assessment can be used to understand the impact of the intervention on people's well-being. It is set in the context of Millennium Development Goal 7 which sets a target (7C) for safe drinking-water provision and the challenges of reaching people in remote rural areas with relatively small-scale schemes. The assessment moves from cost efficiency to cost effectiveness to a full social cost-benefit analysis (SCBA) with an associated sensitivity test. In addition to demonstrating techniques of analysis, the paper brings out many of the challenges in understanding how safer drinking-water impacts on people's livelihoods. The SCBA shows the case study intervention is justified economically, though the sensitivity test suggests 'downside' vulnerability.

  9. Microstructure and Dry Sliding Wear Behavior of Fe-Based (Cr, Fe)7C3 Composite Coating Fabricated by PTA Welding Process

    NASA Astrophysics Data System (ADS)

    Yuan, Y. L.; Li, Z. G.

    2013-11-01

    Using Cr3C2 and Fe-CrNiBSi powder blends as raw materials, an α-Fe matrix composite coating reinforced by in situ (Cr, Fe)7C3 rods, with a thickness of about 3.6 mm, was fabricated on the surface of AISI A36 low carbon steel by means of plasma-transferred arc welding. The results of microstructural analysis show that in the coating, a large number of carbides, (Cr, Fe)7C3, in rod shape grow, and radiate around some half-dissolved Cr3C2 particles. The results of dry sliding wear tests at loads 100, 200, and 300 N show that the wear resistances of (Cr, Fe)7C3-reinforced coating, respectively, are about 6.9, 14.9, and 17 times higher than that of nonreinforced pure Fe-CrNiBSi alloy coating; the average value and fluctuation range of friction coefficient (FC) of (Cr, Fe)7C3-reinforced coating are less than those of pure Fe-CrNiBSi alloy coating; the main wear mechanisms of pure Fe-CrNiBSi alloy coating are ploughing, deformation, and adhesive wear, whereas those of (Cr, Fe)7C3-reinforced coating are microcutting, abrasive, and oxidation wear; the cracks on surfaces of (Cr, Fe)7C3 rods increased with the increasing loads; and the matrix α-Fe can prevent them from extending further in the composite coating.

  10. EOS7C Version 1.0: TOUGH2 Module for Carbon Dioxide or Nitrogen inNatural Gas (Methane) Reservoirs

    SciTech Connect

    Oldenburg, Curtis M.; Moridis,George J.; Spycher, Nicholas; Pruess, Karsten

    2004-06-29

    EOS7C is a TOUGH2 module for multicomponent gas mixtures in the systems methane carbon dioxide (CH4-CO2) or methane-nitrogen (CH4-N2) with or without an aqueous phase and H2O vapor. EOS7C uses a cubic equation of state and an accurate solubility formulation along with a multiphase Darcy s Law to model flow and transport of gas and aqueous phase mixtures over a wide range of pressures and temperatures appropriate to subsurface geologic carbon sequestration sites and natural gas reservoirs. EOS7C models supercritical CO2 and subcritical CO2 as a non-condensible gas, hence EOS7C does not model the transition to liquid or solid CO2 conditions. The components modeled in EOS7C are water, brine, non-condensible gas, gas tracer, methane, and optional heat. The non-condensible gas (NCG) can be selected by the user to be CO2 or N2. The real gas properties module has options for Peng-Robinson, Redlich-Kwong, or Soave-Redlich-Kwong equations of state to calculate gas mixture density, enthalpy departure, and viscosity. Partitioning of the NCG and CH4 between the aqueous and gas phases is calculated using a very accurate chemical equilibrium approach. Transport of the gaseous and dissolved components is by advection and Fickian molecular diffusion. We present instructions for use and example problems to demonstrate the accuracy and practical application of EOS7C.

  11. LiVP2O7/C: A New Insertion Anode Material for High-Rate Lithium-Ion Battery Applications.

    PubMed

    Mani, Vellaisamy; Kalaiselvi, Nallathamby

    2016-04-18

    LiVP2O7/C, popularly known so far as an environmentally compatible and economically viable lithium battery cathode material, was exploited for the first time as an anode through the current study. LiVP2O7/C was synthesized by adopting oxalyl dihydrazide assisted solution combustion method and explored as an anode material in rechargeable lithium cell assembly. Notably, an initial capacity of 600 mAh g(-1) was exhibited by LiVP2O7/C anode, at the rate of 0.5 C along with an excellent Coulombic efficiency of 99% up to 150 cycles. The title anode demonstrates its suitability for high capacity and high rate applications by way of exhibiting appreciable capacity values of 200, 150, 120, and 110 mAh g(-1), under the influence of 2, 4, 6, and 8 C rates, respectively. Further, LiVP2O7/C anode, when subjected to a high current 10 C rate, exhibits an acceptable capacity of 107 mAh g(-1) up to 500 cycles, which is closer to its theoretical capacity value of 117 mAh g(-1). The study demonstrates the possibility of exploiting LiVP2O7/C as yet another potential anode and thereby opens a newer avenue to explore wide variety of LiMP2O7/C composites for their probable anode behavior in rechargeable lithium batteries.

  12. Quantitative Measurement of the Proportions of High-Order Harmonics for the 4B7B Soft-X-Ray Source at Beijing Synchrotron Radiation Facility

    NASA Astrophysics Data System (ADS)

    Zhu, Tuo; Shang, Wanli; Zhang, Wenhai; Yang, Jiamin; Xiong, Gang; Zhao, Yang; Kuang, Longyu; Zhao, Yidong; Zheng, Lei; Cui, Mingqi; Tang, Kun; Ma, Chenyan

    2013-12-01

    A transmission grating coupled with an X-ray charge coupled device (CCD) is used to quantitatively measure the proportion of high-order harmonics of the soft-X-ray source of beam line 4B7B. The results show that the monochromatic X-ray has third-order and second-order harmonics. The proportion of second-order harmonic of 4B7B is less than 9.0% and the third-order harmonic is below 0.7% when no suppressing method is applied. When suppression methods are used, the proportion of second-order harmonic is less than 1.7% and the third-order harmonic is ignorable.

  13. Identification of p38 MAPK and JNK as new targets for correction of Wilson disease‐causing ATP7B mutants

    PubMed Central

    Chesi, Giancarlo; Hegde, Ramanath N.; Iacobacci, Simona; Concilli, Mafalda; Parashuraman, Seetharaman; Festa, Beatrice Paola; Polishchuk, Elena V.; Di Tullio, Giuseppe; Carissimo, Annamaria; Montefusco, Sandro; Canetti, Diana; Monti, Maria; Amoresano, Angela; Pucci, Piero; van de Sluis, Bart; Lutsenko, Svetlana

    2016-01-01

    Wilson disease (WD) is an autosomal recessive disorder that is caused by the toxic accumulation of copper (Cu) in the liver. The ATP7B gene, which is mutated in WD, encodes a multitransmembrane domain adenosine triphosphatase that traffics from the trans‐Golgi network to the canalicular area of hepatocytes, where it facilitates excretion of excess Cu into the bile. Several ATP7B mutations, including H1069Q and R778L that are two of the most frequent variants, result in protein products, which, although still functional, remain in the endoplasmic reticulum. Thus, they fail to reach Cu excretion sites, resulting in the toxic buildup of Cu in the liver of WD patients. Therefore, correcting the location of these mutants by leading them to the appropriate functional sites in the cell should restore Cu excretion and would be beneficial to help large cohorts of WD patients. However, molecular targets for correction of endoplasmic reticulum‐retained ATP7B mutants remain elusive. Here, we show that expression of the most frequent ATP7B mutant, H1069Q, activates p38 and c‐Jun N‐terminal kinase signaling pathways, which favor the rapid degradation of the mutant. Suppression of these pathways with RNA interference or specific chemical inhibitors results in the substantial rescue of ATP7BH1069Q (as well as that of several other WD‐causing mutants) from the endoplasmic reticulum to the trans‐Golgi network compartment, in recovery of its Cu‐dependent trafficking, and in reduction of intracellular Cu levels. Conclusion: Our findings indicate p38 and c‐Jun N‐terminal kinase as intriguing targets for correction of WD‐causing mutants and, hence, as potential candidates, which could be evaluated for the development of novel therapeutic strategies to combat WD. (Hepatology 2016;63:1842‐1859) PMID:26660341

  14. Expression of WNT5A in Idiopathic Pulmonary Fibrosis and Its Control by TGF-β and WNT7B in Human Lung Fibroblasts.

    PubMed

    Newman, Donna R; Sills, W Shane; Hanrahan, Katherine; Ziegler, Amanda; Tidd, Kathleen McGinnis; Cook, Elizabeth; Sannes, Philip L

    2016-02-01

    The wingless (Wnt) family of signaling ligands contributes significantly to lung development and is highly expressed in patients with usual interstitial pneumonia (UIP). We sought to define the cellular distribution of Wnt5A in the lung tissue of patients with idiopathic pulmonary fibrosis (IPF) and the signaling ligands that control its expression in human lung fibroblasts and IPF myofibroblasts. Tissue sections from 40 patients diagnosed with IPF or UIP were probed for the immunolocalization of Wnt5A. Further, isolated lung fibroblasts from normal or IPF human lungs, adenovirally transduced for the overexpression or silencing of Wnt7B or treated with TGF-β1 or its inhibitor, were analyzed for Wnt5A protein expression. Wnt5A was expressed in IPF lungs by airway and alveolar epithelium, smooth muscle cells, endothelium, and myofibroblasts of fibroblastic foci and throughout the interstitium. Forced overexpression of Wnt7B with or without TGF-β1 treatment significantly increased Wnt5A protein expression in normal human smooth muscle cells and fibroblasts but not in IPF myofibroblasts where Wnt5A was already highly expressed. The results demonstrate a wide distribution of Wnt5A expression in cells of the IPF lung and reveal that it is significantly increased by Wnt7B and TGF-β1, which, in combination, could represent key signaling pathways that modulate the pathogenesis of IPF.

  15. IGF-1R, a target of let-7b, mediates crosstalk between IRS-2/Akt and MAPK pathways to promote proliferation of oral squamous cell carcinoma

    PubMed Central

    Gao, Ling; Wang, Xiaolong; Wang, Xiaofei; Zhang, Linmei; Qiang, Cui; Chang, Su'e; Ren, Wenhao; Li, Shaoming; Yang, Yang; Tong, Dongdong; Chen, Cheng; Li, Zongfang; Song, Tusheng; Zhi, Keqian; Huang, Chen

    2014-01-01

    Insulin-like growth factor (IGF) signaling is involved in oral squamous cell carcinoma (OSCC), but IGF-1 receptor (IGF-1R)-mediated intricate regulatory networks among molecular interactions and signalling path ways in OSCC remain unclear. Here, we found that overexpression of IGF-1R and insulin receptor substrate-2 (IRS-2) was negatively associated with histological differentiation. IGF signaling stimulated OSCC cell growth. Conversely, overexpression of let-7b inhibited proliferation and colony formation and triggered S/G2 cell cycle arrest by targeting IGF-1R and IRS-2 through the Akt pathway. Also, the inverse relationship between expression of let-7b and IGF-1R/IRS-2 was confirmed in OSCC tumor xenografts and clinical specimens. Furthermore, by activating ERK1/2, IGF-1R transcriptionally upregulated IRS-2. Our results indicate that let-7b/IGF-1R-mediated crosstalk between IRS-2/Akt and MAPK is involved in OSCC and is a potential therapeutic target for therapy. PMID:24810113

  16. Abnormal epigenetic regulation of the gene expression levels of Wnt2b and Wnt7b: Implications for neural tube defects

    PubMed Central

    BAI, BAOLING; CHEN, SHUYUAN; ZHANG, QIN; JIANG, QIAN; LI, HUILI

    2016-01-01

    The association between Wnt genes and neural tube defects (NTDs) is recognized, however, it remains to be fully elucidated. Our previous study demonstrated that epigenetic mechanisms are affected in human NTDs. Therefore, the present study aimed to evaluate whether Wnt2b and Wnt7b are susceptible to abnormal epigenetic modification in NTDs, using chromatin immunoprecipitation assays to evaluate histone enrichments and the MassARRAY platform to detect the methylation levels of target regions within Wnt genes. The results demonstrated that the transcriptional activities of Wnt2b and Wnt7b were abnormally upregulated in mouse fetuses with NTDs and, in the GC-rich promoters of these genes, histone 3 lysine 4 (H3K4) acetylation was enriched, whereas H3K27 trimethylation was reduced. Furthermore, several CpG sites in the altered histone modification of target regions were significantly hypomethylated. The present study also detected abnormal epigenetic modifications of these Wnt genes in human NTDs. In conclusion, the present study detected abnormal upregulation in the levels of Wnt2b and Wnt7b, and hypothesized that the alterations may be due to the ectopic opening of chromatin structure. These results improve understanding of the dysregulation of epigenetic modification of Wnt genes in NTDs. PMID:26548512

  17. 3.6 and 4.5 μm Spitzer Phase Curves of the Highly Irradiated Hot Jupiters WASP-19b and HAT-P-7b

    NASA Astrophysics Data System (ADS)

    Wong, Ian; Knutson, Heather A.; Kataria, Tiffany; Lewis, Nikole K.; Burrows, Adam; Fortney, Jonathan J.; Schwartz, Joel; Shporer, Avi; Agol, Eric; Cowan, Nicolas B.; Deming, Drake; Désert, Jean-Michel; Fulton, Benjamin J.; Howard, Andrew W.; Langton, Jonathan; Laughlin, Gregory; Showman, Adam P.; Todorov, Kamen

    2016-06-01

    We analyze full-orbit phase curve observations of the transiting hot Jupiters WASP-19b and HAT-P-7b at 3.6 and 4.5 μm, obtained using the Spitzer Space Telescope. For WASP-19b, we measure secondary eclipse depths of 0.485%+/- 0.024% and 0.584%+/- 0.029% at 3.6 and 4.5 μm, which are consistent with a single blackbody with effective temperature 2372 ± 60 K. The measured 3.6 and 4.5 μm secondary eclipse depths for HAT-P-7b are 0.156%+/- 0.009% and 0.190%+/- 0.006%, which are well described by a single blackbody with effective temperature 2667 ± 57 K. Comparing the phase curves to the predictions of one-dimensional and three-dimensional atmospheric models, we find that WASP-19b’s dayside emission is consistent with a model atmosphere with no dayside thermal inversion and moderately efficient day-night circulation. We also detect an eastward-shifted hotspot, which suggests the presence of a superrotating equatorial jet. In contrast, HAT-P-7b’s dayside emission suggests a dayside thermal inversion and relatively inefficient day-night circulation; no hotspot shift is detected. For both planets, these same models do not agree with the measured nightside emission. The discrepancies in the model-data comparisons for WASP-19b might be explained by high-altitude silicate clouds on the nightside and/or high atmospheric metallicity, while the very low 3.6 μm nightside planetary brightness for HAT-P-7b may be indicative of an enhanced global C/O ratio. We compute Bond albedos of 0.38 ± 0.06 and 0 (\\lt 0.08 at 1σ ) for WASP-19b and HAT-P-7b, respectively. In the context of other planets with thermal phase curve measurements, we show that WASP-19b and HAT-P-7b fit the general trend of decreasing day-night heat recirculation with increasing irradiation.

  18. Decreased expression of let-7c is associated with non-response of muscle-invasive bladder cancer patients to neoadjuvant chemotherapy.

    PubMed

    Vinall, Ruth L; Tepper, Clifford G; Ripoll, Alexandra A Z; Gandour-Edwards, Regina F; Durbin-Johnson, Blythe P; Yap, Stanley A; Ghosh, Paramita M; deVere White, Ralph W

    2016-03-01

    The identification and development of biomarkers which predict response of muscle invasive bladder cancer (MIBC) patients to neoadjuvant chemotherapy would likely increase usage of this treatment option and thereby improve patient survival rates. MiRNA array and qRT-PCR validation was used to identify miRNA which are associated with response to neoadjuvant chemotherapy. RNA was extracted from a total of 41 archival, fully annotated, MIBC patient diagnostic biopsies (20 chemo-responders and 21 non-responders (response is defined as > 5 year survival rate and being pT0 post-chemotherapy)). Microarray and qPCR identified let-7c as being differentially expressed in chemo-responder versus non-responder patients. Patients with higher let-7c expression levels had significantly higher odds of responding to chemotherapy (p = 0.023, OR 2.493, 95% CI 1.121, 5.546), and assessment of let-7c levels allowed for prediction of patient response (AUC 0.72, positive predictive value 59%). Decreased let-7c was associated with MIBC incidence (p < 0.001), and significantly correlated with other related miRNA including those that were not differentially expressed between responders and non-responders. The combined data indicate let-7c plays a role in mediating chemoresistance to neoadjuvant chemotherapy in MIBC patients, and is a modest, yet clinically meaningful, predictor of patient response. PMID:27382433

  19. The DNA-recognition fold of Sso7c4 suggests a new member of SpoVT-AbrB superfamily from archaea

    PubMed Central

    Hsu, Chun-Hua; Wang, Andrew H.-J.

    2011-01-01

    Organisms growing at elevated temperatures face the challenge of maintaining the integrity of their genetic materials. Archaea possess unique chromatin proteins for gene organization and information processing. We present the solution structure of Sso7c4 from Sulfolobus solfataricus, which has a homodimeric DNA-binding fold forming a swapped β-loop-β ‘Tai-Chi’ topology. The fold is reminiscent of the N-terminal DNA-binding domain of AbrB and MazE. In addition, several amide resonances in the heteronuclear single quantum coherence spectra of Sso7c4 are shifted and broadened with the addition of small amounts of duplex DNA oligomers. The locations of the corresponding amides in the Sso7c4 structure define its DNA-interacting surface. NMR spectra of DNA titrated with the protein further indicated that Sso7c4 interacts with DNA in the major groove. Taken together, a plausible model for the Sso7c4–DNA complex is presented, in which the DNA double helix is curved around the protein dimer. PMID:21546550

  20. Characterization of a purified thermostable xylanase from Caldicoprobacter algeriensis sp. nov. strain TH7C1(T).

    PubMed

    Amel, Bouanane-Darenfed; Nawel, Boucherba; Khelifa, Bouacem; Mohammed, Gagaoua; Manon, Joseph; Salima, Kebbouche-Gana; Farida, Nateche; Hocine, Hacene; Bernard, Ollivier; Jean-Luc, Cayol; Marie-Laure, Fardeau

    2016-01-01

    The present study investigates the purification and biochemical characterization of an extracellular thermostable xylanase (called XYN35) from Caldicoprobacter algeriensis sp. nov., strain TH7C1(T), a thermophilic, anaerobic strain isolated from the hydrothermal hot spring of Guelma (Algeria). The maximum xylanase activity recorded after 24 h of incubation at 70 °C and in an optimized medium containing 10 g/L mix birchwood- and oats spelt-xylan was 250 U/mL. The pure protein was obtained after heat treatment (1 h at 70 °C), followed by sequential column chromatographies on Sephacryl S-200 gel filtration and Mono-S Sepharose anion-exchange. Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis indicated that the purified enzyme is a monomer with a molecular mass of 35,075.10 Da. The results from amino-acid sequence analysis revealed high homology between the 21 NH2-terminal residues of XYN35 and those of bacterial xylanases. The enzyme showed optimum activity at pH 11 and 70 °C. While XYN35 was activated by Ca(2+), Mn(2+), and Mg(2+), it was completely inhibited by Hg(2+) and Cd(2+). The xylanase showed higher specific activity on soluble oat-spelt xylan, followed by beechwood xylan. This enzyme was also noted to obey the Michaelis-Menten kinetics, with Km and kcat values on oat-spelt xylan being 1.33 mg/mL and 400 min(-1), respectively. Thin-layer chromatography soluble oat-spelt xylan (TLC) analysis showed that the final hydrolyzed products of the enzyme from birchwood xylan were xylose, xylobiose, and xylotriose. Taken together, the results indicated that the XYN35 enzyme has a number of attractive biochemical properties that make it a potential promising candidate for future application in the pulp bleaching industry. PMID:26687892

  1. Summary Report For The Analysis Of The Sludge Batch 7b (Macrobatch 9) DWPF Pour Stream Glass Sample For Canister S04023

    SciTech Connect

    Johnson, F. C.

    2013-11-18

    In order to comply with the Defense Waste Processing Facility (DWPF) Waste Form Compliance Plan for Sluldge Batch 7b, Savannah River National Laboratory (SRNL) personnel characterized the Defense Waste Processing Facility (DWPF) pour stream (PS) glass sample collected while filling canister S04023. This report summarizes the results of the compositional analysis for reportable oxides and radionuclides and the normalized Product Consistency Test (PCT) results. The PCT responses indicate that the DWPF produced glass that is significantly more durable than the Environmental Assessment glass.

  2. ORF7-encoded accessory protein 7a of feline infectious peritonitis virus as a counteragent against IFN-α-induced antiviral response.

    PubMed

    Dedeurwaerder, Annelike; Olyslaegers, Dominique A J; Desmarets, Lowiese M B; Roukaerts, Inge D M; Theuns, Sebastiaan; Nauwynck, Hans J

    2014-02-01

    The type I IFN-mediated immune response is the first line of antiviral defence. Coronaviruses, like many other viruses, have evolved mechanisms to evade this innate response, ensuring their survival. Several coronavirus accessory genes play a central role in these pathways, but for feline coronaviruses this has never to our knowledge been studied. As it has been demonstrated previously that ORF7 is essential for efficient replication in vitro and virulence in vivo of feline infectious peritonitis virus (FIPV), the role of this ORF in the evasion of the IFN-α antiviral response was investigated. Deletion of ORF7 from FIPV strain 79-1146 (FIPV-Δ7) rendered the virus more susceptible to IFN-α treatment. Given that ORF7 encodes two proteins, 7a and 7b, it was further explored which of these proteins is active in this mechanism. Providing 7a protein in trans rescued the mutant FIPV-Δ7 from IFN sensitivity, which was not achieved by addition of 7b protein. Nevertheless, addition of protein 7a to FIPV-Δ3Δ7, a FIPV mutant deleted in both ORF3 and ORF7, could no longer increase the replication capacity of this mutant in the presence of IFN. These results indicate that FIPV 7a protein is a type I IFN antagonist and protects the virus from the antiviral state induced by IFN, but it needs the presence of ORF3-encoded proteins to exert its antagonistic function.

  3. The silicate and carbon-rich models of CoRoT-7b, Kepler-9d and Kepler-10b

    NASA Astrophysics Data System (ADS)

    Gong, Yan-Xiang; Zhou, Ji-Lin

    2012-06-01

    Possible bulk compositions of the super-Earth exoplanets CoRoT-7b, Kepler-9d, and Kepler-10b are investigated by applying a commonly used silicate model and a non-standard carbon model. Their internal structures are deduced using a suitable equation of state for the materials. The degeneracy problems of their compositions can be partly overcome, based on the fact that all three planets are extremely close to their host stars. By analyzing the numerical results, we conclude: 1) the iron core of CoRoT-7b is not more than 27% of its total mass within 1σ mass-radius error bars, so an Earth-like composition is less likely, but its carbon rich model can be compatible with an Earth-like core/mantle mass fraction; 2) Kepler-10b is more likely to have a Mercury-like composition, with its old age implying that its high iron content may be a result of strong solar wind or giant impact; 3) the transiting-only super-Earth Kepler-9d is also discussed. Combining its possible composition with the formation theory, we can place some constraints on its mass and bulk composition.

  4. Polycomb-mediated loss of microRNA let-7c determines inflammatory macrophage polarization via PAK1-dependent NF-κB pathway.

    PubMed

    Zhang, W; Liu, H; Liu, W; Liu, Y; Xu, J

    2015-02-01

    Serine/threonine kinase family members p21-activated kinases (PAKs) are important regulators of cytoskeletal remodeling and cell motility in mononuclear phagocytic system, but their role in macrophage differentiation and polarization remains obscure. We have shown here that inflammatory stimuli induced PAK1 overexpression in human and murine macrophages. Elevated expression of PAK1 contributed to macrophage M1 polarization and lipopolysaccharide (LPS)-induced endotoxin shock. We further observed that epigenetic loss of microRNA let-7c due to enhancer of zeste homolog 2 (EZH2) upregulation determined PAK1 elevation and inflammatory phenotype in M1 macrophages. EZH2/let-7c/PAK1 axis promotes macrophage M1 polarization via NIK-IKK-NF-κB signaling. Moreover, pharmacological and genetic ablation with EZH2/let-7c/PAK1 axis blunted inflammatory phenotype in M1 macrophages. Critically, either myeloid-restricted PAK1 deletion (PAK1(Lyz2cre)) or pharmacological and genetic ablation with EZH2/let-7c/PAK1 signal resulted in resistance to LPS-induced endotoxin shock via blunting macrophage M1 polarization. PAK1, therefore, is an essential controller of inflammatory macrophage polarization, regulating immune responses against pathogenic stimuli.

  5. Novel conjugated polymers based on dithieno[3,2-b:6,7-b]carbazole for solution processed thin-film transistors.

    PubMed

    Chen, Yagang; Liu, Chengfang; Tian, Hongkun; Bao, Cheng; Zhang, Xiaojie; Yan, Donghang; Geng, Yanhou; Wang, Fosong

    2012-10-26

    Two conjugated polymers (CPs) P-tCzC12 and P-tCzC16 comprising alternating dithieno[3,2-b:6,7-b]carbazole and 4,4'-dihexadecyl-2,2'-bithiophene units have been designed and synthesized. Upon thermal annealing, they can form ordered thin films in which the polymer backbones dominantly adopted an edge-on orientation respective to the substrate with a lamellar spacing of ≈24 Å and a π-stacking distance of ≈3.7 Å. Organic thin-film transistors (OTFTs) were fabricated by solution casting. A hole mobility of 0.39 cm(2) V(-1) s(-1) has been demonstrated with P-tCzC16. This value is the highest among the CPs containing heteroacenes larger than 4 rings.

  6. Efficient lipid staining in plant material with sudan red 7B or fluorol [correction of fluoral] yellow 088 in polyethylene glycol-glycerol.

    PubMed

    Brundrett, M C; Kendrick, B; Peterson, C A

    1991-01-01

    Polyethylene glycol (400) with 90% glycerol (aqueous) is introduced as an efficient solvent system for lipid stains. Various lipid-soluble dyes were dissolved in this solvent system and tested for their intensity, contrast, and specificity of staining of suberin lamellae in plant tissue. The stability (i.e., lack of precipitation) of the various staining solutions in the presence of fresh tissue was also tested. When dissolved in polyethylene glycol-glycerol, Sudan red 7B (fat red) was the best nonfluorescent stain and fluorol yellow 088 (solvent green 4) was an excellent fluorochrome. These two dyes formed stable staining solutions which efficiently stained lipids in fresh sections without forming precipitates. Estimations of the solubilities of these dyes in the solvent compared with their solubilities in lipids of various chemical types indicated that they should both be effective stains for lipids in general.

  7. Sex-dependent regulation of cytochrome P450 family members Cyp1a1, Cyp2e1, and Cyp7b1 by methylation of DNA

    PubMed Central

    Penaloza, Carlos G.; Estevez, Brian; Han, Dinah M.; Norouzi, Melissa; Lockshin, Richard A.; Zakeri, Zahra

    2014-01-01

    Sexual differences are only partially attributable to hormones. Cultured male or female cells, even from embryos before sexual differentiation, differ in gene expression and sensitivity to toxins, and these differences persist in isolated primary cells. Male and female cells from Swiss Webster CWF mice manifest sex-distinct patterns of DNA methylation for X-ist and for cytochrome P450 (CYP; family members 1a1, 2e1m, and 7b1. Dnmt3l is differentially expressed but not differentially methylated, and Gapdh is neither differentially methylated nor expressed. CYP family genes differ in expression in whole tissue homogenates and cell cultures, with female Cyp expression 2- to 355-fold higher and Dnmt3l 12- to 32-fold higher in males. DNA methylation in the promoters of these genes is sex dimorphic; reducing methylation differences reduces to 1- to 6-fold differences in the expression of these genes. Stress or estradiol alters both methylation and gene expression. We conclude that different methylation patterns partially explain the sex-based differences in expression of CYP family members and X-ist, which potentially leads to inborn differences between males and females and their different responses to chronic and acute changes. Sex-differential methylation may have medical effects.—Penaloza, C.G., Estevez, B., Han, D.M., Norouzi, M., Lockshin, R.A., Zakeri, Z. Sex-dependent regulation of cytochrome P450 family members Cyp1a1, Cyp2e1, and Cyp7b1 by methylation of DNA. PMID:24161885

  8. A SEMI-ANALYTICAL MODEL OF VISIBLE-WAVELENGTH PHASE CURVES OF EXOPLANETS AND APPLICATIONS TO KEPLER- 7 B AND KEPLER- 10 B

    SciTech Connect

    Hu, Renyu; Demory, Brice-Olivier; Seager, Sara; Lewis, Nikole; Showman, Adam P.

    2015-03-20

    Kepler has detected numerous exoplanet transits by measuring stellar light in a single visible-wavelength band. In addition to detection, the precise photometry provides phase curves of exoplanets, which can be used to study the dynamic processes on these planets. However, the interpretation of these observations can be complicated by the fact that visible-wavelength phase curves can represent both thermal emission and scattering from the planets. Here we present a semi-analytical model framework that can be applied to study Kepler and future visible-wavelength phase curve observations of exoplanets. The model efficiently computes reflection and thermal emission components for both rocky and gaseous planets, considering both homogeneous and inhomogeneous surfaces or atmospheres. We analyze the phase curves of the gaseous planet Kepler- 7 b and the rocky planet Kepler- 10 b using the model. In general, we find that a hot exoplanet’s visible-wavelength phase curve having a significant phase offset can usually be explained by two classes of solutions: one class requires a thermal hot spot shifted to one side of the substellar point, and the other class requires reflective clouds concentrated on the same side of the substellar point. Particularly for Kepler- 7 b, reflective clouds located on the west side of the substellar point can best explain its phase curve. The reflectivity of the clear part of the atmosphere should be less than 7% and that of the cloudy part should be greater than 80%, and the cloud boundary should be located at 11° ± 3° to the west of the substellar point. We suggest single-band photometry surveys could yield valuable information on exoplanet atmospheres and surfaces.

  9. Insights into endosomal maturation of human holo-transferrin in the enteric parasite Entamoeba histolytica: essential roles of Rab7A and Rab5 in biogenesis of giant early endocytic vacuoles.

    PubMed

    Verma, Kuldeep; Saito-Nakano, Yumiko; Nozaki, Tomoyoshi; Datta, Sunando

    2015-12-01

    The pathogenic amoeba Entamoeba histolytica is one of the causative agents of health hazards in tropical countries. It causes amoebic dysentery, colitis and liver abscesses in human. Iron is one of the essential nutritional resources for survival and chronic infection caused by the amoeba. The parasite has developed multiple ways to import, sequester and utilize iron from various iron-binding proteins from its host. In spite of its central role in pathogenesis, the mechanism of iron uptake by the parasite is largely unknown. Here, we carried out a systematic study to understand the role of some of the amoebic homologues of mammalian endocytic Rab GTPases (Rab5 and Rab21, Rab7A and Rab7B) in intracellular transport of human holo-transferrin by the parasite. Flow cytometry and quantitative microscopic image analysis revealed that Rab5 and Rab7A are required for the biogenesis of amoebic giant endocytic vacuoles (GEVs) and regulate the early phase of intracellular trafficking of transferrin. Rab7B is involved in the late phase, leading to the degradation of transferrin in the amoebic lysosome-like compartments. Using time-lapse fluorescence imaging in fixed trophozoites, we determined the kinetics of the vesicular transport of transferrin through Rab5-, Rab7A- and Rab7B-positive compartments. The involvement of Rab7A in the early phase of endocytosis by the parasite marks a significant divergence from its host in terms of spatiotemporal regulation by the Rab GTPases. PMID:26096601

  10. Genome-wide screen identified let-7c/miR-99a/miR-125b regulating tumor progression and stem-like properties in cholangiocarcinoma.

    PubMed

    Lin, K-Y; Ye, H; Han, B-W; Wang, W-T; Wei, P-P; He, B; Li, X-J; Chen, Y-Q

    2016-06-30

    Cholangiocarcinoma (CCA), which is a poor prognosis malignancy that arises from the malignant transformation of cholangiocytes, is associated with chronic inflammation of the biliary epithelium. Thus far, the molecular mechanisms of the origin and neoplastic processes of CCA that are promoted by inflammation are still unclear and need to be fully elucidated. Here using small RNA sequencing to determine the microRNA (miRNA) expression profiles in CCA, we found that let-7c, miR-99a and miR-125b, which are three miRNAs of the same cluster, were downregulated in CCA and targeted interleukin 6 (IL-6), IL-6R and type 1 insulin-like growth factor, which are important cytokines and receptors of the IL-6/signal transducer and activator 3 (STAT3) pathway and have key roles in inflammation and CCA initiation. We also found that enforced expression of let-7c, miR-99a or miR-125b could reduce the activity of STAT3 and further suppress CCA tumorigenicity in vivo and inhibit the migration and invasion of CCA cells in vitro. Surprisingly, let-7c/miR-99a/miR-125b cluster also significantly decreased the ability of CCA cells for cancer stem cell-like mammosphere generation by downregulating CD133 and CD44, which suggests the pivotal roles of let-7c, miR-99a and miR-125b in CCA by regulating both inflammation and stem-like properties. Our findings showed potential links between miRNAs and inflammation, and provide a potential treatment strategy for developing an miRNA-based therapy via IL-6/STAT3 targeting for CCA. PMID:26455324

  11. Membrane Docking of the Synaptotagmin 7 C2A Domain: Electron Paramagnetic Resonance Measurements Show Contributions from Two Membrane Binding Loops

    PubMed Central

    Osterberg, J. Ryan; Chon, Nara Lee; Boo, Arthur; Maynard, Favinn A.; Lin, Hai; Knight, Jefferson D.

    2015-01-01

    The synaptotagmin (Syt) family of proteins plays an important role in vesicle docking and fusion during Ca2+-induced exocytosis in a wide variety of cell types. Its role as a Ca2+ sensor derives primarily from its two C2 domains, C2A and C2B, which insert into anionic lipid membranes upon binding Ca2+. Syt isoforms 1 and 7 differ significantly in their Ca2+ sensitivity; the C2A domain from Syt7 binds Ca2+ and membranes much more tightly than the C2A domain from Syt1, due at least in part to greater contributions from the hydrophobic effect. While the structure and membrane activity of Syt1 have been extensively studied, the structural origins of differences between Syt7 and Syt1 are unknown. The present study used site-directed spin labeling and electron paramagnetic resonance spectroscopy to determine depth parameters for the Syt7 C2A domain, for comparison to analogous previous measurements with Syt1 C2A. In a novel approach, the membrane docking geometry of both Syt1 and Syt7 C2A was modeled by mapping depth parameters onto multiple molecular dynamics simulated structures of the Ca2+-bound protein. The models reveal membrane penetration of Ca2+ binding loops (CBLs) 1 and 3, and membrane binding is more sensitive to mutations in CBL3. On average, Syt7 C2A inserts more deeply in the membrane than Syt1 C2A, although depths vary among the different structural models. This observation provides a partial structural explanation for the hydrophobically driven membrane docking of Syt7 C2A. PMID:26322740

  12. Crystal structure and electronic properties of the new compounds, U 6Fe 16Si 7 and its interstitial carbide U 6Fe 16Si 7C

    NASA Astrophysics Data System (ADS)

    Berthebaud, D.; Tougait, O.; Potel, M.; Lopes, E. B.; Gonçalves, A. P.; Noël, H.

    2007-10-01

    The new compounds U6Fe16Si7 and U6Fe16Si7C were prepared by arc-melting and subsequent annealing at 1500 °C. Single-crystal X-ray diffraction showed that they crystallize in the cubic space group Fm3¯m (No. 225), with unit-cell parameters at room temperature a=11.7206(5) Å for U6Fe16Si7 and a=11.7814(2) Å for U6Fe16Si7C. Their crystal structures correspond to ordered variants of the Th6Mn23 type. U6Fe16Si7 adopts the Mg6Cu16Si7 structure type, whereas U6Fe16Si7C crystallizes with a novel "filled" quaternary variant. The inserted carbon is located in octahedral cages formed by six U atoms, with U-U interatomic distances of 3.509(1) Å. Insertion of carbon in the structure of U6Fe16Si7 has a direct influence on the U-Fe and Fe-Fe interatomic distances. The electronic properties of both compounds were investigated by means of DC susceptibility, electrical resistivity and thermopower. U6Fe16Si7 is a Pauli paramagnet. Its electrical resistivity and thermopower point out that it cannot be classified as a simple metal. The magnetic susceptibility of U 6Fe 16Si 7C is best described over the temperature range 100-300 K by using a modified Curie-Weiss law with an effective magnetic moment of 2.3(2) μB/U, a paramagnetic Weiss temperature, θp=57(2) K and a temperature-independent term χ0=0.057(1) emu/mol. Both the electrical resistivity and thermopower reveal metallic behavior.

  13. Magnetic phase diagrams and thermal equations of state of Fe7C3 and Fe3C up to the pressure-temperature conditions of Earth's core

    NASA Astrophysics Data System (ADS)

    Liu, J.; Li, Z.; Chen, B.; Li, J.; Ikuta, D.; Smith, J.; Sinogeikin, S. V.; Popov, D.; Kenney-Benson, C.

    2015-12-01

    Iron carbides Fe7C3 and Fe3C have been proposed as candidates for the dominant components of the Earth's inner core to explain its density deficit and velocity discrepancy with respect to those of pure iron at relevant pressure-temperature conditions (e.g. Chen et al., 2012; Chen et al., 2014; Gao et al., 2011). Testing the hypothesis of carbon-rich inner core requires knowledge of the thermal equations of state of the iron carbides to megabar pressures. Existing experimental data, however, are restricted to either ambient temperature or the pressures near the top of the lower mantle. In particular, the thermal expansion coefficients of the carbides under high pressures remain poorly constrained. Previous studies showed that both Fe7C3 an Fe3C are Invar-type alloys with extremely low thermal expansions in the ferromagnetic phases, and their thermal expansion coefficients more than triple across the ferromagnetic-paramagnetic transition at 1 bar (Litasov et al., 2015). On the other hand, both iron carbides experience pressure-induced spin transitions under high pressures (e.g. Chen et al., 2012; Prescher et al., 2012), but their effects on the thermal expansion are still unknown. We conducted synchrotron X-ray diffraction measurements on Fe7C3 and Fe3C up to the core's pressure and temperature conditions. High pressures up to ~140 GPa were generated by using diamond anvil cells (DAC), while high temperatures were generated using the whole-cell resistive heating device up to 700 K, and using the double-sided laser-heating system up to ~ 3500 K. The new data allow us to construct the magnetic phase diagrams of Fe7C3 and Fe3C and to assess the influences of magnetic transitions on their thermal expansion coefficients. The density profiles of appropriate magnetic phases are calculated and compared with that of the Earth's inner core to estimate its carbon content.

  14. Transiting exoplanets from the CoRoT space mission. VIII. CoRoT-7b: the first super-Earth with measured radius

    NASA Astrophysics Data System (ADS)

    Léger, A.; Rouan, D.; Schneider, J.; Barge, P.; Fridlund, M.; Samuel, B.; Ollivier, M.; Guenther, E.; Deleuil, M.; Deeg, H. J.; Auvergne, M.; Alonso, R.; Aigrain, S.; Alapini, A.; Almenara, J. M.; Baglin, A.; Barbieri, M.; Bruntt, H.; Bordé, P.; Bouchy, F.; Cabrera, J.; Catala, C.; Carone, L.; Carpano, S.; Csizmadia, Sz.; Dvorak, R.; Erikson, A.; Ferraz-Mello, S.; Foing, B.; Fressin, F.; Gandolfi, D.; Gillon, M.; Gondoin, Ph.; Grasset, O.; Guillot, T.; Hatzes, A.; Hébrard, G.; Jorda, L.; Lammer, H.; Llebaria, A.; Loeillet, B.; Mayor, M.; Mazeh, T.; Moutou, C.; Pätzold, M.; Pont, F.; Queloz, D.; Rauer, H.; Renner, S.; Samadi, R.; Shporer, A.; Sotin, Ch.; Tingley, B.; Wuchterl, G.; Adda, M.; Agogu, P.; Appourchaux, T.; Ballans, H.; Baron, P.; Beaufort, T.; Bellenger, R.; Berlin, R.; Bernardi, P.; Blouin, D.; Baudin, F.; Bodin, P.; Boisnard, L.; Boit, L.; Bonneau, F.; Borzeix, S.; Briet, R.; Buey, J.-T.; Butler, B.; Cailleau, D.; Cautain, R.; Chabaud, P.-Y.; Chaintreuil, S.; Chiavassa, F.; Costes, V.; Cuna Parrho, V.; de Oliveira Fialho, F.; Decaudin, M.; Defise, J.-M.; Djalal, S.; Epstein, G.; Exil, G.-E.; Fauré, C.; Fenouillet, T.; Gaboriaud, A.; Gallic, A.; Gamet, P.; Gavalda, P.; Grolleau, E.; Gruneisen, R.; Gueguen, L.; Guis, V.; Guivarc'h, V.; Guterman, P.; Hallouard, D.; Hasiba, J.; Heuripeau, F.; Huntzinger, G.; Hustaix, H.; Imad, C.; Imbert, C.; Johlander, B.; Jouret, M.; Journoud, P.; Karioty, F.; Kerjean, L.; Lafaille, V.; Lafond, L.; Lam-Trong, T.; Landiech, P.; Lapeyrere, V.; Larqué, T.; Laudet, P.; Lautier, N.; Lecann, H.; Lefevre, L.; Leruyet, B.; Levacher, P.; Magnan, A.; Mazy, E.; Mertens, F.; Mesnager, J.-M.; Meunier, J.-C.; Michel, J.-P.; Monjoin, W.; Naudet, D.; Nguyen-Kim, K.; Orcesi, J.-L.; Ottacher, H.; Perez, R.; Peter, G.; Plasson, P.; Plesseria, J.-Y.; Pontet, B.; Pradines, A.; Quentin, C.; Reynaud, J.-L.; Rolland, G.; Rollenhagen, F.; Romagnan, R.; Russ, N.; Schmidt, R.; Schwartz, N.; Sebbag, I.; Sedes, G.; Smit, H.; Steller, M. B.; Sunter, W.; Surace, C.; Tello, M.; Tiphène, D.; Toulouse, P.; Ulmer, B.; Vandermarcq, O.; Vergnault, E.; Vuillemin, A.; Zanatta, P.

    2009-10-01

    Aims: We report the discovery of very shallow (Δ F/F ≈ 3.4× 10-4), periodic dips in the light curve of an active V = 11.7 G9V star observed by the CoRoT satellite, which we interpret as caused by a transiting companion. We describe the 3-colour CoRoT data and complementary ground-based observations that support the planetary nature of the companion. Methods: We used CoRoT colours information, good angular resolution ground-based photometric observations in- and out- of transit, adaptive optics imaging, near-infrared spectroscopy, and preliminary results from radial velocity measurements, to test the diluted eclipsing binary scenarios. The parameters of the host star were derived from optical spectra, which were then combined with the CoRoT light curve to derive parameters of the companion. Results: We examined all conceivable cases of false positives carefully, and all the tests support the planetary hypothesis. Blends with separation >0.40´´or triple systems are almost excluded with a 8 × 10-4 risk left. We conclude that, inasmuch we have been exhaustive, we have discovered a planetary companion, named CoRoT-7b, for which we derive a period of 0.853 59 ± 3 × 10-5 day and a radius of Rp = 1.68 ± 0.09 R_Earth. Analysis of preliminary radial velocity data yields an upper limit of 21 M_Earth for the companion mass, supporting the finding. Conclusions: CoRoT-7b is very likely the first Super-Earth with a measured radius. This object illustrates what will probably become a common situation with missions such as Kepler, namely the need to establish the planetary origin of transits in the absence of a firm radial velocity detection and mass measurement. The composition of CoRoT-7b remains loosely constrained without a precise mass. A very high surface temperature on its irradiated face, ≈1800-2600 K at the substellar point, and a very low one, ≈50 K, on its dark face assuming no atmosphere, have been derived. The CoRoT space mission, launched on 27

  15. A Potent Derivative of Indolizino[6,7-b]Indole for Treatment of Human Non–Small Cell Lung Cancer Cells12

    PubMed Central

    Chen, Chi-Wei; Wu, Ming-Hsi; Chen, Yi-Fan; Yen, Tsai-Yi; Lin, Yi-Wen; Chao, Shu-Hsin; Tala, Satishkumar; Tsai, Tung-Hu; Su, Tsann-Long; Lee, Te-Chang

    2016-01-01

    The therapeutic effect in non–small cell lung cancer (NSCLC) patients is limited because of intrinsic and acquired resistance. Thus, an unmet need exists for the development of new drugs to improve the therapeutic efficacy in NSCLC patients. In this study, the novel small molecule indolizino[6,7-b]indole derivative BO-1978 was selected to evaluate its therapeutic effects on NSCLC and its preclinical toxicity in animal models. An in vitro cytotoxicity assay revealed that BO-1978 significantly suppressed the growth of various NSCLC cell lines with or without mutations in epidermal growth factor receptor (EGFR). Mechanistically, we demonstrated that BO-1978 exhibited multiple modes of action, including inhibition of topoisomerase I/II and induction of DNA cross-linking. Treatment of NSCLC cells with BO-1978 caused DNA damage, disturbed cell cycle progression, and triggered apoptotic cell death. Furthermore, BO-1978 significantly suppressed the growth of EGFR wild-type and mutant NSCLC tumors in xenograft tumor and orthotopic lung tumor models with negligible body weight loss. The combination of BO-1978 with gefitinib further suppressed EGFR mutant NSCLC cell growth in xenograft tumor and orthotopic lung tumor models. Preclinical toxicity studies showed that BO-1978 administration did not cause apparent toxicity in mice. Based on its significant therapeutic efficacy and low drug toxicity, BO-1978 is a potential therapeutic agent for treatment of NSCLC. PMID:27108383

  16. N,N'-Bis(2-cyclohexylethyl)naphtho[2,3-b:6,7-b']dithiophene Diimides: Effects of Substituents.

    PubMed

    Nakano, Masahiro; Hashizume, Daisuke; Takimiya, Kazuo

    2016-01-01

    Naphtho[2,3-b:6,7-b']dithiophene-4,5,9,10-tetracarboxylic diimide (NDTI) is a promising electron-deficient building block for n-type organic conductors, and the performance of NDTI-based field-effect transistors (FETs) is largely dependent on the substituents that alter the supramolecular organization in the solid state and, in turn, the intermolecular orbital overlap. For this reason, the rational selection of substituent on imide nitrogen atoms and/or thiophene α-positions is the key to developing superior n-type organic semiconductors. We here report new NDTI derivatives having N-(2-cyclohexylethyl) groups. Despite their one-dimensional packing structures in the solid state regardless of the presence or absence of chlorine groups at the thiophene α-positions, their FETs show promising performance with electron mobilities higher than 0.1 cm²·V(-1)·s(-1) under ambient conditions. We also discuss how the cyclohexylethyl groups affect the packing structure in comparison with analogous n-octyl derivatives having the same number of carbon atoms. PMID:27483217

  17. A Potent Derivative of Indolizino[6,7-b]Indole for Treatment of Human Non-Small Cell Lung Cancer Cells.

    PubMed

    Chen, Chi-Wei; Wu, Ming-Hsi; Chen, Yi-Fan; Yen, Tsai-Yi; Lin, Yi-Wen; Chao, Shu-Hsin; Tala, Satishkumar; Tsai, Tung-Hu; Su, Tsann-Long; Lee, Te-Chang

    2016-04-01

    The therapeutic effect in non-small cell lung cancer (NSCLC) patients is limited because of intrinsic and acquired resistance. Thus, an unmet need exists for the development of new drugs to improve the therapeutic efficacy in NSCLC patients. In this study, the novel small molecule indolizino[6,7-b]indole derivative BO-1978 was selected to evaluate its therapeutic effects on NSCLC and its preclinical toxicity in animal models. An in vitro cytotoxicity assay revealed that BO-1978 significantly suppressed the growth of various NSCLC cell lines with or without mutations in epidermal growth factor receptor (EGFR). Mechanistically, we demonstrated that BO-1978 exhibited multiple modes of action, including inhibition of topoisomerase I/II and induction of DNA cross-linking. Treatment of NSCLC cells with BO-1978 caused DNA damage, disturbed cell cycle progression, and triggered apoptotic cell death. Furthermore, BO-1978 significantly suppressed the growth of EGFR wild-type and mutant NSCLC tumors in xenograft tumor and orthotopic lung tumor models with negligible body weight loss. The combination of BO-1978 with gefitinib further suppressed EGFR mutant NSCLC cell growth in xenograft tumor and orthotopic lung tumor models. Preclinical toxicity studies showed that BO-1978 administration did not cause apparent toxicity in mice. Based on its significant therapeutic efficacy and low drug toxicity, BO-1978 is a potential therapeutic agent for treatment of NSCLC.

  18. LIN28B suppresses microRNA let-7b expression to promote CD44+/LIN28B+ human pancreatic cancer stem cell proliferation and invasion

    PubMed Central

    Shao, Yebo; Zhang, Lei; Cui, Lei; Lou, Wenhui; Wang, Dansong; Lu, Weiqi; Jin, Dayong; Liu, Te

    2015-01-01

    Although the highly proliferative, migratory, and multi-drug resistant phenotype of human pancreatic cancer stem cells (PCSCs) is well characterized, knowledge of their biological mechanisms is limited. We used CD44 and LIN28B as markers to screen, isolate, and enrich CSCs from human primary pancreatic cancer. Using flow cytometry, we identified a human primary pancreatic cancer cell (PCC) subpopulation expressing high levels of both CD44 and LIN28B. CD44+/LIN28B+ PCSCs expressed high levels of stemness marker genes and possessed higher migratory and invasive ability than CD44-/LIN28B- PCCs. CD44+/LIN28B+ PCSCs were more resistant to growth inhibition induced by the chemotherapeutic drugs cisplatin and gemcitabine hydrochloride, and readily established tumors in vivo in a relatively short time. Moreover, microarray analysis revealed significant differences between the cDNA expression patterns of CD44+/LIN28B+ PCSCs and CD44-/LIN28B- PCCs. Following siRNA interference of endogenous LIN28B gene expression in CD44+/LIN28B+ PCSCs, not only was their proliferation decreased, there was also cell cycle arrest due to suppression of cyclin D1 expression following the stimulation of miRNA let-7b expression. In conclusion, CD44+/LIN28B+ cells, which possess CSC characteristics, can be reliably sorted from human primary PCCs and represent a valuable model for studying cancer cell physiology and multi-drug resistance. PMID:26609473

  19. Electronic structure, magnetic properties, and stability of the binary and ternary carbides (Fe,Cr)3C and (Fe,Cr)7C3

    NASA Astrophysics Data System (ADS)

    Konyaeva, M. A.; Medvedeva, N. I.

    2009-10-01

    The structural, electronic, and magnetic properties of the binary and ternary carbides (Fe,Cr)3C and (Fe,Cr)7C3 have been investigated within the ab initio density functional theory. The crystal structure of the binary carbides has been optimized and the preferred positions for replacement of chromium or iron impurities in the corresponding carbides have been determined. The changes in the electronic structure and magnetic properties have been investigated, the formation energies of the ternary carbides as functions of the impurity concentrations have been calculated, and conclusions have been drawn regarding the influence of the impurity on the stability of the carbides under investigation.

  20. Autosomal dominant familial spastic paraplegia: reduction of the FSP1 candidate region on chromosome 14q to 7 cM and locus heterogeneity.

    PubMed Central

    Gispert, S; Santos, N; Damen, R; Voit, T; Schulz, J; Klockgether, T; Orozco, G; Kreuz, F; Weissenbach, J; Auburger, G

    1995-01-01

    Three large pedigrees of German descent with autosomal dominant "pure" familial spastic paraplegia (FSP) were characterized clinically and genetically. Haplotype and linkage analyses, with microsatellites covering the FSP region on chromosome 14q (locus FSP1), were performed. In pedigree W, we found a haplotype that cosegregates with the disease and observed three crossing-over events, reducing the FSP1 candidate region to 7 cM; in addition, the observation of apparent anticipation in this family suggests a trinucleotide repeat expansion as the mutation. In pedigrees D and S, the gene locus could be excluded from the whole FSP1 region, confirming the locus heterogeneity of autosomal dominant FSP. PMID:7825576

  1. Formation of linearly linked Fe clusters on Si(111)-7 × 7-C2H5OH surface

    PubMed Central

    2014-01-01

    The Fe atoms were deposited on the Si(111)-7 × 7 surface, which has been saturated with the C2H5OH molecules. Then, the Fe clusters were formed on Si(111)-7 × 7-C2H5OH surface and in situ observed by the scanning tunneling microscopy (STM). The STM images showed that with the increase of Fe clusters, the size of clusters was about 5 nm and they self-assembled in straightly linked chain crossing the step to lower or upper terrace. X-ray photoelectron spectroscopy (XPS) was in situ carried out on the surface of Fe/Si(111)-7 × 7-C2H5OH samples before and after the introduction of thin air (4.5 × 10-2 Langmuir) into the STM chamber. The XPS results showed that the Fe clusters are stable in the abovementioned thin air condition at room temperature. Based on the STM and XPS results, the driving force making one-dimensional straightly linked chain structure might be the magnetic force of the Fe clusters. The formation of straightly linked Fe clusters chains suggests the formation of single magnetic domain Fe clusters. PACS 07.79.Cz, 81.15.-z, 75.75.Fk PMID:25170327

  2. Real-time observation of the swelling and hydrolysis of a single crystalline cellulose fiber catalyzed by cellulase 7B from Trichoderma reesei.

    PubMed

    Wang, Jingpeng; Quirk, Amanda; Lipkowski, Jacek; Dutcher, John R; Hill, Christopher; Mark, Adam; Clarke, Anthony J

    2012-06-26

    The biodegradation of cellulose involves the enzymatic action of cellulases (endoglucanases), cellobiohydrolases (exoglucanases), and β-glucosidases that act synergistically. The rate and efficiency of enzymatic hydrolysis of crystalline cellulose in vitro decline markedly with time, limiting the large-scale, cost-effective production of cellulosic biofuels. Several factors have been suggested to contribute to this phenomenon, but there is considerable disagreement regarding the relative importance of each. These earlier investigations were hampered by the inability to observe the disruption of crystalline cellulose and its subsequent hydrolysis directly. Here, we show the application of high-resolution atomic force microscopy to observe the swelling of a single crystalline cellulose fiber and its-hydrolysis in real time directly as catalyzed by a single cellulase, the industrially important cellulase 7B from Trichoderma reesei. Volume changes, the root-mean-square roughness, and rates of hydrolysis of the surfaces of single fibers were determined directly from the images acquired over time. Hydrolysis dominated the early stage of the experiment, and swelling dominated the later stage. The high-resolution images revealed that the combined action of initial hydrolysis followed by swelling exposed individual microfibrils and bundles of microfibrils, resulting in the loosening of the fiber structure and the exposure of microfibrils at the fiber surface. Both the hydrolysis and swelling were catalyzed by the native cellulase; under the same conditions, its isolated carbohydrate-binding module did not cause changes to crystalline cellulose. We anticipate that the application of our AFM-based analysis on other cellulolytic enzymes, alone and in combination, will provide significant insight into the process of cellulose biodegradation and greatly facilitate its application for the efficient and economical production of cellulosic ethanol.

  3. Substrate-Induced Transcriptional Activation of the MoCel7C Cellulase Gene Is Associated with Methylation of Histone H3 at Lysine 4 in the Rice Blast Fungus Magnaporthe oryzae

    PubMed Central

    Vu, Ba Van; Pham, Kieu Thi Minh

    2013-01-01

    The mechanisms involved in substrate-dependent regulation of a Magnaporthe oryzae gene encoding a cellulase which we designate MoCel7C (MGG_14954) were investigated. The levels of MoCel7C transcript were dramatically increased more than 1,000-fold, 16 to 24 h after transfer to a medium containing 2% carboxymethylcellulose (CMC), while levels were very low or undetectable in conventional rich medium. Green fluorescent protein reporter assays showed that the MoCel7C promoter was activated by cello-oligosaccharides larger than a pentamer. CMC-induced activation of the MoCel7C promoter was suppressed by glucose and cellobiose. Chromatin immunoprecipitation assays revealed that histone H3 methylation on lysine 4 (H3K4) at the MoCel7C locus was associated with activation of the gene by CMC. Consistently, CMC-induced MoCel7C gene activation was drastically diminished in a knockout (KO) mutant of the MoSET1 gene, which encodes a histone lysine methyltransferase that catalyzes H3K4 methylation in M. oryzae. Interestingly, however, MoCel7C transcript levels under noninducing conditions were significantly increased in the MoSET1 KO mutant, suggesting that MoSET1 directly or indirectly plays a role in both activation and suppression of the MoCel7C gene in response to environmental signals. In addition, gene expression and silencing vectors using the MoCel7C promoter were constructed. PMID:23995923

  4. Disruption of copper homeostasis due to a mutation of Atp7a delays the onset of prion disease

    PubMed Central

    Siggs, Owen M.; Cruite, Justin T.; Du, Xin; Rutschmann, Sophie; Masliah, Eliezer; Beutler, Bruce; Oldstone, Michael B. A.

    2012-01-01

    Copper influences the pathogenesis of prion disease, but whether it is beneficial or detrimental remains controversial. Copper homeostasis is also essential for normal physiology, as highlighted by the spectrum of diseases caused by disruption of the copper transporting enzymes ATP7A and ATP7B. Here, by using a forward genetics approach in mice, we describe the isolation of three alleles of Atp7a, each with different phenotypic consequences. The mildest of the three, Atp7abrown, was insufficient to cause lethality in hemizygotes or mottling of the coat in heterozygotes, but did lead to coat hypopigmentation and reduced copper content in the brains of hemizygous males. When challenged with Rocky Mountain Laboratory scrapie, the onset of prion disease was delayed in Atp7abrown mice, and significantly less proteinase-resistant prion protein was found in the brains of moribund Atp7abrown mice compared with WT littermates. Our results establish that ATP7A-mediated copper homeostasis is important for the formation of pathogenic proteinase-resistant prion protein. PMID:22869751

  5. BnaC9.SMG7b Functions as a Positive Regulator of the Number of Seeds per Silique in Brassica napus by Regulating the Formation of Functional Female Gametophytes.

    PubMed

    Li, Shipeng; Chen, Lei; Zhang, Liwu; Li, Xi; Liu, Ying; Wu, Zhikun; Dong, Faming; Wan, Lili; Liu, Kede; Hong, Dengfeng; Yang, Guangsheng

    2015-12-01

    Number of seeds per silique (NSS) is an important determinant of seed yield potential in Brassicaceae crops, and it is controlled by naturally occurring quantitative trait loci. We previously mapped a major quantitative trait locus, qSS.C9, on the C9 chromosome that controls NSS in Brassica napus. To gain a better understanding of how qSS.C9 controls NSS in B. napus, we isolated this locus through a map-based cloning strategy. qSS.C9 encodes a predicted small protein with 119 amino acids, designated as BnaC9.SMG7b, that shows homology with the Ever ShorterTelomere1 tertratricopeptide repeats and Ever Shorter Telomere central domains of Arabidopsis (Arabidopsis thaliana) SUPPRESSOR WITH MORPHOGENETIC EFFECTS ON GENITALIA7 (SMG7). BnaC9.SMG7b plays a role in regulating the formation of functional female gametophyte, thus determining the formation of functional megaspores and then mature ovules. Natural loss or artificial knockdown of BnaC9.SMG7b significantly reduces the number of functional ovules per silique and thus, results in decreased seed number, indicating that qSS.C9 is a positive regulator of NSS in B. napus. Sequence and function analyses show that BnaC9.SMG7b experiences a subfunctionalization process that causes loss of function in nonsense-mediated mRNA decay, such as in Arabidopsis SMG7. Haplotype analysis in 84 accessions showed that the favorable BnaC9.SMG7b alleles are prevalent in modern B. napus germplasms, suggesting that this locus has been a major selection target of B. napus improvement. Our results represent the first step toward unraveling the molecular mechanism that controls the natural variation of NSS in B. napus.

  6. Effective treatment of a murine model of adult T-cell leukemia using 211At-7G7/B6 and its combination with unmodified anti-Tac (daclizumab) directed toward CD25.

    PubMed

    Zhang, Zhuo; Zhang, Meili; Garmestani, Kayhan; Talanov, Vladimir S; Plascjak, Paul S; Beck, Barbara; Goldman, Carolyn; Brechbiel, Martin W; Waldmann, Thomas A

    2006-08-01

    Adult T-cell leukemia (ATL) consists of an overabundance of T cells, which express CD25. Therapeutic efficacy of astatine-211 ((211)At)-labeled murine monoclonal antibody 7G7/B6 alone and in combination with daclizumab was evaluated in nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice given injections of MET-1 human T-cell leukemia cells. Daclizumab and 7G7/B6 are directed toward different epitopes of CD25. Either a single dose of 12 microCi (0.444 MBq) (211)At-7G7/B6 per mouse given intravenously or receptor-saturating doses of daclizumab given at 100 microg weekly for 4 weeks intravenously inhibited tumor growth as monitored by serum levels of human beta-2 microglobulin (beta(2)mu) and by prolonged survival of leukemia-bearing mice compared with the control groups (P < .001). The combination of 2 agents enhanced the antitumor effect when compared with groups treated with 12 microCi (0.444 MBq) of (211)At-7G7/B6 (P < .05) or daclizumab alone (P < .05). The median survival duration of the PBS group was 62.6 days and 61.5 days in the radiolabeled nonspecific antibody (211)At-11F11-treated group. In contrast, 91% of mice in the combination group survived through day 94. These results that demonstrate a significantly improved therapeutic efficacy by combining (211)At-7G7/B6 with daclizumab support a clinical trial of this regimen in patients with ATL. PMID:16569769

  7. Novel BCOR-MAML3 and ZC3H7B-BCOR Gene Fusions in Undifferentiated Small Blue Round Cell Sarcomas.

    PubMed

    Specht, Katja; Zhang, Lei; Sung, Yun-Shao; Nucci, Marisa; Dry, Sarah; Vaiyapuri, Sumathi; Richter, Gunther H S; Fletcher, Christopher D M; Antonescu, Cristina R

    2016-04-01

    Small blue round cell tumors (SBRCTs) are a heterogenous group of tumors that are difficult to diagnose because of overlapping morphologic, immunohistochemical, and clinical features. About two-thirds of EWSR1-negative SBRCTs are associated with CIC-DUX4-related fusions, whereas another small subset shows BCOR-CCNB3 X-chromosomal paracentric inversion. Applying paired-end RNA sequencing to an SBRCT index case of a 44-year-old man, we identified a novel BCOR-MAML3 chimeric fusion, which was validated by reverse transcription polymerase chain reaction and fluorescence in situ hybridization techniques. We then screened a total of 75 SBRCTs lacking EWSR1, FUS, SYT, CIC, and BCOR-CCNB3 abnormalities for BCOR break-apart probes by fluorescence in situ hybridization to detect potential recurrent BCOR gene rearrangements outside the typical X-chromosomal inversion. Indeed, 8/75 (11%) SBRCTs showed distinct BCOR gene rearrangements, with 2 cases each showing either a BCOR-MAML3 or the alternative ZC3H7B-BCOR fusion, whereas no fusion partner was detected in the remaining 4 cases. Gene expression of the BCOR-MAML3-positive index case showed a distinct transcriptional profile with upregulation of HOX-gene signature, compared with classic Ewing's sarcoma or CIC-DUX4-positive SBRCTs. The clinicopathologic features of the SBRCTs with alternative BCOR rearrangements were also compared with a group of BCOR-CCNB3 inversion-positive cases, combining 11 from our files with a meta-analysis of 42 published cases. The BCOR-CCNB3-positive tumors occurred preferentially in children and in bone, in contrast to alternative BCOR-rearranged SBRCTs, which presented in young adults, with a variable anatomic distribution. Furthermore, BCOR-rearranged tumors often displayed spindle cell areas, either well defined in intersecting fascicles or blending with the round cell component, which appears distinct from most other fusion-positive SBRCTs and shares histologic overlap with poorly

  8. Immunogenicity, reactogenicity, and safety of a P1.7b,4 strain-specific serogroup B meningococcal vaccine given to preteens.

    PubMed

    Hosking, Jamie; Rasanathan, Kumanan; Mow, Florina Chan; Jackson, Catherine; Martin, Diana; O'Hallahan, Jane; Oster, Philipp; Ypma, Ellen; Reid, Stewart; Aaberge, Ingeborg; Crengle, Sue; Stewart, Joanna; Lennon, Diana

    2007-11-01

    New Zealand (NZ) has experienced a Neisseria meningitidis serogroup B epidemic since 1991. MeNZB, a strain-specific outer membrane vesicle vaccine made using an NZ epidemic strain isolate, NZ98/254 (B:4:P1.7b,4), from two manufacturing sites, the Norwegian Institute of Public Health (NIPH) and Chiron Vaccines (CV; now Novartis), was evaluated for safety, immunogenicity, and reactogenicity in this observer-blind trial with 8- to 12-year-old children. In year 1, cohort A (n = 302) was randomized 4:1 for receipt of NIPH-MeNZB or MenBvac (Norwegian parent vaccine strain 44/76; B:15:P1.7,16). In year 2, cohort B (n = 313) was randomized 4:1 for receipt of CV-MeNZB or NIPH-MeNZB. Participants all received three vaccinations 6 weeks apart. Local and systemic reactions were monitored for 7 days. Seroresponse was defined as a fourfold or greater rise in the serum bactericidal antibody titer from the baseline titer as measured by a serum bactericidal assay. Those with baseline titers of <1:4 required titers of >/=1:8 to serorespond. Intention-to-treat (ITT) and per protocol (PP) analyses are presented. In cohort A, 74% (ITT) and 73% (PP) of NIPH-MeNZB recipients demonstrated seroresponses against NZ98/254 after three doses, versus 32% (ITT and PP) of MenBvac recipients. In cohort B, seroresponses against NZ98/254 after three doses occurred in 79% (ITT and PP) of CV-MeNZB versus 75% (ITT) and 76% (PP) of NIPH-MeNZB recipients. Vaccines were tolerable, with no vaccine-related serious adverse events. In conclusion, the NZ strain meningococcal B vaccine (MeNZB) from either manufacturing site was immunogenic against New Zealand epidemic vaccine strain meningococci with no safety concerns when given in three doses to these 8- to 12-year-old children.

  9. Immunogenicity, reactogenicity, and safety of a P1.7b,4 strain-specific serogroup B meningococcal vaccine given to preteens.

    PubMed

    Hosking, Jamie; Rasanathan, Kumanan; Mow, Florina Chan; Jackson, Catherine; Martin, Diana; O'Hallahan, Jane; Oster, Philipp; Ypma, Ellen; Reid, Stewart; Aaberge, Ingeborg; Crengle, Sue; Stewart, Joanna; Lennon, Diana

    2007-11-01

    New Zealand (NZ) has experienced a Neisseria meningitidis serogroup B epidemic since 1991. MeNZB, a strain-specific outer membrane vesicle vaccine made using an NZ epidemic strain isolate, NZ98/254 (B:4:P1.7b,4), from two manufacturing sites, the Norwegian Institute of Public Health (NIPH) and Chiron Vaccines (CV; now Novartis), was evaluated for safety, immunogenicity, and reactogenicity in this observer-blind trial with 8- to 12-year-old children. In year 1, cohort A (n = 302) was randomized 4:1 for receipt of NIPH-MeNZB or MenBvac (Norwegian parent vaccine strain 44/76; B:15:P1.7,16). In year 2, cohort B (n = 313) was randomized 4:1 for receipt of CV-MeNZB or NIPH-MeNZB. Participants all received three vaccinations 6 weeks apart. Local and systemic reactions were monitored for 7 days. Seroresponse was defined as a fourfold or greater rise in the serum bactericidal antibody titer from the baseline titer as measured by a serum bactericidal assay. Those with baseline titers of <1:4 required titers of >/=1:8 to serorespond. Intention-to-treat (ITT) and per protocol (PP) analyses are presented. In cohort A, 74% (ITT) and 73% (PP) of NIPH-MeNZB recipients demonstrated seroresponses against NZ98/254 after three doses, versus 32% (ITT and PP) of MenBvac recipients. In cohort B, seroresponses against NZ98/254 after three doses occurred in 79% (ITT and PP) of CV-MeNZB versus 75% (ITT) and 76% (PP) of NIPH-MeNZB recipients. Vaccines were tolerable, with no vaccine-related serious adverse events. In conclusion, the NZ strain meningococcal B vaccine (MeNZB) from either manufacturing site was immunogenic against New Zealand epidemic vaccine strain meningococci with no safety concerns when given in three doses to these 8- to 12-year-old children. PMID:17898183

  10. Identification of a 7-cM region of frequent allelic loss on chromosome band 16p13.3 that is specifically associated with anaplastic thyroid carcinoma.

    PubMed

    Kadota, M; Tamaki, Y; Sakita, I; Komoike, Y; Miyazaki, M; Ooka, M; Masuda, N; Fujiwara, Y; Ohnishi, T; Tomita, N; Sekimoto, M; Ohue, M; Ikeda, T; Kobayashi, T; Horii, A; Monden, M

    2000-01-01

    A total of 17 primary thyroid cancer specimens including seven anaplastic cancers, two papillary cancers adjacent to the anaplastic cancers, and eight papillary cancers were analyzed for loss of heterozygosity (LOH) on chromosome arm 16p. All tumors of anaplastic cancer showed LOHs at one or more loci, and a 7-cM region of the smallest deleted region was found on 16p13.3 between D16S423 and D16S406. This LOH was specifically found in the anaplastic cancer and not in the papillary thyroid cancer. Our present results suggest localization of the putative tumor suppressor gene on 16p13.3, which is likely to play an important role in the anaplastic transformation of thyroid cancer.

  11. LBT/LUCIFER view of star-forming galaxies in the cluster 7C 1756+6520 at z ˜ 1.4

    NASA Astrophysics Data System (ADS)

    Magrini, Laura; Sommariva, Veronica; Cresci, Giovanni; Sani, Eleonora; Galametz, Audrey; Mannucci, Filippo; Petropoulou, Vasiliki; Fumana, Marco

    2012-10-01

    Galaxy clusters are key places to study the contribution of nature (i.e. mass and morphology) and nurture (i.e. environment) in the formation and evolution of galaxies. Recently, a number of clusters at z > 1, i.e. corresponding to the first epochs of the cluster formation, have been discovered and confirmed spectroscopically. We present new observations obtained with the LBT Near Infrared Spectroscopic Utility with Camera and Integral Field Unit for Extragalactic Research (LUCIFER) spectrograph at Large Binocular Telescope (LBT) of a sample of star-forming galaxies associated with a large-scale structure around the radio galaxy 7C 1756+6520 at z = 1.42. Combining our spectroscopic data and the literature photometric data, we derived some of the properties of these galaxies: star formation rate, metallicity and stellar mass. With the aim of analysing the effect of the cluster environment on galaxy evolution, we have located the galaxies in the plane of the so-called fundamental metallicity relation (FMR), which is known not to evolve with redshift up to z = 2.5 for field galaxies, but it is still unexplored in rich environments at low and high redshifts. We found that the properties of the galaxies in the cluster 7C 1756+6520 are compatible with the FMR which suggests that the effect of the environment on galaxy metallicity at this early epoch of cluster formation is marginal. As a side study, we also report the spectroscopic analysis of a bright active galactic nucleus, belonging to the cluster, which shows a significant outflow of gas.

  12. 7 CFR 201.7a - Treated seed.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 3 2011-01-01 2011-01-01 false Treated seed. 201.7a Section 201.7a Agriculture..., Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE (CONTINUED) FEDERAL SEED ACT FEDERAL SEED ACT REGULATIONS Records for Agricultural and Vegetable Seeds § 201.7a Treated seed. The complete record for...

  13. 7 CFR 201.7a - Treated seed.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 3 2013-01-01 2013-01-01 false Treated seed. 201.7a Section 201.7a Agriculture..., Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE (CONTINUED) FEDERAL SEED ACT FEDERAL SEED ACT REGULATIONS Records for Agricultural and Vegetable Seeds § 201.7a Treated seed. The complete record for...

  14. 7 CFR 201.7a - Treated seed.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Treated seed. 201.7a Section 201.7a Agriculture..., Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE (CONTINUED) FEDERAL SEED ACT FEDERAL SEED ACT REGULATIONS Records for Agricultural and Vegetable Seeds § 201.7a Treated seed. The complete record for...

  15. 7 CFR 201.7a - Treated seed.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 3 2012-01-01 2012-01-01 false Treated seed. 201.7a Section 201.7a Agriculture..., Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE (CONTINUED) FEDERAL SEED ACT FEDERAL SEED ACT REGULATIONS Records for Agricultural and Vegetable Seeds § 201.7a Treated seed. The complete record for...

  16. 7 CFR 201.7a - Treated seed.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 3 2014-01-01 2014-01-01 false Treated seed. 201.7a Section 201.7a Agriculture..., Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE (CONTINUED) FEDERAL SEED ACT FEDERAL SEED ACT REGULATIONS Records for Agricultural and Vegetable Seeds § 201.7a Treated seed. The complete record for...

  17. Function of Partially Duplicated Human α7 Nicotinic Receptor Subunit CHRFAM7A Gene

    PubMed Central

    de Lucas-Cerrillo, Ana M.; Maldifassi, M. Constanza; Arnalich, Francisco; Renart, Jaime; Atienza, Gema; Serantes, Rocío; Cruces, Jesús; Sánchez-Pacheco, Aurora; Andrés-Mateos, Eva; Montiel, Carmen

    2011-01-01

    The neuronal α7 nicotinic receptor subunit gene (CHRNA7) is partially duplicated in the human genome forming a hybrid gene (CHRFAM7A) with the novel FAM7A gene. The hybrid gene transcript, dupα7, has been identified in brain, immune cells, and the HL-60 cell line, although its translation and function are still unknown. In this study, dupα7 cDNA has been cloned and expressed in GH4C1 cells and Xenopus oocytes to study the pattern and functional role of the expressed protein. Our results reveal that dupα7 transcript was natively translated in HL-60 cells and heterologously expressed in GH4C1 cells and oocytes. Injection of dupα7 mRNA into oocytes failed to generate functional receptors, but when co-injected with α7 mRNA at α7/dupα7 ratios of 5:1, 2:1, 1:1, 1:5, and 1:10, it reduced the nicotine-elicited α7 current generated in control oocytes (α7 alone) by 26, 53, 75, 93, and 94%, respectively. This effect is mainly due to a reduction in the number of functional α7 receptors reaching the oocyte membrane, as deduced from α-bungarotoxin binding and fluorescent confocal assays. Two additional findings open the possibility that the dominant negative effect of dupα7 on α7 receptor activity observed in vitro could be extrapolated to in vivo situations. (i) Compared with α7 mRNA, basal dupα7 mRNA levels are substantial in human cerebral cortex and higher in macrophages. (ii) dupα7 mRNA levels in macrophages are down-regulated by IL-1β, LPS, and nicotine. Thus, dupα7 could modulate α7 receptor-mediated synaptic transmission and cholinergic anti-inflammatory response. PMID:21047781

  18. The family II carbohydrate-binding module of xylanase CflXyn11A from Cellulomonas flavigena increases the synergy with cellulase TrCel7B from Trichoderma reesei during the hydrolysis of sugar cane bagasse.

    PubMed

    Pavón-Orozco, Patricia; Santiago-Hernández, Alejandro; Rosengren, Anna; Hidalgo-Lara, María Eugenia; Stålbrand, Henrik

    2012-01-01

    Synergy between Cellulomonas flavigena xylanase CflXyn11A and Trichoderma reesei endoglucanase TrCel7B was assessed during hydrolysis of alkaline pretreated sugar cane bagasse (SCB) after 12-48 h, applying the individual enzymes and mixtures of the enzymes. A high degree of synergy (6.3) between CflXyn11A and TrCel7B in hydrolysis of SCB was observed after 12h in the equimolar mixture. A threefold decrease in the degree of synergy was observed with TrCel7B and the catalytic module of CflXyn11A; suggesting an important role played by the carbohydrate-binding module of CflXyn11A (CflXyn11A-CBM) in the observed synergy. Affinity electrophoresis and binding assays showed that CflXyn11A-CBM binds to xylans and to a lesser extent to cellulose. Our results suggest that synergy is more pronounced at early stages of hydrolysis. Furthermore, for the first time it is described that a CBM carried by a xylanase significantly enhances the synergy with a cellulase (threefold increase in synergy).

  19. Acidities of closo-1-COOH-1,7-C2B10H11 and amino acids based on icosahedral carbaboranes.

    PubMed

    Dávalos, Juan Z; González, Javier; Ramos, Rocío; Hnyk, Drahomír; Holub, Josef; Santaballa, J Arturo; Canle-L, Moisés; Oliva, Josep M

    2014-04-17

    Carborane clusters are not found in Nature and are exclusively man-made. In this work we study, both experimentally and computationally, the gas-phase acidity (measured GA = 1325 kJ·mol(-1), computed GA = 1321 kJ·mol(-1)) and liquid-phase acidity (measured pKa = 2.00, computed pKa = 1.88) of the carborane acid closo-1-COOH-1,7-C2B10H11. The experimental gas-phase acidity was determined with electrospray tandem mass spectrometry (ESI/MS), by using the extended Cooks kinetic method (EKM). Given the similar spatial requirements of the title icosahedral cage and benzene and the known importance of aminoacids as a whole, such a study is extended, within an acid-base context, to corresponding ortho, meta, and para amino acids derived from icosahedral carborane cages, 1-COOH-n-NH2-1, n-R with {R = C2B10H10, n = 2, 7, 12}, and from benzene {R = C6H4, n = 2, 3, 4}. A remarkable difference is found between the proportion of neutral versus zwitterion structures in water for glycine and the carborane derived amino acids.

  20. GC×GC measurements of C7-C11 aromatic and n-alkane hydrocarbons on Crete, in air from Eastern Europe during the MINOS campaign

    NASA Astrophysics Data System (ADS)

    Xu, X.; Williams, J.; Plass-Dülmer, C.; Berresheim, H.; Salisbury, G.; Lange, L.; Lelieveld, J.

    2003-03-01

    During the Mediterranean Intensive Oxidant Study (MINOS) campaign in August 2001 gas-phase organic compounds were measured using comprehensive two-dimensional gas chromatography (GC×GC) at the Finokalia ground station, Crete. In this paper, C7-C11 aromatic and n-alkane measurements are presented and interpreted. The mean mixing ratios of the hydrocarbons varied from 1+/-1 pptv (i-propylbenzene) to 43+/-36 pptv (toluene). The observed mixing ratios showed strong day-to-day variations and generally higher levels during the first half of the campaign. Mean diel profiles showed maxima at local midnight and late morning, and minima in the early morning and evening. Results from analysis using a simplified box model suggest that both the chemical sink (i.e. reaction with OH) and the variability of source strengths were the causes of the observed variations in hydrocarbon mixing ratios. The logarithms of hydrocarbon concentrations were negatively correlated with the OH concentrations integral over a day prior to the hydrocarbon measurements. Slopes of the regression lines derived from these correlations for different compounds are compared with literature rate constants for their reactions with OH. The slopes for most compounds agree reasonably well with the literature rate constants. A sequential reaction model has been applied to the interpretation of the relationship between ethylbenzene and two of its potential products, i.e. acetophenone and benzeneacetaldehyde. The model can explain the good correlation observed between [acetophenone]/[ethylbenzene] and [benzeneacetaldehyde]/[ethylbenzene]. The model results and field measurements suggest that the reactivity of benzeneacetaldehyde may lie between those of acetophenone and ethylbenzene and that the ratio between yields of acetophenone and benzeneacetaldehyde may be up to 28:1. Photochemical ages of trace gases sampled at Finokalia during the campaign are estimated using the sequential reaction model and related data

  1. GC×GC measurements of C7-C11 aromatic and n-alkane hydrocarbons on Crete, in air from Eastern Europe during the MINOS campaign

    NASA Astrophysics Data System (ADS)

    Xu, X.; Williams, C.; Plass-Dülmer, H.; Berresheim, H.; Salisbury, G.; Lange, L.; Lelieveld, J.

    2003-09-01

    During the Mediterranean Intensive Oxidant Study (MINOS) campaign in August 2001 gas-phase organic compounds were measured using comprehensive two-dimensional gas chromatography (GCxGC) at the Finokalia ground station, Crete. In this paper, C7-C11 aromatic and n-alkane measurements are presented and interpreted. The mean mixing ratios of the hydrocarbons varied from 1±1 pptv (i-propylbenzene) to 43±36 pptv (toluene). The observed mixing ratios showed strong day-to-day variations and generally higher levels during the first half of the campaign. Mean diel profiles showed maxima at local midnight and late morning, and minima in the early morning and evening. Results from analysis using a simplified box model suggest that both the chemical sink (i.e. reaction with OH) and the variability of source strengths were the causes of the observed variations in hydrocarbon mixing ratios. The logarithms of hydrocarbon concentrations were negatively correlated with the OH concentrations integral over a day prior to the hydrocarbon measurements. Slopes of the regression lines derived from these correlations for different compounds are compared with literature rate constants for their reactions with OH. The slopes for most compounds agree reasonably well with the literature rate constants. A sequential reaction model has been applied to the interpretation of the relationship between ethylbenzene and two of its potential products, i.e. acetophenone and benzeneacetaldehyde. The model can explain the good correlation observed between [acetophenone]/[ethylbenzene] and [benzeneacetaldehyde]/[ethylbenzene]. The model results and field measurements suggest that the reactivity of benzeneacetaldehyde may lie between those of acetophenone and ethylbenzene and that the ratio between yields of acetophenone and benzeneacetaldehyde may be up to 28:1. Photochemical ages of trace gases sampled at Finokalia during the campaign are estimated using the sequential reaction model and related data

  2. Discovery of a Neuroprotective Chemical, (S)-N-(3-(3,6-Dibromo-9H-carbazol-9-yl)-2-fluoropropyl)-6-methoxypyridin-2-amine [(−)-P7C3-S243], with Improved Druglike Properties

    PubMed Central

    2015-01-01

    (−)-P7C3-S243 is a neuroprotective aminopropyl carbazole with improved druglike properties compared with previously reported compounds in the P7C3 class. It protects developing neurons in a mouse model of hippocampal neurogenesis and protects mature neurons within the substantia nigra in a mouse model of Parkinson’s disease. A short, enantioselective synthesis provides the neuroprotective agent in optically pure form. It is nontoxic, orally bioavailable, metabolically stable, and able to cross the blood–brain barrier. As such, it represents a valuable lead compound for the development of drugs to treat neurodegenerative diseases and traumatic brain injury. PMID:24697290

  3. ZBTB7A suppresses melanoma metastasis by transcriptionally repressing MCAM

    PubMed Central

    Liu, Xue-Song; Genet, Matthew D; Haines, Jenna E; Mehanna, Elie K; Wu, Shaowei; Chen, Hung-I Harry; Chen, Yidong; Qureshi, Abrar A; Han, Jiali; Chen, Xiang; Fisher, David E; Pandolfi, Pier Paolo; Yuan, Zhi-Min

    2015-01-01

    The excessive metastatic propensity of melanoma makes it the most deadly form of skin cancer, yet the underlying mechanism of metastasis remains elusive. Here, mining of cancer genome datasets discovered a frequent loss of chromosome 19p13.3 and associated down-regulation of the zinc finger transcription factor ZBTB7A in metastatic melanoma. Functional assessment of ZBTB7A-regulated genes identified MCAM, which encodes an adhesion protein key to melanoma metastasis. Using an integrated approach, it is demonstrated that ZBTB7A directly binds to the promoter and transcriptionally represses the expression of MCAM, establishing ZBTB7A as a bona fide transcriptional repressor of MCAM. Consistently, down-regulation of ZBTB7A results in marked upregulation of MCAM and enhanced melanoma cell invasion and metastasis. An inverse correlation of ZBTB7A and MCAM expression in association with melanoma metastasis is further validated with data from analysis of human melanoma specimens. Implications Together these results uncover a previously unrecognized role of ZBTB7A in negative regulation of melanoma metastasis and have important clinical implications. PMID:25995384

  4. Semaphorin 7A Aggravates Pulmonary Inflammation during Lung Injury

    PubMed Central

    Schneider, Mariella; Granja, Tiago Folgosa; Rosenberger, Peter

    2016-01-01

    The extent of pulmonary inflammation during lung injury ultimately determines patient outcome. Pulmonary inflammation is initiated by the migration of neutrophils into the alveolar space. Recent work has demonstrated that the guidance protein semaphorin 7A (SEMA7A) influences the migration of neutrophils into hypoxic tissue sites, yet, its role during lung injury is not well understood. Here, we report that the expression of SEMA7A is induced in vitro through pro-inflammatory cytokines. SEMA7A itself induces the production of pro-inflammatory cytokines in endothelial and epithelial cells, enhancing pulmonary inflammation. The induction of SEMA7A facilitates the transendothelial migration of neutrophils. In vivo, animals with deletion of SEMA7A expression showed reduced signs of pulmonary inflammatory changes following lipopolysaccharide challenge. We define here the role of SEMA7A in the development of lung injury and identify a potential pathway to interfere with these detrimental changes. Future anti-inflammatory strategies for the treatment of lung injury might be based on this finding. PMID:26752048

  5. Waste tank vapor project: Vapor characterization of Tank 241-C-103: Data report for OVS samples collected from Sample Job 7b, Parts I and II, received 5/18/94 and 5/24/94

    SciTech Connect

    Clauss, T.R.; Edwards, J.A.; Fruchter, J.S.

    1994-09-01

    On 5/18/94, Westinghouse Hanford Company (WHC) delivered samples to Pacific Northwest Laboratory (PNL) that were collected from waste Tank 241-C-103 on 5/16/94. These samples were from Sample Job (SJ) 7b, Part 1. On 5/24/94, WHC delivered samples to PNL that were collected from waste Tank 241-C-103 on 5/18/94. These samples were from SJ7b, Part 2. A summary of data derived from the sampling of waste Tank 241-C-103 for gravimetric (H{sub 2}O) and normal paraffin hydrocarbon (NPH) concentrations are shown for SJ7b. Gravimetric analysis was performed on the samples within 24 hours of receipt by PNL. The NPH concentration of 10 samples collected for Part 1 was slightly higher than the average concentration for 15 samples collected in Part 2, 812 ({+-} 133) mg/m{sup 3} and 659 ({+-} 88) mg/m{sup 3}, respectively. The higher concentrations measured in Part 1 samples may be because the samples in Part 1 were collected at a single level, 0.79 meters above the air-liquid interface. Part 2 samples were collected at three different tank levels, 0.79, 2.92, and 5.05 m above the air-liquid interface. In Part 2, the average NPH concentrations for 5 samples collected at each of three levels was similar: 697 (60) mg/m{sup 3} at the low level, 631 (51) mg/m{sup 3} at the mid level, and 651 (134) mg/m{sup 3} at the high level. It is important to note that the measured tridecane to dodecane concentration remained constant in all samples collected in Parts 1 and 2. That ratio is 1.2 {+-} 0.05. This consistent ratio indicates that there were no random analytical biases towards either compound.

  6. Thrombospondin type I domain containing 7A (THSD7A) mediates endothelial cell migration and tube formation.

    PubMed

    Wang, Chieh-Huei; Su, Pei-Tsu; Du, Xiao-Yan; Kuo, Meng-Wei; Lin, Chia-Yi; Yang, Chung-Chi; Chan, Hau-Shien; Chang, Shing-Jyh; Kuo, Calvin; Seo, Kyunga; Leung, Lawrence L; Chuang, Yung-Jen

    2010-03-01

    Angiogenesis is a highly organized process controlled by a series of molecular events. While much effort has been devoted to identifying angiogenic factors and their reciprocal receptors, far less information is available on the molecular mechanisms underlying directed endothelial cell migration. To search for novel proteins that participate in this process, we used the serial analysis of gene expression (SAGE) transcript profiling approach to identify genes that are selectively expressed in endothelial cells (ECs). Two EC SAGE libraries were constructed from human umbilical vein and artery ECs to enable data-mining against other non-ECs. A novel endothelial protein, Thrombospondin Type I Domain Containing 7A (THSD7A), with preferential expression in placenta vasculature and in human umbilical vein endothelial cells (HUVECs) was identified and targeted for further characterization. Overexpression of a THSD7A carboxyl-terminal fragment in HUVECs inhibited cell migration and disrupted tube formation, while suppression of THSD7A expression enhanced HUVEC migration and tube formation. Immunohistological analysis revealed that THSD7A was expressed at the leading edge of migrating HUVECs, and it co-localized with alpha(V)beta(3) integrin and paxillin. This distribution was dispersed from focal adhesions after disruption of the actin cytoskeleton, suggesting the involvement of THSD7A in cytoskeletal organization. Our results show that THSD7A is a novel placenta endothelial protein that mediates EC migration and tube formation, and they highlight its potential as a new target for anti-angiogenic therapy.

  7. DOE evaluation document for DOT 7A Type A packaging

    SciTech Connect

    Edling, D.A.; Hopkins, D.R.; Williams, R.L.

    1987-03-01

    This document is a support document for the ''DOE Evaluation Document for DOT 7A Type A Packaging,'' MLM-3245, March 1987. Provided herein are details concerning the performance requirements specified in 178.350 Specification 7A, General Packaging, Type A. MLM-3245 references appropriate sections in this document. This document does not by itself meet the documentation requirements specified in 49 CFR 173.415 and has compliance value only when used in conjunction with MLM-3245.

  8. Alleviating product inhibition in cellulase enzyme Cel7A

    PubMed Central

    Atreya, Meera E.; Strobel, Kathryn L.

    2015-01-01

    ABSTRACT Enzymes that degrade cellulose into glucose are one of the most expensive components of processes for converting cellulosic biomass to fuels and chemicals. Cellulase enzyme Cel7A is the most abundant enzyme naturally employed by fungi to depolymerize cellulose, and like other cellulases is inhibited by its product, cellobiose. There is thus great economic incentive for minimizing the detrimental effects of product inhibition on Cel7A. In this work, we experimentally generated 10 previously proposed site‐directed mutant Cel7A enzymes expected to have reduced cellobiose binding energies (the majority of mutations were to alanine). We then tested their resilience to cellobiose as well as their hydrolytic activities on microcrystalline cellulose. Although every mutation tested conferred reduced product inhibition (and abolished it for some), our results confirm a trade‐off between Cel7A tolerance to cellobiose and enzymatic activity: Reduced product inhibition was accompanied by lower overall enzymatic activity on crystalline cellulose for the mutants tested. The tempering effect of mutations on inhibition was nearly constant despite relatively large differences in activities of the mutants. Our work identifies an amino acid in the Cel7A product binding site of interest for further mutational studies, and highlights both the challenge and the opportunity of enzyme engineering toward improving product tolerance in Cel7A. Biotechnol. Bioeng. 2016;113: 330–338. © 2015 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc. PMID:26302366

  9. Alleviating product inhibition in cellulase enzyme Cel7A.

    PubMed

    Atreya, Meera E; Strobel, Kathryn L; Clark, Douglas S

    2016-02-01

    Enzymes that degrade cellulose into glucose are one of the most expensive components of processes for converting cellulosic biomass to fuels and chemicals. Cellulase enzyme Cel7A is the most abundant enzyme naturally employed by fungi to depolymerize cellulose, and like other cellulases is inhibited by its product, cellobiose. There is thus great economic incentive for minimizing the detrimental effects of product inhibition on Cel7A. In this work, we experimentally generated 10 previously proposed site-directed mutant Cel7A enzymes expected to have reduced cellobiose binding energies (the majority of mutations were to alanine). We then tested their resilience to cellobiose as well as their hydrolytic activities on microcrystalline cellulose. Although every mutation tested conferred reduced product inhibition (and abolished it for some), our results confirm a trade-off between Cel7A tolerance to cellobiose and enzymatic activity: Reduced product inhibition was accompanied by lower overall enzymatic activity on crystalline cellulose for the mutants tested. The tempering effect of mutations on inhibition was nearly constant despite relatively large differences in activities of the mutants. Our work identifies an amino acid in the Cel7A product binding site of interest for further mutational studies, and highlights both the challenge and the opportunity of enzyme engineering toward improving product tolerance in Cel7A.

  10. Alleviating product inhibition in cellulase enzyme Cel7A.

    PubMed

    Atreya, Meera E; Strobel, Kathryn L; Clark, Douglas S

    2016-02-01

    Enzymes that degrade cellulose into glucose are one of the most expensive components of processes for converting cellulosic biomass to fuels and chemicals. Cellulase enzyme Cel7A is the most abundant enzyme naturally employed by fungi to depolymerize cellulose, and like other cellulases is inhibited by its product, cellobiose. There is thus great economic incentive for minimizing the detrimental effects of product inhibition on Cel7A. In this work, we experimentally generated 10 previously proposed site-directed mutant Cel7A enzymes expected to have reduced cellobiose binding energies (the majority of mutations were to alanine). We then tested their resilience to cellobiose as well as their hydrolytic activities on microcrystalline cellulose. Although every mutation tested conferred reduced product inhibition (and abolished it for some), our results confirm a trade-off between Cel7A tolerance to cellobiose and enzymatic activity: Reduced product inhibition was accompanied by lower overall enzymatic activity on crystalline cellulose for the mutants tested. The tempering effect of mutations on inhibition was nearly constant despite relatively large differences in activities of the mutants. Our work identifies an amino acid in the Cel7A product binding site of interest for further mutational studies, and highlights both the challenge and the opportunity of enzyme engineering toward improving product tolerance in Cel7A. PMID:26302366

  11. Characterization of Vadose Zone Sediment: Borehole C3103 Located in the 216-B-7A Crib Near the B Tank Farm

    SciTech Connect

    Lindenmeier, Clark W.; Serne, R JEFFREY.; Bjornstad, Bruce N.; Last, George V.; Lanigan, David C.; Lindberg, Michael J.; Clayton, Ray E.; Legore, Virginia L.; Kutnyakov, Igor V.; Baum, Steven R.; Geiszler, Keith N.; Valenta, Michelle M.; Vickerman, Tanya S.

    2002-12-01

    This report summarizes data collected from samples in borehole C3103. Borehole C3103 was completed to further characterize the nature and extent of vadose zone contaminants supplied by intentional liquid discharges into the crib 216-B7A/7B between 1954 and 1967. These cribs received dilute waste streams from the bismuth phosphate fuel reprocessing program in the 1950's and decontamination waste in the 1960's. Elevated concentrations of several constituents were primarily measured at different depth intervals. The primary radionuclides present in this borehole are cesium-137 and uranium near the top of the borehole. Chemical characteristics attributed to wastewater-soil interaction at different locations within this zone are elevated pH, sodium, fluoride, carbonate nitrate, and sulphate

  12. G9a/RelB regulates self-renewal and function of colon-cancer-initiating cells by silencing Let-7b and activating the K-RAS/β-catenin pathway.

    PubMed

    Cha, Shih-Ting; Tan, Ching-Ting; Chang, Cheng-Chi; Chu, Chia-Yu; Lee, Wei-Jiunn; Lin, Been-Zen; Lin, Ming-Tsan; Kuo, Min-Liang

    2016-09-01

    Epigenetic reprogramming has been associated with the functional plasticity of cancer-initiating cells (CICs); however, the regulatory pathway has yet to be elucidated. A siRNA screen targeting known epigenetic genes revealed that G9a profoundly impairs the chemo-resistance, self-renewal and metastasis of CICs obtained from patients with colorectal cancer (CRC). Patients with elevated G9a were shown to face a high risk of relapse and poor survival rates. From a mechanistic perspective, G9a binds with and stabilizes RelB, thereby recruiting DNA methyltransferase 3 on the Let-7b promoter and repressing its expression. This leads to the activation of the K-RAS/β-catenin pathway and regulates self-renewal and function of CICs. These findings indicate that the G9a/RelB/Let-7b axis acts as a critical regulator in the maintenance of CIC phenotypes and is strongly associated with negative clinical outcomes. Thus, these findings may have diagnostic as well as therapeutic implications for the treatment of chemotherapy-resistant or metastatic CRC. PMID:27525719

  13. Wilson's disease caused by alternative splicing and Alu exonization due to a homozygous 3039-bp deletion spanning from intron 1 to exon 2 of the ATP7B gene.

    PubMed

    Mameli, Eva; Lepori, Maria Barbara; Chiappe, Francesca; Ranucci, Giusy; Di Dato, Fabiola; Iorio, Raffaele; Loudianos, Georgios

    2015-09-15

    We describe a case of Wilson's disease (WD) diagnosed at 5 years after routine biochemical test showed increased aminotransferases. Mutation analysis of the ATP7B gene revealed a 3039-bp deletion in the homozygous state spanning from the terminal part of intron 1 to nt position 368 of exon 2. This deletion results in the activation of 3 cryptic splice sites: an AG acceptor splice site in nt positions 578-579 producing a different breakpoint and removing the first 577 nts of exon 2, an acceptor and a donor splice site in nt positions 20363-4 and 20456-7, respectively, in intron 1, resulting in the activation of a 94-bp cryptic Alu exon being incorporated into the mature transcript. The resulting alternative transcript contains a TAG stop codon in the first amino acid position of the cryptic exon, likely producing a truncated, non-functional protein. This study shows that intron exonization can also occur in humans through naturally occurring gross deletions. The results suggest that the combination of DNA and RNA analyses can be used for molecular characterization of gross ATP7B deletions, thus improving genetic counseling and diagnosis of WD. Moreover these studies help to better establish new molecular mechanisms producing Wilson's disease.

  14. TTC7A mutations disrupt intestinal epithelial apicobasal polarity

    PubMed Central

    Bigorgne, Amélie E.; Farin, Henner F.; Lemoine, Roxane; Mahlaoui, Nizar; Lambert, Nathalie; Gil, Marine; Schulz, Ansgar; Philippet, Pierre; Schlesser, Patrick; Abrahamsen, Tore G.; Oymar, Knut; Davies, E. Graham; Ellingsen, Christian Lycke; Leteurtre, Emmanuelle; Moreau-Massart, Brigitte; Berrebi, Dominique; Bole-Feysot, Christine; Nischke, Patrick; Brousse, Nicole; Fischer, Alain; Clevers, Hans; de Saint Basile, Geneviève

    2013-01-01

    Multiple intestinal atresia (MIA) is a rare cause of bowel obstruction that is sometimes associated with a combined immunodeficiency (CID), leading to increased susceptibility to infections. The factors underlying this rare disease are poorly understood. We characterized the immunological and intestinal features of 6 unrelated MIA-CID patients. All patients displayed a profound, generalized lymphocytopenia, with few lymphocytes present in the lymph nodes. The thymus was hypoplastic and exhibited an abnormal distribution of epithelial cells. Patients also had profound disruption of the epithelial barrier along the entire gastrointestinal tract. Using linkage analysis and whole-exome sequencing, we identified 10 mutations in tetratricopeptide repeat domain–7A (TTC7A), all of which potentially abrogate TTC7A expression. Intestinal organoid cultures from patient biopsies displayed an inversion of apicobasal polarity of the epithelial cells that was normalized by pharmacological inhibition of Rho kinase. Our data indicate that TTC7A deficiency results in increased Rho kinase activity, which disrupts polarity, growth, and differentiation of intestinal epithelial cells, and which impairs immune cell homeostasis, thereby promoting MIA-CID development. PMID:24292712

  15. 17 CFR 260.7a-4 - Calculation of time.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... eastern daylight-saving time, whichever is in effect at the principal office of the Commission on the date... 17 Commodity and Securities Exchanges 3 2012-04-01 2012-04-01 false Calculation of time. 260.7a-4... time. Saturdays, Sundays and holidays shall be counted in computing the effective date of...

  16. 17 CFR 260.7a-4 - Calculation of time.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... eastern daylight-saving time, whichever is in effect at the principal office of the Commission on the date... 17 Commodity and Securities Exchanges 3 2011-04-01 2011-04-01 false Calculation of time. 260.7a-4... time. Saturdays, Sundays and holidays shall be counted in computing the effective date of...

  17. 17 CFR 260.7a-4 - Calculation of time.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... eastern daylight-saving time, whichever is in effect at the principal office of the Commission on the date... 17 Commodity and Securities Exchanges 3 2010-04-01 2010-04-01 false Calculation of time. 260.7a-4... time. Saturdays, Sundays and holidays shall be counted in computing the effective date of...

  18. 17 CFR 260.7a-4 - Calculation of time.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... eastern daylight-saving time, whichever is in effect at the principal office of the Commission on the date... 17 Commodity and Securities Exchanges 4 2014-04-01 2014-04-01 false Calculation of time. 260.7a-4... time. Saturdays, Sundays and holidays shall be counted in computing the effective date of...

  19. 17 CFR 260.7a-4 - Calculation of time.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... eastern daylight-saving time, whichever is in effect at the principal office of the Commission on the date... 17 Commodity and Securities Exchanges 3 2013-04-01 2013-04-01 false Calculation of time. 260.7a-4... time. Saturdays, Sundays and holidays shall be counted in computing the effective date of...

  20. REORGANIZED SCIENCE CURRICULUM, 7A, GRADE SEVEN SUPPLEMENT.

    ERIC Educational Resources Information Center

    Minneapolis Special School District 1, Minn.

    THE TWELFTH IN A SERIES OF 17 VOLUMES, THIS VOLUME PROVIDES THE SEVENTH GRADE TEACHER WITH A GUIDE TO THE REORGANIZED SCIENCE CURRICULUM OF THE MINNEAPOLIS PUBLIC SCHOOLS. THE MATERIALS ARE INTENDED TO BE AUGMENTED AND REVISED AS THE NEED ARISES. THE SEVENTH GRADE SUPPLEMENT IS IN TWO VOLUMES. VOLUME 7A CONTAINS INTRODUCTORY MATERIAL, A BRIEF…

  1. 77 FR 19531 - 7(a) Loan Program; Eligible Passive Companies

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-02

    ... passive investment or real estate development, or which do not engage in regular and continuous activity... ADMINISTRATION 13 CFR Part 120 RIN 3245-AG48 7(a) Loan Program; Eligible Passive Companies AGENCY: U.S. Small...-guaranteed loan to an Eligible Passive Company. DATES: This rule is effective on May 17, 2012 without...

  2. In situ detection method for obtaining permeability of Fe-based amorphous alloys: ac resistance measurement for Fe{sub 84}Nb{sub 7}B{sub 9}

    SciTech Connect

    Ichitsubo, Tetsu; Matsubara, Eiichiro; Tanaka, Satoshi; Nishiyama, Nobuyuki; Amiya, Kenji

    2005-01-17

    In this letter, we propose an in situ detection method for obtaining permeability of soft magnetic Fe-based amorphous alloys. The temperature dependence of ac resistance was measured at different frequencies during heat treatment of Fe{sub 84}Nb{sub 7}B{sub 9} amorphous alloys. A significant increase in the ac ({approx}1000 kHz) resistance appears at around 920 K during the heating process, which arises from the skin effect caused by a marked increase in sample permeability. This skin effect diminishes when the sample is heated to approximately 1100 K, which corresponds to the decrease in its permeability. Consequently, we note that the ac resistance measurement is useful for quick in situ assessment to achieve the soft magnetic property of an Fe-based amorphous alloy.

  3. Tubby-RFP Balancers for Developmental Analysis: FM7c 2xTb-RFP, CyO 2xTb-RFP and TM3 2xTb-RFP

    PubMed Central

    Pina, Cara; Pignoni, Francesca

    2012-01-01

    We report here the construction of Tubby-RFP balancers for the X, 2nd and 3rd chromosomes of Drosophila melanogaster. The insertion of a 2xTb-RFP transgene on the FM7c, CyO and TM3 balancer chromosomes introduces two easily scorable, dominant, developmental markers. The strong Tb phenotype is visible to the naked eye at the larval L2, L3 and pupal stages. The RFP associated with the cuticle is easily detected at all stages from late embryo to adult with the use of a fluorescence stereomicroscope. The FM7c Bar 2xTb-RFP, CyO Cy 2xTb-RFP and TM3 Sb 2xTb-RFP balancers will greatly facilitate the analysis of lethals and other developmental mutants in L2/L3 larvae and pupae, but also provide coverage of other stages beginning in late embryogenesis through to the adult. PMID:21913310

  4. The Role of Calpain-Myosin 9-Rab7b Pathway in Mediating the Expression of Toll-Like Receptor 4 in Platelets: A Novel Mechanism Involved in α-Granules Trafficking

    PubMed Central

    Tsai, Jui-Chi; Lin, Yi-Wen; Huang, Chun-Yao; Lin, Chih-Yuan; Tsai, Yi-Ting; Shih, Chun-Min; Lee, Chung-Yi; Chen, Yung-Hsiang; Li, Chi-Yuan; Chang, Nen-Chung; Lin, Feng-Yen; Tsai, Chien-Sung

    2014-01-01

    Toll-like receptors (TLRs) plays a critical role in innate immunity. In 2004, Aslam R. and Shiraki R. first determined that murine and human platelets express functional TLRs. Additionally, Andonegui G. demonstrated that platelets express TLR4, which contributes to thrombocytopenia. However, the underlying mechanisms of TLR4 expression by platelets have been rarely explored until now. The aim of this study was to identify the mechanism of TLR4 expression underlying thrombin treatment. The human washed platelets were used in this study. According to flowcytometry and western blot analysis, the surface levels of TLR4 were significantly enhanced in thrombin-activated human platelets and decreased by TMB-8, calpeptin, and U73122, but not Y27632 (a Rho-associated protein kinase ROCK inhibitor) indicating that thrombin-mediated TLR4 expression was modulated by PAR/PLC pathway, calcium and calpain. Co-immunoprecipitation (co-IP) assay demonstrated that the interaction between TLR4 and myosin-9 (a substrate of calpain) was regulated by calpain; cleavage of myosin-9 enhanced TLR4 expression in thrombin treated platelets. Transmission electron microscope data indicated that human platelets used α-granules to control TLR4 expression; the co-IP experiment suggested that myosin-9 did not coordinate with Rab7b to negatively regulate TLR4 trafficking in thrombin treated platelets. In summary, phospholipase Cγ-calpain-myosin 9-Rab7b axis was responsible for the mechanism underlying the regulation of TLR4 containing α-granules trafficking in thrombin-stimulated platelets, which was involved in coagulation. PMID:24489676

  5. SULFATE SOLUBILITY LIMIT VERIFICATION FOR DWPF SLUDGE BATCH 7A

    SciTech Connect

    Billings, A.

    2011-04-19

    During processing at the Defense Waste Processing Facility (DWPF), high sulfate concentrations in the feed are a concern to DWPF as it can lead to the formation of a detrimental, sulfate-rich, molten salt phase on the surface of the glass melt pool. To avoid these issues, a sulfate concentration limit was implemented into the Product Composition Control System (PCCS). Related to SB7a frit development efforts, the Savannah River National Laboratory (SRNL) assessed the viability of using the current 0.6 wt % SO{sub 4}{sup 2-} limit set for SB6 (in glass) and the possibility of increasing the SO{sub 4}{sup 2-} solubility limit in PCCS to account for anticipated sulfur concentrations, targeted waste loadings, and inclusion of secondary streams (e.g., Actinide Removal Process (ARP)) with two recommended frits (Frit 418 and Frit 702) for SB7a processing. For a nominal SB7a blend with a 63 inch SB6 heel remaining in Tank 40 (projection SB7a-63), a 0.60 wt% SO{sub 4}{sup 2-} in glass limit was determined for waste loadings of 34 wt% up to 40 wt% with Frit 418 based on crucible melts with batched chemicals. SRNL also examined the inclusion of ARP for the same blending scenario (SB7a-63-ARP) with Frit 418 and at least a 0.6 wt% SO{sub 4}{sup 2-} level, and waste loadings of 34 wt% to 40 wt% were also acceptable. When a visible yellow and/or white sulfate salt layer was visible on the surface of any cooled glass, it was assumed to have surpassed the solubility limit of SO{sub 4}{sup 2-} for that particular composition. All of the glasses fabricated at these concentrations did not exhibit a sulfate rich salt layer on the surface of the glass melt and retained the majority of the batched SO{sub 4}{sup 2-}. At higher levels of SO{sub 4}{sup 2-} 'spiked' into the projected sludge compositions over the aforementioned interval of waste loadings, with Frit 418, low viscosity sulfur layers were observed on the surface of glass melts which confirm exceeding the solubility limit. The

  6. Crystal structure and electronic properties of the new compounds, U{sub 6}Fe{sub 16}Si{sub 7} and its interstitial carbide U{sub 6}Fe{sub 16}Si{sub 7}C

    SciTech Connect

    Berthebaud, D.; Tougait, O. Potel, M.; Lopes, E.B.; Goncalves, A.P.

    2007-10-15

    The new compounds U6Fe16Si7 and U6Fe16Si7C were prepared by arc-melting and subsequent annealing at 1500 deg. C. Single-crystal X-ray diffraction showed that they crystallize in the cubic space group Fm3-barm (No. 225), with unit-cell parameters at room temperature a=11.7206(5) A for U6Fe16Si7 and a=11.7814(2) A for U6Fe16Si7C. Their crystal structures correspond to ordered variants of the Th6Mn23 type. U6Fe16Si7 adopts the Mg6Cu16Si7 structure type, whereas U6Fe16Si7C crystallizes with a novel 'filled' quaternary variant. The inserted carbon is located in octahedral cages formed by six U atoms, with U-U interatomic distances of 3.509(1) A. Insertion of carbon in the structure of U6Fe16Si7 has a direct influence on the U-Fe and Fe-Fe interatomic distances. The electronic properties of both compounds were investigated by means of DC susceptibility, electrical resistivity and thermopower. U6Fe16Si7 is a Pauli paramagnet. Its electrical resistivity and thermopower point out that it cannot be classified as a simple metal. The magnetic susceptibility of U{sub 6}Fe{sub 16}Si{sub 7}C is best described over the temperature range 100-300 K by using a modified Curie-Weiss law with an effective magnetic moment of 2.3(2) {mu}{sub B}/U, a paramagnetic Weiss temperature, {theta}{sub p}=57(2) K and a temperature-independent term {chi}{sub 0}=0.057(1) emu/mol. Both the electrical resistivity and thermopower reveal metallic behavior. - Graphical abstract: The new compounds U{sub 6}Fe{sub 16}Si{sub 7} and U{sub 6}Fe{sub 16}Si{sub 7}C crystallize with the ternary ordered variant of the Th{sub 6}Mn{sub 23} type, commonly referred as Mg{sub 6}Cu{sub 16}Si7 and with a novel 'filled' variant of this type of structure, respectively.

  7. The Lumenal Loop M672-P707 of the Menkes Protein (ATP7A) Transfers Copper to Peptidylglycine Monooxygenase

    PubMed Central

    Otoikhian, Adenike; Barry, Amanda N.; Mayfield, Mary; Nilges, Mark; Huang, Yiping; Lutsenko, Svetlana; Blackburn, Ninian J.

    2012-01-01

    Copper transfer to cuproproteins located in vesicular compartments of the secretory pathway depends on activity of the copper translocating ATPase (ATP7A or ATP7B) but the mechanism of transfer is largely unexplored. Copper-ATPase ATP7A is unique in having a sequence rich in histidine and methionine residues located on the lumenal side of the membrane. The corresponding fragment binds Cu(I) when expressed as a chimera with a scaffold protein, and mutations or deletions of His and/or Met residues in its sequence inhibit dephosphorylation of the ATPase, a catalytic step associated with copper release. Here we present evidence for a potential role of this lumenal region of ATP7A in copper transfer to cuproenzymes. Both Cu(II) and Cu(I) forms were investigated since the form in which copper is transferred to acceptor proteins is currently unknown. Analysis of Cu(II) using EPR demonstrated that at Cu:P ratios below 1:1, 15N-substituted protein had Cu(II) bound by 4 His residues, but this coordination changed as the Cu(II) to protein ratio increased towards 2:1. XAS confirmed this coordination via analysis of the intensity of outer-shell scattering from imidazole residues. The Cu(II) complexes could be reduced to their Cu(I) counterparts by ascorbate, but here again, as shown by EXAFS and XANES spectroscopy, the coordination was dependent on copper loading. At low copper Cu(I) was bound by a mixed ligand set of His + Met while at higher ratios His coordination predominated. The copper-loaded loop was able to transfer either Cu(II) or Cu(I) to peptidylglycine monooxygenase in the presence of chelating resin, generating catalytically active enzyme in a process that appeared to involve direct interaction between the two partners. The variation of coordination with copper loading suggests copper-dependent conformational change which in turn could act as a signal for regulating copper release by the ATPase pump. PMID:22577880

  8. The lumenal loop M672-P707 of the Menkes protein (ATP7A) transfers copper to peptidylglycine monooxygenase

    SciTech Connect

    Otoikhian, Adenike; Barry, Amanda N.; Mayfield, Mary; Nilges, Mark; Huang, Yiping; Lutsenko, Svetlana; Blackburn, Ninian

    2012-05-14

    Copper transfer to cuproproteins located in vesicular compartments of the secretory pathway depends on activity of the copper translocating ATPase (ATP7A or ATP7B) but the mechanism of transfer is largely unexplored. Copper-ATPase ATP7A is unique in having a sequence rich in histidine and methionine residues located on the lumenal side of the membrane. The corresponding fragment binds Cu(I) when expressed as a chimera with a scaffold protein, and mutations or deletions of His and/or Met residues in its sequence inhibit dephosphorylation of the ATPase, a catalytic step associated with copper release. Here we present evidence for a potential role of this lumenal region of ATP7A in copper transfer to cuproenzymes. Both Cu(II) and Cu(I) forms were investigated since the form in which copper is transferred to acceptor proteins is currently unknown. Analysis of Cu(II) using EPR demonstrated that at Cu:P ratios below 1:1, 15N-substituted protein had Cu(II) bound by 4 His residues, but this coordination changed as the Cu(II) to protein ratio increased towards 2:1. XAS confirmed this coordination via analysis of the intensity of outer-shell scattering from imidazole residues. The Cu(II) complexes could be reduced to their Cu(I) counterparts by ascorbate, but here again, as shown by EXAFS and XANES spectroscopy, the coordination was dependent on copper loading. At low copper Cu(I) was bound by a mixed ligand set of His + Met while at higher ratios His coordination predominated. The copper-loaded loop was able to transfer either Cu(II) or Cu(I) to peptidylglycine monooxygenase in the presence of chelating resin, generating catalytically active enzyme in a process that appeared to involve direct interaction between the two partners. The variation of coordination with copper loading suggests copper-dependent conformational change which in turn could act as a signal for regulating copper release by the ATPase pump.

  9. A novel gene THSD7A is associated with obesity.

    PubMed

    Nizamuddin, S; Govindaraj, P; Saxena, S; Kashyap, M; Mishra, A; Singh, S; Rotti, H; Raval, R; Nayak, J; Bhat, B K; Prasanna, B V; Dhumal, V R; Bhale, S; Joshi, K S; Dedge, A P; Bharadwaj, R; Gangadharan, G G; Nair, S; Gopinath, P M; Patwardhan, B; Kondaiah, P; Satyamoorthy, K; Valiathan, M S; Thangaraj, K

    2015-11-01

    Body mass index (BMI) is a non-invasive measurement of obesity. It is commonly used for assessing adiposity and obesity-related risk prediction. Genetic differences between ethnic groups are important factors, which contribute to the variation in phenotypic effects. India inhabited by the first out-of-Africa human population and the contemporary Indian populations are admixture of two ancestral populations; ancestral north Indians (ANI) and ancestral south Indians (ASI). Although ANI are related to Europeans, ASI are not related to any group outside Indian-subcontinent. Hence, we expect novel genetic loci associated with BMI. In association analysis, we found eight genic SNPs in extreme of distribution (P⩽3.75 × 10(-5)), of which WWOX has already been reported to be associated with obesity-related traits hence excluded from further study. Interestingly, we observed rs1526538, an intronic SNP of THSD7A; a novel gene significantly associated with obesity (P=2.88 × 10(-5), 8.922 × 10(-6) and 2.504 × 10(-9) in discovery, replication and combined stages, respectively). THSD7A is neural N-glycoprotein, which promotes angiogenesis and it is well known that angiogenesis modulates obesity, adipose metabolism and insulin sensitivity, hence our result find a correlation. This information can be used for drug target, early diagnosis of obesity and treatment. PMID:26238973

  10. A novel gene THSD7A is associated with obesity.

    PubMed

    Nizamuddin, S; Govindaraj, P; Saxena, S; Kashyap, M; Mishra, A; Singh, S; Rotti, H; Raval, R; Nayak, J; Bhat, B K; Prasanna, B V; Dhumal, V R; Bhale, S; Joshi, K S; Dedge, A P; Bharadwaj, R; Gangadharan, G G; Nair, S; Gopinath, P M; Patwardhan, B; Kondaiah, P; Satyamoorthy, K; Valiathan, M S; Thangaraj, K

    2015-11-01

    Body mass index (BMI) is a non-invasive measurement of obesity. It is commonly used for assessing adiposity and obesity-related risk prediction. Genetic differences between ethnic groups are important factors, which contribute to the variation in phenotypic effects. India inhabited by the first out-of-Africa human population and the contemporary Indian populations are admixture of two ancestral populations; ancestral north Indians (ANI) and ancestral south Indians (ASI). Although ANI are related to Europeans, ASI are not related to any group outside Indian-subcontinent. Hence, we expect novel genetic loci associated with BMI. In association analysis, we found eight genic SNPs in extreme of distribution (P⩽3.75 × 10(-5)), of which WWOX has already been reported to be associated with obesity-related traits hence excluded from further study. Interestingly, we observed rs1526538, an intronic SNP of THSD7A; a novel gene significantly associated with obesity (P=2.88 × 10(-5), 8.922 × 10(-6) and 2.504 × 10(-9) in discovery, replication and combined stages, respectively). THSD7A is neural N-glycoprotein, which promotes angiogenesis and it is well known that angiogenesis modulates obesity, adipose metabolism and insulin sensitivity, hence our result find a correlation. This information can be used for drug target, early diagnosis of obesity and treatment.

  11. REPORTABLE RADIONUCLIDES IN DWPF SLUDGE BATCH 7A (MACROBATCH 8)

    SciTech Connect

    Reboul, S.; Diprete, D.; Click, D.; Bannochie, C.

    2011-12-20

    The Waste Acceptance Product Specifications (WAPS) 1.2 require that the waste producer 'shall report the curie inventory of radionuclides that have half-lives longer than 10 years and that are, or will be, present in concentrations greater than 0.05 percent of the total inventory for each waste type indexed to the years 2015 and 3115.' As part of the strategy to meet WAPS 1.2, the Defense Waste Processing Facility (DWPF) will report for each waste type all radionuclides that have half-lives longer than 10 years and contribute greater than 0.01 percent of the total curie inventory from the time of production through the 1100 year period from 2015 through 3115. The initial list of radionuclides to be reported is based on the design-basis glass identified in the Waste Form Compliance Plan (WCP) and Waste Form Qualification Report. However, it is required that the list be expanded if other radionuclides with half-lives greater than 10 years are identified that meet the 'greater than 0.01% of the curie inventory' criterion. Specification 1.6 of the WAPS, International Atomic Energy Agency Safeguards Reporting for High Level Waste (HLW), requires that the ratio by weights of the following uranium and plutonium isotopes be reported: U-233, U-234, U-235, U-236, and U-238; and Pu-238, Pu-239, Pu-240, Pu-241, and Pu-242. Therefore, the complete list of reportable radionuclides must also include these sets of U and Pu isotopes - and the U and Pu isotopic mass distributions must be identified. The DWPF receives HLW sludge slurry from Savannah River Site (SRS) Tank 40. For Sludge Batch 7a (SB7a), the waste in Tank 40 contained a blend of the heel from Sludge Batch 6 (SB6) and the Sludge Batch 7 (SB7) material transferred to Tank 40 from Tank 51. This sludge blend is also referred to as Macrobatch 8. Laboratory analyses of a Tank 40 sludge sample were performed to quantify the concentrations of pertinent radionuclides in the SB7a waste. Subsequently, radiological decay and in

  12. Integrative Analyses of miRNA-mRNA Interactions Reveal let-7b, miR-128 and MAPK Pathway Involvement in Muscle Mass Loss in Sex-Linked Dwarf Chickens

    PubMed Central

    Luo, Wen; Lin, Shumao; Li, Guihuan; Nie, Qinghua; Zhang, Xiquan

    2016-01-01

    The sex-linked dwarf (SLD) chicken is an ideal model system for understanding growth hormone (GH)-action and growth hormone receptor (GHR) function because of its recessive mutation in the GHR gene. Skeletal muscle mass is reduced in the SLD chicken with a smaller muscle fiber diameter. Our previous study has presented the mRNA and miRNA expression profiles of the SLD chicken and normal chicken between embryo day 14 and seven weeks of age. However, the molecular mechanism of GHR-deficient induced muscle mass loss is still unclear, and the key molecules and pathways underlying the GHR-deficient induced muscle mass loss also remain to be illustrated. Here, by functional network analysis of the differentially expressed miRNAs and mRNAs between the SLD and normal chickens, we revealed that let-7b, miR-128 and the MAPK pathway might play key roles in the GHR-deficient induced muscle mass loss, and that the reduced cell division and growth are potential cellular processes during the SLD chicken skeletal muscle development. Additionally, we also found some genes and miRNAs involved in chicken skeletal muscle development, through the MAPK, PI3K-Akt, Wnt and Insulin signaling pathways. This study provides new insights into the molecular mechanism underlying muscle mass loss in the SLD chickens, and some regulatory networks that are crucial for chicken skeletal muscle development. PMID:26927061

  13. CD28/CTLA-4 ligands: the gene encoding CD86 (B70/B7.2) maps to the same region as CD80 (B7/B7.1) gene in human chromosome 3q13-q23.

    PubMed

    Fernández-Ruiz, E; Somoza, C; Sánchez-Madrid, F; Lanier, L L

    1995-05-01

    CD86 (B70/B7.2) is an antigen of the immunoglobulin superfamily expressed on monocytes, dendritic cells and activated B, T, and natural killer cells. CD86 was recently identified as a second ligand for the T cell antigens CD28 and CTLA-4, and plays an important role in the co-stimulation of T cells in a primary immune response. We report here the assignment of the CD86 gene to human chromosome 3 using Southern blot analysis on a panel of hamster x human somatic cell hybrid genomic DNA. Fluorescence hybridization in situ on metaphase chromosomes coupled with GTG banding (G-bands by trypsin using Giemsa staining) confirmed this assignment and localized the CD86 gene to 3q13-q23 region. The CD86 gene is, therefore, located in the proximity of the CD80 (B7/B7.1) gene, the first identified ligand for CD28 and CTLA-4, previously mapped to chromosome 3q13.3-q21. Deletions, inversions and insertions of chromosome 3q21-q26, as well as translocations of 3q21 with other chromosomes have been described in many cases of acute myeloid leukemia (AML), acute non-lymphocytic leukemia (ANLL), chronic myeloid leukemia (CML) and myelodisplastic syndromes (MDS), suggesting that this region contains several genes involved in the leukemic process.

  14. 40 CFR Table 6 to Subpart Bbbb of... - Model Rule-Requirements for Validating Continuous Emission Monitoring Systems (CEMS)

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    .... Nitrogen Oxides (Class I units only) a Method 7, 7A, 7B,7C, 7D, or 7E Method 3 or 3A. 2. Sulfur Dioxide... combustion plants with an aggregate plant combustion capacity greater than 250 tons per day of...

  15. 40 CFR Table 3 of Subpart Aaaa of... - Requirements for Validating Continuous Emission Monitoring Systems (CEMS)

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    .... Nitrogen Oxides (Class I units only) a Method 7, 7A, 7B, 7C, 7D, or 7E Method 3 or 3A. 2. Sulfur Dioxide... combustion plants with an aggregate plant combustion capacity more than 250 tons per day of municipal...

  16. 40 CFR Table 6 to Subpart Bbbb of... - Model Rule-Requirements for Validating Continuous Emission Monitoring Systems (CEMS)

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    .... Nitrogen Oxides (Class I units only) a Method 7, 7A, 7B,7C, 7D, or 7E Method 3 or 3A. 2. Sulfur Dioxide... combustion plants with an aggregate plant combustion capacity greater than 250 tons per day of...

  17. 40 CFR Table 6 to Subpart Bbbb of... - Model Rule-Requirements for Validating Continuous Emission Monitoring Systems (CEMS)

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    .... Nitrogen Oxides (Class I units only) a Method 7, 7A, 7B,7C, 7D, or 7E Method 3 or 3A. 2. Sulfur Dioxide... combustion plants with an aggregate plant combustion capacity greater than 250 tons per day of...

  18. 40 CFR Table 6 to Subpart Bbbb of... - Model Rule-Requirements for Validating Continuous Emission Monitoring Systems (CEMS)

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    .... Nitrogen Oxides (Class I units only) a Method 7, 7A, 7B,7C, 7D, or 7E Method 3 or 3A. 2. Sulfur Dioxide... combustion plants with an aggregate plant combustion capacity greater than 250 tons per day of...

  19. 40 CFR Table 6 to Subpart Bbbb of... - Model Rule-Requirements for Validating Continuous Emission Monitoring Systems (CEMS)

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    .... Nitrogen Oxides (Class I units only) a Method 7, 7A, 7B,7C, 7D, or 7E Method 3 or 3A. 2. Sulfur Dioxide... combustion plants with an aggregate plant combustion capacity greater than 250 tons per day of...

  20. 40 CFR Table 3 of Subpart Aaaa of... - Requirements for Validating Continuous Emission Monitoring Systems (CEMS)

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    .... Nitrogen Oxides (Class I units only) a Method 7, 7A, 7B, 7C, 7D, or 7E Method 3 or 3A. 2. Sulfur Dioxide... combustion plants with an aggregate plant combustion capacity more than 250 tons per day of municipal...

  1. Projection map of aquaporin-9 at 7 A resolution.

    PubMed

    Viadiu, Hector; Gonen, Tamir; Walz, Thomas

    2007-03-16

    Aquaporin-9, an aquaglyceroporin present in diverse tissues, is unique among aquaporins because it is not only permeable to water, urea and glycerol, but also allows passage of larger uncharged solutes. Single particle analysis of negatively stained recombinant rat aquaporin-9 revealed a particle size characteristic of the tetrameric organization of all members of the aquaporin family. Reconstitution of aquaporin-9 into two-dimensional crystals enabled us to calculate a projection map at 7 A resolution. The projection structure indicates a tetrameric structure, similar to GlpF, with each square-like monomer forming a pore. A comparison of the pore-lining residues between the crystal structure of GlpF and a homology model of aquaporin-9 locates substitutions in these residues predominantly to the hydrophobic edge of the tripathic pore of GlpF, providing first insights into the structural basis for the broader substrate specificity of aquaporin-9. PMID:17239399

  2. FalconSAT-7: a membrane space telescope

    NASA Astrophysics Data System (ADS)

    Andersen, Geoff; Asmolova, Olha; Dickinson, Thomas

    2014-06-01

    The USAF Academy Department of Physics is building FalconSAT-7, a membrane solar telescope to be deployed from a 3U CubeSat in LEO. The primary optic is a 0.2m photon sieve.—a diffractive element consisting of billions of tiny holes in an otherwise opaque polymer sheet. The membrane, its support structure, secondary optics, two imaging cameras and associated control/recording electronics are all packaged within half the CubeSat volume. Once in space the supporting pantograph structure is deployed, pulling the membrane flat under tension. The telescope will then be steered towards the Sun to gather images at H-alpha for transmission to the ground. Due for launch in 2016, FalconSAT-7 will serve as a pathfinder for future surveillance missions.

  3. FalconSAT-7: a membrane space solar telescope

    NASA Astrophysics Data System (ADS)

    Andersen, Geoff; Asmolova, Olha; McHarg, Matthew G.; Quiller, Trey; Maldonado, Carlos

    2016-07-01

    The US Air Force Academy of Physics has built FalconSAT-7, a membrane solar telescope to be deployed from a 3U CubeSat in LEO. The primary optic is a 0.2m photon sieve - a diffractive element consisting of billions of tiny circular dimples etched into a Kapton sheet. The membrane its support structure, secondary optics, two imaging cameras and associated control, recording electronics are packaged within half the CubeSat volume. Once in space the supporting pantograph structure is deployed, extending out and pulling the membrane flat under tension. The telescope will then be directed at the Sun to gather images at H-alpha for transmission to the ground. We will present details of the optical configuration, operation and performance of the flight telescope which has been made ready for launch in early 2017.

  4. Protection against aflatoxin B1-induced cytotoxicity by expression of the cloned aflatoxin B1-aldehyde reductases rat AKR7A1 and human AKR7A3.

    PubMed

    Bodreddigari, Sridevi; Jones, Laundette Knight; Egner, Patricia A; Groopman, John D; Sutter, Carrie Hayes; Roebuck, Bill D; Guengerich, F Peter; Kensler, Thomas W; Sutter, Thomas R

    2008-05-01

    The reduction of the aflatoxin B 1 (AFB 1) dialdehyde metabolite to its corresponding mono and dialcohols, catalyzed by aflatoxin B 1-aldehyde reductase (AFAR, rat AKR7A1, and human AKR7A3), is greatly increased in livers of rats treated with numerous chemoprotective agents. Recombinant human AKR7A3 has been shown to reduce the AFB 1-dialdehyde at rates greater than those of the rat AKR7A1. The activity of AKR7A1 or AKR7A3 may detoxify the AFB 1-dialdehyde, which reacts with proteins, and thereby inhibits AFB 1-induced toxicity; however, direct experimental evidence of this hypothesis was lacking. Two human B lymphoblastoid cell lines, designated pMF6/1A2/AKR7A1 and pMF6/1A2, were genetically engineered to stably express AKR7A1 and/or cytochrome P4501A2 (1A2). The pMF6/1A2/AKR7A1 cells were refractory to the cytotoxic effects of 3 ng/mL AFB 1, in comparison to pM6/1A2 cells, which were more sensitive. Diminished protection occurred at higher concentrations of AFB 1 in pMF6/1A2/AKR7A1 cells, suggesting that additional factors were influencing cell survival. COS-7 cells were transfected with either vector control, rat AKR7A1, or human AKR7A3, and the cells were treated with AFB 1-dialdehyde. There was a 6-fold increase in the dialdehyde LC 50, from 66 microM in vector-transfected cells to 400 microM in AKR7A1-transfected cells, and an 8.5-fold increase from 35 microM in vector-transfected cells to 300 microM in AKR7A3-transfected cells. In both cases, this protective effect of the AFAR enzyme was accompanied by a marked decrease in protein adducts. Fractionation of the cellular protein showed that the mitochondria/nuclei and microsomal fractions contained the highest concentration of protein adducts. The levels of human AKR7A3 and AKR7A2 were measured in 12 human liver samples. The expression of AKR7A3 was detectable in all livers and lower than those of AKR7A2 in 11 of the 12 samples. Overall, these results provide the first direct evidence of a role for rat AKR7A1

  5. Feige 7 - A hot, rotating magnetic white dwarf

    NASA Technical Reports Server (NTRS)

    Liebert, J.; Angel, J. R. P.; Stockman, H. S.; Spinrad, H.; Beaver, E. A.

    1977-01-01

    Results are reported for image-tube-scanner and digicon observations of Feige 7, a faint blue star identified as a probable white dwarf. It is found that this star is a magnetic white dwarf showing a very rich spectrum with Zeeman subcomponents of both hydrogen and neutral helium as well as periodic spectrum and circular-polarization variations. A polarization period of 2.2 hr is computed, and a surface magnetic-field strength of about 18 MG is determined by matching features of the absorption spectrum to Zeeman components. It is suggested that the only reasonable explanation for the periodic variations in circular polarization is an oblique rotator with the spin axis approximately in the plane of the sky and tilted by about 24 deg to the magnetic axis. An effective temperature in the range from 20,000 to 25,000 K is estimated, an absolute magnitude of about 10.5 is derived, and the atmosphere is shown to be helium-dominated. The evolution of Feige 7 is discussed in terms of possible magnetic-field effects on atmospheric composition, rotation velocity (5.5 km/s for a radius of 7000 km), and the origin of white-dwarf magnetic fields.

  6. Sol-gel synthesis of Na4Fe3(PO4)2(P2O7)/C nanocomposite for sodium ion batteries and new insights into microstructural evolution during sodium extraction

    NASA Astrophysics Data System (ADS)

    Wu, Xuehang; Zhong, Guiming; Tang, Zheng; Yang, Yong

    2016-09-01

    A mixed polyanionic Na4Fe3(PO4)2(P2O7)/C nanocomposite is synthesized via a sol-gel route. The phosphate raw material is transformed to the mixed phosphate-pyrophosphate with high phase purity via a self-condensation reaction at 500 °C. Na4Fe3(PO4)2(P2O7)/C can deliver an initial capacity of 110 mAh g-1 at 0.05C with the average discharge voltage approaching 3.1 V. The nanocomposite shows excellent rate capability because of the presence of an in-situ formed 3-D network of carbon. At 10 C rate, the nanocomposite delivers a discharge capacity of 78 mAh g-1 at 25 mAh °C and 82 mAh g-1 at 55 °C. The nanocomposite has a good long-term cycling stability, retaining 89% of the initial discharge capacity after 300 cycles. In-situ XRD results demonstrate that the sodium insertion/extraction process in Na4Fe3(PO4)2(P2O7) is an imperfect solid-solution reaction with an obvious local lattice distortion instead of an ideal solid-solution reaction. Using a solid-state NMR technique, it is further found that the sodium extraction from the Na1, Na3, and Na4 sites causes an obvious change in local structure. However, the local structure of Na2 remains unchanged, which may aid the stability of the host structure.

  7. Predictive Value of BRCA1, ERCC1, ATP7B, PKM2, TOPOI, TOPΟ-IIA, TOPOIIB and C-MYC Genes in Patients with Small Cell Lung Cancer (SCLC) Who Received First Line Therapy with Cisplatin and Etoposide

    PubMed Central

    Karachaliou, Niki; Papadaki, Chara; Lagoudaki, Eleni; Trypaki, Maria; Sfakianaki, Maria; Koutsopoulos, Anastasios; Mavroudis, Dimitris; Stathopoulos, Efstathios; Georgoulias, Vassilis; Souglakos, John

    2013-01-01

    Background The aim of the study was to evaluate the predictive value of genes involved in the action of cisplatin-etoposide in Small Cell Lung Cancer (SCLC). Methods 184 SCLC patients’ primary tumour samples were analyzed for ERCCI, BRCA1, ATP7B, PKM2 TOPOI, TOPOIIA, TOPOIIB and C-MYC mRNA expression. All patients were treated with cisplatin-etoposide. Results The patients’ median age was 63 years and 120 (65%) had extended stage, 75 (41%) had increased LDH serum levels and 131 (71%) an ECOG performance status was 0-1. Patients with limited stage, whose tumours expressed high ERCC1 (p=0.028), PKM2 (p=0.046), TOPOI (p=0.008), TOPOIIA (p=0.002) and TOPOIIB (p<0.001) mRNA had a shorter Progression Free Survival (PFS). In limited stage patients, high expression of ERCC1 (p=0.014), PKM2 (p=0.026), TOPOIIA (p=0.021) and TOPOIIB (p=0.019) was correlated with decreased median overall survival (mOS) while in patients with extended stage, only high TOPOIIB expression had a negative impact on Os (p=0.035). The favorable expression signature expression signature (low expression of ERCC1, PKM2, TOPOIIA and TOPOIIB) was correlated with significantly better PFS and Os in both LS-SCLC (p<0.001 and p=0.007, respectively) and ES-SCLC (p=0.007 and (p=0.011, respectively) group. The unfavorable expression signature was an independent predictor for poor PFS (HR: 3.18; p=0.002 and HR: 3.14; p=0.021) and Os (HR: 4.35; p=0.001and HR: 3.32; p=0.019) in both limited and extended stage, respectively. Conclusions Single gene’s expression analysis as well as the integrated analysis of ERCC1, PKM2, TOPOIIA and TOPOIIB may predict treatment outcome in patients with SCLC. These findings should be further validated in a prospective study. PMID:24058603

  8. Test plan for the M-100 container, (model M-101/7A/12/90) docket 96-43-7A, type A container. Revision 1

    SciTech Connect

    Kelly, D.L.

    1997-07-22

    This report concerns the packaging configurations being tested by the U.S. DOE and its contractors, and according to U.S. DOT specification 7A Type A (DOT-7A) requirements. The objective of this Test Plan is to describe the testing for the qualification of the M-100 Container, Model M-101/7A/12/90 as a DOT-7A Type A packaging. This packaging system is designed to ship Type A solid radioactive materials, normal form, Form Number 1, Form Number 2, and Form Number 3.

  9. The effect of CYP7A1 polymorphisms on lipid responses to fenofibrate

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: CYP7A1 encodes cholesterol 7a-hydroxylase, an enzyme crucial to cholesterol homeostasis. Its transcriptional activity is downregulated by fenofibrate. The goal of this study s to determine the effect of CYP7A1 polymorphisms on lipid changes in response to fenofibrate. Methods: We exam...

  10. 17 CFR 260.7a-24 - Words relating to periods of time in the past.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 17 Commodity and Securities Exchanges 3 2010-04-01 2010-04-01 false Words relating to periods of time in the past. 260.7a-24 Section 260.7a-24 Commodity and Securities Exchanges SECURITIES AND... Requirements As to Contents § 260.7a-24 Words relating to periods of time in the past. Unless the...

  11. 17 CFR 260.7a-25 - Words relating to the future.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 17 Commodity and Securities Exchanges 3 2010-04-01 2010-04-01 false Words relating to the future. 260.7a-25 Section 260.7a-25 Commodity and Securities Exchanges SECURITIES AND EXCHANGE COMMISSION... § 260.7a-25 Words relating to the future. Unless the context clearly shows otherwise, whenever...

  12. Pri-let-7a-1 rs10739971 polymorphism is associated with gastric cancer prognosis and might affect mature let-7a expression

    PubMed Central

    Li, Ying; Xu, Qian; Liu, Jingwei; He, Caiyun; Yuan, Quan; Xing, Chengzhong; Yuan, Yuan

    2016-01-01

    The relationship between the pri-let-7a-1 rs10739971 polymorphism and gastric cancer (GC) risk has been reported. However, the role of this polymorphism in the prognosis of GC remains largely elusive. Sequenom MassARRAY platform method and the polymerase chain reaction (PCR)-restriction fragment length polymorphism were used to investigate pri-let-7a-1 rs10739971 G→A in 334 GC patients. Real-time PCR detected expression of mature let-7a in serum and tissue. Patients with AA or GA+AA genotypes of the pri-let-7a-1 rs10739971 polymorphism demonstrated significantly longer survival time than those with the wild GG genotype. Stratified analysis indicated that survival time was significantly longer in women with AA or GA+AA genotypes and in Borrmann type I/II patients with GA heterozygote or GA+AA genotypes. AA genotype was more frequent in the lymphatic-metastasis-negative subgroup. Serum mature let-7a expression in healthy people with the GA heterozygote and the GA+AA genotype was higher than in those with the GG genotype, and the difference remained significant in the female healthy subgroup. Pri-let-7a-1 rs10739971 polymorphism might be a biomarker for GC prognosis, especially for female and Borrmann type I/II patients. The pri-let-7a-1 rs10739971 polymorphism might affect serum mature let-7a expression, and partly explain the mechanism of the relationship between the pri-let-7a-1 rs10739971 polymorphism and GC survival. PMID:27445488

  13. New lithium copper borates with BO3 triangles: Li6CuB4O10, Li3CuB3O7, Li8Cu7B14O32, and Li2Cu9B12O28.

    PubMed

    Kuratieva, N V; Bànki, M; Tsirlin, A A; Eckert, J; Ehrenberg, H; Mikhailova, D

    2013-12-16

    Crystal structures of three new lithium copper borates, Li3CuB3O7, Li8Cu7B14O32, and Li2Cu9B12O28, and a new Li6CuB4O10 polymorph were solved by single-crystal X-ray diffraction. In all of the structures, the boron cations form BO3 triangles, which are connected with each other and with copper polyhedra only via corners in Li6CuB4O10 and Li3CuB3O7 and via both corners and edges in Li8Cu7B14O32 and Li2Cu9B12O28. The Li3CuB3O7 and Li8Cu7B14O32 compounds were synthesized as pure samples with only trace amounts of impurities; hence, their magnetic properties could be investigated and analyzed in terms of underlying magnetic couplings. Other compositions always represented multiphase mixtures. Li3CuB3O7 features infinite Cu,O chains formed by Cu2O6 units consisting of edge-shared CuO4 squares. Together with two apical oxygen atoms with long interatomic Cu-O distances of 2.7-2.8 Å, the Cu2O6 units form chains extended along the a axis. These pseudochains are responsible for strong anisotropic thermal expansion behavior. The temperature dependence of the magnetization between 4 and 380 K for Li3CuB3O7 could be fit well by a spin-dimer model. The magnetic susceptibility of Li8Cu7B14O32 showed a more complex temperature dependence, with two different Curie-Weiss regimes in the temperature range of 2-380 K. PMID:24266792

  14. Proteomic analysis reveals that COP9 signalosome complex subunit 7A (CSN7A) is essential for the phase transition of migratory locust.

    PubMed

    Tong, Xi-Wen; Chen, Bing; Huang, Li-Hua; Feng, Qi-Li; Kang, Le

    2015-01-01

    The migratory locust displays a reversible, density-dependent transition between the two phases of gregaria and solitaria. This phenomenon is a typical kind of behavior plasticity. Here, we report that COP9 signalosome complex subunit 7A (CSN7A) is involved in the regulation of locust phase transition. Firstly, 90 proteins were identified to express differentially between the two phases by quantitative proteomic analysis. Gregaria revealed higher levels in proteins related to structure formation, melanism and energy metabolism, whereas solitaria had more abundant proteins related to digestion, absorption and chemical sensing. Subsequently, ten proteins including CSN7A were found to reveal differential mRNA expression profiles between the two phases. The CSN7A had higher mRNA level in the gregaria as compared with the solitaria, and the mRNA amount in the gregaria decreased remarkably during the 32 h-isolation. However, the mRNA level in the solitaria kept constant during the crowding rearing. Finally and importantly, RNA interference of CSN7A in gregaria resulted in obvious phase transition towards solitaria within 24 h. It suggests that CSN7A plays an essential role in the transition of gregaria towards solitaria in the migratory locust. To our knowledge, it's the first time to report the role of CSN in behavior plasticity of animals. PMID:26212173

  15. Proteomic analysis reveals that COP9 signalosome complex subunit 7A (CSN7A) is essential for the phase transition of migratory locust

    PubMed Central

    Tong, Xi-Wen; Chen, Bing; Huang, Li-Hua; Feng, Qi-Li; Kang, Le

    2015-01-01

    The migratory locust displays a reversible, density-dependent transition between the two phases of gregaria and solitaria. This phenomenon is a typical kind of behavior plasticity. Here, we report that COP9 signalosome complex subunit 7A (CSN7A) is involved in the regulation of locust phase transition. Firstly, 90 proteins were identified to express differentially between the two phases by quantitative proteomic analysis. Gregaria revealed higher levels in proteins related to structure formation, melanism and energy metabolism, whereas solitaria had more abundant proteins related to digestion, absorption and chemical sensing. Subsequently, ten proteins including CSN7A were found to reveal differential mRNA expression profiles between the two phases. The CSN7A had higher mRNA level in the gregaria as compared with the solitaria, and the mRNA amount in the gregaria decreased remarkably during the 32 h-isolation. However, the mRNA level in the solitaria kept constant during the crowding rearing. Finally and importantly, RNA interference of CSN7A in gregaria resulted in obvious phase transition towards solitaria within 24 h. It suggests that CSN7A plays an essential role in the transition of gregaria towards solitaria in the migratory locust. To our knowledge, it’s the first time to report the role of CSN in behavior plasticity of animals. PMID:26212173

  16. Electrosynthesis of Rh2(dpf)4(R) where dpf = N,N'-diphenylformamidinate anion and R = CH3, C2H5, C3H7, C4H9 or C5H11.

    PubMed

    Bear, J L; Van Caemelbecke, E; Ngubane, S; Da-Riz, V; Kadish, K M

    2011-03-21

    The electrosynthesis of Rh(2)(dpf)(4)(R) where dpf is the N,N'-diphenylformamidinate anion and R = CH(3), C(2)H(5), C(3)H(7), C(4)H(9) or C(5)H(11) was carried out in THF containing 0.2 M tetra-n-butylammonium perchlorate (TBAP) and one of several alkyl iodides represented as RI. The initial step in the reaction involved a one-electron reduction of the Rh(2)(4+) unit in Rh(2)(dpf)(4) to its Rh(2)(3+) form followed by a homogeneous reaction involving electrogenerated [Rh(2)(dpf)(4)](-) and the alkyl iodide in solution to give Rh(2)(dpf)(4)(R). The homogeneously generated Rh(2)(5+) product was then immediately reduced by a second electron at the potential where [Rh(2)(dpf)(4)(R)](-) is generated, giving [Rh(2)(dpf)(4)(R)](-) which contains a Rh(2)(4+) center as a final product of an electrochemical ECE mechanism. The electrosynthesized [Rh(2)(dpf)(4)(CH(3))](-) derivative could be reoxidized to Rh(2)(dpf)(4)(CH(3)) on the reverse potential sweep and both forms of the CH(3) bonded derivative were in situ characterized by cyclic voltammetry combined with UV-visible and/or ESR spectroscopy. The reversible Rh(2)(4+/3+) process of Rh(2)(dpf)(4) is located at E(1/2) = -1.11 V in THF, 0.2 M TBAP while the electrogenerated Rh(2)(dpf)(4)(R) products are substantially easier to reduce, with E(p) values for the Rh(2)(5+/4+) couples ranging from -0.50 to -0.54 V vs. SCE depending upon the specific R group.

  17. The new high-pressure borate Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O-Formation of edge-sharing BO{sub 4} tetrahedra in a hydrated borate

    SciTech Connect

    Neumair, Stephanie C.; Kaindl, Reinhard; Huppertz, Hubert

    2012-01-15

    The new borate hydrate Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O was synthesized under high-pressure/high-temperature conditions of 6 GPa and 880 Degree-Sign C in a Walker-type multianvil apparatus. The compound crystallizes in the orthorhombic space group Pbam (Z=2) with the lattice parameters a=819.0(2), b=2016.9(4), c=769.9(2) pm, V=1.2717(4) nm{sup 3}, R{sub 1}=0.0758, wR{sub 2}=0.0836 (all data). The new structure type of Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O is built up from corner-sharing BO{sub 4} tetrahedra forming corrugated layers, that are interconnected among each other by two edge-sharing BO{sub 4} tetrahedra (B{sub 2}O{sub 6} units) forming Z-shaped channels. Interestingly, the here presented structure of Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O is closely related to the structures of M{sub 6}B{sub 22}O{sub 39}{center_dot}H{sub 2}O (M=Fe, Co), which exhibit BO{sub 4} tetrahedra in an intermediate state on the way to edge-sharing BO{sub 4} tetrahedra. - Graphical Abstract: The new high-pressure borate hydrate Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O is built up from corner-sharing BO{sub 4} tetrahedra forming corrugated layers, that are interconnected among each other by two edge-sharing BO{sub 4} tetrahedra (B{sub 2}O{sub 6} units). In this paper we report on synthesis, structural details, and properties of the new compound Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O. Highlights: Black-Right-Pointing-Pointer High-pressure/high-temperature synthesis of the new borate hydrate Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O. Black-Right-Pointing-Pointer In the structure of Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O, two B{sub 2}O{sub 6} units are connected to 'vierer' rings. Black-Right-Pointing-Pointer Pressure favours the formation of edge-sharing BO{sub 4} tetrahedra in the chemistry of borates.

  18. THSD7A staining of membranous glomerulopathy in clinical practice reveals cases with dual autoantibody positivity.

    PubMed

    Larsen, Christopher P; Cossey, L Nicholas; Beck, Laurence H

    2016-04-01

    Thrombospondin type I domain-containing 7A (THSD7A) is a known antigenic target of autoantibodies leading to primary membranous glomerulopathy and was reported to account for ~10% of phospholipase A2 receptor (PLA2R)-negative membranous glomerulopathy. It has been proposed that PLA2R and THSD7A autoantibodies are mutually exclusive in membranous glomerulopathy. We validated an immunohistochemical assay to investigate for THSD7A-associated membranous glomerulopathy and utilized it in 258 consecutive native kidney biopsies, which showed membranous glomerulopathy in our laboratory, with the exception of membranous lupus nephritis. Membranous glomerulopathy stained positive for THSD7A-only in 7 (3%) cases, PLA2R-only in 141 (55%) cases, and showed dual positivity for THSD7A and PLA2R in 2 (1%) cases. Serologic testing for antibodies to PLA2R and THSD7A was performed in a subset of these patients. There was 100% correlation between positive THSD7A and/or PLA2R tissue staining and the presence of the corresponding autoantibodies in the serum including the two cases with dual positive THSD7A and PLA2R antibodies. We describe and provide a protocol for detection of THSD7A-associated membranous glomerulopathy in clinical practice. The cases with dual THSD7A and PLA2R positivity show that these autoantibodies are not mutually exclusive. They also emphasize the importance of using a panel-based approach when subtyping membranous glomerulopathy as a patient could conceptually be identified and treated based on anti-PLA2R titers, but still have anti-THSD7A antibodies driving persistent disease.

  19. THSD7A staining of membranous glomerulopathy in clinical practice reveals cases with dual autoantibody positivity

    PubMed Central

    Larsen, Christopher P; Cossey, L Nicholas; Beck, Laurence H

    2016-01-01

    Thrombospondin type I domain-containing 7A (THSD7A) is a known antigenic target of autoantibodies leading to primary membranous glomerulopathy and was reported to account for ~10% of phospholipase A2 receptor (PLA2R)-negative membranous glomerulopathy. It has been proposed that PLA2R and THSD7A autoantibodies are mutually exclusive in membranous glomerulopathy. We validated an immunohistochemical assay to investigate for THSD7A-associated membranous glomerulopathy and utilized it in 258 consecutive native kidney biopsies, which showed membranous glomerulopathy in our laboratory, with the exception of membranous lupus nephritis. Membranous glomerulopathy stained positive for THSD7A-only in 7 (3%) cases, PLA2R-only in 141 (55%) cases, and showed dual positivity for THSD7A and PLA2R in 2 (1%) cases. Serologic testing for antibodies to PLA2R and THSD7A was performed in a subset of these patients. There was 100% correlation between positive THSD7A and/or PLA2R tissue staining and the presence of the corresponding autoantibodies in the serum including the two cases with dual positive THSD7A and PLA2R antibodies. We describe and provide a protocol for detection of THSD7A-associated membranous glomerulopathy in clinical practice. The cases with dual THSD7A and PLA2R positivity show that these autoantibodies are not mutually exclusive. They also emphasize the importance of using a panel-based approach when subtyping membranous glomerulopathy as a patient could conceptually be identified and treated based on anti-PLA2R titers, but still have anti-THSD7A antibodies driving persistent disease. PMID:26847174

  20. ANALYSIS OF DWPF SLUDGE BATCH 7A (MACROBATCH 8) POUR STREAM SAMPLES

    SciTech Connect

    Johnson, F.

    2012-05-01

    The Defense Waste Processing Facility (DWPF) began processing Sludge Batch 7a (SB7a), also referred to as Macrobatch 8 (MB8), in June 2011. SB7a is a blend of the heel of Tank 40 from Sludge Batch 6 (SB6) and the SB7a material that was transferred to Tank 40 from Tank 51. SB7a was processed using Frit 418. During processing of each sludge batch, the DWPF is required to take at least one glass sample to meet the objectives of the Glass Product Control Program (GPCP), which is governed by the DWPF Waste Compliance Plan, and to complete the necessary Production Records so that the final glass product may be disposed of at a Federal Repository. Three pour stream glass samples and two Melter Feed Tank (MFT) slurry samples were collected while processing SB7a. These additional samples were taken during SB7a to understand the impact of antifoam and the melter bubblers on glass redox chemistry. The samples were transferred to the Savannah River National Laboratory (SRNL) where they were analyzed. The following conclusions were drawn from the analytical results provided in this report: (1) The sum of oxides for the official SB7a pour stream glass is within the Product Composition Control System (PCCS) limits (95-105 wt%). (2) The average calculated Waste Dilution Factor (WDF) for SB7a is 2.3. In general, the measured radionuclide content of the official SB7a pour stream glass is in good agreement with the calculated values from the Tank 40 dried sludge results from the SB7a Waste Acceptance Program Specification (WAPS) sample. (3) As in previous pour stream samples, ruthenium and rhodium inclusions were detected by Scanning Electron Microscopy-Electron Dispersive Spectroscopy (SEM-EDS) in the official SB7a pour stream sample. (4) The Product Consistency Test (PCT) results indicate that the official SB7a pour stream glass meets the waste acceptance criteria for durability with a normalized boron release of 0.64 g/L, which is an order of magnitude less than the Environmental

  1. Autoantibodies against thrombospondin type 1 domain–containing 7A induce membranous nephropathy

    PubMed Central

    Tomas, Nicola M.; Hoxha, Elion; Reinicke, Anna T.; Fester, Lars; Helmchen, Udo; Gerth, Jens; Bachmann, Friederike; Budde, Klemens; Zahner, Gunther; Rune, Gabriele; Lambeau, Gerard; Meyer-Schwesinger, Catherine

    2016-01-01

    Membranous nephropathy (MN) is the most common cause of nephrotic syndrome in adults, and one-third of patients develop end-stage renal disease (ESRD). Circulating autoantibodies against the podocyte surface antigens phospholipase A2 receptor 1 (PLA2R1) and the recently identified thrombospondin type 1 domain–containing 7A (THSD7A) are assumed to cause the disease in the majority of patients. The pathogenicity of these antibodies, however, has not been directly proven. Here, we have reported the analysis and characterization of a male patient with THSD7A-associated MN who progressed to ESRD and subsequently underwent renal transplantation. MN rapidly recurred after transplantation. Enhanced staining for THSD7A was observed in the kidney allograft, and detectable anti-THSD7A antibodies were present in the serum before and after transplantation, suggesting that these antibodies induced a recurrence of MN in the renal transplant. In contrast to PLA2R1, THSD7A was expressed on both human and murine podocytes, enabling the evaluation of whether anti-THSD7A antibodies cause MN in mice. We demonstrated that human anti-THSD7A antibodies specifically bind to murine THSD7A on podocyte foot processes, induce proteinuria, and initiate a histopathological pattern that is typical of MN. Furthermore, anti-THSD7A antibodies induced marked cytoskeletal rearrangement in primary murine glomerular epithelial cells as well as in human embryonic kidney 293 cells. Our findings support a causative role of anti-THSD7A antibodies in the development of MN. PMID:27214550

  2. Autoantibodies against thrombospondin type 1 domain-containing 7A induce membranous nephropathy.

    PubMed

    Tomas, Nicola M; Hoxha, Elion; Reinicke, Anna T; Fester, Lars; Helmchen, Udo; Gerth, Jens; Bachmann, Friederike; Budde, Klemens; Koch-Nolte, Friedrich; Zahner, Gunther; Rune, Gabriele; Lambeau, Gerard; Meyer-Schwesinger, Catherine; Stahl, Rolf A K

    2016-07-01

    Membranous nephropathy (MN) is the most common cause of nephrotic syndrome in adults, and one-third of patients develop end-stage renal disease (ESRD). Circulating autoantibodies against the podocyte surface antigens phospholipase A2 receptor 1 (PLA2R1) and the recently identified thrombospondin type 1 domain-containing 7A (THSD7A) are assumed to cause the disease in the majority of patients. The pathogenicity of these antibodies, however, has not been directly proven. Here, we have reported the analysis and characterization of a male patient with THSD7A-associated MN who progressed to ESRD and subsequently underwent renal transplantation. MN rapidly recurred after transplantation. Enhanced staining for THSD7A was observed in the kidney allograft, and detectable anti-THSD7A antibodies were present in the serum before and after transplantation, suggesting that these antibodies induced a recurrence of MN in the renal transplant. In contrast to PLA2R1, THSD7A was expressed on both human and murine podocytes, enabling the evaluation of whether anti-THSD7A antibodies cause MN in mice. We demonstrated that human anti-THSD7A antibodies specifically bind to murine THSD7A on podocyte foot processes, induce proteinuria, and initiate a histopathological pattern that is typical of MN. Furthermore, anti-THSD7A antibodies induced marked cytoskeletal rearrangement in primary murine glomerular epithelial cells as well as in human embryonic kidney 293 cells. Our findings support a causative role of anti-THSD7A antibodies in the development of MN.

  3. 49 CFR 178.350 - Specification 7A; general packaging, Type A.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 2 2010-10-01 2010-10-01 false Specification 7A; general packaging, Type A. 178... FOR PACKAGINGS Specifications for Packagings for Class 7 (Radioactive) Materials § 178.350 Specification 7A; general packaging, Type A. (a) Each packaging must meet all applicable requirements of...

  4. 76 FR 4349 - Revised Jurisdictional Thresholds for Section 7a of The Clayton Act

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-25

    .... Section 7A of the Clayton Act, 15 U.S.C. 18a, as added by the Hart-Scott-Rodino ] Antitrust Improvements.... ACTION: Notice. SUMMARY: The Federal Trade Commission announces the revised thresholds for the Hart-Scott..., Bureau of Competition, Premerger Notification Office, (202) 326-3100. Authority: 16 U.S.C. 7A....

  5. 17 CFR 260.7a-17 - Quality, color and size of paper.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... paper. 260.7a-17 Section 260.7a-17 Commodity and Securities Exchanges SECURITIES AND EXCHANGE COMMISSION... Quality, color and size of paper. The application, statement or report, including all amendments and, where practicable, all papers and documents filed as a part thereof, shall be on good quality,...

  6. Let7a involves in neural stem cell differentiation relating with TLX level

    SciTech Connect

    Song, Juhyun; Cho, Kyoung Joo; Oh, Yumi; Lee, Jong Eun

    2015-07-10

    Neural stem cells (NSCs) have the potential for differentiation into neurons known as a groundbreaking therapeutic solution for central nervous system (CNS) diseases. To resolve the therapeutic efficiency of NSCs, recent researchers have focused on the study on microRNA's role in CNS. Some micro RNAs have been reported significant functions in NSC self-renewal and differentiation through the post-transcriptional regulation of neurogenesis genes. MicroRNA-Let7a (Let7a) has known as the regulator of diverse cellular mechanisms including cell differentiation and proliferation. In present study, we investigated whether Let7a regulates NSC differentiation by targeting the nuclear receptor TLX, which is an essential regulator of NSC self-renewal, proliferation and differentiation. We performed the following experiments: western blot analysis, TaqMan assay, RT-PCR, and immunocytochemistry to confirm the alteration of NSCs. Our data showed that let7a play important roles in controlling NSC fate determination. Thus, manipulating Let-7A and TLX could be a novel strategy to enhance the efficiency of NSC's neuronal differentiation for CNS disorders. - Highlights: • Let7a influences on NSC differentiation and proliferation. • Let7a involves in mainly NSC differentiation rather than proliferation. • Let7a positively regulates the TLX expression.

  7. Zebrafish Thsd7a is a neural protein required for angiogenic patterning during development.

    PubMed

    Wang, Chieh-Huei; Chen, I-Hui; Kuo, Meng-Wei; Su, Pei-Tsu; Lai, Zih-Yin; Wang, Chian-Huei; Huang, Wei-Chang; Hoffman, Jana; Kuo, Calvin J; You, May-Su; Chuang, Yung-Jen

    2011-06-01

    Angiogenesis is a highly organized process under the control of guidance cues that direct endothelial cell (EC) migration. Recently, many molecules that were initially described as regulators of neural guidance were subsequently shown to also direct EC migration. Here, we report a novel protein, thrombospondin type I domain containing 7A (Thsd7a), that is a neural molecule required for directed EC migration during embryonic angiogenesis in zebrafish. Thsd7a is a vertebrate conserved protein. Zebrafish thsd7a transcript was detected along the ventral edge of the neural tube in the developing zebrafish embryos, correlating with the growth path of angiogenic intersegmental vessels (ISVs). Morpholino-knockdown of Thsd7a caused a lateral deviation of angiogenic ECs below the thsd7a-expressing sites, resulting in aberrant ISV patterning. Collectively, our study shows that zebrafish Thsd7a is a neural protein required for ISV angiogenesis, and suggests an important role of Thsd7a in the neurovascular interaction during zebrafish development.

  8. ZBTB7A acts as a tumor suppressor through the transcriptional repression of glycolysis

    PubMed Central

    Liu, Xue-Song; Haines, Jenna E.; Mehanna, Elie K.; Genet, Matthew D.; Ben-Sahra, Issam; Asara, John M.; Manning, Brendan D.

    2014-01-01

    Elevated glycolysis is a common metabolic trait of cancer, but what drives such metabolic reprogramming remains incompletely clear. We report here a novel transcriptional repressor-mediated negative regulation of glycolysis. ZBTB7A, a member of the POK (POZ/BTB and Krüppel) transcription repressor family, directly binds to the promoter and represses the transcription of critical glycolytic genes, including GLUT3, PFKP, and PKM. Analysis of The Cancer Genome Atlas (TCGA) data sets reveals that the ZBTB7A locus is frequently deleted in many human tumors. Significantly, reduced ZBTB7A expression correlates with up-regulation of the glycolytic genes and poor survival in colon cancer patients. Remarkably, while ZBTB7A-deficient tumors progress exceedingly fast, they exhibit an unusually heightened sensitivity to glycolysis inhibition. Our study uncovers a novel tumor suppressor role of ZBTB7A in directly suppressing glycolysis. PMID:25184678

  9. Vimentin phosphorylation and assembly are regulated by the small GTPase Rab7a.

    PubMed

    Cogli, Laura; Progida, Cinzia; Bramato, Roberta; Bucci, Cecilia

    2013-06-01

    Intermediate filaments are cytoskeletal elements important for cell architecture. Recently it has been discovered that intermediate filaments are highly dynamic and that they are fundamental for organelle positioning, transport and function thus being an important regulatory component of membrane traffic. We have identified, using the yeast two-hybrid system, vimentin, a class III intermediate filament protein, as a Rab7a interacting protein. Rab7a is a member of the Rab family of small GTPases and it controls vesicular membrane traffic to late endosomes and lysosomes. In addition, Rab7a is important for maturation of phagosomes and autophagic vacuoles. We confirmed the interaction in HeLa cells by co-immunoprecipitation and pull-down experiments, and established that the interaction is direct using bacterially expressed recombinant proteins. Immunofluorescence analysis on HeLa cells indicate that Rab7a-positive vesicles sometimes overlap with vimentin filaments. Overexpression of Rab7a causes an increase in vimentin phosphorylation at different sites and causes redistribution of vimentin in the soluble fraction. Consistently, Rab7a silencing causes an increase of vimentin present in the insoluble fraction (assembled). Also, expression of Charcot-Marie-Tooth 2B-causing Rab7a mutant proteins induces vimentin phosphorylation and increases the amount of vimentin in the soluble fraction. Thus, modulation of expression levels of Rab7a wt or expression of Rab7a mutant proteins changes the assembly of vimentin and its phosphorylation state indicating that Rab7a is important for the regulation of vimentin function.

  10. Computational and anti-tumor studies of 7a-Aza-B-homostigmast-5-eno [7a, 7-d] tetrazole-3β-yl chloride

    NASA Astrophysics Data System (ADS)

    Alam, Mahboob; Alam, Mohammad Jane; Nami, Shahab A. A.; Lee, Dong-Ung; Azam, Mohammad; Ahmad, Shabbir

    2016-03-01

    The present paper reports the detailed computational study including molecular docking of a biologically active steroidal tetrazole, 7a-Aza-B-homostigmast-5-eno [7a,7-d] tetrazole-3β-yl chloride. The molecular structure, IR and NMR (13C and 1H) spectra of the tetrazole were interpreted by comparing the experimental results with the theoretical, B3LYP/6-311G(d,p) calculations. The vibrational bands appearing in the FTIR are assigned with great accuracy using animated modes. Molecular properties like HOMO-LUMO analysis, chemical reactivity descriptors, MEP mapping, dipole moment and natural atomic charges have been presented at the same level of theory. The theoretical results are found in good correlation with the experimental data. Moreover, the Hirshfeld analysis was carried out to ascertain the secondary interactions and associated 2D fingerprint plots. The in vitro anti-tumor activity of 7a-Aza-B-homostigmast-5-eno [7a,7-d] tetrazole-3β-yl chloride has also been carried out against five human tumor cell lines. Doxorubicin is used as a standard drug for the in vitro anti-tumor screening.

  11. DFT, Hirshfeld surfaces, spectral and in vivo cytotoxic studies of 7a-Aza-B-homostigmast-5-eno [7a,7-d] tetrazole

    NASA Astrophysics Data System (ADS)

    Alam, Mahboob; Alam, Mohammad Jane; Nami, Shahab A. A.; Khan, Mohd Shoeb; Ahmad, Shabbir; Lee, Dong-Ung

    2015-11-01

    The DFT (B3LYP) calculations on a synthetic steroidal molecule 7a-Aza-B-homostigmast-5-eno [7a,7-d] tetrazole, C29H48N4, have been performed. The molecular structure, IR and NMR (13C and 1H) spectra of the present compound were interpreted using experiments (XRD, FTIR, NMR) as well as theoretical, B3LYP/6-311 + G(2d,p), calculations. The vibrational bands appearing in FTIR are assigned with great accuracy using animated modes. Molecular properties like HOMO-LUMO analysis, chemical reactivity descriptors, MEP mapping, dipole moment and Mullikan's atomic charges have been presented at the same level of theory. The theoretical results are found in good correlation with experimental data. Moreover, the Hirshfeld analysis was carried out to ascertain the secondary interactions and associated 2D fingerprint plots. The in vivo cytotoxicity of 7a-Aza-B-homostigmast-5-eno [7a,7-d] has also been carried out against brine shrimp nauplii by lethality bioassay.

  12. Computational and anti-tumor studies of 7a-Aza-B-homostigmast-5-eno [7a, 7-d] tetrazole-3β-yl chloride

    NASA Astrophysics Data System (ADS)

    Alam, Mahboob; Alam, Mohammad Jane; Nami, Shahab A. A.; Lee, Dong-Ung; Azam, Mohammad; Ahmad, Shabbir

    2016-03-01

    The present paper reports the detailed computational study including molecular docking of a biologically active steroidal tetrazole, 7a-Aza-B-homostigmast-5-eno [7a,7-d] tetrazole-3β-yl chloride. The molecular structure, IR and NMR (13C and 1H) spectra of the tetrazole were interpreted by comparing the experimental results with the theoretical, B3LYP/6-311G(d,p) calculations. The vibrational bands appearing in the FTIR are assigned with great accuracy using animated modes. Molecular properties like HOMO-LUMO analysis, chemical reactivity descriptors, MEP mapping, dipole moment and natural atomic charges have been presented at the same level of theory. The theoretical results are found in good correlation with the experimental data. Moreover, the Hirshfeld analysis was carried out to ascertain the secondary interactions and associated 2D fingerprint plots. The in vitro anti-tumor activity of 7a-Aza-B-homostigmast-5-eno [7a,7-d] tetrazole-3β-yl chloride has also been carried out against five human tumor cell lines. Doxorubicin is used as a standard drug for the in vitro anti-tumor screening.

  13. The Activity of Menkes Disease Protein ATP7A Is Essential for Redox Balance in Mitochondria.

    PubMed

    Bhattacharjee, Ashima; Yang, Haojun; Duffy, Megan; Robinson, Emily; Conrad-Antoville, Arianrhod; Lu, Ya-Wen; Capps, Tony; Braiterman, Lelita; Wolfgang, Michael; Murphy, Michael P; Yi, Ling; Kaler, Stephen G; Lutsenko, Svetlana; Ralle, Martina

    2016-08-01

    Copper-transporting ATPase ATP7A is essential for mammalian copper homeostasis. Loss of ATP7A activity is associated with fatal Menkes disease and various other pathologies. In cells, ATP7A inactivation disrupts copper transport from the cytosol into the secretory pathway. Using fibroblasts from Menkes disease patients and mouse 3T3-L1 cells with a CRISPR/Cas9-inactivated ATP7A, we demonstrate that ATP7A dysfunction is also damaging to mitochondrial redox balance. In these cells, copper accumulates in nuclei, cytosol, and mitochondria, causing distinct changes in their redox environment. Quantitative imaging of live cells using GRX1-roGFP2 and HyPer sensors reveals highest glutathione oxidation and elevation of H2O2 in mitochondria, whereas the redox environment of nuclei and the cytosol is much less affected. Decreasing the H2O2 levels in mitochondria with MitoQ does not prevent glutathione oxidation; i.e. elevated copper and not H2O2 is a primary cause of glutathione oxidation. Redox misbalance does not significantly affect mitochondrion morphology or the activity of respiratory complex IV but markedly increases cell sensitivity to even mild glutathione depletion, resulting in loss of cell viability. Thus, ATP7A activity protects mitochondria from excessive copper entry, which is deleterious to redox buffers. Mitochondrial redox misbalance could significantly contribute to pathologies associated with ATP7A inactivation in tissues with paradoxical accumulation of copper (i.e. renal epithelia). PMID:27226607

  14. Analysis Of DWPF Sludge Batch 7a (Macrobatch 8) Pour Stream Samples

    SciTech Connect

    Johnson, F. C.; Pareizs, J. M.

    2012-10-24

    The Defense Waste Processing Facility (DWPF) began processing Sludge Batch 7a (SB7a), also referred to as Macrobatch 8 (MB8), in June 2011. SB7a is a blend of the heel of Tank 40 from Sludge Batch 6 (SB6) and the SB7a material that was transferred to Tank 40 from Tank 51. SB7a was processed using Frit 418. During processing of each sludge batch, the DWPF is required to take at least one glass sample to meet the objectives of the Glass Product Control Program (GPCP), which is governed by the DWPF Waste Compliance Plan, and to complete the necessary Production Records so that the final glass product may be disposed of at a Federal Repository. Three pour stream glass samples and two Melter Feed Tank (MFT) slurry samples were collected while processing SB7a. These additional samples were taken during SB7a to understand the impact of antifoam and the melter bubblers on glass redox chemistry. The samples were transferred to the Savannah River National Laboratory (SRNL) where they were analyzed.

  15. ZBTB7A mutations in acute myeloid leukaemia with t(8;21) translocation

    PubMed Central

    Hartmann, Luise; Dutta, Sayantanee; Opatz, Sabrina; Vosberg, Sebastian; Reiter, Katrin; Leubolt, Georg; Metzeler, Klaus H.; Herold, Tobias; Bamopoulos, Stefanos A.; Bräundl, Kathrin; Zellmeier, Evelyn; Ksienzyk, Bianka; Konstandin, Nikola P.; Schneider, Stephanie; Hopfner, Karl-Peter; Graf, Alexander; Krebs, Stefan; Blum, Helmut; Middeke, Jan Moritz; Stölzel, Friedrich; Thiede, Christian; Wolf, Stephan; Bohlander, Stefan K.; Preiss, Caroline; Chen-Wichmann, Linping; Wichmann, Christian; Sauerland, Maria Cristina; Büchner, Thomas; Berdel, Wolfgang E.; Wörmann, Bernhard J.; Braess, Jan; Hiddemann, Wolfgang; Spiekermann, Karsten; Greif, Philipp A.

    2016-01-01

    The t(8;21) translocation is one of the most frequent cytogenetic abnormalities in acute myeloid leukaemia (AML) and results in the RUNX1/RUNX1T1 rearrangement. Despite the causative role of the RUNX1/RUNX1T1 fusion gene in leukaemia initiation, additional genetic lesions are required for disease development. Here we identify recurring ZBTB7A mutations in 23% (13/56) of AML t(8;21) patients, including missense and truncating mutations resulting in alteration or loss of the C-terminal zinc-finger domain of ZBTB7A. The transcription factor ZBTB7A is important for haematopoietic lineage fate decisions and for regulation of glycolysis. On a functional level, we show that ZBTB7A mutations disrupt the transcriptional repressor potential and the anti-proliferative effect of ZBTB7A. The specific association of ZBTB7A mutations with t(8;21) rearranged AML points towards leukaemogenic cooperativity between mutant ZBTB7A and the RUNX1/RUNX1T1 fusion. PMID:27252013

  16. The Activity of Menkes Disease Protein ATP7A Is Essential for Redox Balance in Mitochondria*

    PubMed Central

    Bhattacharjee, Ashima; Yang, Haojun; Duffy, Megan; Robinson, Emily; Conrad-Antoville, Arianrhod; Lu, Ya-Wen; Capps, Tony; Braiterman, Lelita; Wolfgang, Michael; Murphy, Michael P.; Yi, Ling; Kaler, Stephen G.; Lutsenko, Svetlana; Ralle, Martina

    2016-01-01

    Copper-transporting ATPase ATP7A is essential for mammalian copper homeostasis. Loss of ATP7A activity is associated with fatal Menkes disease and various other pathologies. In cells, ATP7A inactivation disrupts copper transport from the cytosol into the secretory pathway. Using fibroblasts from Menkes disease patients and mouse 3T3-L1 cells with a CRISPR/Cas9-inactivated ATP7A, we demonstrate that ATP7A dysfunction is also damaging to mitochondrial redox balance. In these cells, copper accumulates in nuclei, cytosol, and mitochondria, causing distinct changes in their redox environment. Quantitative imaging of live cells using GRX1-roGFP2 and HyPer sensors reveals highest glutathione oxidation and elevation of H2O2 in mitochondria, whereas the redox environment of nuclei and the cytosol is much less affected. Decreasing the H2O2 levels in mitochondria with MitoQ does not prevent glutathione oxidation; i.e. elevated copper and not H2O2 is a primary cause of glutathione oxidation. Redox misbalance does not significantly affect mitochondrion morphology or the activity of respiratory complex IV but markedly increases cell sensitivity to even mild glutathione depletion, resulting in loss of cell viability. Thus, ATP7A activity protects mitochondria from excessive copper entry, which is deleterious to redox buffers. Mitochondrial redox misbalance could significantly contribute to pathologies associated with ATP7A inactivation in tissues with paradoxical accumulation of copper (i.e. renal epithelia). PMID:27226607

  17. Diverse roles for Wnt7a in ventral midbrain neurogenesis and dopaminergic axon morphogenesis.

    PubMed

    Fernando, Chathurini V; Kele, Julianna; Bye, Christopher R; Niclis, Jonathan C; Alsanie, Walaa; Blakely, Brette D; Stenman, Jan; Turner, Brad J; Parish, Clare L

    2014-09-01

    During development of the central nervous system, trophic, together with genetic, cues dictate the balance between cellular proliferation and differentiation. Subsequent to the birth of new neurons, additional intrinsic and extrinsic signals regulate the connectivity of these cells. While a number of regulators of ventral midbrain (VM) neurogenesis and dopaminergic (DA) axon guidance are known, we identify a number of novel roles for the secreted glycoprotein, Wnt7a, in this context. We demonstrate a temporal and spatial expression of Wnt7a in the VM, indicative of roles in neurogenesis, differentiation, and axonal growth and guidance. In primary VM cultures, and validated in Wnt7a-deficient mice, we show that the early expression within the VM is important for regulating VM progenitor proliferation, cell cycle progression, and cell survival, thereby dictating the number of midbrain Nurr1 precursors and DA neurons. During early development of the midbrain DA pathways, Wnt7a promotes axonal elongation and repels DA neurites out of the midbrain. Later, Wnt7a expression in the VM midline suggests a role in preventing axonal crossing while expression in regions flanking the medial forebrain bundle (thalamus and hypothalamus) ensured appropriate trajectory of DA axons en route to their forebrain targets. We show that the effects of Wnt7a in VM development are mediated, at least in part, by the β-catenin/canonical pathways. Together, these findings identify Wnt7a as a new regulator of VM neurogenesis and DA axon growth and guidance.

  18. Unexpected role of the copper transporter ATP7A in PDGF-induced vascular smooth

    SciTech Connect

    Ashino, T.; Varadarajan, S.; Urao, N.; Oshikawa, J.; Chen, G. -F.; Wang, H.; Huo, Y.; Finney, L.; Vogt, S.; McKinney, R. D.; Maryon, E. B.; Kaplan, J. H.; Ushio-Fukai, M.; Fukai, T.

    2010-09-09

    Copper, an essential nutrient, has been implicated in vascular remodeling and atherosclerosis with unknown mechanism. Bioavailability of intracellular copper is regulated not only by the copper importer CTR1 (copper transporter 1) but also by the copper exporter ATP7A (Menkes ATPase), whose function is achieved through copper-dependent translocation from trans-Golgi network (TGN). Platelet-derived growth factor (PDGF) promotes vascular smooth muscle cell (VSMC) migration, a key component of neointimal formation. To determine the role of copper transporter ATP7A in PDGF-induced VSMC migration. Depletion of ATP7A inhibited VSMC migration in response to PDGF or wound scratch in a CTR1/copper-dependent manner. PDGF stimulation promoted ATP7A translocation from the TGN to lipid rafts, which localized at the leading edge, where it colocalized with PDGF receptor and Rac1, in migrating VSMCs. Mechanistically, ATP7A small interfering RNA or CTR small interfering RNA prevented PDGF-induced Rac1 translocation to the leading edge, thereby inhibiting lamellipodia formation. In addition, ATP7A depletion prevented a PDGF-induced decrease in copper level and secretory copper enzyme precursor prolysyl oxidase (Pro-LOX) in lipid raft fraction, as well as PDGF-induced increase in LOX activity. In vivo, ATP7A expression was markedly increased and copper accumulation was observed by synchrotron-based x-ray fluorescence microscopy at neointimal VSMCs in wire injury model. These findings suggest that ATP7A plays an important role in copper-dependent PDGF-stimulated VSMC migration via recruiting Rac1 to lipid rafts at the leading edge, as well as regulating LOX activity. This may contribute to neointimal formation after vascular injury. Our findings provide insight into ATP7A as a novel therapeutic target for vascular remodeling and atherosclerosis.

  19. THE SLUDGE BATCH 7A GLASS VARIABILITY STUDY WITH FRIT 418 AND FRIT 702

    SciTech Connect

    Peeler, D; Edwards, T

    2011-03-24

    The Defense Waste Processing Facility (DWPF) is preparing to initiate processing of Sludge Batch 7a (SB7a) in May 2011. To support qualification of SB7a, the Savannah River National Laboratory (SRNL) was requested to execute a variability study (VS) to assess the applicability of the current Product Composition Control System (PCCS) durability models for the Frit 418-SB7a compositional region of interest. The objective of this study was to demonstrate applicability of the current durability models to the SB7a compositional region of interest and acceptability of the SB7a glasses with respect to the Environmental Assessment (EA) glass in terms of durability as defined by the Product Consistency Test (PCT). To support programmatic objectives, twenty-eight SB7a glasses were selected based on the nominal sludge projections used to support the frit recommendation. Twenty-three of the SB7a VS glasses were based on the use of Frit 418, while 5 glasses were based on the use of Frit 702. Frit 702 was also identified as a viable candidate for SB7a, especially if SO{sub 4} concentrations are found to be higher than anticipated. Frit 702 has shown a higher SO{sub 4} retention capability as compared to Frit 418. With respect to acceptability, the PCT results of the SB7a-VS glasses are acceptable relative to the EA glass regardless of thermal history (quenched or canister centerline cooled) or compositional view (target or measured). More specifically, all of the SB7a glasses have normalized boron release values (NL [B]) less than 0.9 g/L as compared to the benchmark NL [B] value for EA of 16.695 g/L. With respect to the applicability of the current durability models to the SB7a VS compositional region of interest, all of the study glasses (based on target compositions) lie within the 95% confidence intervals of the model predictions. When model applicability is based on the measured compositions, all of the SB7a VS glasses are predictable with the exception of SB7aVS-02 and SB7

  20. miR-Let7A Modulates Autophagy Induction in LPS-Activated Microglia

    PubMed Central

    Song, Juhyun; Oh, Yumi

    2015-01-01

    Microglia regulate the secretion of various immunomediators in central nervous system diseases. Microglial autophagy is the crucial process for cell's survival and cytokine productions. Recent studies have reported that several microRNAs are involved in the autophagy system. miR-Let7A is such a microRNA that plays a role in various inflammation responses, and is magnified as a key modulator particularly in the autophagy system. In present study, we investigated whether miR-Let7A is involved in autophagy in activating microglia. Overexpression of miR-Let7A in LPS-stimulated BV2 microglial cells promoted the induction of the autophagy related factors such as LC3II, Beclin1, and ATG3. Our results suggest a potential role of miR-Let7A in the autophagy process of microglia during CNS inflammation. PMID:26113790

  1. Let7a inhibits the growth of endometrial carcinoma cells by targeting Aurora-B.

    PubMed

    Liu, Ping; Qi, Meiyan; Ma, Chengbin; Lao, Guoying; Liu, Yu; Liu, Yan; Liu, Yingzi

    2013-08-19

    MicroRNAs negatively regulate target gene expression at the post-transcriptional level during carcinogenesis. Recent advances revealed that the expression levels of several miRNAs are up- or down-regulated in endometrial carcinoma (EC). Here we identify dysregulated miRNAs in EC and we elucidate the essential role of let-7a. The expression of 86 miRNAs in EC was found to be different from adjacent normal endometrial tissues. Moreover, miR-let-7 members are down-regulated in EC and let-7 miRNAs are highly associated with endometrial cancer. A functional investigation revealed that let-7a suppressed proliferation of HeLa cells by targeting Aurora-B. Let-7a also antagonizes Aurora-B functions in promoting carcinoma cell proliferation by down-regulating Aurora-B protein level. Let-7a could be applied for gene therapy against endometrial carcinogenesis. PMID:23769985

  2. MicroRNA-Let-7a regulates the function of microglia in inflammation.

    PubMed

    Cho, Kyoung Joo; Song, Juhyun; Oh, Yumi; Lee, Jong Eun

    2015-09-01

    Microglia have multiple functions in cerebrovascular and neurodegenerative diseases. Regulation of microglial function during inflammatory stress is important for treatment of central nervous system (CNS) diseases because microglia secrete various substances that affect neurons and glia. MicroRNA-Let-7a (miR-Let-7a) is a tumor suppressor miRNA that has been reported to target transcripts that encode proteins involved in apoptosis. In the present study, we examined the essential role of miR-Let-7a in inflammatory stress by over-expressing miR-Let-7a to investigate its role in determining the BV2 microglial phenotype, a cell line often used as a model of activated microglia. We found that inflammatory factors and Reactive Oxygen Species (ROS) production levels were altered according to miR-Let-7a expression level as measured by Western blot analysis, reverse transcription PCR, quantitative real time PCR, the measurement of nitrite (indicative of the nitric oxide (NO) pathway), and immunocytochemistry (ICC). Our results suggest that miR-Let-7a is involved in the function of microglia in the setting of inflammatory injury. In response to inflammation, miR-Let-7a participates in the reduction of nitrite production and the expression of inducible nitric oxide synthase (iNOS), interleukin (IL)-6 and is involved in increased expression of brain derived neurotrophic factor (BDNF), interleukin (IL)-10, and IL-4 in microglia. Thus, miRNA-Let-7a could act as a regulator of the function of microglia in inflammation.

  3. PDZD7-MYO7A complex identified in enriched stereocilia membranes

    PubMed Central

    Morgan, Clive P; Krey, Jocelyn F; Grati, M'hamed; Zhao, Bo; Fallen, Shannon; Kannan-Sundhari, Abhiraami; Liu, Xue Zhong; Choi, Dongseok; Müller, Ulrich; Barr-Gillespie, Peter G

    2016-01-01

    While more than 70 genes have been linked to deafness, most of which are expressed in mechanosensory hair cells of the inner ear, a challenge has been to link these genes into molecular pathways. One example is Myo7a (myosin VIIA), in which deafness mutations affect the development and function of the mechanically sensitive stereocilia of hair cells. We describe here a procedure for the isolation of low-abundance protein complexes from stereocilia membrane fractions. Using this procedure, combined with identification and quantitation of proteins with mass spectrometry, we demonstrate that MYO7A forms a complex with PDZD7, a paralog of USH1C and DFNB31. MYO7A and PDZD7 interact in tissue-culture cells, and co-localize to the ankle-link region of stereocilia in wild-type but not Myo7a mutant mice. Our data thus describe a new paradigm for the interrogation of low-abundance protein complexes in hair cell stereocilia and establish an unanticipated link between MYO7A and PDZD7. DOI: http://dx.doi.org/10.7554/eLife.18312.001 PMID:27525485

  4. Let-7a inhibits migration of melanoma cells via down-regulation of HMGA2 expression

    PubMed Central

    Hou, Xiaocan; Wan, Wencui; Wang, Jing; Li, Mingzhe; Wang, Yiwen; Yao, Yaobing; Feng, Lihong; Jing, Lijun; Lu, Hong; Jia, Yanjie; Peng, Tao

    2016-01-01

    This study aimed to investigate the effects of exosomes derived from BM-MSCs transduced with let-7a on B16f10 cells and BM-MSCs. BM-MSCs were transduced with let-7a and the exosomes of them were isolated for further culture of B16f10 cells and BM-MSCs. The migration of B16f10 cells were detected by transwell, proliferation of B16f10 cells and BM-MSCs was examined by MTT method, HMGA2 expression was measured by western blot. In addition, the let-7a secreted level in exosomes and IGF level were measured by RT-PCR and ELISA respectively. Our results showed that the level of let-7a in exosomes derived from Let-7a-transducted BM-MSCs was increased after treated by exosomes. HMGA2 in B16f10 cells was down-regulated and cell survival rate of BM-MSCs was decreased. However, neither cell survival rate of B16f10 cells nor IGF-1 secreted by B16f10 cells in different groups had significant differences. In conclusion, Let-7a contained in exosomes can inhibit the migration of Melanoma cells and inhibit the proliferation of BM-MSCs. PMID:27725848

  5. Naringenin Ameliorated Kidney Injury through Let-7a/TGFBR1 Signaling in Diabetic Nephropathy

    PubMed Central

    Yan, Ning; Peng, Rui; Li, Hongmei; Liu, Handeng; Peng, Huimin; Sun, Yan; Wu, Tianhui; Chen, Lei; Duan, Qingrui; Sun, Yixuan; Zhou, Qin; Wei, Lijiang

    2016-01-01

    Diabetic nephropathy (DN) is one of the most common complications of diabetes mellitus (DM). However, the exact mechanism is not clearly understood. In this study, our results showed that 24 h urinary protein, kidney index, and glomerular area were decreased, while creatinine clearance ratio was increased in DN rats when the rats were treated with NAR 50 mg/d for 6 weeks. Mesangial cell (MMCs) proliferation was inhibited in the NAR group by 3,(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), and the cell cycle analysis showed that cells stayed in G2 phase in NAR group. And NAR treatment attenuated the deposition of ECM in DN rats and MMCs. Moreover, our data showed that let-7a was downexpressed in both DN rats and MMCs under high glucose condition. Surprisingly, NAR affected the expressions of Col4 and FN through upregulating let-7a in MMCs. In addition, we found that let-7a negatively regulated the expression of transforming growth factor-β1 receptor 1 (TGFBR1), and TGFBR1 was required for the let-7a-mediated downregulation of TGF-β1/smad signaling. Interestingly, NAR inhibited TGF-β1/smads signaling activation by upregulating let-7a. Therefore, our findings indicated that NAR ameliorated kidney injury by regulating let-7a/TGFBR1 signaling. PMID:27446963

  6. PDZD7-MYO7A complex identified in enriched stereocilia membranes.

    PubMed

    Morgan, Clive P; Krey, Jocelyn F; Grati, M'hamed; Zhao, Bo; Fallen, Shannon; Kannan-Sundhari, Abhiraami; Liu, Xue Zhong; Choi, Dongseok; Müller, Ulrich; Barr-Gillespie, Peter G

    2016-01-01

    While more than 70 genes have been linked to deafness, most of which are expressed in mechanosensory hair cells of the inner ear, a challenge has been to link these genes into molecular pathways. One example is Myo7a (myosin VIIA), in which deafness mutations affect the development and function of the mechanically sensitive stereocilia of hair cells. We describe here a procedure for the isolation of low-abundance protein complexes from stereocilia membrane fractions. Using this procedure, combined with identification and quantitation of proteins with mass spectrometry, we demonstrate that MYO7A forms a complex with PDZD7, a paralog of USH1C and DFNB31. MYO7A and PDZD7 interact in tissue-culture cells, and co-localize to the ankle-link region of stereocilia in wild-type but not Myo7a mutant mice. Our data thus describe a new paradigm for the interrogation of low-abundance protein complexes in hair cell stereocilia and establish an unanticipated link between MYO7A and PDZD7. PMID:27525485

  7. New metal-organic frameworks of [M(C{sub 6}H{sub 5}O{sub 7})(C{sub 6}H{sub 6}O{sub 7})(C{sub 6}H{sub 7}O{sub 7})(H{sub 2}O)] . H{sub 2}O (M=La, Ce) and [Ce{sub 2}(C{sub 2}O{sub 4})(C{sub 6}H{sub 6}O{sub 7}){sub 2}] . 4H{sub 2}O

    SciTech Connect

    Weng Shengfeng; Wang, Yun-Hsin; Lee, Chi-Shen

    2012-04-15

    Two novel materials, [M(C{sub 6}H{sub 5}O{sub 7})(C{sub 6}H{sub 6}O{sub 7})(C{sub 6}H{sub 7}O{sub 7})(H{sub 2}O)] . H{sub 2}O (M=La(1a), Ce(1b)) and [Ce{sub 2}(C{sub 2}O{sub 4})(C{sub 6}H{sub 6}O{sub 7}){sub 2}] . 4H{sub 2}O (2), with a metal-organic framework (MOF) were prepared with hydrothermal reactions and characterized with photoluminescence, magnetic susceptibility, thermogravimetric analysis and X-ray powder diffraction in situ. The crystal structures were determined by single-crystal X-ray diffraction. Compound 1 crystallized in triclinic space group P1-bar (No. 2); compound 2 crystallized in monoclinic space group P2{sub 1}/c (No. 14). The structure of 1 is built from a 1D MOF, composed of deprotonated citric ligands of three kinds. Compound 2 contains a 2D MOF structure consisting of citrate and oxalate ligands; the oxalate ligand arose from the decomposition in situ of citric acid in the presence of Cu{sup II} ions. Photoluminescence spectra of compounds 1b and 2 revealed transitions between the 5d{sup 1} excited state and two levels of the 4f{sup 1} ground state ({sup 2}F{sub 5/2} and {sup 2}F{sub 7/2}). Compounds 1b and 2 containing Ce{sup III} ion exhibit a paramagnetic property with weak antiferromagnetic interactions between the two adjacent magnetic centers. - Graphical Abstract: [M(C{sub 6}H{sub 5}O{sub 7})(C{sub 6}H{sub 6}O{sub 7})(C{sub 6}H{sub 7}O{sub 7})(H{sub 2}O)] . H{sub 2}O (M=La(1a), Ce(1b)) and [Ce{sub 2}(C{sub 2}O{sub 4})(C{sub 6}H{sub 6}O{sub 7}){sub 2}] . 4H{sub 2}O (2)-with 1D and 2D structures were synthesized and characterized. Highlights: Black-Right-Pointing-Pointer Two MOF - [M(C{sub 6}H{sub 5}O{sub 7})(C{sub 6}H{sub 6}O{sub 7})(C{sub 6}H{sub 7}O{sub 7})(H{sub 2}O)] . H{sub 2}O (M=La(1a), Ce(1b)) and [Ce{sub 2}(C{sub 2}O{sub 4})(C{sub 6}H{sub 6}O{sub 7}){sub 2}] . 4H{sub 2}O (2) - with 1D and 2D structures. Black-Right-Pointing-Pointer The adjacent chains of the 1D framework were correlated with each other through an oxalate

  8. Derivatives of 1,5-diamino-1H-tetrazole: a new family of energetic heterocyclic-based salts.

    PubMed

    Gálvez-Ruiz, Juan Carlos; Holl, Gerhard; Karaghiosoff, Konstantin; Klapötke, Thomas M; Löhnwitz, Karolin; Mayer, Peter; Nöth, Heinrich; Polborn, Kurt; Rohbogner, Christoph J; Suter, Max; Weigand, Jan J

    2005-06-13

    1,5-Diamino-1H-tetrazole (2, DAT) can easily be protonated by reaction with strong mineral acids, yielding the poorly investigated 1,5-diaminotetrazolium nitrate (2a) and perchlorate (2b). A new synthesis for 2 is introduced that avoids lead azide as a hazardous byproduct. The reaction of 1,5-diamino-1H-tetrazole with iodomethane (7a) followed by the metathesis of the iodide (7a) with silver nitrate (7b), silver dinitramide (7c), or silver azide (7d) leads to a new family of heterocyclic-based salts. In all cases, stable salts were obtained and fully characterized by vibrational (IR, Raman) spectroscopy, multinuclear NMR spectroscopy, mass spectrometry, elemental analysis, X-ray structure determination, and initial safety testing (impact and friction sensitivity). Most of the salts exhibit good thermal stabilities, and both the perchlorate (2b) and the dinitramide (7c) have melting points well below 100 degrees C, yet high decomposition onsets, defining them as new (7c), highly energetic ionic liquids. Preliminary sensitivity testing of the crystalline compounds indicates rather low impact sensitivities for all compounds, the highest being that of the perchlorate (2b) and the dinitramide (7c) with a value of 7 J. In contrast, the friction sensitivities of the perchlorate (2b, 60 N) and the dinitramide (7c, 24 N) are relatively high. The enthalpies of combustion (Delta(c)H degrees ) of 7b-d were determined experimentally using oxygen bomb calorimetry: Delta(c)H degrees (7b) = -2456 cal g(-)(1), Delta(c)H degrees (7c) = -2135 cal g(-)(1), and Delta(c)H degrees (7d) = -3594 cal g(-)(1). The standard enthalpies of formation (Delta(f)H degrees ) of 7b-d were obtained on the basis of quantum chemical computations using the G2 (G3) method: Delta(f)H degrees (7b) = 41.7 (41.2) kcal mol(-)(1), Delta(f)H degrees (7c) = 92.1 (91.1) kcal mol(-)(1), and Delta(f)H degrees (7d) = 161.6 (161.5) kcal mol(-)(1). The detonation velocities (D) and detonation pressures (P) of 2b and 7b

  9. Derivatives of 1,5-diamino-1H-tetrazole: a new family of energetic heterocyclic-based salts.

    PubMed

    Gálvez-Ruiz, Juan Carlos; Holl, Gerhard; Karaghiosoff, Konstantin; Klapötke, Thomas M; Löhnwitz, Karolin; Mayer, Peter; Nöth, Heinrich; Polborn, Kurt; Rohbogner, Christoph J; Suter, Max; Weigand, Jan J

    2005-06-13

    1,5-Diamino-1H-tetrazole (2, DAT) can easily be protonated by reaction with strong mineral acids, yielding the poorly investigated 1,5-diaminotetrazolium nitrate (2a) and perchlorate (2b). A new synthesis for 2 is introduced that avoids lead azide as a hazardous byproduct. The reaction of 1,5-diamino-1H-tetrazole with iodomethane (7a) followed by the metathesis of the iodide (7a) with silver nitrate (7b), silver dinitramide (7c), or silver azide (7d) leads to a new family of heterocyclic-based salts. In all cases, stable salts were obtained and fully characterized by vibrational (IR, Raman) spectroscopy, multinuclear NMR spectroscopy, mass spectrometry, elemental analysis, X-ray structure determination, and initial safety testing (impact and friction sensitivity). Most of the salts exhibit good thermal stabilities, and both the perchlorate (2b) and the dinitramide (7c) have melting points well below 100 degrees C, yet high decomposition onsets, defining them as new (7c), highly energetic ionic liquids. Preliminary sensitivity testing of the crystalline compounds indicates rather low impact sensitivities for all compounds, the highest being that of the perchlorate (2b) and the dinitramide (7c) with a value of 7 J. In contrast, the friction sensitivities of the perchlorate (2b, 60 N) and the dinitramide (7c, 24 N) are relatively high. The enthalpies of combustion (Delta(c)H degrees ) of 7b-d were determined experimentally using oxygen bomb calorimetry: Delta(c)H degrees (7b) = -2456 cal g(-)(1), Delta(c)H degrees (7c) = -2135 cal g(-)(1), and Delta(c)H degrees (7d) = -3594 cal g(-)(1). The standard enthalpies of formation (Delta(f)H degrees ) of 7b-d were obtained on the basis of quantum chemical computations using the G2 (G3) method: Delta(f)H degrees (7b) = 41.7 (41.2) kcal mol(-)(1), Delta(f)H degrees (7c) = 92.1 (91.1) kcal mol(-)(1), and Delta(f)H degrees (7d) = 161.6 (161.5) kcal mol(-)(1). The detonation velocities (D) and detonation pressures (P) of 2b and 7b

  10. WNT7A Regulation by miR-15b in Ovarian Cancer.

    PubMed

    MacLean, James A; King, Mandy L; Okuda, Hiroshi; Hayashi, Kanako

    2016-01-01

    WNT signaling is well known to play an important role in the regulation of development, cell proliferation and cell differentiation in a wide variety of normal and cancerous tissues. Despite the wealth of knowledge concerning when and where various WNT genes are expressed and downstream events under their control, there is surprisingly little published evidence of how they are regulated. We have recently reported that aberrant WNT7A is observed in serous ovarian carcinomas, and WNT7A is the sole ligand accelerating ovarian tumor progression through CTNNB1 (β-catenin)/TCF signaling in the absence of CTNNB1 mutations. In the present study, we report that WNT7A is a direct target of miR-15b in ovarian cancer. We showed that a luciferase reporter containing the putative binding site of miR-15b in the WNT7A 3'-UTR was significantly repressed by miR-15b. Mutation of the putative binding site of miR-15b in the WNT7A 3'-UTR restored luciferase activity. Furthermore, miR-15b was able to repress increased levels of TOPFLASH activity by WNT7A, but not those induced by S33Y. Additionally, miR-15b dose-dependently decreased WNT7A expression. When we evaluated the prognostic impact of WNT7A and miR-15b expression using TCGA datasets, a significant inverse correlation in which high-expression of WNT7A and low-expression of miR-15b was associated with reduced survival rates of ovarian cancer patients. Treatment with decitabine dose-dependently increased miR-15b expression, and silencing of DNMT1 significantly increased miR-15b expression. These results suggest that WNT7A is post-transcriptionally regulated by miR-15b, which could be down-regulated by promoter hypermethylation, potentially via DNMT1, in ovarian cancer. PMID:27195958

  11. Structural Basis of Substrate Recognition by Aldehyde Dehydrogenase 7A1

    PubMed Central

    2016-01-01

    Aldehyde dehydrogenase 7A1 (ALDH7A1) is part of lysine catabolism and catalyzes the NAD+-dependent oxidation of α-aminoadipate semialdehyde to α-aminoadipate. Herein, we describe a structural study of human ALDH7A1 focused on substrate recognition. Five crystal structures and small-angle X-ray scattering data are reported, including the first crystal structure of any ALDH7 family member complexed with α-aminoadipate. The product binds with the ε-carboxylate in the oxyanion hole, the aliphatic chain packed into an aromatic box, and the distal end of the product anchored by electrostatic interactions with five conserved residues. This binding mode resembles that of glutamate bound to the proline catabolic enzyme ALDH4A1. Analysis of ALDH7A1 and ALDH4A1 structures suggests key interactions that underlie substrate discrimination. Structures of apo ALDH7A1 reveal dramatic conformational differences from the product complex. Product binding is associated with a 16 Å movement of the C-terminus into the active site, which stabilizes the active conformation of the aldehyde substrate anchor loop. The fact that the C-terminus is part of the active site was hitherto unknown. Interestingly, the C-terminus and aldehyde anchor loop are disordered in a new tetragonal crystal form of the apoenzyme, implying that these parts of the enzyme are highly flexible. Our results suggest that the active site of ALDH7A1 is disassembled when the aldehyde site is vacant, and the C-terminus is a mobile element that forms quaternary structural interactions that aid aldehyde binding. These results are relevant to the c.1512delG genetic deletion associated with pyridoxine-dependent epilepsy, which alters the C-terminus of ALDH7A1. PMID:26260980

  12. Exon duplications in the ATP7A gene: Frequency and Transcriptional Behaviour

    PubMed Central

    2011-01-01

    Background Menkes disease (MD) is an X-linked, fatal neurodegenerative disorder of copper metabolism, caused by mutations in the ATP7A gene. Thirty-three Menkes patients in whom no mutation had been detected with standard diagnostic tools were screened for exon duplications in the ATP7A gene. Methods The ATP7A gene was screened for exon duplications using multiplex ligation-dependent probe amplification (MLPA). The expression level of ATP7A was investigated by real-time PCR and detailed analysis of the ATP7A mRNA was performed by RT-PCR followed by sequencing. In order to investigate whether the identified duplicated fragments originated from a single or from two different X-chromosomes, polymorphic markers located in the duplicated fragments were analyzed. Results Partial ATP7A gene duplication was identified in 20 unrelated patients including one patient with Occipital Horn Syndrome (OHS). Duplications in the ATP7A gene are estimated from our material to be the disease causing mutation in 4% of the Menkes disease patients. The duplicated regions consist of between 2 and 15 exons. In at least one of the cases, the duplication was due to an intra-chromosomal event. Characterization of the ATP7A mRNA transcripts in 11 patients revealed that the duplications were organized in tandem, in a head to tail direction. The reading frame was disrupted in all 11 cases. Small amounts of wild-type transcript were found in all patients as a result of exon-skipping events occurring in the duplicated regions. In the OHS patient with a duplication of exon 3 and 4, the duplicated out-of-frame transcript coexists with an almost equally represented wild-type transcript, presumably leading to the milder phenotype. Conclusions In general, patients with duplication of only 2 exons exhibit a milder phenotype as compared to patients with duplication of more than 2 exons. This study provides insight into exon duplications in the ATP7A gene. PMID:22074552

  13. Let7a involves in neural stem cell differentiation relating with TLX level.

    PubMed

    Song, Juhyun; Cho, Kyoung Joo; Oh, Yumi; Lee, Jong Eun

    2015-07-10

    Neural stem cells (NSCs) have the potential for differentiation into neurons known as a groundbreaking therapeutic solution for central nervous system (CNS) diseases. To resolve the therapeutic efficiency of NSCs, recent researchers have focused on the study on microRNA's role in CNS. Some micro RNAs have been reported significant functions in NSC self-renewal and differentiation through the post-transcriptional regulation of neurogenesis genes. MicroRNA-Let7a (Let7a) has known as the regulator of diverse cellular mechanisms including cell differentiation and proliferation. In present study, we investigated whether Let7a regulates NSC differentiation by targeting the nuclear receptor TLX, which is an essential regulator of NSC self-renewal, proliferation and differentiation. We performed the following experiments: western blot analysis, TaqMan assay, RT-PCR, and immunocytochemistry to confirm the alteration of NSCs. Our data showed that let7a play important roles in controlling NSC fate determination. Thus, manipulating Let-7A and TLX could be a novel strategy to enhance the efficiency of NSC's neuronal differentiation for CNS disorders.

  14. Genome sequence of the Lotus spp. microsymbiont Mesorhizobium loti strain R7A

    PubMed Central

    2014-01-01

    Mesorhizobium loti strain R7A was isolated in 1993 in Lammermoor, Otago, New Zealand from a Lotus corniculatus root nodule and is a reisolate of the inoculant strain ICMP3153 (NZP2238) used at the site. R7A is an aerobic, Gram-negative, non-spore-forming rod. The symbiotic genes in the strain are carried on a 502-kb integrative and conjugative element known as the symbiosis island or ICEMlSymR7A. M. loti is the microsymbiont of the model legume Lotus japonicus and strain R7A has been used extensively in studies of the plant-microbe interaction. This report reveals that the genome of M. loti strain R7A does not harbor any plasmids and contains a single scaffold of size 6,529,530 bp which encodes 6,323 protein-coding genes and 75 RNA-only encoding genes. This rhizobial genome is one of 100 sequenced as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project. PMID:25780499

  15. Suppression of MicroRNA let-7a Expression by Agmatine Regulates Neural Stem Cell Differentiation

    PubMed Central

    Song, Juhyun; Oh, Yumi; Kim, Jong Youl; Cho, Kyoung Joo

    2016-01-01

    Purpose Neural stem cells (NSCs) effectively reverse some severe central nervous system (CNS) disorders, due to their ability to differentiate into neurons. Agmatine, a biogenic amine, has cellular protective effects and contributes to cellular proliferation and differentiation in the CNS. Recent studies have elucidated the function of microRNA let-7a (let-7a) as a regulator of cell differentiation with roles in regulating genes associated with CNS neurogenesis. Materials and Methods This study aimed to investigate whether agmatine modulates the expression of crucial regulators of NSC differentiation including DCX, TLX, c-Myc, and ERK by controlling let-7a expression. Results Our data suggest that high levels of let-7a promoted the expression of TLX and c-Myc, as well as repressed DCX and ERK expression. In addition, agmatine attenuated expression of TLX and increased expression of ERK by negatively regulating let-7a. Conclusion Our study therefore enhances the present understanding of the therapeutic potential of NSCs in CNS disorders. PMID:27593875

  16. Wnt7a stimulates myogenic stem cell motility and engraftment resulting in improved muscle strength.

    PubMed

    Bentzinger, C Florian; von Maltzahn, Julia; Dumont, Nicolas A; Stark, Danny A; Wang, Yu Xin; Nhan, Kevin; Frenette, Jérôme; Cornelison, D D W; Rudnicki, Michael A

    2014-04-14

    Wnt7a/Fzd7 signaling stimulates skeletal muscle growth and repair by inducing the symmetric expansion of satellite stem cells through the planar cell polarity pathway and by activating the Akt/mTOR growth pathway in muscle fibers. Here we describe a third level of activity where Wnt7a/Fzd7 increases the polarity and directional migration of mouse satellite cells and human myogenic progenitors through activation of Dvl2 and the small GTPase Rac1. Importantly, these effects can be exploited to potentiate the outcome of myogenic cell transplantation into dystrophic muscles. We observed that a short Wnt7a treatment markedly stimulated tissue dispersal and engraftment, leading to significantly improved muscle function. Moreover, myofibers at distal sites that fused with Wnt7a-treated cells were hypertrophic, suggesting that the transplanted cells deliver activated Wnt7a/Fzd7 signaling complexes to recipient myofibers. Taken together, we describe a viable and effective ex vivo cell modulation process that profoundly enhances the efficacy of stem cell therapy for skeletal muscle.

  17. Test and evaluation document for DOT Specification 7A type A packaging. Volume 1

    SciTech Connect

    Kelly, D L

    1997-08-04

    The US Department of Energy (DOE) has been conducting, through several of its operating contractors, an evaluation and testing program to qualify Type A radioactive material packagings per US Department of Transportation (DOT) Specification 7A (DOT-7A) of the Code of Federal Regulations (CFR), Title 49, Part 178 (49 CFR 178). This document summarizes the evaluation and testing performed for all of the packagings successfully qualified in this program. This document supersedes DOE Evaluation Document for DOT-7A Type A Packaging (Edling 1987), originally issued in 1987 by Monsanto Research Corporation Mound Laboratory (MLM), Miamisburg, Ohio, for the Department of Energy, Security Evaluation Program (I)P-4. Mound Laboratory issued four revisions to the document between November 1988 and December 1989. In September 1989, the program was transferred to Westinghouse Hanford Company (Westinghouse Hanford) in Richland, Washington. One additional revision was issued in March 1990 by Westinghouse Hanford. This revision reflects the earlier material and incorporates a number of changes. Evaluation and testing activities on 1208 three DOT-7A Program Dockets resulted in the qualification of three new packaging configurations, which are incorporated herein and summarized. This document presents approximately 300 different packagings that have been determined to meet the requirements for a DOT-7A, type A packaging per 49 CFR 178.350.

  18. The respiratory chain supercomplex organization is independent of COX7a2l isoforms.

    PubMed

    Mourier, Arnaud; Matic, Stanka; Ruzzenente, Benedetta; Larsson, Nils-Göran; Milenkovic, Dusanka

    2014-12-01

    The organization of individual respiratory chain complexes into supercomplexes or respirasomes has attracted great interest because of the implications for cellular energy conversion. Recently, it was reported that commonly used mouse strains harbor a short COX7a2l (SCAFI) gene isoform that supposedly precludes the formation of complex IV-containing supercomplexes. This claim potentially has serious implications for numerous mouse studies addressing important topics in metabolism, including adaptation to space flights. Using several complementary experimental approaches, we show that mice with the short COX7a2l isoform have normal biogenesis and steady-state levels of complex IV-containing supercomplexes and consequently have normal respiratory chain function. Furthermore, we use a mouse knockout of Lrpprc and show that loss of complex IV compromises respirasome formation. We conclude that the presence of the short COX7a2l isoform in the commonly used C57BL/6 mouse strains does not prevent their use in metabolism research.

  19. Translational research investigations on ATP7A, an important human copper ATPase

    PubMed Central

    Kaler, Stephen G.

    2014-01-01

    In the more than 40 years since copper deficiency was delineated in pediatric subjects with Menkes disease, remarkable advances in our understanding of the clinical, biochemical, and molecular aspects of the human copper transporter ATP7A have emerged. Mutations in the gene encoding this multitasking molecule are now implicated in at least two other distinctive phenotypes: occipital horn syndrome and ATP7A-related isolated distal motor neuropathy. Several other novel inherited disorders of copper metabolism have been identified in the past several years, aided by advances in human gene mapping and sequencing. In this paper, I review the history and evolution of our understanding of disorders caused by impaired ATP7A function, and outline future challenges. PMID:24735419

  20. Refolding and purification of recombinant human PDE7A expressed in Escherichia coli as inclusion bodies.

    PubMed

    Richter, Wito; Hermsdorf, Thomas; Kronbach, Thomas; Dettmer, Dietrich

    2002-06-01

    We have investigated the refolding and purification of the catalytic domain of human 3',5'-cyclic nucleotide phosphodiesterase 7A1 (PDE7A1) expressed in Escherichia coli. A cDNA encoding an N-terminal-truncated PDE7A1(147-482-His) was amplified by RT-PCR from human peripheral blood cells and inserted into the vector pET21-C for bacterial expression of the enzyme fused to a C-terminal His-tag. The PDE was found to be expressed in the form of inclusion bodies which could be refolded to an active enzyme in buffer containing high concentrations of arginine hydrochloride, ethylene glycol, and magnesium chloride at pH 8.5. The PDE7A1(147-482-His) construct could be purified after dialysis and concentration steps by either Zn2+-IDA-Sepharose chromatography or ResourceQ ion-exchange chromatography to homogeneity. In comparison to the metal-chelate column, the ResourceQ purification resulted in a distinctly better yield and enrichment of the protein. Both the Vmax (0.46 micromol. min(-1). mg(-1) ) and the K(m) (0.1 microM) of the purified enzyme were found to be comparable with published data for native or recombinant catalytically active expressed PDE7A1. Using SDS/PAGE, a molecular mass of 39 kDa was determined (theoretical value 38.783 kDa). As known from several other mammalian PDEs, size-exclusion chromatography using refolded PDE7A1(147-482-His) indicated the formation of dimers. The purified enzyme was soluble at concentrations up to 100 microg/ml. A further increase of protein concentration resulted, however, in precipitation of the enzyme.

  1. Xylan oligosaccharides and cellobiohydrolase I (TrCel7A) interaction and effect on activity

    PubMed Central

    2011-01-01

    Background The well-studied cellulase mixture secreted by Trichoderma reesei (anamorph to Hypocrea jecorina) contains two cellobiohydolases (CBHs), cellobiohydrolase I (TrCel7A) and cellobiohydrolase II (TrCeI6A), that are core enzymes for the solubilisation of cellulose. This has attracted significant research interest because of the role of the CBHs in the conversion of biomass to fermentable sugars. However, the CHBs are notoriously slow and susceptible to inhibition, which presents a challenge for the commercial utilisation of biomass. The xylans and xylan fragments that are also present in the biomass have been suggested repeatedly as one cause of the reduced activity of CHBs. Yet, the extent and mechanisms of this inhibition remain poorly elucidated. Therefore, we studied xylan oligosaccharides (XOSs) of variable lengths with respect to their binding and inhibition of both TrCel7A and an enzyme variant without the cellulose-binding domain (CBM). Results We studied the binding of XOSs to TrCel7A by isothermal titration calorimetry. We found that XOSs bind to TrCel7A and that the affinity increases commensurate with XOS length. The CBM, on the other hand, did not affect the affinity significantly, which suggests that XOSs may bind to the active site. Activity assays of TrCel7A clearly demonstrated the negative effect of the presence of XOSs on the turnover number. Conclusions On the basis of these binding data and a comparison of XOS inhibition of the activity of the two enzyme variants towards, respectively, soluble and insoluble substrates, we propose a competitive mechanism for XOS inhibition of TrCel7A with phosphoric swollen cellulose as a substrate. PMID:22035059

  2. Test and evaluation document for DOT Specification 7A Type A Packaging. Revision 3

    SciTech Connect

    1996-01-30

    The US Department of Energy (DOE) has been conducting, through several of its operating contractors, an evaluation and testing program to qualify Type A radioactive material packagings per US Department of Transportation (DOT) Specification 7A (DOT-7A) of the Code of Federal Regulations (CFR), Title 49, Part 178 (49 CFR 178). The program is currently administered by the DOE, Office of Facility Safety Analysis, DOE/EH-32, at DOE-Headquarters (DOE-HQ) in Germantown, Maryland. This document summarizes the evaluation and testing performed for all of the packagings successfully qualified in this program.

  3. 13 CFR 120.453 - Responsibilities of PLP Lenders for servicing and liquidating 7(a) loans.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 13 Business Credit and Assistance 1 2010-01-01 2010-01-01 false Responsibilities of PLP Lenders for servicing and liquidating 7(a) loans. 120.453 Section 120.453 Business Credit and Assistance SMALL BUSINESS ADMINISTRATION BUSINESS LOANS Lenders Preferred Lenders Program (plp) § 120.453...

  4. Lmx-1b and Wnt-7a expression in axolotl limb during development and regeneration.

    PubMed

    Shimokawa, Takashi; Yasutaka, Satoru; Kominami, Rieko; Shinohara, Harumichi

    2013-01-01

    Axolotls (Ambystoma mexicanum) have the ability to regenerate amputated limbs throughout their life span. During limb regeneration as well as development, undifferentiated cells in the blastema acquire positional information to reproduce the original pattern along three cardinal limb axes: anteroposterior, proximodistal and dorsoventral. In the present study, we attempted to understand the molecular mechanism involved in patterning of axolotl limb development and regeneration along the dorsoventral (DV) axis. We cloned axolotl Lmx-1b and Wnt-7a, and investigated the expression pattern of these genes in developing and regenerating limbs. In axolotl, unlike in amniotes, Wnt-7a was expressed in a diffuse manner throughout both developing limb bud and regenerating limb blastema. Lmx-1b expression was observed at the dorsal mesenchyme in the developing and regenerating limbs. On the basis of the expression patterns of Lmx-1b and Wnt-7a, it was difficult to identify the interaction between these two genes as reported in amniotes in previous studies. Possibly, with regard to Lmx-1b expression, a Wnt-7a-independent mechanism may exist in axolotl limb development and regeneration.

  5. Semaphorin7A regulates neuroglial plasticity in the adult hypothalamic median eminence.

    PubMed

    Parkash, Jyoti; Messina, Andrea; Langlet, Fanny; Cimino, Irene; Loyens, Anne; Mazur, Danièle; Gallet, Sarah; Balland, Eglantine; Malone, Samuel A; Pralong, François; Cagnoni, Gabriella; Schellino, Roberta; De Marchis, Silvia; Mazzone, Massimiliano; Pasterkamp, R Jeroen; Tamagnone, Luca; Prevot, Vincent; Giacobini, Paolo

    2015-01-01

    Reproductive competence in mammals depends on the projection of gonadotropin-releasing hormone (GnRH) neurons to the hypothalamic median eminence (ME) and the timely release of GnRH into the hypothalamic-pituitary-gonadal axis. In adult rodents, GnRH neurons and the specialized glial cells named tanycytes periodically undergo cytoskeletal plasticity. However, the mechanisms that regulate this plasticity are still largely unknown. We demonstrate that Semaphorin7A, expressed by tanycytes, plays a dual role, inducing the retraction of GnRH terminals and promoting their ensheathment by tanycytic end feet via the receptors PlexinC1 and Itgb1, respectively. Moreover, Semaphorin7A expression is regulated during the oestrous cycle by the fluctuating levels of gonadal steroids. Genetic invalidation of Semaphorin7A receptors in mice induces neuronal and glial rearrangements in the ME and abolishes normal oestrous cyclicity and fertility. These results show a role for Semaphorin7A signalling in mediating periodic neuroglial remodelling in the adult ME during the ovarian cycle.

  6. 13 CFR 120.520 - Purchase of 7(a) loan guarantees.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 13 Business Credit and Assistance 1 2010-01-01 2010-01-01 false Purchase of 7(a) loan guarantees. 120.520 Section 120.520 Business Credit and Assistance SMALL BUSINESS ADMINISTRATION BUSINESS LOANS... has not been cured, provided all business personal property securing the defaulted SBA loan has...

  7. 13 CFR 120.520 - Purchase of 7(a) loan guarantees.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 13 Business Credit and Assistance 1 2012-01-01 2012-01-01 false Purchase of 7(a) loan guarantees. 120.520 Section 120.520 Business Credit and Assistance SMALL BUSINESS ADMINISTRATION BUSINESS LOANS... has not been cured, provided all business personal property securing the defaulted SBA loan has...

  8. 75 FR 3468 - Revised Jurisdictional Thresholds For Section 7A of the Clayton Act

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-01-21

    ... From the Federal Register Online via the Government Publishing Office FEDERAL TRADE COMMISSION Revised Jurisdictional Thresholds For Section 7A of the Clayton Act AGENCY: Federal Trade Commission. ACTION: Notice. SUMMARY: The Federal Trade Commission announces the revised thresholds for the...

  9. Genome Sequence of Streptomyces viridosporus Strain T7A ATCC 39115, a Lignin-Degrading Actinomycete

    SciTech Connect

    Davis, Jennifer R.; Goodwin, Lynne A.; Teshima, Hazuki; Detter, J. Chris; Tapia, Roxanne; Han, Cliff; Huntemann, Marcel; Wei, Chia-Lin; Han, James; Chen, Amy; Kyrpides, Nikos C; Mavromatis, K; Szeto, Ernest; Markowitz, Victor; Ivanova, N; Mikhailova, Natalia; Ovchinnikova, Galina; Pagani, Ioanna; Pati, Amrita; Woyke, Tanja; Pitluck, Sam; Peters, Lin; Nolan, Matt; Land, Miriam L; Sello, Jason K.

    2013-01-01

    We announce the availability of the genome sequence of Streptomyces viridosporus strain T7A ATCC 39115, a plant biomass- degrading actinomycete. This bacterium is of special interest because of its capacity to degrade lignin, an underutilized compo- nent of plants in the context of bioenergy. It has a full complement of genes for plant biomass catabolism.

  10. A novel quantitative trait locus for Fusarium head blight resistance in chromosome 7A of wheat

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A Chinese Spring-Sumai 3 chromosome 7A disomic substitution line (CS-Sumai 3-7ADSL) was reported to have a high level of Fusarium head blight (FHB) resistance for symptom spread within a spike (Type II) and low deoxynivalenol accumulation in infected kernels (Type III), but quantitative trait locus ...

  11. Semaphorin7A regulates neuroglial plasticity in the adult hypothalamic median eminence

    PubMed Central

    Parkash, Jyoti; Messina, Andrea; Langlet, Fanny; Cimino, Irene; Loyens, Anne; Mazur, Danièle; Gallet, Sarah; Balland, Eglantine; Malone, Samuel A.; Pralong, François; Cagnoni, Gabriella; Schellino, Roberta; De Marchis, Silvia; Mazzone, Massimiliano; Pasterkamp, R. Jeroen; Tamagnone, Luca; Prevot, Vincent; Giacobini, Paolo

    2015-01-01

    Reproductive competence in mammals depends on the projection of gonadotropin-releasing hormone (GnRH) neurons to the hypothalamic median eminence (ME) and the timely release of GnRH into the hypothalamic–pituitary–gonadal axis. In adult rodents, GnRH neurons and the specialized glial cells named tanycytes periodically undergo cytoskeletal plasticity. However, the mechanisms that regulate this plasticity are still largely unknown. We demonstrate that Semaphorin7A, expressed by tanycytes, plays a dual role, inducing the retraction of GnRH terminals and promoting their ensheathment by tanycytic end feet via the receptors PlexinC1 and Itgb1, respectively. Moreover, Semaphorin7A expression is regulated during the oestrous cycle by the fluctuating levels of gonadal steroids. Genetic invalidation of Semaphorin7A receptors in mice induces neuronal and glial rearrangements in the ME and abolishes normal oestrous cyclicity and fertility. These results show a role for Semaphorin7A signalling in mediating periodic neuroglial remodelling in the adult ME during the ovarian cycle. PMID:25721933

  12. Cystinuria in a patient with a novel mutation in SLC7A9 gene.

    PubMed

    Koulivand, Leila; Mohammadi, Mehrdad; Ezatpour, Behrouz; Kheirollahi, Majid

    2015-01-01

    Cystinuria, one of the first inborn errors of metabolism, is characterized by hyperexcretion of cystine, arginine, lysine, and ornithine into urine. Cystinuria is genetically classified into types A and B. Mutations in the SLC3A1 gene lead to type A, and type B is caused by mutations in the SLC7A9 gene. We described a 19-year-old woman that had early onset of cystine calculus formation at the age of 3 years. After DNA extraction and polymerase chain reaction, direct sequencing was performed. By these methods, a novel nucleotide substitution c.177G>A in exon 3 of the SLC7A9 gene was found, which had not been reported elsewhere previously. This nucleotide substitution occurs in the extracellular domain of the SLC7A9 gene. In addition, a previously described intron variant c.1136+2/3delT (intron 6 of SLC3A1) in homozygosity status was detected in the patient. To our knowledge, this is the first report of novel nucleotide substitution c.177G>A in exon 3 of the SLC7A9 gene. PMID:25599739

  13. Documentation and analysis for packaging for surface moisture measurement system 7A containers

    SciTech Connect

    Clem, D.K.

    1996-06-17

    This documentation and analysis for packaging documents that two, procured, carbon steel 5-gal drums meet all applicable U.S.Department of Transportation-7A requirements. One container will be used to transport a 0.009 Ci 252 Cf source and the other to transport a 1.7 Ci Am-Be source to and from various 200 Area tank farms.

  14. SLC7A5 act as a potential leukemic transformation target gene in myelodysplastic syndrome

    PubMed Central

    Ma, Yan; Song, Jing; Chen, Bobin; Xu, Xiaoping; Lin, Guowei

    2016-01-01

    Objective Myelodysplastic syndromes (MDS) are a heterogenous group of clonal hematopoietic stem cell disorders characterized by increased risk of leukemic transformation. This study identifies microRNAs(miRNA) and miRNA targets that might represent leukemic transformation markers for MDS. Methods Based on our previously established nested case-control study cohort of MDS patients, we chose paired patients to undergo Angilent 8 × 15K human miRNA microarrays. Target prediction analysis was administrated using targetscan 5.1 software. We further investigated the function of target gene in MDS cell line using siRNA method, including cell proliferation, cell apoptosis, cell cycle and electron microscope. Results Finally we screened a subset of 7 miRNAs to be significantly differentially expressed between the case (at the end of follow up with leukemic transformation) and control group (at the end of follow up without leukemic transformation). Target prediction analysis revealed SLC7A5 was the common target gene of these 7 miRNAs. Further study on the function of SLC7A5 gene in SKM-1 cell line showed that downregulation of SLC7A5 inhibited SKM-1 cells proliferation, increased apoptosis and caused cell cycle arrest in the G0/G1 stage. Conclusion Our data indicate that SLC7A5 gene may act as a potential leukemic transformation target gene in MDS. PMID:26657287

  15. 17 CFR 260.7a-22 - Information unknown or not reasonably available.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 17 Commodity and Securities Exchanges 3 2013-04-01 2013-04-01 false Information unknown or not... Contents § 260.7a-22 Information unknown or not reasonably available. Information required shall be given... which is unknown and is unavailable after reasonable investigation, there shall be included a...

  16. 17 CFR 260.7a-22 - Information unknown or not reasonably available.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 17 Commodity and Securities Exchanges 3 2012-04-01 2012-04-01 false Information unknown or not... Contents § 260.7a-22 Information unknown or not reasonably available. Information required shall be given... which is unknown and is unavailable after reasonable investigation, there shall be included a...

  17. 17 CFR 260.7a-22 - Information unknown or not reasonably available.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 17 Commodity and Securities Exchanges 4 2014-04-01 2014-04-01 false Information unknown or not... Contents § 260.7a-22 Information unknown or not reasonably available. Information required shall be given... which is unknown and is unavailable after reasonable investigation, there shall be included a...

  18. 17 CFR 260.7a-22 - Information unknown or not reasonably available.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 17 Commodity and Securities Exchanges 3 2010-04-01 2010-04-01 false Information unknown or not... Contents § 260.7a-22 Information unknown or not reasonably available. Information required shall be given... which is unknown and is unavailable after reasonable investigation, there shall be included a...

  19. Thyroid hormone induction of human cholesterol 7 alpha-hydroxylase (Cyp7a1) in vitro

    PubMed Central

    Lammel Lindemann, Jan A.; Angajala, Anusha; Engler, David A.; Webb, Paul; Ayers, Stephen D.

    2014-01-01

    Thyroid hormone (TH) modulates serum cholesterol by acting on TH receptor β1 (TRβ1) in liver to regulate metabolic gene sets. In rodents, one important TH regulated step involves induction of Cyp7a1, an enzyme in the cytochrome P450 family, which enhances cholesterol to bile acid conversion and plays a crucial role in regulation of serum cholesterol levels. Current models suggest, however, that Cyp7a1 has lost the capacity to respond to THs in humans. We were prompted to re-examine TH effects on cholesterol metabolic genes in human liver cells by a recent study of a synthetic TH mimetic which showed that serum cholesterol reductions were accompanied by increases in a marker for bile acid synthesis in humans. Here, we show that TH effects upon cholesterol metabolic genes are almost identical in mouse liver, mouse and human liver primary cells and human hepatocyte cell lines. Moreover, Cyp7a1 is a direct TR target gene that responds to physiologic TR levels through a set of distinct response elements in its promoter. These findings suggest that THs regulate cholesterol to bile acid conversion in similar ways in humans and rodent experimental models and that manipulation of hormone signaling pathways could provide a strategy to enhance Cyp7a1 activity in human patients. PMID:24582860

  20. Reduced Let-7a Is Associated with Chemoresistance in Primary Breast Cancer

    PubMed Central

    Deng, Heran; Chen, Kai; Zhu, Liling; Yu, Fengyan; Su, Fengxi

    2015-01-01

    Chemotherapy resistance remains an important problem in the breast cancer clinic. The ability to predict the patients who would respond to a distinct therapy would help to optimize tailored treatment options. miRNAs can mediate a number of genes in response to drug-induced acute cellular stress. Several studies suggest that let-7 miRNA may be involved in the chemosensitivity of cancer cell lines in vitro. However, it is not known whether this phenomenon occurs in clinical breast tumors. The present study showed that lower let-7a expression was associated with epirubicin resistance in primary breast tumors. Moreover, upregulation of let-7a expression sensitized resistant breast tumor cell lines to epirubicin by enhancing cellular apoptosis in vitro. Collectively, these findings indicate that lower expression of let-7a miRNA can induce chemoresistance in breast cancer by enhancing cellular apoptosis and suggest that let-7a may be used as a therapeutic target to modulate epirubicin-based chemotherapy resistance. PMID:26218285

  1. Thrombospondin Type-1 Domain-Containing 7A in Idiopathic Membranous Nephropathy

    PubMed Central

    Meyer-Schwesinger, Catherine; Seitz-Polski, Barbara; Ma, Hong; Zahner, Gunther; Dolla, Guillaume; Hoxha, Elion; Helmchen, Udo; Dabert-Gay, Anne-Sophie; Debayle, Delphine; Merchant, Michael; Klein, Jon; Salant, David J.; Stahl, Rolf A.K.; Lambeau, Gérard

    2014-01-01

    BACKGROUND Idiopathic membranous nephropathy is an autoimmune disease. In approximately 70% of patients, it is associated with autoantibodies against the phospholipase A2 receptor 1 (PLA2R1). Antigenic targets in the remaining patients are unknown. METHODS Using Western blotting, we screened serum samples from patients with idiopathic membranous nephropathy, patients with other glomerular diseases, and healthy controls for antibodies against human native glomerular proteins. We partially purified a putative new antigen, identified this protein by means of mass spectrometry of digested peptides, and validated the results by analysis of recombinant protein expression, immunoprecipitation, and immunohistochemical analysis. RESULTS Serum samples from 6 of 44 patients in a European cohort and 9 of 110 patients in a Boston cohort with anti-PLA2R1–negative idiopathic membranous nephropathy recognized a glomerular protein that was 250 kD in size. None of the serum samples from the 74 patients with idiopathic membranous nephropathy who were sero-positive for anti-PLA2R1 antibodies, from the 76 patients with other glomerular diseases, and from the 44 healthy controls reacted against this antigen. Although this newly identified antigen is clearly different from PLA2R1, it shares some biochemical features, such as N-glycosylation, membranous location, and reactivity with serum only under nonreducing conditions. Mass spectrometry identified this antigen as thrombospondin type-1 domain-containing 7A (THSD7A). All reactive serum samples recognized recombinant THSD7A and immunoprecipitated THSD7A from glomerular lysates. Moreover, immunohistochemical analyses of biopsy samples from patients revealed localization of THSD7A to podocytes, and IgG eluted from one of these samples was specific for THSD7A. CONCLUSIONS In our cohort, 15 of 154 patients with idiopathic membranous nephropathy had circulating autoantibodies to THSD7A but not to PLA2R1, a finding that suggests a distinct

  2. 40 CFR 60.482-7a - Standards: Valves in gas/vapor service and in light liquid service.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...-7a Standards: Valves in gas/vapor service and in light liquid service. (a)(1) Each valve shall be... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Standards: Valves in gas/vapor service and in light liquid service. 60.482-7a Section 60.482-7a Protection of Environment...

  3. 17 CFR 260.7a-16 - Inclusion of items, differentiation between items and answers, omission of instructions.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 17 Commodity and Securities Exchanges 4 2014-04-01 2014-04-01 false Inclusion of items, differentiation between items and answers, omission of instructions. 260.7a-16 Section 260.7a-16 Commodity and... INDENTURE ACT OF 1939 Formal Requirements § 260.7a-16 Inclusion of items, differentiation between items...

  4. 17 CFR 260.7a-16 - Inclusion of items, differentiation between items and answers, omission of instructions.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 17 Commodity and Securities Exchanges 3 2012-04-01 2012-04-01 false Inclusion of items, differentiation between items and answers, omission of instructions. 260.7a-16 Section 260.7a-16 Commodity and... INDENTURE ACT OF 1939 Formal Requirements § 260.7a-16 Inclusion of items, differentiation between items...

  5. 17 CFR 260.7a-16 - Inclusion of items, differentiation between items and answers, omission of instructions.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 17 Commodity and Securities Exchanges 3 2011-04-01 2011-04-01 false Inclusion of items, differentiation between items and answers, omission of instructions. 260.7a-16 Section 260.7a-16 Commodity and... INDENTURE ACT OF 1939 Formal Requirements § 260.7a-16 Inclusion of items, differentiation between items...

  6. 17 CFR 260.7a-16 - Inclusion of items, differentiation between items and answers, omission of instructions.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 17 Commodity and Securities Exchanges 3 2013-04-01 2013-04-01 false Inclusion of items, differentiation between items and answers, omission of instructions. 260.7a-16 Section 260.7a-16 Commodity and... INDENTURE ACT OF 1939 Formal Requirements § 260.7a-16 Inclusion of items, differentiation between items...

  7. 17 CFR 260.7a-16 - Inclusion of items, differentiation between items and answers, omission of instructions.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 17 Commodity and Securities Exchanges 3 2010-04-01 2010-04-01 false Inclusion of items, differentiation between items and answers, omission of instructions. 260.7a-16 Section 260.7a-16 Commodity and... INDENTURE ACT OF 1939 Formal Requirements § 260.7a-16 Inclusion of items, differentiation between items...

  8. Variation in WNT7A is unlikely to be a cause of familial Congenital Talipes Equinovarus

    PubMed Central

    Liu, Guoqing; Inglis, Julie; Cardy, Amanda; Shaw, Duncan; Sahota, Sukhy; Hennekam, Raoul; Sharp, Linda; Miedzybrodzka, Zosia

    2008-01-01

    Background Genetic factors make an important contribution to the aetiology of congenital talipes equinovarus (CTEV), the most common developmental disorder of the lower limb. WNT7A was suggested as a candidate gene for CTEV on the basis of a genome-wide scan for linkage in a large multi-case family. WNT7A is a plausible candidate gene for CTEV as it provides a signal for pattern formation during limb development, and mutation in WNT7A has been reported in a number of limb malformation syndromes. Methods We investigated the role of WNT7A using a family-based linkage approach in our large series of European multi-case CTEV families. Three microsatellite markers were used, of which one (D3S2385) is intragenic, and the other two (D3S2403, D3S1252) are 700 kb 5' to the start and 20 kb from the 3' end of the gene, respectively. Ninety-one CTEV families, comprising 476 individuals of whom 211 were affected, were genotyped. LOD scores using recessive and incomplete-dominant inheritance models, and non-parametric linkage scores, excluded linkage. Results No significant evidence for linkage was observed using either parametric or non-parametric models. LOD scores for the parametric models remained strongly negative in the regions between the markers, and in the 0.5 cM intervals outside the marker map. No significant lod scores were obtained when the data were analysed allowing for heterogeneity. Conclusion Our evidence suggests that the WNT7A gene is unlikely to be a major contributor to the aetiology of familial CTEV. PMID:18538017

  9. Challenges and lessons learned since implementation of the safety pharmacology guidance ICH S7A.

    PubMed

    Valentin, Jean-Pierre; Bass, Alan S; Atrakchi, Aisar; Olejniczak, Klaus; Kannosuke, Fujimori

    2005-01-01

    The International Conference on Harmonization, Topic S7A guidance (ICH S7A) on safety pharmacology for human pharmaceuticals has been in effect for 3 years in Europe, the United States and Japan. Surveys of the pharmaceutical industry, regulatory agencies and the audience attending the 4th Annual Meeting of the Safety Pharmacology Society have helped identify and address areas of controversy, as well as those challenges that have emerged since implementation of the guidance worldwide. Overall, ICH S7A has been successfully implemented. The guidance provides for "Good Laboratory Practice" compliant "safety pharmacology core battery" of studies that are generally performed prior to first administration to humans. The approach is science-driven and specifies the use of robust and sophisticated in vitro and/or in vivo assays. There are, however, some areas that require further refinement/clarification such as the specifics of study design including the selection of dose/concentration, choice of species, modeling of the temporal pharmacodynamic changes in relation to pharmacokinetic profile of parent drug and major metabolites, use of an appropriate sample size, statistical power analysis as a means of demonstrating the sensitivity of the model system, testing of human-specific metabolites and demonstrating not only the model's sensitivity, but also its specificity for predicting adverse events in humans. There was also discussion of when these studies are needed in relation to the clinical development plan. Representatives from the pharmaceutical industry and regulatory agencies see the implementation of ICH S7A as a major step forward towards identifying the risk to Phase 1 and 2 volunteers and patients. It remains to be seen, however, whether and in what ways the ICH S7A-based strategy will contribute to the modification of the integrated risk assessment during the latter stages of clinical development or once drugs have been introduced to the marketplace. PMID

  10. Cystinuria Associated with Different SLC7A9 Gene Variants in the Cat

    PubMed Central

    Raj, Karthik; Osborne, Carl; Giger, Urs

    2016-01-01

    Cystinuria is a classical inborn error of metabolism characterized by a selective proximal renal tubular defect affecting cystine, ornithine, lysine, and arginine (COLA) reabsorption, which can lead to uroliths and urinary obstruction. In humans, dogs and mice, cystinuria is caused by variants in one of two genes, SLC3A1 and SLC7A9, which encode the rBAT and bo,+AT subunits of the bo,+ basic amino acid transporter system, respectively. In this study, exons and flanking regions of the SLC3A1 and SLC7A9 genes were sequenced from genomic DNA of cats (Felis catus) with COLAuria and cystine calculi. Relative to the Felis catus-6.2 reference genome sequence, DNA sequences from these affected cats revealed 3 unique homozygous SLC7A9 missense variants: one in exon 5 (p.Asp236Asn) from a non-purpose-bred medium-haired cat, one in exon 7 (p.Val294Glu) in a Maine Coon and a Sphinx cat, and one in exon 10 (p.Thr392Met) from a non-purpose-bred long-haired cat. A genotyping assay subsequently identified another cystinuric domestic medium-haired cat that was homozygous for the variant originally identified in the purebred cats. These missense variants result in deleterious amino acid substitutions of highly conserved residues in the bo,+AT protein. A limited population survey supported that the variants found were likely causative. The remaining 2 sequenced domestic short-haired cats had a heterozygous variant at a splice donor site in intron 10 and a homozygous single nucleotide variant at a branchpoint in intron 11 of SLC7A9, respectively. This study identifies the first SLC7A9 variants causing feline cystinuria and reveals that, as in humans and dogs, this disease is genetically heterogeneous in cats. PMID:27404572

  11. Cystinuria Associated with Different SLC7A9 Gene Variants in the Cat.

    PubMed

    Mizukami, Keijiro; Raj, Karthik; Osborne, Carl; Giger, Urs

    2016-01-01

    Cystinuria is a classical inborn error of metabolism characterized by a selective proximal renal tubular defect affecting cystine, ornithine, lysine, and arginine (COLA) reabsorption, which can lead to uroliths and urinary obstruction. In humans, dogs and mice, cystinuria is caused by variants in one of two genes, SLC3A1 and SLC7A9, which encode the rBAT and bo,+AT subunits of the bo,+ basic amino acid transporter system, respectively. In this study, exons and flanking regions of the SLC3A1 and SLC7A9 genes were sequenced from genomic DNA of cats (Felis catus) with COLAuria and cystine calculi. Relative to the Felis catus-6.2 reference genome sequence, DNA sequences from these affected cats revealed 3 unique homozygous SLC7A9 missense variants: one in exon 5 (p.Asp236Asn) from a non-purpose-bred medium-haired cat, one in exon 7 (p.Val294Glu) in a Maine Coon and a Sphinx cat, and one in exon 10 (p.Thr392Met) from a non-purpose-bred long-haired cat. A genotyping assay subsequently identified another cystinuric domestic medium-haired cat that was homozygous for the variant originally identified in the purebred cats. These missense variants result in deleterious amino acid substitutions of highly conserved residues in the bo,+AT protein. A limited population survey supported that the variants found were likely causative. The remaining 2 sequenced domestic short-haired cats had a heterozygous variant at a splice donor site in intron 10 and a homozygous single nucleotide variant at a branchpoint in intron 11 of SLC7A9, respectively. This study identifies the first SLC7A9 variants causing feline cystinuria and reveals that, as in humans and dogs, this disease is genetically heterogeneous in cats. PMID:27404572

  12. Cystinuria Associated with Different SLC7A9 Gene Variants in the Cat.

    PubMed

    Mizukami, Keijiro; Raj, Karthik; Osborne, Carl; Giger, Urs

    2016-01-01

    Cystinuria is a classical inborn error of metabolism characterized by a selective proximal renal tubular defect affecting cystine, ornithine, lysine, and arginine (COLA) reabsorption, which can lead to uroliths and urinary obstruction. In humans, dogs and mice, cystinuria is caused by variants in one of two genes, SLC3A1 and SLC7A9, which encode the rBAT and bo,+AT subunits of the bo,+ basic amino acid transporter system, respectively. In this study, exons and flanking regions of the SLC3A1 and SLC7A9 genes were sequenced from genomic DNA of cats (Felis catus) with COLAuria and cystine calculi. Relative to the Felis catus-6.2 reference genome sequence, DNA sequences from these affected cats revealed 3 unique homozygous SLC7A9 missense variants: one in exon 5 (p.Asp236Asn) from a non-purpose-bred medium-haired cat, one in exon 7 (p.Val294Glu) in a Maine Coon and a Sphinx cat, and one in exon 10 (p.Thr392Met) from a non-purpose-bred long-haired cat. A genotyping assay subsequently identified another cystinuric domestic medium-haired cat that was homozygous for the variant originally identified in the purebred cats. These missense variants result in deleterious amino acid substitutions of highly conserved residues in the bo,+AT protein. A limited population survey supported that the variants found were likely causative. The remaining 2 sequenced domestic short-haired cats had a heterozygous variant at a splice donor site in intron 10 and a homozygous single nucleotide variant at a branchpoint in intron 11 of SLC7A9, respectively. This study identifies the first SLC7A9 variants causing feline cystinuria and reveals that, as in humans and dogs, this disease is genetically heterogeneous in cats.

  13. Test and evaluation document for DOT Specification 7A Type A packaging

    SciTech Connect

    Cruse, J.M.

    1992-06-01

    The US Department of Energy (DOE) has been conducting, through several of its operating contractors, an evaluation and testing program to qualify Type A radioactive material packagings per US Department of Transportation Specification 7A (DOT-7A) of the Code of Federal Regulations, Title 49, Part 178, Section 178.350 (49 CFR 178.350). This program is called the DOT-7A Program. The DOT-7A Program is currently administered by the DOE, Division of Quality Verification and Transportation Safety, DOE/EH-33.3, at DOE-Headquarters in Germantown, Maryland. This document presents approximately 200 different packagings that have been determined to meet the requirements for a DOT Specification 7A Type A packaging per 49 CFR 178.350. It was originally prepared in 1987 by Monsanto Research Corporation -- Mound Laboratory for the DOE`s Security Evaluation Program to facilitate the regulation changes implemented by HM-169 for all DOE contractors. In September 1989, the program was transferred to Westinghouse Hanford Company, which is located in Richland, Washington. The specific packaging data contained in this document will serve to meet the requirements of 49 CFR 173.415(a) for ``. . . documentation of tests . . . `` when the packagings are used as prescribed herein. However, shippers are cautioned that additional documentation will be needed to fulfill all of the requirements for a particular shipment. Most important is the evaluation of the contents to be shipped for compatibility with the packaging and that their characteristics are bounded by the simulated contents used in qualification testing.

  14. Thermoascus aurantiacus CBHI/Cel7A Production in Trichoderma reesei on Alternative Carbon Sources

    NASA Astrophysics Data System (ADS)

    Benkő, Zsuzsa; Drahos, Eszter; Szengyel, Zsolt; Puranen, Terhi; Vehmaanperä, Jari; Réczey, Kati

    To develop functional enzymes in cellulose hydrolysis at or above 70°C the cellobiohydrolase (CBHI/Cel7A) of Thermoascus aurantiacus was cloned and expressed in Trichoderma reesei Rut-C30 under the strong cbh1 promoter. Cellulase production of the parental strain and the novel strain (RF6026) was examined in submerged fermentation experiments using various carbon sources, which were lactose, Solka Floc 200 cellulose powder, and steam pretreated corn stover. An industrially feasible production medium was used containing only distiller's spent grain, KH2PO4, and (NH4)2SO4. Enzyme production was followed by measurements of protein concentration, total cellulase enzyme activity (filter paper activity), β-glucosidase activity, CBHI activity, and endogenase I (EGI) activity. The Thermoascus CBHI/Cel7A activity was taken as an indication of the heterologous gene expression under the cbh1 promoter.

  15. A Rotating Stall Type Phenomenon Caused by Cavitation in LE-7A LH2 Turbopump

    NASA Astrophysics Data System (ADS)

    Shimura, Takashi; Yoshida, Makoto; Kamijo, Kenjiro; Uchiumi, Masaharu; Yasutomi, Yoshinobu

    The cause of 350 Hz large-amplitude rotor vibration of the H-2A rocket's LE-7A fuel turbopump was investigated by wideband oscillating-pressure measurement. Measurement was successfully conducted by quartz-type pressure sensors in full-load liquid-hydrogen tests. The phase difference of pressure oscillations between two pressure measurement ports under conditions of pump cavitating operation revealed a phenomenon similar to rotating stall in turbomachinery which had not been previously observed. The rotating speed of a cell was 350 Hz, about a half the rotor speed. This phenomenon is different from rotating cavitation in which the cavitation pattern rotates around the periphery of an impeller faster than shaft rotating speed. Based on these findings, it was concluded that this phenomenon cased the large-amplitude 350 Hz vibration of the LE-7A fuel turbopump. Results of FFT analysis of measured oscillating pressure are presented.

  16. The SLC7A7 Transporter Identifies Microglial Precursors prior to Entry into the Brain.

    PubMed

    Rossi, Federico; Casano, Alessandra Maria; Henke, Katrin; Richter, Kerstin; Peri, Francesca

    2015-05-19

    During development, macrophages invade organs to establish phenotypically and transcriptionally distinct tissue-resident populations. How they invade and colonize these organs is unclear. In particular, it remains to be established whether they arise from naive equivalents that colonize organs randomly or whether there are committed macrophages that follow pre-determined migration paths. Here, by using a combination of genetics and imaging approaches in the zebrafish embryo, we have addressed how macrophages colonize the brain to become microglia. Identification and cloning of a mutant that lacks microglia has shown that Slc7a7, a Leucine/Arginine transporter, defines a restricted macrophage sub-lineage and is necessary for brain colonization. By taking a photoconversion approach, we show that these macrophages give rise to microglia. This study provides direct experimental evidence for the existence of sub-lineages among embryonic macrophages. PMID:25959825

  17. A COL7A1 Mutation Causes Dystrophic Epidermolysis Bullosa in Rotes Höhenvieh Cattle

    PubMed Central

    Menoud, Annie; Welle, Monika; Tetens, Jens; Lichtner, Peter; Drögemüller, Cord

    2012-01-01

    We identified a congenital mechanobullous skin disorder in six calves on a single farm of an endangered German cattle breed in 2010. The condition presented as a large loss of skin distal to the fetlocks and at the mucosa of the muzzle. All affected calves were euthanized on humane grounds due to the severity, extent and progression of the skin and oral lesions. Examination of skin samples under light microscopy revealed detachment of the epidermis from the dermis at the level of the dermo epidermal junction, leading to the diagnosis of a subepidermal bullous dermatosis such as epidermolysis bullosa. The pedigree was consistent with monogenic autosomal recessive inheritance. We localized the causative mutation to an 18 Mb interval on chromosome 22 by homozygosity mapping. The COL7A1 gene encoding collagen type VII alpha 1 is located within this interval and COL7A1 mutations have been shown to cause inherited dystrophic epidermolysis bullosa (DEB) in humans. A SNP in the bovine COL7A1 exon 49 (c.4756C>T) was perfectly associated with the observed disease. The homozygous mutant T/T genotype was exclusively present in affected calves and their parents were heterozygous C/T confirming the assumed recessive mode of inheritance. All known cases and genotyped carriers were related to a single cow, which is supposed to be the founder animal. The mutant T allele was absent in 63 animals from 24 cattle breeds. The identified mutation causes a premature stop codon which leads to a truncated protein representing a complete loss of COL7A1 function (p.R1586*). We thus have identified a candidate causative mutation for this genetic disease using only three cases to unravel its molecular basis. Selection against this mutation can now be used to eliminate the mutant allele from the Rotes Höhenvieh breed. PMID:22715415

  18. In utero copper treatment for Menkes disease associated with a severe ATP7A mutation.

    PubMed

    Haddad, Marie Reine; Macri, Charles J; Holmes, Courtney S; Goldstein, David S; Jacobson, Beryl E; Centeno, Jose A; Popek, Edwina J; Gahl, Willam A; Kaler, Stephen G

    2012-09-01

    Menkes disease is a lethal X-linked recessive neurodegenerative disorder of copper transport caused by mutations in ATP7A, which encodes a copper-transporting ATPase. Early postnatal treatment with copper injections often improves clinical outcomes in affected infants. While Menkes disease newborns appear normal neurologically, analyses of fetal tissues including placenta indicate abnormal copper distribution and suggest a prenatal onset of the metal transport defect. In an affected fetus whose parents found termination unacceptable and who understood the associated risks, we began in utero copper histidine treatment at 31.5 weeks gestational age. Copper histidine (900 μg per dose) was administered directly to the fetus by intramuscular injection (fetal quadriceps or gluteus) under ultrasound guidance. Percutaneous umbilical blood sampling enabled serial measurement of fetal copper and ceruloplasmin levels that were used to guide therapy over a four-week period. Fetal copper levels rose from 17 μg/dL prior to treatment to 45 μg/dL, and ceruloplasmin levels from 39 mg/L to 122 mg/L. After pulmonary maturity was confirmed biochemically, the baby was delivered at 35.5 weeks and daily copper histidine therapy (250 μg sc b.i.d.) was begun. Despite this very early intervention with copper, the infant showed hypotonia, developmental delay, and electroencephalographic abnormalities and died of respiratory failure at 5.5 months of age. The patient's ATP7A mutation (Q724H), which severely disrupted mRNA splicing, resulted in complete absence of ATP7A protein on Western blots. These investigations suggest that prenatally initiated copper replacement is inadequate to correct Menkes disease caused by severe loss-of-function mutations, and that postnatal ATP7A gene addition represents a rational approach in such circumstances.

  19. Genetic epidemiology of gallbladder disease in Mexican Americans and cholesterol 7a-hydroxylase gene variation

    SciTech Connect

    Lin, J.P.; Hanis, C.L.; Boerwinkle, E.

    1994-09-01

    Among Mexican Americans the prevalence of gallbladder disease is markedly elevated. Previous data from both genetic admixture and family studies indicate that there is genetic component to the occurrence of gallbladder disease in Mexican Americans. However, prior to this study no formal genetic analysis of gallbladder disease had been carried out nor had any contributing gene been identified. The results of complex segregation analysis in a sample of 232 Mexican Americans with age- and gender-specific effects influencing the occurrence of gallbladder disease. The estimated frequency of the allele increasing susceptibility was 0.39. The lifetime probabilities that an individual will be affected by gallbladder disease were 1.0, 0.54, and 0.00 for females of genotypes {open_quotes}AA{close_quotes}, {open_quotes}Aa{close_quotes}, and {open_quotes}aa{close_quotes}, respectively, and 0.68, 0.30, and 0.00 for males, respectively. Human cholesterol 7a-hydroxylase is the rate-limiting enzyme in bile acid synthesis. The results of an association study in both a random sample and a matched case/control sample showed that there is a significant association between cholesterol 7a-hydroxylase gene variation and the occurrence of gallbladder disease in Mexican Americans males but not in females. For loci in the 5{prime}-end of the cholesterol 7a-hydroxylase gene, the frequency of the susceptibility alleles was twice as high in gallbladder disease patients compared to controls. The results of a linkage analysis provide evidence that the cholesterol 7a-hydroxylase gene and the inferred gallbladder disease gene are genetically linked.

  20. Structural Insights into the Affinity of Cel7A Carbohydrate-binding Module for Lignin*

    PubMed Central

    Strobel, Kathryn L.; Pfeiffer, Katherine A.; Blanch, Harvey W.; Clark, Douglas S.

    2015-01-01

    The high cost of hydrolytic enzymes impedes the commercial production of lignocellulosic biofuels. High enzyme loadings are required in part due to their non-productive adsorption to lignin, a major component of biomass. Despite numerous studies documenting cellulase adsorption to lignin, few attempts have been made to engineer enzymes to reduce lignin binding. In this work, we used alanine-scanning mutagenesis to elucidate the structural basis for the lignin affinity of Trichoderma reesei Cel7A carbohydrate binding module (CBM). T. reesei Cel7A CBM mutants were produced with a Talaromyces emersonii Cel7A catalytic domain and screened for their binding to cellulose and lignin. Mutation of aromatic and polar residues on the planar face of the CBM greatly decreased binding to both cellulose and lignin, supporting the hypothesis that the cellulose-binding face is also responsible for lignin affinity. Cellulose and lignin affinity of the 31 mutants were highly correlated, although several mutants displayed selective reductions in lignin or cellulose affinity. Four mutants with increased cellulose selectivity (Q2A, H4A, V18A, and P30A) did not exhibit improved hydrolysis of cellulose in the presence of lignin. Further reduction in lignin affinity while maintaining a high level of cellulose affinity is thus necessary to generate an enzyme with improved hydrolysis capability. This work provides insights into the structural underpinnings of lignin affinity, identifies residues amenable to mutation without compromising cellulose affinity, and informs engineering strategies for family one CBMs. PMID:26209638

  1. The Respiratory Chain Supercomplex Organization Is Independent of COX7a2l Isoforms

    PubMed Central

    Mourier, Arnaud; Matic, Stanka; Ruzzenente, Benedetta; Larsson, Nils-Göran; Milenkovic, Dusanka

    2014-01-01

    Summary The organization of individual respiratory chain complexes into supercomplexes or respirasomes has attracted great interest because of the implications for cellular energy conversion. Recently, it was reported that commonly used mouse strains harbor a short COX7a2l (SCAFI) gene isoform that supposedly precludes the formation of complex IV-containing supercomplexes. This claim potentially has serious implications for numerous mouse studies addressing important topics in metabolism, including adaptation to space flights. Using several complementary experimental approaches, we show that mice with the short COX7a2l isoform have normal biogenesis and steady-state levels of complex IV-containing supercomplexes and consequently have normal respiratory chain function. Furthermore, we use a mouse knockout of Lrpprc and show that loss of complex IV compromises respirasome formation. We conclude that the presence of the short COX7a2l isoform in the commonly used C57BL/6 mouse strains does not prevent their use in metabolism research. PMID:25470551

  2. Structural insights into the affinity of Cel7A carbohydrate-binding module for lignin.

    PubMed

    Strobel, Kathryn L; Pfeiffer, Katherine A; Blanch, Harvey W; Clark, Douglas S

    2015-09-11

    The high cost of hydrolytic enzymes impedes the commercial production of lignocellulosic biofuels. High enzyme loadings are required in part due to their non-productive adsorption to lignin, a major component of biomass. Despite numerous studies documenting cellulase adsorption to lignin, few attempts have been made to engineer enzymes to reduce lignin binding. In this work, we used alanine-scanning mutagenesis to elucidate the structural basis for the lignin affinity of Trichoderma reesei Cel7A carbohydrate binding module (CBM). T. reesei Cel7A CBM mutants were produced with a Talaromyces emersonii Cel7A catalytic domain and screened for their binding to cellulose and lignin. Mutation of aromatic and polar residues on the planar face of the CBM greatly decreased binding to both cellulose and lignin, supporting the hypothesis that the cellulose-binding face is also responsible for lignin affinity. Cellulose and lignin affinity of the 31 mutants were highly correlated, although several mutants displayed selective reductions in lignin or cellulose affinity. Four mutants with increased cellulose selectivity (Q2A, H4A, V18A, and P30A) did not exhibit improved hydrolysis of cellulose in the presence of lignin. Further reduction in lignin affinity while maintaining a high level of cellulose affinity is thus necessary to generate an enzyme with improved hydrolysis capability. This work provides insights into the structural underpinnings of lignin affinity, identifies residues amenable to mutation without compromising cellulose affinity, and informs engineering strategies for family one CBMs. PMID:26209638

  3. CDX2 increases SLC7A7 expression and proliferation of pig intestinal epithelial cells

    PubMed Central

    Li, Xiang-guang; Xu, Gao-feng; Zhai, Zhen-ya; Gao, Chun-qi; Yan, Hui-chao; Xi, Qian-yun; Guan, Wu-tai; Wang, Song-bo; Wang, Xiu-qi

    2016-01-01

    Nutrient absorption mediated by nutrient transporters expressed in the intestinal epithelium supplies substrates to support intestinal processes, including epithelial cell proliferation. We evaluated the role of Caudal type homeobox 2 (CDX2), an intestine-specific transcription factor, in the proliferation of pig intestinal epithelial cells (IPEC-1) and searched for novel intestinal nutrient transporter genes activated by CDX2. Our cloned pig CDX2 cDNA contains a “homeobox” DNA binding motif, suggesting it is a transcriptional activator. CDX2 overexpression in IPEC-1 cells increased cell proliferation, the percentage of cells in S/G2 phase, and the abundance of transcripts of the cell cycle-related genes Cyclin A2; Cyclin B; Cyclin D2; proliferating cell nuclear antigen; and cell cycle cyclin-dependent kinases 1, 2 and 4, as well as the predicted CDX2 target genes SLC1A1, SLC5A1 and SLC7A7. In addition, luciferase reporter and chromatin immunoprecipitation assays revealed that CDX2 binds directly to the SLC7A7 promoter. This is the first report of CDX2 function in pig intestinal epithelial cells and identifies SLC7A7 as a novel CDX2 target gene. Our findings show that nutrient transporters are activated during CDX2-induced proliferation of normal intestinal epithelial cells. PMID:27121315

  4. Structural insights into the affinity of Cel7A carbohydrate-binding module for lignin.

    PubMed

    Strobel, Kathryn L; Pfeiffer, Katherine A; Blanch, Harvey W; Clark, Douglas S

    2015-09-11

    The high cost of hydrolytic enzymes impedes the commercial production of lignocellulosic biofuels. High enzyme loadings are required in part due to their non-productive adsorption to lignin, a major component of biomass. Despite numerous studies documenting cellulase adsorption to lignin, few attempts have been made to engineer enzymes to reduce lignin binding. In this work, we used alanine-scanning mutagenesis to elucidate the structural basis for the lignin affinity of Trichoderma reesei Cel7A carbohydrate binding module (CBM). T. reesei Cel7A CBM mutants were produced with a Talaromyces emersonii Cel7A catalytic domain and screened for their binding to cellulose and lignin. Mutation of aromatic and polar residues on the planar face of the CBM greatly decreased binding to both cellulose and lignin, supporting the hypothesis that the cellulose-binding face is also responsible for lignin affinity. Cellulose and lignin affinity of the 31 mutants were highly correlated, although several mutants displayed selective reductions in lignin or cellulose affinity. Four mutants with increased cellulose selectivity (Q2A, H4A, V18A, and P30A) did not exhibit improved hydrolysis of cellulose in the presence of lignin. Further reduction in lignin affinity while maintaining a high level of cellulose affinity is thus necessary to generate an enzyme with improved hydrolysis capability. This work provides insights into the structural underpinnings of lignin affinity, identifies residues amenable to mutation without compromising cellulose affinity, and informs engineering strategies for family one CBMs.

  5. 20 years of ECRH at W7-A and W7-AS

    NASA Astrophysics Data System (ADS)

    Erckmann, V.; Laqua, H. P.; Maaßberg, H.; Marushchenko, N. B.; Kasparek, W.; Müller, G. A.

    2003-11-01

    Basic research on high-power ECRH started 20 years ago at IPP using 28 GHz pulses with 200 kW for 40 ms at the W7-A stellarator. These pilot experiments triggered a strong activity of exploration of the unique capabilities of localized heating and current drive. The achievements in physics were strongly linked with progress in source and transmission line technology. The capability and versatility of ECRH are reviewed using W7-A and W7-AS as example experiments; the latter was shut down on July 31, 2002. The milestone achievements are discussed. Standard heating scenarios such as O-mode and X-mode as well as advanced scenarios like mode-conversion heating via the O-X-B process at different harmonics were investigated and selected results are presented. First experiments with current drive by Bernstein-waves are reported. The physics of wave interaction with stellarator-specific trapped particle populations is discussed. The results from W7-A and W7-AS establish the experimental and technological bases for the 10 MW, CW ECRH system at W7-X, which aims to demonstrate the inherent steady-state capability of stellarators.

  6. Myosin7a Deficiency Results in Reduced Retinal Activity Which Is Improved by Gene Therapy

    PubMed Central

    Colella, Pasqualina; Sommella, Andrea; Marrocco, Elena; Di Vicino, Umberto; Polishchuk, Elena; Garrido, Marina Garcia; Seeliger, Mathias W.; Polishchuk, Roman; Auricchio, Alberto

    2013-01-01

    Mutations in MYO7A cause autosomal recessive Usher syndrome type IB (USH1B), one of the most frequent conditions that combine severe congenital hearing impairment and retinitis pigmentosa. A promising therapeutic strategy for retinitis pigmentosa is gene therapy, however its pre-clinical development is limited by the mild retinal phenotype of the shaker1 (sh1−/−) murine model of USH1B which lacks both retinal functional abnormalities and degeneration. Here we report a significant, early-onset delay of sh1−/− photoreceptor ability to recover from light desensitization as well as a progressive reduction of both b-wave electroretinogram amplitude and light sensitivity, in the absence of significant loss of photoreceptors up to 12 months of age. We additionally show that subretinal delivery to the sh1−/− retina of AAV vectors encoding the large MYO7A protein results in significant improvement of sh1−/− photoreceptor and retinal pigment epithelium ultrastructural anomalies which is associated with improvement of recovery from light desensitization. These findings provide new tools to evaluate the efficacy of experimental therapies for USH1B. In addition, although AAV vectors expressing large genes might have limited clinical applications due to their genome heterogeneity, our data show that AAV-mediated MYO7A gene transfer to the sh1−/− retina is effective. PMID:23991031

  7. A truncated Wnt7a retains full biological activity in skeletal muscle

    NASA Astrophysics Data System (ADS)

    von Maltzahn, Julia; Zinoviev, Radoslav; Chang, Natasha C.; Bentzinger, C. Florian; Rudnicki, Michael A.

    2013-11-01

    Wnt signaling has essential roles during embryonic development and tissue homoeostasis. Wnt proteins are post-translationally modified and the attachment of a palmitate moiety at two conserved residues is believed to be a prerequisite for the secretion and function of Wnt proteins. Here we demonstrate that a mammalian Wnt protein can be fully functional without palmitoylation. We generate a truncated Wnt7a variant, consisting of the C-terminal 137 amino acids lacking the conserved palmitoylation sites and show that it retains full biological activity in skeletal muscle. This includes binding to and signaling through its receptor Fzd7 to stimulate symmetric expansion of satellite stem cells by activating the planar-cell polarity pathway and inducing myofibre hypertrophy by signaling through the AKT/mTOR pathway. Furthermore, this truncated Wnt7a shows enhanced secretion and dispersion compared with the full-length protein. Together, these findings open important new avenues for the development of Wnt7a as a treatment for muscle-wasting diseases and have broad implications for the therapeutic use of Wnts as biologics.

  8. Soluble THSD7A Is an N-Glycoprotein That Promotes Endothelial Cell Migration and Tube Formation in Angiogenesis

    PubMed Central

    Kuo, Meng-Wei; Wang, Chian-Huei; Wu, Hsiao-Chun; Chang, Shing-Jyh; Chuang, Yung-Jen

    2011-01-01

    Background Thrombospondin type I domain containing 7A (THSD7A) is a novel neural protein that is known to affect endothelial migration and vascular patterning during development. To further understand the role of THSD7A in angiogenesis, we investigated the post-translational modification scheme of THS7DA and to reveal the underlying mechanisms by which this protein regulates blood vessel growth. Methodology/Principal Findings Full-length THSD7A was overexpressed in human embryonic kidney 293T (HEK293T) cells and was found to be membrane associated and N-glycosylated. The soluble form of THSD7A, which is released into the cultured medium, was harvested for further angiogenic assays. We found that soluble THSD7A promotes human umbilical vein endothelial cell (HUVEC) migration and tube formation. HUVEC sprouts and zebrafish subintestinal vessel (SIV) angiogenic assays further revealed that soluble THSD7A increases the number of branching points of new vessels. Interestingly, we found that soluble THSD7A increased the formation of filopodia in HUVEC. The distribution patterns of vinculin and phosphorylated focal adhesion kinase (FAK) were also affected, which implies a role for THSD7A in focal adhesion assembly. Moreover, soluble THSD7A increased FAK phosphorylation in HUVEC, suggesting that THSD7A is involved in regulating cytoskeleton reorganization. Conclusions/Significance Taken together, our results indicate that THSD7A is a membrane-associated N-glycoprotein with a soluble form. Soluble THSD7A promotes endothelial cell migration during angiogenesis via a FAK-dependent mechanism and thus may be a novel neuroangiogenic factor. PMID:22194972

  9. 26 CFR 1.482-7A - Methods to determine taxable income in connection with a cost sharing arrangement.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 6 2010-04-01 2010-04-01 false Methods to determine taxable income in connection with a cost sharing arrangement. 1.482-7A Section 1.482-7A Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Regulations Applicable on Or Before January 4, 2009. § 1.482-7A...

  10. Molecular and biochemical analyses of OsRab7, a rice Rab7 homolog.

    PubMed

    Nahm, Min Yeop; Kim, Sam Woong; Yun, Daejin; Lee, Sang Yeol; Cho, Moo Je; Bahk, Jeong Dong

    2003-12-01

    Rab7 is a small GTP-binding protein important in early to late endosome/lysosome vesicular transport in mammalian cells. We have isolated a Rab7 cDNA clone, OsRab7, from a cold-treated rice cDNA library by the subtraction screening method. The cDNA encodes a polypeptide of 206 amino acids with a calculated molecular mass of about 23 kDa. Its predicted amino acid sequence shows significantly high identity with the sequences of other Rab7 proteins. His-tagged OsRab7 bound to radiolabeled GTPgammaS in a specific and stoichiometric manner. Biochemical and structural properties of the Rab7 wild type (WT) protein were compared to those of Q67L and T22N mutants. The detergent 3-([3-cholamidopropyl]dimethylammonio)-1-propane sulfonate (CHAPS) increased the guanine nucleotide binding and hydrolysis activities of Rab7WT. The OsRab7Q67L mutant showed much lower GTPase activity compared to the WT protein untreated with CHAPS, and the T22N mutant showed no GTP binding activity at all. The OsRab7Q67L mutant was constitutively active for guanine nucleotide binding while the T22N mutant (dominant negative) showed no guanine nucleotide binding activity. When bound to GTP, the Rab7WT and the Q67L mutants were protected from tryptic proteolysis. The cleavage pattern of the Rab7T22N mutant, however, was not affected by GTP addition. Northern and Western blot analyses suggested that OsRab7 is distributed in various tissues of rice. Furthermore, expression of a rice Rab7 gene was differentially regulated by various environmental stimuli such as cold, NaCl, dehydration, and ABA. In addition, subcellular localization of OsRab7 was investigated in the Arabidopsis protoplasts by a double-labeling experiment using GFP-fused OsRab7 and FM4-64. GFP-OsRab7 is localized to the vacuolar membrane, suggesting that OsRab7 is implicated in a vesicular transport to the vacuole in plant cells.

  11. Mechanism of product inhibition for cellobiohydrolase Cel7A during hydrolysis of insoluble cellulose.

    PubMed

    Olsen, Johan P; Alasepp, Kadri; Kari, Jeppe; Cruys-Bagger, Nicolaj; Borch, Kim; Westh, Peter

    2016-06-01

    The cellobiohydrolase cellulase Cel7A is extensively utilized in industrial treatment of lignocellulosic biomass under conditions of high product concentrations, and better understanding of inhibition mechanisms appears central in attempts to improve the efficiency of this process. We have implemented an electrochemical biosensor assay for product inhibition studies of cellulases acting on their natural substrate, cellulose. Using this method we measured the hydrolytic rate of Cel7A as a function of both product (inhibitor) concentration and substrate load. This data enabled analyses along the lines of conventional enzyme kinetic theory. We found that the product cellobiose lowered the maximal rate without affecting the Michaelis constant, and this kinetic pattern could be rationalized by two fundamentally distinct molecular mechanisms. One was simple reversibility, that is, an increasing rate of the reverse reaction, lowering the net hydrolytic velocity as product concentrations increase. Strictly this is not a case of inhibition, as no catalytically inactive is formed. The other mechanism that matched the kinetic data was noncompetitive inhibition with an inhibition constant of 490 ± 40 μM. Noncompetitive inhibition implies that the inhibitor binds with comparable strength to either free enzyme or an enzymesubstrate complex, that is, that association between enzyme and substrate has no effect on the binding of the inhibitor. This mechanism is rarely observed, but we argue, that the special architecture of Cel7A with numerous subsites for binding of both substrate and product could give rise to a true noncompetitive inhibition mechanism. Biotechnol. Bioeng. 2016;113: 1178-1186. © 2015 Wiley Periodicals, Inc.

  12. Mechanism of product inhibition for cellobiohydrolase Cel7A during hydrolysis of insoluble cellulose.

    PubMed

    Olsen, Johan P; Alasepp, Kadri; Kari, Jeppe; Cruys-Bagger, Nicolaj; Borch, Kim; Westh, Peter

    2016-06-01

    The cellobiohydrolase cellulase Cel7A is extensively utilized in industrial treatment of lignocellulosic biomass under conditions of high product concentrations, and better understanding of inhibition mechanisms appears central in attempts to improve the efficiency of this process. We have implemented an electrochemical biosensor assay for product inhibition studies of cellulases acting on their natural substrate, cellulose. Using this method we measured the hydrolytic rate of Cel7A as a function of both product (inhibitor) concentration and substrate load. This data enabled analyses along the lines of conventional enzyme kinetic theory. We found that the product cellobiose lowered the maximal rate without affecting the Michaelis constant, and this kinetic pattern could be rationalized by two fundamentally distinct molecular mechanisms. One was simple reversibility, that is, an increasing rate of the reverse reaction, lowering the net hydrolytic velocity as product concentrations increase. Strictly this is not a case of inhibition, as no catalytically inactive is formed. The other mechanism that matched the kinetic data was noncompetitive inhibition with an inhibition constant of 490 ± 40 μM. Noncompetitive inhibition implies that the inhibitor binds with comparable strength to either free enzyme or an enzymesubstrate complex, that is, that association between enzyme and substrate has no effect on the binding of the inhibitor. This mechanism is rarely observed, but we argue, that the special architecture of Cel7A with numerous subsites for binding of both substrate and product could give rise to a true noncompetitive inhibition mechanism. Biotechnol. Bioeng. 2016;113: 1178-1186. © 2015 Wiley Periodicals, Inc. PMID:26636743

  13. WNT7A and PAX6 define corneal epithelium homeostasis and pathogenesis.

    PubMed

    Ouyang, Hong; Xue, Yuanchao; Lin, Ying; Zhang, Xiaohui; Xi, Lei; Patel, Sherrina; Cai, Huimin; Luo, Jing; Zhang, Meixia; Zhang, Ming; Yang, Yang; Li, Gen; Li, Hairi; Jiang, Wei; Yeh, Emily; Lin, Jonathan; Pei, Michelle; Zhu, Jin; Cao, Guiqun; Zhang, Liangfang; Yu, Benjamin; Chen, Shaochen; Fu, Xiang-Dong; Liu, Yizhi; Zhang, Kang

    2014-07-17

    The surface of the cornea consists of a unique type of non-keratinized epithelial cells arranged in an orderly fashion, and this is essential for vision by maintaining transparency for light transmission. Cornea epithelial cells (CECs) undergo continuous renewal from limbal stem or progenitor cells (LSCs), and deficiency in LSCs or corneal epithelium--which turns cornea into a non-transparent, keratinized skin-like epithelium--causes corneal surface disease that leads to blindness in millions of people worldwide. How LSCs are maintained and differentiated into corneal epithelium in healthy individuals and which key molecular events are defective in patients have been largely unknown. Here we report establishment of an in vitro feeder-cell-free LSC expansion and three-dimensional corneal differentiation protocol in which we found that the transcription factors p63 (tumour protein 63) and PAX6 (paired box protein PAX6) act together to specify LSCs, and WNT7A controls corneal epithelium differentiation through PAX6. Loss of WNT7A or PAX6 induces LSCs into skin-like epithelium, a critical defect tightly linked to common human corneal diseases. Notably, transduction of PAX6 in skin epithelial stem cells is sufficient to convert them to LSC-like cells, and upon transplantation onto eyes in a rabbit corneal injury model, these reprogrammed cells are able to replenish CECs and repair damaged corneal surface. These findings suggest a central role of the WNT7A-PAX6 axis in corneal epithelial cell fate determination, and point to a new strategy for treating corneal surface diseases.

  14. Acrolein decreases endothelial cell migration and insulin sensitivity through induction of let-7a.

    PubMed

    O'Toole, Timothy E; Abplanalp, Wesley; Li, Xiaohong; Cooper, Nigel; Conklin, Daniel J; Haberzettl, Petra; Bhatnagar, Aruni

    2014-08-01

    Acrolein is a major reactive component of vehicle exhaust, and cigarette and wood smoke. It is also present in several food substances and is generated endogenously during inflammation and lipid peroxidation. Although previous studies have shown that dietary or inhalation exposure to acrolein results in endothelial activation, platelet activation, and accelerated atherogenesis, the basis for these effects is unknown. Moreover, the effects of acrolein on microRNA (miRNA) have not been studied. Using AGILENT miRNA microarray high-throughput technology, we found that treatment of cultured human umbilical vein endothelial cells with acrolein led to a significant (>1.5-fold) upregulation of 12, and downregulation of 15, miRNAs. Among the miRNAs upregulated were members of the let-7 family and this upregulation was associated with decreased expression of their protein targets, β3 integrin, Cdc34, and K-Ras. Exposure to acrolein attenuated β3 integrin-dependent migration and reduced Akt phosphorylation in response to insulin. These effects of acrolein on endothelial cell migration and insulin signaling were reversed by expression of a let-7a inhibitor. Also, inhalation exposure of mice to acrolein (1 ppm x 6 h/day x 4 days) upregulated let-7a and led to a decrease in insulin-stimulated Akt phosphorylation in the aorta. These results suggest that acrolein exposure has broad effects on endothelial miRNA repertoire and that attenuation of endothelial cell migration and insulin signaling by acrolein is mediated in part by the upregulation of let-7a. This mechanism may be a significant feature of vascular injury caused by inflammation, oxidized lipids, and exposure to environmental pollutants.

  15. Genotypic and phenotypic spectrum of pyridoxine-dependent epilepsy (ALDH7A1 deficiency)

    PubMed Central

    Mills, Philippa B.; Footitt, Emma J.; Mills, Kevin A.; Tuschl, Karin; Aylett, Sarah; Varadkar, Sophia; Hemingway, Cheryl; Marlow, Neil; Rennie, Janet; Baxter, Peter; Dulac, Olivier; Nabbout, Rima; Craigen, William J.; Schmitt, Bernhard; Feillet, François; Christensen, Ernst; De Lonlay, Pascale; Pike, Mike G.; Hughes, M. Imelda; Struys, Eduard A.; Jakobs, Cornelis; Zuberi, Sameer M.

    2010-01-01

    Pyridoxine-dependent epilepsy was recently shown to be due to mutations in the ALDH7A1 gene, which encodes antiquitin, an enzyme that catalyses the nicotinamide adenine dinucleotide-dependent dehydrogenation of l-α-aminoadipic semialdehyde/l-Δ1-piperideine 6-carboxylate. However, whilst this is a highly treatable disorder, there is general uncertainty about when to consider this diagnosis and how to test for it. This study aimed to evaluate the use of measurement of urine l-α-aminoadipic semialdehyde/creatinine ratio and mutation analysis of ALDH7A1 (antiquitin) in investigation of patients with suspected or clinically proven pyridoxine-dependent epilepsy and to characterize further the phenotypic spectrum of antiquitin deficiency. Urinary l-α-aminoadipic semialdehyde concentration was determined by liquid chromatography tandem mass spectrometry. When this was above the normal range, DNA sequencing of the ALDH7A1 gene was performed. Clinicians were asked to complete questionnaires on clinical, biochemical, magnetic resonance imaging and electroencephalography features of patients. The clinical spectrum of antiquitin deficiency extended from ventriculomegaly detected on foetal ultrasound, through abnormal foetal movements and a multisystem neonatal disorder, to the onset of seizures and autistic features after the first year of life. Our relatively large series suggested that clinical diagnosis of pyridoxine dependent epilepsy can be challenging because: (i) there may be some response to antiepileptic drugs; (ii) in infants with multisystem pathology, the response to pyridoxine may not be instant and obvious; and (iii) structural brain abnormalities may co-exist and be considered sufficient cause of epilepsy, whereas the fits may be a consequence of antiquitin deficiency and are then responsive to pyridoxine. These findings support the use of biochemical and DNA tests for antiquitin deficiency and a clinical trial of pyridoxine in infants and children with

  16. The brown and brite adipocyte marker Cox7a1 is not required for non-shivering thermogenesis in mice

    PubMed Central

    Maurer, Stefanie F.; Fromme, Tobias; Grossman, Lawrence I.; Hüttemann, Maik; Klingenspor, Martin

    2015-01-01

    The cytochrome c oxidase subunit isoform Cox7a1 is highly abundant in skeletal muscle and heart and influences enzyme activity in these tissues characterised by high oxidative capacity. We identified Cox7a1, well-known as brown adipocyte marker gene, as a cold-responsive protein of brown adipose tissue. We hypothesised a mechanistic relationship between cytochrome c oxidase activity and Cox7a1 protein levels affecting the oxidative capacity of brown adipose tissue and thus non-shivering thermogenesis. We subjected wildtype and Cox7a1 knockout mice to different temperature regimens and tested characteristics of brown adipose tissue activation. Cytochrome c oxidase activity, uncoupling protein 1 expression and maximal norepinephrine-induced heat production were gradually increased during cold-acclimation, but unaffected by Cox7a1 knockout. Moreover, the abundance of uncoupling protein 1 competent brite cells in white adipose tissue was not influenced by presence or absence of Cox7a1. Skin temperature in the interscapular region of neonates was lower in uncoupling protein 1 knockout pups employed as a positive control, but not in Cox7a1 knockout pups. Body mass gain and glucose tolerance did not differ between wildtype and Cox7a1 knockout mice fed with high fat or control diet. We conclude that brown adipose tissue function in mice does not require the presence of Cox7a1. PMID:26635001

  17. Cloning, expression, purification and crystallization of Schizosaccharomyces pombe Set7, a putative histone methyltransferase.

    PubMed

    Mevius, Damiaan E H F; Shen, Yunpeng; Morishita, Masayo; di Luccio, Eric

    2016-04-01

    Dysfunction of histone-modifying enzymes affects chromatin regulation and is involved in carcinogenesis, tumour progression and other diseases. Histone methyltransferases are a family of key histone-modifying enzymes, but their structures, functions and mechanisms are incompletely understood, thus constraining drug-design efforts. Here, preliminary steps towards structure-function studies of Schizosaccharomyces pombe Set7, a putative histone methyltransferase and the first yeast full-length SET-domain-containing protein to be studied using X-ray crystallography, are reported. The methods from cloning to X-ray diffraction and phasing are discussed and the results will aid in prospective studies of histone-modifying enzymes.

  18. Shuttle Flight-7A DTO-261: Pre and Post Flight Analysis

    NASA Technical Reports Server (NTRS)

    Kaouk, Mohamed; McNeill, Scot; Haley, Sydney; Grygier, Michael; Bartkowicz, Theodore; Rachal, P. Brian; Peart, Walter

    2004-01-01

    This paper reports on the analysis to validate and correlate the analytical models of ISS-Shuttle Flight-7A configuration. On-orbit dynamic responses were measured during an intentional dynamic test (DTO-261) by the Shuttle video camera photogrammetric system and the Internal Wireless Instrumentation System (IWIS). Modal analyses were performed on the measured data to extract modal parameters including frequency, damping, and mode shapes. Correlation and comparisons between test and analytical modal parameters were performed to assess the accuracy of the models.

  19. Expression of WNT genes in cervical cancer-derived cells: Implication of WNT7A in cell proliferation and migration

    SciTech Connect

    Ramos-Solano, Moisés; Meza-Canales, Ivan D.; Torres-Reyes, Luis A.; Alvarez-Zavala, Monserrat; and others

    2015-07-01

    According to the multifactorial model of cervical cancer (CC) causation, it is now recognized that other modifications, in addition to Human papillomavirus (HPV) infection, are necessary for the development of this neoplasia. Among these, it has been proposed that a dysregulation of the WNT pathway might favor malignant progression of HPV-immortalized keratinocytes. The aim of this study was to identify components of the WNT pathway differentially expressed in CC vs. non-tumorigenic, but immortalized human keratinocytes. Interestingly, WNT7A expression was found strongly downregulated in cell lines and biopsies derived from CC. Restoration of WNT7A in CC-derived cell lines using a lentiviral gene delivery system or after adding a recombinant human protein decreases cell proliferation. Likewise, WNT7A silencing in non-tumorigenic cells markedly accelerates proliferation. Decreased WNT7A expression was due to hypermethylation at particular CpG sites. To our knowledge, this is the first study reporting reduced WNT7A levels in CC-derived cells and that ectopic WNT7A restoration negatively affects cell proliferation and migration. - Highlights: • WNT7A is expressed in normal keratinocytes or cervical cells without lesion. • WNT7A is significantly reduced in cervical cancer-derived cells. • Restoration of WNT7A expression in HeLa decreases proliferation and cell migration. • Silencing of WNT7A in HaCaT induces an increased proliferation and migration rate. • Decreased WNT7A expression in this model is due to hypermethylation.

  20. Crystal structure of cytochrome c3 from Desulfovibrio desulfuricans Norway at 1.7 A resolution.

    PubMed

    Czjzek, M; Payan, F; Guerlesquin, F; Bruschi, M; Haser, R

    1994-11-01

    The crystal structure of cytochrome c3 (M(r) 13,000) from Desulfovibrio desulfuricans (118 residues, four heme groups) has been crystallographically refined to 1.7 A resolution using a simulated annealing method, based on the structure-model at 2.5 A resolution, already published. The final R-factor for 10,549 reflections was 0.198 covering the range from 5.5 to 1.7 A resolution. The individual temperature factors were refined for a total of 1059 protein atoms, together with 126 bound solvent molecules. The structure has been analyzed with respect to its detailed conformational properties, secondary structure features, temperature factor behaviour, bound solvent sites and heme geometry and ligation. The characteristic secondary structures of the polypeptide chain of this molecule are one extended alpha-helix, a short beta-strand and 13 reverse turns. The four heme groups are located in different structural environments, all highly exposed to solvent. The particular structural features of the heme environments are compared to the four hemes of the cytochrome c3 from Desulfovibrio vulgaris Miyazaki. PMID:7966289

  1. TEST & EVALUATION REPORT FOR THE HEDGEHOG-II PACKAGING SYSTEMS DOT-7A TYPE A CONTAINER

    SciTech Connect

    KELLY, D.L.

    2003-12-29

    This report documents the US. Department of Transportation Specification 7A (DOT-7A) Type A compliance test and evaluation results for the Hedgehog-II packaging systems. The approved Hedgehog-II packaging configurations provide primary and secondary containment. The approved packaging configurations described within this report are designed to ship Type A quantities of radioactive materials, normal form. Contents may be in solid or liquid form. Liquids transported in the approved 1 L glass bottle assembly shall have a specific gravity of less than or equal to 1.6. Liquids transported in all other approved configurations shall have a specific gravity of less than or equal to 2.0. The solid contents, including packaging, are limited in weight to the gross weight of the as-tested liquids and bottles. The approved Hedgehog-II packaging configurations described in this report may be transported by air, and have been evaluated as meeting the applicable International Air Transport Association/International Civil Aviation Organization (IATA/ICAO) Dangerous Goods Regulations in addition to the DOT requirements.

  2. Temperature Effects on Kinetic Parameters and Substrate Affinity of Cel7A Cellobiohydrolases*

    PubMed Central

    Sørensen, Trine Holst; Cruys-Bagger, Nicolaj; Windahl, Michael Skovbo; Badino, Silke Flindt; Borch, Kim; Westh, Peter

    2015-01-01

    We measured hydrolytic rates of four purified cellulases in small increments of temperature (10–50 °C) and substrate loads (0–100 g/liter) and analyzed the data by a steady state kinetic model that accounts for the processive mechanism. We used wild type cellobiohydrolases (Cel7A) from mesophilic Hypocrea jecorina and thermophilic Rasamsonia emersonii and two variants of these enzymes designed to elucidate the role of the carbohydrate binding module (CBM). We consistently found that the maximal rate increased strongly with temperature, whereas the affinity for the insoluble substrate decreased, and as a result, the effect of temperature depended strongly on the substrate load. Thus, temperature had little or no effect on the hydrolytic rate in dilute substrate suspensions, whereas strong temperature activation (Q10 values up to 2.6) was observed at saturating substrate loads. The CBM had a dual effect on the activity. On one hand, it diminished the tendency of heat-induced desorption, but on the other hand, it had a pronounced negative effect on the maximal rate, which was 2-fold larger in variants without CBM throughout the investigated temperature range. We conclude that although the CBM is beneficial for affinity it slows down the catalytic process. Cel7A from the thermophilic organism was moderately more activated by temperature than the mesophilic analog. This is in accord with general theories on enzyme temperature adaptation and possibly relevant information for the selection of technical cellulases. PMID:26183777

  3. Temperature Effects on Kinetic Parameters and Substrate Affinity of Cel7A Cellobiohydrolases.

    PubMed

    Sørensen, Trine Holst; Cruys-Bagger, Nicolaj; Windahl, Michael Skovbo; Badino, Silke Flindt; Borch, Kim; Westh, Peter

    2015-09-01

    We measured hydrolytic rates of four purified cellulases in small increments of temperature (10-50 °C) and substrate loads (0-100 g/liter) and analyzed the data by a steady state kinetic model that accounts for the processive mechanism. We used wild type cellobiohydrolases (Cel7A) from mesophilic Hypocrea jecorina and thermophilic Rasamsonia emersonii and two variants of these enzymes designed to elucidate the role of the carbohydrate binding module (CBM). We consistently found that the maximal rate increased strongly with temperature, whereas the affinity for the insoluble substrate decreased, and as a result, the effect of temperature depended strongly on the substrate load. Thus, temperature had little or no effect on the hydrolytic rate in dilute substrate suspensions, whereas strong temperature activation (Q10 values up to 2.6) was observed at saturating substrate loads. The CBM had a dual effect on the activity. On one hand, it diminished the tendency of heat-induced desorption, but on the other hand, it had a pronounced negative effect on the maximal rate, which was 2-fold larger in variants without CBM throughout the investigated temperature range. We conclude that although the CBM is beneficial for affinity it slows down the catalytic process. Cel7A from the thermophilic organism was moderately more activated by temperature than the mesophilic analog. This is in accord with general theories on enzyme temperature adaptation and possibly relevant information for the selection of technical cellulases. PMID:26183777

  4. Binding and movement of individual Cel7A cellobiohydrolases on crystalline cellulose surfaces revealed by single-molecule fluorescence imaging.

    PubMed

    Jung, Jaemyeong; Sethi, Anurag; Gaiotto, Tiziano; Han, Jason J; Jeoh, Tina; Gnanakaran, Sandrasegaram; Goodwin, Peter M

    2013-08-16

    The efficient catalytic conversion of biomass to bioenergy would meet a large portion of energy requirements in the near future. A crucial step in this process is the enzyme-catalyzed hydrolysis of cellulose to glucose that is then converted into fuel such as ethanol by fermentation. Here we use single-molecule fluorescence imaging to directly monitor the movement of individual Cel7A cellobiohydrolases from Trichoderma reesei (TrCel7A) on the surface of insoluble cellulose fibrils to elucidate molecular level details of cellulase activity. The motion of multiple, individual TrCel7A cellobiohydrolases was simultaneously recorded with ∼15-nm spatial resolution. Time-resolved localization microscopy provides insights on the activity of TrCel7A on cellulose and informs on nonproductive binding and diffusion. We measured single-molecule residency time distributions of TrCel7A bound to cellulose both in the presence of and absence of cellobiose the major product and a potent inhibitor of Cel7A activity. Combining these results with a kinetic model of TrCel7A binding provides microscopic insight into interactions between TrCel7A and the cellulose substrate. PMID:23818525

  5. Hypoxia-inducible miR-210 contributes to preeclampsia via targeting thrombospondin type I domain containing 7A.

    PubMed

    Luo, Rongcan; Wang, Yongqing; Xu, Peng; Cao, Guangming; Zhao, Yangyu; Shao, Xuan; Li, Yu-xia; Chang, Cheng; Peng, Chun; Wang, Yan-ling

    2016-01-22

    Preeclampsia, a relatively common pregnancy disorder, is a major contributor to maternal mortality and morbidity worldwide. An elevation in microRNA-210 (miR-210) expression in the placenta has been reported to be associated with preeclampsia. Our bioinformatic analysis showed that thrombospondin type I domain containing 7A (THSD7A) is a predicted target for miR-210. The aim of this study was to determine whether miR-210 is involved in preeclampsia through its targeting of THSD7A in human placental trophoblasts. In preeclamptic placental tissues, THSD7A levels were significantly downregulated, and were inversely correlated with the levels of miR-210. THSD7A was validated as a direct target of miR-210 using quantitative real time PCR (qRT-PCR), Western blotting, and dual luciferase assays in HTR8/SVneo cells. Transwell insert invasion assays showed that THSD7A mediated the invasion-inhibitory effect of miR-210 in HTR8/SVneo cells. Interestingly, hypoxia markedly increased miR-210 expression while suppressing THSD7A expression in a time-dependent manner in HTR8/SVneo cells. This study provides novel data on the function of THSD7A in human placental cells, and extends our knowledge of how miR-210 is involved in the development of the preeclampsia.

  6. Hypoxia-inducible miR-210 contributes to preeclampsia via targeting thrombospondin type I domain containing 7A

    PubMed Central

    Luo, Rongcan; Wang, Yongqing; Xu, Peng; Cao, Guangming; Zhao, Yangyu; Shao, Xuan; Li, Yu-xia; Chang, Cheng; Peng, Chun; Wang, Yan-ling

    2016-01-01

    Preeclampsia, a relatively common pregnancy disorder, is a major contributor to maternal mortality and morbidity worldwide. An elevation in microRNA-210 (miR-210) expression in the placenta has been reported to be associated with preeclampsia. Our bioinformatic analysis showed that thrombospondin type I domain containing 7A (THSD7A) is a predicted target for miR-210. The aim of this study was to determine whether miR-210 is involved in preeclampsia through its targeting of THSD7A in human placental trophoblasts. In preeclamptic placental tissues, THSD7A levels were significantly downregulated, and were inversely correlated with the levels of miR-210. THSD7A was validated as a direct target of miR-210 using quantitative real time PCR (qRT-PCR), Western blotting, and dual luciferase assays in HTR8/SVneo cells. Transwell insert invasion assays showed that THSD7A mediated the invasion-inhibitory effect of miR-210 in HTR8/SVneo cells. Interestingly, hypoxia markedly increased miR-210 expression while suppressing THSD7A expression in a time-dependent manner in HTR8/SVneo cells. This study provides novel data on the function of THSD7A in human placental cells, and extends our knowledge of how miR-210 is involved in the development of the preeclampsia. PMID:26796133

  7. Test Plan for Westinghouse Hanford Company`s Hedgehog Shielded Container, Docket 94-39-7A, Type A Container

    SciTech Connect

    Kelly, D.L.

    1995-02-27

    This report documents the US Department of Transportation Specification 7A Type A (DOT-7A) compliance testing to be followed for qualification of the Westinghouse Hanford Company`s Hedgehog Shielded Container for use as a Type A packaging. The packaging configurations being tested are intended for liquids and solids, and for air transportation.

  8. 26 CFR 1.482-7A - Methods to determine taxable income in connection with a cost sharing arrangement.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 6 2012-04-01 2012-04-01 false Methods to determine taxable income in connection with a cost sharing arrangement. 1.482-7A Section 1.482-7A Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Regulations Applicable on Or Before January 4, 2009....

  9. 26 CFR 1.482-7A - Methods to determine taxable income in connection with a cost sharing arrangement.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 6 2013-04-01 2013-04-01 false Methods to determine taxable income in connection with a cost sharing arrangement. 1.482-7A Section 1.482-7A Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Regulations Applicable on Or Before January 4, 2009....

  10. 26 CFR 1.482-7A - Methods to determine taxable income in connection with a cost sharing arrangement.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 6 2014-04-01 2014-04-01 false Methods to determine taxable income in connection with a cost sharing arrangement. 1.482-7A Section 1.482-7A Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Regulations Applicable on Or Before January 4, 2009....

  11. 26 CFR 1.482-7A - Methods to determine taxable income in connection with a cost sharing arrangement.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 6 2011-04-01 2011-04-01 false Methods to determine taxable income in connection with a cost sharing arrangement. 1.482-7A Section 1.482-7A Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Regulations Applicable on Or Before January 4, 2009....

  12. 19 CFR 4.7c - Vessel stow plan.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... the time prescribed in paragraph (a) of this section via the CBP-approved electronic data interchange... paragraph (d)(1) of this section in the event that any necessary modifications to the approved electronic data interchange system are not yet in place or for any other reason. Notice of any such delay will...

  13. Synthesis and properties of lignin peroxidase from Streptomyces viridosporus T7A

    SciTech Connect

    Lodha, S.J.; Korus, R.A.; Crawford, D.L.

    1991-12-31

    The production of lignin peroxidase by Streptomyces viridosporus T7A was studied in shake flasks and under aerobic conditions in a 7.5-L batch fermentor. Lignin peroxidase synthesis was found to be strongly affected by catabolite repression. Lignin peroxidase was a non-growth-associated, secondary metabolite. The maximum lignin peroxidase activity was 0.064 U/mL at 36 h. In order to maximize lignin peroxidase activity, optimal conditions were determined. The optimal incubation temperature, pH, and substrate (2,4-dichlorophenol) concentration for the enzyme assays were 45{degrees}C, 6, and 3 m-M, respectively. Stability of lignin peroxidase was determined at 37, 45, and 60{degrees}C, and over the pH range 4-9.

  14. Soft x-ray beamline BL7A at the UVSOR

    SciTech Connect

    Murata, T. ); Matsukawa, T. ); Naoe, S. ); Horigome, T.; Matsudo, O.; Watanabe, M. )

    1992-01-01

    A vacuum-compatible double-crystal monochromator with constant exit-beam height has been installed and operated for several years at beamline BL7A at the UVSOR facility at the Institute for Molecular Science, Okazaki, Japan, The beamline is used both for the photons from a normal bending-magnet section and those from a 4 T superconducting three-pole horizontal wiggler. Various pairs of monochromator crystals are being used with sufficient intensity and signal to noise ratio of the output signal. Basic structure of the beamline, the mechanism of the monochromator, and some typical spectra of materials with absorption edges between 850 eV and 4 keV are reported.

  15. Unsteady blade pressure measurements for the SR-7A propeller at cruise conditions

    NASA Technical Reports Server (NTRS)

    Heidelberg, L. J.; Nallasamy, M.

    1990-01-01

    The unsteady blade surface pressures were measured on the SR-7A propeller. The freestream Mach number, inflow angle, and advance ratio were varied while measurements were made at nine blade stations. At a freestream Mach number of 0.8, the data in terms of unsteady pressure coefficient versus azimuth angle are compared to an unsteady three-dimensional Euler solution, yielding very encouraging results. The code predicts the shape (phase) of the waveform very well, while the magnitude is over-predicted in many cases. At tunnel Mach numbers below 0.6, an unusually large response on the suction surface at 0.15 chord and 0.88 radius was observed. The behavior of this response suggests the presence of a leading-edge vortex.

  16. Unsteady blade pressure measurements for the SR-7A propeller at cruise conditions

    NASA Technical Reports Server (NTRS)

    Heidelberg, L. J.; Nallasamy, M.

    1990-01-01

    The unsteady blade surface pressures were measured on the SR-7A propeller. The freestream Mach no., inflow angle, and advance ratio were varied while measurements were made at nine blade stations. At a freestream Mach no. of 0.8, the data in terms of unsteady pressure coefficient vs. azimuth angle are compared to an unsteady 3-D Euler solution, yielding very encouraging results. The code predicts the shape (phase) of the waveform very well, while the magnitude is over-predicted in many cases. At tunnel Mach nos. below 0.6, an unusually large response on the suction surface at 0.15 chord and 0.88 radius was observed. The behavior of this response suggests the presence of a leading edge vortex. The midchord measuring stations on the suction surface exhibit a response that leads the forcing function while most other locations show a phase lag.

  17. Structural insights into human Kif7, a kinesin involved in Hedgehog signalling

    PubMed Central

    Klejnot, Marta; Kozielski, Frank

    2012-01-01

    Kif7, a member of the kinesin 4 superfamily, is implicated in a variety of diseases including Joubert, hydrolethalus and acrocallosal syndromes. It is also involved in primary cilium formation and the Hedgehog signalling pathway and may play a role in cancer. Its activity is crucial for embryonic development. Kif7 and Kif27, a closely related kinesin in the same subfamily, are orthologues of the Drosophila melano­gaster kinesin-like protein Costal-2 (Cos2). In vertebrates, they work together to fulfil the role of the single Cos2 gene in Drosophila. Here, the high-resolution structure of the human Kif7 motor domain is reported and is compared with that of conventional kinesin, the founding member of the kinesin superfamily. These data are a first step towards structural characterization of a kinesin-4 family member and of this interesting molecular motor of medical significance. PMID:22281744

  18. Molecular simulation evidence for processive motion of Trichoderma reesei Cel7A during cellulose depolymerization

    NASA Astrophysics Data System (ADS)

    Zhao, Xiongce; Rignall, Tauna R.; McCabe, Clare; Adney, William S.; Himmel, Michael E.

    2008-07-01

    We present free energy calculations for the Trichoderma reesei Cel7A (cellobiohydrolase I) linker peptide from molecular dynamics simulations directed towards understanding the linker role in cellulose hydrolysis. The calculations predict an energy storage mechanism of the linker under stretching/compression that is consistent with processive depolymerization. The linker exhibits two stable states at lengths of 2.5 nm and 5.5 nm during extension/compression, with a free energy difference of 10.5 kcal/mol between the two states separated by an energy barrier. The switching between stable states supports the hypothesis that the linker peptide has the capacity to store energy in a manner similar to a spring.

  19. Detailed noise measurements on the SR-7A propeller: Tone behavior with helical tip Mach number

    NASA Technical Reports Server (NTRS)

    Dittmar, James H.; Hall, David G.

    1991-01-01

    Detailed noise measurements were taken on the SR-7A propeller to investigate the behavior of the noise with helical tip Mach number and then to level off as Mach number was increased further. This behavior was further investigated by obtaining detailed pressure-time histories of data. The pressure-time histories indicate that a portion of the primary pressure pulse is progressively cancelled by a secondary pulse which results in the noise leveling off as the helical tip Mach number is increased. This second pulse appears to originate on the same blade as the primary pulse and is in some way connected to the blade itself. This leaves open the possibility of redesigning the blade to improve the cancellation; thereby, the propeller noise is reduced.

  20. A new experiment station on beamline 4B7A at Beijing Synchrotron Radiation Facility

    NASA Astrophysics Data System (ADS)

    Zheng, L.; Zhao, Y. D.; Tang, K.; Ma, C. Y.; Hong, C. H.; Han, Y.; Cui, M. Q.; Guo, Z. Y.

    2014-11-01

    A new experiment station was installed on beamline 4B7A at Beijing Synchrotron Radiation Facility (BSRF), making it possible to record X-ray absorption fine structure (XAFS) spectrum in three modes over an energy range from 1750 eV to 6000 eV. A 13-element Si(Li) array detector and a single-element SDD detector were used to acquire data in partial fluorescence yield (PFY) mode. Two low-pressure noble gas ion chambers were adopted for measuring XAFS in transmission mode. In total electron yield (TEY) mode the current of sample is recorded. Solid, wet and liquid samples were suitable for this experimental station. Some representative results obtained from this station were shown and discussed.

  1. X-ray crystallographic characterization of new soluble endohedral fullerenes utilizing the popular C82 bucky cage. Isolation and structural characterization of Sm@C3v(7)-C82, Sm@C(s)(6)-C82, and Sm@C2(5)-C82.

    PubMed

    Yang, Hua; Jin, Hongxiao; Wang, Xinqing; Liu, Ziyang; Yu, Meilan; Zhao, Fukun; Mercado, Brandon Q; Olmstead, Marilyn M; Balch, Alan L

    2012-08-29

    Three isomers of Sm@C(82) that are soluble in organic solvents were obtained from the carbon soot produced by vaporization of hollow carbon rods doped with Sm(2)O(3)/graphite powder in an electric arc. These isomers were numbered as Sm@C(82)(I), Sm@C(82)(II), and Sm@C(82)(III) in order of their elution times from HPLC chromatography on a Buckyprep column with toluene as the eluent. The identities of isomers, Sm@C(82)(I) as Sm@C(s)(6)-C(82), Sm@C(82)(II) as Sm@C(3v)(7)-C(82), and Sm@C(82)(III) as Sm@C(2)(5)-C(82), were determined by single-crystal X-ray diffraction on cocrystals formed with Ni(octaethylporphyrin). For endohedral fullerenes like La@C(82), which have three electrons transferred to the cage to produce the M(3+)@(C(82))(3-) electronic distribution, generally only two soluble isomers (e.g., La@C(2v)(9)-C(82) (major) and La@C(s)(6)-C(82) (minor)) are observed. In contrast, with samarium, which generates the M(2+)@(C(82))(2-) electronic distribution, five soluble isomers of Sm@C(82) have been detected, three in this study, the other two in two related prior studies. The structures of the four Sm@C(82) isomers that are currently established are Sm@C(2)(5)-C(82), Sm@C(s)(6)-C(82), Sm@C(3v)(7)-C(82), and Sm@C(2v)(9)-C(82). All of these isomers obey the isolated pentagon rule (IPR) and are sequentially interconvertable through Stone-Wales transformations. PMID:22860880

  2. VERIFICATION OF THE DEFENSE WASTE PROCESSING FACILITY'S (DWPF) PROCESS DIGESTION METHOD FOR THE SLUDGE BATCH 7A QUALIFICATION SAMPLE

    SciTech Connect

    Click, D.; Edwards, T.; Jones, M.; Wiedenman, B.

    2011-03-14

    For each sludge batch that is processed in the Defense Waste Processing Facility (DWPF), the Savannah River National Laboratory (SRNL) performs confirmation of the applicability of the digestion method to be used by the DWPF lab for elemental analysis of Sludge Receipt and Adjustment Tank (SRAT) receipt samples and SRAT product process control samples. DWPF SRAT samples are typically dissolved using a room temperature HF-HNO{sub 3} acid dissolution (i.e., DWPF Cold Chem Method, see DWPF Procedure SW4-15.201) and then analyzed by inductively coupled plasma - atomic emission spectroscopy (ICP-AES). This report contains the results and comparison of data generated from performing the Aqua Regia (AR), Sodium peroxide/Hydroxide Fusion (PF) and DWPF Cold Chem (CC) method digestions of Sludge Batch 7a (SB7a) SRAT Receipt and SB7a SRAT Product samples. The SB7a SRAT Receipt and SB7a SRAT Product samples were prepared in the SRNL Shielded Cells, and the SRAT Receipt material is representative of the sludge that constituates the SB7a Batch or qualification composition. This is the sludge in Tank 51 that is to be transferred into Tank 40, which will contain the heel of Sludge Batch 6 (SB6), to form the Sb7a Blend composition.

  3. Membranous Nephropathy with an Enhanced Granular Expression of Thrombospondin Type-1 Domain-containing 7A in a Pregnant Woman.

    PubMed

    Iwakura, Takamasa; Fujigaki, Yoshihide; Katahashi, Naoko; Sato, Taichi; Ishigaki, Sayaka; Tsuji, Naoko; Naito, Yoshitaka; Isobe, Shinsuke; Ono, Masashi; Sakao, Yukitoshi; Tsuji, Takayuki; Ohashi, Naro; Kato, Akihiko; Miyajima, Hiroaki; Yasuda, Hideo

    2016-01-01

    A 30-year-old woman with proteinuria first noted at 26 weeks of gestation was admitted to undergo further evaluation. A renal biopsy revealed membranous nephropathy (MN). There was no evidence of any secondary MN. Prednisolone was initiated 6 months after delivery. Four months later, her urine protein became negative. Enhanced granular staining for thrombospondin type-1 domain-containing 7A (THSD7A) in the glomeruli was retrospectively detected in a biopsy specimen. A literature review revealed that 60% of cases of THSD7A-related MN occurred in women of childbearing age. Therefore, THSD7A-related MN should be considered in female patients presenting with idiopathic MN in childbearing age.

  4. Synthesis and reactivity of N-heterocycle-B(C6F5)3 complexes. 4. Competition between pyridine- and pyrrole-type substrates toward B(C6F5)3: structure and dynamics of 7-B(C6F5)3-7-azaindole and [7-Azaindolium]+[HOB(C6F5)3]-.

    PubMed

    Focante, Francesca; Camurati, Isabella; Resconi, Luigi; Guidotti, Simona; Beringhelli, Tiziana; D'Alfonso, Giuseppe; Donghi, Daniela; Maggioni, Daniela; Mercandelli, Pierluigi; Sironi, Angelo

    2006-02-20

    Reaction between 7-azaindole and B(C6F5)3 quantitatively yields 7-(C6F5)3B-7-azaindole (4), in which B(C6F5)3 coordinates to the pyridine nitrogen of 7-azaindole, leaving the pyrrole ring unreacted even in the presence of a second equivalent of B(C6F5)3. Reaction of 7-azaindole with H2O-B(C6F5)3 initially produces [7-azaindolium]+[HOB(C6F5)3]- (5) which slowly converts to 4 releasing a H2O molecule. Pyridine removes the borane from the known complexes (C6F5)3B-pyrrole (1) and (C6F5)3B-indole (2), with formation of free pyrrole or indole, giving the more stable adduct (C6F5)3B-pyridine (3). The competition between pyridine and 7-azaindole for the coordination with B(C6F5)3 again yields 3. The molecular structures of compounds 4 and 5 have been determined both in the solid state and in solution and compared to the structures of other (C6F5)3B-N-heterocycle complexes. Two dynamic processes have been found in compound 4. Their activation parameters (DeltaH = 66 (3) kJ/mol, DeltaS = -18 (10) J/mol K and DeltaH = 76 (5) kJ/mol, DeltaS = -5 (18) J/mol K) are comparable with those of other (C6F5)3B-based adducts. The nature of the intramolecular interactions that result in such energetic barriers is discussed. PMID:16471981

  5. Structural insights into human Kif7, a kinesin involved in Hedgehog signalling

    SciTech Connect

    Klejnot, Marta Kozielski, Frank

    2012-02-01

    The human Kif7 motor domain structure provides insights into a kinesin of medical significance. Kif7, a member of the kinesin 4 superfamily, is implicated in a variety of diseases including Joubert, hydrolethalus and acrocallosal syndromes. It is also involved in primary cilium formation and the Hedgehog signalling pathway and may play a role in cancer. Its activity is crucial for embryonic development. Kif7 and Kif27, a closely related kinesin in the same subfamily, are orthologues of the Drosophila melano@@gaster kinesin-like protein Costal-2 (Cos2). In vertebrates, they work together to fulfil the role of the single Cos2 gene in Drosophila. Here, the high-resolution structure of the human Kif7 motor domain is reported and is compared with that of conventional kinesin, the founding member of the kinesin superfamily. These data are a first step towards structural characterization of a kinesin-4 family member and of this interesting molecular motor of medical significance.

  6. Old Maid Flat geothermal exploratory hole No. 7A drilling and completion report

    SciTech Connect

    Not Available

    1981-02-01

    Drilling and testing efforts for a 6000-foot geothermal exploratory hole on the western approaches to Mount Hood, near Portland, Oregon were completed. The intent of the drilling was to encounter a hydrothermal reservoir in a postulated fracture system and confirm the existence of a moderate-temperature (200/sup 0/F) geothermal resource in the Old Main Flat (OMF) vicinity of Mount Hood. The exploratory hole, OMF No. 7A, was completed to a total depth of 6027 feet in 54 days using conventional rotary drilling techniques. The hole was found to be incapable of producing fluids with the desired temperatures. A maximum hole temperature of about 235/sup 0/F was recorded at total depth and a temperature gradient of about 3.3/sup 0/F/100 feet was exhibited over the lower 1000 feet of hole. A variety of technical data, including physical samples such as cores, cuttings, and borehole fluids, plus geophysical well logs were acquired. Data analyses are continuing, with results to be made available through future separate reports.

  7. slc7a6os Gene Plays a Critical Role in Defined Areas of the Developing CNS in Zebrafish

    PubMed Central

    Benini, Anna; Cignarella, Francesca; Calvarini, Laura; Mantovanelli, Silvia; Giacopuzzi, Edoardo; Zizioli, Daniela; Borsani, Giuseppe

    2015-01-01

    The aim of this study is to shed light on the functional role of slc7a6os, a gene highly conserved in vertebrates. The Danio rerio slc7a6os gene encodes a protein of 326 amino acids with 46% identity to human SLC7A6OS and 14% to Saccharomyces cerevisiae polypeptide Iwr1. Yeast Iwr1 specifically binds RNA pol II, interacts with the basal transcription machinery and regulates the transcription of specific genes. In this study we investigated for the first time the biological role of SLC7A6OS in vertebrates. Zebrafish slc7a6os is a maternal gene that is expressed throughout development, with a prevalent localization in the developing central nervous system (CNS). The gene is also expressed, although at different levels, in various tissues of the adult fish. To determine the functional role of slc7a6os during zebrafish development, we knocked-down the gene by injecting a splice-blocking morpholino. At 24 hpf morphants show morphological defects in the CNS, particularly the interface between hindbrain and midbrain is not well-defined. At 28 hpf the morpholino injected embryos present an altered somite morphology and appear partially or completely immotile. At this stage the midbrain, hindbrain and cerebellum are compromised and not well defined compared with control embryos. The observed alterations persist at later developmental stages. Consistently, the expression pattern of two markers specifically expressed in the developing CNS, pax2a and neurod, is significantly altered in morphants. The co-injection of embryos with synthetic slc7a6os mRNA, rescues the morphant phenotype and restores the wild type expression pattern of pax2a and neurod. Our data suggest that slc7a6os might play a critical role in defined areas of the developing CNS in vertebrates, probably by regulating the expression of key genes. PMID:25803583

  8. Final evaluation & test report for the standard waste box (docket 01-53-7A) type A packaging

    SciTech Connect

    KELLY, D L

    2001-10-15

    This report documents the U.S. Department of Transportation Specification 7A Type A compliance test and evaluation results of the Standard Waste Box. Testing and evaluation activities documented herein are on behalf of the U.S. Department of Energy-Headquarters, Office of Safety, Health and Security (EM-5), Germantown, Maryland. Duratek Federal Services, Inc., Northwest Operations performed an evaluation of the changes as documented herein under Docket 01-53-7A.

  9. Involvement of CTR1 and ATP7A in lead (Pb)-induced copper (Cu) accumulation in choroidal epithelial cells.

    PubMed

    Zheng, Gang; Zhang, Jieqiong; Xu, Yan; Shen, Xuefeng; Song, Han; Jing, Jinfei; Luo, Wenjing; Zheng, Wei; Chen, Jingyuan

    2014-02-10

    The blood-cerebrospinal fluid barrier (BCB) plays a key role in maintaining copper (Cu) homeostasis in the brain. Cumulative evidences indicate that lead (Pb) exposure alters cerebral Cu homeostasis, which may underlie the development of neurodegenerative diseases. This study investigated the roles of Cu transporter 1 (CTR1) and ATP7A, two Cu transporters, in Pb-induced Cu accumulation in the choroidal epithelial cells. Pb exposure resulted in increased intracellular (64)Cu retention, accompanying with up-regulated CTR1 level. Knockdown of CTR1 using siRNA before Pb exposure diminished the Pb-induced increase of (64)Cu uptake. The expression level of ATP7A was down-regulated following the Pb exposure. ATP7A siRNA knockdown, or PCMB treatment, inhibited the (64)Cu efflux from the cells, while the following additional incubation with Pb failed to further increase the intracellular (64)Cu retention. Cu exposure, or intracellular Cu accumulation following the tetracycline (Tet)-induced overexpression of CTR1, did not result in significant change in ATP7A expression. Taken together, these data indicate that CTR1 and ATP7A play important roles in Cu transport in choroidal epithelial cells, and the Pb-induced intracellular Cu accumulation appears to be mediated, at least in part, via the alteration of CTR1 and ATP7A expression levels following Pb exposure.

  10. The astrocytic transporter SLC7A10 (Asc-1) mediates glycinergic inhibition of spinal cord motor neurons

    PubMed Central

    Ehmsen, Jeffrey T.; Liu, Yong; Wang, Yue; Paladugu, Nikhil; Johnson, Anna E.; Rothstein, Jeffrey D.; du Lac, Sascha; Mattson, Mark P.; Höke, Ahmet

    2016-01-01

    SLC7A10 (Asc-1) is a sodium-independent amino acid transporter known to facilitate transport of a number of amino acids including glycine, L-serine, L-alanine, and L-cysteine, as well as their D-enantiomers. It has been described as a neuronal transporter with a primary role related to modulation of excitatory glutamatergic neurotransmission. We find that SLC7A10 is substantially enriched in a subset of astrocytes of the caudal brain and spinal cord in a distribution corresponding with high densities of glycinergic inhibitory synapses. Accordingly, we find that spinal cord glycine levels are significantly reduced in Slc7a10-null mice and spontaneous glycinergic postsynaptic currents in motor neurons show substantially diminished amplitudes, demonstrating an essential role for SLC7A10 in glycinergic inhibitory function in the central nervous system. These observations establish the etiology of sustained myoclonus (sudden involuntary muscle movements) and early postnatal lethality characteristic of Slc7a10-null mice, and implicate SLC7A10 as a candidate gene and auto-antibody target in human hyperekplexia and stiff person syndrome, respectively. PMID:27759100

  11. Premature termination codons in the Type VII collagen gene (COL7A1) underlie severe, mutilating recessive dystrophic epidermolysis bullosa

    SciTech Connect

    Christiano, A.M.; Uitto, J. ); Anhalt, G. ); Gibbons, S.; Bauer, E.A. )

    1994-05-01

    Epidermolysis bullosa (EB) is a group of heritable mechano-bullous skin diseases classified into three major categories on the basis of the level of tissue separation within the dermal-epidermal basement membrane zone. The most severe, dystrophic (scarring) forms of EB demonstrate blister formation below the cutaneous basement membrane at the level of the anchoring fibrils. Ultrastructural observations of altered anchoring fibrils and genetic linkage to the gene encoding type VII collagen (COL7A1), the major component of anchoring fibrils, have implicated COL7A1 as the candidate gene in the dystrophic forms of EB. The authors have recently cloned the entire cDNA and gene for human COL7A1, which has been mapped to 3p21. In this study, they describe mutations in four COL7A1 alleles in three patients with severe, mutilating recessive dystrophic EB (Hallopeau-Siemens type, HS-RDEB). Each of these mutations resulted in a premature termination codon (PTC) in the amino-terminal portion of COL7A1. One of the patients was a compound heterozygote for two different mutations. The heterozygous carriers showed an [approximately] 50% reduction in anchoring fibrils, yet were clinically unaffected. Premature termination codons in both alleles of COL7A1 may thus be a major underlying cause of the severe, recessive dystrophic forms of EB. 40 refs., 8 figs.

  12. Endosulfan affects uterine development and functional differentiation by disrupting Wnt7a and β-catenin expression in rats.

    PubMed

    Ingaramo, Paola I; Milesi, María M; Schimpf, Marlise Guerrero; Ramos, Jorge G; Vigezzi, Lucía; Muñoz-de-Toro, Mónica; Luque, Enrique H; Varayoud, Jorgelina

    2016-04-15

    Neonatal exposure to a low dose of endosulfan may disrupt the expression of Wnt7a and β-catenin during uterine development leading to the failure of uterine functional differentiation during implantation. New-born female Wistar rats were treated with vehicle, endosulfan (600 μg/kg/d, E600) or diethylstilbestrol (0.2 μg/kg/d, DES) on postnatal days (PNDs) 1, 3, 5 and 7. Subsequently, uterine histomorphology and the protein expression of Wnt7a and β-catenin were evaluated on PND8, PND21 and gestational day (GD) 5 (pre-implantation period). In the E600 rats, Wnt7a and β-catenin protein expression was increased in the epithelium on PND8, and Wnt7a expression was decreased in the endometrial glands on PND21. On GD5, the number of uterine glands was decreased in the E600-and DES-treated rats. In addition, Wnt7a expression was decreased in all uterine compartments, and β-catenin expression was increased in the luminal and glandular epithelia of the E600-and DES-treated rats. Disruption of Wnt7a and β-catenin uterine expression in the prepubertal and adult females altered the uterine preparation for embryo implantation, which could be associated with the subfertility triggered by endosulfan. PMID:26911934

  13. ADAMTS-7: a metalloproteinase that directly binds to and degrades cartilage oligomeric matrix protein

    PubMed Central

    Liu, Chuan-ju; Kong, Wei; Ilalov, Kiril; Yu, Shuang; Xu, Ke; Prazak, Lisa; Fajardo, Marc; Sehgal, Bantoo; Di Cesare, Paul E.

    2006-01-01

    Degradative fragments of cartilage oligomeric matrix protein (COMP) have been observed in arthritic patients. The physiological enzyme(s) that degrade COMP, however, remain unknown. We performed a yeast two-hybrid screen (Y2H) to search for proteins that associate with COMP to identify an interaction partner that might degrade it. One screen using the epidermal growth factor (EGF) domain of COMP as bait led to the discovery of ADAMTS-7. Rat ADAMTS-7 is composed of 1595 amino acids, and this protein exhibits higher expression in the musculoskeletal tissues. COMP binds directly to ADAMTS-7 in vitro and in native articular cartilage. ADAMTS-7 selectively interacts with the EGF repeat domain but not with the other three functional domains of COMP, whereas the four C-terminal TSP motifs of ADAMTS-7 are required and sufficient for association with COMP. The recombinant catalytic domain and intact ADAMTS-7 are capable of digesting COMP in vitro. The enzymatic activity of ADAMTS-7 requires the presence of Zn2+ and appropriate pH (7.5-9.5), and the concentration of ADAMTS-7 in cartilage and synovium of patients with rheumatoid arthritis is significantly increased as compared to normal cartilage and synovium. ADAMTS-7 is the first metalloproteinase found to bind directly to and degrade COMP.—Liu, C., Kong, W., Ilalov, K., Yu, S., Xu, K., Prazak, L., Fajardo, M., Sehgal, B., Di Cesare, P. E. ADAMTS-7: a metalloproteinase that directly binds to and degrades cartilage oligomeric matrix protein. FASEB J. 20, E129 -E140 (2006) PMID:16585064

  14. Bovine CLEC7A genetic variants and their association with seropositivity in Johne's disease ELISA.

    PubMed

    Pant, S D; Verschoor, C P; Schenkel, F S; You, Q; Kelton, D F; Karrow, N A

    2014-03-10

    Mycobacterium avium ssp. paratuberculosis (MAP) infection in cattle causes significant economic losses to the dairy and beef industries resulting from reduced productivity, premature culling and mortality. Bovine Dectin-1, an important pattern recognition molecule that is able to generate a proinflammatory response by acting alongside Toll like receptor (TLR) signaling, is known to co-operate with TLR2 to specifically activate a macrophage proinflammatory response against mycobacterial infections. Therefore, the goal of this study was to identify single nucleotide polymorphisms (SNPs) in the gene encoding bovine Dectin-1 (CLEC7A) and to assess their association with susceptibility to MAP infection in dairy cattle. Blood and milk samples, collected from commercial dairy operations, were tested for MAP infection using blood and milk ELISAs and a resource population consisting of 197 infected and 242 healthy cattle was constructed. Pooled DNA was used for sequencing and eight single nucleotide polymorphisms (SNPs) were identified. Identified SNPs were genotyped on the resource population using the iPLEX MassARRAY system and statistical analysis was performed using logistic regression fitting the additive and dominance effects of each SNP in the model. Out of a total of eight identified SNPs, five were successfully genotyped, and three out of these five SNPs were found to be in complete linkage. Statistical analysis revealed a strong association between a non-synonymous SNP c.589A>G (p=0.008), and MAP infection status of the resource population inferred by seropositivity in MAP antibody specific ELISAs. This SNP c.589A>G was located in the geneic region that encodes the carbohydrate recognition domain of bovine Dectin-1. Therefore, further investigation of its functional relevance is warranted. PMID:24393710

  15. Synthesis and structural characterization of Al{sub 7}C{sub 3}N{sub 3}-homeotypic aluminum silicon oxycarbonitride, (Al{sub 7-x}Si{sub x})(O{sub y}C{sub z}N{sub 6-y-z}) (x{approx}1.2, y{approx}1.0 and z{approx}3.5)

    SciTech Connect

    Urushihara, Daisuke; Kaga, Motoaki; Asaka, Toru; Nakano, Hiromi; Fukuda, Koichiro

    2011-08-15

    A new aluminum silicon oxycarbonitride, (Al{sub 5.8}Si{sub 1.2})(O{sub 1.0}C{sub 3.5}N{sub 1.5}), has been synthesized and characterized by X-ray powder diffraction (XRPD), transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDX) and electron energy loss spectroscopy (EELS). The title compound is hexagonal with space group P6{sub 3}/mmc and unit-cell dimensions a=0.322508(4) nm, c=3.17193(4) nm and V=0.285717(6) nm{sup 3}. The atom ratios of Al:Si and those of O:C:N were, respectively, determined by EDX and EELS. The initial structural model was successfully derived from the XRPD data by the direct methods and further refined by the Rietveld method. The crystal is most probably composed of four types of domains with nearly the same fraction, each of which is isotypic to Al{sub 7}C{sub 3}N{sub 3} with space group P6{sub 3}mc. The existence of another new oxycarbonitride (Al{sub 6.6}Si{sub 1.4})(O{sub 0.7}C{sub 4.3}N{sub 2.0}), which must be homeotypic to Al{sub 8}C{sub 3}N{sub 4}, has been also demonstrated by XRPD and TEM. - Graphical abstract: A new oxycarbonitride discovered in the Al-Si-O-C-N system, (Al{sub 7-x}Si{sub x})(O{sub y}C{sub z}N{sub 6-y-z}) (x{approx}1.2, y{approx}1.0 and z{approx}3.5). The crystal is composed of four types of domains (I, II, III and IV), and hence the structure is represented by a split-atom model. Individual crystal structures can be regarded as layered structures, which consist of A-type [(Al, Si){sub 4}(O, C, N){sub 4}] unit layers and B-type [(Al, Si)(O, C, N){sub 2}] single layers. Highlights: > (Al{sub 5.8}Si{sub 1.2})(O{sub 1.0}C{sub 3.5}N{sub 1.5}) as a new aluminum silicon oxycarbonitride. > Crystal structure is determined and represented by a split-atom model. > Existence of another new oxycarbonitride (Al{sub 6.6}Si{sub 1.4})(O{sub 0.7}C{sub 4.3}N{sub 2.0}) is demonstrated. > Both new materials are formed by oxidation and nitridation of (Al, Si){sub 6}(O, C){sub 5}.

  16. Wnt7a is a novel inducer of β-catenin-independent tumor-suppressive cellular senescence in lung cancer

    PubMed Central

    Bikkavilli, R K; Avasarala, S; Van Scoyk, M; Arcaroli, J; Brzezinski, C; Zhang, W; Edwards, M G; Rathinam, M K K; Zhou, T; Tauler, J; Borowicz, S; Lussier, Y A; Parr, B A; Cool, C D; Winn, R A

    2015-01-01

    Cellular senescence is an initial barrier for carcinogenesis. However, the signaling mechanisms that trigger cellular senescence are incompletely understood, particularly in vivo. Here we identify Wnt7a as a novel upstream inducer of cellular senescence. In two different mouse strains (C57Bl/6J and FVB/NJ), we show that the loss of Wnt7a is a major contributing factor for increased lung tumorigenesis owing to reduced cellular senescence, and not reduced apoptosis, or autophagy. Wnt7a-null mice under de novo conditions and in both the strains display E-cadherin-to-N-cadherin switch, reduced expression of cellular senescence markers and reduced expression of senescence-associated secretory phenotype, indicating a genetic predisposition of these mice to increased carcinogen-induced lung tumorigenesis. Interestingly, Wnt7a induced an alternate senescence pathway, which was independent of β-catenin, and distinct from that of classical oncogene-induced senescence mediated by the well-known p16INK4a and p19ARF pathways. Mechanistically, Wnt7a induced cellular senescence via inactivation of S-phase kinase-associated protein 2, an important alternate regulator of cellular senescence. Additionally, we identified Iloprost, a prostacyclin analog, which initiates downstream signaling cascades similar to that of Wnt7a, as a novel inducer of cellular senescence, presenting potential future clinical translational strategies. Thus pro-senescence therapies using either Wnt7a or its mimic, Iloprost, might represent a new class of therapeutic treatments for lung cancer. PMID:25728679

  17. Wnt7a is a novel inducer of β-catenin-independent tumor-suppressive cellular senescence in lung cancer.

    PubMed

    Bikkavilli, R K; Avasarala, S; Van Scoyk, M; Arcaroli, J; Brzezinski, C; Zhang, W; Edwards, M G; Rathinam, M K K; Zhou, T; Tauler, J; Borowicz, S; Lussier, Y A; Parr, B A; Cool, C D; Winn, R A

    2015-10-16

    Cellular senescence is an initial barrier for carcinogenesis. However, the signaling mechanisms that trigger cellular senescence are incompletely understood, particularly in vivo. Here we identify Wnt7a as a novel upstream inducer of cellular senescence. In two different mouse strains (C57Bl/6J and FVB/NJ), we show that the loss of Wnt7a is a major contributing factor for increased lung tumorigenesis owing to reduced cellular senescence, and not reduced apoptosis, or autophagy. Wnt7a-null mice under de novo conditions and in both the strains display E-cadherin-to-N-cadherin switch, reduced expression of cellular senescence markers and reduced expression of senescence-associated secretory phenotype, indicating a genetic predisposition of these mice to increased carcinogen-induced lung tumorigenesis. Interestingly, Wnt7a induced an alternate senescence pathway, which was independent of β-catenin, and distinct from that of classical oncogene-induced senescence mediated by the well-known p16(INK4a) and p19(ARF) pathways. Mechanistically, Wnt7a induced cellular senescence via inactivation of S-phase kinase-associated protein 2, an important alternate regulator of cellular senescence. Additionally, we identified Iloprost, a prostacyclin analog, which initiates downstream signaling cascades similar to that of Wnt7a, as a novel inducer of cellular senescence, presenting potential future clinical translational strategies. Thus pro-senescence therapies using either Wnt7a or its mimic, Iloprost, might represent a new class of therapeutic treatments for lung cancer. PMID:25728679

  18. Cyclopalladated Compound 7a Induces Apoptosis- and Autophagy-Like Mechanisms in Paracoccidioides and Is a Candidate for Paracoccidioidomycosis Treatment.

    PubMed

    Arruda, Denise C; Matsuo, Alisson L; Silva, Luiz S; Real, Fernando; Leitão, Natanael P; Pires, Jhon H S; Caires, Antonio Carlos F; Garcia, Daniel M; Cunha, Fernanda F M; Puccia, Rosana; Longo, Larissa V G

    2015-12-01

    Paracoccidioidomycosis (PCM), caused by Paracoccidioides species, is the main cause of death due to systemic mycoses in Brazil and other Latin American countries. Therapeutic options for PCM and other systemic mycoses are limited and time-consuming, and there are high rates of noncompliance, relapses, toxic side effects, and sequelae. Previous work has shown that the cyclopalladated 7a compound is effective in treating several kinds of cancer and parasitic Chagas disease without significant toxicity in animals. Here we show that cyclopalladated 7a inhibited the in vitro growth of Paracoccidioides lutzii Pb01 and P. brasiliensis isolates Pb18 (highly virulent), Pb2, Pb3, and Pb4 (less virulent) in a dose-response manner. Pb18 was the most resistant. Opportunistic Candida albicans and Cryptococcus neoformans were also sensitive. BALB/c mice showed significantly lighter lung fungal burdens when treated twice a day for 20 days with a low cyclopalladated 7a dose of 30 μg/ml/day for 30 days after intratracheal infection with Pb18. Electron microscopy images suggested that apoptosis- and autophagy-like mechanisms are involved in the fungal killing mechanism of cyclopalladated 7a. Pb18 yeast cells incubated with the 7a compound showed remarkable chromatin condensation, DNA degradation, superoxide anion production, and increased metacaspase activity suggestive of apoptosis. Autophagy-related killing mechanisms were suggested by increased autophagic vacuole numbers and acidification, as indicated by an increase in LysoTracker and monodansylcadaverine (MDC) staining in cyclopalladated 7a-treated Pb18 yeast cells. Considering that cyclopalladated 7a is highly tolerated in vivo and affects yeast fungal growth through general apoptosis- and autophagy-like mechanisms, it is a novel promising drug for the treatment of PCM and other mycoses. PMID:26349827

  19. Effects of ATP7A overexpression in mice on copper transport and metabolism in lactation and gestation.

    PubMed

    Wadwa, Jarrod; Chu, Yu-Hsiang; Nguyen, Nhu; Henson, Thomas; Figueroa, Alyssa; Llanos, Roxana; Ackland, Margaret Leigh; Michalczyk, Agnes; Fullriede, Hendrik; Brennan, Grant; Mercer, Julian F B; Linder, Maria C

    2014-01-01

    Placentae and mammary epithelial cells are unusual in robustly expressing two copper "pumps", ATP7A and B, raising the question of their individual roles in these tissues in pregnancy and lactation. Confocal microscopic evidence locates ATP7A to the fetal side of syncytiotrophoblasts, suggesting a role in pumping Cu towards the fetus; and to the basolateral (blood) side of lactating mammary epithelial cells, suggesting a role in recycling Cu to the blood. We tested these concepts in wild-type C57BL6 mice and their transgenic counterparts that expressed hATP7A at levels 10-20× those of endogenous mAtp7a. In lactation, overexpression of ATP7A reduced the Cu concentrations of the mammary gland and milk ~50%. Rates of transfer of tracer (64)Cu to the suckling pups were similarly reduced over 30-48 h, as was the total Cu in 10-day -old pups. During the early and middle periods of gestation, the transgenic litters had higher Cu concentrations than the wild-type, placental Cu showing the reverse trend; but this difference was lost by the first postnatal day. The transgenic mice expressed ATP7A in some hepatocytes, so we investigated the possibility that metalation of ceruloplasmin (Cp) might be enhanced. Rates of (64)Cu incorporation into Cp, oxidase activity, and ratios of holo to apoceruloplasmin were unchanged. We conclude that in the lactating mammary gland, the role of ATP7A is to return Cu to the blood, while in the placenta it mediates Cu delivery to the fetus and is the rate-limiting step for fetal Cu nutrition during most of gestation in mice. PMID:24744874

  20. Effects of ATP7A overexpression in mice on copper transport and metabolism in lactation and gestation

    PubMed Central

    Wadwa, Jarrod; Chu, Yu‐Hsiang; Nguyen, Nhu; Henson, Thomas; Figueroa, Alyssa; Llanos, Roxana; Ackland, Margaret Leigh; Michalczyk, Agnes; Fullriede, Hendrik; Brennan, Grant; Mercer, Julian F. B.; Linder, Maria C.

    2014-01-01

    Abstract Placentae and mammary epithelial cells are unusual in robustly expressing two copper “pumps”, ATP7A and B, raising the question of their individual roles in these tissues in pregnancy and lactation. Confocal microscopic evidence locates ATP7A to the fetal side of syncytiotrophoblasts, suggesting a role in pumping Cu towards the fetus; and to the basolateral (blood) side of lactating mammary epithelial cells, suggesting a role in recycling Cu to the blood. We tested these concepts in wild‐type C57BL6 mice and their transgenic counterparts that expressed hATP7A at levels 10–20× those of endogenous mAtp7a. In lactation, overexpression of ATP7A reduced the Cu concentrations of the mammary gland and milk ~50%. Rates of transfer of tracer 64Cu to the suckling pups were similarly reduced over 30–48 h, as was the total Cu in 10‐day ‐old pups. During the early and middle periods of gestation, the transgenic litters had higher Cu concentrations than the wild‐type, placental Cu showing the reverse trend; but this difference was lost by the first postnatal day. The transgenic mice expressed ATP7A in some hepatocytes, so we investigated the possibility that metalation of ceruloplasmin (Cp) might be enhanced. Rates of 64Cu incorporation into Cp, oxidase activity, and ratios of holo to apoceruloplasmin were unchanged. We conclude that in the lactating mammary gland, the role of ATP7A is to return Cu to the blood, while in the placenta it mediates Cu delivery to the fetus and is the rate‐limiting step for fetal Cu nutrition during most of gestation in mice. PMID:24744874

  1. Dynamic Regulation of Wnt7a Expression in the Primate Endometrium: Implications for Postmenstrual Regeneration and Secretory Transformation

    PubMed Central

    Fan, Xiujun; Krieg, Sacha; Hwang, Jong Yun; Dhal, Sabita; Kuo, Calvin J.; Lasley, Bill L.; Brenner, Robert M.

    2012-01-01

    Despite the vital physiological role of endometrial regeneration during the menstrual cycle and the various pathological implications of abnormal growth of endometrial epithelial cells, the local factors and regulatory mechanisms involved in endometrial regeneration and growth have not been well characterized. Here, we examine the pattern, hormone dependence, and potential functions of Wnt7a (wingless-type MMTV integration site family member 7a), which is known to play a critical role in the formation of the mouse endometrial epithelium during embryonic development, in both human and artificially cycling rhesus macaque endometrium, and using a potent Wnt-antagonist in a mouse model of endometrial regeneration. Wnt7a transcript levels were examined using quantitative real-time PCR and in situ hybridization, and immunohistochemistry was performed to detect Ki-67 and 3,5-bromodeoxyuridine. Stringent, fully conditional Wnt inhibition was achieved by adenoviral expression of Dickkopf-1 during artificial endometrial regeneration in mice. In macaques, Wnt7a expression was confined to the newly formed luminal epithelium (LE) and upper glands during the postmenstrual repair phase. The signal increased in the LE during the proliferative phase but decreased in the upper glands and was undetectable in the glands by the late proliferative phase. Interestingly, Wnt7a was completely suppressed in the LE and remained undetectable in other cell types after 7 d of progesterone treatment. The pattern of Wnt7a expression in the human endometrium was similar to that in macaques. Blockade of Wnt signaling during endometrial regeneration in mice resulted in a dramatic delay in reepithelialization and degeneration of glands and LE. These results strongly suggest, for the first time, a role for Wnt7a in postmenstrual regeneration and proliferation of endometrial glands and LE in primates, and its dramatic suppression by progesterone is likely essential for secretory transformation of the

  2. UV testing of INTELSAT-7, 7A, and 8 solar cells

    NASA Technical Reports Server (NTRS)

    Meulenberg, A.

    1994-01-01

    A 4000 hour experiment, conducted in late 1992 through mid 1993, confirmed earlier results on the ultraviolet damage effects in covered solar cells of various types being used, or proposed for use, in INTELSAT programs. Two different UV test systems were used to identify systematic errors and to study the effects of UV source-bulb age on degradation rate. After correction for contamination and UV source-bulb aging, the extrapolated degradation rates for irradiated and unirradiated INTELSAT-5, -6 single AR(SAR) coated cells and INTELSAT-7, -7A, -8 double layer AR(DAR) coated cells in both the 1993 tests confirm the following hypotheses resulting from the 1992 experiment. (a) Irradiated cells display significantly more UV degradation than do the unirradiated cells for tests exceeding 2000 hours. The new data indicates that degradation effects from electron irradiation are proportional to t(exp 2) (the square of the UV hours), at least for times less than or equal to 3000 hours. (b) This difference does not depend upon entire reflective coating, cell resistivity, or manufacturer within the sensitivity and reproducibility of the experiment. (c) There is a clear difference in degradation rate between single AR coated cells (TiO(x)) and double layer AR coated cells (SiO(x) and Al2O3?). At 100,000 hours (11.4 years) the DAR coated cells display more degradation than do the SAR coated cells, even though at 1,000 hours the DAR cells display less degradation. (d) UV degradation rates, to modern covered silicon solar cells, at the beginning of bulb life drop from approximately 2 times the average rate to near zero after 2000 hours (average end-of-life for the xenon short-arc lamps used in the tests). The effects of 1 MeV electron irradiation (10(exp 15) e(-)/sq cm) prior to UV exposure are clearly indicated in the plot of percent change in cell open circuit voltage (Voc) versus percent change in short circuit current (Isc) during the UV test and post-test cleanup of the cells

  3. Innexin7a forms junctions that stabilize the basal membrane during cellularization of the blastoderm in Tribolium castaneum.

    PubMed

    van der Zee, Maurijn; Benton, Matthew A; Vazquez-Faci, Tania; Lamers, Gerda E M; Jacobs, Chris G C; Rabouille, Catherine

    2015-06-15

    In insects, the fertilized egg undergoes a series of rapid nuclear divisions before the syncytial blastoderm starts to cellularize. Cellularization has been extensively studied in Drosophila melanogaster, but its thick columnar blastoderm is unusual among insects. We therefore set out to describe cellularization in the beetle Tribolium castaneum, the embryos of which exhibit a thin blastoderm of cuboidal cells, like most insects. Using immunohistochemistry, live imaging and transmission electron microscopy, we describe several striking differences to cellularization in Drosophila, including the formation of junctions between the forming basal membrane and the yolk plasmalemma. To identify the nature of this novel junction, we used the parental RNAi technique for a small-scale screen of junction proteins. We find that maternal knockdown of Tribolium innexin7a (Tc-inx7a), an ortholog of the Drosophila gap junction gene Innexin 7, leads to failure of cellularization. In Inx7a-depleted eggs, the invaginated plasma membrane retracts when basal cell closure normally begins. Furthermore, transiently expressed tagged Inx7a localizes to the nascent basal membrane of the forming cells in wild-type eggs. We propose that Inx7a forms the newly identified junctions that stabilize the forming basal membrane and enable basal cell closure. We put forward Tribolium as a model for studying a more ancestral mode of cellularization in insects.

  4. [Expression levels of Slc7a11 in the skin of Kazakh sheep with different coat colors].

    PubMed

    Li, Hong-Tao; He, Xin; Zhou, Zhi-Yong; Zhao, Song-Hua; Zhang, Wen-Xiang; Liu, Gang; Zhao, Zong-Sheng; Jia, Bin

    2012-10-01

    Slc7a11 belongs to solute transporter gene family, encoding cystine/glutamate transporter xCT. It regulates switching between eumelanin and pheomelanin synthesis. In the present study, Real-time PCR was used to detect the mRNA expression levels of Slc7a11 in the skin of Kazakh lambs with different coat colors (black, brown and white), and then the prokaryotic expression plasmid PET-32a-sxCT was constructed to induce the expression of fusion protein. The target pro-tein was purified by Ni-NTA affinity chromatographic separation, and then was used to immunize rabbit in order to produce rabbit anti-sxCT polyclonal antibody. Finally, the expression levels of sxCT were detected in the skin of Kazakh lambs with different hair colors by Western blotting analysis. Results showed that the mRNA expression levels of Slc7a11 differed significantly in the skin of Kazakh lambs with different coat colors, with the highest level in brown coat color, followed by the black, and then the white. The sxCT protein was also detected in the skin of different coat colors by polyclonal antibody, with the highest level in brown coat color, followed by the black, and then the white. It is, therefore, concluded that slc7a11 gene might be associated with the phenotype of coat color in Kazakh sheep.

  5. [Expression levels of Slc7a11 in the skin of Kazakh sheep with different coat colors].

    PubMed

    Li, Hong-Tao; He, Xin; Zhou, Zhi-Yong; Zhao, Song-Hua; Zhang, Wen-Xiang; Liu, Gang; Zhao, Zong-Sheng; Jia, Bin

    2012-10-01

    Slc7a11 belongs to solute transporter gene family, encoding cystine/glutamate transporter xCT. It regulates switching between eumelanin and pheomelanin synthesis. In the present study, Real-time PCR was used to detect the mRNA expression levels of Slc7a11 in the skin of Kazakh lambs with different coat colors (black, brown and white), and then the prokaryotic expression plasmid PET-32a-sxCT was constructed to induce the expression of fusion protein. The target pro-tein was purified by Ni-NTA affinity chromatographic separation, and then was used to immunize rabbit in order to produce rabbit anti-sxCT polyclonal antibody. Finally, the expression levels of sxCT were detected in the skin of Kazakh lambs with different hair colors by Western blotting analysis. Results showed that the mRNA expression levels of Slc7a11 differed significantly in the skin of Kazakh lambs with different coat colors, with the highest level in brown coat color, followed by the black, and then the white. The sxCT protein was also detected in the skin of different coat colors by polyclonal antibody, with the highest level in brown coat color, followed by the black, and then the white. It is, therefore, concluded that slc7a11 gene might be associated with the phenotype of coat color in Kazakh sheep. PMID:23099788

  6. 13 CFR 120.431 - Which Lenders may sell, sell participations in, or pledge 7(a) loans?

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 13 Business Credit and Assistance 1 2010-01-01 2010-01-01 false Which Lenders may sell, sell participations in, or pledge 7(a) loans? 120.431 Section 120.431 Business Credit and Assistance SMALL BUSINESS ADMINISTRATION BUSINESS LOANS Lenders Other Conveyances § 120.431 Which Lenders may sell, sell participations...

  7. BMP-2-enhanced chondrogenesis involves p38 MAPK-mediated down-regulation of Wnt-7a pathway.

    PubMed

    Jin, Eun-Jung; Lee, Sun-Young; Choi, Young-Ae; Jung, Jae-Chang; Bang, Ok-Sun; Kang, Shin-Sung

    2006-12-31

    The bone morphogenetic protein (BMP) family has been implicated in control of cartilage development. Here, we demonstrate that BMP-2 promotes chondrogenesis by activating p38 mitogen-activated protein kinase (MAPK), which in turn downregulates Wnt-7a/b-catenin signaling responsible for proteasomal degradation of Sox9. Exposure of mesenchymal cells to BMP-2 resulted in upregulation of Sox9 protein and a concomitant decrease in the level of b-catenin protein and Wnt-7a signaling. In agreement with this, the interaction of Sox9 with b-catenin was inhibited in the presence of BMP-2. Inhibition of the p38 MAPK pathway using a dominant negative mutant led to sustained Wnt-7a signaling and decreased Sox9 expression, with consequent inhibition of precartilage condensation and chondrogenic differentiation. Moreover, overexpression of b-catenin caused degradation of Sox9 via the ubiquitin/26S proteasome pathway. Our results collectively indicate that the increase in Sox9 protein resulting from downregulation of b-catenin/Wnt-7a signaling is mediated by p38 MAPK during BMP-2 induced chondrogenesis in chick wing bud mesenchymal cells. PMID:17202865

  8. CYP7A1 Gene Polymorphism Located in the 5′ Upstream Region Modifies the Risk of Coronary Artery Disease

    PubMed Central

    Iwanicki, Tomasz; Balcerzyk, Anna; Niemiec, Pawel; Nowak, Tomasz; Ochalska-Tyka, Anna; Krauze, Jolanta; Kosiorz-Gorczynska, Sylwia; Grzeszczak, Wladyslaw; Zak, Iwona

    2015-01-01

    Background. 7-Alpha cholesterol hydroxylase (CYP7A1), the first enzyme of classic conversion pathway leading from cholesterol to bile acids synthesis, is encoded by CYP7A1 gene. Its single nucleotide polymorphisms (SNPs) influence serum lipid levels and may be related to impaired lipid profile leading to coronary artery disease (CAD). The aim of the present study was to analyze the possible association between the rs7833904 CYP7A1 polymorphism and premature CAD. Material and Methods. Serum lipid levels and rs7833904 SNP were determined in 419 subjects: 200 patients with premature CAD and 219 age and sex matched controls. Results. The A allele carrier state was associated with CAD (OR = 1.76, 95% CI; 1.14–2.71, P = 0.014). The effect was even stronger in the male subgroups (OR = 2.16, 95% CI; 1.28–3.65, P = 0.003). There was no effect in the females. Risk factors of CAD and clinical phenotype of atherosclerosis were not associated with genotype variants of the rs7833904 SNP. Lipid profiles also did not differ significantly between individual genotypes. Conclusion. The CYP7A1 rs7833904 polymorphism may modify the risk of CAD. This effect is especially strong in male subjects. The studied polymorphism does not significantly influence serum lipid levels, in the present study. PMID:25944972

  9. Engineering of a glycosidase Family 7 cellobiohydrolase to more alkaline pH optimum: the pH behaviour of Trichoderma reesei Cel7A and its E223S/ A224H/L225V/T226A/D262G mutant.

    PubMed Central

    Becker, D; Braet, C; Brumer , H; Claeyssens, M; Divne, C; Fagerström, B R; Harris, M; Jones, T A; Kleywegt, G J; Koivula, A; Mahdi, S; Piens, K; Sinnott, M L; Ståhlberg, J; Teeri, T T; Underwood, M; Wohlfahrt, G

    2001-01-01

    The crystal structures of Family 7 glycohydrolases suggest that a histidine residue near the acid/base catalyst could account for the higher pH optimum of the Humicola insolens endoglucanase Cel7B, than the corresponding Trichoderma reesei enzymes. Modelling studies indicated that introduction of histidine at the homologous position in T. reesei Cel7A (Ala(224)) required additional changes to accommodate the bulkier histidine side chain. X-ray crystallography of the catalytic domain of the E223S/A224H/L225V/T226A/D262G mutant reveals that major differences from the wild-type are confined to the mutations themselves. The introduced histidine residue is in plane with its counterpart in H. insolens Cel7B, but is 1.0 A (=0.1 nm) closer to the acid/base Glu(217) residue, with a 3.1 A contact between N(epsilon2) and O(epsilon1). The pH variation of k(cat)/K(m) for 3,4-dinitrophenyl lactoside hydrolysis was accurately bell-shaped for both wild-type and mutant, with pK(1) shifting from 2.22+/-0.03 in the wild-type to 3.19+/-0.03 in the mutant, and pK(2) shifting from 5.99+/-0.02 to 6.78+/-0.02. With this poor substrate, the ionizations probably represent those of the free enzyme. The relative k(cat) for 2-chloro-4-nitrophenyl lactoside showed similar behaviour. The shift in the mutant pH optimum was associated with lower k(cat)/K(m) values for both lactosides and cellobiosides, and a marginally lower stability. However, k(cat) values for cellobiosides are higher for the mutant. This we attribute to reduced non-productive binding in the +1 and +2 subsites; inhibition by cellobiose is certainly relieved in the mutant. The weaker binding of cellobiose is due to the loss of two water-mediated hydrogen bonds. PMID:11336632

  10. A novel technique for measurement of thermal rate constants and temperature dependences of dissociative recombination: CO2(+), CF3(+), N2O(+), C7H8(+), C7H7(+), C6H6(+), C6H5(+), C5H6(+), C4H4(+), and C3H3(+).

    PubMed

    Fournier, Joseph A; Shuman, Nicholas S; Melko, Joshua J; Ard, Shaun G; Viggiano, Albert A

    2013-04-21

    A novel technique using a flowing afterglow-Langmuir probe apparatus for measurement of temperature dependences of rate constants for dissociative recombination (DR) is presented. Low (~10(11) cm(-3)) concentrations of a neutral precursor are added to a noble gas∕electron afterglow plasma thermalized at 300-500 K. Charge exchange yields one or many cation species, each of which may undergo DR. Relative ion concentrations are monitored at a fixed reaction time while the initial plasma density is varied between 10(9) and 10(10) cm(-3). Modeling of the decrease in concentration of each cation relative to the non-recombining noble gas cation yields the rate constant for DR. The technique is applied to several species (O2(+), CO2(+), CF3(+), N2O(+)) with previously determined 300 K values, showing excellent agreement. The measurements of those species are extended to 500 K, with good agreement to literature values where they exist. Measurements are also made for a range of CnHm(+) (C7H7(+), C7H8(+), C5H6(+), C4H4(+), C6H5(+), C3H3(+), and C6H6(+)) derived from benzene and toluene neutral precursors. CnHm(+) DR rate constants vary from 8-12 × 10(-7) cm(3) s(-1) at 300 K with temperature dependences of approximately T(-0.7). Where prior measurements exist these results are in agreement, with the exception of C3H3(+) where the present results disagree with a previously reported flat temperature dependence. PMID:23614415

  11. Molecular Docking and In Silico ADMET Study Reveals Acylguanidine 7a as a Potential Inhibitor of β-Secretase

    PubMed Central

    Nisha, Chaluveelaveedu Murleedharan; Kumar, Ashwini; Nair, Prateek; Gupta, Nityasha; Silakari, Chitrangda; Tripathi, Timir; Kumar, Awanish

    2016-01-01

    Amyloidogenic pathway in Alzheimer's disease (AD) involves breakdown of APP by β-secretase followed by γ-secretase and results in formation of amyloid beta plaque. β-secretase has been a promising target for developing novel anti-Alzheimer drugs. To test different molecules for this purpose, test ligands like acylguanidine 7a, rosiglitazone, pioglitazone, and tartaric acid were docked against our target protein β-secretase enzyme retrieved from Protein Data Bank, considering MK-8931 (phase III trial, Merck) as the positive control. Docking revealed that, with respect to their free binding energy, acylguanidine 7a has the lowest binding energy followed by MK-8931 and pioglitazone and binds significantly to β-secretase. In silico ADMET predictions revealed that except tartaric acid all other compounds had minimal toxic effects and had good absorption as well as solubility characteristics. These compounds may serve as potential lead compound for developing new anti-Alzheimer drug. PMID:27190510