The upstream enhancer elements of the G6PC promoter are critical for optimal G6PC expression in murine glycogen storage disease type Ia.

Glycogen storage disease type-Ia (GSD-Ia) patients deficient in glucose-6-phosphatase-α (G6Pase-α or G6PC) manifest impaired glucose homeostasis characterized by fasting hypoglycemia, growth retardation, hepatomegaly, nephromegaly, hyperlipidemia, hyperuricemia, and lactic acidemia. Two efficacious recombinant adeno-associated virus pseudotype 2/8 (rAAV8) vectors expressing human G6Pase-α have been independently developed. One is a single-stranded vector containing a 2864-bp of the G6PC promoter/enhancer (rAAV8-GPE) and the other is a double-stranded vector containing a shorter 382-bp minimal G6PC promoter/enhancer (rAAV8-miGPE). To identify the best construct, a direct comparison of the rAAV8-GPE and the rAAV8-miGPE vectors was initiated to determine the best vector to take forward into clinical trials. We show that the rAAV8-GPE vector directed significantly higher levels of hepatic G6Pase-α expression, achieved greater reduction in hepatic glycogen accumulation, and led to a better toleration of fasting in GSD-Ia mice than the rAAV8-miGPE vector. Our results indicated that additional control elements in the rAAV8-GPE vector outweigh the gains from the double-stranded rAAV8-miGPE transduction efficiency, and that the rAAV8-GPE vector is the current choice for clinical translation in human GSD-Ia.

Nanostructured hydroxyapatite surfaces-mediated adsorption alters recognition of BMP receptor IA and bioactivity of bone morphogenetic protein-2

Highly efficient loading of bone morphogenetic protein-2 (BMP-2) onto carriers with desirable performance is still a major challenge in the field of bone regeneration. Till now, the nanoscaled surface-induced changes of the structure and bioactivity of BMP-2 remains poorly understood. Here, the effect of nanoscaled surface on the adsorption and bioactivity of BMP-2 was investigated with a series of hydroxyapatite surfaces (HAPs): HAP crystal-coated surface (HAP), HAP crystal-coated polished surface (HAP-Pol), and sintered HAP crystal-coated surface (HAP-Sin). The adsorption dynamics of recombinant human BMP-2 (rhBMP-2) and the accessibility of the binding epitopes of adsorbed rhBMP-2 for BMP receptors (BMPRs) were examined by a quartz crystal microbalance with dissipation. Moreover, the bioactivity of adsorbed rhBMP-2 and the BMP-induced Smad signaling were investigated with C2C12 model cells. A noticeably high mass-uptake of rhBMP-2 and enhanced recognition of BMPR-IA to adsorbed rhBMP-2 were found on the HAP-Pol surface. For the rhBMP-2-adsorbed HAPs, both ALP activity and Smad signaling increased in the order of HAP-Sin < HAP < HAP-Pol. Furthermore, hybrid molecular dynamics and steered molecular dynamics simulations validated that BMP-2 tightly anchored on the HAP-Pol surface with a relative loosened conformation, but the HAP-Sin surface induced a compact conformation of BMP-2. In conclusion, the nanostructured HAPs can modulate the way of adsorption of rhBMP-2, and thus the recognition of BMPR-IA and the bioactivity of rhBMP-2. These findings can provide insightful suggestions for the future design and fabrication of rhBMP-2-based scaffolds/implants.

Dark-energy constraints and correlations with systematics from CFHTLS weak lensing, SNLS supernovae Ia and WMAP5

Aims We combine measurements of weak gravitational lensing from the CFHTLS-Wide survey, supernovae Ia from CFHT SNLS and CMB anisotropies from WMAP5 to obtain joint constraints on cosmological parameters, in particular, the dark-energy equation-of-state parameter w. We assess the influence of systematics in the data on the results and look for possible correlations with cosmological parameters. Methods We implemented an MCMC algorithm to sample the parameter space of a flat CDM model with a dark-energy component of constant w. Systematics in the data are parametrised and included in the analysis. We determine the influence of photometric calibration of SNIa data on cosmological results by calculating the response of the distance modulus to photometric zero-point variations. The weak lensing data set is tested for anomalous field-to-field variations and a systematic shape measurement bias for high-redshift galaxies. Results Ignoring photometric uncertainties for SNLS biases cosmological parameters by at most 20% of the statistical errors, using supernovae alone; the parameter uncertainties are underestimated by 10%. The weak-lensing field-to-field variance between 1 deg2-MegaCam pointings is 5-15% higher than predicted from N-body simulations. We find no bias in the lensing signal at high redshift, within the framework of a simple model, and marginalising over cosmological parameters. Assuming a systematic underestimation of the lensing signal, the normalisation increases by up to 8%. Combining all three probes we obtain -0.10 < 1 + w < 0.06 at 68% confidence ( -0.18 < 1 + w < 0.12 at 95%), including systematic errors. Our results are therefore consistent with the cosmological constant . Systematics in the data increase the error bars by up to 35%; the best-fit values change by less than 0.15.

Vykopirovka s proektplana Kolaiskoĭ dachi razverstani︠ia︡ Dzhankoĭskoogo raǐona, na uchastky zemli otvedennykh za Nº Nº 106, 109, 66 i 66A

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Vykopirovka so sbornoǐ karty Feodosiǐskoǐ dachi razverstani︠ia︡ Feodosiǐskogo raǐona s ukazaniem svobodnykh fondov na predmet peredachi ikh Komzety

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Vykopirovka s plana Seĭtlerskoĭ dachi razverstani︠ia︡ Kr.A.S.SD.R., na uchastok zemli otvedennyĭ KOMZET'u (Beǐs-Lekhem), pri provedenii rabot po zemreforme v 1930 g.

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Vykopirovka s plana Seĭtlerskoĭ dachi razverstani︠ia︡, Kr.A.S.SD.R., na uchastok zemli otvedennyĭ KOMZET'u (s -Kh. ART. "Rabotnik" - Aranda), pri provedenii rabot po zemreforme v 1930 g.

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Chertezh zemlepol'zovahi︠ia︡ Komzet'a pri der. Kheǐrus, Seǐtlerskogo raǐona, Kr.A.S.SD.R., po proektu zemleustroǐstva 1930 g. po Ichkinskoǐ dache razverstani︠ia︡, v sos.tave Belokoshskogo sevooborotnogo uchastka

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Vykopirovka s plana Seĭtlerskoǐ dachi razverstani︠ia︡ Kr.A.S.SD.R., na uchastok zemli otvedennyĭ KOMZET'u (s.-kh. art. "Vozrazhdenie" - Krasn. Burlak), pri provedenii rabot po zemreforme v 1930 g.

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Vykopirovka s plana Seĭtlerskoǐ dachi razverstani︠ia︡ Kr.A.S.SD.R., na uchastok zemli otvedennyĭ KOMZET'u, pri provedenii rabot po zemreforme v 1930 g.

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Map of water table depths of the Semi-Kolodez︠ia︡nskiǐ fond

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Vykopirovka na uchastok "Kon-Kinchakskiǐ" fond otvedennyǐ v pol'zovanie Komzeta iz Kol-Alchinskoǐ dachi razverstani︠ia︡

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Vykopirovka s plana Semi-Kolodez︠ia︡nskoĭ dachi Kerchenskogo raĭona Oĭsulskogo proizvod. uchastka

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