A common polymorphism in the LDL receptor gene has multiple effects on LDL receptor function.

A common synonymous single nucleotide polymorphism in exon 12 of the low-density lipoprotein receptor (LDLR) gene, rs688, has been associated with increased plasma total and LDL cholesterol in several populations. Using immortalized lymphoblastoid cell lines from a healthy study population, we confirmed an earlier report that the minor allele of rs688 is associated with increased exon 12 alternative splicing (P < 0.05) and showed that this triggered nonsense-mediated decay (NMD) of the alternatively spliced LDLR mRNA. However, since synonymous single nucleotide polymorphisms may influence structure and function of the encoded proteins by co-translational effects, we sought to test whether rs688 was also functional in the full-length mRNA. In HepG2 cells expressing LDLR cDNA constructs engineered to contain the major or minor allele of rs688, the latter was associated with a smaller amount of LDLR protein at the cell surface (-21.8 ± 0.6%, P = 0.012), a higher amount in the lysosome fraction (+25.7 ± 0.3%, P = 0.037) and reduced uptake of fluorescently labeled LDL (-24.3 ± 0.7%, P < 0.01). Moreover, in the presence of exogenous proprotein convertase subtilisin/kexin type 9 (PCSK9), a protein that reduces cellular LDL uptake by promoting lysosomal degradation of LDLR, the minor allele resulted in reduced capacity of a PCSK9 monoclonal antibody to increase LDL uptake. These findings are consistent with the hypothesis that rs688, which is located in the β-propeller region of LDLR, has effects on LDLR activity beyond its role in alternative splicing due to impairment of LDLR endosomal recycling and/or PCSK9 binding, processes in which the β-propeller is critically involved

A common polymorphism in the LDL receptor gene has multiple effects on LDL receptor function.

A common synonymous single nucleotide polymorphism in exon 12 of the low-density lipoprotein receptor (LDLR) gene, rs688, has been associated with increased plasma total and LDL cholesterol in several populations. Using immortalized lymphoblastoid cell lines from a healthy study population, we confirmed an earlier report that the minor allele of rs688 is associated with increased exon 12 alternative splicing (P < 0.05) and showed that this triggered nonsense-mediated decay (NMD) of the alternatively spliced LDLR mRNA. However, since synonymous single nucleotide polymorphisms may influence structure and function of the encoded proteins by co-translational effects, we sought to test whether rs688 was also functional in the full-length mRNA. In HepG2 cells expressing LDLR cDNA constructs engineered to contain the major or minor allele of rs688, the latter was associated with a smaller amount of LDLR protein at the cell surface (-21.8 ± 0.6%, P = 0.012), a higher amount in the lysosome fraction (+25.7 ± 0.3%, P = 0.037) and reduced uptake of fluorescently labeled LDL (-24.3 ± 0.7%, P < 0.01). Moreover, in the presence of exogenous proprotein convertase subtilisin/kexin type 9 (PCSK9), a protein that reduces cellular LDL uptake by promoting lysosomal degradation of LDLR, the minor allele resulted in reduced capacity of a PCSK9 monoclonal antibody to increase LDL uptake. These findings are consistent with the hypothesis that rs688, which is located in the β-propeller region of LDLR, has effects on LDLR activity beyond its role in alternative splicing due to impairment of LDLR endosomal recycling and/or PCSK9 binding, processes in which the β-propeller is critically involved

Biomechanical Analysis of an S1 Pedicle Screw Salvage Technique via a Superior Articulating Process Entry Point

Study designBiomechanical, cadaveric study.ObjectiveTo compare the fixation strength of a novel S1 pedicle screw insertion technique in a revision setting to a standard S1 pedicle screw and an L5 pedicle screw.Summary of background dataFusions to the sacrum remain a difficult clinical challenge. Very few salvage techniques exist when a nonunion occurs.MethodsThe biomechanical integrity of three screw fixations, L5 pedicle screws, a standard S1 pedicle screw, and an S1 pedicle screw placed via a superior articulating process entry point (SAP S1), was characterized by performing pullout tests using cadaveric specimens including L5 and sacrum.ResultsSAP S1 constructs (735.5 ± 110.1 N/mm) were significantly stiffer than standard S1 (P = 0.005) and L5 (P = 0.02) constructs. There was no statistically significant difference between the L5 constructs and the standard S1 constructs for linear stiffness. There was no statistical difference between the three fixations for yield load, displacement at yield load, and energy absorbed to yield load.The ultimate pullout force for the SAP S1 was statistically higher than the standard S1 (1213.7 ± 579.6 vs. 478.6 ± 452.9 N; P = 0.004). Displacement at ultimate load was significantly greater for L5 screw fixation (3.3 ± 1.1 mm) compared to the other two constructs. Both the L5 (2277.4 ± 1873.3 N-mm) and SAP S1 (2628.2 ± 2054.4 N-mm) constructs had significantly greater energy absorbed to ultimate load than the standard S1 construct (811.7 ± 937.6 N-mm), but there was no statistical difference between the L5 and SAP S1 constructs.ConclusionS1 pedicle screw fixation via an SAP entry point provides biomechanical advantages compared to screws placed via the standard S1 or L5 entry point and may be a viable option for revision of a failed L5-S1 fusion with a compromised standard S1 entry point.Level of evidenceN/A

A common polymorphism in the LDL receptor gene has multiple effects on LDL receptor function

A common synonymous single nucleotide polymorphism in exon 12 of the low-density lipoprotein receptor (LDLR) gene, rs688, has been associated with increased plasma total and LDL cholesterol in several populations. Using immortalized lymphoblastoid cell lines from a healthy study population, we confirmed an earlier report that the minor allele of rs688 is associated with increased exon 12 alternative splicing (P < 0.05) and showed that this triggered nonsense-mediated decay (NMD) of the alternatively spliced LDLR mRNA. However, since synonymous single nucleotide polymorphisms may influence structure and function of the encoded proteins by co-translational effects, we sought to test whether rs688 was also functional in the full-length mRNA. In HepG2 cells expressing LDLR cDNA constructs engineered to contain the major or minor allele of rs688, the latter was associated with a smaller amount of LDLR protein at the cell surface (−21.8 ± 0.6%, P = 0.012), a higher amount in the lysosome fraction (+25.7 ± 0.3%, P = 0.037) and reduced uptake of fluorescently labeled LDL (−24.3 ± 0.7%, P < 0.01). Moreover, in the presence of exogenous proprotein convertase subtilisin/kexin type 9 (PCSK9), a protein that reduces cellular LDL uptake by promoting lysosomal degradation of LDLR, the minor allele resulted in reduced capacity of a PCSK9 monoclonal antibody to increase LDL uptake. These findings are consistent with the hypothesis that rs688, which is located in the β-propeller region of LDLR, has effects on LDLR activity beyond its role in alternative splicing due to impairment of LDLR endosomal recycling and/or PCSK9 binding, processes in which the β-propeller is critically involved

Relation between Photoactive Layer Thickness, 3D Morphology, and Device Performance in P3HT/PCBM Bulk-Heterojunction Solar Cells

To get an efficient organic solar cell, as much light as possible should be absorbed by the photoactive layer; as a consequence, thick layers should be preferable. However, it is often observed that much thinner photoactive layers result in more efficient devices than the corresponding thicker layers absorbing more light. Besides light absorption, other aspects such as efficient exciton dissociation, charge transportation, and charge collection are of crucial importance, and all of them are strongly influenced by the volume morphology of the photoactive layer. In this study of bulk-heterojunction solar cells based on poly(3- hexylthiophene) (P3HT) and a methanofullerene derivative (PCBM), we show that the resulting P3HT/PCBM morphology is strongly determined by the layer thickness because the kinetics of solvent evaporation and crystallization is different in films of different thickness. For the preparation conditions chosen in this study, an optimum morphological organization of the photoactive layer characterized by high crystallinity of P3HT, viz. numerous crystalline P3HT nanowires forming a genuine three-dimensional network, and enrichment of crystalline P3HT closer to the hole collecting electrode can only be achieved for relatively thin (100 nm) P3HT/PCBM layers. Corresponding devices absorb only a limited fraction of all available photons but have the highest efficiency. © 2009 American Chemical Society