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We previously demonstrated that coagulation factor VIII (FVIII) accelerates proteolytic cleavage of von Willebrand factor (VWF) by A disintegrin and metalloprotease with thrombospondin type 1 repeats (ADAMTS13) under fluid shear stress. In this study, the structural elements of FVIII required for the rate-enhancing effect and the biological relevance of this cofactor activity are determined using a murine model. An isolated light chain of human FVIII (hFVIII-LC) increases proteolytic cleavage of VWF by ADAMTS13 under shear in a concentration-dependent manner. The maximal rate-enhancing effect of hFVIII-LC is ~8-fold, which is comparable with human full-length FVIII and B-domain deleted FVIII (hFVIII-BDD). The heavy chain (hFVIII-HC) and the light chain lacking the acidic (a3) region (hFVIII-LCΔa3) have no effect in accelerating VWF proteolysis by ADAMTS13 under the same conditions. Although recombinant hFVIII-HC and hFVIII-LCΔa3 do not detectably bind immobilized VWF, recombinant hFVIII-LC binds VWF with high affinity (KD, ~15 nm). Moreover, ultra-large VWF multimers accumulate in the plasma of fVIII−/− mice after hydrodynamic challenge but not in those reconstituted with either hFVIII-BDD or hFVIII-LC. These results suggest that the light chain of FVIII, which is not biologically active for clot formation, is sufficient for accelerating proteolytic cleavage of VWF by ADAMTS13 under fluid shear stress and (patho) physiological conditions. Our findings provide novel insight into the molecular mechanism of how FVIII regulates VWF homeostasis.
Proteolytic cleavage of ultra-large von Willebrand factor (VWF)3 on endothelial cells (1, 2) and in flowing blood (3, 4) by a plasma metalloprotease, ADAMTS13, is crucial for normal hemostasis. ADAMTS13 cleaves VWF at the specific Tyr1605–Met1606 bond in the central A2 domain (5, 6). This cleavage is dramatically accelerated by fluid shear stress (4, 6, 7) or mild denaturization with urea (5, 8) or guanidine (6, 9), which alters VWF conformation and exposes binding and cleavage sites. Inability to cleave VWF as a result of a severe deficiency of plasma ADAMTS13 leads to an accumulation of ultra-large VWF on endothelial cells or in blood (10). This triggers spontaneous platelet aggregation and disseminated microvascular thrombosis, characteristic of thrombotic thrombocytopenic purpura (11, 12). Moreover, mild to moderate deficiency of plasma ADAMTS13 has been shown to be a risk factor for cardiovascular events such as myocardial infarction (13–16) and ischemic cerebral stroke (17, 18).
In addition to fluid shear stress, we and others have shown that cleavage of soluble multimeric VWF by ADAMTS13 under fluid shear stress is dramatically accelerated by coagulation factor VIII (FVIII) (19), platelets (20, 21), and glycoprotein 1bα (21, 22). FVIII and platelets appear to synergistically accelerate cleavage of VWF by ADAMTS13 under these conditions (21). We have previously shown that the FVIII B-domain is not required, but the a3 in the context of two chain B-domainless FVIII, which binds VWF with high affinity, is required for the rate-enhancing effect on proteolytic cleavage of VWF by ADAMTS13 under shear stress (19).
Here, we show that an isolated light chain of FVIII, which is biologically inactive for clot formation, is sufficient for accelerating proteolytic cleavage of VWF by ADAMTS13 in vitro using a fluid shear-based assay and in vivo using fVIII−/− mice expressing FVIII variants via a hydrodynamic approach. This rate-enhancing effect by FVIII light chain also depends on its high affinity binding with VWF; a light chain of FVIII lacking the a3 region and a heavy chain of FVIII, which do not bind VWF detectably, exhibit no effect on ADAMTS13-mediated VWF proteolysis under the same conditions. Our findings may shed more light on the structure-function relationship of FVIII in regulation of the VWF-ADAMTS13 axis, which helps in understanding the clinical heterogeneity of patients with severe hemophilia A.
The cDNA fragments encoding a heavy chain (hFVIII-HC), a light chain (hFVIII-LC), and a light chain lacking a3 (hFVIII-LCΔa3) of human FVIII were amplified by PCR using hFVIII-BDD as a template (19). The primer pairs were as follows: hFVIII-HC (5′-GAGTACTCCCTCTCAAAAGCGGGCATG-3′ and 5′-GGAGAAGCTTCTTGGTTCAAT-3′); hFVIII-LC (5′-GAAATAACTCGTACTACTCTT-3′ and 5′-GTAGAGGTCCTGTGCCTCGCA-3′); and hFVIII-LCΔa3 (5′-AGCTTTCAAAAGAAAACACGA-3′ and 5′-GTAGAGTCCTGTGCCTCGCA-3′). The amplified hFVIII-LC and hFVIII-LCΔa3 fragments were cloned into pSecTag/FRT/V5-His TOPO vector (Invitrogen). An hFVIII-HC fragment was cloned into pcDNA3.1 V5-His TOPO vector (Invitrogen) according to the manufacturer's recommendation. All three constructs were tagged at their C termini with a V5-His epitope (Fig. 1A). Plasmids were sequenced at the Nucleic Acid Core Facility at the Children's Hospital of Philadelphia to confirm the accuracy. Plasmids encoding a B-domain deleted human factor VIII (hFVIII-BDD) (19) or canine FVIII (cFVIII-BDD) (23) were described in other studies.
Baby hamster kidney cells were used to express human recombinant hFVIII-BDD and cFVIII-BDD, as described previously (19, 23, 26). Human embryonic kidney (HEK293) cells were used to express recombinant hFVIII-LC, hFVIII-LCΔa3, and hFVIII-HC. Cells were grown in Dulbecco's modified Eagle's medium (DMEM) (Invitrogen) containing 10% of FetalPlex (Gemini BioProducts, West Sacramento, CA). The cells were transfected with LipofectAMINE2000 and plasmid DNA in serum-reduced Opti-MEM. For those vectors without the G418-resistent gene, co-transfection with pcDNA3.1 (10:1 ratio) containing the neomycin resistance gene (Invitrogen) was carried out. Stable clones were selected after culturing cells for 10 days in the presence of 0.5–1.0 mg/ml of G418 (Invitrogen). The positive clones were identified by Western blotting and immunofluorescent staining with anti-FVIII IgG (ESH-8) that recognizes light chain or anti-V5 IgG (Invitrogen) for V5-His-tagged proteins as described previously (25).
Serum-free condition medium of stable cell lines expressing recombinant FVIII or variants was collected daily for a total of 2.5–10 liters. Recombinant hFVIII-BDD and cFVIII-BDD were purified and quantified as described previously (19, 23, 26). Recombinant hFVIII-HC without the C-terminal epitope was kindly provided by Dr. Philip Fay at the University of Rochester, New York. Recombinant hFVIII-LC, hFVIII-LCΔa3, and hFVIII-HC were purified by SP-Sepharose ion exchange chromatography, followed by a nickel-nitrilotriacetic acid affinity column as described previously (19).
The purity and integrity of the purified recombinant FVIII variants were determined by 10% SDS-polyacrylamide gel with Coomassie Blue staining. All purified recombinant FVIII variants except for hFVIII-HC were quantified by absorbance at 280 nm corrected with light scattering at 320 nm. Partially purified hFVIII-HC (V5-His tagged) was quantitated by Western blotting with monoclonal anti-V5 IgG using a positopeTM as a reference (Invitrogen) (25).
Human VWF was purified from plasma by cryoprecipitation followed by gel filtration on a Sephacryl-300 column (2.5 × 100 cm) (GE Healthcare) as described previously (21). Recombinant human ADAMTS13 (V5-His tagged) was expressed from stably transfected HEK293 cells and purified by Q-fast flow ion exchange followed by a nickel-affinity chromatography as described previously (7, 21).
Purified plasma VWF (37.5 μg/ml) was incubated in a PCR tube (Fisher Scientific, Newark, DE) for 10 min with 50 nm recombinant human ADAMTS13 in the absence or in the presence of FVIII variants at various concentrations in 20 mm HEPES, 0.15 m NaCl, 5 mm CaCl2, and 1.0 mg/ml BSA, pH 7.5. The reaction mixture (20 μl) was subjected to constant vortexing at 2,500 rpm for 10 min at room temperature as described previously (7, 19, 21). The reaction was quenched by boiling the sample with an equal volume of sample buffer (125 mm Tris, 10% glycerol, 2% SDS, and 0.01% bromphenol blue, pH 6.8) for 5 min. A fraction of sample was fractionated on a 5% Tris-glycine SDS-polyacrylamide gel. Alternatively, samples were denatured at 60 °C for 20 min with sample buffer (60 μl) (70 mm Tris, pH 6.8, 2.4% SDS, 0.67 m urea, and 4 mm EDTA, 10% glycerol, 0.01% bromphenol blue). Denatured sample (10 μl, ~90 ng of VWF) was fractionated on a mini-gel containing 1% agarose (Lonza, Rockland, ME). After being transferred onto a nitrocellulose membrane (Bio-Rad), VWF cleavage product or multimers were determined by Western blotting. Membrane was blocked with 1% casein in TBST for 30 min and incubated with rabbit anti-VWF IgG (Dako, Carpinteria, CA) (1:5,000) in 1% casein/TBST, followed by IRDye 800CW-labeled goat anti-rabbit IgG (LI-COR Bioscience, Lincoln, NE) (1:20,000) in the same buffer. When resolved by SDS-PAGE, the cleavage product at 350 K was quantified by densitometry using ImageJ software as described previously (19, 21). When resolved by agarose gel electrophoresis, the ratio of the low molecular weight bottom band to the high molecular weight bands was quantified in a similar manner.
A microtiter plate was coated with 100 μl of human VWF (2 μg/ml) and blocked with 1% casein in PBS, pH 7.4. Recombinant FVIII variants at various concentrations diluted with PBS containing 0.2% casein were added and incubated at 25 °C for 1 h. After being washed with PBS, bound FVIII variants were detected by peroxidase-conjugated mouse anti-human monoclonal anti-FVIII IgG (ESH-8HR) (1:3,000) that recognizes the C2 domain in the light chain of FVIII (American Diagnostica Inc., Stamford, CT) or peroxidase-conjugated monoclonal anti-V5 (1:1,000) (Invitrogen). A pre-mixed chromogenic substrate, 3,3′,5,5′-tetramethylbenzidine (TMB) (Thermo Fisher Scientific, Rockford, IL), was added for color reaction. The absorbance was determined at 450 nm in a SpectroMax microtiter plate reader (Molecular Device, Sunnyvale, CA).
The Institutional Animal Care and Use Committee (IACUC) at the University of Pennsylvania and the Children's Hospital of Philadelphia approved the protocol for the mouse study. FVIII-deficient mice (fVIII−/−) in a C57BL6/129 strain with exon-16 deletion were described previously (27). Mice at the age of 6–8 weeks were injected with 2 ml of saline alone or saline containing 100 μg of plasmid DNA (endotoxin-free) via a tail vein within 5 s as described previously (28). 48 h after injection, whole blood (200 μl) was collected after tail clip and anti-coagulated with sodium citrate (3.8%). Platelet-poor plasma was obtained after centrifugation at 10,000 rpm for 10 min and stored in small aliquots at −80 °C until use.
Plasma antigen levels of FVIII variants expressed after hydrodynamic injection were determined by a home grown ELISA. A microtiter plate was coated with polyclonal goat anti-FVIII IgG (1:2,000) at 25 °C for 1 h. The plate was blocked with 1% (w/v) casein in PBST for 30 min. Purified recombinant hFVIII-BDD or cFVIII-BDD (0, 5, 10, 20, 40, 80, 160, and 320 ng/ml) or mouse plasma (1:20 or 1:40) in PBS was incubated in antibody-coated wells for 2 h. After being washed with PBS, bound FVIII variants (hFVIII-BDD, cFVIII-BDD, hFVIII-LC, and hFVIII-LCΔa3) were detected by peroxidase-conjugated monoclonal anti-FVIII IgG (ESH-8HR) (1:1,000) (American Diagnostica, Stamford, CT) for 1 h. For quantification of plasma hFVIII-HC antigen, mouse plasma was incubated in a nickel-coated microtiter plate for 2 h. The bound hFVIII-HC was detected by monoclonal anti-V5 IgG (Invitrogen), followed by a horseradish peroxidase-conjugated rabbit anti-mouse IgG (DAKO, Carpinteria, CA). A pre-mixed TMB solution was added for color development, and absorbance at 450 nm was determined by a SpectroMax microtiter plate reader as described above.
Plasma VWF antigen was quantified by an ELISA as described previously (29). A microtiter plate was coated with rabbit anti-vWF IgG (Dako, Carpinteria, CA) (1: 2,000) and blocked with 1% casein in PBS containing 0.05% Tween 20 (TBST). Mouse plasma diluted (1:20 and 1:40) with 0.2% casein in TBST was added and incubated for 1 h. After being washed with PBS, bound VWF was detected by peroxidase-conjugated rabbit anti-VWF IgG (1:2,000) (Dako, Carpinteria, CA). Pooled murine plasma from C57BL6 was used as a reference. A pre-mixed TMB solution was used for color development. The absorbance at 450 nm was determined with a SpectroMax microtiter plate reader (Molecular Device, Sunnyvale, CA).
Mouse plasma (1 μl) was denatured at 60 °C for 20 min with 10 μl of sample buffer (70 mm Tris, pH 6.8, 2.4% SDS, 0.67 m urea, and 4 mm EDTA, 10% glycerol, 0.01% bromphenol blue). The proteins were fractionated on a 1% agarose gel (7 × 8 cm) on ice. After being transferred to a nitrocellular membrane, VWF multimers were detected by incubation with rabbit anti-VWF IgG (1:5,000) (Dako, Carpinteria, CA), followed by IRDye 800CW-labeled goat anti-rabbit IgG (1:20,000) in 1% casein/TBST. Odyssey imaging analysis (LI-COR Bioscience, Lincoln, NE) was used to scan the membrane (19, 30). Densitometry analysis using ImageJ software determined the formation of cleavage product or the ratio of high to low molecular weight VWF multimers.
The difference in means between the control and experimental groups was determined by one-way analysis of variance using Minitab16 software. p values less than 0.05 and 0.01 are considered to be statistically significant and highly significant, respectively.
We previously found that full-length FVIII accelerates proteolytic cleavage of VWF by ADAMTS13 under fluid shear stress (19). However, the domain components of FVIII required for the cofactor activity to enhance VWF proteolysis are not fully understood. We therefore prepared various recombinant FVIII variants, including hFVIII-HC, hFVIII-LC, and hFVIII-LCΔa3, in addition to hFVIII-BDD and cFVIII-BDD (Fig. 1A). All variants except for hFVIII-BDD and cFVIII-BDD contained a V5-His epitope at their C-terminal end to facilitate purification and detection (Fig. 1A). All variants except for hFVIII-HC were purified to homogeneity as demonstrated by SDS-PAGE and Coomassie Blue staining (Fig. 1B). The hFVIII-HC was only partially purified due to low secretion of this variant from stably transfected cells (data not shown). Therefore, a purified preparation of hFVIII-HC (without V5-His) was obtained from Dr. Phillip J. Fay, University of Rochester, School of Medicine and Dentistry, Rochester, NY.
As shown, cFVIII-BDD was purified as a single chain protein (Mr ~160,000) (Fig. 1B, 1st lane), whereas hFVIII-BDD was purified as a two-chain protein (Mr ~90,000 and ~80,000) (Fig. 1B, 2nd lane). The hFVIII-HC and hFVIII-LC were secreted and purified as a single chain protein with Mr of ~95,000 and ~84,000, respectively (Fig. 1, B, 3rd and 4th lanes, and C, 6th and 7th lanes). Interestingly, purified hFVIII-LCΔa3 exhibited two different sizes (~80,000 and ~75,000) (Fig. 1B, 5th lane), both of which were recognized by monoclonal anti-V5 IgG on a Western blot (Fig. 1C, 8th lane), suggesting potentially aberrant N-terminal processing of hFVIII-LCΔa3 during biosynthesis.
Moreover, using a one-stage clotting assay, we had previously shown that hFVIII-BDD exhibited specific clotting activity, comparable with full-length FVIII (19), whereas cFVIII-BDD exhibited an ~3-fold increase in specific activity compared with hFVIII-BDD (23). No clotting activity was detected with hFVIII-LC, hFVIII-HC, and hFVIII-LCΔa3 (data not shown).
To determine whether the isolated light chain of FVIII, a biologically inactive FVIII variant for clot formation, accelerates VWF proteolysis by ADAMTS13, we incubated a fixed concentration of VWF (150 nm) with ADAMTS13 (50 nm) in the presence of various concentrations of recombinant hFVIII-LC (0, 0.5, 1.0, 2.5, 5.0, and 10 nm) for 10 min under constant vortex at the rotation rate of 2,500 rpm. The proteolytic cleavage of VWF was determined by SDS-PAGE and Western blotting as described under “Experimental Procedures.” Recombinant hFVIII-LC increased the formation of the proteolytic cleavage product (~350,000, a dimer of C-terminal fragments) in a concentration-dependent manner (Fig. 2, A and B). The maximal rate-enhancing effect was ~8-fold (Fig. 2B). Similar fold of rate-enhancing effect by hFVIII-LC without a V5-His epitope on the cleavage of VWF by ADAMTS13 under the same conditions was observed (data not shown). The concentration of hFVIII-LC achieving 50% of the maximal enhancing effect (C50) was ~1.0 nm (Fig. 2B), quite similar to that of full-length FVIII and hFVIII-BDD that we previously reported (19). Addition of EDTA (15 mm) into the reaction completely inhibited proteolytic cleavage of VWF by ADAMTS13 even in the presence of 10 nm hFVIII-LC (Fig. 2, A, 2nd lane, and B). These results demonstrate for the first time that the isolated light chain of FVIII is sufficient for accelerating proteolytic cleavage of VWF by ADAMTS13 under fluid shear stress.
To determine whether the light chain of FVIII or the heavy chain, both lacking the high affinity binding site for VWF, was able to enhance VWF proteolysis by ADAMTS13 under the same conditions, a fixed concentration of human VWF (150 nm) and human ADAMTS13 (50 nm) was incubated with various concentrations of recombinant hFVIII-LCΔa3 or hFVIII-HC (0, 0.5, 1.0, 2.5, 5.0, and 10 nm) for 10 min under constant vortexing at a rotation rate of 2,500 rpm. The proteolytic cleavage of VWF was determined by SDS-polyacrylamide gel and Western blotting. As shown, recombinant hFVIII-LCΔa3 (Fig. 2, C and D) or hFVIII-HC (Fig. 2, E and F) at any given concentration (up to 10 nm) did not exhibit a rate-enhancing effect on cleavage of VWF by ADAMTS13 under the same conditions. Recombinant hFVIII-BDD (20 nm) used as a positive control dramatically increased the formation of the proteolytic cleavage product (Fig. 2, 1st lane). These results suggest that the acidic a3 region in the light chain of FVIII is required for accelerating VWF proteolysis by ADAMTS13 under shear stress.
To assess the binding affinity between FVIII variants and VWF, increasing concentrations of recombinant cFVIII-BDD, hFVIII-BDD, hFVIII-LC, hFVIII-HC, and hFVIII-LCΔa3 (0, 1.56, 3.1, 6.25, 12.5, 25, 50, and 75 nm) were incubated with human VWF immobilized on a microtiter plate (1.0 μg/well). The bound FVIII variants were determined by anti-FVIII IgG or anti-V5 IgG (if V5-His tagged) as described under “Experimental Procedures.” We showed that cFVIII-BDD (Fig. 3A), hFVIII-BDD (Fig. 3B), and hFVIII-LC (Fig. 3C) bound human VWF in a concentration-dependent manner. The dissociation constants (KD) for cFVIII-BDD, hFVIII-BDD, and hFVIII-LC binding to immobilized human VWF were ~1.1, ~1.3, and ~15.6 nm, respectively (Table 1). These results indicate that the heavy chain may contribute to the overall binding affinity of FVIII to immobilized VWF. Moreover, the KD values for cFVIII-BDD, hFVIII-BDD, and hFVIII-LC to bind immobilized murine VWF was 1.9, 1.7, and 26 nm, respectively. These results suggest that there is little species difference between VWF and FVIII binding. As predicted, no detectable binding was observed between hFVIII-LCΔa3 (or hFVIII-HC) and immobilized human VWF and murine VWF under the same conditions (data not shown). These results indicate that the light chain, particularly the a3 region in the light chain of FVIII, contains the major binding site for VWF.
Although full-length FVIII, B-domainless FVIII variant, and the isolated light chain of FVIII are able to accelerate proteolytic cleavage of VWF by ADAMTS13 under mechanically induced shear stress in vitro, the physiological relevance of such an enhancing effect has never not been determined in vivo. Using a hydrodynamic approach, we were able to reconstitute plasma FVIII in fVIII−/− mice with various FVIII variants, including cFVIII-BDD, hFVIII-BDD, hFVIII-LC, and hFVIII-LCΔa3 at levels between 0.7 and 2.8 μg/ml measured 48 h after injection (Table 2). Plasma VWF antigen and multimer distribution in fVIII−/− mice were determined after 48 h of post-reconstitution. We showed that although plasma VWF antigen did not change dramatically in different groups of mice, the ratios of high to low molecular weight VWF multimers were significantly higher in fVIII−/− mice receiving saline alone (1.7 ± 0.6, n = 10) than those receiving plasmids encoding cFVIII-BDD (0.4 ± 0.4, n = 16) (p < 0.01), hFVIII-BDD (0.9 ± 0.2, n = 9) (p < 0.01), hFVIII-LC+HL (0.4 ± 0.2, n = 5) (p < 0.01), and hFVIII-LC (1.0 ± 0.3, n = 9) (p < 0.01) (Fig. 4 and Table 2). In contrast, the ratios of high to low molecular weight) VWF multimers in the fVIII−/− mice receiving plasmids encoding hFVIII-LCΔa3 (1.4 ± 0.4, n = 11) and hFVIII-HC (not shown) were not significantly different from those receiving saline alone (Fig. 4 and Table 2). These results suggest that the isolated light chain of FVIII is required and sufficient for accelerating VWF proteolysis by ADAMTS13 under (patho) physiological conditions, thus altering the distribution of VWF multimers in circulation. However, the isolated heavy chain or light chain of FVIII lacking the a3 region has no enhancing effect in VWF proteolysis under the same conditions.
In this study, we demonstrate both in vitro and in vivo that an isolated light chain of FVIII, which is biologically inactive for enhancing clot formation, appears to be sufficient for accelerating proteolytic cleavage of VWF by ADAMTS13. The maximal rate enhancing effect with 5 nm of hFVIII-LC was ~8-fold, comparable with that of hFVIII-BDD (Fig. 2, A and B). The C50 is estimated to be ~1.0 nm (Fig. 2, A and B), which is within the physiological ranges of FVIII in human plasma. In contrast, a light chain of FVIII lacking the acidic a3 region (hFVIII-LCΔa3) and hFVIII-HC, both of which do not bind VWF, exhibit no enhancing effect under the same conditions (Fig. 2, C–F). These results indicate that high affinity binding of FVIII to VWF through the acidic a3 region may be critical for accelerating VWF proteolysis.
However, the rate-enhancing effect of FVIII on VWF proteolysis does not appear to be in a linear relationship with its VWF binding affinity. For instance, although hFVIII-LC binds human VWF (KD = 15 nm) and murine VWF (KD = 26 nm) ~10 times less than binding of hFVIII-BDD and full-length FVIII to human VWF (KD = 1.3 nm) and murine VWF (KD = 1.9 nm) (Table 1), the rate-enhancing activity is quite similar to that of hFVIII-BDD (Fig. 2) and full-length FVIII (19).
The biological relevance of FVIII and its variants on regulating VWF proteolysis by ADAMTS13 was further assessed using a murine model. At the steady state, plasma levels of VWF antigen in fVIII−/− mice are increased by ~2-fold when compared with wild-type mice in the same genetic background C57BL6/129 (data not shown), whereas the ratio of high to low molecular weight VWF multimers was not statistically different between the two groups of mice. Similar increases in plasma levels of VWF antigen and ristocetin-cofactor activity were observed in patients with severe hemophilia A compared with healthy controls (32). These findings suggest that FVIII may be involved in regulating VWF homeostasis under physiological conditions. However, the interpretation of these data may be complicated by the fact that plasma VWF antigen levels differ significantly among various strains of mice or human individuals.
Although hydrodynamic challenge is considered to be a nonphysiological transfection method, it activates endothelial cells and triggers the release of ultra-large VWF from endothelial cells, resulting in an accumulation of ultra-large VWF multimers in plasma of the fVIII−/− mice receiving saline alone. However, these ultra-large VWF multimers were not observed in the same mice receiving plasmids encoding cFVIII-BDD, hFVIII-BDD, and hFVIII-LC or hFVIII-LC+HC (Fig. 4 and Table 2). These results demonstrate for the first time that reconstitution of functional and nonfunctional FVIII variants restores the distribution of plasma VWF multimers in severe hemophilia A mice.
Consistent with the in vitro data, reconstitution in the fVIII−/− mice with a plasmid encoding hFVIII-LCΔa3 (Fig. 4 and Table 2) has no effect on plasma VWF multimer distribution. However, the in vivo effect of hFVIII-HC alone remains to be determined because plasma levels of the expressed hFVIII-HC were low, only 1/10 of the plasma levels of other constructs after the hydrodynamic injection (data not shown). The low expression of the heavy chain is due to inefficient secretion of this chain when it is expressed alone (24, 26). Interestingly, co-expression of hFVIII-LC dramatically improves the secretion of the heavy chain, thereby synergistically enhancing proteolytic cleavage of VWF by ADAMTS13 in vivo. The efficacy of the co-expressed hFVIII-LC and hFVIII-HC appears to be similar to that of hFVIII-BDD but better than that of hFVIII-LC alone (Table 2), suggesting that the heavy chain is able to stabilize the light chain to improve the light chain function.
The implication of these findings is not clear. Patients with severe hemophilia A (with FVIII activity <1%) are heterogeneous in their clinical presentations (31). Further investigation of plasma VWF multimer distribution in correlation with the genetic basis that results in severe FVIII deficiency may shed new light on how FVIII-dependent proteolysis of VWF may play a role in the modification of the clinical phenotype in patients with severe hemophilia A.
In conclusion, our findings provide novel insight into the structure-function relationship of FVIII in the regulation of ADAMTS13-mediated proteolysis in vitro under fluid shear stress and in vivo under (patho) physiological conditions. This may help explain the heterogeneity of plasma VWF multimer distribution and clinical phenotype in patients with hemophilia A.
We thank Philip J. Fay (Professor of Biochemistry, University of Rochester, School of Medicine and Dentistry, Rochester, NY) for providing us with purified recombinant hFVIII-HC (no tag) (data not shown). We also thank Sriram Krishnaswamy (Professor, Division of Hematology, The Children's Hospital of Philadelphia) for key reagents and helpful comments and discussion.
*This work was supported, in whole or in part, by National Institutes of Health Grants HL079027 and HL074124. This work was also supported by American Heart Association Postdoctoral Fellowship 09POST2250397 (to W. C.), Bayer Hemophilia awards, and American Heart Association Grant 0940100N (to X. L. Z.).
3The abbreviations used are:

domain: biology


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text:
PLATTSBURGH, N.Y. (WCAX) Could New York become two states? A newly proposed bill in the state Senate aims to break up the Empire State. Our Kelly O'Brien took to the streets to see what New Yorkers think about it.
Could you imagine 51 states in the USA?
"I don't think it's a wise idea," said John Voltra of Plattsburgh.
That could be New York's fate if there is enough interest in a newly proposed bill.
"What this bill proposed would do is to take a nonbinding referendum on the ballot to see if there is interest in doing it," said Assemblyman D. Billy Jones, D-Chateaugay.
So, what do folks in the North Country have to say?
"I think it should be one state, not only historically but let's not complicate things more," said William Crosby of Beekmantown.
"New York was built from New York City all the way up to here and Montreal," said John Voltra of Plattsburgh.
"That would be great," said Lucinda LaFountain of Plattsburgh. "New York City they can do away with."
Under the proposal, New York City and surrounding counties would be considered downstate. The rest of New York would be upstate. Supporters of the idea say New York is a very large and very diverse place, with many distinct cultural, economic and historical regions.
"It's very interesting," Jones said. "I know people in my district and in the North Country have a lot of ideological differences with people in New York City. There are a lot of things that affect us that affect them differently. So, I do think it's interesting."
"I think it's a little of each. Yeah, downstate has a bigger population, so more money probably ends up going there per capita. More population, area-- that may be fair. But I think upstate has been treated fairly well," Crosby said.
Others feel that the major city is hurting the North Country.
"We wouldn't if we didn't have New York City, I think we could survive," LaFountain said.
The bill will be on the ballot for voters to see how much public interest there is in the divide.
"I would say there's always a possibility," Voltra said. "Today you can't predict how people are going to vote."
One thing everyone could agree on is we need more people coming to the North Country and jobs to keep them here.
"If they could somehow bring businesses up farther north, it's just there's not work for people. We need work," said Sandra Greenwood of Fort Jackson.
The bill is currently in committee but has not made it on the floor.

domain: geography


text:
Toronto electronic music lovers get ready, cuz this Saturday January 18th, some serious sweat is in the forecast.
First time I heard Hatiras play was live at WEMF 2006 in Tweed. I was pretty new to the scene at the time and didn’t really know what was going on, except I knew that Hatiras was making me dance. I’m not gonna lie, it helped that his name was lit up in big letters because I didn’t forget it – yes I was that clueless then 😛
Have you listened to ‘Message from 04244‘ yet? Do it now. It’s actually pretty awesome. My favourite track is Weed Molly….cuz I’m a raver like that. Every track on the album is different and I predict that each one tells a different part of the space love story. What would sex be like in space? Anyway, this album is as fluid and diverse as the depths of outer space. From the funky hip-hip beats of Madly in Love to the big room dance vibes that Message from 04244 brings, Hatiras has clearly flexed a few muscles bringing this concept to life. Every track download comes with a video, of what, I don’t know, but keep an eye on Youtube or download the tracks!
Saturday January 18th at Ryze will be crazy good. A few of us are coming out of the woodwork for this, so you should too. Don’t be lame.
For $10 reduced cover before midnight, hit up Miz Megs at djmizmegs(at)gmail.com
Here’s Hatiras’ Album Reslease Party Facebook event, click for more info.
I’m back out of retirement!
I need to blog more than I need to sleep.

domain: artistic


It is the date when the UN General assembly adopted the Convention on the Rigts of the Child – the most widely ratified document on human rights in history.
On this day, the offices of UNICEF around the world organize meetings with children and teens where they discuss the main topic of the day. Every year the child's day is devoted to a different topic. In 2018, the topic of the Universal Children's Day is 'Let's stop the violence'. The works and conclusions of meetings, talks and brainstorms in 2018 will result in Children's World Manifest to be formed in early December by the main office of the UN in New York. Ukraine also marks this day with the creation of Public Children's Manifest at UNICEF Youth Hackathon featuring 60 children and teens from all around the country. Join us to celebrate the achievement of protecting the rights of the child and remind the world leaders about the problems still remaining unsolved: https://uni.cf/go-blue
The Album «MLNL» of the ANTYTILA Band Is Available to Be Bought on CD![...]
The Stadium Tour MLNL Is Postponed to the Year 2023All gigs of the stadium tour MLNL are postponed to the year 2023.[...]
Worldwide Premiere: Ed Sheeran and ANTYTILA Present a SongEd Sheeran and ANTYTILA present collaboration – a remake of the track 2Step with Taras Topolia singing in Ukrainian.[...]
MILLENNIALANTYTILA were preparing the new album feeling the irreversibility of horrible events which was aimed to inspire and support Ukrainian people, to raise Ukrainian culture in the world.[...]


BMW has been releasing a stream of limited edition cars to the Chinese market, and to keep that stream flowing smoothly, BMW just released the M3 Carbon Fiber Limited Edition for Chinese customers. The BMW M3 Carbon Fiber Limited Edition is priced at 1.23 million yuan ($189,800), and of which only 111 units will be built.
The latest BMW M3 Carbon Fiber Limited Edition will be available only in China, and as the name says, it is wrapped with carbon fiber material throughout the car. No mechanical changes are made. The BMW M3 Carbon Fiber Limited Edition is offered in white metallic finish along with contrasting black stripes that run from the front headlights to all the way to the rear end.
Other carbon fiber pieces on the exterior include a carbon fiber roof, carbon fiber front lip splitters, carbon fiber trunk spoiler and hood inlets. And to compliment the carbon fiber look, BMW added a set of gloss-black 19-inch alloy wheels.
Finally in the interior, the carbon fiber theme continues with the carbon fiber dashboard and door panels as well. And even the black leather seats feature carbon fiber patterned material.
Source: Autohome via BMWblog


text:
I selected Kirbyworks to paint the interior of my house based upon Angie’s List reviews. My agreement with Kirby was that I would specify the colors to be used, but he would determine the brand and sheen of the paint. I…
Logo Design by Kirby Hopper
Website Design by Kirby Hopper
Copyright 2012 by Kirby Hopper
Hosted by WebHostingHub
Powered by WordPress using the Kirbyworks Theme based on the Responsive Theme.
Search Engine Optimization by
Kirby Hopper using SEO PowerSuite
Yes, I can build you a website.


text:
Ground Whole Grain Corn, Soybean Mill Run, Chicken By-Product Meal, Powdered Cellulose, Corn Gluten Meal, Soybean Meal, Chicken Liver Flavor, Soybean Oil, Iron Oxide, Vitamins (L-Ascorbyl-2-Polyposphate [Source of Vitamin C], Vitamin E Supplement, Niacin, Thiamine Mononitrate, Vitamin A Supplement, Calcium Pantothenate, Biotin, Vitamin B12 Supplement, Pyridoxine Hydrochloride, Riboflavin, Folic Acid, Vitamin D3 Supplement), Iodized Salt, Potassium Chloride, Flaxseed, Choline Chloride, Vitamin E Supplement, Dicalcium Phosphate, L-Lysine, Minerals (Ferrous Sulfate, Zinc Oxide, Copper Sulfate, Manganous Oxide, Calcium Iodate, Sodium Selenite), Taurine, L-Tryptophan, L-Carnitine, Preserved with Mixed Tocopherols and Citric Acid, Beta-Carotene, Rosemary Extract.
Daily Feeding Guide: Weight of Dog (lbs [kg]), Daily Amount to Feed for Weight Maintenance, Daily Amount to Feed for Weight Loss: 5 lb (2.3 kg), 5/8 cup (60 g), 1/2 cup (50 g); 10 lb (4.5 kg), 1 cup (100 g), 3/4 cup (75 g), 20 lb (9 kg), 1 3/4 cups (175 g), 1 1/4 cups (125 g); 40 lb (18 kg), 3 cups (295 g), 2 cups (200 g); 60 lb (27 kg), 4 cups (395 g), 2 3/4 cups (270 g), 80 lb (36 kg), 5 cups (495 g), 3 1/2 cups (345 g); 100 lb (45 kg), 5 3/4 cups (570 g), 4 1/4 cups (420 g). This product is intended for weight maintenance (dogs 1-6 years). Should you choose to feed for weight loss, consult your veterinarian. These amounts are a starting point only. Your pet may need less or more food to maintain proper weight. Adjust as needed. If you are unsure, ask your veterinarian. Feeding this food for the first time? Mix increasing amounts of your pet's new food with decreasing amounts of the old food over a 7-day period. Keep fresh water available at all times. Your pet's nutritional needs change with each lifestage. Continue to give him the same precise and balanced nutrition by transitioning your dog to Hills Science Diet pet food once he reaches 7 years of age. Protect from moisture. Store in a cool, dry place.
Always refer to the actual package for the most accurate information

domain: food


Bringing Fabrics from across the globe direct to you... since 1971.
FREE UK Postage on ORDERS over £29.50
Sugar Almond crisp Metallic Floral Brocade Jacquard
Embossed metallic Jacquard Brocade in a large floral design, design is emphasised with metallic gold thread.
Ideal as a special occasion wear, we have teamed it with 2562 matt satin in Strawberry and the metallic foil design 080 37763
HLCC: Handwash 7/30' or Dry Clean
Chenille boucle twist tweed
Smart, slubbed and twisted lurex/ polyester chenille.
Multi colours of Denim/Coral/Blue on an Inky Black ground twisted with lurex threads.
Ideal as a little Chanel style jacket. Up fashion it - with some Leather Look! , or a matching bright satin etc
HLCC: 7/30' wash with care, No Tumble
Polyester Jacquard Brocade in Blackcurrant tones, Telio design 37557
Crisp taffeta style embossed Cloque Jacquard in an unusual Blackcurrant tone highlghted with a dull off-white thread, in a bold abstract floral design.
Perfect for formals or Mother of Bride/Groom
Check out Vogue 9267 which has been created using this item, available in the Blackcurrant only not as Photo which is a different shade for example only
HLCC: WashCode: 7/30'
'Teardrop' Heavy Beaded embroidered lace from Telio design 38470
SKU: 080 38470
# exclusive design to us #
An elegant and fabulously heavy weighted 'Teardrop' embroidery with a centre pearl bead.
A warm rich shade of Blackcurrant co-ordinates well with our 5822, 2562 etc.
Clover has a co-ordinate from stretch satin 5281 or liquid satin 4200
HLCC: Specialist clean
Crochet Sequin Lace from Telio design 38474
SKU: 080 38474
Lightly crochet lace with a mini sequin throughout in an 'Art Deco' influence
Luxury 'Sea Blue' Metallic Jacquard from Telio 38231
87% Polyester 13% Metallic
Embossed luxury metallic Jacquard in soft tones
Pistachio coloured Polyester Metallic Shimmer Jacquard
SKU: 066 BR26
90% Polyester 10% Metallic
'Reversible' floral shimmer Jacquard Brocade
Item has a lovely full 'bubble' texture but yet is soft and light to the touch.
co-ordinates lovely with 5251 Thyme crepe or matt duchess satin
Would make an ideal wedding outfit
WashCode: 7/40' Handwash
'Crimson Rose' Polyester Peachskin stretch crepe
95% Polyester 5% Spandex
Richly coloured floral in 'Crimson Rose' on a Navy ground.
This item is a weighted peachskin crepe with a subtle stretch.
Has a lovely textured peachskin feel, great as a dress.
HLCC: 7/30' wash with care
'Spanish Rose' embroidered tulle from Telio design 37852
SKU: 080 37852
50% Polyester 50% Viscose
Lovely embroidered, 'Spanish Rose' multi coloured embdr in Fuschia on Black Tulle
HLCC: 7/30' with care
Turquoise and Navy swirling stretch Jersey
95% Polyester 5% Spandex
Large bold abstract floral in a enterwindling design
A bold colour print, a bright Turquoise shade onto a Denim blue ground.
This is a micro jersey so has a lot of silky drape but also has a degree of background weight.
HLCC: Wash 7/30' with care
embroidered sequin lace from Telio design 37763
SKU: 080 37763
55% Polyester 45% Nylon
Miniature mini sequin on a raised silver embroidery tulle
Polyester Double scallope embroidered lace, Telio design 37765
Embroidery on a Nylon Tulle
lovely delicate looking lace with a prominent scallope edge, would make a lovely 'duster' overcoat for Bridal.
The Showroom is
currently CLOSED to
walk in customers
Mail Order is OPEN


Toplam kayıt 1, listelenen: 1-1
Quantification of the flow noise in household refrigerators
(JVE INTERNATIONAL LTD., 2014-11-01)
The flow noise in household refrigerators is quantified in this study. First, the sound pressure measurements in a quiet room using typical household refrigerators are conducted and the noise characteristics of the ...


Today we’ll be taking a quick look at the Dell Latitude 7210 2-in-1 Business Detachable Laptop. This laptop, as the name suggests is a 2-in-1 laptop and comes with a detachable keyboard. It’s designed for business use and is very similar to Microsoft’s Surface Pro 7, both in looks and features.
The Latitude 7210 powered by an Intel 10th Gen Core™ i7 quad core processor, with 16GB of LPDDR3 ram, along with a 256GB M.2 PCIe NVMe SSD. It features a 12.3-inch touchscreen IPS panel that comes with an Anti-Glare and Anti-Smudge DX glass, supporting FHD resolution of 1920×1080. For connectivity, it comes with Intel Dual Band Wi-Fi 6 AX201 2×2 802.11ax 160MHz + Bluetooth 5.1.
The laptop is extremely thin measuring just 0.37″ (9.35mm) in thickness, and super light-weight weighing in at only 1.87lbs… that’s 0.85kg. It’s perfect for the business user who travel a lot.
One of the most attractive feature of the laptop is the detachable keyboard, which features a magnetic strip long its connectors and easily locks in place for hassle-free operation. The review sample we received did not come with a stylus pen (Dell Premium Active Pen PN579X) … which will cost you an additional $89 if you need on.
As far as graphics is concerned, the Latitude 7210 still uses the Intel UHD onboard graphics which is an integral part of Intel’s 10th Gen processor. It works well for business use, but inadequate for today’s gaming. Mind you, the Latitude 7210 is really design as a business laptop.
I’m interested to see how this 2-in-1 laptop will perform during our benchmark tests. Will the Latitude 7210’s portability and sleek looks, hinder its performance in everyday business use and applications? Let’s find out …
OK, let’s dive in and start with the specifications and features of the Dell Latitude 7210 2-in-1 Business Detachable Laptop in our next page.
You can buy the Dell Latitude 7210 2-in-1 Business Detachable Laptop from various retailers including Amazon for around USD $2,100 – https://amzn.to/32mFGjB
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Memorandum of Understanding (MoU) 2016-17 between India Trade Promotion Organisation (ITPO) and Department of Commerce (DoC), Government of India was signed by Shri L.C. Goyal, CMD, India Trade Promotion Organisation and Smt. Rita Teaotia, Secretary, Commerce, Govt. of India on 18.10.2016 in New Delhi.
On this occasion, L.C. Goyal, CMD, ITPO mentioned that for the year 2016-17, the ‘Excellent’ financial target for Revenue from operations (Gross Sales) has been fixed at Rs.250 crore. He also mentioned that ITPO will make all out efforts to achieve “Excellent” rating as per the targets given by DPE for the year 2016-17 and ensure delivery of better services to all the stakeholders .


Turrialba Volcano steams
Not to be outdone by its cousin to the west, Turrialba Volcano has been steaming — literally — after the attention Arenal Volcano got for releasing smoke-like vapor in recent days. The Volcanological and Seismological Observatory of Costa Rica (OVSICORI) reported Wednesday that the same phenomenon that attracted attention to Arenal, which has been largely quiet for several years now, was at work in the crater of Turrialba Volcano.
The smoking crater is, in fact, rain water turned into steam by geothermal heat from the interior of the colossus, according to a statement from OVSICORI. Heavy rains falling on the volcano during the last several days seep into the volcano and escape out the volcano as steam in fumarole-like plumes. Turrialba is still under observation, but this recent steam does not mark a change in the volcano’s activity, experts say.
The same effect was observed at Arenal Volcano recently. Arenal is still considered active but has fallen into what scientists expect to be a long slumber. For five years, since its dramatic eruptions stopped in 2010, OVSICORI has not registered any magmatic gasses or internal seismic activity at Arenal.
Turrialba Volcano, on the other hand, has been much more active lately. A rash of ash eruptions blanketed the surrounding countryside and reached San José and beyond earlier this year. Civil aviation authorities even closed the country’s main airport, Juan Santamaría International Airport, several times because of ash in the air and on the runway.
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Lady Saints Nail Down Win Over New Mexico
KSCB News - December 1, 2014 7:13 am
They keep their composure. That’s one of the aspects of Seward’s 68-63 win over previously unbeated New Mexico Junior College which impressed Seward head coach Toby Wynn. Down 12 twice in the first half (20-8 and 25-13) and trailing 37-31 at halftime, Seward rallied for a their best win of the season dropping the Lady T-Birds to 11-1.
Brooklyn Artis led Seward with 21 incluing a clutch left wing three pointer late in the game to give Seward a 67-63 lead. Daniela Galindo scored 17 giving her 37 points in the two games at Garden City this weekend. Rebekah Hatchard added 10. Seward held the Lady T-Birds to 35 percent shooting. Seward made seven of 14 three pointers in the win. Seward is 9-0.
Seward hosts the Pizza Hut Classic Friday and Saturday night at the Green House. Seward plays Fort Carson Friday at 6 on B107.5 and Northwest Tech Saturday at 6 on 1270 and kscbnews.net.


8-22-2011Human Rights Defenders Program
Last week, Human Rights First reported that Jalila al-Salman was hospitalized after a 10 day hunger strike protesting her detention. Over the weekend, she and fellow detainee, Roula Al Saffar, were released from prison after almost five months of detention. While the release is favorable, both will face trial in the coming months. At present, three women and several dozen men are still detained for participating in anti-government activities.


Auditions for Thrillpeddlers new Paris Revue!
The theatre company that brought S.F. “Pearls Over Shanghai” and other Cockettes musicals as well as 15 years of “Shocktoberfest,” turns its sights towards Paris in a revolutionary new musical and comedic revue.
General Audition: Tuesday 12/9 from 7:00 – 9:30 pm
Alt. General & Callbacks: Monday 12/15 6:00 – 9:00 pm
Please prepare one upbeat song from (or in the style of) early to mid-20th Century repertoire. Performing a monologue and a special skill is also encouraged. Composer and musical director Scrumbly Koldewyn will provide accompaniment. Don’t have sheet music? Not to worry, Scrumbly will get you singing!
Rehearsals (7 weeks): To be scheduled on weeknights and Saturday days, 1/26/15 – 3/18/15.
Performances (30 total): Selected acts on 2/12-14, 2/28, plus two additional special events TBD, followed by an 8-week run on Thursdays, Fridays & Saturdays at 8 pm 3/12 – 5/2, 2015.
Location: Auditions, rehearsals, and performances all take place at The Hypnodrome, 575 10th St., San Francisco.
To schedule an audition contact Russell Blackwood at email@example.com or call 415-377-4202


text:
Extraordinary opening of the General Library for December examinations
from 13 November to 22 December 2006.
Monday to Friday
Weekends and public holidays
from 8:30 am to 3 pm
from 8 am to 3 pm
Students wishing to use the library after 3 am can stay at the permanent Reading Room on the first floor which is open 24 hours every day, including weekends and public holidays.
During the extraordinary opening period at weekends and public holidays, lecture rooms 'Sótano 1' and 'Sótano 2', at the Lecture Building no. 2 will remain open from 9 am to 9 pm.
Monday to Friday: free admissions to the General Library until 3 pm. From 3 pm onwards, admission is restricted to UA members, to any other University members or to those under agreement with the UA.
Weekends and public holidays: admission is restricted to UA members, to any other University members or to those under agreement with the UA for which accreditation is required. The rest of the users can use lecture rooms "Sótano I" and "Sótano 2" at the Lecture Building no. 2
Reference and bibliographical services are open during public hours.
Universidad de Alicante Carretera San Vicente s/n 03690 San Vicente del Raspeig Alicante (Spain)

domain: library


text:
For four days My brow was furrowed
Eyes full of sorrow
The energy suddenly appeared
Seeds planted and cleared
For the nighttime sun
Feeds the energy and I won.
For everything I have felt
My thoughts that were not svelte
Came and went away
On this very day
It followed me around
It didn’t make a sound
You feel it deep within your heart
Makes a perfect re-start
Not handed on a silver spoon
I conquered the dune
and the feelings I braved
Gave me a very special day
You will hear me say
I am living for today
Welcome to my blog. I have been journal writing for years. I took a course in Creative writing and found I enjoy writing short stories and poetry. I write more when I am hypomanic or manic.
I went into Rehab for Alcohol Addictions and have been sober for fourteen years.
Shortly after I quit drinking I was diagnosed with Bipolar 2, I have since been diagnosed with Bipolar 1.
I enjoy talking about my story, for people to learn or relate. I try and have a healthy lifestyle and try to use my coping strategies. Life is just too boring without Bipolar, it is a blessing but yet sometimes a curse.
I am currently working on a book and dream of publishing one day.
I am so excited to have found Wordpress.
This is my new adventure.

domain: hobby


text:
Pratyusha Banerjee is all set to be seen in glamourous avatar for the leading dance reality show on Colors
The promos featuring Pratyusha will soon go on air on Colors and, usually seen in traditional ghaghra-choli as Anandi, the actress will feature in a sexy body hugging dress in it.
She will compete with celebrity contestants from all walks of life for the title. This year’s list of participants is likely to see the likes of Sri Lankan cricketer Sanath Jayasuriya and ghazal singer Talat Aziz.
The judges’ panel will have Bollywood’s dancing diva Madhuri Dixit, filmmaker Karan Johar and choreographer-turned-director Remo D’Souza. The show is expected to go on air next month on Colors.

domain: celebrity


text:
Robots have inspired fiction for centuries, and as technology has advanced, robots have become a reality in many aspects of our lives. However, the idea of "robots" started in theater and literature, and as they've evolved in the real world, they've become even more amazing in the written word and on the stage. If you love science and engineering, or adore science fiction, these fictional robots may help you add to your baby name list.
The word “robot” was first used in Rossum’s Universal Robots, a 1920 Czech-language science fiction play written by Karel Capek. The word in that play referred to artificially-created life-forms, and this idea is a little different than the modern version of robot because they were closer to cyborgs or clones — they could be mistaken for human and could think for themselves.
Modern robots are generally mechanical in nature and guided by computer programs or electronic circuitry. While some appear capable of independent thought, true artificial intelligence is still science fiction.
The names we chose here are from science fiction film, television and literature, and may be exactly what you’re looking for as you build your baby name list.
And you'll see personalized content just for you whenever you click the My Feed .
SheKnows is making some changes!

domain: literature


text:
HONG KONG HORS CATEGORIE + CATEGORY III | Offscreen
There was a time when Hong Kong was the Hollywood of the East. At its peak in the early 1990s, the local film industry was second only to that of the United States as the world's biggest in terms of output per capita and as the largest exporter of product. Hong Kong played a key role in the spread of Asian cinema to the West, first in the 1970s with the martial arts productions of famous studios such as Shaw Brothers, followed in the 1980s and 1990s by the action, fantasy and horror films of John Woo, Tsui Hark and Ringo Lam.
But alongside the box-office successes, Hong Kong cinema maintained a remarkable level of creativity. Although it had its roots in traditional genres aimed at mass audiences, it also found room for experimentation, innovation and boundary-pushing, as much in terms of form and technique as in themes and genre tropes.
Under the heading "Hors Catégorie", we focus on Hong Kong cinema outside the mainstream, with a selection of 18unconventional, extraordinary, pioneering and even demented genre offerings attesting to a vibrant, audacious and visceral film culture.
We pay special attention to exploitation films that were awarded the Category III classification. Although Category III, introduced in 1988, is not a genre in itself, these shocking and sensational films had one characteristic in common: they defied good taste and conventional morality with their depictions of explicit sex and graphic violence. Far from scaring people off, the Category III rating came to be regarded as a guarantee of transgressive elements in the Hong Kong films that flooded cinemas across Asia, while remaining below the radar of most Western audiences.
We augment and enhance our selection of nine Category III films such as Naked Killer, Taxi Hunter and Sex and Zen with nine "Hors Catégorie" curios such as Encounter of the Spooky Kind, Zen Kwan Do Strikes in Paris and The Wicked City, adding up to an unforgettable feast of cinema at its most extreme, inventive and and intoxicating.
Anthony Wong, fearless as ever, plays a horny sleazebag who catches ebola from a woman he rapes in Africa. Pausing only to murder his boss and turn the corpse into burgers, he carries the virus back to Hong Kong and runs amok. Yau’s gory yuckfest is tasteless, hilarious and guaranteed to offend just about everyone.
An intrepid writer suspects an odd couple and their cat are extra-terrestrials, though he's slow to realise what we already know: these aliens are the good guys. The real threat comes from a psionic blob-monster in this sci-fi fantasy packed with demented special effects, including a crazy cat vs dog kung fu fight.
Screen goddesses Maggie Cheung, Michelle Yeoh and Anita Mui play rival superheroines who join forces to foil an evil being who has been kidnapping newborn babies in this action fantasy with wuxia-inspired effects, unexpected infant mortality and gratuitous urination. Watch out for Anthony Wong's flying guillotine!
A scholar is obsessed with seducing women, leading to slapstick comedy, tragic irony and plenty of softcore lubriciousness in this sensuous adaptation of a 17th century Chinese erotic novel. Highlights include a horse penis transplant, letter-writing by vagina, and a flute co-opted as a lesbian sex-aid.
The 1990s were a notorious period in the history Hong Kong cinema. By turns gory sexualised, violent and just plain outrageous, Category III films quickly gained a reputation as cult films. As often in extreme exploitation movies, these films challenged a whole range of societal norms and French critic Julien Sévéon will explore the taboo breaking aspects of Category III films that has made them such a cultish draw for so many. Examining films before and after the handover of Hong Kong to China, Dr Calum Waddell (University of Lincoln) will interrogate the ways in which these films seem to foreshadow the current political crisis in Hong Kong. We will also be joined by Dr Victor Fan(King’s College, London) who will turn his wide-ranging expertise in Chinese cinema and politics on to the crazy, bizarre and confrontational world of Category III.
This documentary tells you everything you need to know about Hong Kong cinema at its most extreme, from early shockfests to the shadow of real-life political events to the post-handover scene. Among the talking heads are Anthony Wong, star of some of Cat III's most outrageous hits, and Josie Ho, of Dream Home fame.
Leslie Cheung plays a struggling film director who has to compromise his ideals by making softcore porn. This sex-comedy meta-masterpiece pokes fun at Category III and features unusually nuanced female characters, notably Karen Mok as the director’s girlfriend and The Assassin's Shu Qi as his starlet, “Miss Mango”.
Hong Kong superstars Chow Yun-Fat, Simon Yam and Anthony Wong play scumbags who double-cross each other and shoot, stab or blow up anything that moves, including innocent bystanders. Early use of "Bullet Time" seals the deal in the sort of raunchy, ultra-violent shindig that makes Hollywood action pics look anaemic.
When his pregnant wife gets her dress caught in a cab door and is accidentally dragged to her death, a mild-mannered insurance salesman (Anthony Wong) flips out and wreaks revenge on bad taxi drivers. Vigilante madness, peppered with dark humour and references to Taxi Driver.
Kitty, an air hostess who shoots men in the balls, is mentored by a female assassin who keeps rapists chained up in her basement for killing practice, but starts to lose her edge when she falls for a traumatised cop (Simon Yam). A sexy, action-packed work of Cat III genius, featuring death by poisoned lipstick!
The film for which the Category III rating was first created is a gruelling account of horrific experiments conducted by Japanese scientists on Chinese prisoners during WW2. Mutilation of real cadavers, human vivisection and animal cruelty are just some of the atrocities on show. Warning: not for the squeamish.
Humans and demons coexist uneasily in this live-action adaptation of a Japanese anime. Two cops investigate a deadly new drug which turns its users into vapour, while demons jostle for power in an orgy of mad special effects, turning themselves into a spider-woman, drinking water, and a fuckable pinball machine.
Supernatural action comedy directed and choreographed by the legendary Hung, who plays a village pedicab driver. His wife's lover plots to murder him with black magic, leading to slapstick encounters with ghosts, zombies and hopping vampires, a duel between rival sorcerors and some sensational fight sequences.
Hark's epic masterpiece is a breathless parade of dazzling beauty and groundbreaking special effects. In fifth century China, a young army deserter tags along on a wuxia quest for twin magic swords to defeat an evil sect. Sammo Hung plays a wizard who keeps a Blood Demon at bay with nothing more than his eyebrows!
Leslie Cheung plays a poor debt collector who spends the night in a haunted temple and falls in love with a beautiful ghost. Alas, a long-tongued tree demon is forcing her to lure men to their doom. Can our hero escape a grisly fate? Delightful period rom-com with freaky special effects and wuxia wire-fu action.
- 1 of 2


and Whoopi Goldberg could join "The View"
The latest reports point
towards Whoopi Goldberg and Sherri Shepherd as regular hosts on
to the Los Angeles Times, the ABC show "The View" is about to
close the deal with Sherri Shepherd and Whoopi Goldberg as the
new hosts. The View has been searching for regular replacements
of Rosie O'Donnell and the latest reports indicate that Shepherd
and Goldberg may take the job.
Whoopi Goldberg would give The View a big name to replace
O'Donnell, and her versatile career would ensure that feuds
would not occur during the show. Sherri Shepherd has appeared on
The View more than dozen times, five times in July alone, a
clear indicator that the producers are seriously considering the
The two stars would also give The View an
ethnic diversity, which was lost after Star Jones Reynolds left
the daytime show.
The online betting site
Bodog Sports offered odds on
the replacement of Rosie O'Donnell. Unfortunately, neither
Whoopi Goldberg nor Sherri Shepherd were listed as contenders,
and the bet on the next View host would be a bust, if either of
them gets the job.
Published on 07/27/2007


text:
A Stribild lawsuit is a mass tort claim against Gilead Sciences, Inc., the HIV medication’s manufacturer. Stribild contains the drug TDF, which can cause serious side effects such as kidney failure and bone fractures as well as lactic acidosis.
However, Gilead failed to warn people about these risks on the TDF drug’s warning label. Moreover, there is growing evidence that Gilead covered up a safer drug in order to maximize its profits. Stribild and other 2022 TDF lawsuits are seeking compensation for:
- Medical bills to treat these side effects,
- Anticipated medical expenses for ongoing medical care,
- Pain and suffering,
- Loss of consortium suffered by the victim’s family, and
- Professional setbacks like lost wages and reduced earning capacity.
The statute of limitations for filing a claim varies by state but is generally is two years from the injury date.
In this article, our Stribild lawyers will discuss:
- 1. What is Stribild?
- 2. What are the side effects?
- 3. Are there lawsuits pending against the manufacturer?
- 4. What compensation can victims obtain?
- 5. Is there litigation against Gilead’s other HIV drugs?
1. What is Stribild?
Stribild is a brand name for an HIV treatment antiretroviral pill. This pill contains a mixture of four drugs:
- Emtricitabine, and
- Tenofovir disoproxil fumarate, or TDF.
The TDF works with the elvitegravir and the emtricitabine to prevent HIV cells already in the body from multiplying and spreading. The cobicistat in Stribild prevents the body from metabolizing the elvitegravir in Stribild.
Because Stribild only prevents HIV cells from spreading, it is not a cure for HIV. Rather, it is a treatment that needs to be continued for as long as a patient is at risk of developing HIV. Many of the people who are on Stribild work with HIV-infected patients or are in intimate relationships with people who are HIV-positive or who have AIDS.
However, the TDF medication used in Stribild is very toxic. And has to be taken in high doses because not much of it is absorbed in the body.
2. What are the side effects?
Stribild comes with a host of common side effects, including:
- Skin discoloration and rash,
- Muscle pain,
- Vivid dreams and hallucinations,
- Stomach pain and discomfort,
- Breathing problems, and
Stribild has also been known to cause some far more severe side effects, as well:
- Kidney problems, including acute kidney injury and acute renal failure,1
- Lactic acidosis, or the accumulation of lactic acid in the blood,2 and
- A decrease in bone mineral density that can lead to osteopenia, osteoporosis, broken bones, and other bone injuries and bone problems.3
Each of these severe side effects can post a significant impact on an HIV patient’s quality of life. In some cases, they can even be fatal.
2.1. Kidney damage and renal failure caused by Stribild
Most of the TDF in Stribild does not get absorbed in the body. Instead, it passes through the kidneys on its way out of the organ. The toxicity of the drug can cause substantial kidney damage. Because Stribild is a long-term treatment for HIV rather than a cure, the kidney damage it causes builds up over time.
One study monitored the kidney health of 10,841 people in the San Francisco area with HIV. All of them were taking a TDF drug to treat their condition. One thing the study monitored was whether there were abnormal protein levels in their urine – a telltale sign of Fanconi Syndrome (where impaired kidney tubule function affects the urine’s chemical makeup – which can then lead to kidney damage).
After controlling for factors like age and health, the study found that there is a 34 percent increase in signs of kidney damage for every year the patient was taking a TDF drug. Additionally, there was a 33 percent increase in developing chronic kidney disease and an 11 percent increase in a sudden and rapid loss of kidney function.4
As the kidney damage gets worse, it can lead to renal failure – a potentially lethal medical condition.
2.2. From kidney failure to lactic acidosis
One of the worst complications of kidney failure caused by Stribild is lactic acidosis.
Because the kidneys are responsible for controlling the body’s lactic acid, reduced kidney performance can lead to lactate accumulating in the blood. If enough lactic acid builds up, it can alter the acidity of the entire bloodstream, lowering its pH level. As the blood becomes more acidic, it can cause medical complications throughout the rest of the body. Some of them, like organ failure, can be fatal.
2.3. Stribild can cause a reduction in bone density
The toxicity of the TDF in Stribild is also a problem for a patient’s bone structure. When combined with the usual effects that the HIV virus has on a victim’s bone density, Stribild can lower bone mass significantly. Because Stribild is a permanent treatment, the effect that the TDF has on a victim’s bone density accumulates over time.
As the bone density decreases, the risks of breaking or fracturing a bone increase accordingly. Stribild can lower bone mass so much that the density could fall below the level of osteoporosis, a medical condition typically seen only in the elderly.
3. Are there lawsuits pending against the manufacturer?
Thousands of lawsuits have been filed against Gilead Sciences, Inc. for the side effects of Stribild and other TDF meds. These lawsuits claim that victims are entitled to compensation because Gilead did two things:
- They developed a safer drug than Stribild, but kept it off the market to maximize company profits, and
- They failed to warn doctors and patients about the risks of Stribild.
Some victims in California have joined together to file a class action lawsuit against Gilead.5 But most 2022 Complera lawsuits are proceeding through the court system individually.6 Because these lawsuits involve victims with similar injuries, they are presently being consolidated into an MDL (multidistrict litigation) out of the Northern District of California (federal court).
An MDL is where similar lawsuits are joined together to expedite litigation and a settlement. The case name is In Re. Tenofovir Disoproxil Fumarate Products Liability Litigation, MDL 2881.
Eventually, the judge will allow a few cases to go to trial, called bellwether trials. How they turn out will set the stage for negotiations with the remaining thousands of plaintiffs. The first bellwether trial is scheduled for June of 2022.
3.1. Gilead withheld a safer alternative to Stribild
Gilead first began releasing its TDF-based HIV treatment pills in 2001.7 According to the lawsuits, Gilead was already developing another drug, tenofovir alafenamide fumarate (TAF), at that point. Research into TAF showed that TAF was absorbed at higher rates in the body than TDF. This meant TAF had the same effect as TDF but at much lower doses, reducing the dangers of its toxicity.8
But Gilead’s TDF-based drugs were just being released and still had years of market protection from generic alternatives. They were also dominating the HIV treatment market. Gilead realized that they could continue to make billions of dollars each year on their TDF drugs until their market protection expired, and then release its TAF-based drugs to get a whole new period of market protection from generics. In 2004, Gilead patented its formula for TAF, announced that it would stop the development of TAF, and then hid all of the medical studies that it had conducted about the drug.9
Then in 2010 when the market protections for TDF-based HIV drugs were expiring, Gilead released the medical studies that it had done years earlier showing that TDF could cause osteoporosis, broken bones, kidney damage and failure, and lactic acidosis – making it a very dangerous drug to take. But its new breakthrough TAF drugs drastically reduced those risks, so patients should switch to the new drug right away.10
The lawsuits claim that Gilead should be held accountable for injuries suffered by people taking Stribild during the time between Gilead’s development of TAF-drugs and its release of them to the public.
3.2. Gilead failed to warn people of the dangers of taking Stribild
Drug companies have a legal duty to disclose the known risks and side effects of their products. The law requires these disclosures because it understands that doctors need them to write prescriptions and because patients use them to make informed healthcare decisions.
Victims filing Stribild lawsuits claim that Gilead failed to uphold this legal duty to warn. They claim that Gilead knew of the risks of Stribild and other TDF-based HIV drugs, and point to much of the same evidence that supports their claim that Gilead withheld a safer HIV drug.
Throughout Stribild’s time on the market, Gilead representatives had claimed that it was “risk-free” and posed no threat to kidney health or bone density.11 Not only was the U.S. Food and Drug Administration (FDA) reprimanding Gilead for these claims; Gilead had known of TDF’s risks through its research and development of TAF. This research and development had showed the absorption rates of the two drugs and the much higher risks of TDF. In fact, Gilead had begun emphasizing those risks as soon as it announced its discovery and subsequent release of TAF-based HIV drugs.12
Stribild lawsuits claim that Gilead caused victims to suffer dreadful injuries by withholding information on the risks of TDF.
4. What compensation can victims obtain?
2022 Stribild lawsuit plaintiffs are seeking compensation for the:
- Medical expenses they have already accumulated,
- Medical bills they are likely to accrue in the future as they recover,
- Wages they have lost during their recovery from Stribild’s side effects,
- Reduced earning capacity from the health effects of Stribild’s side effects,
- Physical pain and emotional and mental suffering of those side effects, and
- Loss of companionship suffered by the victim’s family and loved ones.
Many lawsuits also demand punitive damages to punish Gilead for its egregious conduct in withholding a safer drug from the market.13
5. Is there litigation against Gilead’s other HIV drugs?
Yes. In addition to Stribild, the pharmaceutical company Gilead Sciences, Inc. has also produced other HIV drugs that contain TDF and cause the same serious side effects. The TDF MDL involves all these medications, which are:
The HIV drugs that Gilead has released more recently have been based on TAF, rather than TDF. While TAF is likely a safer alternative, it can still cause major side effects. Gilead’s TAF-based line of HIV medication includes:
- Genvoya, the TAF-based pill that Gilead has urged Stribild users to switch to, and which has spawned its own set of Genvoya lawsuits,
- Vemlidy, and
- Willem D.F. Venter, June Fabian, and Charles Feldman, “An overview of tenofovir and renal disease for the HIV-treating clinician,” South African Journal of HIV Medicine 19(1):817 (2018).
- Pablo Rivas, Jorge Polo, Miguel de Górgolas, Manuel L Fernández Guerrero, “Drug points: Fatal lactic acidosis associated with tenofovir,” British Medical Journal 327:711 (2003).
- Iwen F Grigsby, Lan Pham, Louis M Mansky, Raj Gopalakrishnan, and Kim C Mansky, “Tenofovir-associated bone density loss,” Therapeutics and Clinical Risk Management 6:41-7 (2010).
- Scherzer R, Estrella M, Li Y, Choi AI, Deeks SG, Grunfeld C, Shlipak MG, “Association of tenofovir exposure with kidney disease risk in HIV infection,” AIDS 26(7):867-75 (April 24, 2012) and Steve Tokar, “Tenofovir, Leading HIV Medication, Linked with Risk of Kidney Damage,” University of California – San Francisco News Report (February 10, 2012).
- Martinez v. Gilead Sciences, Inc., No. BC 705063 (Cal. filed May 8, 2018).
- See e.g., Lujano v. Gilead Sciences, Inc., No. BC 702302 (Cal. filed May 18, 2018).
- FDA Approval Letter for TDF.
- See Martinez v. Gilead Sciences, Inc., No. BC 705063 (Cal. filed May 8, 2018) (complaint).
- Melody Petersen, “Patients sue Gilead, saying drug company intentionally delayed safer HIV medicine,” Los Angeles Times (May 9, 2018).
- See note 9.
- See note 8.
- See notes 8 and 9.
- Plaintiffs’ complaint at 17, Lujano v. Gilead Sciences, Inc., No. BC 702302 (Cal. filed May 18, 2018).

domain: medical


text:
(129f) Noncanonical Amino Acids for Temporally-Resolved Proteomic Profiling
AIChE Annual Meeting
Monday, November 17, 2014 - 2:06pm to 2:24pm
Noncanonical amino acids (ncAAs) can be incorporated into cellular proteins on a proteome-wide basis, allowing chemical modification of thousands of proteins simultaneously. Incorporation of ncAAs bearing bioorthogonal “clickable” functional groups in a well-defined pulse makes newly synthesized proteins easy to distinguish from old proteins (those produced pre-pulse). This approach allows selective analysis of the nascent proteome: labeled proteins can be conjugated to fluorescent probes for visualization, or to affinity tags for enrichment and identification by mass spectrometry. This method, termed bioorthogonal noncanonical amino acid tagging (BONCAT), is a powerful approach for measuring patterns of protein synthesis in response to biological cues.
We combine BONCAT with the quantitative proteomics technique pulsed stable isotope labeling with amino acids in cell culture (pSILAC), yielding a proteomic approach that can both enrich and quantify proteins synthesized during a ncAA pulse. The combined BONCAT-pSILAC approach provides accurate quantitative information about rates of cellular protein synthesis on time scales of minutes. We use this method to quantify 1400 HeLa cell proteins produced during a 30 minute pulse, a time scale inaccessible to isotope labeling techniques alone. Potential artifacts due to the incorporation of ncAAs can be reduced to insignificant levels by limiting the extent of amino acid replacement. We find no evidence for artifacts in protein identification in experiments that combine the BONCAT and pSILAC methods.


text:
$1,000 Deposit Will Benefit 3 out of 4 Children Born in 2021.
Proposal Mirrors Legislation Introduced at the Federal Level by U.S. Senator Cory Booker.
Governor Phil Murphy today joined with legislators, faith leaders, and community advocates to highlight the importance of the Baby Bonds initiative in his Revised Fiscal Year 2021 Budget, which will provide a $1,000 deposit into an account for every baby born into a household with income less than 500 percent of the Federal Poverty Level ($131,000 for a family of four) in 2021.
Advanced at the federal level by U.S. Senator Cory Booker and prominent economists, the proposal is a critical investment to address longstanding economic and race-based wealth disparities that have only been further exacerbated by the COVID-19 pandemic.
“While COVID-19 has had an enormous financial impact on many New Jersey families, it has laid bare systemic inequities that have disproportionately denied families of color an equal chance to achieve upward mobility,” said Governor Murphy. “To emerge from this crisis as a stronger, fairer, and more resilient state, we need to lay the foundation for the next generation of New Jerseyans to secure their spot in the middle class and break the cycle of economic inequality.”
“We are seeing wealth inequality at its highest point in decades, which is severely limiting equal opportunity for many Americans,” said U.S. Senator Cory Booker. “In a country as wealthy as ours, every child should have an opportunity to build assets, create wealth, and achieve upward mobility. As I continue working to address the racial wealth gap on the federal level, I applaud Governor Murphy for prioritizing economic inequality and introducing this bold proposal aimed at leveling the playing field for children in New Jersey growing up in low-income families.”
The parameters of the proposal require newborns to either be born in New Jersey or born to parents who are New Jersey residents to be eligible for an account. Baby Bond recipients would have to be New Jersey residents to withdraw funds from their accounts, although they would not have to spend these funds in New Jersey.
Under the Governor’s initiative, the State would establish the Baby Bonds Initiative Fund, which would include individual accounts for eligible residents. Funds in each child’s account would be invested so that they generate returns that are at least equivalent to thirty-year US Treasury bonds. Recipients would be able to withdraw funds when they turn 18 for allowable uses that build wealth such as homeownership, higher and continuing education, or entrepreneurship.
“This innovative proposal will help create a nest egg for every baby born in New Jersey whose family may struggle to start their child off on the right financial foot,” said Congressman Bill Pascrell. “As we work to eliminate systemic issues for building wealth and financial well-being exacerbated by this crisis, baby bonds look out for the next generation of New Jerseyans on the horizon. Now, more than ever, we need to empower the next generation to start on a level playing field and give them the tools to build a better world for themselves and their children. I applaud Governor Murphy for the courage, wisdom, and far-sighted vision to bring forward this proposal that will change lives.”
“With so many New Jerseyans in communities of color struggling to stay afloat right now, saving money for a child’s future is not an option,” said Senator Ronald C. Rice. “Baby bonds are simply an investment in the next generation of our state. Generational poverty holds back far too many people, and the Governor’s proposal is a creative way to support working class families at a time of great need.”
“Baby bonds are a critical investment in our children’s future,” said Senator Nellie Pou. “Now is the time to think big and to support working families across our state. This proposal will provide a financial foundation for children who need it most.”
“Even prior to the COVID-19 pandemic and the ensuing nationwide economic crisis, too many New Jersey families struggled with poverty and a lack of access to educational, social, and economic opportunities,” said Assemblyman Benjie Wimberly. “The need for investment in our communities and our children has never been greater. Baby bonds are a down payment on the success of our future generations and I support the Governor’s proposal.”
“Creating a healthy financial start for future generations is pivotal to the future of New Jersey. This means implementing wealth-building strategies such as saving for a college education to limit loan debt, encouraging homeownership, and ensuring greater access to capital,” said Assemblywoman Shavonda Sumter. “The glaring disproportionate access to all of the above has had a grave economic and generational impact on working poor families. The high-level investment of Baby Bonds in New Jersey’s children will make the state a leader in the nation in eradicating poverty and reducing persistent economic disparities.”
“One of the first initiatives I started working on was developing the framework for a Baby Bonds program in the Garden State,” said Assemblyman William Moen.“While COVID19 has affected all of our lives in different ways, it has made it ever more clear the need to narrow the income inequality gap and continue to help New Jersey families reach income equity. Now is the time to start this conversation. I commend the Governor on this proposal, and look forward to working alongside him and the legislative leadership to advance this issue.”
“It’s my pleasure to support this important initiative for baby bonds and to lookout for future generations with an understanding of the wealth disparity that has long haunted the Black community,” said Reva Foster, Chair of the New Jersey Black Issues Convention. “With this initiative, future generations will have a leg up and be able to secure a home, continue higher education, or start a business. All of this will help not only the recipients but there community will inspire others to do the same.”
“For working families across our state who live paycheck to paycheck, a baby bond will give their children an opportunity to break the cycle of income inequality,” said First Lady Tammy Murphy. “And, it will send every child in New Jersey the clear message that we believe in them, and they should believe in themselves too. This is our chance to make sure that every child on the brink of adulthood has both a dream and the start of a plan to pursue that dream.”

domain: sociology


Hawk Performance’s HP Plus brake pads were developed and manufactured especially for sport driving applications such as autocross, Solo II, and “track day” events. Superior Braking.
The HP Plus pads utilizes a unique Ferro-Carbon, high-tech friction material that has an extremely high friction coefficient making the pads "fade resistant" under repetitive, heavy braking conditions for excellent stopping power. Application.
This set of Hawk HP Plus Performance Brake Pads are specifically designed to be used on the front of the 2005 to 2014 V6, GT and Bullitt Mustangs. Does not fit Shelby GT500 or BOSS Mustangs. Set includes driver-side and passenger-side brake pads.
||Ultra Low Dust
Technical Note. The Hawk HP Plus Performance Brake Pads are not intended for everyday street/highway driving as they utilize a high-temperature specific compound for optimal performance under race braking conditions.
2005, 2006, 2007, 2008, 2009, 2010, 2011, 2012, 2013, 2014 Details >>


text:
Recruitment in Orissa
Orissa State Govt Jobs
How to Apply for Police Constable Jobs in Orissa Police for 2011
Orissa State Govt Jobs
: 10 (
In this 2011 year Orissa police is recruiting 1878 police constables in all the districts of Orissa. If you are confused about how to apply for Police Constable Jobs in Orissa Police for 2011 then read the below given article to get full details.
Application Procedure for Police Constable Jobs in Orissa Police for 2011
You have already seen the
Physical examination Centre for police constable recruitment in Orissa
. Now you can get the details about the procedure of applying for Police
Constable Jobs in Orissa Police for 2011
Interested applicants can get the application form from respective Superintendent of police office or Police commissionrate office. They can also Get the
from www.bpspaorissa.gov.in. No other types of forms will be accepted. All types of applicants have to submit Rs 25 for the application form in terms of Bank Draft/ Indian Postal Order. Applicants who have downloaded the application form from website also have to submit the amount. Like this all applicants (except SC/ST applicants) have to submit Rs 100 towards Examination fees in terms of Bank Draft or Indian postal Order.
All Bank drafts and Indian Postal Orders should be drawn in favor of "Chairman, State Selection Board, Odisha Police, Cuttack" and it should be payable at Cuttack. Applications received without IPO or Bank draft will not be taken into consideration and those will be rejected. All the applicants have to send the IPO/Bank draft and all copies of certificates through Post or they can submit these at their Superintendent of Police/ Police Commissioner office on or before 07.10.2011 cause 07 October 2011 is
last date for applying for Police Constable Jobs in Orissa Police for 2011
If you want to apply for this
recruitment by Orissa police
Application Form for Police Constable Job in Orissa Police for 2011
1878 Police Constable Vacancy in Orissa Police for 2011
Physical Test Examination Centres of Police Constable Recruitment in Orissa 2011
ASI Operator Recruitment in Orissa Police Signals by OSSC for 2011
Qualification and Documents Needed for Police Constable Recruitment in Orissa Police for 2011
1621 Soldiers Vacancy in Orissa Special Armed Police OSAP for 2011
1650 Constables Job Vacancies in India Reserve Battalion IRB for Orissa
Army Recruitment Rally By Army Recruiting Office Gopalpur in Orissa
CRPF Constable Recruitment in Odisha for 2011
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domain: law


text:
Image: Paula Dawson using Holoshop haptic interface to generate content for digital holographic prints.
Holoshop combines innovative technologies and processes for creating real-time 3D digital content for holograms and other 3D display systems, with virtual tools enabling the direct hand-drawing of subjects, mark by mark. The Holoshop project creates the foundations for a new method of 3D content creation that factors the key perceptual differences between the level and type of detail required of a two-dimensional or three-dimensional media into a process of 3D line drawing construction. Using a haptic (tactile) interface, Holoshop software registers the users’ gestures as a digital 3D scene. This software enables three-dimensional templates made in other softwares to be imported and used as guides for drawing. It also enables aesthetic choices such as colour and line, and can be modified to track gestural inputs such as speed.
In this forum, chaired by ISEA2013 Keynote Professor Brian Rogers, investigators and researchers of the Holoshop project will discuss the development and perceptual effects of Holoshop. Detailing the development of the project across varied platforms, the speakers will compare the unique perceptual effects created by Holoshop software across holographic, three-dimensional, stereoscopic and virtual mediums.
Keynote speaker and chair: Brian Rogers
Speakers: Paula Dawson, Joanna Szymanska, Fay Hudson, Hiroshi Yoshikawa, John Stavrakakis, Masa Takatsuka, Philip Lange, Karen Kriss
Professor Brian Rogers teaches in the Department of Experimental Psychology at the University of Oxford. He is acknowledged as one of the foremost experts on 3D perception. In his research he has shown that motion parallax can be as effective as binocular disparities as a source of 3D information, and recently he has been studying how the many different sources of 3D information interact and combine to create the perception of 3D structure and layout. Within Holoshop, he advises on the effectiveness of research progress across a range of 3D platforms’ display systems – stereo, structure-from-motion, CGH and holo-video – assessing their advantages and disadvantages. He is also involved in the design of data collection and collation from visitor participation with the Holoshop: Drawing and Perceiving in Depthexhibition and workshops for ISEA2013.
Associate Professor Paula Dawson teaches at the College of Fine Arts, UNSW. She has pioneered large volume optical hologram recording and the investigation of spatial and temporal effects of pictorial agents in optical, digital and computer generated hologram imagery. Paula leads the project, ensuring that the group’s activities are wholly integrated, designing the virtual drawing tools, and undertaking experiments in drawing sequences through the practical use of the Holoshop software and the haptic interface, the Phantom. Paula creates the scene files and supervises their testing across computer generated hologram (CGH), digital hologram, holo-video, stereo and related 3D display systems.
Associate Professor Masa Takatsuka teaches at VisLAB, School of IT, University of Sydney. He has pioneered the development of neural networks for encoding individual characteristics of objects, including free-form surfaces, which are particularly relevant to this project. He has also developed a texture-based 3D shape encoding technique that utilises the geometric shading technology available in recent Graphical Processing Units, and oversees the design and development of computational algorithms and software design which enable Holoshop haptic interface to be used for virtual drawing purposes.
Professor Hiroshi Yoshikawa teaches at Nihon University, Japan. He is an expert in high-speed calculation on computer-generated holograms. With pixel number exceeding 100 billion for a 150 mm x 150 mm hologram, his skills in fast calculation are crucial to the generation of such huge holograms for the Holoshop project. He also adapts the mathematical generation of the holographic fringe pattern of virtual drawings to match the parameters for electrobeam lithography at the Australian National Fabrication Facility, University of New South Wales.
Dr Joanna Szymanska is a highly skilled nano-fabrication engineer at the Australian National Fabrication Facility, UNSW. She has developed a process of creating computer generated holograms (CGH) using electron beam lithography, and has successfully written several CGHs. One of them, 3 x 3mm in size, will be shown in the Holoshop: Drawing and Perceiving in Depth exhibition for ISEA2013.
Dr John Stavrakakis is a specialist in 3D computer graphics and networking at VisLAB School of IT, University of Sydney, and holds a PhD in Computer Science from the University of Sydney. John has broad technical skills used in projects that pursue the educational, research and industrial application of technologies. He is currently developing the Holoshop software supporting the use of haptic devices and visual feedback.
Phillip Lange, Visual Effects Compositor is a Visual Effects Compositor based in Sydney with over a decade of experience working on feature films and television projects. He has worked both locally and internationally and most recently on Great Gatsby, Harry Potter and the Deathly Hollows, and Happy Feet 2. Karen Kriss, Lecturer College of Fine Arts, UNSW teaches at the College of Fine Arts,UNSW and has as worked as a Visual Effects producer and Animator on feature films and short animation productions in Sydney, Montreal, Toronto and Los Angeles including award winning films Happy Feet and Lust Caution. She is undertaking research with a Master of Fine Arts at the College of Fine Arts, UNSW.
Karen Kriss teaches at the College of Fine Arts, UNSW and has as worked as a Visual Effects producer and Animator on feature films and short animation productions in Sydney, Montreal, Toronto and Los Angeles, including award winning films Happy Feet and Lust Caution. She is currently undertaking research for a Master of Fine Arts at the College of Fine Arts, UNSW.

domain: design


text:
On Friday January 15, 2010, the Third Circuit Court of Appeals (the federal appeals court for New Jersey, Pennsylvania and Delaware) heard oral argument in Miller v. Skumanick, a case involving “sexting.” This practice, which has become increasingly popular among teenagers in recent years, refers to the sending or posting sexually suggestive text messages and images — including nude or semi-nude photographs — via cell phone or over the Internet.
The case arose when Pennsylvania school officials discovered that several students had exchanged semi-nude images of themselves via cell phone. School officials turned over the cell phones containing the images to law enforcement authorities, and the District Attorney threatened to prosecute the teenage girls in the photographs on charges of child pornography unless they took a class he had designed to address the dangers of “sexting.” Through their parents, the students filed suit against the District Attorney, claiming that his actions had infringed on their First Amendment rights.
Earlier last year, a federal judge granted a temporary restraining order that prevented the District Attorney from initiating criminal charges against the students. The full text of the lower court’s opinion is available by clicking here. The Third Circuit will consider whether the lower court’s order should be upheld, which will require the court to determine whether the offending images can constitute child pornography under the law. To read more about this case, please click here.


DIY Steel Pipe Shelf
It gives the impression that the shelf has been installed around the plumbing.
When mounting the shelf to a plasterboard wall, find the studs and mark them on the wall using masking tape, then use the distance between the studs to determine the length of the lower pipe.
DIY TIP: Mount the shelf to the studs or use hollow wall anchors through plasterboard.
Mark two pipe holes on the shelf with the centres spaced the same distance apart as the stud centres, and 40mm from the front edge.
Drill the holes using a 15mm spade bit.
Make a third hole in line with the first hole on the right, 90mm from the front edge.


text: Sweet Angeline I turned my little sun room into a studio and that was where these images were taken. I absolutely love natural light and love the feel of these images. Sweet Angelina was a joy to photograph!! Show Hide 8 comments Add a comment... Your email is never<\/em> published or shared. Required fields are marked * Post Comment Lisaaawwww – she’s beautiful!!! JohanneI agree ;o) Michael NearingJohanne, you are *SO* talented. Again, Thank-you so much! JohanneOh it was my pleasure 😉 It was great seeing you guys and the lovely Angeline – what a sweetheart! StephanieBeautiful baby & beautiful photos! Great work again!! SarahThere are those who give with joy, and that joy is their reward. – Kahlil Gibran Johanne your pictures are so beautiful you bring beauty and happiness to everyone, you are such an inspiration!! Nathalie BoudreauReally nice pictures!!! She’s sooo cute with her little polka dot dress!!! Olaoooooohhhhhhh…. she is sooooooo gorgeous – FANTASTIC photos!!!


United States – 01-01-2019 (PRDistribution.com) — Move Honcho, an online moving resource company, has launched a new website in the Salt Lake City market on New Year’s Day. MoversSLC.com offers moving estimates from certified local movers and do-it-yourself moving tips catered to SLC residents.
Move Honcho was founded by a former part-time college mover with a background in design and writing. He noticed that many people were lost when it came to moving – both with how to move themselves and with how to hire the right moving company. From that observation sparked Move Honcho. From quotes from Salt Lake City moving companies to advice on how much to tip movers or how to pack a box, the sites mission is to simplify the moving process by educating people and streamlining their mover search process. All movers in their network are screened to meet the proper licensing and insurance requirements, as well as have a consistent track record of satisfied clients. While there are similar services out there, Move Honcho has the advantage of being founded by a former mover and catering heavily to specific geographic markets.
For the original news story, please visit https://prdistribution.com/news/moving-resource-company-move-honcho-launches-new-website-in-salt-lake-city.html.


text:
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