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Supplementary Figure S6 from Caspase-8 Promotes Cell Motility and Calpain Activity under Nonapoptotic Conditions

Caspase-8 is a proapoptotic protease that suppresses neuroblastoma metastasis by inducing programmed cell death. Paradoxically, caspase-8 can also promote cell migration among nonapoptotic cells; here, we show that caspase-8 can promote metastasis when apoptosis is compromised. Migration is enhanced by caspase-8 recruitment to the cellular migration machinery following integrin ligation. Caspase-8 catalytic activity is not required for caspase-8–enhanced cell migration; rather, caspase-8 interacts with a multiprotein complex that can include focal adhesion kinase and calpain 2 (CPN2), enhancing cleavage of focal adhesion substrates and cell migration. Caspase-8 association with CPN2/calpastatin disrupts calpastatin-mediated inhibition of CPN2. In vivo, knockdown of either caspase-8 or CPN2 disrupts metastasis among apoptosis-resistant tumors. This unexpected molecular collaboration provides an explanation for the continued or elevated expression of caspase-8 observed in many tumors. [Cancer Res 2009;69(9):3755–63]

Significant caspase-8 activity has been found in normal and certain tumor cells, suggesting that caspase-8 possesses an alternative, nonapoptotic function that may contribute to tumor progression. In this article, we report that caspase-8 promotes cell motility. In particular, caspase-8 is required for the optimal activation of calpains, Rac, and lamellipodial assembly. This represents a novel nonapoptotic function of caspase-8 acting at the intersection of the caspase-8 and calpain proteolytic pathways to coordinate cell death versus cell motility signaling. (Cancer Res 2006; 66(8): 4273-8)

XLSX file - 146K, Table S1- Data on the genes that differ in expression between primary tumors Th- MYCN, Caspase-8 wild type levels with no BMM, and primary tumors Th-MYCN, deficient caspase-8 with BMM (p

PDF file - 709K, Figure S1. Correlation of relative mean fold changes in expression of genes and microRNAs of the mouse tumor groups by expression- array and by quantitative RT-PCR. Figure S2. Levels of caspase-8 protein in primary tumors and BM secondary tumors. Figure S3. Primary tumors from wild-type or caspase-8 deficient (fl/ko, fl/fl or wt/ko caspase-8) mice had neuroblastoma morphology and stained positive for neuronal markers typical for neuroblastoma. Figure S4. Age at maximum tumor burden of neuroblastoma primary tumors in the Th-MYCN mouse groups. Figure S5. Measuring apoptosis levels by immunostaining for cleaved caspase-3. Figure S6. MYCN expression in the primary Th-MYCN, Th-Cre neuroblastoma tumors. Figure S7. Secondary tumors observed in both the Th-MYCN Caspase-8 wt/wt and the Th-MYCN deficient caspase-8 mouse models.

XLSX file - 77K, Table S3- Data on the microRNAs that differ in expression between primary tumors Th- MYCN, Caspase-8 wild type levels with no BMM, and primary tumors Th-MYCN, deficient caspase-8 with BMM (p

Supplementary Figures 1-5 from Caspase-8 Association with the Focal Adhesion Complex Promotes Tumor Cell Migration and Metastasis

Supplementary Figure Legends S1-S10 from Caspase-8 Promotes Cell Motility and Calpain Activity under Nonapoptotic Conditions

Neuroblastoma, the most common extracranial pediatric solid tumor, is responsible for 15% of all childhood cancer deaths. Patients frequently present at diagnosis with metastatic disease, particularly to the bone marrow. Advances in therapy and understanding of the metastatic process have been limited due, in part, to the lack of animal models harboring bone marrow disease. The widely used transgenic model, the Th-MYCN mouse, exhibits limited metastasis to this site. Here, we establish the first genetic immunocompetent mouse model for metastatic neuroblastoma with enhanced secondary tumors in the bone marrow. This model recapitulates 2 frequent alterations in metastatic neuroblasoma, overexpression of MYCN and loss of caspase-8 expression. Mouse caspase-8 gene was deleted in neural crest lineage cells by crossing a Th-Cre transgenic mouse with a caspase-8 conditional knockout mouse. This mouse was then crossed with the neuroblastoma prone Th-MYCN mouse. Although overexpression of MYCN by itself rarely caused bone marrow metastasis, combining MYCN overexpression and caspase-8 deletion significantly enhanced bone marrow metastasis (37% incidence). Microarray expression studies of the primary tumors mRNAs and microRNAs revealed extracellular matrix structural changes, increased expression of genes involved in epithelial to mesenchymal transition, inflammation, and downregulation of miR-7a and miR-29b. These molecular changes have been shown to be associated with tumor progression and activation of the cytokine TGF-β pathway in various tumor models. Cytokine TGF-β can preferentially promote single cell motility and blood-borne metastasis and therefore activation of this pathway may explain the enhanced bone marrow metastasis observed in this animal model. Cancer Res; 73(13); 4086–97. ©2013 AACR.

XLSX file - 175K, Table S2- Data on the genes that differ in expression between primary tumors Th- MYCN, Caspase-8 wild type with no BMM, and primary tumors Th-MYCN, deficient caspase-8 without BMM (p

The effect of 20 MeV electron radiation on the lifetime of the silicon carbide power MOSFETs was investigated. Accelerated constant voltage stress (CVS) was applied on the pristine and irradiated devices and time-to-breakdown ( T BD ) and charge-to-breakdown ( Q BD ) of gate oxide were extracted and compared. The effect of electron radiation on the device lifetime reduction can be observed at lower stress gate-to-source voltage ( V GS ) levels. The models of T BD and Q BD dependence on the initial gate current ( I G0 ) are proposed which can be used to describe the device breakdown behaviour. peerReviewed

The title diradical was synthesized and investigated by ESR and UV-vis spectroscopy. It was found to have a lifetime of weeks even in the presence of oxygen, and even survives brief heating in toluene up to about 60 °C. In the UV-vis spectrum, the diradical showed reversible thermochromic behavior in the -10 to 50 °C range. In the ESR spectrum, hyperfine analysis showed nearly isolated behavior by the phenoxyl and nitroxide spin carrying units. Upon warming, additional, broad lines appeared at the expense of the lower temperature lines. This temperature behavior was reversible over the 0 to 50 °C range, so long as the raised temperatures were not maintained for long time periods. The spectral behavior is interpreted as being due to temperature-dependent conformational effects on the exchange coupling between the spin carrying units, i.e.

A series of donor-acceptor (D–A) type conjugated molecules was synthesized, with structures D–π–A or A–π–D–π–A where A = N-hexylpyridinium-4-yl and D = electron donor type units, with A and D linked by π-conjugating units: the D–π–A systems are design-testing models for optimizing the A–π–D-π-A systems, where the latter are conjugatively extended variants of viologen. UV–vis absorption spectra and molar absorptivities were determined in varying solvents, and spectra for neat films. The donor units D comprised phenylene, 2,5-thienylene, 3,4-ethylenedioxythiophen-2,5-diyl (EDOT), 1-N-methyl-2,5-pyrrolediyl, and CH CH units. The system with strongest absorptivity at a relatively long absorption maximum and onset was the extended viologen 4-(1-hexyl)pyridinium-CH CH-(2,5-EDOT-CH CH-)-(2,5-thienylene-CH CH )-(2,5-EDOT-CH = CH-)-4-(1-hexyl)pyridinium dibromide, with λmax(methanol) = 576 nm (278,000 M−1 cm−1), low energy onset at 683 nm (1.82 eV). Photoluminescence quantum yields in methanol mostly were quite weak, with solvatochromic behavior indicating strong intramolecular charge transfer character.

X-ray diffraction results for single crystals of N,N-diarylanilinosquaraines (SQ-TAAs) 2,4-bis(4-bis(p-R-phenyl)amino)-2,6-dihydroxyphenyl)cyclobuta-1,3-diene-1,3-bis(olate) for R = F (SQ-TAA-F) and R = n-hexyl (SQ-TAA-C6) are reported. SQ-TAA-F (an ambipolar charge transporter) forms π-stacked dyads intercolated by cross-stacked molecules, a unique motif among the systems studied. Its [C]–H···F contacts induce this intermolecular packing and may account for its ambipolar charge transport behavior. SQ-TAA-C6 (a hole charge transporter) forms strongly slip-stacked 1-D chains aligned along molecular long axes. SQ-TAA-C6 exhibits packing of its conjugated core structure that is similar to those for SQ-TAAs with R = H and R = OH, which also exhibit qualitative hole transport properties. SQ-TAA-C6 has one n-hexyl group that is not fully extended, but all of its alkyl groups pack into alkyl rich planes that alternate with SQ-TAA core rich planes to form the overall lattice.

Thermoelectric (TE) measurements were compared for iodine-doped composites of carbon nanotubes (CNTs) with poly(3-hexylthiophene-2,5-diyl) (P3HT) having different degrees of regioregularity. Regioregular P3HT gave much higher power factors, showing the need of regioregularity in the polymer for best TE results. TE power factors (PFs) up to 148 μW m−1 K−2 were obtained by optimizing multiple factors, including: (i) P3HT sample type, (ii) relative amount and type of CNT, (iii) sample fabrication solvent, and (iv) composite doping duration. The TE performance improvements in the composites arose from greatly increased electrical conductivity by doping and CNT addition, coupled with limited decreases in Seebeck thermopower. Extensive network formation was seen by scanning electron microscopy (SEM) in composites with higher CNT fraction, which is believed to provide better charge transport pathways to improve PFs. Multiwall CNT composites showed much lower PFs compared to singlewall CNT composites, attributable (at least in part) to poor nanotube dispersion.

The synthesis, device fabrication, and electric poling of novel amino-functionalized fulleropyrrolidines in polymer solar cells is reported. Systematically varying the tether lengths between the fullerene cage and tertiary amines provides insight into enhanced device efficiency and stability after poling. DFT calculations in an external electric field result in alignment of molecular dipole moments, which corroborates our hypothesis that electrotropic polarization causes an increased built-in electrostatic potential difference across the device, and results in enhance performance.

The geometry and electronic structures of four cyclopentadithiophene (CPD) derivatives in their crystals are systematically studied through density functional theory (DFT) calculations. The factors influencing the CPDs' semiconductor nature and charge carrier mobility are analyzed in detail, revealing the different roles of fluorine and dicyanomethylene in the CPDs examined in this study. Malononitrile substitutions show a large effect in lowering the molecular LUMO energy level but induce a smaller charge transfer integral due to the resulting reduced crystal order caused by the resulting steric hindrance. In comparison, although F substitution lowers the LUMO level more weakly than C(CN)2 substitution, it allows the molecule to retain a more planar configuration with more orbital delocalization, and the crystals of the F substituted compounds are more ordered than those of the C(CN)2 substituted compounds. Overall, this comparison can explain why F substitution is a more effective method to cause a transition of a p-type semiconductor to n-type. Crystal ordering plays a significant role in determining the charge transfer mobility in devices. In spite of the advantage offered by dicyanomethylene substitution in terms of charge injection barrier compared to fluorine substitution, the reduced crystal order caused by C(CN)2 limits the charge transfer in CPDs with C(CN)2 substitution. The validity of these findings can be generalized, which is expected to contribute to the rational design of n-type semiconductors based on CPDs.

Autoimmune diabetes (T1D) is caused by the immune-mediated destruction of insulin-producing beta cells in the pancreas leading to hyperglycemia. Apoptosis is the main mechanism responsible for beta cell demise, including Fas-receptor engagement and Perforin release as essential death executers. However, the chronic inflammatory milieu pervading pancreatic islets prior to diabetes onset, alters beta cells function and primes beta cells for apoptosis long before the debut of the disease. To identify those genes that are targeted by inflammation and are causally related to beta cell fitness and viability we have used the microarray technology. We have identified several candidate genes being downregulated by inflammation, one of this genes is Cdk11, a cyclin-dependent kinase which is involved in transcription (CDK11p130), mitosis and apoptosis (CDK11p58). L-type cyclins are the partners for Cdk11p130, and Cyclin D3 is the partner of CDK11p58. The Cdk11 gene, when is transcribed, generates a single mRNA form, that can be translated either into CDK11p130 or CDK11p58 by a differential IRES (Internal Ribosome Entry Site) usage, being CDK11p58 only expressed during mitosis, while CDK11p130is expressed during all cell cycle phases. We have addressed the role of CDK11 in autoimmune diabetes using the T1D-prone NOD mouse model hemideficient in CDK11, since the CDK11 null mutation is embryonically lethal. First of all, we have confirmed the effect of inflammation on Cdk11 mRNA expression in islet cells by submitting NODSCID mice to adoptive transfer of increasing amounts of diabetogenic NOD splenocytes, and, observed that Cdk11 expression in islet endocrine cells is downregulated by inflammation in a dose-response dependent manner.

Lipid oxidation is one of the major causes of oil deterioration causing off-flavors and consumer rejection. Fast, easy, and dependable assays for predicting lipid oxidation rates in foods are important for shelf-life prediction. In this study, an electron paramagnetic resonance (EPR) spin-trapping technique with N-tert-butyl-α-phenylnitrone (PBN) was tested to determine the lag phase of lipid oxidation in stripped soybean oil (SSO), SSO with added α-tocopherol, and commercial soybean, canola and corn oils. EPR intensity of spin-trapped products from SSO correlated well with lipid hydroperoxides formation for samples stored at 37 and 55 °C respectively. When the antioxidant α-tocopherol was added, the EPR signal intensity of oil samples increased—indicating sample deterioration—after 50–65% of α-tocopherol was consumed. When using the EPR method with commercial soybean, canola or corn oil stored at 55 °C, there was a poor relationship between EPR intensity and lipid hydroperoxides lag phases. However, a linear correlation was found between EPR signal intensity and hexanal formation. For example, EPR signal intensity lag phases were 5, 13 and 27 days for soybean, canola and corn oils, respectively which was similar to the hexanal lag phases of 5, 13 and 25 days for the same oils. The EPR spin-trapping assay method has several advantages over headspace hexanal measurements, especially with regard to easier sample handling and shorter analysis times.

The first syntheses of bis(nitronyl nitroxide) and bis(iminoyl nitroxide) (diNN, diIN) biradicals linked through rigid acene core conjugating anthracene (A) and anthraquinone (AQ) units are reported. Computational modeling predicts weak intramolecular exchange in AQ-linked systems, but A-linked biradicals to have ground state multiplicities consistent with the Borden-Davidson disjointness model. Solution electron spin resonance spectra showed inter-radical exchange-coupled triplet states, except for 2,6-AQ biradicals showing isolated spin spectra. Crystallography of the A-linked biradicals shows a key role for inter-radical contacts for molecular packing. DiINs showed lower-dimensional dyad packing with disorder at the radical units: the conformationally more symmetrical diNNs gave staircase one-dimensional or brickwork two-dimensional lattices. Core anthracene unit stacking was only seen in two systems with bromine on the central anthracene ring: the (large) bromine occupies alternate side placement in d...

Binary polymer nanoparticle glasses provide opportunities to realize the facile assembly of disparate components, with control over nanoscale and mesoscale domains, for the development of functional materials. This work demonstrates that tunable electrical percolation can be achieved through semiconducting/insulating polymer nanoparticle glasses by varying the relative percentages of equal-sized nanoparticle constituents of the binary assembly. Using time-of-flight charge carrier mobility measurements and conducting atomic force microscopy, we show that these systems exhibit power law scaling percolation behavior with percolation thresholds of ∼24-30%. We develop a simple resistor network model, which can reproduce the experimental data, and can be used to predict percolation trends in binary polymer nanoparticle glasses. Finally, we analyze the cluster statistics of simulated binary nanoparticle glasses, and characterize them according to their predominant local motifs as (p(i), p(1-i))-connected networks that can be used as a supramolecular toolbox for rational material design based on polymer nanoparticles.

We present a combination of heat capacity and ac susceptibility measurements as a function of temperature and field, of a single crystal of the radical 2-(4,5,6,7-tetrafluorobenzimidazol-2-yl)-4,4,5,5-tetramethyl-4,5-dihydro-1H-imidazole-3-oxide-1-oxyl. Our results confirm a zero-field Neel temperature at T N (0) = 0.73 K and allow the determination of the critical field parameters H c (||) = 170 Oe and H c (⊥) = 306 Oe. Assuming the previously determined 1-D intrachain ferromagnetic exchange interaction of J intra /k = +22 K between radicals, the interchain antiferromagnetic interaction is calculated as z J inter /k ∼ −0.03 K.

Eight cyclopentadithiophene (CPD) based semiconductors are designed by moving phenyl/fluorine-phenyl groups from α to β positions of thiophenes of CPD. The semiconductor performances of these CPD derivatives with Cs and C2 symmetry (termed as CPD’ and CPD’’, respectively) and their unsubstituted CPD backbones are analyzed systematically through DFT calculations. Some CPD’/CPD’’ compounds show better charge transfer mobility than CPD. We analyze in detail the factors influencing the semiconductor nature of these CPD derivatives and their charge carrier mobilities, revealing the different roles of fluorine and dicyanomethylene and of the α and β position of phenyl on the thiophene rings. Our findings can be generalized to broader classes of organic semiconductors and are expected to contribute to the rational design of n-type semiconductors based on CPDs.

To determine and compare patients' willingness-to-pay (WTP) for tooth extraction and filling services in Tanzania and to assess the socio-demographic factors that are associated with such valuations.Contingent valuation survey utilizing an open-ended willingness-to-pay format was administered among 1522 outpatients in four regional hospitals in Tanzania. WTP for extraction and tooth filling services for various tooth categories were determined and compared using Mann-Whitney and Kruskal-Wallis tests. The association of WTP values with socio-demographic background factors was assessed using multiple regression analysis.The mean WTP amounts for tooth filling were Tanzania shillings (Tshs) 7,398 (3.4 US$) and Tshs 7,726 (3.5 US$) for anterior and posterior teeth respectively. The mean WTP for tooth filling services was lower than the average charged fees in dental facilities. The mean WTP amounts for tooth extraction were Tshs 5,448 (2.5 US$) and Tshs 6,188 (2.8 US$) for anterior and posterior teeth respectively. WTP amounts were shown to vary by age, income, outpatient status and previous experience with the dental services. Belonging in youngest age group (18-24 years) and having a high-income level was associated with increased odds for high WTP valuations irrespective of tooth and treatment types.WTP reveals a preference for tooth filling rather than extraction services in this population. More studies are needed to address the discrepancy between the stated preferences and utilization patterns for dental services.

The source of unoccupied Ti 3d states in the case of stoichiometric anatase structured (TiO2)n clusters has been investigated using ab initio methods. These unoccupied gap states appear for example in the case of a stoichiometric (TiO2)38 cluster. We show that the origin of these gap states is related to effective subcluster formation which gives rise to empty defect-like gap states, when these states are split off from conduction band. © 2015 Wiley Periodicals, Inc.

Recent epidemiologic papers are presenting prevalence data suggesting breaks and decreases in obesity rates. However, before concluding that the obesity epidemic is not increasing anymore, the validity of the presented data should be discussed more thoroughly. We had a closer look into the literature presented in recent reviews to address the major potential biases and distortions, and to develop insights about how to interpret the presented suggestions for a potential break in the obesity epidemic. Decreasing participation rates, the use of reported rather than measured data and small sample sizes, or lack of representativeness, did not seem to explain presented breaks in the obesity epidemic. Further, available evidence does not suggest that stabilization of obesity rates is seen in higher socioeconomic groups only, or that urbanization could explain a potential break in the obesity epidemic. However, follow-ups of short duration may, in part, explain the apparent break or decrease in the obesity epidemic. On the other hand, a single focus on body mass index (BMI) ≥ 25 or ≥ 30 kgm-2 is likely to mask a real increase in the obesity epidemic. And, in both children and adults, trends in waist circumferences were generally suggesting an increase, and were stronger than those reported for trends in BMI. Studies concluding that there is a recent break in the obesity epidemic need to be interpreted with caution. Reported studies presenting a break were mostly of short duration. Further, focusing on trends in waist circumference rather than BMI leads to a less optimistic conclusion: the public health problem of obesity is still increasing.

We study the interaction between dibromobianthryl (DBBA) and the Ag(100) surface using scanning tunneling microscopy and density functional theory. DBBA is prochiral on adsorption and forms racemic domains with molecular rows aligned with the substrate nearest-neighbor [011] and [01¯1] directions. Deposition at elevated temperatures leads to the formation of disordered meandering graphene nanowires of constant width.

Free-standing iodine-doped composite samples of poly[2-methoxy-5-(2-ethylhexyloxy)-1,4-phenylenevinylene] (MEH-PPV) with carbon nanotubes (NTs) showed thermoelectric (TE) power factors (PFs) up to 33 μW·m–1·K–2 after optimizing multiple factors, including: (1) sample fabrication solvent, (2) doping time, (3) average MEH-PPV molecular weight, (4) NT fraction in the composite, and (5) use of single-wall versus multi-wall nanotubes (SWNT and MWNT, respectively). Composite fabrication from halogenated solvents gave the best TE performance after iodine doping times of 2–4 h; performance drops substantially in ∼20 h doped samples. TE performance dropped after at least 24 h of removal from iodine vapor but was fully restored upon re-exposure to the dopant. Longer-chain MEH-PPV gave not only mechanically stronger films but also higher PFs in doped SWNT composites. MWNT composites gave low PFs, attributed to poor NT dispersion. Scanning electron microscopy showed increasingly extensive network formation as NT frac...

A single hydrogen bonding molecular assembly unit (benzimidazole = BIm) was used for solid state assembly of different types of radicals connected at different sites on the assembly unit. The effects of changing radical spin unit and molecular connectivity were tested by comparing tetramethylnitronylnitroxide (NN), dimethyloxoverdazyl (mV), and diisopropyloxoverdazyl (iV) spin units attached to BIm at either the 2-position or 5/6-position. The mV systems were not isolated with sufficient purity and stability for crystallographic or magnetic analysis, but chain-type hydrogen bonding was found in the crystal structures of all of the other, stable compounds. Close contacts are induced between radical groups in some cases by the hydrogen bonded chains, giving exchange behavior ranging from essentially isolated spin paramagnetism (BIm2iV) to antiferromagnetic interactions of (−)1–3 K.

The chemisorption of tellurium on atomically clean Cu(111) surface has been studied under ultra-high vacuum conditions. At room temperature, the initial stage of growth was an ordered 2 3 × 2 3 R30° phase (0.08 ML). An ordered 3 × 3 R30° phase is formed at 0.33 ML coverage of Te. The adsorption sites of the Te atoms on the Cu(111) surface at 0.08 ML and 0.33 ML coverages are explored by quantitative low energy electron diffraction (LEED) and density functional theory (DFT). Our results indicate that substitutional surface alloy formation starts at very low coverages.

Silicon wafers both without and with silicon(IV) oxide surface coverage were covered with benzene solutions of stable organic radical 3-(N-tert-butyl-N-aminoxyl)benzoic acid (mNBA). X-ray photoelectron spectroscopy supported the presence of the radical on both surface-cleaned (oxide-reduced) and oxide-covered surfaces. Optical waveguide spectroscopy showed that the radical retained its structure while adsorbed to the surface of the wafers, without noticeable decomposition. AFM and MFM imaging showed that the radical formed blocky particles with a change in rms roughness from 0.3 nm premodification to 1.7 nm postmodification on the surface-cleaned silicon. Similar experiments using oxide-coated silicon showed that the radical adsorbed to form much smoother layers, with a small change in rms roughness from 0.2 to 0.3 nm. Contact angle measurements of water on the premodified and postmodified samples showed a large, hydrophobic change in the silicon oxide surface but only a modest change in the surface-clean...