Mitochondria Dysfunction in Lung Cancer-Induced Muscle Wasting in C2C12 Myotubes

Aims: Cancer cachexia is a syndrome which results in severe loss of muscle mass and marked fatigue. Conditioned media from cachexia-inducing cancer cells triggers metabolic dysfunction in skeletal muscle, including decreased mitochondrial respiration, which may contribute to fatigue. We hypothesized that Lewis lung carcinoma conditioned medium (LCM) would impair the mitochondrial electron transport chain (ETC) and increase production of reactive oxygen species, ultimately leading to decreased mitochondrial respiration. We incubated C2C12 myotubes with LCM for 30 min, 2, 4, 24 or 48 h. We measured protein content by western blot; oxidant production by 2′,7′-dichlorofluorescin diacetate (DCF), 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF), and MitoSox; cytochrome c oxidase activity by oxidation of cytochrome c substrate; and oxygen consumption rate (OCR) of intact myotubes by Seahorse XF Analyzer. Results: LCM treatment for 2 or 24 h decreased basal OCR and ATP-related OCR, but did not alter the content of mitochondrial complexes I, III, IV and V. LCM treatment caused a transient rise in reactive oxygen species (ROS). In particular, mitochondrial superoxide (MitoSOX) was elevated at 2 h. 4-Hydroxynonenal, a marker of oxidative stress, was elevated in both cytosolic and mitochondrial fractions of cell lysates after LCM treatment. Conclusion: These data show that lung cancer-conditioned media alters electron flow in the ETC and increases mitochondrial ROS production, both of which may ultimately impair aerobic metabolism and decrease muscle endurance

Characterization of Defects and Designed Flaws in Metal Additive Manufacturing (AM) Parts with X-ray Computed Tomography (XCT)

Metal Additive Manufacturing (AM) has great potential to revolutionize manufacturing industries, but a reliable method to detect defects in AM-produced parts with complex internal structures must be developed prior to its widespread adoption. In this research, defects occurring in additively manufactured metal parts are characterized with X-ray Computed Tomography (XCT). In addition, the concept of metrological XCT is introduced, with the goal of providing more accurate dimensional measurements of the internal features. Preliminary experiments have been conducted toward the larger goal of evaluating XCT as a viable option for nondestructive evaluation of AM-produced components. Critical to this evaluation is the principle of the Probability of Detection (PoD), which has not been previously determined for typical AM defects using XCT. The first step in determining the PoD for XCT is to develop a suitable artifact with embedded features that are representative of the defects occurring in AM-produced parts. Two sets of samples were built by AM and their embedded defects measured by XCT. In the first set of samples, the chosen AM processing parameters were suboptimal, ensuring that defects would be present in the parts. These parameters were also varied to understand their effect on the resulting microstructure and defect formation. Measurements of porosity and the pore size distribution were determined from the XCT images. In the second set of samples, synthetic internal features were added, some approximating typical AM defects. Dimensional XCT was used to evaluate the quality of these features. Based on the results, a future experiment with the goal of estimating the PoD of critical AM defects with an XCT system is suggested

Proteomic Analysis of Media from Lung Cancer Cells Reveals Role of 14-3-3 Proteins in Cachexia

Aims: At the time of diagnosis, 60% of lung cancer patients present with cachexia, a severe wasting syndrome that increases morbidity and mortality. Tumors secrete multiple factors that contribute to cachectic muscle wasting, and not all of these factors have been identified. We used Orbitrap electrospray ionization mass spectrometry to identify novel cachexia-inducing candidates in media conditioned with Lewis lung carcinoma cells (LCM). Results: One-hundred and 58 proteins were confirmed in three biological replicates. Thirty-three were identified as secreted proteins, including 14-3-3 proteins, which are highly conserved adaptor proteins known to have over 200 binding partners. We confirmed the presence of extracellular 14-3-3 proteins in LCM via western blot and discovered that LCM contained less 14-3-3 content than media conditioned with C2C12 myotubes. Using a neutralizing antibody, we depleted extracellular 14-3-3 proteins in myotube culture medium, which resulted in diminished myosin content. We identified the proposed receptor for 14-3-3 proteins, CD13, in differentiated C2C12 myotubes and found that inhibiting CD13 via Bestatin also resulted in diminished myosin content. Conclusions: Our novel findings show that extracellular 14-3-3 proteins may act as previously unidentified myokines and may signal via CD13 to help maintain muscle mass

Numerical Construction of LISS Lyapunov Functions under a Small Gain Condition

In the stability analysis of large-scale interconnected systems it is frequently desirable to be able to determine a decay point of the gain operator, i.e., a point whose image under the monotone operator is strictly smaller than the point itself. The set of such decay points plays a crucial role in checking, in a semi-global fashion, the local input-to-state stability of an interconnected system and in the numerical construction of a LISS Lyapunov function. We provide a homotopy algorithm that computes a decay point of a monotone op- erator. For this purpose we use a fixed point algorithm and provide a function whose fixed points correspond to decay points of the monotone operator. The advantage to an earlier algorithm is demonstrated. Furthermore an example is given which shows how to analyze a given perturbed interconnected system.Comment: 30 pages, 7 figures, 4 table

Praise or blame? Affective influences on attributions for achievement

Three experiments showed that mood influences achievement attributions and that cognitive processes underlie these effects. In Experiment 1, happy Ss made more internal and stable attributions for success than failure in typical 'life dilemmas'. In Experiment 2, attributions for real-life exam performance were more internal and stable in a happy than in a sad mood. Dysphoric moods resulted in self-critical rather than self-enhancing attributions, contrary to motivational theories, but consistent with cognitive models and the clinical literature on depression. In Experiment 3 this pattern was repeated with direct self vs. other comparisons, and for self-efficacy judgments. The results are interpreted as supporting cognitive rather than motivational theories of attribution biases. The implications of the results for clinical research, and contemporary affect-cognition theories are considered

On the problem of interactions in quantum theory

The structure of representations describing systems of free particles in the theory with the invariance group SO(1,4) is investigated. The property of the particles to be free means as usual that the representation describing a many-particle system is the tensor product of the corresponding single-particle representations (i.e. no interaction is introduced). It is shown that the mass operator contains only continuous spectrum in the interval $(-\infty,\infty)$ and such representations are unitarily equivalent to ones describing interactions (gravitational, electromagnetic etc.). This means that there are no bound states in the theory and the Hilbert space of the many-particle system contains a subspace of states with the following property: the action of free representation operators on these states is manifested in the form of different interactions. Possible consequences of the results are discussed.Comment: 35 pages, Late

Could Only Fermions Be Elementary?

In standard Poincare and anti de Sitter SO(2,3) invariant theories, antiparticles are related to negative energy solutions of covariant equations while independent positive energy unitary irreducible representations (UIRs) of the symmetry group are used for describing both a particle and its antiparticle. Such an approach cannot be applied in de Sitter SO(1,4) invariant theory. We argue that it would be more natural to require that (*) one UIR should describe a particle and its antiparticle simultaneously. This would automatically explain the existence of antiparticles and show that a particle and its antiparticle are different states of the same object. If (*) is adopted then among the above groups only the SO(1,4) one can be a candidate for constructing elementary particle theory. It is shown that UIRs of the SO(1,4) group can be interpreted in the framework of (*) and cannot be interpreted in the standard way. By quantizing such UIRs and requiring that the energy should be positive in the Poincare approximation, we conclude that i) elementary particles can be only fermions. It is also shown that ii) C invariance is not exact even in the free massive theory and iii) elementary particles cannot be neutral. This gives a natural explanation of the fact that all observed neutral states are bosons.Comment: The paper is considerably revised and the following results are added: in the SO(1,4) invariant theory i) the C invariance is not exact even for free massive particles; ii) neutral particles cannot be elementar

Diquat Derivatives: Highly Active, Two-Dimensional Nonlinear Optical Chromophores with Potential Redox Switchability

In this article, we present a detailed study of structure−activity relationships in diquaternized 2,2′-bipyridyl (diquat) derivatives. Sixteen new chromophores have been synthesized, with variations in the amino electron donor substituents, π-conjugated bridge, and alkyl diquaternizing unit. Our aim is to combine very large, two-dimensional (2D) quadratic nonlinear optical (NLO) responses with reversible redox chemistry. The chromophores have been characterized as their PF_6^− salts by using various techniques including electronic absorption spectroscopy and cyclic voltammetry. Their visible absorption spectra are dominated by intense π → π^* intramolecular charge-transfer (ICT) bands, and all show two reversible diquat-based reductions. First hyperpolarizabilities β have been measured by using hyper-Rayleigh scattering with an 800 nm laser, and Stark spectroscopy of the ICT bands affords estimated static first hyperpolarizabilities β_0. The directly and indirectly derived β values are large and increase with the extent of π-conjugation and electron donor strength. Extending the quaternizing alkyl linkage always increases the ICT energy and decreases the E_(1/2) values for diquat reduction, but a compensating increase in the ICT intensity prevents significant decreases in Stark-based β_0 responses. Nine single-crystal X-ray structures have also been obtained. Time-dependent density functional theory clarifies the molecular electronic/optical properties, and finite field calculations agree with polarized HRS data in that the NLO responses of the disubstituted species are dominated by ‘off-diagonal’ β_(zyy) components. The most significant findings of these studies are: (i) β_0 values as much as 6 times that of the chromophore in the technologically important material (E)-4′-(dimethylamino)-N-methyl-4-stilbazolium tosylate; (ii) reversible electrochemistry that offers potential for redox-switching of optical properties over multiple states; (iii) strongly 2D NLO responses that may be exploited for novel practical applications; (iv) a new polar material, suitable for bulk NLO behavior

Evolution of Linear Absorption and Nonlinear Optical Properties in V-Shaped Ruthenium(II)-Based Chromophores

In this article, we describe a series of complexes with electron-rich cis-{Ru^(II)(NH_3)_4}^(2+) centers coordinated to two pyridyl ligands bearing N-methyl/arylpyridinium electron-acceptor groups. These V-shaped dipolar species are new, extended members of a class of chromophores first reported by us (Coe, B. J. et al. J. Am. Chem. Soc. 2005, 127, 4845−4859). They have been isolated as their PF_6− salts and characterized by using various techniques including ^1H NMR and electronic absorption spectroscopies and cyclic voltammetry. Reversible Ru^(III/II) waves show that the new complexes are potentially redox-switchable chromophores. Single crystal X-ray structures have been obtained for four complex salts; three of these crystallize noncentrosymmetrically, but with the individual molecular dipoles aligned largely antiparallel. Very large molecular first hyperpolarizabilities β have been determined by using hyper-Rayleigh scattering (HRS) with an 800 nm laser and also via Stark (electroabsorption) spectroscopic studies on the intense, visible d → π^* metal-to-ligand charge-transfer (MLCT) and π → π^* intraligand charge-transfer (ILCT) bands. The latter measurements afford total nonresonant β_0 responses as high as ca. 600 × 10^(−30) esu. These pseudo-C_(2v) chromophores show two substantial components of the β tensor, β_(zzz) and β_(zyy), although the relative significance of these varies with the physical method applied. According to HRS, β_(zzz) dominates in all cases, whereas the Stark analyses indicate that β_(zyy) is dominant in the shorter chromophores, but β_(zzz) and β_(zyy) are similar for the extended species. In contrast, finite field calculations predict that β_(zyy) is always the major component. Time-dependent density functional theory calculations predict increasing ILCT character for the nominally MLCT transitions and accompanying blue-shifts of the visible absorptions, as the ligand π-systems are extended. Such unusual behavior has also been observed with related 1D complexes (Coe, B. J. et al. J. Am. Chem. Soc. 2004, 126, 3880−3891)