Search

Porcine notochordal cell-derived matrix (NCM) has anti-inflammatory and regenerative effects on degenerated intervertebral discs. For its clinical use, safety must be assured. The porcine DNA is concerning because of (1) the transmission of endogenous retroviruses and (2) the inflammatory potential of cell-free DNA. Here, we present a simple, detergent-free protocol: tissue lyophilization lyses cells, and matrix integrity is preserved by limiting swelling during decellularization. DNA is digested quickly by a high nuclease concentration, followed by a short washout. Ninety-four percent of DNA was removed, and there was no loss of glycosaminoglycans or collagen. Forty-three percent of the total proteins remained in the decellularized NCM (dNCM). dNCM stimulated as much GAG production as NCM in nucleus pulposus cells but lost some anti-inflammatory effects. Reconstituted pulverized dNCM yielded a soft, shear-thinning biomaterial with a swelling ratio of 350% that also acted as an injectable cell carrier (cell viabilitygt;70%). dNCM can therefore be used as the basis for future biomaterials aimed at disc regeneration on a biological level and may restore joint mechanics by creating swelling pressure within the intervertebral disc.

PDF - 148K, Fig S4. Effect of PKD inhibitor on clonal formation.

PDF - 142K, Fig. S2. PKD1, PKD2 and PKD3 mRNA expression in cell lines from the Cancer Cell Line Encyclopedia database.

PDF - 190K, Fig. S3. Effect of kb-NB142-70 on RKO cell cycle, apoptosis, and NF-κB activity.

Protein kinase D (PKD) signaling plays a critical role in the regulation of DNA synthesis, proliferation, cell survival, adhesion, invasion/migration, motility, and angiogenesis. To date, relatively little is known about the potential role of PKD in the development and/or progression of human colorectal cancer. We evaluated the expression of different PKD isoforms in colorectal cancer and investigated the antitumor activity of PKD inhibitors against human colorectal cancer. PKD2 was the dominant isoform expressed in human colon cancer cells. PKD3 expression was also observed but PKD1 expression, at both the RNA and protein levels, was not detected. Suppression of PKD using the small molecule inhibitors CRT0066101 and kb-NB142-70 resulted in low micromolar in vitro antiproliferative activity against multiple human colorectal cancer cell lines. Drug treatment was associated with dose-dependent suppression of PKD2 activation. Incubation with CRT0066101 resulted in G2–M phase arrest and induction of apoptosis in human colorectal cancer cells. Further studies showed that CRT0066101 treatment gave rise to a dose-dependent increase in expression of cleaved PARP and activated caspase-3, in addition to inhibition of AKT and ERK signaling, and suppression of NF-κB activity. Transfection of PKD2-targeted siRNAs resulted in similar effects on downstream pathways as observed with small molecule inhibitors. Daily administration of CRT0066101 resulted in significant inhibition of tumor growth in HCT116 xenograft nude mice. Taken together, our studies show that PKD plays a significant role in mediating growth signaling in colorectal cancer and may represent a novel chemotherapeutic target for the treatment of colorectal cancer. Mol Cancer Ther; 13(5); 1130–41. ©2014 AACR.

PDF - 54K, Fig. S1. PKD1, PKD2 and PKD3 mRNA expression in colon cancer cells lines from the CellMiner database.

Purpose: Raf-kinases include three major isoforms. Although the role of B-Raf in melanoma is well established, little is known about C-Raf. We studied effects of C-Raf knockdown in vitro and assessed expression of C-Raf in a large cohort of melanomas and nevi.Experimental Design: Using specific siRNAs, we knocked down C-Raf expression, and determined the effect on viability, MAP extracellular signal-regulated kinase (ERK)/ERK kinase signaling, and apoptosis in seven melanoma cell lines. We determined the IC50 of the C-Raf inhibitors sorafenib and GW5074, and studied the effects of GW5074 on cell signaling. Using an automated method to measure in situ protein expression, we quantified C-Raf expression in 263 nevi and 523 melanomas.Results: C-Raf was knocked down in three cell lines with detectable phospho-C-Raf, resulting in decreased viability in two of the three (YULAC and YUROB). This resulted in decreased Bcl-2 expression and phospho-Bad cleavage, without affecting phospho-MEK and phospho-ERK. Sensitivity to sorafenib and GW5074 varied. GW5074 inhibited mitogen-activated protein kinase signaling without Bcl-2 and phospho-Bad down-regulation. C-Raf was highly expressed in melanomas compared with nevi (P < 0.0001), and no nevi had high C-Raf expression. C-Raf expression was higher in metastatic than primary specimens (P = 0.0225).Conclusions: C-Raf siRNA knock-down results in decreased viability of YULAC (B-RafV600K) and YUROB (B-RafWT) melanoma cells, likely mediated by Bcl-2 inhibition rather than mitogen-activated protein kinase inhibition. Cotargeting C-Raf and parallel pathways might be an effective therapeutic approach for melanoma. C-Raf expression is up-regulated in a subset of melanomas but not in nevi, suggesting that it might be a valuable diagnostic marker and therapeutic target. (Clin Cancer Res 2009;15(18):5704–13)

Supplementary Data from C-Raf Is Associated with Disease Progression and Cell Proliferation in a Subset of Melanomas

Zusammenfassung Hintergrund Sprachtherapeutisch-linguistische Fachkreise empfehlen die Anpassung einer von einem internationalen Konsortium empfohlenen Änderung der Nomenklatur für Sprachstörungen im Kindesalter, insbesondere für Sprachentwicklungsstörungen (SES), auch für den deutschsprachigen Raum. Fragestellung Ist eine solche Änderung in der Terminologie aus ärztlicher und psychologischer Sicht sinnvoll? Material und Methode Kritische Abwägung der Argumente für und gegen eine Nomenklaturänderung aus medizinischer und psychologischer Sicht eines Fachgesellschaften- und Leitliniengremiums. Ergebnisse Die ICD-10-GM (Internationale statistische Klassifikation der Krankheiten und verwandter Gesundheitsprobleme, 10. Revision, German Modification) und eine S2k-Leitlinie unterteilen SES in umschriebene SES (USES) und SES assoziiert mit anderen Erkrankungen (Komorbiditäten). Die USES- wie auch die künftige SES-Definition der ICD-11 (International Classification of Diseases 11th Revision) fordern den Ausschluss von Sinnesbehinderungen, neurologischen Erkrankungen und einer bedeutsamen intellektuellen Einschränkung. Diese Definition erscheint weit genug, um leichtere nonverbale Einschränkungen einzuschließen, birgt nicht die Gefahr, Kindern Sprach- und weitere Therapien vorzuenthalten und erkennt das ICD(International Classification of Disease)-Kriterium, nach dem der Sprachentwicklungsstand eines Kindes bedeutsam unter der Altersnorm und unterhalb des seinem Intelligenzalter angemessenen Niveaus liegen soll, an. Die intendierte Ersetzung des Komorbiditäten-Begriffs durch verursachende Faktoren, Risikofaktoren und Begleiterscheinungen könnte die Unterlassung einer dezidierten medizinischen Differenzialdiagnostik bedeuten. Schlussfolgerungen Die vorgeschlagene Terminologie birgt die Gefahr, ätiologisch bedeutsame Klassifikationen und differenzialdiagnostische Grenzen zu verwischen und auf wertvolles ärztliches und psychologisches Fachwissen in Diagnostik und Therapie sprachlicher Störungen im Kindesalter zu verzichten.

Background Cell-to-cell variation in gene expression strongly affects population behavior and is key to multiple biological processes. While codon usage is known to affect ensemble gene expression, how codon usage influences variation in gene expression between single cells is not well understood. Results Here, we used a Sort-seq based massively parallel strategy to quantify gene expression variation from a green fluorescent protein (GFP) library containing synonymous codons in Escherichia coli. We found that sequences containing codons with higher tRNA Adaptation Index (TAI) scores, and higher codon adaptation index (CAI) scores, have higher GFP variance. This trend is not observed for codons with high Normalized Translation Efficiency Index (nTE) scores nor from the free energy of folding of the mRNA secondary structure. GFP noise, or squared coefficient of variance (CV2), scales with mean protein abundance for low-abundant proteins but does not change at high mean protein abundance. Conclusions Our results suggest that the main source of noise for high-abundance proteins is likely not originating at translation elongation. Additionally, the drastic change in mean protein abundance with small changes in protein noise seen from our library implies that codon optimization can be performed without concerning gene expression noise for biotechnology applications.

Targeted therapies, such as EGFR inhibitors, have improved survival for patients with non-small cell lung cancer (NSCLC). Overall cure rates remain low due to eventual development of resistance. Osimertinib is a third-generation EGFR inhibitor that overcomes T790M-mediated resistance to earlier generation EGFRi. However, resistance to osimertinib has also emerged through mechanisms that are EGFR-dependent and EGFR-independent. To uncover novel strategies to overcome osimertinib resistance, we explored androgen receptor (AR) antagonists in combination with osimertinib as recent studies have demonstrated AR inhibition enhanced TKI-targeted therapy. For these experiments, we utilized lab-derived osimertinib-resistant PC9-AR and HCC827-AR NSCLC cell lines and matching parental lines in both in vitro and in vivo model systems. Clonogenic assays revealed that darolutamide in combination with osimertinib synergistically suppressed clonal growth in resistant cell lines. Similar results were observed with the combination of enzalutamide and osimertinib. Given this result, we next wanted to establish whether the promising in vitro activity of this combination translates into antitumor efficacy in vivo. Mice bearing either HCC827AR or PC9AR xenografts were treated orally daily with both drugs. At the doses administered, neither drug alone had antitumor activity. However, the combination significantly delayed tumor growth by more than 60%. We confirmed this ability to reverse resistance as a class effect using a different AR antagonist, enzalutamide in PC9AR-bearing mice. Significant tumor growth inhibition was observed with the combination of enzalutamide and osimertinib. To investigate the mechanism by which this combination may reverse osimertinib-resistance, RNA-seq studies were performed following drug exposure in vitro using parental and resistant cell lines under various experimental conditions. Comparison of gene expression profiles between parental and resistant cell lines under different therapeutic pressure and pathway enrichment analysis revealed important differentially expressed genes pointing plausible activated pathways that underline osimertinib resistance. AR antagonists have the potential to reverse EGFR inhibitor resistance in treatment of EGFR-mutant NSCLC. Citation Format: John C. Schmitz, Guojing Zhang, Zhentao Liu, Yufei Huang, Gabriel Sica, Shi-Yong Sun, Taofeek K. Owonikoko. Darolutamide reverses osimertinib resistance in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1752.

Small cell lung cancer (SCLC) is an aggressive neuroendocrine tumor characterized by rapid disease progression and poor patient survival. Key transcription factors implicated as drivers of unique biological phenotypes of SCLC include ASCL1, NEUROD1, YAP1, and POU2F3. Strategies to exploit these phenotypes for innovative precision medicine approaches will be impactful. Using an unbiased agnostic preclinical drug screen to uncover therapeutic opportunities in SCLC, we identified a strong signal with PLK1 inhibitors (PLK1i) with low nanomolar IC50. We extended the in vitro findings by testing the efficacy of PLK1i in vivo using traditional xenograft of SCLC H526 cell line and patient derived xenografts (PDX). Volasertib achieved significant tumor growth inhibition relative to control in H526 xenografts. Also, onvansertib significantly inhibited growth of platinum-resistant and platinum-sensitive PDXs. The combination of PLK1i with standard chemotherapeutic agents identified promising synergy of the combination of onvansertib and paclitaxel. We further interrogated for predictive biomarkers of PLK1i sensitivity using gene expression profile comparing highly sensitive to less sensitive cell lines. High expression of C-MYC but not PLK1, TP53 or RB was associated with resistance to PLK1i. Conversely, while TP53 expression level did not correlate, TP53 gene mutation status (inactivating disruptive mutations) correlated with cell sensitivity to PLK1i. We queried the publicly available CCLE and the Cancer Therapeutics Response Portal to evaluate whether any of the SCLC subtypes have therapeutic vulnerability to PLK1i. In general, high expression of YAP1 in SCLC cell lines correlated with greater sensitivity to PLK1i. A YAP1 positive cell line, SW1271, with strong TP53 expression was particularly resistant to PLK1i. CRISPR knockout of YAP1 in this cell line enhanced SW1271 sensitivity to PLK1i suggesting that YAP1 expression as a marker of vulnerability to PLK1i could be context dependent especially when co-occurring with TP53 mutations. The mechanism of this interaction will be discussed. Citation Format: John C. Schmitz, Guojing Zhang, Andrey A. Ivanov, Abbe Pannucci, Maya Ridinger, Taofeek K. Owonikoko. Targeting PLK1 effectively suppresses growth of small cell lung cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4995.

Metabolic engineering has allowed the production of a diverse number of valuable chemicals using microbial organisms. Many biological challenges for improving bio-production exist which limit performance and slow the commercialization of metabolically engineered systems. Dynamic metabolic engineering is a rapidly developing field that seeks to address these challenges through the design of genetically encoded metabolic control systems which allow cells to autonomously adjust their flux in response to their external and internal metabolic state. This review first discusses theoretical works which provide mechanistic insights and design choices for dynamic control systems including two-stage, continuous, and population behavior control strategies. Next, we summarize molecular mechanisms for various sensors and actuators which enable dynamic metabolic control in microbial systems. Finally, important applications of dynamic control to the production of several metabolite products are highlighted, including fatty acids, aromatics, and terpene compounds. Altogether, this review provides a comprehensive overview of the progress, advances, and prospects in the design of dynamic control systems for improved titer, rate, and yield metrics in metabolic engineering.

Background and ObjectivesThis study tested the hypothesis that treatment of myofascial trigger points (MTrPs) in the upper trapezius muscle (UTM) with repeated injection of 1% lidocaine results in better alleviation of muscular stiffness and soreness as well as improved metabolism in the hypercontracted MTrP area than treatment with radial extracorporeal shock wave therapy (rESWT).Materials and MethodsA single-blinded, prospective, randomized controlled trial was conducted on patients suffering from MTrPs in the UTM. Thirty patients were treated with repeated injection of 2 ml of 1% lidocaine (three injections; one injection per week). Another 30 patients were treated with rESWT (three treatment sessions; one treatment session per week; 2000 radial extracorporeal shock waves per treatment session; positive energy flux density = 0.10 mJ/mm2). The primary outcome measure was pain severity using the VAS score. The secondary outcome measures included muscle elasticity index, pressure pain threshold and neck disability index. Evaluation was performed at baseline (T1), 15-30 minutes after the first treatment in order to register immediate treatment effects (T2), before the second treatment (i.e., one week after baseline) (T3) and one week after the third treatment (i.e., four weeks after baseline) (T4).ResultsThere were no statistically significant differences in the primary and secondary outcome measures between the patients in the lidocaine arm and the patients in the rESWT arm at T1 and T4. Within the arms, the mean differences of all outcomes were statistically significant (P < 0.001) when comparing the data obtained at T1 with the data obtained at T3 and the data obtained at T4.ConclusionsThe results of this pilot study suggest that the use of rESWT in patients with MTrPs in the UTM is safe and leads to reduced pain and improved muscle elasticty, pressure pain threshold and neck disability index, without adverse effects. Larger trials are necessary to verify this. Clinicians should consider rESWT instead of injections of lidocaine in the treatment of MTrPs in the UTM.

A reaction-based sensor (NAS-1) showed a high affinity and sensitivity to HSO3- via a nucleophilic addition reaction in the aqueous media, giving dual signals from absorption and emission spectra. NAS-1 was successfully applied in RK13 epithelial cells to detect HSO3- in a cellular environment.

Magnetite-binding proteins are in high demand for the functionalization of magnetic nanoparticles. Binding analysis of six previously uncharacterized proteins from the magnetotactic Deltaproteobacterium Desulfamplus magnetovallimortis BW-1 identified two new magnetite-binding proteins (Mad10, Mad11). These proteins can be utilized as affinity tags for the immobilization of recombinant fusion proteins to magnetite.

Magnetite-binding proteins are in high demand for the functionalization of magnetic nanoparticles. Binding analysis of six previously uncharacterized proteins from the magnetotactic

PURPOSE: We investigated the combination of tivantinib, a c-MET tyrosine kinase inhibitor (TKI), and bevacizumab, an anti-VEGF-A antibody. METHODS: Patients with advanced solid tumors received bevacizumab (10 mg/kg intravenously every 2 weeks) and escalating doses of tivantinib (120–360 mg orally twice daily). In addition to safety and preliminary efficacy, we evaluated pharmacokinetics of tivantinib and its metabolites, as well as pharmacodynamic biomarkers in peripheral blood and skin. RESULTS: Eleven patients received the combination treatment, which was generally well tolerated. The main dose-limiting toxicity was grade 3 hypertension, which was observed in four patients. Other toxicities included lymphopenia and electrolyte disturbances. No exposure-toxicity relationship was observed for tivantinib or metabolites. No clinical responses were observed. Mean levels of the serum cytokine bFGF increased (p = 0.008) after the bevacizumab-only lead-in and decreased back to baseline (p = 0.047) after addition of tivantinib. Tivantinib reduced levels of both phospho-MET (7/11 patients) and tubulin (4/11 patients) in skin. CONCLUSIONS: The combination of tivantinib and bevacizumab produced toxicities that were largely consistent with the safety profiles of the individual drugs. The study was terminated prior to establishment of the recommended phase II dose (RP2D) due to concerns regarding the mechanism of tivantinib, as well as lack of clinical efficacy seen in this and other studies. Tivantinib reversed the upregulation of bFGF caused by bevacizumab, which has been considered a potential mechanism of resistance to therapies targeting the VEGF pathway. The findings from this study suggest that the mechanism of action of tivantinib in humans may involve inhibition of both c-MET and tubulin expression. TRIAL REGISTRATION: NCT01749384 (First posted 12/13/2012).

Colloidal semiconductor quantum dots/graphene van der Waals (vdW) heterojunctions take advantages of the enhanced light-matter interaction and spectral tenability of quantum dots (QDs) and superior charge mobility in graphene, providing a promising alternative for uncooled infrared photodetectors with a gain or external quantum efficiency up to 1010. In these QD/graphene vdW heterostructures, the QD/graphene interface plays a critical role in controlling the optoelectronic process including exciton dissociation, charge injection and transport. Specifically, charge traps at the vdW interface can increase the noise, reduce the responsivity and response speed. This paper highlight our recent progress in engineering the vdW heterojunction interface towards more efficient charge transfer for higher photoresponsivity, D* and response speed. These results illustrate that the importance in vdW heterojunction interface engineering in QD/graphene photodetectors which may provide a promising pathway for low-cost, printable and flexible infrared detectors and imaging systems.

The SNAP-27 (Systems Nuclear Auxiliary Power) was a radioisotope thermoelectric generator (RTG) designed and developed to operate in the adverse lunar surface environment for periods of greater than one year, and considered by some to be the pathfinder for what would become the modern RTG program. This RTG provided approximately 70 watts of electrical power to the Apollo Lunar Surface Experiment Package (ALSEP) during both lunar day and night conditions. The first SNAP-27 became operational on the moon in November, 1969, during Apollo 12. SNAP-27s also flew on all of the subsequent Apollo missions, and each unit that was deployed on the moon operated for five to eight years until the ALSEP stations were shut down. While the SNAP-27 performance was not as remarkable as later RTG designs (e.g. the marquee Voyager missions), it is important to note that all of the deployed units exceeded their one/two-year design lifetimes. Impressive by today's standards, the development of the SNAP-27 was initiated only 53-months before the launch of Apollo 12. Review of the available literature from General Electric (GE), NASA, and the Atomic Energy Commission (AEC) reveals that a very extensive design, development, and testing program was conducted prior to the launch of Apollo 12. Ground testing comprised of couple-, module-, and system-level testing to characterize the behavior of the unit to the rigors of the anticipated lunar environment. Over 50 years after those historic missions, the SNAP-27 program much like the science data returned from the ALSEP stations still has something to teach us. As NASA continues to plan for sustained lunar exploration, key capabilities will be identified and developed to establish the first long-term foothold on the lunar frontier. This paper identifies lessons that can be applied from the SNAP-27 program to address the technical challenges/risks to employing current (e.g. MMRTG) and future RTG systems on the lunar surface.

The Multi-Mission Radioisotope Thermoelectric Generator (MMRTG) is a readily available, long-lived, small scale, versatile power supply that can generate uninterrupted power on the moon during the long lunar nights. Lunar surface conditions, however, can be very harsh. Lunar daytime highs could cause the critical hot junction temperature (T hj ) to exceed its operating limits, while the cold lunar nights can generate deep thermal cycles that could induce significant stress. An energy balance for objects on the lunar surface was performed, and an approximate lunar thermal sink for the MMRTG was obtained. This approximate sink does not incorporate potential self-shading from incident solar radation created by the MMRTG geometry. It is hypothesized, however, that the effect of this self-shading on the lunar surface will result in a decrease in operational temperatures. Recently developed thermal models have been improved so that they can calculate critical temperatures for an MMRTG on the lunar surface. Using the approximate lunar thermal sink, results from the model indicate that under the nominal operating conditions of 28 V and 2000 W th of thermal inventory, the MMRTG can operate on the lunar surface while remaining under the maximum T hj by a margin of greater than 25 °C. Operating under worst-case nominal conditions, 28 V and 2048 W th of thermal inventory, still allows operations on the lunar surface with over 15 °C of margin. Operating at higher voltages will increase temperatures within the MMRTG, while lower voltages are expected to cause T hj to decrease. An off-nominal 32 V system will still have at least 15 °C of margin if the system can be held at 2000 W th or less of thermal inventory, but that margin shrinks to almost zero if the maximum specification of 2048 W th happens to be implemented. Lowering the thermal inventory can lower T hj , but it will also decrease beginning-of-life power. The relationship between these three parameters appears to be linear, with a 48 W th decrease in thermal inventory causing T hj and beginning-of-life power to decrease by 11.2 °C and 4.8 We, respectively. Missions that want to use MMRTG on the lunar surface should carefully consider the voltage they want to pull from the generator. If the operating voltage chosen is high, then a decrease in thermal inventory is one potential option for keeping the T hj below the operational maximum. Results also indicate that the MMRTG will experience a thermal cycle of $\Delta\mathrm{T}=47\ {}^{\circ}\mathrm{C}$ during the lunar diurnal cycle. Empirical testing of a prototypic system or subsystem is anticipated to be the best method for determining the effect of lunar induced thermal cycles on the MMRTG. Currently, the Engineering Unit is available and is not a critical testing resource for NASA. As such, potential damage to the unit from thermal cycle testing would not deprive NASA of valuable life test data on the MMRTG. Other thermal cycle testing options include an MMRTG thermoelectric module mounted in a suitable testing frame.

Colloidal semiconductor quantum dot/graphene van der Waals heterostructure nanohybrids are emerging technologies for photodetection. These nanohybrids combine the advantages of the enhanced light-matter interaction and spectral tunability of quantum dots (QDs) and superior charge mobility in graphene, providing an affordable alternative for uncooled photodetectors with high gain or external quantum efficiency in a wide spectral range. In particular, the interfacing of QDs with high-mobility graphene as a charge transport pathway has provided an effective resolution to the critical issue of charge transport in QD-only photodetectors stemming from the low charge mobility associated with both QD surface defect states and QD-QD junctions. Furthermore, the achieved capability in industrial-scale fabrication of graphene, colloidal QDs, and QD/graphene nanohybrids has motivated efforts in researches of focal plane arrays that are expected to be not only high performance and low cost but also lightweight, flexible, and wearable. This chapter aims to provide an up-to-date review of the recent progress made in the research of QD/graphene nanohybrid photodetectors, together with a discussion on the challenges remaining and perspective in future research and development to make QD/graphene nanohybrids competitive for commercialization.

Decellularization of animal tissues is a novel route to obtain biomaterials for use in tissue engineering and organ transplantation. Successful decellularization is required as animal DNA causes inflammatory reactions and contains endogenous retroviruses, which could be transmitted to the patient. One of the criteria for successful decellularization is digestion (fragmentation) and elimination (residual quantity) of DNA from the tissue. Quantification of DNA can be done in many ways, but it has recently been shown that silica-based solid-phase extraction methods often do not completely purify in particular small DNA fragments. In the context of decellularization, this means that the measured DNA amount is underestimated, which could compromise safety of the processed tissue for in-patient use. In this article, we review DNA quantification methods used by researchers and assess their influence on the reported DNA contents after decellularization. We find that underestimation of residual DNA amount after silica-based solid-phase extraction may be as large as a factor of ten. We therefore recommend a direct assessment of DNA amount in tissue lysate using dsDNA-specific binding dyes, such as Picogreen, due to their higher accuracy for small fragment detection as well as ease of use and widespread availability.

A reaction-based sensor (NAS-1) showed a high affinity and sensitivity to HSO3- via a nucleophilic addition reaction in the aqueous media, giving dual signals from absorption and emission spectra. NAS-1 was successfully applied in RK13 epithelial cells to detect HSO3- in a cellular environment.

The successful testing of the Kilopower reactor during the KRUSTY (Kilopower Reactor Using Stirling TechnologY) experiment significantly reduced the risk to fly fission power systems by demonstrating stable reactor operation through nominal and severe simulated mission scenarios. The experiment validated the neutronics, heat transfer, and power conversion systems needed for 1 kilowatt of electrical power production from the Kilopower reactor. The need for higher power reactors to support human exploration missions to the moon and Mars has become increasingly important due to the urgency to put boots on the moon by 2024 and have a sustainable presence in the following years. This desire has prompted NASA to continue the development of the Kilopower reactor to extend the power up to 10 kilowatts of electricity in support of a lunar base. These 10 kilowatt units are expected to be used as standalone units or be ganged together to create a modular power grid for propellent production, human habitats, and robotic exploration to name a few. The Kilopower reactor was originally designed to produce electrical power from 1 to 10 kilowatts using the same highly enriched uranium fuel, sodium heat pipes, and Stirling convertors at the proper scale. Consideration has also been given to the use of low enriched uranium fuel for these missions and will be studied along with the other aspects of the reactor. This paper will focus on the design concepts and trades associated with the scale up of the Kilopower power conversion system and heat transfer system to support human exploration of the moon and Mars.

We present a preliminary design for a 3 kg expendable measurement system to descend to a depth of up to 1 km in the cryogenic hydrocarbon seas (Kraken and Ligeia Mare) of Saturn’s moon Titan. These could profile the temperature and gross composition (methane/ethane) and providing information on the constitution of the seabed. Particular features of the Titan environment distinct from Earth are (1) the interior of this small vehicle must be thermally isolated from the 94 K liquid (2) 1 km depth on Titan entails overpressures of only 10 bar, and (3) that the low dielectric loss of the liquid allows data transmission by radio link. A surface vessel equipped with such expendable dropsondes could accomplish much of the science previously considered for submersible vehicles, while avoiding the challenges of buoyancy control.

Disorder plays an essential role in shaping the transport properties of GeSbTe phase-change materials (PCMs) to enable nonvolatile memory technology. Recently, increasing efforts have been undertaken to investigate disorder in the stable hexagonal phase of GeSbTe compounds, focusing on a special type of swapping bilayer defects. This configuration has been claimed to be the key element for a new mechanism for phase-change memory. Here, we report a direct atomic-scale chemical identification of these swapping bilayer defects in hexagonal GeSb2Te4 together with nanoscale atomic modeling and simulations. We identify the intermixing of Sb and Te in the bilayer to be the essential ingredient for the stability of the defects, and elucidate their abundance as due to the small energy cost. The bilayer defects are demonstrated to be ineffective in altering the electron localization nature that is relevant to transport properties of hexagonal GeSb2Te4. Our work paves the way for future studies of layer-switching dy...

Zusammenfassung. Fragestellung: Epidemiologische Studien weisen auf die Bedeutung von prä- und perinatalen Risikofaktoren bei der Entstehung der Aufmerksamkeitsdefizit-/Hyperaktivitätsstörung (ADHS) und der Autismus-Spektrum-Störung (ASS) hin. Die vorliegende Arbeit vergleicht in einer klinischen Stichprobe von Kindern mit ADHS, ASS, dem kombinierten Störungsbild sowie ADHS und Störung des Sozialverhaltens (SSV) mögliche störungsspezifische Risikofaktoren und untersucht, ob das Vorliegen mehrerer Risikofaktoren das Auftreten von kombinierten Störungsbildern begünstigt. Methodik: Prä- und perinatale Risikofaktoren von insgesamt 341 Patienten (299 Jungen, 42 Mädchen) aus einer klinischen Inanspruchnahmepopulation werden zwischen Kindern mit ADHS (N = 80), ASS (N = 122), ADHS & ASS (N = 55) oder ADHS & SSV (N = 84) verglichen. Ergebnisse: Die Mütter von Kindern mit ADHS & SSV rauchten häufiger in der Schwangerschaft, die Eltern wiesen einen niedrigeren Bildungsstatus und häufiger Migrationsstatus auf als von Kindern mit ASS oder ADHS. Kinder mit ASS hatten häufiger einen Migrationshintergrund als Kinder mit ADHS. Fehlgeburten fanden sich als spezifischer Risikofaktor für das kombinierte Störungsbild ADHS & ASS. Schlussfolgerung: Zahlreiche der in epidemiologischen Studien beschriebenen Risikofaktoren fanden sich in der klinischen Stichprobe selten. Die meisten Risikofaktoren lagen in derselben Ausprägung bei allen Krankheitsbildern vor.

Polymeric microspheres have been utilized in a broad range of applications ranging from chromatographic separation techniques to analysis of air flow over aerodynamic surfaces. The preparation of microspheres from many different polymer families has consequently been extensively studied using a variety of synthetic approaches. Although there are a variety of methods of synthesis for polymeric microspheres, free-radical initiated emulsion polymerization is one of the most common techniques. In this work, poly(styrene-co-methyl methacrylate) microspheres were synthesized via surfactant-free emulsion polymerization. The effect of co-monomer composition and addition time on particle size distribution, particle formation, and particle morphology were investigated. Particles were characterized using dynamic light scattering (DLS) and scanning electron microscopy (SEM) to gain further insight into particle size and size distributions. Reaction kinetics were analyzed alongside of characterization results. A particle formation mechanism for poly(styrene-co-methyl methacrylate) microspheres was proposed based on characterization results and known reaction kinetics.

Recently, the New Horizons spacecraft flew by the Kuiper Belt Object Ultima Thule 13 years after launch. While flybys are a necessary 'first look' at the object in question a more comprehensive evaluation of the object will require an orbiter. Due to these object’s low mass a chemical propulsion system is not a viable option for entering their shallow gravity wells. The Dawn mission showed that solar electric propulsion is significantly better at the task of reaching low mass objects, but at Kuiper belt distances solar power is not a viable choice. A small nuclear reactor based on the recent kilopower ground test could provide 1-10 kWe of power for an electric propulsion system. The NASA Compass Team developed a Nuclear Electric Propulsion Kuiper Belt Object Orbiter to explore what a vehicle would look like to orbit these deep space objects.

The academic study of the Phoenicians and their civilization began relatively recently, but has quickly generated a large bibliography of important studies. One of the difficulties that learners face is that the historical study of Phoenician civilization must draw on a large number of sources from several different fields of research, not all of which are in regular communication with each other or are widely known to the general readership. This chapter provides an introduction to and a sample of the many sources available to specialists and nonspecialists seeking reliable knowledge about Phoenician civilization from the Iron Age to the Hellenistic period. Terminology is introduced, beginning with designations of the land known as Canaan. Greek testimonies to Phoenician history are next presented, especially the figure Sanchuniathon discussed by Porphyry and Josephus. Phoenician alphabetic texts—that is, epigraphic sources—are discussed for their limited potential as historical sources, with ample bibliographic citations to guide the researcher.