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Introduction HGF/c-MET signaling is a significant driver of glioblastoma (GBM) growth and disease progression. Unfortunately, c-MET targeted therapies have been found to be largely ineffective suggesting additional redundant mechanisms of c-MET activation. Methods Utilizing RNA-sequencing (RNA-seq) and ribosome profiling analyses of circular RNAs, circ-HGF (hsa_circ_0080914) was identified as markedly upregulated in primary GBM and found to potentially encode an HGF protein variant (C-HGF) 119 amino acids in length. This candidate HGF variant was characterized and evaluated for its ability to mediate c-MET activation and regulate PDX GBM cell growth, motility and invasive potential in vitro and tumor burden in intracranial xenografts in mice. Results An internal ribosome entry site (IRES) was identified within the circ-HGF RNA which mediated translation of the cross-junctional ORF encoding C-HGF and was observed to be highly expressed in GBM relative to normal brain tissue. C-HGF was also found to be secreted from GBM cells and concentrated cell culture supernatants or recombinant C-HGF activated known signaling cascades downstream of c-MET. C-HGF was shown to interact directly with the c-MET receptor resulting in its autophosphorylation and activation in PDX GBM lines. Knockdown of C-HGF resulted in suppression of c-MET signaling and marked inhibition of cell growth, motility and invasiveness, whereas overexpression of C-HGF displayed the opposite effects. Additionally, modulation of C-HGF expression regulated tumor growth in intracranial xenografted PDX GBM models. Conclusions These results reveal an alternative mechanism of c-MET activation via a circular RNA encoded HGF protein variant which is relevant in GBM biology. Targeting C-HGF may offer a promising approach for GBM clinical management.

Heat shock proteins (HSPs), especially Hsp70 (HSPA1), have been associated with cellular protection from various cellular stresses including heat, hypoxia-ischemia, neurodegeneration, toxins, and trauma. Endogenous HSPs are often synthesized in direct response to these stresses but in many situations are inadequate in protecting cells. The present study addresses the transduction of Hsp70 into cells providing protection from acute oxidative stress by H2O2. The recombinant Fv-Hsp70 protein and two mutant Fv-Hsp70 proteins minus the ATPase domain and minus the ATPase and terminal lid domains were tested at 0.5 and 1.0 μM concentrations after two different concentrations of H2O2 treatment. All three recombinant proteins protected SH-SY5Y cells from acute H2O2 toxicity. This data indicated that the protein binding domain was responsible for cellular protection. In addition, experiments pretreating cells with inhibitors of antioxidant proteins catalase and gamma-glutamylcysteine synthase (GGCS) before H2O2 resulted in cell death despite treatment with Fv-Hsp70, implying that both enzymes were protected from acute oxidative stress after treatment with Fv-Hsp70. This study demonstrates that Fv-Hsp70 is protective in our experiments primarily by the protein-binding domain. The Hsp70 terminal lid domain was also not necessary for protection.

Heat shock proteins (HSPs), especially Hsp70 (HSPA1), have been associated with cellular protection from various cellular stresses including heat, hypoxia-ischemia, neurodegeneration, toxins, and trauma. Endogenous HSPs are often synthesized in direct response to these stresses but in many situations are inadequate in protecting cells. The present study addresses the transduction of Hsp70 into cells providing protection from acute oxidative stress by H2O2. The recombinant Fv-Hsp70 protein and two mutant Fv-Hsp70 proteins minus the ATPase domain, and minus the ATPase and terminal lid domains were tested at 0.5 and 1.0 uM concentrations after two different concentrations of H2O2treatment. All three recombinant proteins protected SH-SY5Y cells from acute H2O2toxicity. This data indicated that the protein binding domain was responsible for cellular protection. In addition, experiments pretreating cells with inhibitors of antioxidant proteins catalase and gamma-glutamylcysteine synthase (GGCS) before H2O2resulted in cell death despite treatment with Fv-Hsp70, implying that both enzymes were protected from acute oxidative stress after treatment with Fv-Hsp70. This study demonstrates that Fv-Hsp70 is protective in our experiments primarily by the protein-binding domain. The Hsp70 terminal lid domain was also not necessary for protection. Cellular protection was protective via the antioxidant proteins catalase and GGCS.

Cytoplasmic DNA triggers a cGAS-mediated signaling cascade that promotes an innate immune response and is potentially actionable in cancer immunotherapy. Here we show that a cytoplasmic-localizing lupus anti-DNA autoantibody activates cGAS and facilitates an immune-mediated prolongation of survival in orthotopic models of glioblastoma (GBM). Mechanistically, cellular penetration and blood-brain barrier crossing by the anti-DNA autoantibody is linked to nucleoside transport. Pulldown, knockdown, signaling, and cytotoxicity assays demonstrate autoantibody association with and activation of cGAS. In orthotopic GBM models, the autoantibody localizes to brain tumor, increases tumor CD8+ T cell content, and prolongs survival in immunocompetent but not immunodeficient mice. This work introduces the new concept of a cGAS-activating anti-DNA autoantibody, which impacts theories on mechanisms of autoimmunity and has translational applications in cancer immunotherapy.

The Hippo and mTOR signaling cascades are major regulators of cell growth and division. Aberrant regulation of these pathways has been demonstrated to contribute to gliomagenesis and result in enhanced glioblastoma proliferation and invasive characteristics. Several crosstalk mechanisms have been described between these two pathways, although a complete picture of these signaling interactions is lacking and is required for effective therapeutic targeting. Here we report the ability of mTORC2 to directly phosphorylate YAP at serine 436 (Ser436) positively regulating YAP activity. We show that mTORC2 activity enhances YAP transcriptional activity and the induction of YAP-dependent target gene expression while its ablation via genetic or pharmacological means has the opposite affects on YAP function. mTORC2 interacts with YAP via Sin1 and mutational analysis of serine 436 demonstrates that this phosphorylation event affects several properties of YAP leading to enhanced transactivation potential. Moreover, YAP serine 436 mutants display altered glioblastoma growth, migratory capacity and invasiveness both in vitro and in xenograft experiments. We further demonstrate that mTORC2 is able to regulate a Hippo pathway resistant allele of YAP suggesting that mTORC2 can regulate YAP independent of Hippo signaling. Correlative associations between the expression of these components in GBM patient samples also supported the presence of this signaling relationship. These results advance a direct mTORC2/YAP signaling axis driving GBM growth, motility and invasiveness.

Ξ− atomic X-ray spectroscopy is one of the most useful methods for investigation of the Ξ–nucleus strong interaction. Since the X-ray energy is shifted and/or broadened due to the Ξ–nucleus strong interaction compared to those calculated from electromagnetic interaction alone, the measurement of the energy shift, ΔE, and the width, Γ, give us information on the Ξ–nucleus potential. A serious problem in the measurement is the significant background derived from in-flight Ξ− decay. A novel method of identifying stopped Ξ− events using the nuclear emulsion was developed to realize the first Ξ− atomic X-ray spectroscopy experiment as the J-PARC E07 experiment, which also aimed at searching for ΛΛ and Ξ− hypernuclei in the emulsion. The X-rays emitted from Ξ− Br and Ξ− Ag atoms were measured using germanium detectors. No clear peaks were observed in the obtained spectra. However, we succeeded in reducing the background to 1/170 by this method employing coincidence measurements using nuclear emulsion and X-ray detectors.

The β-delayed one- and two-neutron emission probabilities (P_{1n} and P_{2n}) of 20 neutron-rich nuclei with N≥82 have been measured at the RIBF facility of the RIKEN Nishina Center. P_{1n} of ^{130,131}Ag, ^{133,134}Cd, ^{135,136}In, and ^{138,139}Sn were determined for the first time, and stringent upper limits were placed on P_{2n} for nearly all cases. β-delayed two-neutron emission (β2n) was unambiguously identified in ^{133}Cd and ^{135,136}In, and their P_{2n} were measured. Weak β2n was also detected from ^{137,138}Sn. Our results highlight the effect of the N=82 and Z=50 shell closures on β-delayed neutron emission probability and provide stringent benchmarks for newly developed macroscopic-microscopic and self-consistent global models with the inclusion of a statistical treatment of neutron and γ emission. The impact of our measurements on r-process nucleosynthesis was studied in a neutron star merger scenario. Our P_{1n} and P_{2n} have a direct impact on the odd-even staggering of the final abundance, improving the agreement between calculated and observed Solar System abundances. The odd isotope fraction of Ba in r-process-enhanced (r-II) stars is also better reproduced using our new data.

INTRODUCTION: Recent studies have suggested that dysregulated Hippo pathway signaling may contribute to glioblastoma proliferation and invasive characteristics. The downstream effector of the pathway, the Yes-associated protein (YAP) oncoprotein, has emerged as a promising target in glioblastoma multiforme (GBM). METHODS: Utilizing a high-throughput yeast two-hybrid based screen, a small molecule was identified which inhibits the association of the co-transcriptional activator YAP1 and the TEA domain family member 1 (TEAD1) transcription factor protein–protein interaction interface. This candidate inhibitor, NSC682769, a novel benzazepine compound, was evaluated for its ability to affect Hippo/YAP axis signaling and potential anti-glioblastoma properties. RESULTS: NSC682769 potently blocked association of YAP and TEAD in vitro and in GBM cells treated with submicromolar concentrations. Moreover, inhibitor-coupled bead pull down and surface plasmon resonance analyses demonstrate that NSC682769 binds to YAP. NSC682769 treatment of GBM lines and patient derived cells resulted in downregulation of YAP expression levels resulting in curtailed YAP-TEAD transcriptional activity. In GBM cell models, NSC682769 inhibited proliferation, colony formation, migration, invasiveness and enhanced apoptosis. In tumor xenograft and genetically engineered mouse models, NSC682769 exhibited marked anti-tumor responses and resulted in increased overall survival and displayed significant blood-brain barrier penetration. CONCLUSIONS: These results demonstrate that blockade of YAP-TEAD association is a viable therapeutic strategy for glioblastoma. On the basis of these favorable preclinical studies further clinical studies are warranted.

We investigate the impacts of neutrino cooling mechanism inside the neutron star (NS) core on the light curves of type-I X-ray bursts and X-ray superbursts. From several observations of NS thermal evolution, physical processes of fast neutrino cooling, such as the direct Urca (DU) process, are indicated. They significantly decrease the surface temperature of NSs, though the cooling effect could be suppressed by nucleon superfluidity. In the present study, focusing on the DU process and nucleon superfluidity, we investigate the effects of NS cooling on the X-ray bursts using a general-relativistic stellar-evolution code. We find that the DU process leads find the longer recurrence time and the higher peak luminosity, which could be obstructed by the neutrons superfluidity. We also apply our burst models to the comparison with {\it Clocked burster} GS 1826$-$24, and to the recurrence time of superburst triggered by carbon ignition. These effects are significant within a certain range of binary parameters and uncertainty of the NS equation of state., Comment: 12 pages, 8 figures, 3 Tables

INTRODUCTION: Clinical trials directed at mechanistic target of rapamycin (mTOR) inhibition have yielded disappointing results in glioblastoma (GBM). A major mechanism of resistance involves the activation of a salvage pathway stimulating internal ribosome entry site (IRES)-mediated protein synthesis. PRMT5 activity has been implicated in the enhancement of IRES activity. METHODS: We analyzed the expression and activity of PRMT5 in response to mTOR inhibition in GBM cell lines and short-term patient cultures. To determine whether PRMT5 conferred resistance we used genetic and pharmacological approaches to ablate PRMT5 activity and assessed the effects on in vitro and in vivo sensitivity. Mutational analyses of the requisite IRES-trans-acting factor (ITAF), hnRNP A1 determined whether PRMT5-mediated methylation was necessary for ITAF RNA binding and IRES activity. RESULTS: PRMT5 activity is stimulated in response to mTOR inhibitors. Knockdown or treatment with a PRMT5 inhibitor blocked IRES activity and sensitizes GBM cells. Ectopic expression of non-methylatable hnRNP A1 mutants demonstrated that methylation of either arginine residues 218 or 225 was sufficient to maintain IRES binding and hnRNP A1-dependent cyclin D1 or c-MYC IRES activity, however a double R218K/R225K mutant was unable to do so. The PRMT5 inhibitor EPZ015666 displayed synergistic anti-GBM effects in vitro and in a xenograft mouse model in combination with PP242. CONCLUSIONS: These results demonstrate that PRMT5 activity is stimulated upon mTOR inhibition in GBM. Our data further support a signaling cascade in which PRMT5-mediated methylation of hnRNP A1 promotes IRES RNA binding and activation of IRES-mediated protein synthesis and resultant mTOR inhibitor resistance.

We have performed the first direct measurement of the 83Rb(p,g) radiative capture reaction cross section in inverse kinematics using a radioactive beam of 83Rb at incident energies of 2.4 and 2.7 A MeV. The measured cross section at an effective relative kinetic energy of Ecm = 2.393 MeV, which lies within the relevant energy window for core collapse supernovae, is smaller than the prediction of statistical model calculations. This leads to the abundance of 84Sr produced in the astrophysical p process being higher than previously calculated. Moreover, the discrepancy of the present data with theoretical predictions indicates that further experimental investigation of p-process reactions involving unstable projectiles is clearly warranted., 6 pages, 4 figures, published in Physical Review Letters

Mosquitoes have developed specialized oviposition strategies that allow them to develop in a wide variety of aquatic habitats. Environmentally cued hatching traits may also play an important role in the successful colonization of some larval habitats, but this subject has remained largely unexplored in Culicidae. Aedes atropalpus (Coquillett) is an autogenous rock pool specialist that may maintain unique adaptations for oviposition and egg hatching. We investigated the egg-laying strategies of Ae. atropalpus exposed to standard (non-diapausing) rearing conditions and diapause-inducing conditions and tested the impact of physical agitation on egg hatch rates by exposing floating and submerged eggs to physical agitation treatments. The results of the oviposition experiment indicate that Ae. atropalpus females primarily lay non-diapausing eggs directly onto the water surface and lay diapausing eggs directly on solid surfaces. The egg-hatching experiment demonstrated that physical agitation significantly increases Ae. atropalpus hatch rates. Floating and submerged eggs responded similarly to the agitation treatment. These data suggest that oviposition behaviors based on both egg diapause status and environmentally-cued hatching strategies may be important adaptations for Ae. atropalpus in riverine rock pools.

The $\nu p$ process appears in proton-rich, hot matter which is expanding in a neutrino wind and may be realised in explosive environments such as core-collapse supernovae or in outflows from accretion disks. The impact of uncertainties in nuclear reaction cross sections on the finally produced abundances has been studied by applying Monte Carlo variation of all astrophysical reaction rates in a large reaction network. As the detailed astrophysical conditions of the $\nu p$ process still are unknown, a parameter study was performed, with 23 trajectories covering a large range of entropies and $Y_\mathrm{e}$. The resulting abundance uncertainties are given for each trajectory. The $\nu p$ process has been speculated to contribute to the light $p$ nuclides but it was not possible so far to reproduce the solar isotope ratios. It is found that it is possible to reproduce the solar $^{92}$Mo/$^{94}$Mo abundance ratio within nuclear uncertainties, even within a single trajectory. The solar values of the abundances in the Kr-Sr region relative to the Mo region, however, cannot be achieved within a single trajectory. They may still be obtained from a weighted superposition of different trajectories, though, depending on the actual conditions in the production site. For a stronger constraint of the required conditions, it would be necessary to reduce the uncertainties in the 3$\alpha$ and $^{56}$Ni(n,p)$^{56}$Co rates at temperatures $T>3$ GK., Comment: 6 pages, 1 figure, 2 tables; to appear in JPS Conference Proceedings (OMEG15, 2019); summarizes and expands on arXiv:1907.13129 with an additional discussion of isotopic ratios of p-nuclides; v2: typos fixed

Invasive container mosquito species Aedes aegypti (L.) and Ae. albopictus (Skuse) frequently co-occur in nature where they are typically segregated spatially by distinctive macrohabitat preferences that may select for different oviposition strategies. In a standard cage environment, we compared oviposition site selection by individual gravid females of these species exposed to variable numbers of water-holding cups and the presence or absence of a rugose wettable container lining. Offered plastic cups with and without lining, both species laid a significantly higher percentage of eggs in cups with lining (95.3% for Ae. aegypti, 88.4% for Ae. albopictus), than in cups without (4.7% for Ae. aegypti, 11.6% for Ae. albopictus). Linear regressions of container availability versus the number of lined cups colonized were similar for Ae. aegypti and Ae. albopictus. Rates of oviposition on the water surface differed by species for both cup types, with Ae. albopictus laying a higher percentage of eggs on the water surface in cups without lining (28.1% for Ae. aegypti, 51.3% for Ae. albopictus), than in cups with (5.3% for Ae. aegypti, 2.5% for Ae. albopictus). The more varied oviposition strategies of Ae. albopictus are interpreted in the context of its broader macrohabitat use and inferior egg desiccation resistance.

Ξ− atomic X-ray spectroscopy is one of the most useful methods for investigation of the Ξ-nucleus strong interaction. A serious problem in the measurement is the significant background coming from in-flight Ξ− decay. For the first Ξ− atomic X-ray spectroscopy experiment, a novel method of identifying stopped Ξ− events using nuclear emulsion was developed to reject background photons from in-flight Ξ− decay. We succeeded in reducing the background to 1/170 by this method employing coincidence measurements using the nuclear emulsion and X-ray detectors.

Rapid displacements of resident Aedes aegypti by invasions of Aedes albopictus have been documented in the southeastern United States and Bermuda. Interspecific mating has been detected in nature between these species and proposed as a likely mechanism for these displacements by means of asymmetric reproductive interference, or satyrization. However, rapid displacements of A. aegypti have not been detected in most localities where these two invasive species are known to co-occur. Aedes albopictus invaded the United States and Brazil at approximately the same time, in the mid-1980s, but the origins of the invading strains are known to be different. Here we tested the hypothesis, in standardized cage environments, that A. albopictus males from Brazil are less capable of satyrizing A. aegypti females than A. albopictus males from the United States. Using strains of A. aegypti and A. albopictus from the United States of known susceptibility to and capacity for interspecific mating, we demonstrate that A. albopictus colonized from collections in the Brazilian cities of Rio de Janeiro and Manaus were relatively unsuccessful in inseminating virgin female A. aegypti from Key West Florida compared to A. albopictus from peninsular Florida. We suggest that the low satyrization potential of Brazilian A. albopictus males may contribute to the lack of documented competitive displacements of A. aegypti in that country.

Overexpression of Rictor has been demonstrated to result in increased mechanistic target of rapamycin C2 (mTORC2) nucleation and activity leading to tumor growth and increased invasive characteristics in glioblastoma multiforme (GBM). However, the mechanisms regulating Rictor expression in these tumors is not clearly understood. In this report, we demonstrate that Rictor is regulated at the level of mRNA translation via heat-shock transcription factor 1 (HSF1)-induced HuR activity. HuR is shown to directly bind the 3' untranslated region of the Rictor transcript and enhance translational efficiency. Moreover, we demonstrate that mTORC2/AKT signaling activates HSF1 resulting in a feed-forward cascade in which continued mTORC2 activity is able to drive Rictor expression. RNAi-mediated blockade of AKT, HSF1 or HuR is sufficient to downregulate Rictor and inhibit GBM growth and invasive characteristics in vitro and suppress xenograft growth in mice. Modulation of AKT or HSF1 activity via the ectopic expression of mutant alleles support the ability of AKT to activate HSF1 and demonstrate continued HSF1/HuR/Rictor signaling in the context of AKT knockdown. We further show that constitutive overexpression of HuR is able to maintain Rictor expression under conditions of AKT or HSF1 loss. The expression of these components is also examined in patient GBM samples and correlative associations between the relative expression of these factors support the presence of these signaling relationships in GBM. These data support a role for a feed-forward loop mechanism by which mTORC2 activity stimulates Rictor translational efficiency via an AKT/HSF1/HuR signaling cascade resulting in enhanced mTORC2 activity in these tumors.

Phosgene is classified as a chemical warfare agent, yet data on its short-duration high concentration toxicity in a nose-only exposure rat model is sparse and inconsistent. Hence, an exposure system for short term/high concentration exposure was developed and characterized. Herein, we report the median lethal concentration (LC(50)) for a 10-min nasal exposure of phosgene in a 24-h rat survival model. Male Wistar rats (Envigo) weighing 180-210 g on the day of exposure, were exposed to phosgene gas via nose-only inhalation using a system specifically designed to allow the simultaneous exposure and quantification of phosgene. After 24 h, the surviving rats were euthanized, the lung/body mass ratio determined, and lung tissues analyzed for histopathology. Increased terminal airway edema in the lungs located primarily at the alveoli (resulting in an increased lung/body mass ratio) coincided with the observed mortality. An LC(50) value of 129.2 mg/m(3) for a 10-min exposure was determined. Furthermore, in agreement with other highly toxic compounds, this study reveals a LC(50) concentration value supportive of a non-linear toxic load model, where the toxic load exponent is > 1 (n(e) = 1.17). Thus, in line with other chemical warfare agents, phosgene toxicity is predicted to be more severe with short-duration, high-concentration exposures than long-duration, low-concentration exposures. This model is anticipated to be refined and developed to screen novel therapeutics against relevant short-term high concentration phosgene exposures expected from a terrorist attack, battlefield deployment, or industrial accident.

Interplanetary flux ropes (IFRs) observed in the solar wind have been investigated through application of the Lundquist model, which is a cylindrical flux-rope model with a constant- $\alpha $ force-free magnetic-field model. This study evaluated two Lundquist-model fitting methods by applying them to magnetic-obstacle (MO) events observed by the Wind and Solar TErrestrial RElations Observatory (STEREO) spacecraft and by comparing the results. In one method, the pitch angle of the magnetic field at the IFR boundary is assumed to be $90^{\circ}$ , whereas in the other method this restriction is relaxed and the pitch angle is handled as a free parameter [ $\alpha _{\mathrm{p}} $ ]. We found that the angle between the axial and radial directions in radial tangential normal (RTN) coordinates (cone angle) and the magnetic flux of the IFR were significantly different for approximately 30% of these events. However, both methods yielded similar values for the direction of the IFR axis projected onto the T–N plane in the RTN coordinates (tilt angle). We also found that the statistical distribution of $\alpha _{\mathrm{p}}$ , which was estimated using the generalized method, shows a spread of $34^{\circ}$ centered at $82^{\circ}$ , implying that a highly twisted magnetic-field line surrounds the surface of the IFR for approximately 60% of the events. On the other hand, it was noted that a significant number of events (approximately 25%) have a small $\alpha _{\mathrm{p}}$ ( ${

Several thousand tracers from a 2D model of a thermonuclear supernova were used in a Monte Carlo post-processing approach to determine p-nuclide abundance uncertainties originating from nuclear physics uncertainties in the reaction rates.

A series of rigid cyanuryl polyimines, polyguanamines (PGs) bearing polycyclic aromatic hydrocarbon pendants were successfully synthesized from 2-substituted 4,6-dichloro-1,3,5-triazine and aromatic diamine monomers used in conventional solution polymerization. In addition, their thermal and optical properties were also investigated. All polymers showed high thermostabilities (Tg ∼ 320 °C, Td5 (N2) ∼ 466 °C, residue at 800 °C under nitrogen ∼69.0%) and adequate solubilities in polar organic solvents. Films prepared by the solvent-cast method showed good transparencies, which mainly depended on the diamine structure as opposed to the dichloride moiety. The refractive indices at the D-line (589 nm) of the PG films were unexpectedly high, between 1.677 and 1.800, compared to those of common organic optical polymeric resins. The incorporated melamine moieties afforded effective dense packing with the polycyclic aromatic hydrocarbon groups filling the free spaces between rigid polymer chains, resulting in unusually high refractive indices. The PG polymer solution in N-methylpyrrolidone showed strong blue fluorescence (371–471 nm) with a quantum yield of up to 98%.

TP53 is a tumor suppressor gene that is mutated in 50% of cancers, and its function is tightly regulated by the E3 ligase, Mdm2. Both p53 and Mdm2 are localized in the cell nucleus, a site that is impervious to therapeutic regulation by most antibodies. We identified a cell-penetrating lupus monoclonal anti-DNA antibody, mAb 3E10, that targets the nucleus, and we engineered mAb 3E10 to function as an intranuclear transport system to deliver therapeutic antibodies into the nucleus as bispecific single chain Fv (scFv) fragments. Bispecific scFvs composed of 3E10 include PAb421 (3E10-PAb421) that binds p53 and restores the function of mutated p53, and 3G5 (3E10-3G5) that binds Mdm2 and prevents destruction of p53 by Mdm2. We documented the therapeutic efficacy of these bispecific scFvs separately in previous studies. In this study, we show that combination therapy with 3E10-PAb421 and 3E10-3G5 augments growth inhibition of cells with p53 mutations compared to the effect of either antibody alone. By contrast, no enhanced response was observed in cells with wild-type p53 or in cells homozygous null for p53.

Background Although early reperfusion is the most desirable intervention after ischemic myocardial insult, it may add to damage through oxidative stress. Objectives This study investigated the cardioprotective effects of a single intravenous dose of heat shock protein-72 (HSP72) coupled to a single-chain variable fragment (Fv) of monoclonal antibody 3E10 (3E10Fv) in a rabbit ischemia-reperfusion model. The Fv facilitates rapid transport of HSP72 into cells, even with intact membranes. Methods A left coronary artery occlusion (40 min) reperfusion (3 h) model was used in 31 rabbits. Of these, 12 rabbits received the fusion protein (Fv-HSP72) intravenously. The remaining 19 control rabbits received a molar equivalent of 3E10Fv alone (n = 6), HSP72 alone (n = 6), or phosphate-buffered saline (n = 7). Serial echocardiographic examinations were performed to assess left ventricular function before and after reperfusion. Micro–single-photon emission computed tomography imaging of 99mTc-labeled annexin-V was performed with micro–computed tomography scanning to characterize apoptotic damage in vivo, followed by gamma counting of the excised myocardial specimens to quantify cell death. Histopathological characterization of the myocardial tissue and sequential cardiac troponin I measurements were also undertaken. Results Myocardial annexin-V uptake was 43% lower in the area at risk (p = 0.0003) in Fv-HSP72–treated rabbits compared with control animals receiving HSP72 or 3E10Fv alone. During reperfusion, troponin I release was 42% lower and the echocardiographic left ventricular ejection fraction 27% higher in the Fv-HSP72–treated group compared with control animals. Histopathological analyses confirmed penetration of 3E10Fv-containing molecules into cardiomyocytes in vivo, and treatment with Fv-HSP72 showed fewer apoptotic nuclei compared with control rabbits. Conclusions Single-dose administration of Fv-HSP72 fusion protein at the time of reperfusion reduced myocardial apoptosis by almost one-half and improved left ventricular functional recovery after myocardial ischemia-reperfusion injury in rabbits. It might have potential to serve as an adjunct to early reperfusion in the management of myocardial infarction.

Insects with complex life cycles show a variety of developmental strategies when faced with low nutrient conditions requiring trade-offs in timing and nutrient storage. Previously documented strategies among insects include plasticity or canalization (fixation) of the pupation threshold, postthreshold developmental time, and adult size or stored resources. Using four models previously developed by Juliano et al. (2004), we tested the plasticity of these traits in the aquatic larvae of the frog-biting midge, Corethrella appendiculata Grabham. Rates of prey consumption and timing of pupation of fourth instar midges were compared across a range of prey densities. Model comparisons revealed that final instar C. appendiculata larvae exhibit a canalized pupation threshold followed by a canalized lengthy postthreshold period. Males entered the final instar earlier and reached a significantly lower threshold while females entered later and averaged more prey consumed daily, suggesting differing developmental strategies between sexes. Plasticity in body and egg clutch sizes of females from differing nutrient treatments was observed, but all females produced eggs autogenously. The canalized developmental pattern and long postthreshold period displayed in C. appendiculata is hypothesized to be related to the midge's preference for relatively permanent aquatic habitats.

The purpose of this paper is to prove the reliability of artificial hip joints’ surface generated by the electrolytically treated grinding process; the evaluation was carried out in a hip simulator and animal tests. The evaluation indicated that the fabricated hyper-hemispherical shaped artificial hip joints presented a high quality smoother surface than 10 nm in Ra. The fabricated surface also showed its ability to prevent crystalline phase transition during the hip simulation test, suggesting that the stably modified layer was generated via grinding process. The results of animal test showed that the fabricated surface could have a highly-improved biocompatibility.

The spread of Aedes albopictus (Skuse) eastward in the mid-1980s from its initial establishment in Houston, TX, was associated with rapid declines and local disappearances of Aedes aegypti (L.) in Gulf Coast states and Florida where annual larval surveillance during the early 1990s described temporal and spatial patterns of competitive displacements in cemeteries and tire shops. Approximately 20 yr later in 2013-2014, we re-visited former collection sites and sampled aquatic immatures of these two species from tire shops in 10 cities on State Route 441 and from 9 cemeteries from Lakeland to Miami in southwest Florida. In the recent samples Ae. aegypti was recovered from three central Florida cities where it had not been detected in 1994, but its northern limit on Rte. 441, Apopka, did not change. Other evidence, such as trends at a few cemeteries, suggested a moderate resurgence of this species since 1994. Cage experiments that exposed female progeny of Ae. aegypti from recent Florida collection sites to interspecific mating by Ae. albopictus males showed that females from coexistence sites had evolved resistance to cross-mating, but Ae. aegypti from sites with no Ae. albopictus were relatively susceptible to satyrization. Habitat classifications of collection sites were reduced by principal component (PC) analysis to four variables that accounted for > 99% of variances; PCs with strong positive loadings for tree cover and ground vegetation were associated with collection sites yielding only Ae. albopictus Within the coexistence range of the two species, the numbers of Ae. aegypti among total Aedes collected were strongly correlated in stepwise logistic regression models with two habitat-derived PCs, distance from the coast, and annual rainfall and mean maximum temperatures at the nearest weather station. Subtle increases in the range of Ae. aegypti since its previous displacements are interpreted in the context of the evolution of resistance to mating interference, realized versus fundamental niches of the two species, and persisting competition with Ae. albopictus.

The success of an invasive species in a new region depends on its interactions with ecologically similar resident species. Invasions by disease vector mosquitoes are important as they may have ecological and epidemiological consequences. Potential interactions of a recent invasive mosquito, Aedes japonicus Theobald, with resident species in Virginia were evaluated by sampling larvae from containers and trapping adults. Distinct species compositions were observed for artificial containers and rock pools, with Ae. albopictus most abundant in the former and Ae. japonicus in the latter. However, these two species were found to co-occur in 21.2% of containers sampled. Among the six mosquito species most common in containers from May through September, 2006, only interspecific associations of Ae. japonicus with Aedes albopictus (Skuse) and Aedes triseriatus (Say) were significant, and both were negative. In addition to differences in habitat preference, mean crowding estimates suggest that interspecific repulsion may contribute to the significant negative associations observed between these species. High relative abundances of late instars and pupae of Ae. japonicus seem to provide this species with a mechanism of evading competition with Ae. albopictus, facilitating their coexistence in artificial containers. Although annual fluctuations were observed, trends in adult populations over a 6-yr period provide no evidence of declines. In summary, this survey of diverse container types and all life stages provided only limited evidence for competitive displacements or reductions of resident container species by Ae. japonicus, as observed elsewhere in its invasive range.

The therapeutic use of antibodies is restricted by the limited access of antibodies to intracellular compartments. To overcome this limitation, we developed a cell-penetrating monoclonal antibody, mAb 3E10, as an intracellular delivery vehicle for the intracellular and intranuclear delivery of antibodies constructed as bispecific single-chain Fv fragments. Because MDM2 is an important target in cancer therapy, we selected monoclonal antibody (mAb) 3G5 for intracellular transport. mAb 3G5 binds MDM2 and blocks binding of MDM2 to p53. Here, we show that the resulting 3E10-3G5 bispecific antibody retains cell-penetrating and MDM2-binding activity, increases tumor p53 levels, and inhibits growth of MDM2-addicted tumors. The use of cell-penetrating bispecific antibodies in targeted molecular therapy will significantly broaden the spectrum of accessible intracellular targets and may have a profound impact in cancer therapy. Mol Cancer Ther; 11(10); 2169–73. ©2012 AACR.

Upon mating, male mosquitoes transfer accessory gland proteins (Acps) that induce refractoriness to further mating in females. This can also occur because of cross-insemination by males of related species, a process known as mating interference (satyrization). This mechanism could explain the competitive displacement of resident Aedes aegypti by the invasive Aedes albopictus where they co-occur. We tested this hypothesis in mosquito populations in Florida. A new polymerase chain reaction species diagnostic applied to sperm dissected from 304 field-collected females revealed bidirectional cross-mating in five (1.6%) individuals. Cross-injections of females with Acps showed that Ae. albopictus males induced monogamy in heterospecific females but not Ae. aegypti males. Despite its low frequency in the areas under study, the first evidence of cross-mating in nature and the asymmetric effect of Acps on mating suggest that satyrization may have initially contributed to the observed competitive reduction of Ae. aegypti by invasive Ae. albopictus in many areas.

Foxp3 is a nuclear transcription factor that is both a tumor suppressor factor and regulator of T-regulatory cell (Treg) function, and is a potential therapeutic target in both autoimmunity and cancer. In order to distinguish molecular pathways responsible for these separate Foxp3 functions, deletion mutants of Foxp3 proteins were transduced and analyzed for cytotoxic activity in human cancer cell lines Skov3, MDA-MB-231, MCF-7 and Jurkat. Human Foxp3 cDNA mutants were amplified and ligated to produce plasmids for direct cell transfection. Constructs were produced and confirmed by DNA sequencing. Lipofectamine 2000 was used for plasmid transfection. Foxp3 cells were then examined. The results of our experiments reveal retention of tumor suppressor function in the absence of NFAT binding and transcriptional activation required for Treg function. Our results have significant implications for the design of autoimmune and cancer therapies that target Foxp3 and Treg cells.

Within 2 yr of the arrival of the invasive container mosquito Aedes albopictus (Skuse), the previously dominant invasive mosquito Aedes aegypti (L.) disappeared from many Florida cemeteries. At some cemeteries, however, Ae. aegypti populations seem stable despite Ae. albopictus invasion. We sought to understand this variation in the outcome (exclusion, coexistence) of this invasion, given that previous experiments show that Ae. albopictus is the superior larval competitor. We tested experimentally the hypothesis that climate-dependent egg survivorship differs between exclusion and coexistence cemeteries and that differences in invasion outcome are associated with microclimate. Viability of eggs oviposited in the laboratory and suspended in vases at six cemeteries was significantly greater for Ae. aegypti than for Ae. albopictus, and greater in 2001 than in 2006. Cemeteries differed significantly in egg survivorship of Ae. albopictus, but not of Ae. aegypti, which is consistent with the hypothesis that Ae. albopictus suffers site-specific, climate-driven egg mortality that mitigates the competitive superiority of larval Ae. albopictus. Principal component (PC) analysis of microclimate records from vases during the experiments yielded three PCs accounting for >96% of the variance in both years of experiments. Multivariate analysis of variance of the three PCs revealed significant microclimate differences among the six cemeteries and between exclusion versus coexistence cemeteries. Stepwise logistic regression of egg survivorship versus microclimate PCs yielded significant fits for both species, and twice as much variance explained for Ae. albopictus as for Ae. aegypti in both years, Higher mortalities in 2006 were associated with high average daily maximum temperatures in vases, with lethal thresholds for both species at ≈40°C. From 1990 to 2007, vase occupancy by Ae. albopictus increased and that by Ae. aegypti decreased, with increasing seasonal precipitation at one well-sampled cemetery. Results support the hypothesis that locally variable climate-driven mortality of Ae. albopictus eggs contributes to patterns of exclusion of, or coexistence with, Ae. aegypti.