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Supplementary Figure S2. Genetic tools used for dissecting Il6/Stat3/Myc signaling. (A) Scheme of conditional alleles and PCR primers used for assessing the recombination of Pten/ Trp53 genes. (B) Senescence-specific ß-gal staining is detected after Pten loss but not after Pten/Trp53 co-deletion. Scale Bar, 200 μm.

Supplementary Figure S1. Copy Number Alteration analysis of human prostate cancer confirms loss of PTEN and TP53 genes as a genetic hallmark of metastatic prostate cancer. (A) Co-deletion frequency for PTEN and TP53 in metastatic and primary prostate cancer samples. (B) Co-deletion frequency for PTEN and TP53 in metastatic and primary prostate cancer samples. (C) Co-deletion frequency for PTEN and TP53 in metastatic prostate cancer samples . (D) Gene maps of the conditional alleles used for ablation of Pten/ Trp53 genes and activation of tdTomato. Cre leads to recombination of loxP sites in the Pten/Trp53 genes and excision of a STOP cassette that leads to activation of the fluorescence protein tdTomato. (E) Analysis of tdTomato-positive cells using Guava flow cytometry system show that AdCre leads to activation of tdTomato in over 95% percent of cells without the need for selection. (F) Activation of tdTomato can be easily visualized using fluorescence microscopy.

Supplementary Figure S7. Pathway to Pten/ Trp53 deficient prostate metastasis and therapy resistance. Pten-loss triggers PIP3 signaling to activate AKT, which prompts activation of the p53/ p21/ p16 tumor suppressors and senescence arrest. Suppression and loss of p53 in this context results in cell proliferation and Il6 secretion. Il6 signals both auto and paracrine to activate Myc via the Jak/Stat pathway. Myc induces the Phlpp2 phosphatase which creates a negative feedback loop by dampening Akt activation. Metastases (and castration-resistant tumors) select for increased Myc expression (and gene amplification, and Akt inactivation, demonstrating that Myc supersedes the need for Akt in lethal prostate cancer.

Supplementary Figure S3. Tools used for targeting of Il6 and Stat3. (A) Il6 neutralizing antibody specifically decreases the proliferation of Pten/ Trp53 deficient primary MEFs. ANOVA, Dunnett's post-hoc test, **p

Supplementary Figure S4. Activation of Stat3/Myc signaling is specific to Ptenpc-/-; Trp53pc-/-prostate and leads to stromal expansion. (A) H&E analysis of prostates from all studied genotypes reveals massive stromal expansion after co-deletion of Pten and Trp53. IHC analysis confirms stromal pStat3/ Myc activation specifically in Ptenpc-/-; Trp53pc-/- prostate. Note that pAkt activation is restricted to gland, consistent with triggering paracrine Il6/Stat3/Myc activation. Scale Bars, 100 μm. (B) Pten is present in the stroma of Ptenpc-/-; Trp53pc-/- prostates confirming that the expansion of stroma is not driven by spurious Pten/Trp53 recombination. (C) Proliferation in the Ptenpc-/- glands correlates with Akt activation, in contrast to Ptenpc-/-; Trp53pc-/- glands, where it correlates with Stat3/ Myc staining.

Supplementary Figure S6. Ar expression in primary and metastatic tumors. (A) IHC analysis of the Ar status in RapidCaP lesions shows no significant Ar staining (left panel) while most prostate glands show strong nuclear Ar staining. (B) The wild type and mutant prostates from Probasin-Cre animals show strong Ar staining.

Supplementary Figure Legends from MYC Drives Pten/Trp53-Deficient Proliferation and Metastasis due to IL6 Secretion and AKT Suppression via PHLPP2

The hindgut epithelial barrier plays an important role in maintaining absorption and immune homeostasis in ruminants. However, little information is available on changes in colon epithelial barrier structure and function following grain-induced subacute ruminal acidosis (SARA). The objective of this study was to investigate the effects of grain-induced SARA on colon epithelial morphological structure, permeability, and gene expression involved in epithelial barrier function. Twelve mid-lactating (136 ± 2 d in milk; milk yield = 1.68 ± 0.15 kg/d) Saanen dairy goats with 62.13 ± 4.76 kg of body weight were randomly divided into either the control (CON) treatment (n = 6) or SARA treatment (n = 6). The CON goats were fed a basal diet with a nonfiber carbohydrates to neutral detergent fiber ratio of 1.15 for 60 d. The SARA goats were fed 4 diets with increasing nonfiber carbohydrates to neutral detergent fiber ratio at 1.15, 1.49, 2.12, and 2.66 to induce SARA, with each diet (referred to as period) being fed for 15 d, including 12 d for adaptation and 3 d for sampling. Continuous ruminal pH recordings were used to diagnose the severity of SARA. Additionally, colonic tissues were collected to evaluate the epithelial morphological structure, permeability, and expression of tight junction proteins using transmission electron microscopy, Ussing chamber, quantitative real-time PCR, and Western blotting. Profound disruption in the colonic epithelium was mainly manifested as the electron density of tight junctions decreased, intercellular space widened, and mitochondria swelled in SARA goats. Colon epithelial short-circuit current, tissue conductance, and the mucosal-to-serosal flux of fluorescein isothiocyanate-dextran 4 kDa were increased and potential difference was decreased in SARA goats compared with CON goats. Subacute ruminal acidosis increased mRNA and protein expression levels of CLDN1 and OCLN in the colonic epithelium. Overall, the data of the present study demonstrate that SARA can impair the barrier function of the colonic epithelium at both structural and functional levels, which is associated with severe epithelial structural damage and increased permeability and changes in the expression of tight junction proteins.

A vital requirement for bulk-metallic glasses (BMGs) as structural materials is the attainment of both strength and toughness, yet invariably, as in most materials, these properties are mutually exclusive. However, by utilizing a hydrogen microalloying technology, involving alloying with a gas mixture of hydrogen/argon, we have converted “strong-yet-brittle” bulk-metallic glasses into “stronger-and-tough” ones. We combine experiments with molecular dynamics simulations to systematically analyze the atomic-scale details on how trace hydrogen additions can induce internal changes in the amorphous structure of Zr-Cu-based glassy alloys, with the aim of discerning the structural origin of the combined high strength, ductility and toughness of these materials. Our results, from both relaxation spectrum analysis and calculations of the atomic configurations, indicate that minor additions of hydrogen, instead of causing embrittlement, can have a positive influence on the mechanical properties of BMGs. Specifically, they generate more highly activated “soft spots” to promote multiple shear bands to enhance deformability, yet at the same time engender the formation of a more strengthened structural matrix to delay their initiation, a factor which can serve to elevate strength. Accordingly, our H-alloyed samples display a larger yield strength and fracture strain than the H-free ones. The current findings not only show how the strength-toughness trade-off can be successfully overcome in bulk-metallic glasses, but also we regard this understanding as a step forward to decoding the salient underlying mechanisms for the correlating structure, relaxation behavior and mechanical properties of these materials.

Physiological male sterility induced by the chemical hybridizing agent (CHA) overcomes problems of maintenance of sterile lines and restorers. However, the mechanism of sterility is unclear. The process of tapetum of CHA-treated ‘Xi’nong 2611’ at uninucleate, binucleate and trinucleate were compared with control to determine if tapetum varying differently during developmental stages. Tapetal degradation in CHA-treated ‘Xi’nong 2611’ began at late uninucleate stage, somewhat earlier than control plants. Cytological observations indicated that the gradual degradation of the tapetum in CHA-treated ‘Xi’nong 2611’ was initiated and terminated earlier than in the control. These findings implied that CHA-induced male sterility was related to abnormally early tapetal degradation. In order to indicate the role of the SKP1 gene in fertility/sterility in wheat, its expression was assessed in anthers at uninucleate, binucleate and trinucleate stages. SKP1 expression was reduced in the later developmental stages, and there was an obvious decrease from the uninucleate to trinucleate stages. Higher expression of the SKP1 gene occurred in ‘Xi’nong 2611’ compared to CHA-treated ‘Xi’nong 2611’. This implied that SKP1 gene expression was inhibited during the fertility transformation process and was related to transformation from fertility to sterility. Moreover, the results from this study suggest that SKP1 plays an essential role of conducting fertility in physiological male sterility.

Background Deletion of the chromatin remodeler chromodomain helicase DNA-binding protein 1 (CHD1) is a common genomic alteration found in human prostate cancers (PCas). CHD1 loss represents a distinct PCa subtype characterized by SPOP mutation and higher genomic instability. However, the role of CHD1 in PCa development in vivo and its clinical utility remain unclear. Patients and methods To study the role of CHD1 in PCa development and its loss in clinical management, we generated a genetically engineered mouse model with prostate-specific deletion of murine Chd1 as well as isogenic CHD1 wild-type and homozygous deleted human benign and PCa lines. We also developed patient-derived organoid cultures and screened patients with metastatic PCa for CHD1 loss. Results We demonstrate that CHD1 loss sensitizes cells to DNA damage and causes a synthetic lethal response to DNA damaging therapy in vitro, in vivo, ex vivo, in patient-derived organoid cultures and in a patient with metastatic PCa. Mechanistically, CHD1 regulates 53BP1 stability and CHD1 loss leads to decreased error-free homologous recombination (HR) repair, which is compensated by increased error-prone non-homologous end joining (NHEJ) repair for DNA double-strand break (DSB) repair. Conclusions Our study provides the first in vivo and in patient evidence supporting the role of CHD1 in DSB repair and in response to DNA damaging therapy. We uncover mechanistic insights that CHD1 modulates the choice between HR and NHEJ DSB repair and suggest that CHD1 loss may contribute to the genomic instability seen in this subset of PCas.

By analyzing large quantities of discharges in the unfavorable ion $ \vec B\times \nabla B $ drift direction, the I-mode operation has been confirmed in EAST tokamak. During the L-mode to I-mode transition, the energy confinement has a prominent improvement by the formation of a high-temperature edge pedestal, while the particle confinement remains almost identical to that in the L-mode. Similar with the I-mode observation on other devices, the $ E_r $ profiles obtained by the eight-channel Doppler backscattering system (DBS8)\cite{J.Q.Hu} show a deeper edge $ E_r $ well in the I-mode than that in the L-mode. And a weak coherent mode (WCM) with the frequency range of 40-150 kHz is observed at the edge plasma with the radial extend of about 2-3 cm. WCM could be observed in both density fluctuation and radial electric field fluctuation, and the bicoherence analyses showed significant couplings between WCM and high frequency turbulence, implying that the $ E_r $ fluctuation and the caused flow shear from WCM should play an important role during I-mode. In addition, a low-frequency oscillation with a frequency range of 5-10 kHz is always accompanied with WCM, where GAM intensity is decreased or disappeared. Many evidences show that the a low-frequency oscillation may be a novel kind of limited cycle oscillation but further investigations are needed to explain the new properties such as the harmonics and obvious magnetical perturbations., Comment: 14 pages, 7 figures

A disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS-4) can effectively degrade articular cartilage matrix proteoglycan and damage the intervertebral disc of spinal tuberculosis patients, resulting in deterioration of the physical properties of articular cartilage. Transforming growth factor β activated kinase 1 (TAK1) is similar to vascular cell adhesion molecule 1 (VCAM-1) and closely related to a variety of pathophysiological processes. This study intended to explore the expression of ADAMTS-4, VCAM-1, and TAK1 in cartilage tissue obtained from spinal tuberculosis patients and their inter-relationships, aiming to provide new treatment approaches for spinal tuberculosis. Patients with spinal tuberculosis (N = 60) from the department of orthopedics and patients with traumatic spinal fracture (N = 60, controls) were recruited for the study. ADAMTS-4, VCAM-1, and TAK1 expression was detected by immunohistochemistry. SPSS 19.0 software was used for data processing and analysis. The score values of ADAMTS-4, TAK1, and VCAM-1 were 1.45 ± 0.10, 1.33 ± 0.09, and 1.54 ± 0.11, respectively, which were significantly higher than those in normal controls (P < 0.05). ADAMTS-4 showed positive correlation with VCAM-1 and TAK1. ADAMTS-4, TAK1, and VCAM-1 expressions increased in spinal tuberculosis patients. They could provide clinical reference for spinal tuberculosis diagnosis and new treatment strategies can be devised by focusing on their positive correlation.

Underground excavations such as tunnels, storage caverns, and underground power plants have been constructed throughout the world. Rock masses in underground constructions always contain discontinuities such as faults, joints, bedding planes, and fractures. These discontinuities intersect with each other to create rock blocks [1]. One of the most important problems in underground excavation is the accidental falling of rock blocks at the working faces [2]. To predict this, the removability and stability of the rock blocks around the underground tunnels and caverns must be evaluated based on the characteristics of the discontinuities in the rock masses. The most commonly used method is the block theory developed by Goodman and Shi [3]. A complete block theory analysis consists of removable block identification based on geometric and topological methods, mode analysis to determine whether the removable block has a mode of failure or not, and stability evaluation to identify the key blocks by incorporating comparatively simple mechanics analysis. After that, the supporting force and directions are designed to make sure that the key blocks are stable. If the key blocks are stable, the entire rock mass will be stable [4]. Block theory has been applied widely in rock engineering since early 1990s [5]. However, its stability evaluation is based on static analysis. Recent studies demonstrate that sometimes an external dynamic disturbance may dominate the system behavior. The traditional stability analysis based on static analysis may underestimate the rock instability if not taking these dynamic factors into account. Earlier researchers [6–8] observed that when a pulse loading is applied to a blocky rock system, friction between the blocks in the orthogonal direction to the pulse loading may be significantly reduced or even disappear, which is known as an ultra low friction

A platinum overlayer was prepared on Au nanoparticles that were electrodeposited on a graphite electrode by replacement of the under-potential deposition (UPD) Cu monolayer. The micrographs and element composition of Au nanoparticles and UPD Cu monolayers were characterized by scanning electron microscopy (SEM) and energy disperse X-ray spectroscopy (EDS), respectively. The electrocatalytic properties of Pt overlayer/Au/graphite electrode for methanol oxidation have been investigated by cyclic voltammetry (CV). High electroactivity and good long-term stability can be observed. These results indicate that the core (Au nanoparticle)-shell (Pt overlayer) nanocatalyst obtained by the above method may have good potential applications in designing noble metal catalysts with super low loading and high activity at the atomic level in fuel cells.

A novel sample treatment, namely vortex-assisted dispersive liquid-liquid microextraction (VDLLME), followed by GC/MS has been developed for the determination of gaseous phase polycyclic aromatic hydrocarbons (PAHs) emitted from a marine diesel engine. The method integrates extraction, enrichment, and concentration into one step by combining sample collection and VDLLME into a much shorter and accurate procedure. We have optimized several important variables influencing the extraction efficiency including type and amount of extraction solvent, type and volume of disperser solvent, sample pH, salting-out effect, vortex and centrifugation time. Under the optimum conditions, high enrichment factors (more than 1,000) and acceptable total spiked recoveries (72.0-95.1%) were obtained using GC/MS detection. The limits of detection and quantification of the proposed method were in the ranges of 0.1-0.5 ng/L. Our total pretreatment can be competed in less than 0.5 hours so as to decreases loss of PAHs by 18.37% compared to traditional methods.

The monomer triethyleneglycoldimethacrylate (TEGDMA) is used as a diluent in many resin-based dental materials. It was previously shown in vitro that TEGDMA was released into the adjacent biophase from such materials during the first days after placement. In this study, the uptake, distribution, and excretion of 14C-TEGDMA applied via gastric, intradermal, and intravenous administration at dose levels well above those encountered in dental care were examined in vivo in guinea pigs and mice as a test of the hypothesis that TEGDMA reaches cytotoxic levels in mammalian tissues. 14C-TEGDMA was taken up rapidly from the stomach and small intestine after gastric administration in both species and was widely distributed in the body following administration by each route. Most 14C was excreted within one day as 14 CO2. The peak equivalent TEGDMA levels in all mouse and guinea pig tissues examined were at least 1000-fold less than known toxic levels. The study therefore did not support the hypothesis.

The primary objective of this study is to perform a blinded evaluation of two groups of retrospective image registration techniques, using as a gold standard a prospective marker-based registration method, and to compare the performance of one group with the other. These techniques have already been evaluated individually [27]. In this paper, however, we find that by grouping the techniques as volume based or surface based, we can make some interesting conclusions which were not visible in the earlier study. In order to ensure blindness, all retrospective registrations were performed by participants who had no knowledge of the gold-standard results until after their results had been submitted. Image volumes of three modalities: X-ray computed tomography (CT), magnetic resonance (MR), and positron emission tomography (PET) were obtained from patients undergoing neurosurgery at Vanderbilt University Medical Center on whom bone-implanted fiducial markers were mounted. These volumes had all traces of the markers removed and were provided via the Internet to project collaborators outside Vanderbilt, who then performed retrospective registrations on the volumes, calculating transformations from CT to MR and/or from PET to MR. These investigators communicated their transformations, again via the Internet, to Vanderbilt, where the accuracy of each registration was evaluated. In this evaluation, the accuracy is measured at multiple volumes of interest (VOI's). Our results indicate that the volume-based techniques in this study tended to give substantially more accurate and reliable results than the surface-based ones for the CT-to-MR registration tasks, and slightly more accurate results for the PET-to-MR tasks. Analysis of these results revealed that the rotational component of error was more pronounced for the surface-based group. It was also apparent that all of the registration techniques we examined have the potential to produce satisfactory results much of the time, but that visual inspection is necessary to guard against large errors.

A compact neural network algorithm for partial-response maximum-likelihood (PRML) sequence detection is presented. Compact neural networks are a class of locally connected neural networks suitable for very large scale integration (VLSI) implementation. The hardware complexity for VLSI implementation of the proposed algorithm grows linearly with the level of the deliberately designed symbol interference effects of the partial-response (PR) signalling scheme. Large dedicated memory for storage of likelihood matrices in digital Viterbi-algorithm-based detectors is not needed for the proposed detector. Detailed analysis on network stability for network topology and time constant of an analog neuron is described. This detector algorithm has competitive bit-error rate performance when compared with the digital Viterbi algorithm under the noise condition for many real-world applications. The proposed algorithm is suitable for analog VLSI implementation because of its low time complexity and linear area complexity for the detection of PRML signalling schemes.

Very large scale integration (VLSI) compact neural network architecture for maximum-likelihood detector of partial response (PR) communication receivers is presented. The compact neural network approach has many attractive advantages in achieving low power, low cost, compact chip area, and faster processing speed by its loosely coupled parallel processing nature. In this paper, the design of a state-constrained analog neural processor, and the corresponding parallel architecture to realize the PR detection algorithms and the related scalability and performance evaluation issues are described with detailed design analysis. A design example of a PR IV detector has been used to demonstrate the advantages of such a scalable massive VLSI architecture. A processing rate of 265 Mb/s was achieved with SPICE simulation for a prototype PR IV detector on a silicon area of 5.14 mm/spl times/5.81 mm in a 1.2 /spl mu/m CMOS technology. An estimated processing capacity of 886 Mb/s can be achieved if the same design is scaled up to a 1.0 cm/sup 2/ silicon area for the same technology. Such promising performance potential clearly indicates that VLSI compact neural network detector can meet the needs in future high speed data communication systems at very low cost.

An image processing technique is presented for finding and localizing the centroids of cylindrical markers externally attached to the human head in computed tomography (CT) and magnetic resonance (MR) image volumes. The centroids can be used as control points for image registration. The technique, which is fast, automatic, and knowledge-based, has two major steps. First, it searches the entire image volume to find one voxel inside each marker-like object. The authors call this voxel a "candidate" voxel, and they call the object a candidate marker. Second, it classifies the voxels in a region surrounding the candidate voxel as marker or nonmarker voxels using knowledge-based rules and calculates an intensity-weighted centroid for each true marker. The authors call this final centroid the "fiducial" point of the marker. The technique was developed on 42 scans of six patients-one CT and six MR scans per patient. There are four markers attached to each patient for a total of 168 marker images. For the CT images the false marker rate was zero. For MR the false marker rate was 1.4% (Two out of 144 markers). To evaluate the accuracy of the fiducial points, CT-MR registration was performed after correcting the MR images for geometrical distortion. The fiducial registration accuracy averaged 0.4 mm and was better than 0.6 mm for each of the eighteen image pairs.

The behavior of non-thermal plasma (NTP) and combined plasma catalysis (CPC) was investigated for removal of low-concentration benzene, toluene and p -xylene (BTX mixture) in air using a link tooth wheel-cylinder plasma reactor. Combining NTP with MnO x /Al 2 O 3 catalyst after the discharge zone (CPC) significantly promoted BTX conversion and improved the energy efficiency. For a specific input energy (SIE) of 10 J L −1 , the conversion of benzene, toluene and p -xylene reached 94%, 97% and 95%, respectively. The introduction of MnO x /Al 2 O 3 catalyst also moved the BTX conversion towards total oxidation and reduced the emission of O 3 and NO 2 as compared to NTP alone. For an SIE of 10 J L −1 , the O 3 outlet concentration decreased from 46.7 for NTP alone to 1.9 ppm for CPC, while the NO 2 emission correspondingly decreased from 1380 to 40 ppb.

1. Intracellular recordings were made from antidromically identified motoneurones in transverse spinal cord slices from neonatal (12-16 day) rats. 2. Superfusion of (+/-)-baclofen (0.5-50 microM) reduced the excitatory postsynaptic potentials (e.p.s.ps) and inhibitory postsynaptic potentials (i.p.s.ps) evoked by dorsal root or dorsal root entry zone stimulation in a concentration-dependent manner; the calculated EC50 was 2.4 microM. Baclofen in comparable concentrations also reversibly eliminated spontaneously occurring e.p.s.ps and i.p.s.ps. 3. (-)-Baclofen was more effective as compared to baclofen in reducing the synaptic responses, whereas (+)-baclofen at concentrations as high as 50 microM was ineffective. 4. Baclofen (less than 5 microM) attenuated the synaptic responses without causing a significant change of passive membrane properties and depolarizations induced by exogenously applied glutamate. In addition to synaptic depression, baclofen (greater than 5 microM) caused a hyperpolarization associated with decreased membrane resistance in some of the motoneurones; the glutamate responses were also attenuated. 5. Baclofen reversibly depressed the spike after-hyperpolarization of the motoneurones. 6. GABA (1-10 mM) depressed synaptic transmission and depolarized or hyperpolarized motoneurones. While potentiated by the uptake inhibitor nipecotic acid, the synaptic depressant effect of GABA was not antagonized by bicuculline. 7. The synaptic depressant effect of baclofen was neither blocked by GABAA antagonists bicuculline and picrotoxin (10-50 microM) nor by the GABAB antagonist phaclofen (0.1-1 mM). 8. It is suggested that baclofen depresses excitatory and inhibitory transmission in rat motoneurones by primarily a presynaptic mechanism in reducing the liberation of chemical transmitters from nerve endings via a phaclofen-insensitive GABAB receptor.

Describes an extrinsic-point-based, interactive image-guided neurosurgical system designed at Vanderbilt University, Nashville, TN, as part of a collaborative effort among the Departments of Neurological Surgery, Computer Science, and Biomedical Engineering. Multimodal image-to-image (II) and image-to-physical (IP) registration is accomplished using implantable markers. Physical space tracking is accomplished with optical triangulation. The authors investigate the theoretical accuracy of point-based registration using numerical simulations, the experimental accuracy of their system using data obtained with a phantom, and the clinical accuracy of their system using data acquired in a prospective clinical trial by 6 neurosurgeons at 4 medical centers from 158 patients undergoing craniotomies to respect cerebral lesions. The authors can determine the position of their markers with an error of approximately 0.4 mm in X-ray computed tomography (CT) and magnetic resonance (MR) images and 0.3 mm in physical space. The theoretical registration error using 4 such markers distributed around the head in a configuration that is clinically practical is approximately 0.5-0.6 mm. The mean CT-physical registration error for the: phantom experiments is 0.5 mm and for the clinical data obtained with rigid head fixation during scanning is 0.7 mm. The mean CT-MR registration error for the clinical data obtained without rigid head fixation during scanning is 1.4 mm, which is the highest mean error that the authors observed. These theoretical and experimental findings indicate that this system is an accurate navigational aid that can provide real-time feedback to the surgeon about anatomical structures encountered in the surgical field.