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Figure S1. CD133 expression correlated with PSA expression. Figure S2. CD133 expression in xenografts. Figure S3. Characterization of labeled HA10. Table S1. Ex vivo biodistribution data.

Fig. S1. Plasmid map of A.TSTAPEG101KB in pGL3- Basic Fig. S2. PEG10 staining in positive control and healthy tissue Fig. S3. TSTA Schematic Fig. S4. Luciferase expression in empty pGL3 vector Fig. S5. PEG10 expression by IHC in HT-29 xenografts Fig. S6. HSV-TK and PEG10 expression in CWR-R1 tumors Table S1. Primers for PEG10 promoter cloning.

Purpose:Aggressive variant prostate cancer (AVPC) is a nonandrogen receptor–driven form of disease that arises in men in whom standard-of-care therapies have failed. Therapeutic options for AVPC are limited, and the development of novel therapeutics is significantly hindered by the inability to accurately quantify patient response to therapy by imaging. Imaging modalities that accurately and sensitively detect the bone and visceral metastases associated with AVPC do not exist.Experimental Design:This study investigated the transmembrane protein CD133 as a targetable cell surface antigen in AVPC. We evaluated the expression of CD133 by microarray and IHC analysis. The imaging potential of the CD133-targeted IgG (HA10 IgG) was evaluated in preclinical prostate cancer models using two different imaging modalities: near-infrared and PET imaging.Results:Evaluation of the patient data demonstrated that CD133 is overexpressed in a specific phenotype of AVPC that is androgen receptor indifferent and neuroendocrine differentiated. In addition, HA10 IgG was selective for CD133-expressing tumors in all preclinical imaging studies. PET imaging with [89Zr]Zr-HA10 IgG revealed a mean %ID/g of 24.30 ± 3.19 in CD133-positive metastatic lesions as compared with 11.82 ± 0.57 in CD133-negative lesions after 72 hours (P = 0.0069). Ex vivo biodistribution showed similar trends as signals were increased by nearly 3-fold in CD133-positive tumors (P < 0.0001).Conclusions:To our knowledge, this is the first study to define CD133 as a targetable marker of AVPC. Similarly, we have developed a novel imaging agent, which is selective for CD133-expressing tumors, resulting in a noninvasive PET imaging approach to more effectively detect and monitor AVPC.

The retrotransposon-derived paternally expressed gene 10 (PEG10) protein is ordinarily expressed at high levels in the placenta. Recently, it was discovered that PEG10 isoforms promote the progression of prostate cancer to a highly lethal androgen receptor (AR)-negative phenotype. The presence of PEG10 in other subtypes of prostate cancer has not been explored and a utility for PEG10 overexpression has not been developed. Here, we found that in addition to AR-null disease, PEG10 was also expressed in prostate cancer with constitutively active AR-splice variants. A molecular genetic imaging strategy for noninvasive imaging of AR-splice variant prostate cancer was developed by utilizing the cancer specificity of the PEG10 promoter to drive the expression of reporter genes. Plasmid insertion of a PEG10 promoter sequence optimized for enhanced output upstream of a reporter gene allowed detection of prostate cancer by near-infrared and positron emission tomography imaging after systemic administration of the plasmid in vivo. PEG10 expressing subcutaneous xenograft and intratibial tumor models were imaged by both modalities using this molecular genetic imaging strategy. This study demonstrates a preclinical proof-of-concept that the PEG10 promoter is a powerful and specific tool that can be utilized for noninvasive detection of aggressive prostate cancer subtypes.Significance:PEG10 is expressed by prostate cancer with constitutively active AR-splice variants that can be exploited for noninvasive molecular imaging of this aggressive prostate cancer subytpe.

Automated intelligent decision-making systems, working with the use of mathematical methods of data processing, can reduce the influence of the human factor in the analysis, reduce the time spent on research, improve the accuracy and reliability of the results. They help automate the quality control process and enable association material properties to its microstructure. Numerical and experimental studies of the material microstructure can be implemented using hybrid methods with the introduction of computer simulation methods. The paper proposes to use the OpenCV technology for image analysis of cast iron with spherical graphite inclusions, followed by classification of the recognized material. The optimal parameters of image preprocessing and edge recognition are set. The result of the presented work is the distribution function of inclusions depending on their concentration on the plane.

The study of the material microstructure allows obtaining information about the state of the part without additional tests and full-scale experiments. The paper offers computer methods for constructing parametric, statistically equivalent models of cast iron microstructure with the inclusion of spheroidal graphite. The studied material has a transient microstructure that exhibits variability at various material points. To analyze the unsteadiness of deformations, the Monte Carlo method is used. A finite element model is constructed to find the elastic characteristics of the material. The stress state is considered based on plane models. Numerical experiments are carried out for various concentrations of inclusions. The results obtained for the elastic constants are statistically averaged, and the dependences of the Poisson’s ratio, the moduli of elasticity, and the shear moduli on the concentration of the inclusions are established. For veracity assessment, the values obtained are compared with those obtained using the mixture rule. The results of the application of the rule confirm the correctness of the built models. The yield surfaces are found, going beyond the surface indicates the appearance of plastic strains in the material.

The retrotransposon-derived paternally expressed gene 10 (PEG10) protein is ordinarily expressed at high levels in the placenta. Recently, it was discovered that PEG10 isoforms promote the progression of prostate cancer to a highly lethal androgen receptor (AR)-negative phenotype. The presence of PEG10 in other subtypes of prostate cancer has not been explored and a utility for PEG10 overexpression has not been developed. Here, we found that in addition to AR-null disease, PEG10 was also expressed in prostate cancer with constitutively active AR-splice variants. A molecular genetic imaging strategy for noninvasive imaging of AR-splice variant prostate cancer was developed by utilizing the cancer specificity of the PEG10 promoter to drive the expression of reporter genes. Plasmid insertion of a PEG10 promoter sequence optimized for enhanced output upstream of a reporter gene allowed detection of prostate cancer by near-infrared and positron emission tomography imaging after systemic administration of the plasmid in vivo. PEG10 expressing subcutaneous xenograft and intratibial tumor models were imaged by both modalities using this molecular genetic imaging strategy. This study demonstrates a preclinical proof-of-concept that the PEG10 promoter is a powerful and specific tool that can be utilized for noninvasive detection of aggressive prostate cancer subtypes. Significance: PEG10 is expressed by prostate cancer with constitutively active AR-splice variants that can be exploited for noninvasive molecular imaging of this aggressive prostate cancer subytpe.

The modern world is often faced with the problem of the equipment design lifetime ending. Researches related to determines the residual lifetime, extension of service life, and prediction of failure-free operation is important, especially when it comes to equipment for nuclear and thermal power plants. Such interest is associated with the difficult economic situation, high cost of equipment and its components, requirements for safe working conditions, etc. The main objective of this work is to study the probability of the centrifugal pump failure-free operation. Attention is paying to the water elbow part of a centrifugal pump. Finding the probability of model failures is based on data-driven yield surface application. Takes into account the different behavior of composite materials under tensile and compressive loads, which is analyzed at the micro-level using the finite element method. Going beyond the yield surface indicates the possibility of transition into a plastic state. The proposed method of analysis of a centrifugal pump type WD 16/25 leads to predict the probability of failure during normal operation and in hydro testing mode. Consideration of the influence of corrosion-erosive processes and uniform thinning of the water pump elbow wall, an analysis of the probability of failure-free operation in time is carried out.

With long-term operation of pumping and other equipment, there is often a need to extend the life and assess the residual life. Modern life-time assessment methods are based on three-dimensional modelling and the finite element method. Within the framework of these methods, geometric models are constructed that take into account the thinning of the walls of the pumps as a result of operation. By constructing geometric models, finite element models are constructed. These models are made taking into account the conditions of loading of the structure, tightening of bolted connections, technological and temperature loads. The constructed models allow to assess the strength of the pump casing and the main bolted connections during normal operation and gyro testing. The paper assesses the cyclic strength of these structural elements. The results of the work confirmed a sufficient residual life-time for safety pump operation in the next 15 years.

The Gaching high-sulfidation epithermal deposit in the Maletoyvayam ore field features a wide range of Se-containing minerals and selenides, as well as complex gold oxides, Au tellurides (calaverite, krennerite) and native gold typical for epithermal deposits. Pyrite included in quartzites and quartz-alunite rocks was probably formed during an early stage of the ore-forming process. During the following Au-rich stage, the $f_{{\rm S}{\rm e}_{\rm 2}}$/$f_{{\rm S}_{\rm 2}}$ increased with $f_{{\rm O}_{\rm 2}}$ being relatively high, resulting in the formation of very rare compounds that have not been previously described in nature. These include Au2Te4(Se,S)3, Se3Te2, AuSe and Au(Te,Se,S) phases. The Au2Te4(Se,S)3 compounds have some variations in composition: the complete isomorphic series between Au2Te4Se3 and Au2Te4S3 was observed. The gold and Au-minerals at the main ore stage can be stable within a range of log$f_{{\rm O}_{\rm 2}}$ of −27.3 and atmospheric oxygen (?); log$f_{{\rm S}{\rm e}_{\rm 2}}$ between −12.4 and −5.7; log$f_{{\rm T}{\rm e}_{\rm 2}}$ between −10.5 and −7.8; and log$f_{{\rm S}_{\rm 2}}$ between −12.8 and −6.8 (at 250°C). The increasing oxygen fugacity during the final stage of mineralization resulted in the formation of complex Sb,As,Te,S-bearing Au oxides. Gold-oxide formation occurs due to oxidation of Au-tellurides. The final products of this process are newly-formed secondary mustard gold and Te–Se solid solutions.

With long-term operation of pumping and other equipment, there is often a need to extend the life and assess the residual life. Modern life-time assessment methods are based on three-dimensional modelling and the finite element method. Within the framework of these methods, geometric models are constructed that take into account the thinning of the walls of the pumps as a result of operation. By constructing geometric models, finite element models are constructed. These models are made taking into account the conditions of loading of the structure, tightening of bolted connections, technological and temperature loads. The constructed models allow to assess the strength of the pump casing and the main bolted connections during normal operation and gyro testing. The paper assesses the cyclic strength of these structural elements. The results of the work confirmed a sufficient residual life-time for safety pump operation in the next 15 years.

Purpose: Aggressive variant prostate cancer (AVPC) is a nonandrogen receptor–driven form of disease that arises in men in whom standard-of-care therapies have failed. Therapeutic options for AVPC are limited, and the development of novel therapeutics is significantly hindered by the inability to accurately quantify patient response to therapy by imaging. Imaging modalities that accurately and sensitively detect the bone and visceral metastases associated with AVPC do not exist. Experimental Design: This study investigated the transmembrane protein CD133 as a targetable cell surface antigen in AVPC. We evaluated the expression of CD133 by microarray and IHC analysis. The imaging potential of the CD133-targeted IgG (HA10 IgG) was evaluated in preclinical prostate cancer models using two different imaging modalities: near-infrared and PET imaging. Results: Evaluation of the patient data demonstrated that CD133 is overexpressed in a specific phenotype of AVPC that is androgen receptor indifferent and neuroendocrine differentiated. In addition, HA10 IgG was selective for CD133-expressing tumors in all preclinical imaging studies. PET imaging with [89Zr]Zr-HA10 IgG revealed a mean %ID/g of 24.30 ± 3.19 in CD133-positive metastatic lesions as compared with 11.82 ± 0.57 in CD133-negative lesions after 72 hours (P = 0.0069). Ex vivo biodistribution showed similar trends as signals were increased by nearly 3-fold in CD133-positive tumors (P < 0.0001). Conclusions: To our knowledge, this is the first study to define CD133 as a targetable marker of AVPC. Similarly, we have developed a novel imaging agent, which is selective for CD133-expressing tumors, resulting in a noninvasive PET imaging approach to more effectively detect and monitor AVPC.

We present new geochemical and mineralogical data from the Southern-2 orebody, located in the South-Western branch of the Talnakh intrusion (Russia) and composed of massive and disseminated sulphides. The Southern-2 orebody consists dominantly of chalcopyrite along with lesser pentlandite and subordinate cubanite, bornite and pyrrhotite, and differs from Cu-rich ores of the Oktyabr'sky deposit due to the absence of moikhukite-talnakhite assemblages. Ores from the Southern-2 orebody are highly enriched in copper, nickel and platinum-group elements (PGEs): Cu up to 28.12 and 10.58 wt%, Ni up to 6.29 and 2.45 wt%, and Pt + Pd up to 220 and 173 ppm in the massive and disseminated ores respectively. The ratios of Cu/Ni = 3.85–7.34 in massive and 2.64–6.58 in disseminated ores. The ratios of the platinum to iridium group elements (PPGE/IPGE) reaches more than 40000. These features indicate an extremely different degree of fractionation of the sulphide melt during the formation of ores. The sequence of crystallization of the PGE minerals (PGMs) on the basis of textural information was as follows: Pt-Pd-Sn (rustenburgite, atokite) → Pd-Cu-Sn (stannopalladinite) → Pt-Fe-Cu-Ni (tetraferroplatinum) → Pd-Ni-As (majakite) → Pd-As (palladoarsenide) → Pd-Pb(Bi) (zvyagintsevite, polarite and Pd5Pb3 unnamed phase) → Au-Cu-Pd (tetra-auricupride and auricupride) → Au-Ag alloys. This is consistent with experimental data on PGE minerals. The PGM assemblage was caused by the evolution of a significantly fractionated sulphide melt that was enriched in Cu, Ni, Pt, Pd, Pb and Au, and as a result produced minerals with peculiar compositions (e.g., Cu-rich stannopalladinite, Pb-rich polarite, Ni-rich palladoarsenide), which do not occur in other ores. Furthermore, Pt-Fe-Cu-Ni alloys are abundant and sperrylite is absent, which is a very common phase in all ores of the Noril'sk area. The composition of pentlandite with Ni/(Ni + Fe) ratios ranging from 0.48 to 0.68 indicates that it formed between 560 °C (Ni-poor) to 500 °C (Ni-rich) and −10.5 to −7 lgfS2, respectively. The PGMs and Au-minerals were formed at temperatures ranging from 550 to 225 °C. The specificity of sulphide (with PGMs) association is due to the evolution of the Cu-rich (and Ni-rich) melt under elevated sulphur fugacity conditions. The mechanism of filter-pressing, which precipitated sulphide drops at the bottom of the chamber, and penetration of sulphide melts into the country rocks is supported by the same geochemical composition of disseminated and massive ores.

The recent success of autologous T cell-based therapies in hematological malignancies has spurred interest in applying similar immunotherapy strategies to the treatment of solid tumors. Identified nearly 4 decades ago, natural killer (NK) cells represent an arguably better cell type for immunotherapy development. Natural killer cells are cytotoxic lymphocytes that mediate the direct killing of transformed cells with reduced or absent major histocompatibility complex (MHC) and are the effector cells in antibody-dependent cell-mediated cytotoxicity. Unlike T cells, they do not require human leukocyte antigen (HLA) matching allowing for the adoptive transfer of allogeneic NK cells in the clinic. The development of NK cell-based therapies for solid tumors is complicated by the presence of an immunosuppressive tumor microenvironment that can potentially disarm NK cells rendering them inactive. The molecular imaging of NK cells in vivo will be crucial for the development of new therapies allowing for the immediate assessment of therapeutic response and off-target effects. A number of groups have investigated methods for detecting NK cells by optical, nuclear, and magnetic resonance imaging. In this review, we will provide an overview of the advances made in imaging NK cells in both preclinical and clinical studies.

The M1 family of metalloproteases represents a large number of exopeptidases that cleave single amino acid residues from the N-terminus of peptide substrates. One member of this family that has been well studied is aminopeptidase N (APN), a multifunctional protease known to cleave biologically active peptides and aide in coronavirus entry. The proteolytic activity of APN promotes cancer angiogenesis and metastasis making it an important target for cancer therapy. To understand the substrate specificity of APN for the development of targeted inhibitors, we used a global substrate profiling method to determine the P1–P4′ amino acid preferences. The key structural features of the APN pharmacophore required for substrate recognition were elucidated by x-ray crystallography. By combining these substrate profiling and structural data, we were able to design a selective peptide inhibitor of APN that was an effective therapeutic both in vitro and in vivo against APN-expressing prostate cancer models.

// Mariya Shapovalova 1 , Julia Davydova 2 , Christine Henzler 3 , Mark Daniel 4 , Scott M. Dehm 3 , Christopher A. Warlick 5 and Aaron M. LeBeau 1 1 Department of Pharmacology, University of Minnesota, Minneapolis 55455, MN, USA 2 Department of Surgery, University of Minnesota, Minneapolis 55455, MN, USA 3 Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis 55455, MN, USA 4 Department of Microbiology and Immunology, University of Minnesota, Minneapolis 55455, MN, USA 5 Department of Urology, University of Minnesota, Minneapolis 55455, MN, USA Correspondence to: Aaron M. LeBeau, email: alebeau@umn.edu Keywords: α-methylacyl-CoA racemase; molecular imaging; adenovirus; prostate cancer Received: October 17, 2018 Accepted: November 16, 2018 Published: November 30, 2018 ABSTRACT The metabolic protein alpha-methylacyl-CoA racemase (AMACR) is significantly overexpressed in prostate cancer compared to the normal prostate and other non-malignant tissue. Though an attractive target, there are no reports in the literature on leveraging the expression of AMACR for the molecular imaging of prostate cancer. Here, we used a molecular-genetic imaging strategy to exploit the transcriptional specificity of the AMACR promoter for the in vivo detection of prostate cancer using the reporter gene luciferase. We performed a stepwise truncation of the promoter and identified a 565 base pair minimal promoter for AMACR that retained both high activity and specificity. Following identification of the minimal promoter for AMACR, we used an advanced two-step transcriptional amplification system to maximize the promoter output. We showed that our optimized AMACR promoter can drive expression of luciferase for molecular imaging in subcutaneous xenograft models of androgen receptor-positive and androgen receptor-negative prostate cancer using a non-replicative adenovirus for gene delivery. Our results provide evidence that the AMACR promoter can be exploited to drive the cancer-specific expression of reporter genes and potentially even be incorporated into conditionally replicative adenoviruses for oncolytic therapy and other applications.

The retrotransposon-derived paternally expressed gene 10 (PEG10) protein is ordinarily expressed at high levels in the placenta. The over-expression of PEG10 has been documented in a number of malignancies including hepatocellular carcinoma and leukemia. Recently, it was discovered that PEG10 isoforms promoted the progression of prostate adenocarcinoma to a highly lethal non-androgen receptor (AR) driven subtype called aggressive variant prostate cancer (AVPC). In this study, we found that PEG10 was also expressed in castration-resistant PCa possessing constitutively active AR-splice variants in addition to AR-negative AVPC. We subsequently developed a molecular genetic imaging strategy for the non-invasive imaging of PCa by utilizing the cancer specificity of the PEG10 promoter to drive the expression of reporter genes. The transcriptional output of the PEG10 promoter was enhanced to enable high expression of reporter genes by multiple imaging modalities and immunoassay. By using this PEG10 promoter upstream of a reporter gene in a plasmid, we were able to detect PCa by fluorescence and positron emission tomography (PET) imaging after systemic administration of the plasmid in mice. This method also allowed for the detection of PCa by an immunoassay of mouse urine. Our study demonstrates a pre-clinical proof-of-concept that the PEG10 promoter is a powerful and specific tool that can be utilized for non-invasive detection of highly lethal PCa subtypes. This biomarker imaging technology has the potential to improve detection and can be further modified for therapeutic applications. Citation Format: Mariya Shapovalova. Biomarker-driven molecular imaging of prostate cancer using the PEG10 promoter [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1145.