Murine leukemia virus (MLV) replication monitored with fluorescent proteins

Background: Cancer gene therapy will benefit from vectors that are able to replicate in tumor tissue and cause a bystander effect. Replication-competent murine leukemia virus (MLV) has been described to have potential as cancer therapeutics, however, MLV infection does not cause a cytopathic effect in the infected cell and viral replication can only be studied by immunostaining or measurement of reverse transcriptase activity. Results: We inserted the coding sequences for green fluorescent protein (GFP) into the proline-rich region (PRR) of the ecotropic envelope protein (Env) and were able to fluorescently label MLV. This allowed us to directly monitor viral replication and attachment to target cells by flow cytometry. We used this method to study viral replication of recombinant MLVs and split viral genomes, which were generated by replacement of the MLV env gene with the red fluorescent protein (RFP) and separately cloning GFP-Env into a retroviral vector. Co-transfection of both plasmids into target cells resulted in the generation of semi-replicative vectors, and the two color labeling allowed to determine the distribution of the individual genomes in the target cells and was indicative for the occurrence of recombination events. Conclusions: Fluorescently labeled MLVs are excellent tools for the study of factors that influence viral replication and can be used to optimize MLV-based replication-competent viruses or vectors for gene therapy

Vaccination directed against the human endogenous retrovirus-K envelope protein inhibits tumor growth in a murine model system

Human endogenous retrovirus (HERV) genomes are chromosomally integrated in all cells of an individual. They are normally transcriptionally silenced and transmitted only vertically. Enhanced expression of HERV-K accompanied by the emergence of anti-HERV-K-directed immune responses has been observed in tumor patients and HIV-infected individuals. As HERV-K is usually not expressed and immunological tolerance development is unlikely, it is an appropriate target for the development of immunotherapies. We generated a recombinant vaccinia virus (MVA-HKenv) expressing the HERV-K envelope glycoprotein (ENV), based on the modified vaccinia virus Ankara (MVA), and established an animal model to test its vaccination efficacy. Murine renal carcinoma cells (Renca) were genetically altered to express E. coli beta-galactosidase (RLZ cells) or the HERV-K ENV gene (RLZ-HKenv cells). Intravenous injection of RLZ-HKenv cells into syngenic BALB/c mice led to the formation of pulmonary metastases, which were detectable by X-gal staining. A single vaccination of tumor-bearing mice with MVA-HKenv drastically reduced the number of pulmonary RLZ-HKenv tumor nodules compared to vaccination with wild-type MVA. Prophylactic vaccination of mice with MVA-HKenv precluded the formation of RLZ-HKenv tumor nodules, whereas wild-type MVA-vaccinated animals succumbed to metastasis. Protection from tumor formation correlated with enhanced HERV-K ENV-specific killing activity of splenocytes. These data demonstrate for the first time that HERV-K ENV is a useful target for vaccine development and might offer new treatment opportunities for diverse types of cancer

Abduction in Annotated Probabilistic Temporal Logic

Annotated Probabilistic Temporal (APT) logic programs are a form of logic programs that allow users to state (or systems to automatically learn)rules of the form ``formula G becomes true K time units after formula F became true with L to U% probability.\u27\u27 In this paper, we develop a theory of abduction for APT logic programs. Specifically, given an APT logic program Pi, a set of formulas H that can be ``added\u27\u27 to Pi, and a goal G, is there a subset S of H such that Pi cup S is consistent and entails the goal G? In this paper, we study the complexity of the Basic APT Abduction Problem (BAAP). We then leverage a geometric characterization of BAAP to suggest a set of pruning strategies when solving BAAP and use these intuitions to develop a sound and complete algorithm

Murine leukemia virus (MLV) replication monitored with fluorescent proteins

BACKGROUND: Cancer gene therapy will benefit from vectors that are able to replicate in tumor tissue and cause a bystander effect. Replication-competent murine leukemia virus (MLV) has been described to have potential as cancer therapeutics, however, MLV infection does not cause a cytopathic effect in the infected cell and viral replication can only be studied by immunostaining or measurement of reverse transcriptase activity. RESULTS: We inserted the coding sequences for green fluorescent protein (GFP) into the proline-rich region (PRR) of the ecotropic envelope protein (Env) and were able to fluorescently label MLV. This allowed us to directly monitor viral replication and attachment to target cells by flow cytometry. We used this method to study viral replication of recombinant MLVs and split viral genomes, which were generated by replacement of the MLV env gene with the red fluorescent protein (RFP) and separately cloning GFP-Env into a retroviral vector. Co-transfection of both plasmids into target cells resulted in the generation of semi-replicative vectors, and the two color labeling allowed to determine the distribution of the individual genomes in the target cells and was indicative for the occurrence of recombination events. CONCLUSIONS: Fluorescently labeled MLVs are excellent tools for the study of factors that influence viral replication and can be used to optimize MLV-based replication-competent viruses or vectors for gene therapy

catena-Poly[calcium-bis­[μ-N-(dimethyl­phosphino­yl)benzene­sulfonamidato]]

The crystal structure of the title calcium complex, [Ca(C8H11NO5PS)2]n, is composed of a polymeric chain, which is formed due to two bridging sulfonyl groups linking CaII ions in a O—S—O—Ca manner. Thus, the coordination environment of the CaII ions is composed of six O atoms belonging to the phosphoryl and sulfonyl groups of two chelate rings and two additional O atoms of two bridging sulfonyl groups. The coordination polyhedron of the central atom (2 symmetry) has a distorted octa­hedral geometry

catena-Poly[neodymium(III)-bis­[μ-N-(dimorpholinophosphor­yl)benzene­sulfonamidato]-sodium(I)-bis­[μ-N-(dimorpholinophosphor­yl)benzene­sulfonamidato]]

The cubic crystal structure of the title compound, [NaNd(C14H21N3O5PS)4]n, is composed of one-dimensional polymeric chains propagating in [100], built up from [Nd(C14H21N3O5PS)4]− anions and sodium cations functioning as linkers. In the complex anion, the Nd3+ ion has an eightfold coordination environment formed by the sulfonyl and phosphoryl O atoms of four bidentate chelating N-(dimorpholinophosphor­yl)benzene­sulfonamidate ligands: the resulting NdO8 polyhedron can be described as inter­mediate between dodeca­hedral and square anti­prismatic. The sodium ion adopts an NaO4 tetra­hedral geometry arising from four monodentate benzene­sulfonamidate ligands. The resulting crystal structure is unusual because it contains substantial voids (800 Å3 per unit cell), within which there is no evidence of included solvent

2,2′-Dihydroxybiphenyl-3,3′-di­carb­aldehyde dioxime

The mol­ecule of the title compound, C14H12N2O4, lies across a crystallographic inversion centre situated at the mid-point of the C—C intra-annular bond. The mol­ecule is not planar, the dihedral angle between the aromatic rings being 50.1 (1)°. The oxime group is in an E position with respect to the –OH group and forms an intra­molecular O—H⋯N hydrogen bond. In the crystal structure, inter­molecular O—H⋯O hydrogen bonds link mol­ecules into chains propagating along [001]. The crystal structure is further stabilized by inter­molecular stacking inter­actions between the rings [centroid-to-centroid distance = 3.93 (1) Å], resulting in layers parallel to the bc plane

Optimizing University Mobility : An Internal Navigation and Crowd Management System

In the evolving landscape of educational technology, the article explores the critical frontier of indoor navigation systems, focusing on universities. Traditional approaches in higher education often fall short of meeting dynamic user expectations, necessitating revolutionary solutions. This research introduces an innovative internal navigation and crowd management system that seamlessly integrates augmented reality, natural language processing, machine learning, and image processing technologies. The Android platform serves as the foundation, harnessing augmented reality's transformative capabilities to provide real-time visual cues and personalized wayfinding experiences. The voice interaction module, backed by NLP and ML, creates an intelligent, context-aware assistant. The crowd management module, employing advanced image processing, delivers real-time crowd density insights. Personalized recommendations, powered by NLP and ML, offer tailored canteen suggestions based on user preferences. The agmented reality navigation module, using Mapbox, Unity Hub, AR Core, and Vuforia, enriches the user experience with dynamic visual cues. Results reveal the success of each module: the voice interaction module showcases continuous learning, user-centric feedback, contextual guidance excellence, robust security, and multimodal interaction flexibility. The crowd management module excels in video feed processing, image processing with OpenCV, and real-time availability information retrieval. The personalized recommendations module demonstrates high accuracy, equilibrium, and robust performance. The AR navigation module impresses with precision, enriched navigation, and tailored routes through machine learning. This cohesive system sets new benchmarks for user-centric technology in universities. Future work includes multi-university integration, intelligent spatial design, and real-time decision support, paving the way for more efficient, user-centered university experiences and contributing to the advancement of smart university environments. The research serves as a pivotal force in reshaping interactions within university spaces, envisioning a future where technology seamlessly enhances the essence of human interaction in educational environments

Diaquabis­[3-(hydroxy­imino)­butanoato]nickel(II)

In the neutral, mononuclear title complex, [Ni(C4H6NO3)2(H2O)2], the Ni atom lies on a crystallographic inversion centre within a distorted octa­hedral N2O4 environment. Two trans-disposed anions of 3-hydroxy­imino­butanoic acid occupy four equatorial sites, coordinated by the deprotonated carboxyl­ate and protonated oxime groups and forming six-membered chelate rings, while the two axial positions are occupied by the water O atoms. The O atom of the oxime group forms an intra­molecular hydrogen bond with the coordinated carboxyl­ate O atom. The complex mol­ecules are linked into chains along b by hydrogen bonds between the water O atom and the carboxyl­ate O of a neighbouring mol­ecule. The chains are linked by further hydrogen bonds into a layer structure