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Detection of tumor targeting by in vivo imaging (fluorescent intensity, FLI)of CEA TCB formats having bivalent or monovalent binding to CEA and directly labeled with Alexa647.

Pharmacokinetic profiles of CEA TCB in mice, additional information on CD3 signaling, T cell activation, tumor cell lysis and cytokine release upon CEA TCB and untargeted control TCB treatment.

Purpose: CEA TCB is a novel IgG-based T-cell bispecific (TCB) antibody for the treatment of CEA-expressing solid tumors currently in phase I clinical trials (NCT02324257). Its format incorporates bivalent binding to CEA, a head-to-tail fusion of CEA- and CD3e-binding Fab domains and an engineered Fc region with completely abolished binding to FcγRs and C1q. The study provides novel mechanistic insights into the activity and mode of action of CEA TCB.Experimental Design: CEA TCB activity was characterized on 110 cell lines in vitro and in xenograft tumor models in vivo using NOG mice engrafted with human peripheral blood mononuclear cells.Results: Simultaneous binding of CEA TCB to tumor and T cells leads to formation of immunologic synapses, T-cell activation, secretion of cytotoxic granules, and tumor cell lysis. CEA TCB activity strongly correlates with CEA expression, with higher potency observed in highly CEA-expressing tumor cells and a threshold of approximately 10,000 CEA-binding sites/cell, which allows distinguishing between high- and low-CEA–expressing tumor and primary epithelial cells, respectively. Genetic factors do not affect CEA TCB activity confirming that CEA expression level is the strongest predictor of CEA TCB activity. In vivo, CEA TCB induces regression of CEA-expressing xenograft tumors with variable amounts of immune cell infiltrate, leads to increased frequency of activated T cells, and converts PD-L1 negative into PD-L1–positive tumors.Conclusions: CEA TCB is a novel generation TCB displaying potent antitumor activity; it is efficacious in poorly infiltrated tumors where it increases T-cell infiltration and generates a highly inflamed tumor microenvironment. Clin Cancer Res; 22(13); 3286–97. ©2016 AACR.

Representative videos of intravital two-photon (2P) microscopy of tumors treated with CEA TCB. Tumor cells red, T cells green.

Representative videos of intravital two-photon (2P) microscopy of tumors treated with untargeted TCB. Tumor cells red, T cells green.

List, Source and CEA binding sites of Cell Lines used in the study

Assessment of binding of CEA TCB to human and cynomolgus monkey CEA- and CD3-expressing cells, internalization and ADCC, CDC activity.

Supplemental Material and Methods describing CEA TCB production and purification process, determination of affinity and avidity, genetic characterization of cell lines, determination of CEA binding sites, assessment of T cell proliferation and activation, Jurkat NFAT assay, confocal microscopy, FRET, SDPK, in vivo efficacy and imaging studies. Supplemental Video Legends are also within the file.

PURPOSE The aim of this study was to evaluate the diagnostic performance of 18F-choline PET and MRI in patients with primary hyperparathyroidism. Furthermore, the additional value of software-based PET/MRI scan fusion was analyzed. PATIENTS AND METHODS This retrospective study includes 42 patients (38 women) with an age between 32.5 and 79.1 years. PET/CT scans were performed on a dedicated system after injection of 250 to 350 MBq 18F-choline. For the MRI examination, T1-weighted images of the cervical region were used. The image fusion was made by anatomical coregistration using an automated algorithm based on mutual information. RESULTS A total of 46 lesions were discovered and histologically confirmed in 42 patients. Histopathological examination revealed 38 adenomas and 8 hyperplasias. This means that, in 4 of these 42 patients, 2 lesions per patient were discovered. PET/CT also detected 46 abnormal findings, but only 43 were correctly recognized, whereas the other 3 were false-positive (FP). Six lesions could not be detected correctly: 3 were FP and 3 false-negative, which resulted in a sensitivity of 93.5% and a specificity of 97.5%. The site-specific evaluation showed 18 true-positive enlarged parathyroid glands with MRI, but also produced 13 FP findings and failed to detect 28 lesions; the sensitivity and specificity are thus 39.1% and 89.3%, respectively. The difference in detection rate between 18F-choline PET/CT and MRI was statistically significant (P < 0.001). CONCLUSIONS 18F-choline PET/CT is clearly superior to MRI for localization diagnostics in primary hyperparathyroidism. Image fusion of both modalities can be helpful for more precise anatomical assignment.

Resilient Watershed Management differs from traditional approaches in that it incorporates climate change and natural disaster risk into the overall watershed management planning process. Underpinning these two attributes is its use of a landscape approach to natural resource management and risk mitigation, which factors in all aspects of land use and societal needs, from the highest snow mountains, to the middle hills, to the coastal plains. Most importantly, the umbrella under which all three concepts fall is the sustainable development of a watershed’s inhabitants. Resilient Watershed Management also incorporates an integration of the physical and social sciences as fundamental to its operations, from the first analysis of stakeholder vulnerabilities to the baseline data required for the most meaningful monitoring and evaluation plan. The use of local knowledge within the planning process is also stressed. Case studies from FAO projects in Morocco and Pakistan provide evidence of the benefits of this approach. In both cases, the investment and objectives addressed land degradation while engaging the local communities and income generation. In Morocco, the project’s objective was combating soil erosion and reducing the risk of flooding in overgrazed and eroded area. The investments included the improvement of irrigation systems, apple orchards, promoting income-generation activities, and enhancing the capacity of local institutions. In Pakistan, landslide stabilization was conducted through integrated and participatory watershed management. Activities included bioengineering and soil conservation measures, capacity development of stakeholders, strengthening natural resource management-based enterprises, and the improvement of local livelihoods. In both cases, the involvement of governmental institutions was key in ensuring the follow-up and replication of the projects in similarly affected areas. It is through adaptive management, the integration of key stakeholders and institutions, and the recognition of the landscape as the unit of action, that these communities successfully enhanced their risk management.

The world’s forests provide fundamental protection of soil and water resources as well as multiple ecosystem services and cultural or spiritual values. We summarized the FRA 2015 data for protective functions and ecosystem services, and analyzed increasing or decreasing trends of protective areas. The global forest area managed for protection of soil and water was 1.002billionha as of 2015, which was 25.1% of all global forested areas. Protective forests have increased by 0.181billionha over the past 25years mainly because more countries are now reporting protective forest areas (139 in 2015 vs 114 in 1990). However, average percentage of designated for protective forests did not change significantly from 1990 to 2015. Global forest area managed for ecosystem services is also now at 25.4% of global total forest area and has changed little over the past 25years. Among the twelve categories of protective forests, flood control, public recreation, and cultural services increased both in terms of percentage of total forest area and the number of reporting countries. Public awareness of the importance of forest resources for functions and services other than production continues to increase as evidenced by the increase of protective forest designations and reporting in many countries. Percentages of total forest area designated for both protective forests and ecosystem services show a dual-peak distribution of numbers of countries concentrated at 0% and 100%. This suggests a socio-economic influence for the designations. We examined five case study countries (Australia, Canada, China, Kenya, and Russia). The most dramatic changes in the past 25years have been in China where protective forests for soil and water resources increased from about 12% to 28% of forest area. The Russian Federation has also increased percentages of forest area devoted to soil and water resource protection and delivery of ecosystem services. Australia is now reporting in more protective forest categories whereas Kenya and Canada changed little. These five countries have their own classification of forest functions and recalculation methods of reporting for FRA 2015 were different. This demonstrates the difficulty in establishing a universal common designation scheme for multi-functions of forest. Production of more accurate assessments by further improvements in the reporting framework and data quality would help advance the value of FRA as the unique global database for forest functions integrated between forest ecosystems and social sciences.

As many of the world’s poor and food-insecure people live in mountain regions, sustainable mountain development is an important part of the work of the Food and Agriculture Organization (FAO) of the United Nations (UN). Over the years, FAO has played the leading role in sustainable mountain development within the UN system; it was appointed task manager for chapter 13 of agenda 21 in 1992 and acted as the lead agency for the International Year of Mountains in 2002. From 2003 onward, FAO was also mandated by the UN General Assembly to lead the annual observance of International Mountain Day on 11 December. Every 2 years, FAO prepares the secretary general’s report to the UN General Assembly, which describes the status and progress of sustainable mountain development at the national and international levels and provides suggestions for consideration by the Assembly. FAO is a member of the Mountain Partnership and hosts its secretariat.This review of the latest actions of FAO’s program on sustainable mountain development, watershed management, and forest hydrology—which includes normative work, field activities, and support to international processes—updates our previous statement (Manuelli et al, 2014) and summarizes the latest achievements of the Mountain Partnership.

Eponyms are used almost daily in the clinical practice of dermatology. And yet, information about the person behind the eponyms is difficult to find. Indeed, who is? What is this person’s nationality? Is this person alive or dead? How can one find the paper in which this person first described the disease? Eponyms are used to describe not only disease, but also clinical signs, surgical procedures, staining techniques, pharmacological formulations, and even pieces of equipment. In this article we present the symptoms starting with (M) and other. The symptoms and their synonyms, and those who have described this symptom or phenomenon.

With its mandate to work on natural resource management, food security, and livelihoods and its attention to the social, economic, and environmental dimensions of sustainable development, the Food and Agriculture Organization of the United Nations (FAO) has played a leading role in sustainable mountain development for many years. In 1992, FAO was appointed task manager for Chapter 13 of Agenda 21 entitled Managing Fragile Ecosystems: Sustainable Mountain Development (UNCED 1992) and acted as the lead agency for the International Year of Mountains in 2002. FAO is a member of the Mountain Partnership and hosts its Secretariat. From 2003 onward, FAO has also been mandated by the United Nations General Assembly to lead the annual observance of International Mountain Day on 11 December. Every two years, FAO prepares the Secretary General's Report to the United Nations General Assembly, in which it describes the status of sustainable mountain development at the national and international levels and pro...

In the future Smart Grid, numerous stakeholders within the electric power grid need to exchange information in order to realize applications like customer energy feedback, billing and invoicing of variable tariffs, demand side management and efficient charging of electric vehicles. Using a Service-oriented Architecture (SOA) based on Web services promises a convenient way to provide a data infrastructure capable to realize the required interactions in a flexible way. A main concern in such an architecture is how access to data can be controlled in order to prevent security or privacy violations. Access control depends strongly on authentication and authorization mechanisms. Therefore, this paper contributes i) a SOA for the Smart Grid and ii) an access control mechanism that is taken into consideration from the early beginning of the system design. Furthermore, a proof of concept implementation and a scalability analysis are used to investigate the requirements on computational resources in a large scale deployment.

U nepovoljnim geotehničkim uvjetima objekti se ne mogu uspješno temeljiti metodama iz uobičajene inženjerske prakse. U uvjetima neheterogenog tla najveći problem predstavlja procjena veličine ukupnih deformacija i nejednakog slijeganja. Primjenom tzv. naprednih konstitucijskih modela tla moguće je analitički pouzdano procijeniti međusobnu interakciju temeljnog tla i inženjerske strukture. Napredni geotehnički modeli poput Hardening soil modela zahtijevaju poznavanje niza parametara kojima se opisuje ovisnost modula stišljivosti tla o stanju napona, povijesti nanošenja opterećenja i stanju konsolidacije. Rezultat analize je pouzdano određivanje modula reakcije tla ispod temelja u svrhu nastavka proračuna u nekom od programa za statičku analizu konstrukcija. Učinkovita metoda temeljenja u nepovoljnim geotehničkim uvjetima je kombinirano temeljenje temeljnom pločom i pilotima.

We have previously described an approach whereby antibody Fc fragments harboring a single C-terminal selenocysteine residue (Fc-Sec) are directed against a variety of targets by changing the peptide or small molecule to which they are conjugated. In the present work, we describe methodology for improving the efficacy of these Fc-Sec conjugates by incorporating cytotoxic drugs. The Fc-Sec protein is first programmed to target specific tumor cell types by attachment of a bifunctional linker that contains a "clickable" handle (e.g., cyclobutane or cyclooctyne) in addition to a tumor cell-binding peptide or small molecule. Following Fc-Sec conjugation, a cytotoxic warhead is then attached by cycloaddition reactions of tetrazine or azide-containing linker. To validate this approach, we used a model system in which folic acid (FA) is the targeting moiety and a disulfide-linked biotin moiety serves as a cytotoxic drug surrogate. We demonstrated successful targeting of Fc-Sec proteins to folate-receptor expressing tumor cells. Tetrazine ligation was found to be an efficient method for biotin "arming" of the folate-targeted Fc-Sec proteins. We also report novel bioconjugation methodologies that use [4 + 2] cycloaddition reactions between tetrazines and cyclooctynes.

Mountain ecosystems and watersheds are essential for long-term sustainable global development and poverty alleviation and can make an important contribution to climate change adaptation and mitigation. Freshwater, rich biodiversity, and other natural resources provided by mountains are vital for the livelihood of billions of people. However, recent environmental, economic, and social developments such as climate change, increasing natural disasters, population growth, the expansion of commercial agriculture, and urbanization compromise the ability of mountain ecosystems and watersheds to provide essential environmental goods and services. Degradation and decreasing water flows seriously affect agricultural production and food security and threaten the supply of water to large urban centers in the lowlands, while water, energy, and food are likely to be the main scarcities in the coming decades. Watershed management and sustainable mountain development (SMD) are necessary and appropriate approache...

With its mandate to work on natural resource management; food security; and livelihoods; and its attention to the social, economic, and environmental dimensions of sustainable development, the Food and Agriculture Organization of the United Nations (FAO) has played a leading role in sustainable mountain development for many years. In 1992, the FAO was appointed Task Manager for Chapter 13 of Agenda 21 (Managing Fragile Ecosystems: Sustainable Mountain Development) and acted as the lead agency for the International Year of Mountains in 2002. The FAO hosts the global Secretariat of the Mountain Partnership and, from 2003 onward, has also been mandated by the United Nations General Assembly to lead observance of the International Mountain Day, every year on 11 December. Over time, the FAO has progressively built up a conceptual and operational framework that links sustainable mountain development to forest hydrology and watershed and risk management. This Platform Statement provides an update on the...

Die Behandlung von Wunden ist heute zunehmend die Domäne von speziell aus- und weitergebildeten medizinischen Fachkräften. Die Flut von unterschiedlichen Wundpflegemassnahmen und Verbandtechniken wächst kontinuierlich. Studien vergleichen deren Wirksamkeit und selbst die Medizinalperson droht in der Vielfalt von Resultaten den Halt im Bewährten zu verlieren. Die vorgestellte Kasuistik zeigt, dass auch eine nichtmedizinische Massnahme, nämlich ein Wechsel der behandelnden Personen – mit anderen Worten zwischenmenschliche Aspekte – wesentlich zur Wundheilung beitragen kann.

Purpose: Gene expression profiling identified receptor tyrosine kinase ROR1, an embryonic protein involved in organogenesis, as a signature gene in B-cell chronic lymphocytic leukemia (B-CLL). To assess the suitability of ROR1 as a cell surface antigen for targeted therapy of B-CLL, we carried out a comprehensive analysis of ROR1 protein expression. Experimental Design: Peripheral blood mononuclear cells, sera, and other adult tissues from B-CLL patients and healthy donors were analyzed qualitatively and quantitatively for ROR1 protein expression by flow cytometry, cell surface biotinylation, Western blotting, and ELISA. Results: ROR1 protein is selectively expressed on the surface of B-CLL cells, whereas normal B cells, other normal blood cells, and normal adult tissues do not express cell surface ROR1. Moreover, cell surface expression of ROR1 is uniform and constitutive, i.e., independent of anatomic niches, independent of biological and clinical heterogeneity of B-CLL, independent of B-cell activation, and found at similar levels in all B-CLL samples tested. The antibody binding capacity of B-CLL cell surface ROR1 was determined to be in the range of 103 to 104 molecules per cell. A portion of B-CLL cell surface ROR1 was actively internalized upon antibody binding. Soluble ROR1 protein was detectable in sera of Conclusions: The restricted, uniform, and constitutive cell surface expression of ROR1 protein in B-CLL provides a strong incentive for the development of targeted therapeutics such as monoclonal antibodies.

NgR1, NgR2, and NgR3 which constitute the Nogo-66 receptor family are primarily expressed by neurons in the central nervous system (CNS) and believed to limit axonal growth and sprouting following CNS injury. In an attempt to define the expression and decipher the function of individual members of the Nogo-66 receptor family, we previously reported the generation of selective rabbit polyclonal antibodies. Here we exploit the same immune repertoires by phage display technology to generate rabbit monoclonal antibodies (mAbs) with nanomolar affinity to epitopes that are specific for NgR1 and NgR2, respectively, but at the same time conserved between mouse, rat, and human orthologs. Employing phage display vector pC3C, a newly designed phagemid optimized for the generation and selection of Fab libraries with human constant domains, rabbit mAbs were selected from chimeric rabbit/human Fab libraries, characterized in terms of specificity, affinity, and amino acid sequence, and finally converted to chimeric rabbit/human IgG. Using immunofluorescence microscopy and immunoprecipitation, we demonstrate strong and specific recognition of cell surface bound Nogo-66 receptor family members by chimeric rabbit/human IgG. The rabbit mAbs reported here together with their amino acid sequences constitute a defined panel of species cross-reactive reagents in infinite supply which will aid investigations toward a functional role of the Nogo-66 receptor family in and beyond the CNS.