Development of a highly potent bispecific antibody format targeting the novel tumor-specific antigen CLDN18.2

Due to multiple immune evasion mechanisms of cancer cells, novel therapy approaches are required to overcome the limitations of existing immunotherapies. Bispecific antibodies are potent anti-cancer drugs, which redirect effector T cells for specific tumor cell lysis, thus enabling the patient’s immune system to fight cancer cells. The antibody format used in this proof of concept study–bispecific ideal monoclonal antibodies termed BiMAB–is a tailor-made recombinant protein, which consists of two fused scFv antibodies recognizing different antigens. Both are arranged in tandem on a single peptide chain and the individual variable binding domains are separated by special non-immunogenic linkers. The format is comprised of a scFv targeting CLDN18.2–a gastric cancer tumor associated antigen (TAA) –while the second specificity binds the CD3 epsilon (CD3ε) subunit of the T cell receptor (TCR) on T cells. For the first time, we compared in our IMAB362-based BiMAB setting, four different anti-CD3-scFvs, respectively derived from the mAbs TR66, CLB-T3, as well as the humanized and the murine variant of UCHT1. In addition, we investigated the impact of an N- versus a C-terminal location of the IMAB362-derived scFv and the anti-CD3-scFvs. Thus, nine CLDN18.2 specific BiMAB proteins were generated, of which all showed a remarkably high cytotoxicity towards CLDN18.2-positive tumor cells. Because of its promising effectiveness, 1BiMAB emerged as the BiMAB prototype. The selectivity of 1BiMAB for its TAA and CD3ε, with affinities in the nanomolar range, has been confirmed by in vitro assays. Its dual binding depends on the design of an N-terminally positioned IMAB362 scFv and the consecutive C-terminally positioned TR66 scFv. 1BiMAB provoked a concentration and target cell dependent T cell activation, proliferation, and upregulation of the cytolytic protein Granzyme B, as well as the consequent elimination of target cells. Our results demonstrate that 1BiMAB is able to activate T cells independent of elements that are usually involved in the T cell recognition program, like antigen presentation, MHC restriction, and co-stimulatory effector molecules. In the first in vivo studies using a subcutaneous xenogeneic tumor mouse model in immune incompetent NSG mice, we could prove a significant therapeutic effect of 1BiMAB with partial or complete tumor elimination. The initial in vitro RIBOMAB experiments correspondingly showed encouraging results. The electroporation of 1BiMAB IVT-RNA into target or effector cells was feasible, while the functionality of translated 1BiMAB was proven by induced T cell activation and target cell lysis. Accordingly, we could show that the in vitro RIBOMAB approach was applicable for all nine BiMABs, which proves the RIBOMAB concept. Thus, the CLDN18.2-BiMAB strategy offers great potential for the treatment of cancer. In the future, administered either as protein or as IVT-RNA, the BiMAB format will contribute towards finding solutions to raise and sustain tumor-specific cellular responses elicited by engaged and activated endogenous T cells. This will potentially enable us to overcome immune evasion mechanisms of tumor cells, consequently supporting current solid gastric cancer therapies.

Sensor-based evaluation of Circadian motor behavior in people with Dementia. Development and validation of analysis strategies

[ITA]La demenza consiste nel deterioramento, spesso progressivo, dello stato cognitivo di un individuo. Chi è affetto da demenza, presenta alterazioni a livello cognitivo, comportamentale e motorio, ad esempio compiendo gesti ossessivi, ripetitivi, senza uno scopo preciso. La condizione dei pazienti affetti da demenza è valutata clinicamente tramite apposite scale e le informazioni relative al comportamento vengono raccolte intervistando chi se ne occupa, come familiari, il personale infermieristico o il medico curante. Spesso queste valutazioni si rivelano inaccurate, possono essere fortemente influenzate da considerazioni soggettive, e sono dispendiose in termini di tempo. Si ha quindi l'esigenza di disporre di metodiche oggettive per valutare il comportamento motorio dei pazienti e le sue alterazioni patologiche; i sensori inerziali indossabili potrebbero costituire una valida soluzione, per questo scopo. L'obiettivo principale della presente attività di tesi è stato definire e implementare un software per una valutazione oggettiva, basata su sensori, del pattern motorio circadiano, in pazienti affetti da demenza ricoverati in un'unità di terapia a lungo termine, che potrebbe evidenziare differenze nei sintomi della malattia che interessano il comportamento motorio, come descritto in ambito clinico. Lo scopo secondario è stato quello di verificare i cambiamenti motori pre- e post-intervento in un sottogruppo di pazienti, a seguito della somministrazione di un programma sperimentale di intervento basato su esercizi fisici. --------------- [ENG]Dementia involves deterioration, often progressive, of a person's cognitive status. Those who suffer from dementia, present alterations in cognitive and motor behavior, for example performing obsessive and repetitive gestures, without a purpose. The condition of patients suffering from dementia is clinically assessed by means of specific scales and information relating to the behavior are collected by interviewing caregivers, such as the family, nurses, or the doctor. Often it turns out that these are inaccurate assessments that may be heavily influenced by subjective evaluations and are costly in terms of time. Therefore, there is the need for objective methods to assess the patients' motor behavior and the pathological changes; wearable inertial sensors may represent a viable option, so this aim. The main objective of this thesis project was to define and implement a software for a sensor-based assessment of the circadian motor pattern in patients suffering from dementia, hospitalized in a long-term care unit, which could highlight differences in the disease symptoms affecting the motor behavior, as described in the clinical setting. The secondary objective was to verify pre- and post-intervention changes in the motor patterns of a subgroup of patients, following the administration of an experimental program of intervention based on physical exercises.

Transgenerational effects of temperature on egg and larval development in Cyprinodon variegatus

Transgenerational plasticity (TGP), a type of maternal effect, occurs when the environment experienced by one or both the parents prior to fertilization directly translates, without changing DNA sequences, into changes in offspring reaction norms. Evidence of such effects has been found in several traits throughout many phyla, and, although of great potential importance - especially in a time of rapid climate change - TGP in thermal growth physiology had never been demonstrated for vertebrates until the first experiment on thermal TGP in sheepshead minnows, who, given sufficient time, adaptively program their offspring for maximal egg viability and growth at the temperature experienced before fertilization. This study on sheepshead minnows from South Carolina and Connecticut investigates how population, parent temperature, and offspring temperature affect egg production, size, viability, larval survival and growth rates, whether these effects provide evidence of TGP, and whether and how they vary with length of exposure time (5, 12, 19, 26, 33 and 43 days) of the parents to the new experimental temperatures of either 26°C or 32°C. Several results are consistent with those obtained in the previous TGP study, which outline a sequence of events consisting of an initial adjustment period to the new temperatures, in which egg production decreases and no signs of TGP are present, followed by a shift to TGP (towards 26-33 days of exposure) in which parents start to produce more eggs which are better adapted to the new thermal environment. Other results present new information, such as signs of TGP in the parent temperature effect on egg sizes already around 20 days of exposure. The innovative idea of populations being able to adapt to rapidly shifting environments through non-genetic mechanisms such as TGP opens new possibilities of survival of species and will have important implications on ecology, physiology, and contemporary evolution.

Applied Developmental Psychology as an Implementation of the Life-Span View of Human Development

Application of knowledge about psychological development should, ideally, be theory based. As such, these applications represent “natural ontogenetic experiments”; the results of the evaluation of such interventions feed back to the theory, helping to support, falsify, or refine the ideas from the theory which led to the particular application. Such applied developmental intervention research is central within a currently popular perspective of life-span human development. Thus, applied developmental intervention research provides critical tests of such key concepts within this life-span perspective as: plasticity; multidirectionality; the synthesis of continuous and discontinuous processes across ontogeny; contextual embeddedness; and the role of individuals as agents in their own development. This paper elucidates some of the major features of the dynamic linkage between applied developmental psychology and this view of life-span human development. Key elements of this life-span perspective and the facts of developmental intervention, as seen from this perspective, are specified. Finally, the doctoral training program at the authors' institution is presented as one example of how this link may be institutionalized in the form of graduate education.

State Pharmaceutical Assisstance for the Elderly and Disabled: Planning Issues for Program and Non-Program States

The Medicare Catastrophic Coverage Act (MCCA) would have mandated federal assistance for Medicare beneficiaries who have high annual prescription medication costs, High national expenditures for such drugs have encouraged the development of private and state insurance programs to help with these costs. Ten state pharmaceutical assistance programs (SPAPs), designed to help certain elderly, low income, or disabled people, exist for those ineligible for Medicaid or unable to purchase coverage privately. Coordination of state and federal benefits was a consideration for established programs, and programs being planned needed to determine the feasibity of integration of federal assistance. But the enactment and subsequent appeal of the Act affected both planning and policy implications for these SPAPs. All U.S. states and territories were surveyed before the bill's repeal to collect data on the effects of MCCA for those with prescription drug programs and those without. The repeal of the federal program places pressure on the nonprogram states to proceed, perhaps more cautiously, to initiate program; for their own residents, given increasing out-of-pocket and insurance costs, and no federal program.

Postgraduate training program

BACKGROUND: Faculties face the permanent challenge to design training programs with well-balanced educational outcomes, and to offer various organised and individual learning opportunities. AIM: To apply our original model to a postgraduate training program in rheumatology in general, and to various learning experiences in particular, in order to analyse the balance between different educational objectives. METHODS: Learning times of various educational activities were reported by the junior staff as targeted learners. The suitability of different learning experiences to achieve cognitive, affective and psychomotor learning objectives was estimated. Learning points with respect to efficacy were calculated by multiplication of the estimated learning times by the perceived appropriateness of the educational strategies. RESULTS: Out of 780 hours of professional learning per year (17.7 hours/week), 37.7% of the time was spent under individual supervision of senior staff, 24.4% in organised structured learning, 22.6% in self-studies, and 15.3% in organised patient-oriented learning. The balance between the different types of learning objectives was appropriate for the overall program, but not for each particular learning experience. Acquisition of factual knowledge and problem solving was readily aimed for during organised teaching sessions of different formats, and by personal targeted reading. Attitudes, skills and competencies, as well as behavioural and performance changes were mostly learned during caring for patients under interactive supervision by experts. CONCLUSION: We encourage other faculties to apply this approach to any other curriculum of undergraduate education, postgraduate training or continuous professional development in order to foster the development of well-balanced learning experiences.

INVESTIGATION OF STRATEGIES TO PROMOTE EFFECTIVE TEACHER PROFESSIONAL DEVELOPMENT EXPERIENCES IN EARTH SCIENCE

This dissertation serves as a call to geoscientists to share responsibility with K-12 educators for increasing Earth science literacy. When partnerships are created among K-12 educators and geoscientists, the synergy created can promote Earth science literacy in students, teachers, and the broader community. The research described here resulted in development of tools that can support effective professional development for teachers. One tool is used during the planning stages to structure a professional development program, another set of tools supports measurement of the effectiveness of a development program, and the third tool supports sustainability of professional development programs. The Michigan Teacher Excellence Program (MiTEP), a Math/Science Partnership project funded by the National Science Foundation, served as the test bed for developing and testing these tools. The first tool, the planning tool, is the Earth Science Literacy Principles (ESLP). The ESLP served as a planning tool for the two-week summer field courses as part of the MiTEP program. The ESLP, published in 2009, clearly describe what an Earth science literate person should know. The ESLP consists of nine big ideas and their supporting fundamental concepts. Using the ESLP for planning a professional development program assisted both instructors and teacher-participants focus on important concepts throughout the professional development activity. The measurement tools were developed to measure change in teachers’ Earth science content-area knowledge and perceptions related to teaching and learning that result from participating in a professional development program. The first measurement tool, the Earth System Concept Inventory (ESCI), directly measures content-area knowledge through a succession of multiple-choice questions that are aligned with the content of the professional development experience. The second measurement, an exit survey, collects qualitative data from teachers regarding their impression of the professional development. Both the ESCI and the exit survey were tested for validity and reliability. Lesson study is discussed here as a strategy for sustaining professional development in a school or a district after the end of a professional development activity. Lesson study, as described here, was offered as a formal course. Teachers engaged in lesson study worked collaboratively to design and test lessons that improve the teachers’ classroom practices. Data regarding the impact of the lesson study activity were acquired through surveys, written documents, and group interviews. The data are interpreted to indicate that the lesson study process improved teacher quality and classroom practices. In the case described here, the lesson study process was adopted by the teachers’ district and currently serves as part of the district’s work in Professional Learning Communities, resulting in ongoing professional development throughout the district.

Increasing intercultural competence and tolerance in multicultural schools: A Training Program and its effectiveness

This study reports the implementation of a Training of Intercultural Competence and Tolerance (TICT) for upper-secondary school students and the empirical evaluation of its effectiveness. The TICT program was developed to counteract increasing interethnic conflicts in the North Caucasus Federal District of Russia. It is based on the theoretical and empirical framework of social psychology and cross-cultural psychology. The training effectiveness was assessed by conducting pre- and post-surveys among the training participants. The results indicate that TICT contributes to the development of a positive ethnic identity and the formation of a civic identity among the participating youth. It also increases their optimism regarding the future of interethnic relations in Russia and the subjective level of intercultural competence of majority group youth towards minority cultures. Thus, the evaluation of the training effectiveness of the TICT has shown that the aims of the training have been achieved to a large extent and that the Training of Intercultural Competence and Tolerance can be effectively used to prevent interethnic conflicts and promote interethnic relations in multicultural schools. Suggestions for the practical implementation of the TICT as well as for future research on the training's effectiveness are discussed.

Using 'Generic Abilities' to Foster Professionalism in a Post-Professional Program

Introduction To be successful in a profession, persons must have mastery of professional affective behaviors in addition to that profession’s core knowledge base. Health care delivery is more than just developing a good plan of care. The effective health care provider also will have good interpersonal and communication skills, have a commitment to learning, act professionally and responsibly, and have abilities in problem solving and critical thinking. But, these behaviors and characteristics are not taught directly in a professional program. This presentation will demonstrate how these ‘generic abilities’ have been incorporated in a post-professional program and how they can be used in other professional health care, professional development, and leadership programs. [See PDF for complete abstract]

The physician's perception of medical error and its application to the development of an educational training tool.

It is becoming clear that if we are to impact the rate of medical errors it will have to be done at the practicing physician level. The purpose of this project was to survey the attitude of physicians in Alabama concerning their perception of medical error, and to obtain their thoughts and desires for medical education in the area of medical errors. The information will be used in the development of a physician education program.

Hsp90 nuclear accumulation in quiescence is linked to chaperone function and spore development in yeast.

The 90-kDa heat-shock protein (Hsp90) operates in the context of a multichaperone complex to promote maturation of nuclear and cytoplasmic clients. We have discovered that Hsp90 and the cochaperone Sba1/p23 accumulate in the nucleus of quiescent Saccharomyces cerevisiae cells. Hsp90 nuclear accumulation was unaffected in sba1Delta cells, demonstrating that Hsp82 translocates independently of Sba1. Translocation of both chaperones was dependent on the alpha/beta importin SRP1/KAP95. Hsp90 nuclear retention was coincident with glucose exhaustion and seems to be a starvation-specific response, as heat shock or 10% ethanol stress failed to elicit translocation. We generated nuclear accumulation-defective HSP82 mutants to probe the nature of this targeting event and identified a mutant with a single amino acid substitution (I578F) sufficient to retain Hsp90 in the cytoplasm in quiescent cells. Diploid hsp82-I578F cells exhibited pronounced defects in spore wall construction and maturation, resulting in catastrophic sporulation. The mislocalization and sporulation phenotypes were shared by another previously identified HSP82 mutant allele. Pharmacological inhibition of Hsp90 with macbecin in sporulating diploid cells also blocked spore formation, underscoring the importance of this chaperone in this developmental program.

BMP-SIGNALING REGULATES A COMMON TRANSCRIPTIONAL PROGRAM TO CONTROL FACIAL FORM AND SKELETAL MORPHOGENESIS

Much of the craniofacial skeleton, such as the skull vault, mandible and midface, develops through direct, intramembranous ossification of the cranial neural crest (CNC) derived progenitor cells. Bmp-signaling plays critical roles in normal craniofacial development, and Bmp4 deficiency results in craniofacial abnormalities, such as cleft lip and palate. We performed an in depth analysis of Bmp4, a critical regulator of development, disease, and evolution, in the CNC. Conditional Bmp4 overexpression, using a tetracycline regulated Bmp4 gain of function allele, resulted in facial form changes that were most dramatic after an E10.5 Bmp4 induction. Expression profiling uncovered a signature of Bmp4 induced genes (BIG) composed predominantly of transcriptional regulators controlling self-renewal, osteoblast differentiation, and negative Bmp autoregulation. The complimentary experiment, CNC inactivation of Bmp2, Bmp4, and Bmp7, resulted in complete or partial loss of multiple CNC derived skeletal elements revealing a critical requirement for Bmp-signaling in membranous bone and cartilage development. Importantly, the BIG signature was reduced in Bmp loss of function mutants indicating similar Bmp-regulated target genes underlying facial form modulation and normal skeletal morphogenesis. Chromatin immunoprecipitation (ChIP) revealed a subset of the BIG signature, including Satb2, Smad6, Hand1, Gadd45g and Gata3 that was bound by Smad1/5 in the developing mandible revealing direct, Smad-mediated regulation. These data indicate that Bmp-signaling regulates craniofacial skeletal development and facial form by balancing self-renewal and differentiation pathways in CNC progenitors.

High-Resolution Ice Cores from US ITASE (West Antarctica): Development and Validation of Chronologies and Determination of Precision and Accuracy

Shallow ice cores were obtained from widely distributed sites across the West Antarctic ice sheet, as part of the United States portion of the International Trans-Antarctic Scientific Expedition (US ITASE) program. The US ITASE cores have been dated by annual-layer counting, primarily through the identification of summer peaks in non-sea-salt sulfate (nssSO(4)(2-)) concentration. Absolute dating accuracy of better than 2 years and relative dating accuracy better than 1 year is demonstrated by the identification of multiple volcanic marker horizons in each of the cores, Tambora, Indonesia (1815), being the most prominent. Independent validation is provided by the tracing of isochronal layers from site to site using high-frequency ice-penetrating radar observations, and by the timing of mid-winter warming events in stable-isotope ratios, which demonstrate significantly better than 1 year accuracy in the last 20 years. Dating precision to 1 month is demonstrated by the occurrence of summer nitrate peaks and stable-isotope ratios in phase with nssSO(4)(2-), and winter-time sea-salt peaks out of phase, with phase variation of < 1 month. Dating precision and accuracy are uniform with depth, for at least the last 100 years.

Individual Video Training iVT - Development of a National Infrastructure for Video-Supported Learning

Introduction: Video‐Supported Learning is particularly effective when it comes to skills and behaviors. Video registration of patient‐physician interviews, class room instruction or practical skills allow it to learners themselves, their peers, and their tutors to assess the quality of the learner's performance, to give specific feedback, and to make suggestions for improvement. Methods: In Switzerland, four pedagogical universities and two medical faculties joined to initiate the development of a national infrastructure for Video Supported Learning. The goal was to have a system that is simple to use, has most steps automated, provides the videos over the Internet, and has a sophisticated access control. Together with SWITCH, the national IT‐Support‐Organisation for Swiss Universities, the program iVT (Individual Video Training) was developed by integrating two preexisting technologies. The first technology is SWITCHcast, a podcast system. With SWITCHcast, videos are automatically uploaded to a server as soon as the registration is over. There the videos are processed and converted to different formats. The second technology is the national Single Logon System AAI (Authentification and Authorization Infrastructure) that enables iVT to link each video with the corresponding learner. The learner starts the registration with his Single Logon. Thus, the video can unambiguously be assigned. Via his institution's Learning Management System (LMS), the learner can access his video and give access to his video to peers and tutors. Results: iVT is now used at all involved institutions. The system works flawlessly. In Bern, we use iVT for the communications skills training in the forth and sixth year. Since students meet with patient actors alone, iVT is also used to certify attendance. Students are encouraged to watch the videos of the interview and the feedback of the patient actor. The offer to discuss a video with a tutor was not used by the students. Discussion: We plan to expand the use of iVT by making peer assessment compulsory. To support this, annotation capabilities are currently added to iVT. We also want to use iVT in training of practical skills, again for self as well as for peer assessment.  At present, we use iVT for quality control of patient actor's performance.