Remote control of CAR T cell therapies by thermal targeting

In 2017, the FDA approved two T cell therapies – Kymriah and Yescarta – for multiple relapsed or refractory hematological malignancies. In addition to providing new treatment options for patients with few viable alternatives, these approvals served as a watershed moment for an emerging class of cellular therapies based on T cells. As the use of these ‘living drugs’ grows, scientists, engineers, and clinicians continue to improve their therapeutic and safety profiles by developing new mechanisms to control immune cell activity in the body. Currently, the limited ability to control cytotoxicity within immunosuppressive tumors contributes to poor CAR T cell responses against solid malignancies. Systemic delivery of biologic drugs such as cytokines or checkpoint blockade inhibitors can augment T cell activity despite off-target toxicity in healthy tissues that narrows their therapeutic window. Thus, localizing the activity of adjuvant drugs and cytotoxicity of T cells to the tumor microenvironment constitutes an important goal for future immunotherapies. In this context, this thesis presents a remote control platform that enables localized control of engineered T cell activity via targeted thermal treatments. By re-engineering the cellular response to mild hyperthermia, I enhance the anti-tumor activity of therapeutic T cells through the heat-induced expression of immunostimulatory genes including Chimeric Antigen Receptors (CARs), cytokine superagonists, and Bispecific T cell Engagers (BiTEs). Aim 1 describes efforts to construct thermal gene switches using truncated sections of an endogenous heat shock promoter. It also introduces a photothermal method for targeted in vivo heating as well as pulsatile heating regimens which improve thermal tolerance and enhance switch activity in engineered cells compared to continuous heat treatments with an identical AUC. Aim 2 describes the design of synthetic thermal gene switches which exhibit lower basal activity and enhanced specificity for thermal cues compared to genomic sequences. In primary human T cells, these synthetic thermal gene switches enable control of critical effector functions such as proliferation and cytotoxicity against cancer cells. Using adoptive cell transfer in murine models of cancer, I demonstrate that photothermal control of engineered T cells improves their anti-tumor activity and treatment outcome. In the future, thermal control of engineered T cells could provide finer control of their in vivo activity and improve the safety and efficacy of next generation cellular therapies.Ph.D

Using Q-methodology to Determine Content of a Brochure on Lactation Consultants (in a Selected Area)

The Q-technique was utilized to determine content of a brochure on lactation consultants. The Q-sort which consisted of 40 cards was administered to 20 respondents--10 of who had breastfeeding experience and 10 of who did not have breastfeeding experience. The data collected and analyzed was used to determine the kind of information potential Stillwater, Oklahoma female clients of lactation consultants thought was important enough to be included in a brochure on lactation consultants.Mass Communciatio

The use of body language in public speaking

This research examined various methods of body language, gestures, and expressions used during public speeches. Numerous public speakers have used different techniques during their speeches to improve their presentations. Certain objectives of the research explain how body language can enhance public speech. The third objective was to examine how to prepare and implement various uses of body language into a public speech. Lastly, the research sought to identify the advantages of implementing different hand and arm movement methods while using assorted levels of voice ranges. The study was conducted by observing public speeches at expositions, seminars, government official public speeches, award acceptance speeches, and university professor lectures. Multiple prerecorded public speeches were also evaluated with the use of the internet. The primary results are that using different methods of hand gestures, arm movements, facial expressions, a strong voice, and maintaining a good posture significantly benefits public speeches. It was also established that preparing for a public speech by selecting the type of body language that will support your style of presentation will improve the success of your speech. The recommendation is that public speakers maintain a good posture and respect the audience while maintaining eye contact with all members attending the event. Additionally, the speakers should keep their hands and arm moving at all times while occasionally returning their hands to their waistline area and use various facial gestures and expressions to help entice the audience. Lastly, public speakers should use a strong voice during their speeches as it helps audience members focus during the presentation

Extraction, quantification and degree of polymerization of yacon (Smallanthus sonchifolia) fructans

Yacon tubers have been a distinguished alternative of fructans, drawing the attention of researchers and food industries. Since fructans are carbohydrate reserves storage can reduce their contents. Additionally, the type of extraction used can provide a higher yield of fructans. Therefore, it was necessary to study yacon storage and its influence on the extraction and quantification of fructans. Thus, the objective of this study was to evaluate three fructan extractions (water 75°C, water 95°C, ethanol 90°C) in yacon with 3 sizes (large, medium, small), stored for 20 days, at room temperature as well as to compare two quantification techniques. The three extractions can be used when fructans are quantified by high performance liquid chromatography (HPLC). For quantification by spectrometry, the best extraction method was ethanol at 90°C. Medium and small-sized tubers presented the highest contents of fructans that large tubers, and storage negatively influenced these contents. Fructan quantification by HPLC was higher than the spectrophotometric technique. All treatments showed a degree of polymerization in the range from 3 to 7, allowing numerous technological applications for fructans present in yacon.Key words: Fructooligosaccharides, storage, tuber size

Microfluidic Fabricated Liposomes for Nutlin-3a Ocular Delivery as Potential Candidate for Proliferative Vitreoretinal Diseases Treatment

Elisabetta Esposito,1 Elena Pozza,2 Catia Contado,1 Walter Pula,1 Olga Bortolini,3 Daniele Ragno,1 Sofia Toldo,3 Fabio Casciano,4 Agnese Bondi,1 Enrico Zauli,2 Paola Secchiero,4 Giorgio Zauli,3 Elisabetta Melloni4 1Department of Chemical, Pharmaceutical and Agricultural Sciences, University of Ferrara, Ferrara, I-44121, Italy; 2Department of Translational Medicine, University of Ferrara, Ferrara, I-44121, Italy; 3Department of Environmental Sciences and Prevention, University of Ferrara, Ferrara, I-44121, Italy; 4Department of Translational Medicine and LTTA Centre, University of Ferrara, Ferrara, I-44121, ItalyCorrespondence: Elisabetta Esposito; Elisabetta Melloni, Tel +39 0532 455230 ; +39 0532 455936, Email [email protected]; [email protected]: Proliferative vitreoretinal diseases (PVDs) represent a heterogeneous group of pathologies characterized by the presence of retinal proliferative membranes, in whose development retinal pigment epithelium (RPE) is deeply involved. As the only effective treatment for PVDs at present is surgery, we aimed to investigate the potential therapeutic activity of Nutlin-3a, a small non-genotoxic inhibitor of the MDM2/p53 interaction, on ARPE-19 cell line and on human RPE primary cells, as in vitro models of RPE and, more importantly, to formulate and evaluate Nutlin-3a loaded liposomes designed for ophthalmic administration.Methods: Liposomes were produced using an innovative approach by a microfluidic device under selection of different conditions. Liposome size distribution was evaluated by photon correlation spectroscopy and centrifugal field flow fractionation, while the liposome structure was studied by transmission electron microscopy and Fourier-transform infrared spectroscopy. The Nutlin-3a entrapment capacity was evaluated by ultrafiltration and HPLC. Nutlin-3a biological effectiveness as a solution or loaded in liposomes was evaluated by viability, proliferation, apoptosis and migration assays and by morphological analysis.Results: The microfluidic formulative study enabled the selection of liposomes composed of phosphatidylcholine (PC) 5.4 or 8.2 mg/mL and 10% ethanol, characterized by roundish vesicular structures with 150– 250 nm mean diameters. Particularly, liposomes based on the lower PC concentration were characterized by higher stability. Nutlin-3a was effectively encapsulated in liposomes and was able to induce a significant reduction of viability and migration in RPE cell models.Conclusion: Our results lay the basis for a possible use of liposomes for the ocular delivery of Nutlin-3a. Keywords: Nutlin-3a, liposomes, microfluidic, PVDs, CFF