MECHANISTIC AND FUNCTIONAL CHARACTERIZATION OF B AND T LYMPHOCYTE ATTENUATOR (BTLA) IN CD8 TUMOR INFILTRATING LYMPHOCYTES

This dissertation project focused on understanding the functional role of BTLA on CD8+ Tumor Infiltrating Lymphocytes (TIL) from metastatic melanoma patients. Clinical trials of adoptive T-cell therapy (ACT) using autologous ex vivo expanded TIL have demonstrated the great potential of this immunotherapy with an overall clinical response rate 40-50% for stage IV metastatic melanoma patients. We have investigated a number of biomarkers in both the infused TIL and the tumor microenvironment for their association with clinical response. Surprisingly, a subset of CD8+ TIL expressing the co-inhibitory molecule BTLA (B-and-T lymphocyte attenuator) was highly associated with clinical response, while expression of other co-inhibitory molecules such as PD-1, TIM-3, and Lag3 did not associate with response. BTLA is expressed by T cells, B cells, and NK cells and serves as a T cell differentiation maker whereby high expression of BTLA associates with less differentiated T cell phenotype. While the suppressive function of the ITIM and ITSM motifs of BTLA are well described, the Grb2 motif’s function remains understudied. In this study, we sought to determine the functional characteristics of the CD8+BTLA+TIL subset and define the contribution of the Grb2 motif of BTLA in T cell co-stimulation. We have uncovered a survival advantages of the BTLA+ subset that allows for serial killing of target tumor cells, which may explain our previous correlation between this subset and response to TIL ACT. BTLA-HVEM interaction during T cell activation led to the specific activation of SRC kinase. In addition, our results unveiled a role for the BTLA-associated Grb2-binding motif in T cell proliferation and IL-2 production following TCR engagement that was independent of the inhibitory function of ITIM/ITSM motifs. Overall, our study first unveil the dual role of BTLA as both a co-stimulatory and co-inhibitory molecule. The integration of the positive and negative signals transduced by BTLA promotes IL-2 secretion while reducing certain effector function of T cell. Altogether, the combination of both BTLA signaling and inherent attributes of less differentiated T cells could promote T cell survival, persistence, and anti-tumor function

Development and evaluation of glass fiber-reinforced composite/wood railroad crossties

Railroad crossties are strengthened by wrapping glass fiber reinforced composites (GFRC). In addition, durability of crossties has been found to have enhanced by reinforcing them with GFRC wraps. In order to reduce the cost, GFRC wraps are not used to wrap the entire length of a crosstie, but only in the region that holds steel plates. The objective of this research is to select compatible adhesive between creosote treated wood and glass fabric, and study the performance of wrapped wooden crossties.;Five primer/resin combinations (Resorcinol Formaldehyde group) were screened for strength and durability of treated wood crossties. To evaluate stiffness and durability of wrapped samples under natural environmental conditions (simulated by six cycles aging), half scale wooden crossties (3 in. x 4 in. x 42 in.) were wrapped with glass fabric at midspan over a length of 20 in. before subjecting them to three point bending test. These tests were conducted on half scale samples, before and after accelerated aging conditions.;To study the performance of GFRC full scale crossties, fatigue testing was carried out on full scale non-wrapped and GFRC wrapped specimens. Full scale crossties were wrapped over the rail-seat zones and then embedded in ballast under flexural fatigue. The experimental deflections and bending moment correlated well with the analytical values using the bending theory of beams on elastic foundation. After evaluating the performance of GFRC wrapped crossties in the laboratory, the GFRC full scale crossties were manufactured and installed in the field. Preliminary field results of full scale GFRC wrapped crossties are reported herein

Theoretical and experimental analysis of FRP bridge decks under thermal loads

The temperature difference between the top and bottom of a fiber reinforced polymer (FRP) composite deck (120°F) is nearly three times that of conventional concrete decks (40°F). The large temperature difference is attributed to low thermal conductivity of FRP material and low thermal mass due to hollow core. Thermal response studies have been conducted for FRP bridge decks under thermal fluctuations and temperature difference across the deck depth.;In this study, thermal tests were conducted on two FRP bridge deck modules (4&inches; and 8&inches; deep decks) in the laboratory by heating or cooling at the top surface of FRP deck (i.e., room temperature at bottom surface). The FRP deck boundaries were either four free boundaries (FFFF) or two opposites boundaries being free while the remaining two were simply supported (SSFF). Deflections and strains were recorded at different location under thermal loads. Closed form solutions with first term approximation were derived using the plate bending theory using Macro Approach and Navier-Levy method for SSFF boundary conditions. Theoretical results (using Macro Approach, Navier-Levy, and FEM) were compared with experimental results. In addition, thermal responses of two FRP deck bridges (i.e., Market Street Bridge and Wickwire Run Bridge) under thermal difference between the deck top and bottom were evaluated after establishing coefficients of thermal expansion (CTE) of both FRP decks. The laboratory test data indicated that the FRP deck exhibits a hogging effect (upward convexity) when it was subjected to positive temperature difference (i.e., Ttop \u3e Tbottom, heating test) and a sagging effect when it was subjected to negative temperature difference (i.e., Ttop \u3c Tbottom , cooling test). Deflections increased with increasing magnitude of temperature difference. The positive strain (expansion) and compressive stress were induced in the FRP deck when temperature of FRP decks was increased by direct exposure to Sun light. Partial deck restraint, provided by steel stringer, resulted in partially induced stresses. The transient thermal stresses could be as high as 45% of the allowable stress of FRP decks and the transient thermal strain could be as high as 86% of allowable strain of the FRP bridge deck modules

FUNCTIONAL CHARACTERIZATION OF THE VIRAL FLICE INHIBITORY PROTEIN OF RHESUS MONKEY RHADINOVIRUS

Rhesus monkey rhadinovirus (RRV) is a gamma-2 herpesvirus closely related to Kaposi's sarcoma-associated herpesvirus (KSHV). KSHV is associated with several malignant diseases, including Kaposi's sarcoma, primary effusion lymphoma and multicentric Castleman's disease. Here we found that RRV viral FLICE inhibitory protein (vFLIP) inhibits apoptosis. In HeLa cells with vFLIP expression, cleavage of poly [ADP-ribose] polymerase 1 (PARP-1) and activities of caspase 3, 7, and 9 were much lower than controls. RRV vFLIP was able to enhance cell survival under starved condition or apoptosis induction. After apoptosis induction, autophagosome formation was enhanced in cells with vFLIP expression and when autophagy was inhibited, these cells underwent apoptosis. Moreover, RRV latent infection of BJAB B-lymphoblastoid cells protects the cells against apoptosis. Knockdown of vFLIP expression in the RRV-infected BJAB cells with siRNA abolished the protection against apoptosis. These findings indicate that RRV vFLIP protects cells against apoptosis by enhancing autophagosome formation

Characterization of natural organic matter and precursors to trihalomethanes using spectral flourescence signatures

Disinfection is an essential process to kill pathogens (i.e., disease causing organisms) in source water during the production of drinking water. Chlorine is most widely used disinfectant because it is effective, affordable, and also provides chlorine residual to ensure that the water is safe through the distribution system. Nonetheless, chlorine reacts with Natural Organic Matter (NOM) and forms potentially carcinogenic Disinfection By-products (DBPs). The major chlorination DBPs are dominantly Trihalomethanes (THMs). However, not all organic compounds are equally reactive to THMs formation. NOM in water samples collected from the Delaware & Raritan Canal and its tributaries (Central New Jersey) was isolated by resin adsorption into six fractions: Hydrophobic acid (HPOA), Hydrophobic neutral (HPON), Hydrophobic base (HPOB), Hydrophilic acid (HPIA), Hydrophilic neutral (HPIN), and Hydrophilic base (HPIB). HPIN, HPON, and HPOA were the major fractions in most of samples. Moreover, the fractions\u27 seven-day THMs Formation Potentials (THMFP) were determined HPOA was found to be the most reactive fraction to THMs formation in addition to being one of the most abundant fractions in the source water. Additionally, the six fractions were also characterized by fluorescence spectroscopy to obtain three-dimensional fluorescence spectra. The spectra shape and peak locations are unique characteristics of organic compounds and also called Spectral Fluorescence Signature (SFS). The SFS is the total sum of emission intensity of a sample at different excitation wavelengths, recorded as a matrix of fluorescent intensity in coordinates of excitation and emission wavelengths. Among the six fractions, HPOA spectra were large and the peak intensity was also high. Therefore, fluorescence spectroscopy could be a promising technique for characterization of HPOA fraction or THMs precursors in the source water.Although a large number of intensities are related to THMs precursors, many of them are highly correlated by nature. Principle component analysis was then used to transform the fluorescence intensities into independent parameters called Principle Components (PCs). Best Subset Algorithm was performed to select the most important PCs for the prediction of THMFP by multiple linear regression. The prediction of THMFP using SFS is a rapid, inexpensive, reagent-free technique and thus can be used for optimization of water treatment processes

Trifluoroiodomethane as an environmentally friendly gas for water patterning by plasma etching process

Trifluoroiodomethane (CF3I), a non-global warming gas, has been investigated with study as a substitute for typical CFC etchants, such as CF4 and C2F6, used in wafer pattering technology. This investigation was carried out by exposing dielectric films of silicon oxide (SiO2) and silicon nitride (Si3N4) in CF3I and C2F6/O2 (used as a reference) plasma environments. The etch rate of these films was ascertained as function of applied rf power, etchant gas flow rate, reaction chamber operating pressure, and O2 to CF3I ratio. Upon increasing power, the etch rates of SiO2 and Si3N4 by both CF3I and C2F6/O2 were increased. The study of flow rate showed various results depending on types of etchants and dielectric film materials. According to the processing conditions, there were two limits in etching with CF3I. First, at the pressure higher than 50 mTorr, etching of\u27 Si3N4 can produce polymer film on sample surface. Secondly, at O2/CF3I ratio higher than 0.17, etching of both dielectric films can result in generating reddish particles all over the sample and the reaction chamber side wall

Art as knowledge: situational geography of the borderlines

Das Buch ist eine Sammlung von Reisenotizen, Tagebüchern, Meinungsäußerungen von Aktivisten, Eventberichten und enzyklopädischen Zusammenfassungen über die Grenzverwaltung beteiligten Organisationen. Es ist eine Art faszinierender Zwischenablage mit Aufsätzen in einem 'lockeren' Format und einer maximalen Länge von fünf Seiten. Dieses Buch verweist auf die Vielzahl von Ebenen, von denen aus die Themen der Grenzen und Grenzübergange angegangen werden könnten. Man könnte dieses Buch dem Gebiet der Situationsgeografie zuordnen, welche den Zusammenhang der Interaktion von menschlichen und natürlichen Einflüssen mit Blick auf verschiedene Standorte untersucht; in diesem Fall – die Grenzen. Diese einzigartige Mischung bietet den Lesern die Möglichkeit die grenzbezogenen Themen zusammen zu denken, die ansonsten vielleicht nicht in einen Dialog miteinander geraten wären. Eine spezifische Analyse dieses Netzwerks von Ideen ist allerdings nicht vorgegeben, so dass der Leser aufgefordert wird, das Verbindende selbst zu entdecken. Ohne ein tiefgreifendes theoretisches Fundament, basiert dieses Buch vor allem auf persönlichen Erfahrungen, Kunstprojekten und pointiert politischen Statements der Autoren. Es ist eine aufrichtige Einladung das Potential des Themas zu erkunden, gleichwohl angeregt durch die Kritik an den bestehenden Systemen bzw. dem politischen Zustand der EU-Grenzverwaltung.The book is a collection of travel notes, diaries, expressions of activists’ opinions, event reports, and encyclopedia excerpts on the organizations involved in border management. It is a type of intriguing clipboard, one in which articles are given more relaxed formats with small-cap titles and a maximum length of five pages. This book creates the potential for a multiplicity of layers through which the topic of borders and border crossing could be approached. One could associate this book with the field of situational geography, the study of interaction of human and natural complexities organized around the descriptive research of different sites, in this case – borders. This unique mix offers readers an opportunity to connect border-related themes that perhaps would not otherwise have been integrated into a dialog with each other. An analysis of this network of ideas is not predetermined, however, so the reader is encouraged to discover the potential themes him or herself. Without a profound theoretical framework, this book is mainly based on personal experiences, art projects, and the well-stated political opinions of the contributors. It is a genuine invitation to explore the potential of the topic, criticizing the existing systems and questioning the state of politics that prevails in border management, particularly in the EU