Multiparameter flow cytometric analysis of C -reactive protein binding to human-derived cell lines and peripheral blood monocytes

C-reactive protein (CRP), a normally occurring human plasma protein may become elevated as much as 1,000 fold during disease states involving acute inflammation or tissue damage. Through its binding to phosphorylcholine in the presence of calcium, CRP has been shown to potentiate the activation of complement, stimulate phagocytosis and opsonize certain microorganisms. Utilizing a flow cytometric functional ligand binding assay I have demonstrated that a monocyte population in human peripheral blood and specific human-derived myelomonocytic cell lines reproducibly bind an evolutionarily conserved conformational pentraxin epitope on human CRP through a mechanism that does not involve its ligand, phosphorylcholine. ^ A variety of cell lines at different stages of differentiation were examined. The monocytic cell line, THP-1, bound the most CRP followed by U937 and KG-1a cells. The HL-60 cell line was induced towards either the granulocyte or monocyte pathway with DMSO or PMA, respectively. Untreated HL-60 cells or DMSO-treated cells did not bind CRP while cells treated with PMA showed increased binding of CRP, similar to U-937 cells. T cell and B-cell derived lines were negative. ^ Inhibition studies with Limulin and human SAP demonstrated that the binding site is a conserved pentraxin epitope. The calcium requirement necessary for binding to occur indicated that the cells recognize a conformational form of CRP. Phosphorylcholine did not inhibit the reaction therefore the possibility that CRP had bound to damaged membranes with exposed PC sites was discounted. ^ A study of 81 normal donors using flow cytometry demonstrated that a majority of peripheral blood monocytes (67.9 ± 1.3, mean ± sem) bound CRP. The percentage of binding was normally distributed and not affected by gender, age or ethnicity. Whole blood obtained from donors representing a variety of disease states showed a significant reduction in the level of CRP bound by monocytes in those donors classified with infection, inflammation or cancer. This reduction in monocyte populations binding CRP did not correlate with the concentration of plasma CRP. ^ The ability of monocytes to specifically bind CRP combined with the binding reactivity of the protein itself to a variety of phosphorylcholine containing substances may represent an important bridge between innate and adaptive immunity. ^

Immunophylogenetic aspects of a gorgonian coral

One goal of comparative immunology is to derive inferences about evolutionary pathways in the development of immune-defense systems. Almost 700 million years ago, a major divergence occurred in the phylogeny of animals, spitting all descendants into either the protostome or deuterostome (includes vertebrates) lineages. Genes have evolved independently along these lineages for that amount of time. Cnidarians originated before that divergence event, and can hold clues as to which immune response genes are homologous to both lineages. This work uses the gorgonian coral, Swiftia exserta, for two major reasons: (1) because of their phylogenetic position, corals are an important animal model in studies concerning the phylogeny of immune-response genes, and (2) nothing is known about the genes controlling immunocompetence in corals. The work described here has important implications in both innate and adaptive immunity. ^ The vertebrate complement system is a major component of innate immunity. C3 is a critical component of the three pathways of complement. Because of its opsonic properties, a C3-like protein is expected to have evolved early. However, currently available data suggests that complement-like components are unique to the deuterostome lineage. This work describes the cloning and characterization of a C3-like gene from S. exserta. The deduced polypeptide sequence reveals conservation of multiple, functionally critical, sites while sharing physiochemical and structural properties with the complement components C3/C4/C5. ^ Antigen processing, via intracellular enzymatic proteasomes, is a major requirement of vertebrate adaptive immunity. These organelles have a catalytic core, through which pass intracellular proteins for degradation into peptides presentable to the immune system. LMP 7 is one component of the paralogous “immuno-proteasome”. LMP 7 is a paralog of the ubiquitous LMP X, but is restricted to vertebrates. While LMP 7 is absent in the coral, this work describes a coral LMP X gene. Phylogenetic analyses, along with hydropathy profiling of a critical portion of the invertebrate and vertebrate paralogous genes, suggests that some invertebrates have two diverging LMP X genes. In some cases, one LMP X protein shares characteristics with vertebrate LMP 7. This work presents new evidence for how the LMP X and 7 genes evolved. ^

Cooperative and adaptive medium access control in multi-hop wireless networks

Due to low cost and easy deployment, multi-hop wireless networks become a very attractive communication paradigm. However, IEEE 802.11 medium access control (MAC) protocol widely used in wireless LANs was not designed for multi-hop wireless networks. Although it can support some kinds of ad hoc network architecture, it does not function efficiently in those wireless networks with multi-hop connectivity. Therefore, our research is focused on studying the medium access control in multi-hop wireless networks. The objective is to design practical MAC layer protocols for supporting multihop wireless networks. Particularly, we try to prolong the network lifetime without degrading performances with small battery-powered devices and improve the system throughput with poor quality channels. ^ In this dissertation, we design two MAC protocols. The first one is aimed at minimizing energy-consumption without deteriorating communication activities, which provides energy efficiency, latency guarantee, adaptability and scalability in one type of multi-hop wireless networks (i.e. wireless sensor network). Methodologically, inspired by the phase transition phenomena in distributed networks, we define the wake-up probability, which maintained by each node. By using this probability, we can control the number of wireless connectivity within a local area. More specifically, we can adaptively adjust the wake-up probability based on the local network conditions to reduce energy consumption without increasing transmission latency. The second one is a cooperative MAC layer protocol for multi-hop wireless networks, which leverages multi-rate capability by cooperative transmission among multiple neighboring nodes. Moreover, for bidirectional traffic, the network throughput can be further increased by using the network coding technique. It is a very helpful complement for current rate-adaptive MAC protocols under the poor channel conditions of direct link. Finally, we give an analytical model to analyze impacts of cooperative node on the system throughput. ^

High natural gene expression variation in the reef-building coral Acropora millepora: potential for acclimative and adaptive plasticity

Background: Ecosystems worldwide are suffering the consequences of anthropogenic impact. The diverse ecosystem of coral reefs, for example, are globally threatened by increases in sea surface temperatures due to global warming. Studies to date have focused on determining genetic diversity, the sequence variability of genes in a species, as a proxy to estimate and predict the potential adaptive response of coral populations to environmental changes linked to climate changes. However, the examination of natural gene expression variation has received less attention. This variation has been implicated as an important factor in evolutionary processes, upon which natural selection can act. Results: We acclimatized coral nubbins from six colonies of the reef-building coral Acropora millepora to a common garden in Heron Island (Great Barrier Reef, GBR) for a period of four weeks to remove any site-specific environmental effects on the physiology of the coral nubbins. By using a cDNA microarray platform, we detected a high level of gene expression variation, with 17% (488) of the unigenes differentially expressed across coral nubbins of the six colonies (jsFDR-corrected, p < 0.01). Among the main categories of biological processes found differentially expressed were transport, translation, response to stimulus, oxidation-reduction processes, and apoptosis. We found that the transcriptional profiles did not correspond to the genotype of the colony characterized using either an intron of the carbonic anhydrase gene or microsatellite loci markers. Conclusion: Our results provide evidence of the high inter-colony variation in A. millepora at the transcriptomic level grown under a common garden and without a correspondence with genotypic identity. This finding brings to our attention the importance of taking into account natural variation between reef corals when assessing experimental gene expression differences. The high transcriptional variation detected in this study is interpreted and discussed within the context of adaptive potential and phenotypic plasticity of reef corals. Whether this variation will allow coral reefs to survive to current challenges remains unknown.

A multimodal and adaptive real-time human -computer interface for peoples with severe motor disability

This dissertation introduces the design of a multimodal, adaptive real-time assistive system as an alternate human computer interface that can be used by individuals with severe motor disabilities. The proposed design is based on the integration of a remote eye-gaze tracking system, voice recognition software, and a virtual keyboard. The methodology relies on a user profile that customizes eye gaze tracking using neural networks. The user profiling feature facilitates the notion of universal access to computing resources for a wide range of applications such as web browsing, email, word processing and editing. ^ The study is significant in terms of the integration of key algorithms to yield an adaptable and multimodal interface. The contributions of this dissertation stem from the following accomplishments: (a) establishment of the data transport mechanism between the eye-gaze system and the host computer yielding to a significantly low failure rate of 0.9%; (b) accurate translation of eye data into cursor movement through congregate steps which conclude with calibrated cursor coordinates using an improved conversion function; resulting in an average reduction of 70% of the disparity between the point of gaze and the actual position of the mouse cursor, compared with initial findings; (c) use of both a moving average and a trained neural network in order to minimize the jitter of the mouse cursor, which yield an average jittering reduction of 35%; (d) introduction of a new mathematical methodology to measure the degree of jittering of the mouse trajectory; (e) embedding an onscreen keyboard to facilitate text entry, and a graphical interface that is used to generate user profiles for system adaptability. ^ The adaptability nature of the interface is achieved through the establishment of user profiles, which may contain the jittering and voice characteristics of a particular user as well as a customized list of the most commonly used words ordered according to the user's preferences: in alphabetical or statistical order. This allows the system to successfully provide the capability of interacting with a computer. Every time any of the sub-system is retrained, the accuracy of the interface response improves even more. ^

Immunomodulation by shark cartilage extracts

The immune system is composed of innate and adaptive mechanisms. Innate immune responses are significantly modulated by immunomodulatory factors that act through the induction of specific patterns of cytokine production in responding cells. Human leukocytes have been shown to respond to substance(s) present in acid extracts of commercial shark cartilage (SC). Shark cartilage is a food supplement taken by consumers as a prophylaxis and for the treatment of conditions ranging from arthritis to cancer. No reliable scientific evidence in the literature supports the alleged usefulness of shark cartilage supplements, but their use remains popular. Cartilage extracts exhibit immunomodulatory properties by inducing various inflammatory, Th1-type cytokines and potent chemokines in human peripheral blood leukocytes (HPBL) in vitro. The objectives of the study were to (1) to determine the nature of the active component(s), (2) to define the scope of cellular response to SC extract, and (3) to elucidate the molecular mechanisms underlying bioactivity. Results showed that there are at least two cytokine-inducing components which are acid stable. One anionic component has been identified as a small (14-21 kDa) glycoprotein with at least 40% carbohydrate content. Shark cartilage stimulated HPBL to produce cytokines resembling an inflammatory, Th1 polarized response. Leukocyte-specific responses consist of both initial cytokine responses to SC directly (i.e., TNF-α) and secondary responses such as the IFN-γ response by lymphocytes following initial SC stimulation. Response of RAW cells, a murine macrophage cell line, indicated that TNF-á could be induced in macrophages of another mammalian species in the absence of other cell types. The results suggest that the human monocyte/macrophage is most likely to be the initial responding cell to SC stimulation. Stimulation of cells appears to engage at least one ligand-receptor interaction with TLR 4, although the role of TLR 2 cannot be ruled out. Initial activation is likely followed by the activation of the JNK and p38 MAPK signal transduction pathways resulting in activation, release, and translocation of transcription factor nuclear factor κB (Nf-κB). This dissertation research study represents the first in-depth study into characterizing the bioactive component(s) of commercial shark cartilage responsible for its immunomodulating properties and defining cellular responses at the molecular level.

The development, application, and implications of a strategy for reflective learning from experience

The problem on which this study focused was individuals' reduced capacity to respond to change and to engage in innovative learning when their reflective learning skills are limited. In this study, the preceding problem was addressed by two primary questions: To what degree can mastery of a strategy for reflective learning be facilitated as a part of an academic curriculum for professional practitioners? What impact will mastery of this strategy have on the learning style and adaptive flexibility of adult learners? The focus of the study was a direct application of human resource development technology in the professional preparation of teachers. The background of the problem in light of changing global paradigms and educational action orientations was outlined and a review of the literature was provided. Roots of thought for two key concepts (i.e., learning to learn from experience and meaningful reflection in learning) were traced. Reflective perspectives from the work of eight researchers were compared. A meta-model of learning from experience drawn from the literature served as a conceptual framework for the study. A strategy for reflective learning developed from this meta-model was taught to 109 teachers-in-training at Florida International University in Miami, Florida. Kolb's Adaptive Style Inventory and Learning Style Inventory were administered to the treatment group and to two control groups taught by the same professor. Three research questions and fourteen hypotheses guided data analysis. Qualitative review of 1565 personal documents generated by the treatment group indicated that 77 students demonstrated "double-loop" learning, going beyond previously established limits to perception, understanding, or action. The mean score for depth of reflection indicated "single-loop" learning with "reflection-in-action" present. The change in the mean score for depth of reflection from the beginning to end of the study was statistically significant (p $<$.05). On quantitative measures of adaptive flexibility and learning style, with two exceptions, there were no significant differences noted between treatment and control groups on pre-test to post-test differences and on post-test mean scores adjusted for pre-test responses and demographic variables. Conclusions were drawn regarding treatment, instrumentation, and application of the strategy and the meta-model. Implications of the strategy and the meta-model for research, for education, for human resource development, for professional practice, and for personal growth were suggested. Qualitative training materials and Kolb's instruments were provided in the appendices.

Optimization of adaptive traffic signal control with transit signal priority at isolated intersections using parallel genetic algorithms

Optimization of adaptive traffic signal timing is one of the most complex problems in traffic control systems. This dissertation presents a new method that applies the parallel genetic algorithm (PGA) to optimize adaptive traffic signal control in the presence of transit signal priority (TSP). The method can optimize the phase plan, cycle length, and green splits at isolated intersections with consideration for the performance of both the transit and the general vehicles. Unlike the simple genetic algorithm (GA), PGA can provide better and faster solutions needed for real-time optimization of adaptive traffic signal control. ^ An important component in the proposed method involves the development of a microscopic delay estimation model that was designed specifically to optimize adaptive traffic signal with TSP. Macroscopic delay models such as the Highway Capacity Manual (HCM) delay model are unable to accurately consider the effect of phase combination and phase sequence in delay calculations. In addition, because the number of phases and the phase sequence of adaptive traffic signal may vary from cycle to cycle, the phase splits cannot be optimized when the phase sequence is also a decision variable. A "flex-phase" concept was introduced in the proposed microscopic delay estimation model to overcome these limitations. ^ The performance of PGA was first evaluated against the simple GA. The results show that PGA achieved both faster convergence and lower delay for both under- or over-saturated traffic conditions. A VISSIM simulation testbed was then developed to evaluate the performance of the proposed PGA-based adaptive traffic signal control with TSP. The simulation results show that the PGA-based optimizer for adaptive TSP outperformed the fully actuated NEMA control in all test cases. The results also show that the PGA-based optimizer was able to produce TSP timing plans that benefit the transit vehicles while minimizing the impact of TSP on the general vehicles. The VISSIM testbed developed in this research provides a powerful tool to design and evaluate different TSP strategies under both actuated and adaptive signal control. ^

Best Practices: Lessons Learned by a South Florida Non-Profit Community Based Organization while Designing and Implementing a Career Exploration Evidence Informed Framework in Urban Communities

All A’s was designed to support of the agency’s family strengthening initiatives in South Florida. All A’s uses evidence informed strategies poised to be an inclusive curriculum that teaches self-determination and adaptive behavior skills. The framework incorporates problem based learning and adult learning theory and follows the Universal Design for Learning. Since 2012, the agency has served over 8500 youth and 4,000 adults using the framework. The framework addresses educational underachievement and career readiness in at risk populations. It is used to enhance participants AWARENESS of setting SMART goals to achieve future goals and career aspirations. Participants are provided with ACCESS to resources and opportunities for creating and implementing an ACTION plan as they pursue and ACHIEVE their goals. All A’s promotes protective factors and expose youth to career pathways in Science, Technology, Engineering and Math (STEM) related fields. Youth participate in college tours, job site visits, job shadowing, high school visits, online college and career preparation assistance, service learning projects, STEM projects, and the Winning Futures© mentoring program. Adults are assisted with résumé development; learn job search strategies, interview techniques, job shadowing experiences, computer and financial literacy programs. Adults and youth are also given the opportunity to complete industry-recognized certifications in high demand industries (food service, general labor, and construction), and test preparation for the General Educational Development Test.

World-Class Service

In the discussion - World-Class Service - by W. Gerald Glover, Associate Professor, Restaurant, Hotel and Resort Management at Appalachian State University and Germaine W. Shames, Hilton International, New York, Glover and Shames initially state: “Providing world-class service to today's traveler may be the key for hospitality managers in the current competitive market. Although an ideal, this type of service provides a mandate for culturally aware managers. The authors provide insight into several areas of cultures in collision.” Up to the time this essay is written, the authors point to a less-than-ideal level of service as being the standard in the hospitality industry and experience. “Let's face it - if we're ever to resurrect service, it will not be by going back to anything,” Glover and Shames exclaim. “Whatever it was we did back then has contributed to the dilemma in which we find ourselves today, handicapped by a reactive service culture in an age that calls for adaptiveness and global strategies,” the authors fortify that thought. In amplifying the concept of world-class service Glover and Shames elaborate: “World-class service is an ideal. Proactive and adaptive, world-class service feels equally right to the North American dignitary occupying the Presidential Suite, and the Japanese tourist staying in a standard room in the same hotel.” To bracket that model the authors offer: “At a minimum, it is service perceived by each customer as appropriate and adequate. At its best, it may also make the customer feel at home, among friends, or pampered. Finally, it is service as if culture matters,” Glover and Shames expand and capture the rule of world-class service. Glover and Shames consider the link between cultures and service an imperative one. They say it is a principle lost on most hospitality managers. “Most [managers] have received service management education in the people are people school that teaches us to disregard cultural differences and assume that everyone we manage or serve is pretty much like ourselves,” say Glover and Shames. “Is it any wonder that we persist in setting service standards, marketing services, and managing service staff not only as if culture didn't matter, but as if it didn't exist?!” To offer legitimacy to their effort Glover and Shames present the case of the Sun and Sea Hotel, a 500-room first class hotel located on the outskirts of the capital city of a small Caribbean island nation. It is a bit difficult to tell whether this is a dramatization or a reality. It does, however, serve to illustrate their point in regard to management’s cognizance, or lack thereof, of culture when it comes to cordial service and guest satisfaction. Even more apropos is the tale of the Palace Hotel, “…one of the grande dames of hospitality constructed in the boom years of the 1920s in a mid-sized Midwestern city in the United States.” The authors relate what transpired during its takeover in mid-1980 by a U.S.-based international hotel corporation. The story makes for an interesting and informative case study.

Identity as a moderator of guided -self change treatment outcomes with adolescents

Despite recent calls to examine the possible moderating effects of developmental variables on treatment outcomes with adolescents, most research has continued to focus on age as a main developmental variable. Building upon the theoretical notion that identity development is a central task of adolescence, this study investigated whether adolescents' response to an alcohol and other drug (AOD) use reduction intervention, Guided-Self Change (GSC), was influenced by their approach to self-defining questions and situations. While past research has established associations between maladaptive identity development and alcohol and other drug use, very little research attention has been given to the potential relationship that may exist between identity variables and AOD treatment response. Given GCS's promotion of self-reflective, self-exploring, and problem-focused coping skills as a way of addressing AOD problems, it was hypothesized that adolescents with positive identity development (i.e., greater identity coherence and adaptive identity processing styles) would respond more positively to GCS's change producing therapeutic techniques than their counterparts. This hypothesis was tested with an ethnically diverse sample of 134 adolescents between the ages of 14-18 years who were randomly assigned to either a GSC intervention or a comparison control group. Results revealed that identity development was significantly associated with AOD use, such that adolescents with diffused/avoidant styles and high levels of distress over identity-related issues used more alcohol at the baseline assessment than those with more positive identity development. Results also indicated that while identity distress or identity coherence did not moderate GSC treatment outcomes, high scores for a diffused/avoidant style did moderate treatment outcomes and thus proved to be a significant predictor of treatment response. These results indicate that identity development influences adolescents' alcohol use and that while use of certain approaches to self-defining questions does not predict treatment outcomes, high use for negative approaches does moderate intervention-related change. ^

Supporting Students with Intellectual Disabilities in Higher Education through Mentoring

Federal legislation is now making it possible for students with Intellectual Disabilities (ID) to enroll in university programs through inclusive transition models. The need for more knowledge in the workplace, in addition to newly acquired opportunities, draws students with ID to college campuses. With this increase of student in postsecondary education settings, it is necessary to examine the types of supports needed to ensure their success. Peer mentorship has been identified as a crucial aspect of support for academic and social development. This paper identifies the skills mentors use during their sessions and outlines challenges with mentoring partnerships in postsecondary transition programs. Data analysis revealed major themes related to inclusion, self-determination, and adaptive behavior skills.

Evaluating the Effectiveness of Discrete Trial Procedures for Teaching Receptive Discrimination to Children with Autism Spectrum Disorders

Research has found that children with autism spectrum disorders (ASD) show significant deficits in receptive language skills (Wiesmer, Lord, & Esler, 2010). One of the primary goals of applied behavior analytic intervention is to improve the communication skills of children with autism by teaching receptive discriminations. Both receptive discriminations and receptive language entail matching spoken words with corresponding objects, symbols (e.g., pictures or words), actions, people, and so on (Green, 2001). In order to develop receptive language skills, children with autism often undergo discrimination training within the context of discrete trial training. This training entails teaching the learner how to respond differentially to different stimuli (Green, 2001). It is through discrimination training that individuals with autism learn and develop language (Lovaas, 2003). The present study compares three procedures for teaching receptive discriminations: (1) simple/conditional (Procedure A), (2) conditional only (Procedure B), and (3) conditional discrimination of two target cards (Procedure C). Six children, ranging in age from 2-years-old to 5-years-old, with an autism diagnosis were taught how to receptively discriminate nine sets of stimuli. Results suggest that the extra training steps included in the simple/conditional and conditional only procedures may not be necessary to teach children with autism how to receptively discriminate. For all participants, Procedure C appeared to be the most efficient and effective procedure for teaching young children with autism receptive discriminations. Response maintenance and generalization probes conducted one-month following the end of training indicate that even though Procedure C resulted in less training sessions overall, no one procedure resulted in better maintenance and generalization than the others. In other words, more training sessions, as evident with the simple/conditional and conditional only procedures, did not facilitate participants’ ability to accurately respond or generalize one-month following training. The present study contributes to the literature on what is the most efficient and effective way to teach receptive discrimination during discrete trial training to children with ASD. These findings are critical as research shows that receptive language skills are predictive of better outcomes and adaptive behaviors in the future.

Evaluating the effectiveness of discrete trial procedures for teaching receptive discrimination to children with autism spectrum disorders

Research has found that children with autism spectrum disorders (ASD) show significant deficits in receptive language skills (Wiesmer, Lord, & Esler, 2010). One of the primary goals of applied behavior analytic intervention is to improve the communication skills of children with autism by teaching receptive discriminations. Both receptive discriminations and receptive language entail matching spoken words with corresponding objects, symbols (e.g., pictures or words), actions, people, and so on (Green, 2001). In order to develop receptive language skills, children with autism often undergo discrimination training within the context of discrete trial training. This training entails teaching the learner how to respond differentially to different stimuli (Green, 2001). It is through discrimination training that individuals with autism learn and develop language (Lovaas, 2003). The present study compares three procedures for teaching receptive discriminations: (1) simple/conditional (Procedure A), (2) conditional only (Procedure B), and (3) conditional discrimination of two target cards (Procedure C). Six children, ranging in age from 2-years-old to 5-years-old, with an autism diagnosis were taught how to receptively discriminate nine sets of stimuli. Results suggest that the extra training steps included in the simple/conditional and conditional only procedures may not be necessary to teach children with autism how to receptively discriminate. For all participants, Procedure C appeared to be the most efficient and effective procedure for teaching young children with autism receptive discriminations. Response maintenance and generalization probes conducted one-month following the end of training indicate that even though Procedure C resulted in less training sessions overall, no one procedure resulted in better maintenance and generalization than the others. In other words, more training sessions, as evident with the simple/conditional and conditional only procedures, did not facilitate participants’ ability to accurately respond or generalize one-month following training. The present study contributes to the literature on what is the most efficient and effective way to teach receptive discrimination during discrete trial training to children with ASD. These findings are critical as research shows that receptive language skills are predictive of better outcomes and adaptive behaviors in the future. ^