Characterization of Fc receptor family proteins in vaginal and endocervical epithelia

In the age of highly active antiretroviral therapy (HAART), patients infected with Human Immunodeficiency Virus Type 1 (HIV-1) are now living significantly healthier and longer lives. However, HIV prevention and cure still remain significant challenges. Globally, women face specific barriers to using and accessing both female and male condoms, the primary method recommended by the World Health Organization (WHO) to prevent sexual transmission of HIV-1. Although HAART treatment as prevention (TasP) of HIV has shown promising preliminary results, poor economic feasibility of the method in resource poor settings has yet to be resolved. Since women carry over 50% of the disease burden, there is a significant need for the development of a female-controlled method of prevention. One such approach is the reformulation of topical vaginal microbicides. Our laboratory is developing HIV-targeted microbicide formulations that utilize highly specific, broadly neutralizing anti-HIV antibodies (bNAbs). The purpose of my research project was to characterize Fc receptor expression in epithelial cell models of the lower female genital tract with a particular focus on the neonatal Fc receptor (FcRn). Fc receptors are a large family of proteins that bind to the Fc region of immunoglobulins (Igs) and function in Ig transport and effector functions. These receptors could function in enhancing the delivery of bNAbs in microbicide formulations or potentially serve as a mechanism of delivering HIV to target cells in tissues via transport of HIV-antibody complexes. Thus, this thesis assesses Fc receptor expression in human vaginal and endocervical organotypic cultures via microarray, quantitative RT-PCR, immunohistology and preliminary functional assays. Microarray results revealed significant expression of Fc receptor family genes in the epithelial cells of the lower female reproductive tract (FRT). The polymeric immunoglobulin receptor (pIgR), a well-characterized receptor that transports secretory IgA across mucosal epithelia, was abundantly expressed and hormonally regulated in epithelial cells of the vagina and endocervix. FcRn, a receptor originally characterized in the placenta and gut where it confers passive immunity from mother-to-child via bidirectional IgG transcytosis, was expressed in both tissue models. Moreover, several members of the novel FC receptor-like (FCRL) family were detected by microarray in both models. Immunohistological staining revealed pIgR protein in the endocervical mucosal epithelium, confirming current literature describing its expression in the FRT and role in local production of cervicovaginal secretions. FcRn protein expression was detected in the basal cell layer of the stratified squamous vaginal epithelium and in the columnar cells of the endocervix. Preliminary functional assays did not observe FcRn-specific transcytosis of human IgG across vaginal or endocervical epithelia by ELISA or immunohistology. VRCO1, a monoclonal antibody in development for application in microbicide formulation, crossed the epithelium, but was likely not transcytosed via FcRn because immunohistology revealed the presence of antibody between epithelial cells rather than the expected intracellular localization of IgG utilized in the FcRn mechanism. These preliminary findings indicate that Fc receptors, pIgR and Fc receptor-like proteins may play an important role in antibody-mediated immune responses in the FRT. Further research is require to determine whether FcRn functions in HIV-antibody complex-mediated HIV transmission or monoclonal antibody transcytosis

PREPARATION AND SOLID STATE CHARACTERIZATION OF NANOCRYSTALS FOR SOLUBILITY ENHANCEMENT OF QUETIAPINE FUMARATE

Objective: Quetiapine fumarate (QF) has poor water solubility and its oral bioavailability is observed to be 9% [1]. The objective of the present work is to prepare QF nanoparticles by wet media milling (WMM) and explore on the improvement in solubility with the aim of dose reduction and minimizing the side effects associated with its oral administration. Methods: Nanoparticle of QF were prepared by WMM using sodiumlaurylsulphate (SLS), polyvinylpyrrolidone (PVP), hydroxypropylcellulose (HPC) and Hydroxypropylmethylcellulose (HPMC) as surface stabilizers. The nanosuspensions obtained were characterized for its mean particle size (MPS), zeta-potential (ZP) and Polydispersity-index (PI). Further lyophilized nanoparticles were characterized for MPS, ZP, PI, saturation solubility (SS), X-ray diffraction (XRD), Differential Scanning Calorimetry (DSC), Infra-Red (IR) and Scanning electron Microscopy (SEM). Results: The MPS of the nanosuspensions obtained after 1 hour WMM and its lyophilized nanoparticlesis observed to be less than 1000 nm with PI ranges from 0.2 to 0.6 and negative ZP ranging from -30 to -36 mV. Lyophilized nanoparticles have shown significant improvement in SS when compared to plain drug. XRD and DSC have shown absence of the crystalline peaks and endotherm of QF respectively, indicating loss of its crystallinity. IR spectra showed no change in position of bands thus drug is compatible with surface stabilizers used. SEM photographs confirmed the formation of nanoparticles. Conclusion: From the above results it has been concluded that by subjecting QF to WMM, QF nanoparticles were obtained with significant improvement in saturation solubility and drug losing its crystalline nature when compared to plain drug

Active surface flows accelerate the coarsening of lipid membrane domains

Phase separation of multicomponent lipid membranes is characterized by the nucleation and coarsening of circular membrane domains that grow slowly in time as $\sim t^{1/3}$, following classical theories of coalescence and Ostwald ripening. In this work, we study the coarsening kinetics of phase-separating lipid membranes subjected to nonequilibrium forces and flows transmitted by motor-driven gliding actin filaments. We experimentally observe that the activity-induced surface flows trigger rapid coarsening of non-circular membrane domains that grow as $\sim t^{2/3}$, a 2$\times$ acceleration in the growth exponent compared to passive coalescence and Ostwald ripening. We analyze these results by developing analytical theories based on the Smoluchowski coagulation model and the phase field model to predict the domain growth in the presence of active flows. Our work demonstrates that active matter forces may be used to control the growth and morphology of membrane domains driven out of equilibrium.Comment: Main text is 5 pages with 3 figures. Supplemental materials attached include a supplemental appendix (includes supplemental movie legends, detailed methods, and derivations) and five supplemental movies (S1-S5

International Attitudes Toward the U.S., NATO and Russia in a Time of Crisis

This Pew Research Center analysis focuses on public opinion of the United States, Russia and NATO in 17 countries in North America, Europe, the Middle East and the Asia-Pacific region. The report draws on nationally representative surveys of 19,903 adults from Feb. 14 to May 11, 2022. All surveys were conducted over the phone with adults in Canada, Belgium, France, Germany, Greece, Italy, the Netherlands, Spain, Sweden, the United Kingdom, Japan, Malaysia, Singapore and South Korea. Surveys were conducted face to face in Poland and Israel and online in Australia.Data collection began a week prior to Russia's invasion of Ukraine in Canada, France, Germany, Italy, the UK and Japan. All other countries began fieldwork the same day as or shortly after the invasion. Due to the time it takes to translate, program and test questions on our international surveys, we prioritized gathering data at the start of this significant international event rather than delaying, or pausing, fieldwork to add questions specifically about the war or the actions taken by world leaders in response. Analysis focuses on ratings of Presidents Joe Biden and Vladimir Putin, the countries they lead and NATO as the war in Ukraine was unfolding. In this report, the data is discussed in the context of over a decade of cross-national trends.Views of Russia and NATO also include data from the United States. We surveyed 3,581 U.S. adults from March 21 to 27, 2022, after the start of the war in Ukraine. Everyone who took part in this survey is a member of the Center's American Trends Panel (ATP), an online survey panel that is recruited through national, random sampling of residential addresses. This way nearly all U.S. adults have a chance of selection. The survey is weighted to be representative of the U.S. adult population by gender, race, ethnicity, partisan affiliation, education and other categories

What Do Americans Know About International Affairs?

Americans know a great deal about certain global leaders and institutions. For example, nearly eight-in-ten U.S. adults can look at a photo of Kim Jong Un and correctly identify him as the leader of North Korea, and nearly two-thirds know that Boris Johnson is the current prime minister of the United Kingdom. A slim majority also know that Ukraine is not a member of the North Atlantic Treaty Organization (NATO).However, as a new Pew Research Center survey shows, Americans are less familiar with other topics. Despite the U.S. government labeling the events in Xinjiang, China, as genocide, only around one-in-five Americans are aware that it is the region in China with the most Muslims per capita. And only 41% can identify the flag of the second most populous country in the world, India

Effect of On-Ramp Demand on Capacity at Merge Bottleneck Locations

Past research acknowledged the impact of ramp vehicles on the occurrence of a breakdown event, but little has been done to quantify the effect of the ramp vehicles on the resulting bottleneck capacity. The objective of this research is to explore the relationship between ramp flow and capacity and to recommend capacity values for merge bottleneck locations. To explore the relationship between ramp flow and capacity, a capacity model has been developed using linear regression. The entire freeway network in Kansas City area was considered for the analysis. All locations which experienced “true breakdown”, breakdown because of merging operations and not due to downstream spillback, were selected for the analysis. Detector data at the six selected locations, were downloaded from KC Scout Portal from January 1, 2016 to June 30, 2016. Per lane and average speed, volume and occupancy data at 5-minute intervals were chosen for the analysis so as to detect breakdowns and find independent breakdown locations. Incident data and bad weather data were also collected for the same period and all the days with incidents and bad weather were removed from the analysis. For this research, free flow speed was defined as the average of flows when speeds were more than 50 miles per hour and the flows were less than 800 vehicles/hour. Breakdown was said to occur when speeds drop more than 25% of the free flow speed and the reduced speeds are maintained for at least 15 minutes, i.e., three 5-minute intervals (TRB, 2016). The breakdown capacities ranged from 3,900 to 8,500 vehicles per hour (veh/h) and when averaged across all the lanes, 1,150 to 2,200 vehicles per hour per lane (veh/h/ln). The upstream breakdown flows (demand) ranged from 3,400 to 8,400 veh/h and when averaged across all the lanes, 1,050-2,100 veh/h/ln. The ramp breakdown flows (ramp demand) ranged from 150 to 2,700 veh/h and when averaged across all the lanes, 150 to 1,500 veh/h/ln. Various variables such as freeway demand, ramp demand, free flow speed, number of lanes, ramp to freeway demand ratio, outer two-lane flow, shoulder lane flow, and remaining lane flow were considered for developing the model. Interactions between the variables were also considered. The final model was developed using 70% of the data, which were randomly selected, and the remaining 30% of the data was set aside for validation. A final model with an R2 value of 0.689 was developed. The high R2 value indicates that the developed model is a good predictor of capacity and this was also proven through the validation test for the developed model. The regression model was used to predict capacity values for different ramp demand and freeway demand. By observing the calculated capacity values it was concluded that the capacity was decreasing as the ramp demand, outer two lanes flows were increasing and the capacity per lane was decreasing as the number of lanes was decreasing, which is consistent with past literature (Lu & Elefeteriadou, 2013; Kondyli et al. 2016)

A Posthumanist Pragmatism: Rereading Tomboys

Gender has often dictated the roles and responsibilities that individuals are expected to fulfill. Societies in general still adhere to a strict gender binary system, and have largely been either intolerant of or, at minimum, uncomfortable with those who break from such a system. The tomboy figure has been the recipient of societal judgement for what has been interpreted to be a subversion of and deviance from traditional gender norms, and this has played out in a variety of ways. For instance, literary depictions of the tomboy—as the manifestations of the dominant cultural attitude—have captured both the aversion to as well as an evolving disposition toward non-feminine female characters. To trace and evaluate this trajectory, we utilize a framework provided by posthumanist theory, in conjunction with the pragmatic method. Important strains contained within posthumanism and pragmatism reject philosophical assumptions that there exists a single, true ontology, while promoting this-worldly notions concerning inclusion and diversity. By demonstrating how the tomboy has challenged presumptive ways of thinking, and continues to dispel preconceived notions and cultural expectations, we seek to show that the tomboy identity and disposition are to be celebrated for their authenticity and nonconformance, particularly as border-blurring and boundary-reducing, rather than deviating from or, for that matter, mirroring some purported true humanity. To accept such—that is, to hold any figure, tomboy or otherwise, as paradigmatic—would amount to a metaphysical endorsement for the knowability of a one and accurate experience, human or, even, otherwise. Alternatively, for us, the tomboy can serve as a valid model for how to undermine and help dismantle patriarchal and other prejudicial ideologies. Last, we attempt to show the increasing obsolescence of foundationalism as well as to ultimately offer the tomboy figure as a champion for continual self-enlargement, within a larger, posthumanist pragmatic process of self-creation

Attribute Based Secure Military Data Retrieval System for Decentralized Disruption Tolerant Networks

In this section, we provide a multi authority CP-ABE scheme for secure data retrieval in decentralized DTNs. Each local authority issues partial personalized and attribute key components to a user by performing secure 2PC protocol with the central authority. Each attribute key of a user can be updated individually and immediately. Thus, the scalability and security can be enhanced in the proposed scheme. Since the first CP-ABE scheme proposed by Bettencourt et al , dozens of CP-ABE schemes have been proposed. The subsequent CP-ABE schemes are mostly motivated by more rigorous security proof in the standard model. However, most of the schemes failed to achieve the expressiveness of the Bettencourt et al.’s scheme, which described an efficient system that was expressive in that it allowed an encrypt or to express an access predicate in terms of any monotonic formula over attributes. Therefore, in this section, we develop a variation of the CP-ABE algorithm partially based on (but not limited to) Bettencourt et al.’s construction in order to enhance the expressiveness of the access control policy instead of building a new CP-ABE scheme from scratc