"God Loves All Equally and Infinitely": A Mixed Methods Analysis of LGBTQ+ Ally Development Within Religious/Spiritual Contexts.

Within the United States, conservative theological perspectives have exerted influential sociocultural influences toward LGBTQ+ communities, with profound implications toward ally development. Ally development can present its challenges, especially for those in non-affirming religious and/or spiritual (R/S) contexts. We employed a mixed methods approach, utilizing both Enhanced Critical Incident Technique and chi-square tests of independence, focusing on the self-identified incidents that influenced the internal negotiation process of 315 LGBTQ+ allies within R/S contexts. Participants described critical R/S incidents that impacted their ally development, and these incidents fell into four main categories: (1) the influence of religious organizations, leaderships, and congregants, (2) the impact of meaningful relationships, (3) the experience of spiritual or divine shifting events, and (4) meaning making. Additionally, participants identified wish list items represented by four main themes: (1) fostering ally development, (2) taking action, (3) nothing should have changed, and (4) wishing for support broadly. Chi-square tests of independence revealed that R/S incidences were related to specific wish list items.

Laser cooling of ions in a neutral plasma.

Laser cooling of a neutral plasma is a challenging task because of the high temperatures typically associated with the plasma state. By using an ultracold neutral plasma created by photoionization of an ultracold atomic gas, we avoid this obstacle and demonstrate laser cooling of ions in a neutral plasma. After 135 microseconds of cooling, we observed a reduction in ion temperature by up to a factor of four, with the temperature reaching as low as 50(4) millikelvin. This pushes laboratory studies of neutral plasmas deeper into the strongly coupled regime, beyond the limits of validity of current kinetic theories for calculating transport properties. The same optical forces also retard the plasma expansion, opening avenues for neutral-plasma confinement and manipulation.

Assembly of a three-dimensional multitype bronchiole coculture model using magnetic levitation.

A longstanding goal in biomedical research has been to create organotypic cocultures that faithfully represent native tissue environments. There is presently great interest in representative culture models of the lung, which is a particularly challenging tissue to recreate in vitro. This study used magnetic levitation in conjunction with magnetic nanoparticles as a means of creating an organized three-dimensional (3D) coculture of the bronchiole that sequentially layers cells in a manner similar to native tissue architecture. The 3D coculture model was assembled from four human cell types in the bronchiole: endothelial cells, smooth muscle cells (SMCs), fibroblasts, and epithelial cells (EpiCs). This study represents the first effort to combine these particular cell types into an organized bronchiole coculture. These cell layers were first cultured in 3D by magnetic levitation, and then manipulated into contact with a custom-made magnetic pen, and again cultured for 48 h. Hematoxylin and eosin staining of the resulting coculture showed four distinct layers within the 3D coculture. Immunohistochemistry confirmed the phenotype of each of the four cell types and showed organized extracellular matrix formation, particularly, with collagen type I. Positive stains for CD31, von Willebrand factor, smooth muscle α-actin, vimentin, and fibronectin demonstrate the maintenance of the phenotype for endothelial cells, SMCs, and fibroblasts. Positive stains for mucin-5AC, cytokeratin, and E-cadherin after 7 days with and without 1% fetal bovine serum showed that EpiCs maintained the phenotype and function. This study validates magnetic levitation as a method for the rapid creation of organized 3D cocultures that maintain the phenotype and induce extracellular matrix formation.

A high throughput microelectroporation device to introduce a chimeric antigen receptor to redirect the specificity of human T cells.

It has been demonstrated that a chimeric antigen receptor (CAR) can directly recognize the CD19 molecule expressed on the cell surface of B-cell malignancies independent of major histocompatibility complex (MHC). Although T-cell therapy of tumors using CD19-specific CAR is promising, this approach relies on using expression vectors that stably integrate the CAR into T-cell chromosomes. To circumvent the potential genotoxicity that may occur from expressing integrating transgenes, we have expressed the CD19-specific CAR transgene from mRNA using a high throughput microelectroporation device. This research was accomplished using a microelectroporator to achieve efficient and high throughput non-viral gene transfer of in vitro transcribed CAR mRNA into human T cells that had been numerically expanded ex vivo. Electro-transfer of mRNA avoids the potential genotoxicity associated with vector and transgene integration and the high throughput capacity overcomes the expected transient CAR expression, as repeated rounds of electroporation can replace T cells that have lost transgene expression. We fabricated and tested a high throughput microelectroporator that can electroporate a stream of 2 x 10(8) primary T cells within 10 min. After electroporation, up to 80% of the passaged T cells expressed the CD19-specific CAR. Video time-lapse microscopy (VTLM) demonstrated the redirected effector function of the genetically manipulated T cells to specifically lyse CD19+ tumor cells. Our biomedical microdevice, in which T cells are transiently and safely modified to be tumor-specific and then can be re-infused, offers a method for redirecting T-cell specificity, that has implications for the development of adoptive immunotherapy.

Ultracold neutral plasmas.

Ultracold neutral plasmas occupy an exotic regime of plasma physics in which electrons form a swarming, neutralizing background for ions that sluggishly move in a correlated manner. Strong interactions between the charged particles give rise to surprising dynamics such as oscillations of the average kinetic energy during equilibration and extremely fast recombination. Such phenomena offer stimulating and challenging problems for computational scientists, and the physics can be applied to other environments, such as the interior of gas giant planets and plasmas created by short-pulse laser irradiation of solid, liquid, and cluster targets.