Pre-treatment with systemic agents for advanced NSCLC elicits changes in the phenotype of autologous T cell therapy products.

The antitumor activity of adoptive T cell therapies (ACT) is highly dependent on the expansion, persistence, and continued activity of adoptively transferred cells. Clinical studies using ACTs have revealed that products that possess and maintain less differentiated phenotypes, including memory and precursor T cells, show increased antitumor efficacy and superior patient outcomes owing to their increased expansion, persistence, and ability to differentiate into effector progeny that elicit antitumor responses. Strategies that drive the differentiation into memory or precursor-type T cell subsets with high potential for persistence and self-renewal will enhance adoptively transferred T cell maintenance and promote durable antitumor efficacy. Because of the high costs associated with ACT manufacturing, ACTs are often only offered to patients after multiple rounds of systemic therapy. An essential factor to consider in producing autologous ACT medicinal products is the impact of the patient's initial T cell fitness and subtype composition, which will likely differ with age, disease history, and treatment with prior anti-cancer therapies. This study evaluated the impact of systemic anti-cancer therapy for non-small cell lung cancer treatment on the T cell phenotype of the patient at baseline and the quality and characteristics of the genetically modified autologous T cell therapy product after manufacturing.

Identification of a small molecule for enhancing lentiviral transduction of T cells.

Genetic modification of cells using viral vectors has shown huge therapeutic benefit in multiple diseases. However, inefficient transduction contributes to the high cost of these therapies. Several transduction-enhancing small molecules have previously been identified; however, some may be toxic to the cells or patient, otherwise alter cellular characteristics, or further increase manufacturing complexity. In this study, we aimed to identify molecules capable of enhancing lentiviral transduction of T cells from available small-molecule libraries. We conducted a high-throughput flow-cytometry-based screen of 27,892 compounds, which subsequently was narrowed down to six transduction-enhancing small molecules for further testing with two therapeutic lentiviral vectors used to manufacture GSK's clinical T cell therapy products. We demonstrate enhanced transduction without a negative impact on other product attributes. Furthermore, we present results of transcriptomic analysis, suggesting alteration of ribosome biogenesis, resulting in reduced interferon response, as a potential mechanism of action for the transduction-enhancing activity of the lead compound. Finally, we demonstrate the ability of the lead transduction enhancer to produce a comparable T cell product using a 3-fold reduction in vector volume in our clinical manufacturing process, resulting in a predicted 15% reduction in the overall cost of goods.

(I Think) You Are Pretty: a Behavior Analytic Conceptualization of Flirtation.

Much research in flirtation has been approached from a socio-cognitive perspective and has overemphasized subjective self-reports rather than overt behavior. Existing work pertinent to flirtation is reviewed here in addition to proposing a behavior-analytic perspective on the topic with a conception that includes both rule-governed and contingency-shaped behavior. Of particular interest within a verbal behavior conception of flirtation is the importance of autoclitics-features of a verbal response that affect the listener's reaction to the rest of the verbal response. Applications of a behavior analytic conception of flirtation and future directions relevant to research on interpersonal relationships are discussed.

Simulated Y-Site Compatibility of Vancomycin and Piperacillin-Tazobactam.

PURPOSE: To evaluate the physical compatibility of vancomycin with piperacillin-tazobactam during simulated Y-site administration. METHODS: Vancomycin and piperacillin-tazobactam were tested using 2 different diluents: 0.9% sodium chloride and 5% dextrose for injection. Vancomycin concentrations of 2, 5, and 10 mg/mL were tested using 0.9% sodium chloride and 4 and 8 mg/mL in 5% dextrose. Piperacillin-tazobactam was diluted to 16, 30, 40, 80, and 100 mg/mL, representing common concentrations used clinically in hospitals, and concentrations were tested in both 0.9% sodium chloride and 5% dextrose for injection. Medications were reconstituted under USP <797> aseptic technique. Combinations were tested in duplicate and reverse order with control solutions. Compatibility testing for Y-site included visual inspection, inspection with a high-intensity monodirectional light source (Tyndall beam), turbidimeter for turbidity evaluation, pH, and microscopic viewing. Testing occurred immediately after mixing, 15 minutes, 60 minutes, and 4 hours. If inconsistencies were observed between samples, testing was repeated to confirm results. Solutions were deemed incompatible if any one test failed and compatible if all tests were accepted. RESULTS: When dextrose 5% for injection was used as the diluent, vancomycin 4 mg/mL was Y-site compatible with piperacillin-tazobactam 16, 30, and 40 mg/mL and incompatible with 80 and 100 mg/mL. Vancomycin 8 mg/mL was incompatible with all tested concentrations of piperacillin-tazobactam. When 0.9% sodium chloride was used as the diluents, Y-site compatibility was found with vancomycin 2 and 5 mg/mL and all tested concentrations of piperacillin-tazobactam. Vancomycin 10 mg/mL was incompatible with piperacillin-tazobactam 40, 80, and 100 mg/mL. Incompatibilities formed a white precipitate immediately on mixing. CONCLUSION: Y-site incompatibility was greater for the tested concentrations of piperacillin-tazobactam and vancomycin when 5% dextrose was used as the diluent versus 0.9% sodium chloride. Y-site incompatibility was seen immediately in the form of a white precipitate on mixing.

Planar microdevices enhance transport of large molecular weight molecules across retinal pigment epithelial cells.

Large molecular weight drug delivery to the posterior eye is challenging due to cellular barriers that hinder drug transport. Understanding how to enhance transport across the retinal barrier is important for the design of new drug delivery systems. A novel mechanism to enhance drug transport is the use of geometric properties, which has not been extensively explored in the retina. Planar SU-8/Poly(ethyleneglycol)dimethacrylate microdevices were constructed using photolithography to deliver FITC dextran across an in vitro retinal model. The model consists of retinal pigment epithelial (RPE) cells grown to confluence on transwell inserts, which provides an environment to investigate the influence of geometry on paracellular and transcellular delivery of encapsulated large molecules. Planar microdevices enhanced transport of large molecular weight dextrans across different models of RPE in a size dependent fashion. Increased drug permeation across the RPE was observed with the addition of microdevices as compared to a traditional bolus of FITC dextran. This phenomena was initiated by a non-toxic interaction between the microdevices and the retinal tight junction proteins. Suggesting that increased drug transport occurs via a paracellular pathway. These experiments provide evidence to support the future use of planar unidirectional microdevices for delivery of biologics in ocular applications.

Planar microdevices for enhanced in vivo retention and oral bioavailability of poorly permeable drugs.

The development of novel oral drug delivery platforms for administering therapeutics in a safe and effective manner through the harsh gastrointestinal environment is of great importance. Here, the use of engineered thin planar poly(methyl methacrylate) (PMMA) microdevices is tested to enhance oral bioavailability of acyclovir, a poorly permeable drug. Acyclovir is loaded into the unidirectional drug releasing microdevice reservoirs using a drug entrapping photocross-linkable hydrogel matrix. An increase in acyclovir permeation across in vitro caco-2 monolayer is seen in the presence of microdevices as compared with acyclovir-entrapped hydrogels or free acyclovir solution. Cell proliferation studies show that microdevices are relatively nontoxic in nature for use in in vivo studies. Enhanced in vivo retention of microdevices is observed as their thin side walls experience minimal peristaltic shear stress as compared with spherical microparticles. Unidirectional acyclovir release and enhanced retention of microdevices achieve a 4.5-fold increase in bioavailability in vivo as compared with an oral gavage of acyclovir solution with the same drug mass. The enhanced oral bioavailability results suggest that thin, planar, bioadhesive, and unidirectional drug releasing microdevices will significantly improve the systemic and localized delivery of a broad range of oral therapeutics in the near future.

Contraceptive failure rates of etonogestrel subdermal implants in overweight and obese women.

OBJECTIVE: To estimate the contraceptive failure rates of the etonogestrel subdermal contraceptive implant in overweight and obese women and compare failure rates with women of normal weight and women using intrauterine devices (IUDs). METHODS: The Contraceptive CHOICE Project is a large prospective cohort study designed to promote the use of long-acting reversible contraceptive methods to reduce unintended pregnancies in the St Louis region. Participants are provided reversible contraception of their choice at no cost. We collected baseline height and weight of each participant. During each survey, participants were asked about missed menses and possible pregnancies. Any participant who suspected a pregnancy was asked to come in for urine pregnancy testing. Analysis includes the first 8,445 participants enrolled in CHOICE of which 1,168 chose the implant and 4,200 chose the IUD. Student's t test, χ test, and Kaplan-Meier survival curves were used to perform statistical analyses to estimate failure rates in overweight and obese women using the implant and IUDs. RESULTS: Of the women choosing the implant, 28% were overweight and 35% were obese. Of the women who chose an IUD, 27% were overweight and 35% were obese. The 3-year cumulative failure rates for implant and IUD users were less than one per 100 women-years and did not vary by body mass index. CONCLUSION: We found no decrease in the effectiveness of the implant in overweight or obese women. The implant may be offered as a first-line contraceptive method to any woman seeking a reversible and reliable birth control method.

A robust automated system elucidates mouse home cage behavioral structure.

Patterns of behavior exhibited by mice in their home cages reflect the function and interaction of numerous behavioral and physiological systems. Detailed assessment of these patterns thus has the potential to provide a powerful tool for understanding basic aspects of behavioral regulation and their perturbation by disease processes. However, the capacity to identify and examine these patterns in terms of their discrete levels of organization across diverse behaviors has been difficult to achieve and automate. Here, we describe an automated approach for the quantitative characterization of fundamental behavioral elements and their patterns in the freely behaving mouse. We demonstrate the utility of this approach by identifying unique features of home cage behavioral structure and changes in distinct levels of behavioral organization in mice with single gene mutations altering energy balance. The robust, automated, reproducible quantification of mouse home cage behavioral structure detailed here should have wide applicability for the study of mammalian physiology, behavior, and disease.

Fear appeals and binge drinking: a terror management theory perspective.

OBJECTIVES: The aim of the current research was to test the terror management theory-derived hypotheses that exposure to information about the mortality-related risks of binge drinking would make mortality salient (Study 1) and, hence, exacerbate willingness to binge drink amongst those who perceive this behaviour to benefit self-esteem (Study 2). STUDY 1: Participants (N=97) were allocated to one of five experimental conditions. Results confirmed that exposure to information about the mortality-related risks of binge drinking made mortality salient. STUDY 2: Participants (N=296) were allocated to one of three experimental conditions. Exposure to mortality-related information about the risks of binge drinking was found to result in greater willingness to binge drink among (i) binge drinkers and (ii) non-binge drinkers who perceived this behaviour to benefit self-esteem. There was no evidence, however, that exposure to such information influenced binge drinking over the following week. CONCLUSIONS: Research findings suggest that mortality-related health promotion campaigns might inadvertently make mortality salient, and hence precipitate the very behaviours which they aim to deter among some recipients.

Synergistic impairment of glucose homeostasis in ob/ob mice lacking functional serotonin 2C receptors.

To investigate how serotonin and leptin interact in the regulation of energy balance and glucose homeostasis, we generated a genetic mouse model, the OB2C mouse, which lacks functional serotonin 2C receptors and the adipocyte hormone leptin. The OB2C mice exhibited a dramatic diabetes phenotype, evidenced by a synergistic increase in serum glucose levels and water intake. The severity of the animals' diabetes phenotype would not have been predicted from the phenotypic characterization of mice bearing mutations of either the leptin (OB mutant mice) or the serotonin 2C receptor gene (2C mutant mice). The synergistic impairment in glucose homeostasis developed at an age when OB2C mice did not differ in body weight from OB mice, suggesting that this impairment was not an indirect consequence of increased adiposity. We also demonstrated that the improvement in glucose tolerance in wild-type mice treated with the serotonin releaser and reuptake inhibitor fenfluramine was blunted in 2C mutant mice. These pharmacological and genetic findings provide evidence that the serotonin 2C receptor has direct effects on glucose homeostasis.