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Clinical investigators often preprocess, process, and analyze their data without benefit of formally organized research centers to oversee data management. This article outlines a practical three-file structure to help guide these investigators track and document their data through processing and analyses. The proposed process can be implemented without additional training or specialized software. Thus, it is particularly well suited for research projects with small budgets or limited access to viable research/data coordinating centers. [ABSTRACT FROM AUTHOR]

Purpose: Estimating the sequential associations between educators' and children's talk during language learning interactions requires careful consideration of factors that may impact measurement stability and resultant inferences. This research note will describe a preliminary study that used generalizability theory to understand the contribution of two measurement conditions- occasions and raters -on estimates of sequential associations between educator talk and autistic preschooler talk in inclusive preschool classrooms., Method: We used an existing data set of four 15-min video-recorded occasions of educator-child interactions for 11 autistic preschoolers during free-play in their inclusive classroom. Two trained raters coded all videos for preschooler talk and type of educator talk (i.e., opportunities for expressive language [OELs], other talk). We conducted two generalizability studies on sequential association estimates for two interaction directions (i.e., preschooler talk following educator OEL and educator talk following preschooler talk). We conducted a series of decision studies to explore configurations of measurement conditions to optimize future investigations., Results: We had unstable estimates for both interaction directions in our current methodological approach, with raters accounting for minimal error and occasions accounting for considerable error. Future investigations would require at least six observation occasions for stable estimates of the sequential association between autistic preschooler talk following educator OEL that was stable after six occasions. More than 15 occasions were required for stable estimates of the association between educator talk following autistic preschooler talk., Conclusion: We will share recommendations and implications for future investigations to estimate educator and child talk sequential associations within preschool language interactions.

Multiple sclerosis (MS) is a chronic, progressive autoimmune disease modulated by autoantibodies that inflame and destroy the myelin sheath encasing neuronal axons, impairing proper axonal conduction and function. Glucagon-like peptide-1 (GLP-1) receptor agonists have been demonstrated to exert anti-inflammatory and neuroprotective effects, making these drugs particularly exciting prospects in the treatment of MS. While the exact mechanism remains unclear, GLP-1 receptor agonists may modulate inflammatory responses by targeting GLP-1 receptors present on immune cells such as macrophages, monocytes, and lymphocytes. In animal models, GLP-1 agonists have been shown to significantly delay the onset and severity of experimental autoimmune encephalopathy symptoms, as well as to increase nerve myelination and brain weight. In further experiments using animal models of nerve crush injury, specimens given GLP-1 agonists reported a significant increase in the rate and density of nerve regeneration compared to controls. Thus, GLP-1 agonists show promise as both prophylactic and symptomatic treatment for MS and may provide further utility in the treatment of other autoimmune, inflammatory, and neurodegenerative conditions., Competing Interests: Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Kaye et al.)

Organizations such as the National Clearinghouse on Autism Evidence and Practice have disseminated reports on evidence-based practices (EBPs) for individuals with autism to inform practice. However, some practitioners routinely employ unvalidated interventions more frequently than EBPs, signaling the presence of a research-to-practice gap. The current review investigated the degree to which single-case research studies with social communication outcomes for elementary-aged students with autism included details on variables that may impact the adoption and implementation of EBPs in practice. The results suggest limited reporting of variables that aid decision-making in the adoption and implementation of EBPs. Limited reporting can negatively impact uptake and fidelity and, therefore, student outcomes. Suggestions are made to improve researchers' reporting of the critical components of EBPs that will facilitate adoption and implementation by practitioners., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: There is little known about physical activity (PA) behaviors and its determinants in Cambodian adolescents, particularly with regard to the home and school settings. Therefore, we aimed to investigate these behaviors and determinants on their PA., Methods: The samples collected were from 168 high school students, aged 14-15 years. They were requested to complete the self-report PA questionnaire. Time spent in PA during weekday and weekend by school location and gender, and determinants on PA were analyzed. Differences in the mean values of the PA levels (in minutes) during weekday and the weekend between genders, and between school locations were analyzed using independent sample t tests. The percentage of students' perceptions on the determinants were calculated. Chi-squared test was used to compare the differences in prevalence of students' activities during their free time with respect to school location and gender., Results: The majority of the parents (86.9%-98.2%) demonstrated strong support for their children for academic work. Rural students spent an average time of moderate-to-vigorous PA greater than their counterparts from the urban areas during their weekend days (329.1 vs 239.2 min, respectively). Moderate to vigorous PA was likely to be higher during the weekends compared with weekdays among the boys (387.9 vs 361.4 min, respectively). Girls were more likely to spend their time on moderate to vigorous PA during the weekdays compared with the weekends (205.4 vs 180.5 min, respectively)., Conclusion: Consideration should be given to gender, school location, free time, and the environmental setting for the contextualization of more effective PA interventions with Cambodian youths.

B-cell lymphoma 6 (BCL6) is a transcriptional repressor and oncogenic driver of diffuse large B-cell lymphoma (DLBCL). Here, we report the optimization of our previously reported tricyclic quinolinone series for the inhibition of BCL6. We sought to improve the cellular potency and in vivo exposure of the non-degrading isomer, CCT373567 , of our recently published degrader, CCT373566 . The major limitation of our inhibitors was their high topological polar surface areas (TPSA), leading to increased efflux ratios. Reducing the molecular weight allowed us to remove polarity and decrease TPSA without considerably reducing solubility. Careful optimization of these properties, as guided by pharmacokinetic studies, led to the discovery of CCT374705 , a potent inhibitor of BCL6 with a good in vivo profile. Modest in vivo efficacy was achieved in a lymphoma xenograft mouse model after oral dosing.

Objective: In light of the current U.S. Food and Drug Administration (FDA) proposal to ban menthol cigarettes, this study updates trends in menthol cigarette use among adolescents age 13-18 years up to the year 2020. The study considers a potential role for the ban to reduce black/non-black disparities in menthol cigarette use, as well as a counterargument that a ban is not necessary because menthol use is already diminishing., Methods: Data are from annual, cross-sectional, nationally representative Monitoring the Future (MTF) surveys of 85 547 8th, 10th and 12th grade students surveyed between 2012 and 2020. Analyses include trends in past 30-day menthol and non-menthol cigarette smoking among the total adolescent population, as well as stratified by race/ethnicity., Results: Declines in adolescent menthol and non-menthol cigarette smoking continued through 2020 so that in 2018-2020 past 30-day prevalence for each was less than 1% for non-Hispanic black adolescents and less than 2.2% for non-black adolescents. For non-Hispanic black adolescents no smoking declines in mentholated or non-mentholated cigarette use from 2015-2017 to 2018-2020 were statistically significant, in part because prevalence levels approached a floor effect and had little room to fall further. Menthol levels were lower for non-Hispanic black versus all other adolescents in all study years., Conclusions: Continuing declines in adolescent menthol prevalence indicate that both menthol prevalence and also black/non-black disparities in its use are steadily decreasing. However, these decreases in adolescence will take decades to reach later ages through generational replacement. Efforts to accelerate menthol decreases will require new initiatives to increase cessation among adult menthol users., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. No commercial re-use. See rights and permissions. Published by BMJ.)

The transcriptional repressor BCL6 is an oncogenic driver found to be deregulated in lymphoid malignancies. Herein, we report the optimization of our previously reported benzimidazolone molecular glue-type degrader CCT369260 to CCT373566 , a highly potent probe suitable for sustained depletion of BCL6 in vivo . We observed a sharp degradation SAR, where subtle structural changes conveyed the ability to induce degradation of BCL6. CCT373566 showed modest in vivo efficacy in a lymphoma xenograft mouse model following oral dosing.

Check-in/Check-out (CICO) is a widely implemented evidence-based program for supporting students with at-risk levels of social and emotional behavior concerns. It is comprised of several core features described in the previous literature, including practice elements , which are the specific actions that are delivered directly to students, and implementation components, which are actions that support the implementation by adults. Practice elements and implementation components are both important to implementation but have been combined and conflated in descriptions of CICO implementation. Well-defined and differentiated practice elements could provide improved clarity in communicating implementation expectations to front-line implementers as well as support future research into essential active ingredients and measures of front-line intervention fidelity. The purpose of the present study was to distill, differentiate, and operationally define the student-facing practice elements of CICO. A panel of research experts and practice experts participated in a three-round modified e-Delphi process that led to the identification and operational definition of 19 discreet practice elements organized into five domains. Results are discussed in terms in implications for future development of measures of commitment and intervention fidelity, future research into active ingredients of CICO, and in terms of how well-defined practice elements can improve communication of implementation expectations for front-line implementers of CICO such as teachers., Supplementary Information: The online version contains supplementary material available at 10.1007/s12310-021-09495-x., Competing Interests: Conflicts of InterestThe authors declare that they have no conflict of interest., (© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2021, corrected publication 2022.)

Lay Abstract: Preschool special educators' are more likely to choose an educational practice to teach a young child with autism a social communication skill if they have positive beliefs about it. We asked preschool special educators to read a description of an autistic student and their social communication goal and imagine they were the student's teacher. We then asked them to pick one of five practices to teach the student. We also asked them questions to understand their attitudes about, confidence in their ability to use, and their perception of their coworkers' support of each practice. There are many research-based practices that a teacher could use to help children learn, and preschool teachers often make these decisions for their students. Teachers' beliefs varied in how supportive they were of each practice, and research shows people are more likely to do something that their beliefs support. In this study, they had more supportive beliefs and were more likely to use some practices, like naturalistic intervention, than other practices, like discrete trial teaching. By knowing this, researchers can help teachers use practices that their beliefs support and help change teachers' beliefs to be supportive of a practice they may need to use.

We describe the optimization of modestly active starting points to potent inhibitors of BCL6 by growing into a subpocket, which was occupied by a network of five stably bound water molecules. Identifying potent inhibitors required not only forming new interactions in the subpocket but also perturbing the water network in a productive, potency-increasing fashion while controlling the physicochemical properties. We achieved this goal in a sequential manner by systematically probing the pocket and the water network, ultimately achieving a 100-fold improvement of activity. The most potent compounds displaced three of the five initial water molecules and formed hydrogen bonds with the remaining two. Compound 25 showed a promising profile for a lead compound with submicromolar inhibition of BCL6 in cells and satisfactory pharmacokinetic (PK) properties. Our work highlights the importance of finding productive ways to perturb existing water networks when growing into solvent-filled protein pockets.

The objective of the present study was to determine whether there is an association between pornography use and reported intimate partner violence (IPV) perpetration among a sample of soldiers in the US Army. The study was a secondary analysis of cross-sectional data collected from soldiers at a military installation in 2018 (n = 9,052). IPV was defined as any self-reported perpetration of physical, sexual, or psychological abuse of an intimate partner. Multivariable negative binomial regressions were used to assess the association between pornography use and any lifetime perpetration of IPV, controlling for gender, age group, race/ethnicity, relationship status, educational status, military rank, hazardous drinking, depression, stimulant use, depressant use, and post-traumatic stress disorder. Of the population analyzed, 41% of soldiers reported any pornography use per week, and 9.6% reported perpetrating any form of IPV. Soldiers who reported pornography use had between a 1.72- and 3.56-fold greater likelihood of reporting any lifetime perpetration of IPV, controlling for covariates. Given the prevalence and detrimental effects of IPV, longitudinal studies should be designed to further understand predictors of IPV in military populations., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

Purpose A myriad features can impact the nature, frequency, and length of adult-child interactions important for language learning. Empirical investigations of language learning opportunities for young children with autism spectrum disorder (ASD) provide limited generalizable insight, with inferences more constrained to the sample than is often considered. The aim of this study was to explore a multidimensional understanding of reliability and define optimal measurement procedures for a measurement approach used to examine the language of preschool educators interacting with children with ASD. Method We employed the logic of Generalizability Theory to differentiate sources of error for two measurement facets, occasion and observer . We video-recorded four 15-min occasions of educator-child interactions for 11 participants with ASD during free-play in their respective inclusive preschool classrooms. Two trained observers coded all videos for six educator language variables: open-ended questions/statements, choice questions, yes/no questions, imitation prompts, statements, and other talk. Results The generalizability studies illustrated that, across all variables measured, observer accounted for little to no error. Occasion, however, accounted for much of the error for all language variables. To determine the number of occasions needed to achieve stable estimates of the variables, we manipulated occasion in the decision study. Five to more than 15 occasions were needed to achieve stability in educator language variables. Conclusion To advance our understanding of the language learning environments of preschool classrooms that serve children with ASD, researchers must understand how aspects of the measurement design in those environments, such as occasion, impact the inferences they make.

Deregulation of the transcriptional repressor BCL6 enables tumorigenesis of germinal center B-cells, and hence BCL6 has been proposed as a therapeutic target for the treatment of diffuse large B-cell lymphoma (DLBCL). Herein we report the discovery of a series of benzimidazolone inhibitors of the protein-protein interaction between BCL6 and its co-repressors. A subset of these inhibitors were found to cause rapid degradation of BCL6, and optimization of pharmacokinetic properties led to the discovery of 5-((5-chloro-2-((3 R ,5 S )-4,4-difluoro-3,5-dimethylpiperidin-1-yl)pyrimidin-4-yl)amino)-3-(3-hydroxy-3-methylbutyl)-1-methyl-1,3-dihydro-2 H -benzo[ d ]imidazol-2-one (CCT369260), which reduces BCL6 levels in a lymphoma xenograft mouse model following oral dosing.

Tumor-associated macrophages (TAMs) are heterogeneous and can adopt a spectrum of activation states between pro-inflammatory and pro-tumorigenic in response to the microenvironment. We have previously shown that TTI-621, a soluble SIRPαFc fusion protein that blocks the CD47 "do-not-eat" signal, promotes tumor cell phagocytosis by IFN-γ-primed macrophages. To assess the impact of CD47 blockade on diverse types of macrophages that are found within the tumor microenvironment, six different polarized human macrophage subsets (M(-), M(IFN-γ), M(IFN-γ+LPS), M(IL-4), M(HAGG+IL-1β), M(IL-10 + TGFβ)) with distinct cell surface markers and cytokine profiles were generated. Blockade of CD47 using TTI-621 significantly increased phagocytosis of lymphoma cells by all macrophage subsets, with M(IFN-γ), M(IFN-γ+LPS) and M(IL-10 + TGFβ) macrophages having the highest phagocytic response. TTI-621-mediated phagocytosis involves macrophage expression of both the low- and high-affinity Fcγ receptors II (CD32) and I (CD64), respectively. Moreover, macrophages with lower phagocytic capabilities (M(-), M(IL-4), M(HAGG+IL-1β)) could readily be re-polarized into highly phagocytic macrophages using various cytokines or TLR agonists. In line with the in vitro study, we further demonstrate that TTI-621 can trigger phagocytosis of tumor cells by diverse subsets of isolated mouse TAMs ex vivo. These data suggest that TTI-621 may be efficacious in triggering the destruction of cancer cells by a diverse population of TAMs found in vivo and support possible combination approaches to augment the activity of CD47 blockade.

Purpose: The ubiquitously expressed transmembrane glycoprotein CD47 delivers an anti-phagocytic (do not eat) signal by binding signal-regulatory protein α (SIRPα) on macrophages. CD47 is overexpressed in cancer cells and its expression is associated with poor clinical outcomes. TTI-621 (SIRPαFc) is a fully human recombinant fusion protein that blocks the CD47-SIRPα axis by binding to human CD47 and enhancing phagocytosis of malignant cells. Blockade of this inhibitory axis using TTI-621 has emerged as a promising therapeutic strategy to promote tumor cell eradication. Experimental Design: The ability of TTI-621 to promote macrophage-mediated phagocytosis of human tumor cells was assessed using both confocal microscopy and flow cytometry. In vivo antitumor efficacy was evaluated in xenograft and syngeneic models and the role of the Fc region in antitumor activity was evaluated using SIRPαFc constructs with different Fc tails. Results: TTI-621 enhanced macrophage-mediated phagocytosis of both hematologic and solid tumor cells, while sparing normal cells. In vivo , TTI-621 effectively controlled the growth of aggressive AML and B lymphoma xenografts and was efficacious in a syngeneic B lymphoma model. The IgG1 Fc tail of TTI-621 plays a critical role in its antitumor activity, presumably by engaging activating Fcγ receptors on macrophages. Finally, TTI-621 exhibits minimal binding to human erythrocytes, thereby differentiating it from CD47 blocking antibodies. Conclusions: These data indicate that TTI-621 is active across a broad range of human tumors. These results further establish CD47 as a critical regulator of innate immune surveillance and form the basis for clinical development of TTI-621 in multiple oncology indications. Clin Cancer Res; 23(4); 1068-79. ©2016 AACR ., (©2016 American Association for Cancer Research.)

Aim: Delay in instituting neuroprotective measures after cardiac arrest increases death and decreases neuronal recovery. Current hypothermia methods are slow, ineffective, unreliable, or highly invasive. We report the feasibility of rapid hypothermia induction in swine through augmented heat extraction from the lungs., Methods: Twenty-four domestic crossbred pigs (weight, 50-55kg) were ventilated with room air. Intraparenchymal brain temperature and core temperatures from pulmonary artery, lower esophagus, bladder, rectum, nasopharynx, and tympanum were recorded. In eight animals, ventilation was switched to cooled helium-oxygen mixture (heliox) and perfluorocarbon (PFC) aerosol and continued for 90min or until target brain temperature of 32°C was reached. Eight animals received body-surface cooling with water-circulating blankets; eight control animals continued to be ventilated with room air., Results: Brain and core temperatures declined rapidly with cooled heliox-PFC ventilation. The brain reached target temperature within the study period (mean [SD], 66 [7.6]min) in only the transpulmonary cooling group. Cardiopulmonary functions and poststudy histopathological examination of the lungs were normal., Conclusion: Transpulmonary cooling is novel, rapid, minimally invasive, and an effective technique to induce therapeutic hypothermia. High thermal conductivity of helium and vaporization of PFC produces rapid cooling of alveolar gases. The thinness and large surface area of alveolar membrane facilitate rapid cooling of the pulmonary circulation. Because of differences in thermogenesis, blood flow, insulation, and exposure to the external environment, the brain cools at a different rate than other organs. Transpulmonary hypothermia was significantly faster than body surface cooling in reaching target brain temperature., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Standard cancer treatments trigger immune responses that may influence tumor control. The nature of these responses varies depending on the tumor and the treatment modality. We previously reported that radiation and androgen-deprivation therapy (ADT) induce tumor-associated autoantibody responses in prostate cancer patients. This follow-up analysis was conducted to assess the relationship between autoantibody responses and clinical outcome. Patients with non-metastatic prostate cancer received external beam radiation therapy (EBRT) plus neoadjuvant and concurrent androgen deprivation. Treatment-induced autoantibodies were detected in almost a third of patients receiving combinatorial ADT and EBRT. Unexpectedly, patients that developed autoantibody responses to tumor antigens had a significantly lower 5-year biochemical failure-free survival (BFFS) than patients that did not develop an autoantibody response. Thus, tumor-reactive autoantibodies may be associated with increased risk of biochemical failure and immunomodulation to prevent autoantibody development may improve BFFS for select, high-risk prostate cancer patients receiving both ADT and EBRT.

Introduction: Standard precautions are disease transmission prevention strategies recommended by both the World Health Organization (WHO) and by the Centers for Disease Control and Prevention (CDC). Emergency medical services (EMS) personnel are expected to utilize standard precautions., Methods: This was a prospective observational study of the use of standard precautions by EMS providers arriving at a large urban emergency department (ED). Research assistants (RAs) observed EMS crews throughout their arrival and delivery of patients and recorded data related to the use of gloves, hand hygiene, and equipment disinfection., Results: A total of 423 EMS deliveries were observed, allowing for observation of 899 EMS providers. Only 512 (56.9%) EMS providers arrived wearing gloves. Hand washing was observed in 250 (27.8%) of providers. Reusable equipment disinfection was noted in only 31.6% of opportunities. The most commonly disinfected item was the stretcher (55%)., Conclusion: EMS provider compliance with standard precautions and equipment disinfection recommendations is suboptimal. Strategies must be developed to improve EMS provider compliance with internationally recognized infection control guidelines. Key words: Emergency medical services, hand washing, hygiene, disinfection, disease prevention.

A better understanding of the interplay between the tumor environment, the immune system, and hemostatic apparatus is essential to effectively improve cancer treatment. Histidine-rich glycoprotein (HRG) is an abundant plasma protein with a wide array of functions. HRG has the ability to bind multiple ligands thereby modulating immunity, cell adhesion, angiogenesis, and thrombosis. Many of these functions are involved in tumor progression and antitumor response. We outline current data on HRG as an important potential player and as a potential future target for cancer therapy.

Purpose: To provide rationale for using phosphoinositide 3-kinase (PI3K) and/or mitogen-activated protein kinase (MAPK) pathway inhibitors to treat rhabdomyosarcomas, a major cause of pediatric and adolescent cancer deaths., Experimental Design: The prevalence of PI3K/MAPK pathway activation in rhabdomyosarcoma clinical samples was assessed using immunohistochemistry. Compensatory signaling and cross-talk between PI3K/MAPK pathways was determined in rhabdomyosarcoma cell lines following p110α short hairpin RNA-mediated depletion. Pharmacologic inhibition of reprogrammed signaling in stable p110α knockdown lines was used to determine the target-inhibition profile inducing maximal growth inhibition. The in vitro and in vivo efficacy of inhibitors of TORC1/2 (AZD8055), MEK (AZD6244), and P13K/mTOR (NVP-BEZ235) was evaluated alone and in pairwise combinations., Results: PI3K pathway activation was seen in 82.5% rhabdomyosarcomas with coactivated MAPK in 36% and 46% of alveolar and embryonal subtypes, respectively. p110α knockdown in cell lines over the short and long term was associated with compensatory expression of other p110 isoforms, activation of the MAPK pathway, and cross-talk to reactivate the PI3K pathway. Combinations of PI3K pathway and MAP-ERK kinase (MEK) inhibitors synergistically inhibited cell growth in vitro. Treatment of RD cells with AZD8055 plus AZD6244 blocked reciprocal pathway activation, as evidenced by reduced AKT/ERK/S6 phosphorylation. In vivo, the synergistic effect on growth and changes in pharmacodynamic biomarkers was recapitulated using the AZD8055/AZD6244 combination but not NVP-BEZ235/AZD6244. Pharmacokinetic analysis provided evidence of drug-drug interaction with both combinations., Conclusions: Dual PI3K/MAPK pathway activation and compensatory signaling in both rhabdomyosarcoma subtypes predict a lack of clinical efficacy for single agents targeting either pathway, supporting a therapeutic strategy combining a TORC1/2 with a MEK inhibitor.

Background: Biomarker validation remains one of the most challenging constraints to the development of new diagnostic assays. To facilitate biomarker validation, we previously developed a chromatography-free stable isotope standards and capture by antipeptide antibodies (SISCAPA)-MALDI assay allowing rapid, high-throughput quantification of protein analytes in large sample sets. Here we applied this assay to the measurement of a surrogate proteotypic peptide from protein C inhibitor (PCI) in sera from patients with prostate cancer., Methods: A 2-plex SISCAPA-MALDI assay for quantification of proteotypic peptides from PCI and soluble transferrin receptor (sTfR) was used to measure these peptides in 159 trypsin-digested sera collected from 51 patients with prostate cancer. These patients had been treated with radiation with or without neoadjuvant androgen deprivation., Results: Patients who experienced biochemical recurrence of prostate cancer showed decreased serum concentrations of the PCI peptide analyte within 18 months of treatment. The PCI peptide concentrations remained increased in the sera of patients who did not experience cancer recurrence. Prostate-specific antigen concentrations had no predictive value during the same time period., Conclusions: The high-throughput, liquid chromatography-free SISCAPA-MALDI assay is capable of rapid quantification of proteotypic PCI and sTfR peptide analytes in complex serum samples. Decreased serum concentrations of the PCI peptide were found to be related to recurrence of prostate cancer in patients treated with radiation with or without hormone therapy. However, a larger cohort of patients will be required for unequivocal validation of the PCI peptide as a biomarker for clinical use.

The pleiotropic cytokine TGF-β has been implicated in the regulation of numerous aspects of the immune response, including naïve T cell homeostasis. Previous studies found that impairing TGF-β responsiveness (through expression of a dominant-negative TGF-β RII [DNRII] transgene) leads to accumulation of memory phenotype CD8 T cells, and it was proposed that this resulted from enhanced IL-15 sensitivity. Here we show naïve DNRII CD8 T cells exhibit enhanced lymphopenia-driven proliferation and generation of "homeostatic" memory cells. However, this enhanced response occurred in the absence of IL-15 and, unexpectedly, even in the combined absence of IL-7 and IL-15, which were thought essential for CD8 T cell homeostatic expansion. DNRII transgenic CD8 T cells still require access to self Class I MHC for homeostatic proliferation, arguing against generalized dysregulation of homeostatic cues. These findings suggest TGF-β responsiveness is critical for enforcing sensitivity to homeostatic cytokines that limit maintenance and composition of the CD8 T cell pool. (154 words).

Objective: This study measured health literacy in a population of teens in treatment for asthma or diabetes and tested the association between health literacy and willingness to use online health resources., Materials and Methods: About 180 patients aged 13-18 years treated for asthma or diabetes in specialty care clinics completed assessments of demographic characteristics, health literacy, and Internet access and use. Teens were provided a resource page listing selected publically available health-related Web sites and asked about perceived ease of use, perceived usefulness, and intent to use the listed Web sites. The relationship between demographic characteristics, health literacy, and online health information use was tested using chi-squared or Fisher's exact test. Predictors of intent to use resource page Web sites were assessed using bivariate and multivariate ordinal logistic regression., Results: About 92% of participants had adequate health literacy. Over 50% of participants had previously searched online for health information. Older age was the only significant predictor of health information search. Most teens (79%) reported intent to use at least one Web site from the resource page at least occasionally within the next 3 months. Higher health literacy (odds ratio [OR]=6.24, p<0.01) and stronger perceived usefulness (OR=1.74, p=0.01) were associated with greater intent for regular use, after controlling for demographic and Internet access variables., Conclusions: Teens with lower health literacy searched online for health information as often as peers with higher literacy, but were less likely to express the intent to use recommended sites. Belief in the usefulness of a Web site is the strongest attitudinal predictor of intended future use.

Objective: Publically available health Web sites may be able to provide support for teens with chronic disease in their transition to autonomy. This study examines teen use of health Web sites and identifies barriers and promoters of use., Materials and Methods: One hundred eighty (n=180) teens, aged 13-18, were recruited from asthma and diabetes specialty clinics and given a resource sheet listing selected publically available Web sites. Web sites were categorized as general health, teen health, disease specific (asthma or diabetes), and disease management. One hundred twenty-nine (n=129) participants completed a 3-month follow-up semistructured telephone interview that assessed subsequent use of resource sheet Web sites and collected verbatim comments regarding use., Results: Sixty percent visited at least one Web site from the resource sheet. General health (38.8%) and teen health Web sites (38.0%) were most likely to be visited. Reasons for not visiting any site were categorized as lack of time, lack of interest, lack of Internet access, and lack of medical need. Teen health sites had the highest intention for continued use. Comments associated with willingness to continue use were categorized into the following themes: usefulness, ease, medical need, teen-specific features, and frequency of updates. Neither health literacy nor demographic factors were significantly associated with use., Conclusions: Making a first Web site visit generally leads to a desire for continued use, but teens will not make the first effort of visiting a Web site unless they are engaged and interested in their own health management. Teens are most willing to become regular users of Web sites that they perceive to be useful and to be targeted toward them.

Background: Host T-cell responses are associated with favorable outcomes in epithelial ovarian cancer (EOC), but it remains unclear how best to promote these responses in patients. Toward this goal, we evaluated a panel of clinically relevant cytokines for the ability to enhance multiple T-cell effector functions (polyfunctionality) in the native tumor environment., Methodology/principal Findings: Experiments were performed with resident CD8+ and CD4+ T cells in bulk ascites cell preparations from high-grade serous EOC patients. T cells were stimulated with α-CD3 in the presence of 100% autologous ascites fluid with or without exogenous IL-2, IL-12, IL-18 or IL-21, alone or in combination. T-cell proliferation (Ki-67) and function (IFN-γ, TNF-α, IL-2, CCL4, and CD107a expression) were assessed by multi-parameter flow cytometry. In parallel, 27 cytokines were measured in culture supernatants. While ascites fluid had variable effects on CD8+ and CD4+ T-cell proliferation, it inhibited T-cell function in most patient samples, with CD107a, IFN-γ, and CCL4 showing the greatest inhibition. This was accompanied by reduced levels of IL-1β, IL-1ra, IL-9, IL-17, G-CSF, GM-CSF, Mip-1α, PDGF-bb, and bFGF in culture supernatants. T-cell proliferation was enhanced by exogenous IL-2, but other T-cell functions were largely unaffected by single cytokines. The combination of IL-2 with cytokines engaging complementary signaling pathways, in particular IL-12 and IL-18, enhanced expression of IFN-γ, TNF-α, and CCL4 in all patient samples by promoting polyfunctional T-cell responses. Despite this, other functional parameters generally remained inhibited., Conclusions/significance: The EOC ascites environment disrupts multiple T-cell functions, and exogenous cytokines engaging diverse signaling pathways only partially reverse these effects. Our results may explain the limited efficacy of cytokine therapies for EOC to date. Full restoration of T-cell function will require activation of signaling pathways beyond those engaged by IL-2, IL-12, IL-18, and IL-21.

Upon transfer into T cell-deficient hosts, naive CD8(+) T cells typically undergo lymphopenia-induced proliferation (LIP, also called homeostatic proliferation) and develop the phenotypic and functional characteristics of memory CD8(+) T cells. However, the capacity of T cells with self-peptide/MHC specificity to respond in this way has not been intensively studied. We examined pmel-1 TCR transgenic CD8(+) T cells that are specific for an epitope from gp100, a protein expressed by melanoma cells and normal melanocytes. Despite their self-specificity, naive pmel-1 cells were inefficient at LIP in typical lymphopenic hosts. In CD132 (common gamma-chain)-deficient hosts, pmel-1 CD8(+) T cells underwent extensive proliferation, but, surprisingly, the majority of these cells retained certain naive phenotypic traits (CD44(low), CD122(low)) rather than acquiring the expected central-memory phenotype. Following LIP, pmel-1 T cells acquired the capacity to control B16F10 tumor growth, but only in common gamma-chain-deficient host mice. Together, these data suggest that LIP does not always favor expansion of self-specific CD8 T cells and that sustained extensive lymphopenia is required for such cells to exhibit tumor control.

Memory T cells exhibit superior responses to pathogens and tumors compared with their naive counterparts. Memory is typically generated via an immune response to a foreign antigen, but functional memory T cells can also be produced from naive cells by homeostatic mechanisms. Using a recently developed method, we studied CD8 T cells, which are specific for model (ovalbumin) and viral (HSV, vaccinia) antigens, in unimmunized mice and found a subpopulation bearing markers of memory cells. Based on their phenotypic markers and by their presence in germ-free mice, these preexisting memory-like CD44(hi) CD8 T cells are likely to arise via physiological homeostatic proliferation rather than a response to environmental microbes. These antigen-inexperienced memory phenotype CD8 T cells display several functions that distinguish them from their CD44(lo) counterparts, including a rapid initiation of proliferation after T cell stimulation and rapid IFN-gamma production after exposure to proinflammatory cytokines. Collectively, these data indicate that the unprimed antigen-specific CD8 T cell repertoire contains antigen-inexperienced cells that display phenotypic and functional traits of memory cells.

It is now apparent that naïve peripheral T cells are a dynamic population where active processes prevent inappropriate activation while supporting survival. The process of thymic education makes naïve peripheral T cells dependent on interactions with self-MHC for survival. However, as these signals can potentially result in inappropriate activation, various non-redundant, intrinsic negative regulatory molecules including Tob, Nfatc2, and Smad3 actively enforce T cell quiescence. Interactions among these pathways are only now coming to light and may include positive or negative crosstalk. In the case of positive crosstalk, self-MHC initiated signals and intrinsic negative regulatory factors may cooperate to dampen T cell activation and sustain peripheral tolerance in a binary fashion (on-off). In the case of negative crosstalk, self-MHC signals may promote survival through partial activation while intrinsic negative regulatory factors act as rheostats to restrain cell cycle entry and prevent T cells from crossing a threshold that would break tolerance.

In past decades waste was left to the lowest level of staff to dispose of at the lowest cost. Today, business managers in leading companies consider waste to be part of their responsibility. There is money to be made by eliminating waste problems. Leading companies are moving to the top of the waste disposal hierarchy and eliminating waste for themselves and their customers.

Eyeblink classical conditioning is a relatively simple form of associative learning that has become an invaluable tool in our understanding of the neural mechanisms of learning. When studying rabbits in this paradigm, we observed a dramatic modification of learning rate by conducting training during episodes of either hippocampal theta or hippocampal non-theta activity as determined by on-line slow-wave spectral analysis. Specifically, if animals were given trials only when a computer analysis verified a predominance of slow-wave oscillations at theta frequencies (3-8 Hz), they learned in half as many trials as animals trained during non-theta hippocampal activity (58 vs. 115). This finding provides important evidence from awake, behaving animals that supports recent advances in our knowledge of (i) brain sites and neurobiological mechanisms of learning and memory, specifically hippocampus and theta oscillations, (ii) the biological plausibility of current models of hippocampal function that posit important roles for oscillatory potentials, and (iii) the design of interfaces between biological and cybernetic (electronic) systems that can optimize cognitive processes and performance.

Ca(2+)/calmodulin-dependent protein kinase II (CaMK-II) isozyme variability is the result of alternative usage of variable domain sequences. Isozyme expression is cell type-specific to transduce the appropriate Ca(2+) signals. We have determined the subcellular targeting domain of delta(E) CaMK-II, an isozyme that induces neurite outgrowth, and of a structurally similar isozyme, gamma(C) CaMK-II, which does not induce neurite outgrowth. delta(E) CaMK-II co-localizes with filamentous actin in the perinuclear region and in cellular extensions. In contrast, gamma(C) CaMK-II is uniformly cytosolic. Constitutively active delta(E) CaMK-II induces F-actin-rich extensions, thereby supporting a functional role for its localization. C-terminal constructs, which lack central variable domain sequences, can oligomerize and localize like full-length delta(E) and gamma(C) CaMK-II. Central variable domains themselves are monomeric and have no targeting capability. The C-terminal 95 residues of delta CaMK-II also has no targeting capability but can efficiently oligomerize. These findings define a targeting domain for gamma and delta CaMK-IIs that is in between the central variable and association domains. This domain is responsible for the subcellular targeting differences between gamma and delta CaMK-IIs.

A prospective study of 43 cotton-top tamarins, from infancy to 6 to 17 months of age, was conducted to determine the epidemiology of Campylobacter spp. infection. Nine infants followed for one year in an isolation unit, where attendants wore protective clothing, did not become infected. In the main facility where 32 of 34 animals had repeated infections with C. coli, 6% of the infections developed initially in incubators, 66% in the nursery room, and 28% after transfer to the main colony. Fifteen of these tamarins also were infected with C. jejuni. Twenty percent of the infections developed initially in the nursery room and 80% in the colony. Polyacrylamide gel electrophoresis analysis of C. jejuni cultures revealed multiple reinfections with different strains. Both types of infections were most prevalent between 3 and 9 months of age. Campylobacterjejuni infection developed most frequently between April and June and C. coli infection developed between October and December. In the nursery, diarrhea developed most frequently at times when there was no infection with Campylobacter spp. Forty percent of animals with diarrhea in the nursery had C. coli and none had C. jejuni, whereas, in the colony, 49% had C. jejuni and 11% had C. coli infections. There was no association between these infections and diet or idiopathic colitis.

Ca(2+)/calmodulin-dependent protein kinase II (CaMK-II) has been linked to the induction of differentiation in preneuronal cells. In these cells, delta isozymes represent the majority of CaMK-IIs expressed and are activated by differentiation stimuli. To determine whether delta CaMK-IIs are causative or coincident with in vitro differentiation, we overexpressed wild-type, constitutively active, and C-terminal domains of delta and gamma CaMK-II isozymes in mouse P19 and NIH/3T3 cells using high-efficiency transfections. At 1-2 days after transfection, only constitutively active delta CaMK-II isozymes induced branched cellular extensions in both cell types. In P19 cells, retinoic acid induced neurite extensions after 3-4 days; these extensions were coincident with a fourfold increase in endogenous CaMK-II activity. Extensions induced by both retinoic acid and delta CaMK-IIs contained class III beta-tubulin in a discontinuous or beaded pattern. C-terminal CaMK-II constructs disrupted the ability of endogenous CaMK-II to autophosphorylate and blocked retinoic acid-induced differentiation. delta CaMK-II was found along extensions, whereas gamma CaMK-II exhibited a more diffuse, cytosolic localization. These data not only support an extranuclear role for CaMK-II in promoting neurite outgrowth, but also demonstrate CaMK-II isozyme specificity in these early steps of neuronal differentiation.