The Positive Effect of the Rational Addiction Prevention Program (RAPP) on Adolescents with High Risk for Drug Consumption.

The aim of this longitudinal study was to determine the effect of an original prevention program (RAPP) on the behavioral and cognitive characteristics of adolescents with high risk for substance consumption. Seventy-six Mexican adolescents 12-15 years old (38 with high risk (HR) and 38 with low risk (LR)) were selected. RAPP was applied for 3 months. Resilience, social skills, attitudes towards substance use, ability to delay a reward, and inhibitory control were assessed in these adolescents, before and after the RAPP intervention. Both groups improved their scores; however, HR achieved greater changes than LR. Findings suggest that HR have behavioral characteristics that can be considered as risk factors for substance consumption (low levels of resilience, low social skills, little family support, positive attitudes towards substance use). RAPP proved to be an effective program for preventing these risk factors for substance use in adolescents.

Synthesis and Biological Evaluation of a Carbamate-Containing Tubulysin Antibody-Drug Conjugate.

Antibody-drug conjugates (ADCs) use antibodies to deliver cytotoxic payloads directly into tumor cells via specifically binding to the target cell surface antigens. ADCs can enhance the anti-tumor effects of antibodies, and increase the delivery of cytotoxic payloads to cancer cells with a better therapeutic index. An ADC was prepared with a potent carbamate-containing tubulysin analogue attached to an anti-mesothelin antibody via a Cit-Val dipeptide linker. An aniline functionality in the tubulysin analogue was created to provide a site of linker attachment via an amide bond that would be stable in systemic circulation. Upon ADC internalization into antigen-positive cancer cells, the Cit-Val dipeptide linker was cleaved by lysosomal proteases, and the drug was released inside the tumor cells. The naturally occurring acetate of tubulysin was modified to a carbamate to reduce acetate hydrolysis of the ADC in circulation and to increase the hydrophilicity of the drug. The ADC bearing the monoclonal anti-mesothelin antibody and the carbamate-containing tubulysin was highly potent and immunologically specific to H226 human lung carcinoma cells in vitro, and efficacious at well-tolerated doses in a mesothelin-positive OVCAR3 ovarian cancer xenograft mouse model.

High-Throughput Platform to Identify Antibody Conjugation Sites from Antibody-Drug Conjugate Libraries.

Antibody-drug conjugates (ADCs) are a therapeutic modality that traditionally enable the targeted delivery of highly potent cytotoxic agents to specific cells such as tumor cells. More recently, antibodies have been used to deliver molecules such as antibiotics, antigens, and adjuvants to bacteria or specific immune cell subsets. Site-directed mutagenesis of proteins permits more precise control over the site and stoichiometry of their conjugation, giving rise to homogeneous chemically defined ADCs. Identification of favorable sites for conjugation in antibodies is essential as reaction efficiency and product stability are influenced by the tertiary structure of immunoglobulin G (IgG). Current methods to evaluate potential conjugation sites are time-consuming and labor intensive, involving multistep processes for individually produced reactions. Here, we describe a highly efficient method for identification of conjugatable genetic variants by analyzing pooled ADC libraries using mass spectrometry. This approach provides a versatile platform to rapidly uncover new conjugation sites for site-specific ADCs.

Parent perspectives on food allergy management and safety during the COVID-19 pandemic.

Background: U.S. national emergency was declared in mid-March 2020 due to the coronavirus disease 2019 (COVID-19) pandemic. Subsequently, a period of stay-at-home orders, regulatory changes, evolving medical recommendations, and food supply chain disruptions occurred. There is little published research on how such changes affected food allergy management for children with this diagnosis. Objective: The study goal was to identify parent perspectives with regard to if and/or how pandemic-related regulatory changes and evolving medical recommendations have affected food allergy management. Methods: A survey was distributed to parents of children with food allergy. An electronic Internet forms survey link was available for completion during July 2020. Data were presented as descriptive statistics, cleaned, and coded into a spreadsheet before analysis. Frequencies and percentage were calculated to describe participants' characteristics and responses. Results: Of 377 responses, 359 met inclusion criteria. Concerns about COVID-19 exposure were expressed in 65.7% about accessing an emergency department and 73.6% had school reentry concerns; 66% had not discussed recommended anaphylaxis management algorithm changes with a provider; 85.8% had not discussed the temporary U.S. Food and Drug Administration food labeling policy with a provider. Most (62%) reported shortages of preferred safe food brands. 62% spent more time cooking safe foods from scratch. With regard to the recommendation by the U.S. Centers for Disease Control and Prevention (CDC) for classroom dining, 57.7% planned to request modifications. With regard to the CDC's recommendation to use inhalers versus nebulizers, 37.7% had not discussed the topic with a provider. Ninety-two written comments were analyzed and grouped into seven themes. Conclusion: New pandemic-related regulations, food supply chain disruptions, and evolving medical recommendations resulted in intensified burdens for respondents, including the increased time needed to complete food allergy management and school reentry concerns. Study results can inform clinical team members (e.g., physicians, nurses, dieticians) of effects that pandemic-related changes may have on this patient population, with subsequent consideration of patient-specific screening, education, and shared decision-making with regard to risk mitigation needs.

Cognitive variability-A marker for incident MCI and AD: An analysis for the Alzheimer's Disease Neuroimaging Initiative.

INTRODUCTION: The potential of intra-individual cognitive variability (IICV) to predict incident mild cognitive impairment (MCI) or Alzheimer's disease (AD) was examined and compared to well-established neuroimaging and genetic predictors. METHODS: IICV was estimated using four neuropsychological measures for n = 1324 Alzheimer's Disease Neuroimaging Initiative (ADNI) participants who were cognitively healthy or diagnosed with MCI at baseline. IICV was used to predict time to incident MCI or AD, and compared to hippocampal volume loss and APOE ε4 status via survival analysis. RESULTS: In survival analyses, controlling for age, education, baseline diagonosis, and APOE ε4 status, likelihood ratio tests indicate that IICV is associated with time to cognitive status change in the full sample (P < .0001), and when the sample was restricted to individuals with MCI at baseline (P < .0001). DISCUSSION: These findings suggest IICV may be a low-cost, noninvasive alternative to traditional AD biomarkers.

Initial report of the osteogenesis imperfecta adult natural history initiative.

BACKGROUND: A better understanding of the natural history of osteogenesis imperfecta (OI) in adulthood should improve health care for patients with this rare condition. METHODS: The Osteogenesis Imperfecta Foundation established the Adult Natural History Initiative (ANHI) in 2010 to give voice to the health concerns of the adult OI community and to begin to address existing knowledge gaps for this condition. Using a web-based platform, 959 adults with self-reported OI, representing a wide range of self-reported disease severity, reported symptoms and health conditions, estimated the impact of these concerns on present and future health-related quality of life (QoL) and completed a Patient-Reported Outcomes Measurement Information System (PROMIS®) survey of health issues. RESULTS: Adults with OI report lower general physical health status (p < .0001), exhibit a higher prevalence of auditory (58% of sample versus 2-16% of normalized population) and musculoskeletal (64% of sample versus 1-3% of normalized population) concerns than the general population, but report generally similar mental health status. Musculoskeletal, auditory, pulmonary, endocrine, and gastrointestinal issues are particular future health-related QoL concerns for these adults. Numerous other statistically significant differences exist among adults with OI as well as between adults with OI and the referent PROMIS® population, but the clinical significance of these differences is uncertain. CONCLUSIONS: Adults with OI report lower general health status but are otherwise more similar to the general population than might have been expected. While reassuring, further analysis of the extensive OI-ANHI databank should help identify areas of unique clinical concern and for future research. The OI-ANHI survey experience supports an internet-based strategy for successful patient-centered outcomes research in rare disease populations.

The PALS prevention program and its long-term impact on student intentions to use alcohol, tobacco, and marijuana.

A unique Alcohol, Tobacco, and Other Drug (ATOD) prevention program called PALS (Prevention through Alternative Learning Styles) was implemented with middle school students with the goal of enhancing student knowledge of the harmful effects of ATOD, promoting the use of refusal skills and reducing intentions to use ATOD in the future. Intentions to use were measured at four points: pre-PALS, post-PALS, and at 1-year and 2-year follow-ups. Student survey responses were then matched and compared across the four time periods. This article reports on the long-term effectiveness of PALS on student intentions to use ATOD in high school. When follow-up surveys of PALS students were compared to students not exposed to PALS (comparison group), the PALS students had significantly lower intentions to use alcohol and tobacco, providing evidence that the PALS intervention did have a long-term impact on intentions to use these substances.

An ecological approach to addressing HIV/AIDS in the African American community.

The disproportionate impact of HIV/AIDS on African Americans is a significant public health challenge. The complex constellation of individual, social, and environmental factors influencing transmission, require ecological solutions that recognize these multiple levels of influence and actively involve communities. This article describes the formation of a community-based coalition and highlights three initiatives it has undertaken in the areas of mobile HIV testing, HIV education, and faith-based work to improve HIV services for African Americans.

Evaluating the prevention through alternative learning styles program.

This article reports on the evaluation of a two-year alcohol, tobacco and other drug (ATOD) intervention, the Prevention through Alternative Learning Styles (PALS) program, targeting both teachers and middle-school students. Teachers are taught to recognize students' unique learning styles in the context of the ATOD curriculum and adapt the ATOD messages to these learning styles. The student curriculum consists of 5 topic areas with two lessons per topic area. Student goals include enhancing students' knowledge of the effects of ATOD, promoting students' use of refusal skills and decreasing students' intentions to use ATOD. The program was implemented in school dis-tricts in the greater Dayton Ohio area. Support was found for the intervention's overall effectiveness in both years, with statistically significant improvements demonstrated by the students who participated in the PALS program. Students had an increase in their knowledge of ATOD topic areas and a decrease in their intentions to use ATOD.

Efficacy of contact lens disinfecting solutions against Staphylococcus aureus and Pseudomonas aeruginosa.

PURPOSE: To evaluate the disinfection properties of multipurpose contact lens disinfection solutions, based on the International Organization for Standardization (ISO) 14729 guidelines. METHODS: ReNu with MoistureLoc Multi-Purpose Solution, OPTI-FREE Express with Aldox Multi-Purpose Solution, Betadine 5% sterile ophthalmic preparation solution (povidone iodine), and 0.9% normal saline solution were inoculated with strains of Staphylococcus aureus and Pseudomonas aeruginosa. Surviving bacteria were quantified at specified times. RESULTS: ReNu with MoistureLoc, OPTI-FREE Express, and 5% ophthalmic povidone iodine were effective in achieving a 5-log reduction in bacterial count. Additionally, all three products maintained their effectivity at 72 hours. However, ReNu with MoistureLoc and povidone iodine resulted in the greatest reduction in bacterial colonization. CONCLUSIONS: ReNu with MoistureLoc, OPTI-FREE Express, and 5% ophthalmic povidone iodine meet the ISO 14729 guidelines for standalone contact lens solutions. However, ReNu with MoistureLoc and 5% ophthalmic povidone iodine are most efficient in reducing and maintaining low bacterial count for a period of 72 hours.

IL-6 levels predict disease variant and extent of organ involvement in patients with mastocytosis.

Mastocytosis is often associated with organ involvement and hematological disorders. Patients may also exhibit elevated levels of plasma IL-6. To gain insight into the relevance of this observation, we correlated plasma levels of IL-6 and soluble IL-6 receptor (sIL-6R) with multiple disease parameters in 29 patients with mastocytosis. Mean plasma IL-6 levels were elevated in patients compared to healthy controls (P < 0.0001). Disease category significantly correlated with plasma IL-6 levels, as did severity of bone marrow pathology, organomegaly, and extent of skin involvement. In plasma, there was a positive correlation of IL-6 to total tryptase, alkaline phosphatase, IgM, white blood cell count, prothrombin time, partial thromboplastin time, and neutrophil numbers. There was an inverse correlation to hemoglobin. sIL-6R levels were not elevated. These observations demonstrate that IL-6 is a useful surrogate marker of severity of hematologic disease and suggest that IL-6 contributes to pathology.

Alterations of plasma lipids in mice via adenoviral-mediated hepatic overexpression of human ABCA1.

ATP binding cassette transporter A1 (ABCA1) is a widely expressed lipid transporter essential for the generation of HDL. ABCA1 is particularly abundant in the liver, suggesting that the liver may play a major role in HDL homeostasis. To determine how hepatic ABCA1 affects plasma HDL cholesterol levels, we treated mice with an adenovirus (Ad)-expressing human ABCA1 under the control of the cytomegalovirus promoter. Treated mice showed a dose-dependent increase in hepatic ABCA1 protein, ranging from 1.2-fold to 8.3-fold using doses from 5 x 108 to 1.5 x 109 pfu, with maximal expression observed on Day 3 posttreatment. A selective increase in HDL cholesterol occurred at Day 3 in mice treated with 5 x 108 pfu Ad-ABCA1, but higher doses did not further elevate HDL cholesterol levels. In contrast, total cholesterol, triglycerides, phospholipids, non-HDL cholesterol, and apolipoprotein B levels all increased in a dose-dependent manner, suggesting that excessive overexpression of hepatic ABCA1 in the absence of its normal regulatory sequences altered total lipid homeostasis. At comparable expression levels, bacterial artificial chromosome transgenic mice, which express ABCA1 under the control of its endogenous regulatory sequences, showed a greater and more specific increase in HDL cholesterol than Ad-ABCA1-treated mice. Our results suggest that appropriate regulation of ABCA1 is critical for a selective increase in HDL cholesterol levels.

Assessment of the extent of cutaneous involvement in children and adults with mastocytosis: relationship to symptomatology, tryptase levels, and bone marrow pathology.

BACKGROUND: Cutaneous involvement occurs in most patients with systemic mastocytosis. OBJECTIVE: We sought to determine whether the extent of cutaneous involvement is predictive of systemic disease. METHODS: In a prospective survey of 48 adults and 19 children, the extent and density of cutaneous lesions were compared with patient history, symptoms, internal organ involvement, serum total mast cell tryptase level, and bone marrow pathology. RESULTS: Cutaneous lesions in children were of a greater mean and maximum diameter, but similar in extent and density compared with lesions in adults. In adults with skin lesions, the extent of lesions correlated to disease duration. Adults with extensive cutaneous disease experienced more pruritus and flushing. Fatigue, splenomegaly, and hepatomegaly were more frequent in adults without cutaneous involvement; and in those with a greater density of lesions and disease duration. Increased tryptase levels were found in children and adults with systemic disease and correlated to skin lesion density and bone marrow pathology. CONCLUSION: An examination of the extent and density of cutaneous lesions in adults helps identify those with more extensive extracutaneous disease and, thus, requiring a more thorough evaluation.

Protein kinase A site-specific phosphorylation regulates ATP-binding cassette A1 (ABCA1)-mediated phospholipid efflux.

ATP-binding cassette A1 (ABCA1) is a key mediator of cholesterol and phospholipid efflux to apolipoprotein particles. We show that ABCA1 is a constitutively phosphorylated protein in both RAW macrophages and in a human embryonic kidney cell line expressing ABCA1. Furthermore, we demonstrate that phosphorylation of ABCA1 is mediated by protein kinase A (PKA) or a PKA-like kinase in vivo. Through site-directed mutagenesis studies of consensus PKA phosphorylation sites and in vitro PKA kinase assays, we show that Ser-1042 and Ser-2054, located in the nucleotide binding domains of ABCA1, are major phosphorylation sites for PKA. ApoA-I-dependent phospholipid efflux was decreased significantly by mutation of Ser-2054 alone and Ser-1042/Ser-2054 but was not significantly impaired with Ser-1042 alone. The mechanism by which ABCA1 phosphorylation affected ApoA-I-dependent phospholipid efflux did not involve either alterations in ApoA-I binding or changes in ABCA1 protein stability. These studies demonstrate a novel serine (Ser-2054) on the ABCA1 protein crucial for PKA phosphorylation and for regulation of ABCA1 transporter activity.

Regression of urticaria pigmentosa in adult patients with systemic mastocytosis: correlation with clinical patterns of disease.

OBJECTIVE: To determine clinical correlates of urticaria pigmentosa (UP) regression in adult patients with systemic mastocytosis (SM). DESIGN: Cohort study of the natural history of mastocytosis. SETTING: National Institutes of Health Clinical Center. PATIENTS: In a study of adult patients referred to the National Institutes of Health after 1980 and observed for a minimum of 10 years, 12 of 106 adult patients experienced clearance or fading of UP. MAIN OUTCOME MEASURES: Data from each patient's history and results of physical examination, laboratory evaluation, and organ biopsy at presentation to the National Institutes of Health were compared with findings at the patient's most recent visit. RESULTS: In the patients in whom clearance of (n = 5) or a decrease in skin lesions (n = 7) was noted, UP had persisted from 4 to 34 years (median, 17 years). Older age was a prognostic feature for regression of UP. Despite improvement of UP, the 2 patients with SM with an associated hematologic disorder experienced a deterioration in clinical condition. In the 10 patients with indolent SM, severity and frequency of symptoms decreased as the UP regressed. However, bone marrow changes consistent with SM remained. CONCLUSIONS: Urticaria pigmentosa regresses in approximately 10% of the older patients who have SM. In patients with an associated hematologic disorder such as myelodysplasia, this regression may be accompanied by disease progression. In contrast, regression of UP in patients with indolent SM parallels a decrease in disease intensity, although bone marrow findings of indolent SM remain.

Human cytomegalovirus US7, US8, US9, and US10 are cytoplasmic glycoproteins, not found at cell surfaces, and US9 does not mediate cell-to-cell spread.

Human cytomegalovirus (HCMV) expresses a large number of membrane proteins with unknown functions. One class of these membrane proteins apparently acts to allow HCMV to escape detection by the immune system. The best characterized of these are the glycoproteins encoded within the US2 to US11 region of the HCMV genome that mediate resistance to CD8(+) and CD4(+) T cells. US2, US3, US6, and US11 block various aspects of the major histocompatibility complex (MHC) class I and class II antigen presentation pathways, functioning in cytoplasmic membranes to cause retention, degradation, or mislocalization of MHC proteins. Distantly homologous genes in this region, US7, US8, US9, and US10, are not well characterized. Here, we report expression of the glycoproteins encoded by US7 to US10 by using replication-defective adenovirus (Ad) vectors. US7, US9, and US10 remained sensitive to endoglycosidase H and were exclusively or largely present in the endoplasmic reticulum (ER) as determined by confocal microscopy. US8 reached the Golgi apparatus and trans-Golgi network and was more quickly degraded. Previous studies suggested that US9 could localize to cell junctions and mediate cell-to-cell spread in ARPE-19 retinal epithelial cells. We found no evidence of US9 at cell junctions of HEC-1A epithelial cells. HCMV recombinants lacking US9 produced smaller plaques on ARPE-19 cell monolayers but also exhibited defects in virus replication compared with wild-type HCMV in these cells. Other HCMV recombinants constructed in a similar fashion that were able to express US9 also produced small plaques and some of these exhibited defects in production of infectious progeny in ARPE-19 cells. Thus, there was no correlation between defects in cell-to-cell spread (plaque size) and loss of expression of US9, and it is possible that US9(-) mutants produce smaller plaques because they produce fewer progeny. Together, our results do not support the hypothesis that US9 plays a direct role in HCMV cell-to-cell spread.

Levels of mast-cell growth factors in plasma and in suction skin blister fluid in adults with mastocytosis: correlation with dermal mast-cell numbers and mast-cell tryptase.

BACKGROUND: Mast-cell accumulation has been observed in the skin and other organs of patients with systemic indolent mastocytosis (SM). The basis for this pathologic increase is not fully understood. OBJECTIVE: We sought to determine levels of mast-cell growth factors in the skin and plasma of patients with SM, patients with atopic dermatitis (AD), and healthy individuals and to correlate these levels to dermal mast-cell numbers and levels of mast-cell tryptase. METHODS: Skin suction blister fluid and plasma levels of stem-cell factor, IL-3, IL-4, IL-6, vascular endothelial growth factor, and total mast-cell tryptase were analyzed by means of ELISA. The number of mast cells was determined in a biopsy section taken from adjacent skin. RESULTS: Mast-cell numbers in the dermis were higher in patients with SM compared with numbers in patients with AD (P <.001) or in healthy control subjects (P <.0001) and correlated with tryptase levels in both skin blister fluid (P <.0001) and plasma (P <.0001). Stem-cell factor and vascular endothelial growth factor levels in the skin blister fluid and plasma of patients with SM were not significantly different from those in patients with AD or healthy control subjects. IL-3 and IL-4 levels were below the limit of detection. IL-6 levels were significantly increased in the plasma of patients with SM compared with in plasma of patients with AD (P <.002) and healthy control subjects (P <.0001) and correlated with plasma tryptase levels (P <.05) and dermal mast-cell numbers (P <.02). CONCLUSION: Because elevated levels of IL-6 could contribute to the fever, fatigue, and osteoporosis observed in patients with SM and because IL-6 is antiapoptotic for mast cells, IL-6 could potentiate the biologic consequences of this disease.

Suppression of IL-4 Production in Murine Lymphocytes by Orally Effective Prostaglandin E(1) Analogs.

Prostaglandins (PGs) regulate a wide variety of immunologic processes. We studied the activity of PGE(1), and two orally effective PGE(1) methyl ester analogs, misoprostol (MP) and enisoprost (EP), as inhibitors of interleukin-4 (IL-4) production in stimulated murine splenic T cells in vitro. Effective suppresion of IL-4 production was seen with all three agents with KI(50)s of 65 nM for MP and 20 nM for EP. IL-4 is primarily a product of type 2 T helper cells (Th2 cells) and plays an important role in regulating the biosynthesis of immunoglobulins, especially IgE. These in vitro studies, taken together with the recent data of others on modulation of IgE responses by EP and MP in immunized mice in vivo, suggest a possible role for these PGE(1) analogs in the treatment of IgE-mediated diseases in humans.