Addressing Health Disparities with School-Based Outreach: the Health Career Academy Program.

Pipeline programs address health disparities by promoting academic achievement and entry of low-income ethnic and racial minority youth into healthcare fields. The Health Career Academy (HCA) is a 3-year pipeline program for high school students from low-income, ethnic, and racial minority communities. Health professional students serve as program mentors. The HCA has been implemented in nine US sites, with partnerships between 17 health professional schools and 17 high schools. A total of 386 10th grade students and 95 11th grade students enrolled as participants in the 2015-2016 HCA program. In post-participation surveys, 10th grade students reported that the HCA helped them learn about different healthcare career options, plan for how to reach career goals, and understand how healthcare workers care for patients. Eleventh grade participants noted the program made them aware of the importance of public health and taught them about medical conditions, self-care, and safety. Eighty-six percent of 10th graders and 71% of 11th graders reported that they are considering healthcare careers. Students' favorite aspects of the HCA included the following: time with mentors, learning about science and health, team collaboration and hands-on experiences, field trips, and team presentations. Teachers noted the following as most important in the program: interaction with mentors and healthcare professionals, learning broadly applicable skills, stimulation of interest in health-related careers, presentation skills, and creating optimism about furthering education. The HCA is well received by participants and can be replicated successfully at multiple sites nationally. By providing mentorship, increasing exposure to health professionals and health careers, offering high-level science and health curriculum, and fostering collaboration and presentation skills, the HCA has potential to increase interest in health professions among racial and ethnic minority youth from low-income communities.

Identification of Antibody and Small Molecule Antagonists of Ferroportin-Hepcidin Interaction.

The iron exporter ferroportin and its ligand, the hormone hepcidin, control fluxes of stored and recycled iron for use in a variety of essential biochemical processes. Inflammatory disorders and malignancies are often associated with high hepcidin levels, leading to ferroportin down-regulation, iron sequestration in tissue macrophages and subsequent anemia. The objective of this research was to develop reagents to characterize the expression of ferroportin, the interaction between ferroportin and hepcidin, as well as to identify novel ferroportin antagonists capable of maintaining iron export in the presence of hepcidin. Development of investigative tools that enabled cell-based screening assays is described in detail, including specific and sensitive monoclonal antibodies that detect endogenously-expressed human and mouse ferroportin and fluorescently-labeled chemically-synthesized human hepcidin. Large and small molecule antagonists inhibiting hepcidin-mediated ferroportin internalization were identified, and unique insights into the requirements for interaction between these two key iron homeostasis molecules are provided.

Bispecific T cell engager (BiTE®) antibody constructs can mediate bystander tumor cell killing.

For targets that are homogenously expressed, such as CD19 on cells of the B lymphocyte lineage, immunotherapies can be highly effective. Targeting CD19 with blinatumomab, a CD19/CD3 bispecific antibody construct (BiTE®), or with chimeric antigen receptor T cells (CAR-T) has shown great promise for treating certain CD19-positive hematological malignancies. In contrast, solid tumors with heterogeneous expression of the tumor-associated antigen (TAA) may present a challenge for targeted therapies. To prevent escape of TAA-negative cancer cells, immunotherapies with a local bystander effect would be beneficial. As a model to investigate BiTE®-mediated bystander killing in the solid tumor setting, we used epidermal growth factor receptor (EGFR) as a target. We measured lysis of EGFR-negative populations in vitro and in vivo when co-cultured with EGFR-positive cells, human T cells and an EGFR/CD3 BiTE® antibody construct. Bystander EGFR-negative cells were efficiently lysed by BiTE®-activated T cells only when proximal to EGFR-positive cells. Our mechanistic analysis suggests that cytokines released by BiTE®-activated T-cells induced upregulation of ICAM-1 and FAS on EGFR-negative bystander cells, contributing to T cell-induced bystander cell lysis.

Medical Student Volunteerism Addresses Patients' Social Needs: A Novel Approach to Patient-Centered Care.

BACKGROUND: Healthcare providers must be equipped to recognize and address patients' psychosocial needs to improve overall health outcomes. To give future healthcare providers the tools and training necessary to identify and address psychosocial issues, Lankenau Medical Center in partnership with the Philadelphia College of Osteopathic Medicine designed the Medical Student Advocate (MSA) program. METHODS: The MSA program places volunteer second-year osteopathic medical students in care coordination teams at Lankenau Medical Associates, a primary care practice serving a diverse patient population in the Philadelphia, PA, region. As active members of the team, MSAs are referred high-risk patients who have resource needs such as food, employment, child care, and transportation. MSAs work collaboratively with patients and the multidisciplinary team to address patients' nonmedical needs. RESULTS: From August 2013 to August 2015, 31 osteopathic medical students volunteered for the MSA program and served 369 patients with 720 identified needs. Faculty and participating medical students report that the MSA program provided an enhanced understanding of the holistic nature of patient care and a comprehensive view of patient needs. CONCLUSION: The MSA program provides students with a unique educational opportunity that encompasses early exposure to patient interaction, social determinants of health, population health, and interdisciplinary collaboration. Students develop skills to help them build patient relationships, understand the psychosocial factors shaping health outcomes, and engage with other healthcare professionals. This work in the preclinical years provides students with the knowledge to help them perform more effectively in the changing healthcare environment.

Development and initial evaluation of electronic Children's International Mucositis Evaluation Scale (eChIMES) for children with cancer.

PURPOSE: We previously developed a pediatric-specific measure of oral mucositis named the Children's International Mucositis Evaluation Scale (ChIMES). Availability as an electronic version may improve self-report response rates. The objectives were to develop an electronic version of ChIMES (eChIMES) and to determine whether the instrument is easy to use, understandable, and suitable for measuring mucositis among children and adolescents with cancer. METHODS: Development of eChIMES was on an iPad; the initial version was piloted with ten children to refine instructions for use and presentation. A crosssectional study then was conducted and included English-speaking children and adolescents 8-18 years of age receiving active treatment for cancer. Participants were shown eCHIMES and were asked to complete it. Questions elicited whether they found eChIMES easy or difficult to use, easy or difficult to understand, and suitable (a good way) for children with cancer to monitor mucositis. Outcomes were rated using five-point ordinal scales. RESULTS: Following the development and initial refinement of eChIMES, 40 children were enrolled. Median age was 12.4 (range, 8.0 to 17.8) years. The instrument was found to be easy or very easy to use and understood by 40 (100 %) and 38 (95 %) participants, respectively. The application was considered suitable or very suitable for measuring mucositis by 37 (92 %). CONCLUSIONS: We found that eChIMES was easy to use, understandable, and suitable for monitoring mucositis among children with cancer. Incorporation into clinical trials may improve the ability to compare and evaluate interventions for mucositis.

Molecular mechanism of hepcidin-mediated ferroportin internalization requires ferroportin lysines, not tyrosines or JAK-STAT.

Ferroportin is the primary means of cellular iron efflux and a key component of iron metabolism. Hepcidin regulates Fpn activity by inducing its internalization and degradation. The mechanism of internalization is reported to require JAK2 activation, phosphorylation of Fpn tyrosine residues 302 and 303, and initiation of transcription through STAT3 phosphorylation. These findings suggest Fpn may be a target for therapeutic intervention through JAK2 modulation. To evaluate the proposed mechanism, Fpn internalization was assessed using several techniques combined with reagents that specifically recognized cell-surface Fpn. In vitro results demonstrated that Hepc-induced Fpn internalization did not require JAK2 or phosphorylation of Fpn residues 302 and 303, nor did it induce JAK-STAT signaling. In vivo, inhibition of JAK2 had no effect on Hepc-induced hypoferremia. However, internalization was delayed by mutation of two Fpn lysine residues that may be targets of ubiquitination.

Preclinical evaluation of AMG 900, a novel potent and highly selective pan-aurora kinase inhibitor with activity in taxane-resistant tumor cell lines.

In mammalian cells, the aurora kinases (aurora-A, -B, and -C) play essential roles in regulating cell division. The expression of aurora-A and -B is elevated in a variety of human cancers and is associated with high proliferation rates and poor prognosis, making them attractive targets for anticancer therapy. AMG 900 is an orally bioavailable, potent, and highly selective pan-aurora kinase inhibitor that is active in taxane-resistant tumor cell lines. In tumor cells, AMG 900 inhibited autophosphorylation of aurora-A and -B as well as phosphorylation of histone H3 on Ser(10), a proximal substrate of aurora-B. The predominant cellular response of tumor cells to AMG 900 treatment was aborted cell division without a prolonged mitotic arrest, which ultimately resulted in cell death. AMG 900 inhibited the proliferation of 26 tumor cell lines, including cell lines resistant to the antimitotic drug paclitaxel and to other aurora kinase inhibitors (AZD1152, MK-0457, and PHA-739358), at low nanomolar concentrations. Furthermore, AMG 900 was active in an AZD1152-resistant HCT116 variant cell line that harbors an aurora-B mutation (W221L). Oral administration of AMG 900 blocked the phosphorylation of histone H3 in a dose-dependent manner and significantly inhibited the growth of HCT116 tumor xenografts. Importantly, AMG 900 was broadly active in multiple xenograft models, including 3 multidrug-resistant xenograft models, representing 5 tumor types. AMG 900 has entered clinical evaluation in adult patients with advanced cancers and has the potential to treat tumors refractory to anticancer drugs such as the taxanes.

Discovery of a potent, selective, and orally bioavailable pyridinyl-pyrimidine phthalazine aurora kinase inhibitor.

The discovery of aurora kinases as essential regulators of cell division has led to intense interest in identifying small molecule aurora kinase inhibitors for the potential treatment of cancer. A high-throughput screening effort identified pyridinyl-pyrimidine 6a as a moderately potent dual inhibitor of aurora kinases -A and -B. Optimization of this hit resulted in an anthranilamide lead (6j) that possessed improved enzyme and cellular activity and exhibited a high level of kinase selectivity. However, this anthranilamide and subsequent analogues suffered from a lack of oral bioavailability. Converting the internally hydrogen-bonded six-membered pseudo-ring of the anthranilamide to a phthalazine (8a-b) led to a dramatic improvement in oral bioavailability (38-61%F) while maintaining the potency and selectivity characteristics of the anthranilamide series. In a COLO 205 tumor pharmacodynamic assay measuring phosphorylation of the aurora-B substrate histone H3 at serine 10 (p-histone H3), oral administration of 8b at 50 mg/kg demonstrated significant reduction in tumor p-histone H3 for at least 6 h.

Adherent cell assay results affected by variable z-position mixing.

We demonstrate that modifying mixing dynamics after addition of organic solute into aqueous buffers dramatically affects cell morphology and protein expression. Variable z-position (VZP) or varying the height of aspiration and dispense positions during mixing eliminates artifactual effects. Here, we tested 4 adherent cell types and show effects of VZP on quantitative imaging, protein expression, viability, and morphology. The result: The quantitation of cytoplasmic fluorescence within the fields of interest of the phalloidin-actin stain assay improved by 47% and fluorescence variability emitted by cells expressing green fluorescence protein (GFP) fusion proteins decreased by 15%. Assays that perform measurement by averaged reading of the entire well are somewhat susceptible. For example, protein production decreased 8% on the hypoxia response element (HRE)-luciferase assay. VZP did not affect quantitative cell viability, deviate the half maximal effective dose concentration (EC(50)) values or alter expected curve patterns. VZP is a valuable systematic process for cellular assay workflows as it efficiently folds organic solute into the aqueous solution.

The proactive approach--is it worthwhile? A prospective controlled ergonomic intervention study in office workers.

OBJECTIVE: Does proactive ergonomics program enhance office worker health and productivity? METHOD: The investigation was conducted in connection with the move of 1500 office staff to a building with improved ergonomics. It was focused on associations between workstation features, working postures, musculoskeletal pain symptoms, and eye strain before and 18 months after implementation of a proactive ergonomic program. The outcomes were compared between the intervention and a similar reference group. RESULTS: Associations between improvement of postures and less musculoskeletal pain and eye strain were confirmed. A cross association between several features and postures and improved symptoms was noted, along with improved productivity. CONCLUSION: The study suggests that a proactive program adhering to the OSHA recommendations needs to include an individual workstation assessment to be effective in reducing symptoms and increasing productivity.

Amiloride-sensitive Na+ channels contribute to regulatory volume increases in human glioma cells.

Despite intensive research, brain tumors remain among the most difficult type of malignancies to treat, due largely to their diffusely invasive nature and the associated difficulty of adequate surgical resection. To migrate through the brain parenchyma and to proliferate, glioma cells must be capable of significant changes in shape and volume. We have previously reported that glioma cells express an amiloride- and psalmotoxin-sensitive cation conductance that is not found in normal human astrocytes. In the present study, we investigated the potential role of this ion channel to mediate regulatory volume increase in glioma cells. We found that the ability of the cells to volume regulate subsequent to cell shrinkage by hyperosmolar solutions was abolished by both amiloride and psalmotoxin 1. This toxin is thought to be a specific peptide inhibitor of acid-sensing ion channel (ASIC1), a member of the Deg/ENaC superfamily of cation channels. We have previously shown this toxin to be an effective blocker of the glioma cation conductance. Our data suggest that one potential role for this conductance may be to restore cell volume during the cell's progression thorough the cell cycle and while the tumor cell migrates within the interstices of the brain.

High-content screening analysis of the p38 pathway: profiling of structurally related p38alpha kinase inhibitors using cell-based assays.

The complexity of the p38 mitogen-activated protein kinase (MAPK) signaling pathway presents challenges to understanding the efficacy of p38 inhibitors. Biochemical recombinant kinase assays and tumor necrosis factor alpha (TNFalpha) secretion assays are typically used to evaluate p38alpha inhibitors, but they do not provide insight into proximal intracellular events. Stimulation of the pathway evokes a cascade of phosphorylation events, accompanied by movement of molecules to different cellular compartments. Herein, we describe the profiling and potency comparison of a large set of p38alpha inhibitors with a pyrimidinone, imidazopyrimidine, or triazolopyrimidine core against biochemical recombinant p38alpha kinase activity, lipopolysaccharide (LPS)-mediated TNFalpha secretion by THP-1 cells, and a set of cellular imaging assays in SW1353 chondrocytes and baby hamster kidney cells. These pathway assays included p38 phosphorylation, MAPK-activated protein kinase 2 translocation, and heat shock protein (HSP) 27 phosphorylation. We established that HSP27 phosphorylation correlates well with LPS-induced TNFalpha secretion, validating our cellular imaging assays. We also found that the choice of cells and inducer can profoundly affect cellular potency results. High-content analysis may reveal signaling details, enriching our understanding of the mechanism of action of p38alpha inhibitors.

Hepatitis A vaccine uptake in San Diego County: Hispanic children are better immunized.

BACKGROUND: Few studies have examined compliance with hepatitis A vaccination recommendations or factors likely to predict vaccination against hepatitis A virus. OBJECTIVES: To investigate hepatitis A coverage among 3- to 17.9-year-olds in San Diego County and examine predictors of child and adolescent hepatitis A immunization. DESIGN, SETTING, AND PARTICIPANTS: A total of 1455 participants completed a random-digit dial telephone survey that assessed hepatitis A immunization status of 3- to 17.9-year-old children from May 1 to June 24, 2003. Analysis was limited to the 983 respondents with available immunization records or verified immunization histories. MAIN OUTCOME MEASURES: Receipt of at least 1 hepatitis A vaccine administered on or after the child's second birthday and differences in the frequencies of vaccination based on vaccine availability, sex, ethnicity, type of health care provider, mother's highest level of education, and parental knowledge of the hepatitis A vaccine recommendation. RESULTS: Participant response rate was 77.1%. Among all respondents aged 3 to 17.9 years, 59% received at least 1 hepatitis A vaccine and 41% completed the 2-shot regimen. The adjusted odds that a child received at least 1 hepatitis A vaccine was 3.6 times greater among Hispanic children compared with non-Hispanic children. Other predictors of hepatitis A immunization included child's age, having a public health care provider, lower maternal education, and parental knowledge of the vaccine recommendation. CONCLUSIONS: Results challenge historical patterns of underimmunization among Hispanic children compared with white children. Public health education and community awareness should be sustained in Hispanic communities, but interventions are needed in non-Hispanic communities.

(4-Piperidinylphenyl)aminoethyl amides as a novel class of non-covalent cathepsin K inhibitors.

A series of (4-piperidinylphenyl)aminoethyl amides based on dipeptide anilines were synthesized and tested against cathepsin K, cathepsin L and cathepsin B. These new non-covalent inhibitors exhibited single-digit nM inhibition of the cysteine proteases. Compounds 3 and 7 demonstrated potency in both mouse and human osteoclast resorption assays.

Implementing a seventh grade vaccination law: school factors associated with completion of required immunizations.

OBJECTIVES: We investigated school factors associated with successful implementation of a seventh grade vaccination requirement. METHODS: The proportion of students vaccinated with hepatitis B vaccine and measles containing vaccine was determined from records of schools in San Diego County, California. A school survey identified compliance strategies. Analysis identified factors associated with coverage. RESULTS: In October 1999, 67.2% of 38,875 students had received the required vaccine doses. Of 315 schools, coverage was less than 40% in 60 schools and exceeded 80% in 111 schools. Factors associated with high coverage included private schools, early and frequent notice to parents, and, for public schools, higher overall socioeconomic status of students. CONCLUSIONS: In preparation for a middle school vaccination requirement, early and frequent notification of parents improves coverage. Schools with a high percentage of low socioeconomic status students may require extra resources to support implementation.