Final results of a single institution experience with a pediatric-based regimen, the augmented Berlin-Frankfurt-Münster, in adolescents and young adults with acute lymphoblastic leukemia, and comparison to the hyper-CVAD regimen.

Several studies reported improved outcomes of adolescents and young adults (AYA) with acute lymphoblastic leukemia (ALL) treated with pediatric-based ALL regimens. This prompted the prospective investigation of a pediatric Augmented Berlin-Frankfurt-Münster (ABFM) regimen, and its comparison with hyper-fractionated cyclophosphamide, vincristine, Adriamycin, and dexamethasone (hyper-CVAD) in AYA patients. One hundred and six AYA patients (median age 22 years) with Philadelphia chromosome- (Ph) negative ALL received ABFM from October 2006 through March 2014. Their outcome was compared to 102 AYA patients (median age 27 years), treated with hyper-CVAD at our institution. The complete remission (CR) rate was 93% with ABFM and 98% with hyper-CVAD. The 5-year complete remission duration (CRD) were 53 and 55%, respectively (P = 0.98). The 5-year overall survival (OS) rates were 60 and 60%, respectively. The MRD status on Day 29 and Day 84 of therapy was predictive of long-term outcomes on both ABFM and hyper-CVAD. Severe regimen toxicities with ABFM included hepatotoxicity in 41%, pancreatitis in 11%, osteonecrosis in 9%, and thrombosis in 19%. Myelosuppression-associated complications were most significant with hyper-CVAD. In summary, ABFM and hyper-CVAD resulted in similar efficacy outcomes, but were associated with different toxicity profiles, asparaginase-related with ABFM and myelosuppression-related with hyper-CVAD. Am. J. Hematol. 91:819-823, 2016. © 2016 Wiley Periodicals, Inc.

Augmented Berlin-Frankfurt-Münster therapy in adolescents and young adults (AYAs) with acute lymphoblastic leukemia (ALL).

BACKGROUND: Various trials have reported improved outcomes for adolescents and young adults (AYAs) with acute lymphoblastic leukemia (ALL) who received treatment with pediatric-based regimens. Those reports prompted the current investigation of the pediatric augmented Berlin-Frankfurt-Münster (ABFM) regimen in AYA patients. The results were compared with those from a similar population that received the hyperfractionated cyclophosphamide, vincristine, doxorubicin, and dexamethasone (hyper-CVAD) regimen. METHODS: Eighty-five patients ages 12 to 40 years who had Philadelphia chromosome (Ph)-negative ALL received the ABFM regimen from October 2006 through April 2012. Their outcome was compared with outcomes in 71 historic AYA patients who received hyper-CVAD from the authors' institution. Patient and disease characteristics, as well as minimal residual disease status, were analyzed for their impact on outcomes. RESULTS: The complete response rate with ABFM was 94%. The 3-year complete remission duration (CRD) and overall survival (OS) rates were 70% and 74%, respectively. For patients aged ≤21 years, the 3-year CRD and OS rates were 72% and 85%, respectively; and, for patients ages 21 to 40 years, the respective rates were 69% and 60%. The initial white blood cell count was an independent predictive factor of OS and CRD. The minimal residual disease status on days 29 and 84 of therapy also were predictive of long-term outcomes. Severe regimen toxicities included transient hepatotoxicity in 35% to 39% of patients, pancreatitis in 11% of patients, osteonecrosis in 11% of patients, and thrombosis in 22% of patients. The 3-year OS rate was 74% in the ABFM group versus 71% in the hyper-CVAD group, and the corresponding 3-year CRD rate was 70% versus 66%, respectively. CONCLUSIONS: ABFM was tolerable in AYA patients with ALL but was not associated with significant improvements in CRD and OS compared with hyper-CVAD.

Resurrection of a clinical antibody: template proteogenomic de novo proteomic sequencing and reverse engineering of an anti-lymphotoxin-α antibody.

A mouse hybridoma antibody directed against a member of the tumour necrosis factor (TNF)-superfamily, lymphotoxin-alpha (LT-α), was isolated from stored mouse ascites and purified to homogeneity. After more than a decade of storage the genetic material was not available for cloning; however, biochemical assays with the ascites showed this antibody against LT-α (LT-3F12) to be a preclinical candidate for the treatment of several inflammatory pathologies. We have successfully rescued the LT-3F12 antibody by performing MS analysis, primary amino acid sequence determination by template proteogenomics, and synthesis of the corresponding recombinant DNA by reverse engineering. The resurrected antibody was expressed, purified and shown to demonstrate the desired specificity and binding properties in a panel of immuno-biochemical tests. The work described herein demonstrates the powerful combination of high-throughput informatic proteomic de novo sequencing with reverse engineering to reestablish monoclonal antibody-expressing cells from archived protein sample, exemplifying the development of novel therapeutics from cryptic protein sources.

A phase 2 study of brentuximab vedotin in patients with CD30-positive advanced systemic mastocytosis.

There is an unmet need for effective therapies for advanced systemic mastocytosis (advSM). CD30 is expressed on the surface of neoplastic mast cells (MC) in more than 50% of patients with advSM. Brentuximab vedotin (BV) is a CD30-directed antibody-drug conjugate with preclinical evidence supporting both an antineoplastic effect and an attenuation of immunoglobulin E-associated mediator release. These observations are the basis for this phase 2 trial of BV monotherapy (1.8 mg/kg IV every 3 weeks up to 8 cycles) in patients with CD30-positive advSM. The primary objective was to determine the efficacy of BV according to International Working Group-Myeloproliferative Neoplasms Research and Treatment-European Competence Network on Mastocytosis (IWG-MRT-ECNM) response criteria. Secondary objectives included evaluation of safety, changes in bone marrow (BM) MC burden, serum tryptase level, flow cytometric quantification of MC surface expression of CD30, and self-reported symptom burden. The trial enrolled 10 patients with a diagnosis of CD30+ advSM (aggressive SM, SM with an associated hematologic neoplasm [SM-AHN], or mast cell leukemia [MCL]) with 1 or more signs of SM-related organ damage. According to IWG-MRT-ECNM criteria, none of the patients demonstrated better than stable disease with BV. In addition, there were no significant reductions in BM MC burden, serum tryptase levels, or MC surface expression of CD30. Self-reported symptom scores showed no durable improvement with BV treatment. We conclude that BV is not active as a single agent in CD30+ advSM. This trial was registered at www.clinicaltrials.gov as #NCT01807598.

A phase 2 study of pracinostat combined with ruxolitinib in patients with myelofibrosis.

Although ruxolitinib improves symptoms and splenomegaly in patients with advanced myelofibrosis, whether this agent is truly disease-modifying remains unclear. Histone deacetylase inhibitors (HDACi) downregulate JAK2 via interference with chaperone function. Pracinostat, a pan-HDACi, has modest single-agent activity in myelofibrosis. We conducted a single-institution, phase 2, investigator-initiated trial of ruxolitinib plus pracinostat (begun after 12 weeks of ruxolitinib) in 25 patients with myelofibrosis, of whom 20 received both agents. Sixteen (80%) patients had objective responses (all 'clinical improvement'). The rate of spleen response (by palpation) was 74%, and that of symptom response 80%. Most responses occurred prior to pracinostat initiation. Three patients experienced improvement in bone marrow fibrosis, and one a near-complete molecular response after two years on study treatment. All patients discontinued pracinostat and are currently off-study. Pracinostat interruptions and dose reductions were frequent, often due to worsening anemia. These findings do not support continued development of pracinostat in myelofibrosis.

Semiautomated Small-Scale Purification Method for High-Throughput Expression Analysis of Recombinant Proteins.

The expression analysis of recombinant proteins is a challenging step in any high-throughput protein production pipeline. Often multiple expression systems and a variety of expression construct designs are considered for the production of a protein of interest. There is a strong need to triage constructs rapidly and systematically. This chapter describes a semiautomated method for the simultaneous purification and characterization of proteins expressed from multiple samples of expression cultures from the E. coli, baculovirus expression vector system, and mammalian transient expression systems. This method assists in the selection of the most promising expression construct(s) or the most favorable expression condition(s) to move forward into large-scale protein production.

Phase II study of bi-weekly temozolomide plus bevacizumab for adult patients with recurrent glioblastoma.

PURPOSE: Bevacizumab is an active anti-angiogenic agent in the treatment of recurrent glioblastoma. Temozolomide can prolong survival in patients with newly diagnosed glioblastoma. At recurrence, alternate dosing of temozolomide has shown to further deplete methyl-guanine-methyltransferase (MGMT) conferring added activity for patients who have progressed on the standard dosing regimen. In this study, bevacizumab plus biweekly temozolomide was evaluated for efficacy in adult patients with recurrent glioblastoma. METHODS: Thirty patients with recurrent glioblastoma were treated with bevacizumab on (10 mg/kg i.v.) days 1 and 15 of a 28-day cycle combined with temozolomide (100 mg/m2) on days 1-5 and 15-19 on a 28-day cycle. Responses were assessed at week 4 and then every 8 weeks. MGMT status and quality of life measures were also assessed. RESULTS: Overall response rate from diagnosis was 51 weeks, the 6-month progression-free survival was 52%, and median time to tumor progression was 5.5 months. CONCLUSION: Bevacizumab plus bi-weekly temozolomide was well tolerated and may be a salvage regimen to be considered in a subset of patients with recurrent glioblastoma.

Vancomycin use in a rural hospital: a 3-year retrospective study.

INTRODUCTION: Urban centres often perform audits of vancomycin use as they face outbreaks of resistant organisms. We undertook this study to understand the indications and duration of intravenous vancomycin in a rural setting. METHODS: We conducted a retrospective chart audit for all patients who received intravenous vancomycin over a 3-year period at a rural hospital in northwestern Ontario. RESULTS: Vancomycin was used intravenously in 180 patients during the study period. It was used for short courses (median 3 d), and serum levels were below target 72% of the time. CONCLUSION: High rates of invasive methicillin-resistant Staphylococcus aureus bacteremia and limited antibiotic choices in the field likely contributed to short courses of this antibiotic. Further study on clinical severity and antibiotic choice is needed. Additionally, weight-based dosing may result in target serum levels being achieved more frequently.

INTRODUCTION: Les centres urbains effectuent souvent des vérifications de l'utilisation de la vancomycine en raison du risque d'éclosions d'infections causées par des agents pathogènes résistants. Nous avons entrepris cette étude pour comprendre les indications et la durée de l'antibiothérapie par vancomycine intraveineuse en région rurale. MÉTHODES: Nous avons procédé à une analyse rétrospective des dossiers de tous les patients qui ont reçu de la vancomycine intraveineuse sur une période de 3 ans dans un hôpital rural du Nord-Ouest de l'Ontario. RÉSULTATS: La vancomycine a été administrée par voie intraveineuse chez 180 patients durant la période de l'étude. Elle a été utilisée pendant de courtes périodes (durée médiane 3 jours) et les taux sériques étaient inférieurs aux taux ciblés 72 % du temps. CONCLUSION: Les taux élevés de bactériémie invasive à Staphylococcus aureus méthicillinorésistant et le choix limité d'antibiotiques sur le terrain ont probablement contribué à la brièveté des antibiothérapies avec cet agent. Des études plus approfondies sur la gravité des cas cliniques et le choix des antibiotiques s'imposent. De plus, l'établissement de la dose en fonction du poids corporel pourrait favoriser l'atteinte plus fréquente des taux sériques cibles.

Identifying student misconceptions in biomedical course assessments in dental education.

Dental student performance on examinations has traditionally been estimated by calculating the percentage of correct responses rather than by identifying student misconceptions. Although misconceptions can impede student learning and are refractory to change, they are seldom measured in biomedical courses in dental schools. Our purpose was to determine if scaling student confidence and the clinical impact of incorrect answers could be used on multiple-choice questions (MCQs) to identify potential student misconceptions. To provide a measure of student misconception, faculty members indicated the correct answer on twenty clinically relevant MCQs and noted whether the three distracters represented potentially benign, inappropriate, or harmful application of student knowledge to patient treatment. A group of 105 third-year dental students selected what they believed was the most appropriate answer and their level of sureness (1 to 4 representing very unsure, unsure, sure, and very sure) about their answer. Misconceptions were defined as sure or very sure incorrect responses that could result in inappropriate or harmful clinical treatment. In the results, 5.2 percent of the answers represented student misconceptions, and 74 percent of the misconceptions were from four case-based interpretation questions. The mean student sureness was 3.6 on a 4.0 scale. The students' sureness was higher with correct than with incorrect answers (p<0.001), yet there was no difference in sureness levels among their incorrect (benign, inappropriate, or harmful) responses (p>0.05). This study found that scaling student confidence and clinical impact of incorrect answers provided helpful insights into student thinking in multiple-choice assessment.

Targeting FGFR4 inhibits hepatocellular carcinoma in preclinical mouse models.

The fibroblast growth factor (FGF)-FGF receptor (FGFR) signaling system plays critical roles in a variety of normal developmental and physiological processes. It is also well documented that dysregulation of FGF-FGFR signaling may have important roles in tumor development and progression. The FGFR4-FGF19 signaling axis has been implicated in the development of hepatocellular carcinomas (HCCs) in mice, and potentially in humans. In this study, we demonstrate that FGFR4 is required for hepatocarcinogenesis; the progeny of FGF19 transgenic mice, which have previously been shown to develop HCCs, bred with FGFR4 knockout mice fail to develop liver tumors. To further test the importance of FGFR4 in HCC, we developed a blocking anti-FGFR4 monoclonal antibody (LD1). LD1 inhibited: 1) FGF1 and FGF19 binding to FGFR4, 2) FGFR4-mediated signaling, colony formation, and proliferation in vitro, and 3) tumor growth in a preclinical model of liver cancer in vivo. Finally, we show that FGFR4 expression is elevated in several types of cancer, including liver cancer, as compared to normal tissues. These findings suggest a modulatory role for FGFR4 in the development and progression of hepatocellular carcinoma and that FGFR4 may be an important and novel therapeutic target in treating this disease.

Understanding sociocultural and psychological factors affecting transgender people of color in San Francisco.

This ethnographic qualitative study explored the needs of transgender people of color, including biological transitioning issues, gender and group membership identity formation, HIV, and other health issues. The sample consisted of transgender youth and adults of color in San Francisco (N = 43). Data were collected from in-depth interviews with 20 youth and adults and focus groups with 23 individuals. The study focused on perspectives of racial and ethnic minorities from Asian/Pacific Islander, African American, and Latino backgrounds. The medical decision-making perspective was used to gain a deeper understanding of sociocultural and psychological factors affecting transgender individuals of color in San Francisco. The major themes that emerged were gender identity, group membership, transitioning and related issues, sex work, alcohol and drug use, mental health and health care, sense of community, HIV, resources, and other support. Key clinical considerations that health providers can use to improve care of transgender individuals of color are included.

Acción Mutua (Shared Action): a multipronged approach to delivering capacity-building assistance to agencies serving Latino communities in the United States.

Culturally appropriate, theory-based capacity-building assistance can serve a vital role in helping HIV prevention providers remain up-to-date, effective, and responsive to those they serve. Funded by the Centers for Disease Control and Prevention (CDC), AIDS Project Los Angeles, in collaboration with San Francisco State University's César E. Chávez Institute, conducted full-day site visits and qualitative interviews in 2005 with mid-level management staff of CDC-funded community-based organizations delivering HIV prevention services to Latino communities in the western region of the United States. We found that agencies we visited (1) had not yet adapted the evidence-based interventions they were using at the time of our visit and (2) requested technical assistance and training in the areas of program development, evaluation, group facilitation techniques, consumer recruitment, client retention, intervention adaptation, and materials development. Findings from this needs assessment were used to inform our seven-pronged approach to delivering capacity-building assistance entitled "Acción Mutua" (Shared Action). The approach emphasizes strategic partnerships, stakeholder involvement, organizational self-assessment, culturally appropriate materials development, interactive training, tailored onsite technical assistance, and professional networking opportunities. This article describes our approach in detail, the assessment process we used to develop it, and its implications for capacity-building practice.