A poised fragment library enables rapid synthetic expansion yielding the first reported inhibitors of PHIP(2), an atypical bromodomain

Research into the chemical biology of bromodomains has been driven by the development of acetyl-lysine mimetics. The ligands are typically anchored by binding to a highly conserved asparagine residue. Atypical bromodomains, for which the asparagine is mutated, have thus far proven elusive targets, including PHIP(2) whose parent protein, PHIP, has been linked to disease progression in diabetes and cancers. The PHIP(2) binding site contains a threonine in place of asparagine, and solution screening have yielded no convincing hits. We have overcome this hurdle by combining the sensitivity of X-ray crystallography, used as the primary fragment screen, with a strategy for rapid follow-up synthesis using a chemically-poised fragment library, which allows hits to be readily modified by parallel chemistry both peripherally and in the core. Our approach yielded the first reported hit compounds of PHIP(2) with measurable IC50 values by an AlphaScreen competition assay. The follow-up libraries of four poised fragment hits improved potency into the sub-mM range while showing good ligand efficiency and detailed structural data

Persistence to Anti-CGRP Monoclonal Antibodies and onabotulinumtoxinA Among Patients With Migraine: A Retrospective Cohort Study

BACKGROUND: To date, real-world evidence on persistence to anti-calcitonin gene-related peptide (anti-CGRP) monoclonal antibodies (mAbs) or onabotulinumtoxinA have excluded eptinezumab. This retrospective cohort study was performed to compare treatment persistency among patients with migraine on anti-CGRP mAbs (erenumab, fremanezumab, galcanezumab, or eptinezumab) or onabotulinumtoxinA. METHODS: This retrospective study used IQVIA PharmMetrics data. Adult patients with migraine treated with an anti-CGRP mAb or onabotulinumtoxinA who had 12 months of continuous insurance enrollment before starting treatment were included. A most recent treatment episode analysis was used in which the most recent episode was defined as the latest treatment period with the same drug (anti-CGRP mAb or onabotulinumtoxinA) without a ≥ 15-day gap in medication supply on/after June 25, 2020, to December 31, 2021. Patients were indexed at the start of their most recent episode. Patients were considered non-persistent and discontinued the therapy associated with their most recent episode if there was ≥ 15-day gap in medication supply. A Cox proportional-hazards model estimated the discontinuation hazard between treatments. The gap periods and cohort definition were varied in sensitivity analyses. RESULTS: The study included 66,576 patients (median age 46 years, 88.6% female). More eptinezumab-treated patients had chronic migraine (727/1074), ≥ 3 previous acute (323/1074) or preventive (333/1074) therapies, and more prior treatment episodes (3) than other treatment groups. Based on a 15-day treatment gap, patients on subcutaneous anti-CGRP mAbs had a 32% (95% CI: 1.19, 1.49; erenumab), 42% (95% CI: 1.27, 1.61; galcanezumab), and 58% (95% CI: 1.42, 1.80; fremanezumab) higher discontinuation hazard than those receiving eptinezumab, with this relationship attenuated, but still statistically significant based on 30-day and 60-day treatment gaps. There was no significant difference in the discontinuation hazard between eptinezumab and onabotulinumtoxinA. Based on a 15-day treatment gap among patients who newly initiated therapy, the discontinuation hazard of subcutaneous anti-CGRP mAbs remained significantly higher compared to eptinezumab and onabotulinumtoxinA. CONCLUSION: Patients treated with eptinezumab demonstrated persistency that was higher than subcutaneous anti-CGRP mAbs and similar to onabotulinumtoxinA

Sequencing of diverse mandarin, pummelo and orange genomes reveals complex history of admixture during citrus domestication

Cultivated citrus are selections from, or hybrids of, wild progenitor species whose identities and contributions to citrus domestication remain controversial. Here we sequence and compare citrus genomes-a high-quality reference haploid clementine genome and mandarin, pummelo, sweet-orange and sour-orange genomes-and show that cultivated types derive from two progenitor species. Although cultivated pummelos represent selections from one progenitor species, Citrus maxima, cultivated mandarins are introgressions of C. maxima into the ancestral mandarin species Citrus reticulata. The most widely cultivated citrus, sweet orange, is the offspring of previously admixed individuals, but sour orange is an F1 hybrid of pure C. maxima and C. reticulata parents, thus implying that wild mandarins were part of the early breeding germplasm. A Chinese wild 'mandarin' diverges substantially from C. reticulata, thus suggesting the possibility of other unrecognized wild citrus species. Understanding citrus phylogeny through genome analysis clarifies taxonomic relationships and facilitates sequence-directed genetic improvement. (Résumé d'auteur

A Pandemic Influenza H1N1 Live Vaccine Based on Modified Vaccinia Ankara Is Highly Immunogenic and Protects Mice in Active and Passive Immunizations

The development of novel influenza vaccines inducing a broad immune response is an important objective. The aim of this study was to evaluate live vaccines which induce both strong humoral and cell-mediated immune responses against the novel human pandemic H1N1 influenza virus, and to show protection in a lethal animal challenge model.For this purpose, the hemagglutinin (HA) and neuraminidase (NA) genes of the influenza A/California/07/2009 (H1N1) strain (CA/07) were inserted into the replication-deficient modified vaccinia Ankara (MVA) virus - a safe poxviral live vector – resulting in MVA-H1-Ca and MVA-N1-Ca vectors. These live vaccines, together with an inactivated whole virus vaccine, were assessed in a lung infection model using immune competent Balb/c mice, and in a lethal challenge model using severe combined immunodeficient (SCID) mice after passive serum transfer from immunized mice. Balb/c mice vaccinated with the MVA-H1-Ca virus or the inactivated vaccine were fully protected from lung infection after challenge with the influenza H1N1 wild-type strain, while the neuraminidase virus MVA-N1-Ca induced only partial protection. The live vaccines were already protective after a single dose and induced substantial amounts of neutralizing antibodies and of interferon-γ-secreting (IFN-γ) CD4- and CD8 T-cells in lungs and spleens. In the lungs, a rapid increase of HA-specific CD4- and CD8 T cells was observed in vaccinated mice shortly after challenge with influenza swine flu virus, which probably contributes to the strong inhibition of pulmonary viral replication observed. In addition, passive transfer of antisera raised in MVA-H1-Ca vaccinated immune-competent mice protected SCID mice from lethal challenge with the CA/07 wild-type virus.The non-replicating MVA-based H1N1 live vaccines induce a broad protective immune response and are promising vaccine candidates for pandemic influenza

Sequencing of diverse mandarin, pummelo and orange genomes reveals complex history of admixture during citrus domestication

Cultivated citrus are selections from, or hybrids of, wild progenitor species whose identities and contributions to citrus domestication remain controversial. Here we sequence and compare citrus genomes-a high-quality reference haploid clementine genome and mandarin, pummelo, sweet-orange and sour-orange genomes-and show that cultivated types derive from two progenitor species. Although cultivated pummelos represent selections from one progenitor species, Citrus maxima, cultivated mandarins are introgressions of C. maxima into the ancestral mandarin species Citrus reticulata. The most widely cultivated citrus, sweet orange, is the offspring of previously admixed individuals, but sour orange is an F1 hybrid of pure C. maxima and C. reticulata parents, thus implying that wild mandarins were part of the early breeding germplasm. A Chinese wild 'mandarin' diverges substantially from C. reticulata, thus suggesting the possibility of other unrecognized wild citrus species. Understanding citrus phylogeny through genome analysis clarifies taxonomic relationships and facilitates sequence-directed genetic improvement

Disparities in Healthcare Expenditure and Mortality by Neighborhood Income in Chronic Myeloid Leukemia

INTRODUCTION: Tyrosine-kinase inhibitors (TKIs) for treatment of chronic myeloid leukemia (CML) is financially burdensome. Patients with a low neighborhood income, whose treatment cost is high relative to or exceeds income, may become “priced out” of treatment resulting in decreased TKI initiation and an increased rate of death. Low neighborhood income patients may also forgo non-cancer treatment to maintain cancer treatment. We evaluated the trend in TKI drug price and out-of-pocket (OOP) expenditure across time, and whether there is a disparity in mortality and non-cancer healthcare expenditure between patients with high and low neighborhood income. METHODS: We used the Surveillance, Epidemiology, and End Results (SEER) – Medicare database to conduct our analyses. We calculated annual average TKI gross-payment and monthly OOP expenditure and compared annual changes to inflation indices. Low neighborhood income was defined as a census-tract income below the median. We measured the adjusted effect of neighborhood income on the rate of TKI initiation using a Cox-proportional hazards model. Similarly, among TKI initiators, we measured the effect of neighborhood income on the rate of mortality. We used inverse probability weighted partitioned regression to measure the adjusted association between neighborhood income and cumulative non-cancer expenditure over 60-months. RESULTS: Average annual gross payment per 30-day supply of a TKI increased by an average of 12.8% annually throughout the observation, which exceeded 2% medical inflation. There was no trend in median annual OOP expenditure per 30-day supply, which varied between $450 to$600. We found that of 503 CML cases, low neighborhood income was not associated with TKI initiation (HR=1.12, P=0.48). Among 354 TKI initiators, low neighborhood income was associated with an increased rate of CML-specific death (HR=3.24, P=0.04), and patients with low neighborhood income had an insignificant cumulative total non-cancer expenditure that was $99,110 (95$659,485 to $275,113]) less. CONCLUSION: Escalating TKI price and high OOP expenditure places financial pressure on patients, which may manifest as an increased risk of CML-specific death in low-income patients, but not as forgoing non-cancer treatment. Patients may instead be forgoing unmeasured goods and services. Our findings support the need for policy interventions which mitigate the financial burden of cancer

Economic Evaluations of Clinical Pharmacy Services in the United States: 2011-2017

Studies evaluating the cost-effectiveness of clinical pharmacy services (CPS) are needed to justify implementation and reimbursement. Through a systematic review, we describe services provided by pharmacists and their economic outcomes. We conducted a literature search of published studies in PubMed, Ovid, and Embase from January 2011 through December 2017. Manuscripts evaluating a CPS with patient-level economic outcomes and conducted in the United States were included. Study risks of bias were classified by study design characteristics. Economic evaluations were classified according to the presence of a comparator, and cost and outcome measures included. The quality of full economic evaluations was assessed using the Quality of Health Economic Studies (QHES) instrument. Descriptive statistics were used to summarize CPS characteristics. After screening, 115 studies were included. Type of service provided included general pharmacotherapy (41%), disease management (30%), and targeted drug program (17%). Settings included hospital (34%), ambulatory care (28%), and community pharmacy (17%). Study designs were considered high risk of bias (use of a historical control group or no control group) in 69% of cases while 25% were medium risk of bias (non-randomized with a concurrent control group) and 6% were low risk of bias (randomized experimental or multigroup interrupted time series). Economic evaluation types were descriptive studies that measured cost and/or outcomes of a CPS (55%), comparative studies that measured cost or outcomes of a CPS and a comparator (37%), and full evaluations that measured cost and outcomes of a CPS and a comparator (8%). Among nine full evaluations, the median (range) QHES score was 74 (59-95) and four reported the CPS as being more effective at a lower cost. Few full economic evaluations were conducted, but supported the cost-effectiveness of CPS. Use of a comparator group and measurement of economic inputs and outcomes would strengthen the body of evidence

Defining totipotency using criteria of increasing stringency

Abstract Totipotency is the ability of a single cell to give rise to all the differentiated cells that build the conceptus, yet how to capture this property in vitro remains incompletely understood. Defining totipotency relies upon a variety of assays of variable stringency. Here we describe criteria to define totipotency. We illustrate how distinct criteria of increasing stringency can be used to judge totipotency by evaluating candidate totipotent cell types in the mouse, including early blastomeres and expanded or extended pluripotent stem cells. Our data challenge the notion that expanded or extended pluripotent states harbor increased totipotent potential relative to conventional embryonic stem cells under in vivo conditions.status: publishe