The Breastfeeding Self-Efficacy Scale-Short Form (BSES-SF): German Translation and Psychometric Assessment.

BACKGROUND: German-speaking mothers have breastfeeding rates below the international breastfeeding recommendations. Previous research has found that breastfeeding self-efficacy is an important and modifiable predictor of breastfeeding outcomes, thus improving breastfeeding rates. The Breastfeeding Self-Efficacy Scale-Short Form (BSES-SF) is used in many countries to assess maternal breastfeeding self-efficacy. This instrument has not been available in German. RESEARCH AIMS: To translate the BSES-SF into German and assess its psychometric properties among breastfeeding mothers up to 12 weeks postpartum. METHODS: This cross-sectional study was conducted online with 355 breastfeeding mothers recruited from breastfeeding groups through Facebook. The BSES-SF was translated into German using forward and back-translation. To test reliability, item-total characteristics, including Cronbach's alpha, were examined. We used principal component analysis, as well as known-groups comparisons for evaluating construct validity, and examined the relationship between breastfeeding self-efficacy and demographic variables. RESULTS: The mean age of participants was 32.4 years (SD = 4.32). The Cronbach's alpha coefficient was .88 and corrected item-total correlations ranged between .37 and .73. Principal components analysis yielded one component with factor loadings >.40 and an eigenvalue of 5.62, which explained 40% of the total variance. In addition, known group comparisons provided further evidence for construct validity. There was no significant difference in BSES-SF scores in terms of demographic and obstetrics characteristics. CONCLUSION: Our results provide evidence that the German version of the BSES-SF is a reliable and valid tool for measuring breastfeeding self-efficacy among mothers in German-speaking countries.

Short-term dynamics of loneliness and depressive symptoms: Gender differences in older adults.

BACKGROUND: Previous research examining the relationship between loneliness and depressive symptoms often treated these constructs as static traits rather than dynamic states. The current study focused on the short-term, prospective link between loneliness and depressive symptoms, while also analyzing potential gender differences. METHODS: We modeled panel data from seven bi-weekly assessments gathered in the FRequent health Assessment In Later life (FRAIL70+) study. At baseline, the sample size amounted to N = 426 community-dwelling older adults aged 70 years or older in Austria. The relationship between loneliness and depressive symptoms was analyzed using a latent change score modeling framework. RESULTS: As regards depressive symptoms, women showed higher initial levels and more change across the three months than men. Loneliness did not considerably change across time for both sexes. Moreover, greater levels of loneliness at a given point in time were associated with an accelerated increase in depressive symptoms two weeks later in women but not in men. CONCLUSION: Loneliness appeared to be a potential determinant of future increases in depressive symptoms. The varying effects observed between men and women suggest potential gender differences in short-term fluctuations of depressive symptoms and their underlying mechanisms.

Frailty predicts all-cause and cause-specific mortality among older adults in Austria: 8-year mortality follow-up of the Austrian Health Interview Survey (ATHIS 2014).

BACKGROUND: The frailty index (FI) is an established predictor of all-cause mortality among older adults, but less is known with regard to cause-specific mortality, and whether the predictive power of the FI varies between men and women and by socio-economic position. METHODS: We assessed all-cause and cause-specific mortality during 8 years of follow-up (median = 7 years) among the population-representative sample of older adults (65 + , n = 2,561) from the European Health Interview Survey in Austria (ATHIS 2014). A FI at baseline was constructed from 41 health deficits. Official cause of death information from Statistics Austria was linked with the survey data by the Austrian Micro Data Center (AMDC). Next to all-cause mortality, we differentiated between mortality from cardiovascular diseases (CVD), cancer, and other causes. Cox proportional hazard models adjusted for socio-demographic variables and causes of death as competing risks were used to assess mortality prediction. RESULTS: Among the participants, 43.5% were robust (FI < 0.10), 37.7% pre-frail (FI = 0.10-0.21), and 18.7% were frail (FI > 0.21). 405 (15.8%) participants died during follow-up. Among the deceased, 148 (36.5%) died from CVD, 127 (31.4%) died from cancer, and 130 (32.1%) died from other causes of death. The FI predicted all-cause (hazard ratio, HR = 1.33 per 0.1 FI and HR = 2.4 for frail compared to robust older adults) and cause-specific mortality risk (HRCVD = 1.25/2.46, HRcancer = 1.19/1.47, HRother = 1.49/3.59). Area under the curve (AUC) values were acceptable for CVD mortality (0.78) and other causes of death (0.74), and poor for cancer mortality (0.64). CONCLUSIONS: The FI predicts all-cause and cause-specific mortality (CVD, other causes) well, which points to its relevance as a potential screening tool for risk stratification among community-dwelling older adults.

Impact of Rehabilitation on Physical and Neuropsychological Health of Patients Who Acquired COVID-19 in the Workplace.

Workers, especially healthcare workers, are exposed to an increased risk for SARS-CoV-2 infection. However, less is known about the impact of rehabilitation on health outcomes associated with post-COVID. This longitudinal observational study examined the changes in physical and neuropsychological health and work ability after inpatient rehabilitation of 127 patients (97 females/30 males; age 21-69 years; Mean = 50.62) who acquired COVID-19 in the workplace. Post-COVID symptoms, functional status, physical performance, neuropsychological health, employment, and work ability were assessed before and after rehabilitation. Group differences relating to sex, professions, and acute COVID status were also analyzed. Except for fatigue, the prevalence of all post-COVID symptoms decreased after rehabilitation. Significant improvements in physical performance and neuropsychological health outcomes were determined. Moreover, healthcare workers showed a significantly greater reduction in depressive symptoms compared to non-healthcare workers. Nevertheless, participants reported poor work ability, and 72.5% of them were still unable to work after discharge from rehabilitation. As most participants were still suffering from the impact of COVID-19 at rehabilitation discharge, ongoing strategies in aftercare are necessary to improve their work ability. Further investigations of this study population at 6 and 12 months after rehabilitation should examine the further course of post-COVID regarding health and work ability status.

Comprehensive molecular characterization of a rare case of Philadelphia chromosome-positive acute myeloid leukemia.

The Philadelphia chromosome (Ph) resulting from the t(9;22) translocation generates the oncogenic BCR::ABL1 fusion protein that is most commonly associated with chronic myeloid leukemia (CML) and Ph-positive (Ph+) acute lymphoblastic leukemia (ALL). There are also rare instances of patients (≤1%) with newly diagnosed acute myeloid leukemia (AML) that harbor this translocation (Paietta et al., Leukemia 12: 1881 [1998]; Keung et al., Leuk Res 28: 579 [2004]; Soupir et al., Am J Clin Pathol 127: 642 [2007]). AML with BCR::ABL has only recently been provisionally classified by the World Health Organization as a diagnostically distinct subtype of AML. Discernment from the extremely close differential diagnosis of myeloid blast crisis CML is challenging, largely relying on medical history rather than clinical characteristics (Arber et al., Blood 127: 2391 [2016]). To gain insight into the genomic features underlying the evolution of AML with BCR::ABL, we identified a patient presenting with a high-risk myelodysplastic syndrome that acquired a BCR::ABL alteration after a peripheral blood stem cell transplant. Serial samples were collected and analyzed using whole-exome sequencing, RNA-seq, and ex vivo functional drug screens. Persistent subclones were identified, both at diagnosis and at relapse, including an SF3B1p.Lys700Glu mutation that later cooccurred with an NRASp.Gly12Cys mutation. Functional ex vivo drug screening performed on primary patient cells suggested that combination therapies of ABL1 with RAS or PI3K pathway inhibitors could have augmented the patient's response throughout the course of disease. Together, our findings argue for the importance of genomic profiling and the potential value of ABL1 inhibitor-inclusive combination treatment strategies in patients with this rare disease.

Integrative analysis of drug response and clinical outcome in acute myeloid leukemia.

Acute myeloid leukemia (AML) is a cancer of myeloid-lineage cells with limited therapeutic options. We previously combined ex vivo drug sensitivity with genomic, transcriptomic, and clinical annotations for a large cohort of AML patients, which facilitated discovery of functional genomic correlates. Here, we present a dataset that has been harmonized with our initial report to yield a cumulative cohort of 805 patients (942 specimens). We show strong cross-cohort concordance and identify features of drug response. Further, deconvoluting transcriptomic data shows that drug sensitivity is governed broadly by AML cell differentiation state, sometimes conditionally affecting other correlates of response. Finally, modeling of clinical outcome reveals a single gene, PEAR1, to be among the strongest predictors of patient survival, especially for young patients. Collectively, this report expands a large functional genomic resource, offers avenues for mechanistic exploration and drug development, and reveals tools for predicting outcome in AML.

Strengths and weaknesses of the German translation of the Inflexible Eating Questionnaire and of eating disorder assessment in general.

Objective: The present article introduces the German translation of the Inflexible Eating Questionnaire (IEQ-G), performs a psychometric evaluation, and explores the relationship of Inflexible Eating to the subscales of the Eating Disorder Examination-Questionnaire (EDE-Q) and Obsessive-Compulsive (OC) symptoms. Methods: The cross-sectional study was carried out in the German-speaking area. A paper and pencil survey was completed by 612 females and 442 males of the general population. Results: SEM analyses showed that the IEQ-G allows for calculating a total score and invariance tests were mostly promising. As a side result, the original 4-factorial structure of the EDE-Q could not be replicated, but a 3 dimensional solution proved convincing. From a psychometric point of view, the IEQ-G outperformed the EDE-Q. On a latent level, Inflexible Eating was remarkably strong related to OC-symptoms and the EDE-Q subscales. Discussion: The detail analyses revealed that Eating Disorder assessment in general lacks subgroup-specific aspects, for instance, regarding gender or dietary preferences, important for early diagnosis and screening of ED. The IEQ-G proved applicable in a German speaking adult population and recommends itself for cross-cultural studies.

Cancer-Associated Fibroblasts Modulate Transcriptional Signatures Involved in Proliferation, Differentiation and Metastasis in Head and Neck Squamous Cell Carcinoma.

Cancer-associated fibroblasts (CAFs) are known to increase tumor growth and to stimulate invasion and metastasis. Increasing evidence suggests that CAFs mediate response to various treatments. HNSCC cell lines were co-cultured with their patient-matched CAFs in 2D and 3D in vitro models, and the tumor cell gene expression profiles were investigated by cDNA microarray and qRT-PCR. The mRNA expression of eight candidate genes was examined in tumor biopsies from 32 HNSCC patients and in five biopsies from normal oral tissue. Differences in overall survival (OS) were tested with Kaplan-Meier long-rank analysis. Thirteen protein coding genes were found to be differentially expressed in tumor cells co-cultured with CAFs in 2D and 81 in 3D when compared to tumor cells cultured without CAFs. Six of these genes were upregulated both in 2D and 3D (POSTN, GREM1, BGN, COL1A2, COL6A3, and COL1A1). Moreover, two genes upregulated in 3D, MMP9 and FMOD, were significantly associated with the OS. In conclusion, we demonstrated in vitro that CAF-derived signals alter the tumor cell expression of multiple genes, several of which are associated with differentiation, epithelial-to-mesenchymal transition (EMT) phenotype, and metastasis. Moreover, six of the most highly upregulated genes were found to be overexpressed in tumor tissue compared to normal tissue.

DNA methylation epitypes highlight underlying developmental and disease pathways in acute myeloid leukemia.

Acute myeloid leukemia (AML) is a molecularly complex disease characterized by heterogeneous tumor genetic profiles and involving numerous pathogenic mechanisms and pathways. Integration of molecular data types across multiple patient cohorts may advance current genetic approaches for improved subclassification and understanding of the biology of the disease. Here, we analyzed genome-wide DNA methylation in 649 AML patients using Illumina arrays and identified a configuration of 13 subtypes (termed "epitypes") using unbiased clustering. Integration of genetic data revealed that most epitypes were associated with a certain recurrent mutation (or combination) in a majority of patients, yet other epitypes were largely independent. Epitypes showed developmental blockage at discrete stages of myeloid differentiation, revealing epitypes that retain arrested hematopoietic stem-cell-like phenotypes. Detailed analyses of DNA methylation patterns identified unique patterns of aberrant hyper- and hypomethylation among epitypes, with variable involvement of transcription factors influencing promoter, enhancer, and repressed regions. Patients in epitypes with stem-cell-like methylation features showed inferior overall survival along with up-regulated stem cell gene expression signatures. We further identified a DNA methylation signature involving STAT motifs associated with FLT3-ITD mutations. Finally, DNA methylation signatures were stable at relapse for the large majority of patients, and rare epitype switching accompanied loss of the dominant epitype mutations and reversion to stem-cell-like methylation patterns. These results show that DNA methylation-based classification integrates important molecular features of AML to reveal the diverse pathogenic and biological aspects of the disease.

Rehabilitation and Return-to-Work of Patients Acquiring COVID-19 in the Workplace: A Study Protocol for an Observational Cohort Study.

Background: In 2020, the novel coronavirus disease (COVID-19) developed into a worldwide pandemic. The course of COVID-19 is diverse, non-specific, and variable: Affected persons suffer from physical, cognitive, and psychological acute and long-term consequences. The symptoms influence everyday life activities, as well as work ability in the short or long-term. Healthcare professionals are considered particularly vulnerable to COVID-19 compared to the general population. In Germany, COVID-19 is recognized as an occupational disease or a work-related accident under certain conditions. Disease-specific rehabilitation is recommended for patients following acute COVID-19 to recover physical and neuropsychological performance and to improve work ability. Currently, there are limited findings on the short-term or long-term impact of COVID-19 as a recognized occupational disease or work-related accident, as well as on rehabilitation programs and associated influencing factors. Thus, the present research project will investigate these questions. Methods: For this observational cohort study, post-acute patients with COVID-19 as a recognized occupational disease or work-related accident according to the insurance regulations for COVID-19 will be recruited at the BG Hospital for Occupational Disease in Bad Reichenhall, Germany. All participants will complete a comprehensive multimodal and interdisciplinary inpatient rehabilitation program for a duration of at least 3 weeks, beginning after their acute COVID-19 infection and depending on their individual indication and severity of disease. Participants will complete medical, functional, motor, psychological, and cognitive measurements at four time points (at the beginning (T1) and end (T2) of inpatient rehabilitation; 6 (T3) and 12 (T4) months after the beginning of inpatient rehabilitation). Discussion: The present research project will help to assess and describe long-term effects of COVID-19 as a recognized occupational disease or work-related accident on physical and neuropsychological health, as well as on everyday activities and work ability of affected insured persons. In addition, this study will investigate influencing factors on severity and course of COVID-19. Furthermore, we will examine rehabilitation needs, measures, occurring specifics, and the feasibility of the rehabilitation procedure and disease development in the patients. The results of the intended study will further advance common recommendations for targeted and tailored rehabilitation management and participation in inpatient rehabilitation. Clinical Trial Registration: www.drks.de, identifier: DRKS00022928.

Application of an Optimized Direct Lysis Method for Viral RNA Extraction Linking Contaminated Dates to Infection With Hepatitis A Virus.

Consumption of dates has not been considered a common risk of hepatitis A virus (HAV) infection. In January 2018, an outbreak of hepatitis was identified with cases resident in all regions of Denmark. All the detected strains belonged to HAV genotype 3A. Epidemiological investigations through patients' interviews, case-control and trace-back studies pointed toward different batches of dates from a single producer as the vehicle of infection. Boxes of dates from suspected batches were collected from homes of patients and healthy families and analyzed using a recently reported optimized direct lysis method, consisting of simultaneous viral RNA elution and extraction from dates followed by purification of the nucleic acids. Extracts were analyzed for HAV and norovirus (NoV) RNA using RT-qPCR, while detected HAV were genotyped by Sanger sequencing. Among 20 nucleic acid extracts representing eight batches of dates, RNA of HAV (9.3 × 102 genome copies/g) and NoV genogroup (G)II (trace amounts) were detected in one batch, while NoV GII RNA (trace amounts) was detected in another. Average extraction efficiency of spiked process control murine norovirus was 20 ± 13% and the inhibitions of RT-qPCR detection of NoV GI, NoV GII, and HAV were 31 ± 34, 9 ± 9, and 3 ± 7%, respectively. The HAV genome detected in the dates matched by sequence 100% to the HAV genotype 3A detected in stool samples from cases implicated in the outbreak. This confirmed, to our knowledge, for the first time a sequence link between HAV infection and consumption of contaminated dates, suggesting dates to be an important vehicle of HAV transmission.

Status of risk-based approach and national framework for safe drinking water in small water supplies of the Nordic water sector.

Reliable safe water supply is a pillar of society and a key to public health. The Nordic countries have an abundance of clean fresh water as a source for drinking water supplies. They have followed developments in safeguarding water, both the recommendations of the World Health Organization framework for safe drinking water and European legislation. Worldwide, including the Nordic countries, small water supplies are less compliant with water safety regulation. The forthcoming EU directive on drinking water require risk-based approaches and improved transparency on water quality. This research looks at the Nordic frameworks for safe water supply, with emphasis on risk-based approaches and smaller systems. We analyzed the legal frameworks for safe water, the structure of the water sector across the Nordic countries and explored how prepared these countries are to meet these requirements. Our findings show that, while legal requirements are mostly in place, delivery of information to the public needs to be improved. Most Nordic countries are in the process of implementing risk-based management in large and medium size water supplies, whereas small supplies are lagging. We conclude that a key to success is increased training and support for small supplies. We suggest wider adoption of the Nordic model of cooperation with benchmarking of safe water for all to transfer knowledge between the countries. This work provides insights into challenges and opportunities for the Nordic countries and provides insights relevant to countries worldwide in their effort towards realization of SDG Target 6.1.

Inactivation of hepatitis A virus and murine norovirus on surfaces of plastic, steel and raspberries using steam-ultrasound treatment.

The leading causes of foodborne viral disease outbreaks are human norovirus and hepatitis A virus (HAV). Their environmental persistence enables contamination of kitchen surfaces and crops often consumed raw, such as berries. Many decontamination procedures are inefficient and unsuitable for surfaces of industrial kitchen environments and soft fruits. In this study, we investigated the efficiency of a novel surface decontamination technology, combining steam and ultrasound (steam-ultrasound). Plastic, steel or raspberry surfaces were spiked with the norovirus surrogate, murine norovirus (MNV), and HAV, and steam-ultrasound treated at 85, 90 and 95 °C for 0-5 s. Post treatment viruses were titrated for survival by plaque assay and for genome stability by real-time quantitative PCR (RT-qPCR) of nucleic acid extracts. Survival of viruses were estimated in a log-linear model and the treatment time requirements for each decimal reduction (D value) in viral survival were calculated. The estimated D values of MNV or HAV were 0.4-0.2 or 1.1-0.8 s on plastic, 0.9-0.7 or 1.4-0.8 s on steel and 1.6-1.7 or 3.2-4.7 s on raspberries. No clear trend of genome reduction was observed with tested treatment parameters. Raspberries treated up to 4 s retained its natural texture and visual appeal similar to untreated controls whilst monitored for 7 days. In conclusion, steam-ultrasound treatment can within seconds reduce the titre of foodborne viruses on surfaces of plastic, steel and raspberries. This may particularly benefit industrial scale production of soft fruits for raw consumption and for swift non-hazardous decontamination of industrial kitchen surfaces.

An Optimised Direct Lysis Method for Viral RNA Extraction and Detection of Foodborne Viruses on Fruits and Vegetables.

Detection of norovirus (NoV) and hepatitis A virus (HAV) on fruits and vegetables using current standard methodologies can be inefficient. Method optimisation focussing on ease, rapidity and increased viral RNA recovery is needed for efficient reverse transcription (RT)-qPCR detection of viruses. A simple and quick direct lysis method for RNA extraction was optimised (method A) to achieve increased viral RNA recovery and minimised RT-qPCR inhibition by increasing the volume of lysis buffer and inclusion of pectinase, Plant RNA Isolation Aid and OneStep PCR Inhibitor Removal Kit. Method A and an internal method structurally comparable to the ISO 15216 standard (method B) were compared for their efficiencies to recover viral RNA from the process controls, mengovirus (MC0) and murine norovirus (MNV), spiked in 13 types of fruits, vegetables, compound foods or seeds/nuts. All extracts (> 61) were also analysed for RT-qPCR inhibition and for natural contamination of NoV and HAV. The overall mean extraction efficiencies of MC0 and MNV were 36 ± 31 and 44 ± 38%, respectively, for method A and 9 ± 16 and 5 ± 11%, respectively, for method B. Inhibition of RT-qPCR amplification of RNA from NoV genogroup (G)I, NoV GII, and HAV ranged from 5 ± 10 to 13 ± 14% for method A and 34 ± 36 to 48 ± 40% for method B. NoV GII was detected in samples of strawberries and seaweed processed by both methods. In conclusion, the new direct lysis method showed an overall better performance compared to the modified ISO 15216 standard and should be validated for implementation in analysis of viruses in foods of plant origin.

Combining the Allosteric Inhibitor Asciminib with Ponatinib Suppresses Emergence of and Restores Efficacy against Highly Resistant BCR-ABL1 Mutants.

BCR-ABL1 point mutation-mediated resistance to tyrosine kinase inhibitor (TKI) therapy in Philadelphia chromosome-positive (Ph+) leukemia is effectively managed with several approved drugs, including ponatinib for BCR-ABL1T315I-mutant disease. However, therapy options are limited for patients with leukemic clones bearing multiple BCR-ABL1 mutations. Asciminib, an allosteric inhibitor targeting the myristoyl-binding pocket of BCR-ABL1, is active against most single mutants but ineffective against all tested compound mutants. We demonstrate that combining asciminib with ATP site TKIs enhances target inhibition and suppression of resistant outgrowth in Ph+ clinical isolates and cell lines. Inclusion of asciminib restores ponatinib's effectiveness against currently untreatable compound mutants at clinically achievable concentrations. Our findings support combining asciminib with ponatinib as a treatment strategy for this molecularly defined group of patients.

Inhibition of nSMase2 Reduces the Transfer of Oligomeric α-Synuclein Irrespective of Hypoxia.

Recently, extracellular vesicles (EVs), such as exosomes, have been proposed to play an influential role in the cell-to-cell spread of neurodegenerative diseases, including the intercellular transmission of α-synuclein (α-syn). However, the regulation of EV biogenesis and its relation to Parkinson's disease (PD) is only partially understood. The generation of EVs through the ESCRT-independent pathway depends on the hydrolysis of sphingomyelin by neutral sphingomyelinase 2 (nSMase2) to produce ceramide, which causes the membrane of endosomal multivesicular bodies to bud inward. nSMase2 is sensitive to oxidative stress, a common process in PD brains; however, little is known about the role of sphingomyelin metabolism in the pathogenesis of PD. This is the first study to show that inhibiting nSMase2 decreases the transfer of oligomeric aggregates of α-syn between neuron-like cells. Furthermore, it reduced the accumulation and aggregation of high-molecular-weight α-syn. Hypoxia, as a model of oxidative stress, reduced the levels of nSMase2, but not its enzymatic activity, and significantly altered the lipid composition of cells without affecting EV abundance or the transfer of α-syn. These data show that altering sphingolipids can mitigate the spread of α-syn, even under hypoxic conditions, potentially suppressing PD progression.

Genomic landscape of neutrophilic leukemias of ambiguous diagnosis.

Chronic neutrophilic leukemia (CNL), atypical chronic myeloid leukemia (aCML), and myelodysplastic/myeloproliferative neoplasms, unclassifiable (MDS/MPN-U) are a group of rare and heterogeneous myeloid disorders. There is strong morphologic resemblance among these distinct diagnostic entities as well as a lack of specific molecular markers and limited understanding of disease pathogenesis, which has made diagnosis challenging in certain cases. The treatment has remained empirical, resulting in dismal outcomes. We, therefore, performed whole-exome and RNA sequencing of these rare hematologic malignancies and present the most complete survey of the genomic landscape of these diseases to date. We observed a diversity of combinatorial mutational patterns that generally do not cluster within any one diagnosis. Gene expression analysis reveals enrichment, but not cosegregation, of clinical and genetic disease features with transcriptional clusters. In conclusion, these groups of diseases represent a continuum of related diseases rather than discrete diagnostic entities.

Targeting BCR-ABL1 in Chronic Myeloid Leukemia by PROTAC-Mediated Targeted Protein Degradation.

Although the use of ATP-competitive tyrosine kinase inhibitors of oncoprotein BCR-ABL1 has enabled durable responses in patients with chronic myeloid leukemia (CML), issues of drug resistance and residual leukemic stem cells remain. To test whether the degradation of BCR-ABL1 kinase could offer improved response, we developed a series of proteolysis-targeting chimera (PROTAC) that allosterically target BCR-ABL1 protein and recruit the E3 ligase Von Hippel-Lindau, resulting in ubiquitination and subsequent degradation of the oncogenic fusion protein. In both human CML K562 cells and murine Ba/F3 cells expressing BCR-ABL1, lead compound GMB-475 induced rapid proteasomal degradation and inhibition of downstream biomarkers, such as STAT5, and showed increased sensitivity compared with diastereomeric controls lacking degradation activity. Notably, GMB-475 inhibited the proliferation of certain clinically relevant BCR-ABL1 kinase domain point mutants and further sensitized Ba/F3 BCR-ABL1 cells to inhibition by imatinib, while demonstrating no toxicity toward Ba/F3 parental cells. Reverse phase protein array analysis suggested additional differences in levels of phosphorylated SHP2, GAB2, and SHC associated with BCR-ABL1 degradation. Importantly, GMB-475 reduced viability and increased apoptosis in primary CML CD34+ cells, with no effect on healthy CD34+ cells at identical concentrations. GMB-475 degraded BCR-ABL1 and reduced cell viability in primary CML stem cells. Together, these findings suggest that combined BCR-ABL1 kinase inhibition and protein degradation may represent a strategy to address BCR-ABL1-dependent drug resistance, and warrant further investigation into the eradication of persistent leukemic stem cells, which rely on neither the presence nor the activity of the BCR-ABL1 protein for survival. SIGNIFICANCE: Small-molecule-induced degradation of BCR-ABL1 in CML provides an advantage over inhibition and provides insights into CML stem cell biology. GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/79/18/4744/F1.large.jpg.

Clinical resistance to crenolanib in acute myeloid leukemia due to diverse molecular mechanisms.

FLT3 mutations are prevalent in AML patients and confer poor prognosis. Crenolanib, a potent type I pan-FLT3 inhibitor, is effective against both internal tandem duplications and resistance-conferring tyrosine kinase domain mutations. While crenolanib monotherapy has demonstrated clinical benefit in heavily pretreated relapsed/refractory AML patients, responses are transient and relapse eventually occurs. Here, to investigate the mechanisms of crenolanib resistance, we perform whole exome sequencing of AML patient samples before and after crenolanib treatment. Unlike other FLT3 inhibitors, crenolanib does not induce FLT3 secondary mutations, and mutations of the FLT3 gatekeeper residue are infrequent. Instead, mutations of NRAS and IDH2 arise, mostly as FLT3-independent subclones, while TET2 and IDH1 predominantly co-occur with FLT3-mutant clones and are enriched in crenolanib poor-responders. The remaining patients exhibit post-crenolanib expansion of mutations associated with epigenetic regulators, transcription factors, and cohesion factors, suggesting diverse genetic/epigenetic mechanisms of crenolanib resistance. Drug combinations in experimental models restore crenolanib sensitivity.

Metagenomic analysis of viruses in toilet waste from long distance flights-A new procedure for global infectious disease surveillance.

Human viral pathogens are a major public health threat. Reliable information that accurately describes and characterizes the global occurrence and transmission of human viruses is essential to support national and global priority setting, public health actions, and treatment decisions. However, large areas of the globe are currently without surveillance due to limited health care infrastructure and lack of international cooperation. We propose a novel surveillance strategy, using metagenomic analysis of toilet material from international air flights as a method for worldwide viral disease surveillance. The aim of this study was to design, implement, and evaluate a method for viral analysis of airplane toilet waste enabling simultaneous detection and quantification of a wide range of human viral pathogens. Toilet waste from 19 international airplanes was analyzed for viral content, using viral capture probes followed by high-throughput sequencing. Numerous human pathogens were detected including enteric and respiratory viruses. Several geographic trends were observed with samples originating from South Asia having significantly higher viral species richness as well as higher abundances of salivirus A, aichivirus A and enterovirus B, compared to samples originating from North Asia and North America. In addition, certain city specific trends were observed, including high numbers of rotaviruses in airplanes departing from Islamabad. Based on this study we believe that central sampling and analysis at international airports could be a useful supplement for global viral surveillance, valuable for outbreak detection and for guiding public health resources.