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.

Functional genomic landscape of acute myeloid leukaemia.

The implementation of targeted therapies for acute myeloid leukaemia (AML) has been challenging because of the complex mutational patterns within and across patients as well as a dearth of pharmacologic agents for most mutational events. Here we report initial findings from the Beat AML programme on a cohort of 672 tumour specimens collected from 562 patients. We assessed these specimens using whole-exome sequencing, RNA sequencing and analyses of ex vivo drug sensitivity. Our data reveal mutational events that have not previously been detected in AML. We show that the response to drugs is associated with mutational status, including instances of drug sensitivity that are specific to combinatorial mutational events. Integration with RNA sequencing also revealed gene expression signatures, which predict a role for specific gene networks in the drug response. Collectively, we have generated a dataset-accessible through the Beat AML data viewer (Vizome)-that can be leveraged to address clinical, genomic, transcriptomic and functional analyses of the biology of AML.

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.

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.

Reduced retromer function results in the accumulation of amyloid-beta oligomers.

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by a progressive loss of multiple cognitive functions. Accumulation of amyloid beta oligomers (oAß) play a major role in the neurotoxicity associated with the disease process. One of the early affected brain regions is the hippocampus, wherein a reduction of the vacuolar protein sorting-associated protein 35 (VPS35), the core protein comprising the retromer complex involved in cellular cargo sorting, has been identified. To investigate the role of the retromer function on the accumulation and clearance of oAß, we reduced retromer function by selectively inhibiting VPS35 gene expression using siRNA in differentiated neuronal SH-SY5Y cells. As cell-to-cell transfer of oAß to new brain regions is believed to be important for disease progression we investigated the effect of VPS35 reduction both in cells with direct uptake of oAß and in cells receiving oAß from donor cells. We demonstrate that reduced retromer function increases oAß accumulation in both cell systems, both the number of cells containing intracellular oAß and the amount within them. This effect was shown at different time points and regardless if the oAß originated from the extracellular milieu or via a direct neuronal cell-to-cell transfer. Interestingly, not only did reduced VPS35 cause oAß accumulation, but oAß treatment alone also lead to a reduction of VPS35 protein content. The accumulated oAß seems to co-localize with VPS35 and early endosome markers. Together, these findings provide evidence that reduced retromer function decreases the ability for neurons to transport and clear neurotoxic oAß received through different routes resulting in the accumulation of oAß. Thus, enhancing retromer function may be a potential therapeutic strategy to slow down the pathophysiology associated with the progression of AD.

Alzheimer's disease pathology propagation by exosomes containing toxic amyloid-beta oligomers.

The gradual deterioration of cognitive functions in Alzheimer's disease is paralleled by a hierarchical progression of amyloid-beta and tau brain pathology. Recent findings indicate that toxic oligomers of amyloid-beta may cause propagation of pathology in a prion-like manner, although the underlying mechanisms are incompletely understood. Here we show that small extracellular vesicles, exosomes, from Alzheimer patients' brains contain increased levels of amyloid-beta oligomers and can act as vehicles for the neuron-to-neuron transfer of such toxic species in recipient neurons in culture. Moreover, blocking the formation, secretion or uptake of exosomes was found to reduce both the spread of oligomers and the related toxicity. Taken together, our results imply that exosomes are centrally involved in Alzheimer's disease and that they could serve as targets for development of new diagnostic and therapeutic principles.

Sources of Calicivirus contamination in foodborne outbreaks in Denmark, 2005-2011--the role of the asymptomatic food handler.

BACKGROUND: Norovirus (NoV) is the predominant cause of foodborne disease outbreaks. Virus contamination may occur during all steps of food processing, from production to preparation and serving. The relative importance of these different routes of contamination is unknown. METHODS: The purpose of this study was to estimate the proportions of outbreaks caused by asymptomatic and symptomatic food handlers (FHs). Reports of foodborne NoV and sapovirus outbreaks (n=191) that occurred over a 7-year period were extracted, reviewed, and categorized according to the available evidence for source of contamination. RESULTS: In 64 (34%) of the outbreaks, contamination from FHs took place during preparation or serving of food. In the majority of these outbreaks (n=41; 64%), the FHs were asymptomatic during food handling. Some had been in contact with ill household members before handling the food and remained asymptomatic; others developed symptoms shortly after or were post-symptomatic. In 51 (27%) of the outbreaks, contamination occurred during production of the food, and in 55 (29%) of the outbreaks, contamination had supposedly occurred after serving a guest at a self-serve buffet. CONCLUSIONS: Guidelines regarding exclusion of FHs where household members suffer from gastroenteritis could limit the number of outbreaks.

Presence of Pathogenic Bacteria and Viruses in the Daycare Environment.

The number of children in daycare centers (DCCs) is rising. This increases exposure to microorganisms and infectious diseases. Little is known about which bacteria and viruses are present in the DCC environment and where they are located. In the study described in this article, the authors set out to determine the prevalence of pathogenic bacteria and viruses and to find the most contaminated fomites in DCCs. Fifteen locations in each DCC were sampled for bacteria, respiratory viruses, and gastrointestinal viruses. The locations were in the toilet, kitchen, and playroom areas and included nursery pillows, toys, and tables, among other things. Coliform bacteria were primarily found in the toilet and kitchen areas whereas nasopharyngeal bacteria were found mostly on toys and fabric surfaces in the playroom. Respiratory viruses were omnipresent in the DCC environment, especially on the toys.

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.

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.

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.

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.

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.

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.

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.

Caenorhabditis elegans intersectin: a synaptic protein regulating neurotransmission.

Intersectin is a multifunctional protein that interacts with components of the endocytic and exocytic pathways, and it is also involved in the control of actin dynamics. Drosophila intersectin is required for viability, synaptic development, and synaptic vesicle recycling. Here, we report the characterization of intersectin function in Caenorhabditis elegans. Nematode intersectin (ITSN-1) is expressed in the nervous system, and it is enriched in presynaptic regions. The C. elegans intersectin gene (itsn-1) is nonessential for viability. In addition, itsn-1-null worms do not display any evident phenotype, under physiological conditions. However, they display aldicarb-hypersensitivity, compatible with a negative regulatory role of ITSN-1 on neurotransmission. ITSN-1 physically interacts with dynamin and EHS-1, two proteins involved in synaptic vesicle recycling. We have previously shown that EHS-1 is a positive modulator of synaptic vesicle recycling in the nematode, likely through modulation of dynamin or dynamin-controlled pathways. Here, we show that ITSN-1 and EHS-1 have opposite effects on aldicarb sensitivity, and on dynamin-dependent phenotypes. Thus, the sum of our results identifies dynamin, or a dynamin-controlled pathway, as a potential target for the negative regulatory role of ITSN-1.

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.

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.