Neurologic complications of breast cancer.

Breast cancer is a heterogeneous disease with unique neurologic complications that can arise from central nervous system (CNS) involvement or secondary to treatments themselves. As progress is made, with more targeted therapies and combinations available, particularly in the realm of human epidermal growth factor receptor 2 (HER2)-positive disease, the role of these new agents in patients with CNS disease is gradually evolving, although intracranial efficacy itself is lagging. At the same time, both systemic and local standard therapies pose clinical challenges regarding neurologic complications, such as peripheral neuropathy and cognitive changes. The development of new agents, such as immunotherapy, and new strategies, such as incorporating systemic therapies into local therapy, unveil new presentations of neurological complications.

A Single-Arm, Open-Label Phase II Study of ONC201 in Recurrent/Refractory Metastatic Breast Cancer and Advanced Endometrial Carcinoma.

BACKGROUND: ONC201 is a small molecule that can cause nonapoptotic cell death through loss of mitochondrial function. Results from the phase I/II trials of ONC201 in patients with refractory solid tumors demonstrated tumor responses and prolonged stable disease in some patients. METHODS: This single-arm, open-label, phase II clinical trial evaluated the efficacy of ONC201 at the recommended phase II dose (RP2D) in patients with recurrent or refractory metastatic breast or endometrial cancer. Fresh tissue biopsies and blood were collected at baseline and at cycle 2 day 2 for correlative studies. RESULTS: Twenty-two patients were enrolled; 10 patients with endometrial cancer, 7 patients with hormone receptor-positive breast cancer, and 5 patients with triple-negative breast cancer. The overall response rate was 0%, and the clinical benefit rate, defined by complete response (CR) + partial response (PR) + stable disease (SD), was 27% (n = 3/11). All patients experienced an adverse event (AE), which was primarily low grade. Grade 3 AEs occurred in 4 patients; no grade 4 AEs occurred. Tumor biopsies did not show that ONC201 consistently induced mitochondrial damage or alterations in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) or the TRAIL death receptors. ONC201 treatment caused alterations in peripheral immune cell subsets. CONCLUSION: ONC201 monotherapy did not induce objective responses in recurrent or refractory metastatic breast or endometrial cancer at the RP2D dose of 625 mg weekly but had an acceptable safety profile (ClinicalTrials.gov Identifier: NCT03394027).

First application of loop-mediated isothermal amplification (LAMP) assays for rapid identification of mating type in the heterothallic fungus Aspergillus fumigatus.

BACKGROUND: Loop-mediated isothermal amplification (LAMP) assays, which operate at a single temperature and require no postreaction processing, have been described for rapid species-specific detection of numerous fungi. The technology has much less commonly been applied to identification of other key genetic traits such as fungicide resistance, and has not yet been applied to mating-type determination in any fungus. OBJECTIVES: To develop first LAMP assays for mating-type identification in a fungus, in this instance with the saprophytic mould and human opportunistic pathogen Aspergillus fumigatus, a heterothallic ascomycete requiring isolates of opposite mating type (MAT1-1, MAT1-2) for sexual reproduction. METHODS: New LAMP primer sets, targeted to MAT gene sequences, were screened against 34 A fumigatus isolates (of known mating type) from diverse clinical, environmental and geographic sources to establish whether they could distinguish MAT1-1 or MAT1-2 genotypes. RESULTS AND CONCLUSIONS: The new assays, operating at a single temperature of 65°C, correctly identified the mating type of A fumigatus isolates in <20 minutes, and thus have numerous research and practical applications. Similar MAT LAMP assays could now be developed for other fungi of agricultural, environmental, industrial and/or medical importance.

Simulated consultations: a sociolinguistic perspective.

BACKGROUND: Assessment of consulting skills using simulated patients is widespread in medical education. Most research into such assessment is sited in a statistical paradigm that focuses on psychometric properties or replicability of such tests. Equally important, but less researched, is the question of how far consultations with simulated patients reflect real clinical encounters--for which sociolinguistics, defined as the study of language in its socio-cultural context, provides a helpful analytic lens. DISCUSSION: In this debate article, we draw on a detailed empirical study of assessed role-plays, involving sociolinguistic analysis of talk in OSCE interactions. We consider critically the evidence for the simulated consultation (a) as a proxy for the real; (b) as performance; (c) as a context for assessing talk; and (d) as potentially disadvantaging candidates trained overseas. Talk is always a performance in context, especially in professional situations (such as the consultation) and institutional ones (the assessment of professional skills and competence). Candidates who can handle the social and linguistic complexities of the artificial context of assessed role-plays score highly--yet what is being assessed is not real professional communication, but the ability to voice a credible appearance of such communication. Fidelity may not be the primary objective of simulation for medical training, where it enables the practising of skills. However the linguistic problems and differences that arise from interacting in artificial settings are of considerable importance in assessment, where we must be sure that the exam construct adequately embodies the skills expected for real-life practice. The reproducibility of assessed simulations should not be confused with their validity. Sociolinguistic analysis of simulations in various professional contexts has identified evidence for the gap between real interactions and assessed role-plays. The contextual conditions of the simulated consultation both expect and reward a particular interactional style. Whilst simulation undoubtedly has a place in formative learning for professional communication, the simulated consultation may distort assessment of professional communication These sociolinguistic findings contribute to the on-going critique of simulations in high-stakes assessments and indicate that further research, which steps outside psychometric approaches, is necessary.

Storytelling and affiliation between healthcare staff in Schwartz Round interactions: A conversation analytic study.

It is well known that the demands of working in healthcare can take a psychological toll on staff. Schwartz Centre Rounds are an intervention aimed at supporting staff wellbeing through providing a forum to talk about the emotional, social and ethical complexities of such work, employing facilitated storytelling and group discussion to try and achieve this. However, while prior research, through extensive interviews and surveys, has found Schwartz Rounds to be effective in fostering compassion and wellbeing amongst participants, the talk that occurs within Schwartz Rounds themselves has not been explored. One mechanism that has been considered in how Schwartz Rounds function is the creation of a 'counter-cultural', conversational space, suggesting the nature of the interactions themselves may be important in achieving their beneficial effects. Using conversation analytic (CA) methods, we examine Schwartz Rounds in the UK to address, at a detailed micro-level, how sequences of talk work to accomplish the key aims of this setting. Five separate one-hour Schwartz Rounds were recorded across three UK hospital Trusts, between January 2019 and February 2020. Our analysis addresses how panellists tell their stories in a way that emphasises the uniqueness of their experience but also provides a generalisable emotional 'upshot' and 'stance' for the audience to later respond to. We then focus in on how audience members are able to respond to these stories affiliatively, offering endorsements, generalisations and second stories. Drawing on prior CA literature examining support groups and psychotherapy, we consider how the format of Schwarz Rounds creates important opportunities for interpersonal affiliation in this context. Considering these interactional features alongside other research findings on Schwartz Rounds, we discuss how opportunities for interactional affiliation may be central to their success, with implications for how these interactions can be best facilitated.

Ten-Fold Elevation of Chromogranin A Level Unrelated to a Neuroendocrine Tumor: A Case Report of the Diagnostic Interference of Proton Pump Inhibitors.

Chromogranin A (CgA) is a well-known biomarker for neuroendocrine tumors (NETs). However, due to its non-specificity, a proper assessment of CgA test results requires a detailed knowledge of the factors, conditions, and medications influencing its serum concentration. We describe a case of a 61-year-old patient presenting with a mass suspicious of a gastrointestinal NET and an exceedingly high level of serum CgA persistent after mass resection. Following a thorough review of patient's medical history and clinical presentation, along with radiographic and pathological findings, no evidence of a NET was detected. A trial of proton-pump inhibitor (PPI) withdrawal led to a dramatic normalization of CgA level, marking it as the culprit causing this tumor marker elevation. This case highlights the significant impact of PPI use on CgA level, and should incentivize clinicians to provide proper education to patients prior to testing.

Epidemiological Studies of Pan-Azole Resistant Aspergillus fumigatus Populations Sampled during Tulip Cultivation Show Clonal Expansion with Acquisition of Multi-Fungicide Resistance as Potential Driver.

Pan-azole resistant isolates are found in clinical and environmental Aspergillus fumigatus (Af) populations. Azole resistance can evolve in both settings, with Af directly targeted by antifungals in patients and, in the environment, Af unintendedly exposed to fungicides used for material preservation and plant disease control. Resistance to non-azole fungicides, including methyl benzimidazole carbamates (MBCs), quinone outside inhibitors (QoIs) and succinate dehydrogenase inhibitors (SDHIs), has recently been reported. These fungicide groups are not used in medicine but can play an important role in the further spread of pan-azole resistant genotypes. We investigated the multi-fungicide resistance status and the genetic diversity of Af populations sampled from tulip field soils, tulip peel waste and flower compost heaps using fungicide sensitivity testing and a range of genotyping tools, including STRAf typing and sequencing of fungicide resistant alleles. Two major clones were present in the tulip bulb population. Comparisons with clinical isolates and literature data revealed that several common clonal lineages of TR34/L98H and TR46/Y121F/T289A strains that have expanded successfully in the environment have also acquired resistance to MBC, QoI and/or SDHI fungicides. Strains carrying multiple fungicide resistant alleles have a competitive advantage in environments where residues of multiple fungicides belonging to different modes of action are present.

The Multi-Fungicide Resistance Status of Aspergillus fumigatus Populations in Arable Soils and the Wider European Environment.

The evolution and spread of pan-azole resistance alleles in clinical and environmental isolates of Aspergillus fumigatus is a global human health concern. The identification of hotspots for azole resistance development in the wider environment can inform optimal measures to counteract further spread by minimizing exposure to azole fungicides and reducing inoculum build-up and pathogen dispersal. We investigated the fungicide sensitivity status of soil populations sampled from arable crops and the wider environment and compared these with urban airborne populations. Low levels of azole resistance were observed for isolates carrying the CYP51A variant F46Y/M172V/E427K, all belonging to a cluster of related cell surface protein (CSP) types which included t07, t08, t13, t15, t19, and t02B, a new allele. High levels of resistance were found in soil isolates carrying CYP51A variants TR34/L98H and TR46/Y121F/T289A, all belonging to CSP types t01, t02, t04B, or t11. TR46/Y121F/M172V/T289A/G448S (CSP t01) and TR46/Y121F/T289A/S363P/I364V/G448S (CSP t01), a new haplotype associated with high levels of resistance, were isolated from Dutch urban air samples, indicating azole resistance evolution is ongoing. Based on low numbers of pan-azole resistant isolates and lack of new genotypes in soils of fungicide-treated commercial and experimental wheat crops, we consider arable crop production as a coldspot for azole resistance development, in contrast to previously reported flower bulb waste heaps. This study also shows that, in addition to azole resistance, several lineages of A. fumigatus carrying TR-based CYP51A variants have also developed acquired resistance to methyl benzimidazole carbamate, quinone outside inhibitor and succinate dehydrogenase (Sdh) inhibitor fungicides through target-site alterations in the corresponding fungicide target proteins; beta-tubulin (F200Y), cytochrome b (G143A), and Sdh subunit B (H270Y and H270R), respectively. Molecular typing showed that several multi-fungicide resistant strains found in agricultural soils in this study were clonal as identical isolates have been found earlier in the environment and/or in patients. Further research on the spread of different fungicide-resistant alleles from the wider environment to patients and vice versa can inform optimal practices to tackle the further spread of antifungal resistance in A. fumigatus populations and to safeguard the efficacy of azoles for future treatment of invasive aspergillosis.

Chemotherapy and Beyond: Infections in the Era of Old and New Treatments for Hematologic Malignancies.

Treatment options for hematologic malignancies have been rapidly expanding in the past decade, resulting in better survival outcomes for many patients. Infection is an important cause of morbidity and mortality in this patient population. Cytotoxic chemotherapy has well-studied infectious risks related to the degree and duration of myelosuppression. Targeted therapies and immunotherapies have less clearly predictable infectious risk and diverse effects on immune function. This review discusses contemporary management of hematologic malignancies, followed by special discussion of novel agents, including signaling/small molecule inhibitors, monoclonal antibodies, immunomodulators, and immunotherapies, for treatment of hematologic malignancies with focus on infectious risk.

Analysing voice quality and pitch in interactions of emergency care simulation.

Background/aims: In emergency care, healthcare professionals (HCPs) interact with both a patient and their colleagues at the same time. How HCPs regulate the two distinct interactions is our central interest. Focusing on HCPs' use of their voice quality and pitch, a multimodal analysis of the interaction in a simulation training session was conducted. Our aims are (1) to compare the use of HCPs' voice quality and pitch in HCP-patient and HCP-HCP interactions, (2) to examine how different voice quality and pitch function in interaction, and (3) to develop the research methodology so as to integrate multimodal features in emergency care interaction for analysis. Methods: Three HCPs performed a scripted acute care scenario (chest pain) at the simulation centre. The multimodal corpus-based approach was applied to analyse the varying voice pitch and quality of the HCPs, in interactions with a simulated patient (SP) and with two other HCPs, in emergency care training. Results: The HCPs tended to use a clear voice when they talk to an SP and a 'shattered' voice to colleagues in the team. The pitch was raised to talk to an SP, by Helen (a nurse) and Mike (a doctor). Conclusion: This indicates that the HCPs strategically change their voice quality and pitch according to the addressees, regulating the interaction.

Sociolinguistic factors affecting performance in the Clinical Skills Assessment of the MRCGP: a mixed-methods approach.

BACKGROUND: Differential performance in clinical skills assessments is a widespread phenomenon, for which there remain few explanations. AIM: To better understand the conversational contexts of simulated consultations and how candidates actually behave in these consultations and to determine sociolinguistic factors for high- and low-performing candidates. DESIGN & SETTING: Taking the Membership of the Royal College of General Practitioners' (MRCGP) clinical skills assessment (CSA) examination as a model, this research applied sociolinguistic analyses to case videos of 198 consecutive candidates presenting for the CSA examination. METHOD: Using a mixed-methods approach, both quantitative and qualitative sociolinguistics methodologies were combined to analyse video consultations, and findings were compared with those from group discussions with MRCGP examiners. RESULTS: There is more 'talk' in simulated consultations than in real life. On macroanalysis, there was little difference between poor- and well-performing candidates. However, microanalysis found subtle differences in structuring consultations, metacommunication, picking up cues, and misunderstandings with and giving explanations to patients. Formulaic talk, contrary to examiners' perceptions was more common in successful candidates, but it was personalised and sited appropriately in the consultation. CONCLUSION: This is an interactionally demanding form of clinical assessment, that requires giving support to candidates and a more analytic approach to the development of interpersonal skills. Sociolinguistic features of consulting to help trainers and candidates prepare for the CSA are identified.

Simulated patients versus manikins in acute-care scenarios.

BACKGROUND: Manikins and simulated patients (SPs) are commonly used in health care education and assessment. SPs appear to offer a more realistic experience for learners than 'plastic' manikins, and might be expected to engender interactions that approximate real clinical practice more closely. The analyses of linguistic patterns and touch are methodologies that could be used to explore this hypothesis. Our research aims were: (1) to compare verbal interactions and the use of procedural touch by health care workers (HCWs) in scenarios with SPs and with manikins; and (2) to evaluate the methodologies used to inform a large-scale study. METHODS: We conducted a pilot preliminary comparative study on conversations and touch in two scenarios in emergency care training, each performed using an SP and a high-fidelity manikin. The setting was a simulation centre. Two scripted acute-care scenarios (chest pain and acute breathlessness) were performed using both an SP and a high-fidelity manikin. Audiovisual recordings were subsequently analysed. Two teams comprising a clinician (medical student), a senior nurse and a clinical support worker took part in the study. The linguistics research methodology integrated a multimodal corpus-based approach and discourse analysis. Simulated patients appear to offer a more realistic experience for learners than 'plastic' manikins RESULTS: Overall, HCWs had far more interactions with the SPs than with the manikins, and the utterances and behaviours were more akin to what one would expect in real clinical practice. HCWs used procedural touch without the patient's permission much more frequently with the manikin. DISCUSSION: These methodologies offer opportunities for further research projects studying interactions with SPs, manikins and real clinical practice.

An Intervention to Promote Sleep and Reduce ADHD Symptoms.

OBJECTIVE: To determine the effect of a sleep hygiene education module and prescriptive sleep routine for children ages 5 through 11 years with attention deficit hyperactivity disorder (ADHD). METHODS: The 20-week pilot project used a pre-/posttest design to assess sleep and ADHD symptoms. The Child Sleep Habits Questionnaire (CHSQ) and Vanderbilt Assessment Scale-Parent Form survey were provided to assess sleep and ADHD behaviors of participants at baseline and 6 weeks after implementation of the sleep hygiene routine. Fifty-three children participated in the project. Of these, 23 scored 42 or greater on the CHSQ, indicating a sleep disorder, and received the intervention. RESULTS: The CHSQ and Vanderbilt scores indicated a significant improvement in sleep quality and reduction in ADHD symptoms after implementation of the sleep hygiene routine (CHSQ: p < .001, d = .928; Vanderbilt Questions 1-9: p < .001, d = .473; Vanderbilt Questions 10-18: p = .004; d = .329). CONCLUSION: A provider-instructed sleep hygiene routine in children with ADHD improves sleep quality and reduces ADHD symptoms.

Analysis of mammalian gene function through broad-based phenotypic screens across a consortium of mouse clinics.

The function of the majority of genes in the mouse and human genomes remains unknown. The mouse embryonic stem cell knockout resource provides a basis for the characterization of relationships between genes and phenotypes. The EUMODIC consortium developed and validated robust methodologies for the broad-based phenotyping of knockouts through a pipeline comprising 20 disease-oriented platforms. We developed new statistical methods for pipeline design and data analysis aimed at detecting reproducible phenotypes with high power. We acquired phenotype data from 449 mutant alleles, representing 320 unique genes, of which half had no previous functional annotation. We captured data from over 27,000 mice, finding that 83% of the mutant lines are phenodeviant, with 65% demonstrating pleiotropy. Surprisingly, we found significant differences in phenotype annotation according to zygosity. New phenotypes were uncovered for many genes with previously unknown function, providing a powerful basis for hypothesis generation and further investigation in diverse systems.

Simultaneous real-time PCR detection of Fusarium asiaticum, F. ussurianum and F. vorosii, representing the Asian clade of the F. graminearum species complex.

Due to the repeated discovery of new members of the Fusarium graminearum species complex (FGSC), some of the F. graminearum sensu stricto (s.s.)-specific qPCR assays developed to date have since been shown to be non-specific. In this study, a probe-based qPCR method was developed, targeting a sterol 14-alpha demethylase (CYP51) paralogue, CYP51C unique to the genus Fusarium, for the simultaneous detection of F. asiaticum, F. ussurianum and F. vorosii. Specificity of the assay was demonstrated for a wide range of Fusarium species, including all tested FGSC members (n=6), originating from different hosts and geographic regions. Alongside a previously published assay for detection of F. graminearum, we were able to show that members of the Asian clade of the FGSC (i.e. F. asiaticum, F. ussurianum and F. vorosii) were the primary etiological agent in wheat seeds samples originating from Central-East China. The grain samples from the UK tested negative for the presence of the FGSC's Asian clade and positive for presence of F. graminearum. It is likely that only F. graminearum s.s. is present in the UK, but the presence of other FGSC members cannot be ruled out and need further investigation.

A comparative phenotypic and genomic analysis of C57BL/6J and C57BL/6N mouse strains.

BACKGROUND: The mouse inbred line C57BL/6J is widely used in mouse genetics and its genome has been incorporated into many genetic reference populations. More recently large initiatives such as the International Knockout Mouse Consortium (IKMC) are using the C57BL/6N mouse strain to generate null alleles for all mouse genes. Hence both strains are now widely used in mouse genetics studies. Here we perform a comprehensive genomic and phenotypic analysis of the two strains to identify differences that may influence their underlying genetic mechanisms. RESULTS: We undertake genome sequence comparisons of C57BL/6J and C57BL/6N to identify SNPs, indels and structural variants, with a focus on identifying all coding variants. We annotate 34 SNPs and 2 indels that distinguish C57BL/6J and C57BL/6N coding sequences, as well as 15 structural variants that overlap a gene. In parallel we assess the comparative phenotypes of the two inbred lines utilizing the EMPReSSslim phenotyping pipeline, a broad based assessment encompassing diverse biological systems. We perform additional secondary phenotyping assessments to explore other phenotype domains and to elaborate phenotype differences identified in the primary assessment. We uncover significant phenotypic differences between the two lines, replicated across multiple centers, in a number of physiological, biochemical and behavioral systems. CONCLUSIONS: Comparison of C57BL/6J and C57BL/6N demonstrates a range of phenotypic differences that have the potential to impact upon penetrance and expressivity of mutational effects in these strains. Moreover, the sequence variants we identify provide a set of candidate genes for the phenotypic differences observed between the two strains.

Overexpression of the sterol 14α-demethylase gene (MgCYP51) in Mycosphaerella graminicola isolates confers a novel azole fungicide sensitivity phenotype.

BACKGROUND: The recent evolution towards resistance to azole fungicides in European populations of the wheat pathogen Mycosphaerella graminicola has been caused by the progressive accumulation of mutations in MgCYP51 gene, encoding the azole target sterol 14α-demethylase. Particular combinations of mutations have been shown specifically to affect the interaction of the MgCYP51 protein with different members of the azole class. Although additional mechanisms, including increased MgCYP51 expression and enhanced active efflux, have been proposed, the genetic changes underlying these mechanisms are unknown. RESULTS: Analysis of the azole sensitivities of recent M. graminicola isolates identified a novel phenotype, seemingly independent of changes in MgCYP51 coding sequence. Characterised by a 7-16-fold reduction in in vitro sensitivity to all azoles tested and by growth on seedlings at higher doses of azoles in glasshouse tests compared with isolates carrying the same MgCYP51 variant (L50S, S188N, I381V, ΔY459/G460, N513K), isolates with this phenotype constitutively overexpress MgCYP51 by between 10- and 40-fold compared with the wild type. Analysis of sequences upstream of the predicted MgCYP51 translation start codon identified a novel 120 bp indel, considered to be an insertion, in isolates overexpressing MgCYP51. CONCLUSIONS: The identification of an insertion in the predicted MgCYP51 promoter in azole-resistant isolates overexpressing MgCYP51 is the first report of a genetic mechanism, other than changes in target-site coding sequence, affecting sensitivity to multiple azoles in field isolates of M. graminicola. The identification of recent isolates overexpressing MgCYP51 confirms the ongoing evolution and diversification of resistance mechanisms in European populations of M. graminicola.

Risk assessment studies on succinate dehydrogenase inhibitors, the new weapons in the battle to control Septoria leaf blotch in wheat.

Chemical control of Septoria leaf blotch, caused by Mycosphaerella graminicola, is essential to ensure wheat yield and food security in most European countries. Mycosphaerella graminicola has developed resistance to several classes of fungicide and, with the efficacy of azoles gradually declining over time, new modes of action and/or improvements in host varietal resistance are urgently needed to ensure future sustainable disease control. Several new-generation carboxamide fungicides with broad-spectrum activity have recently been introduced into the cereal market. Carboxamides inhibit succinate dehydrogenase (Sdh) of the mitochondrial respiratory chain (complex II) but, because of their single-site specificity, these fungicides may be prone to resistance development. The objective of this study was to assess the risk of resistance development to different Sdh inhibitor (SDHI) fungicides in M. graminicola. UV mutagenesis was conducted to obtain a library of carboxin-resistant mutants. A range of SDHI resistance-conferring mutations was found in Sdh subunits B, C and D. Pathogenicity studies with a range of Sdh variants did not detect any fitness costs associated with these mutations. Most of the amino acid residues identified (e.g. B-S221P/T, B-H267F/L/N/Y, B-I269V and D-D129E/G/T) are directly involved in forming the cavity in which SDHI fungicides bind. Docking studies of SDHI fungicides in structural models of wild-type and mutated Sdh complexes also indicated which residues were important for the binding of different SDHI fungicides and showed a different binding for fluopyram. The predictive power of the model was also shown. Further diagnostic development, enabling the detection of resistant alleles at low frequencies, and cross-resistance studies will aid the implementation of anti-resistance strategies to prolong the cost-effectiveness and lifetime of SDHI fungicides.

The use of a CYP51C gene based PCR-RFLP assay for simultaneous detection and identification of Fusarium avenaceum and F. tricinctum in wheat.

Contamination of cereals with mycotoxins such as beauvericin (BEA), enniatins (Ens) and moniliformin (MON) is mainly caused by Fusarium avenaceum and F. tricinctum. This is a world-wide problem which requires rapid and sensitive detection methods. To allow for high throughput screening of large numbers of samples, a diagnostic PCR method was developed for the simultaneous detection of F. avenaceum and F. tricinctum. The interspecific divergence found in the Fusarium-specific CYP51C gene was used to design species-specific PCR primers. The specificity of the assay was demonstrated for DNA samples extracted from a wide range of Fusarium species belonging to the Fusarium head blight (FHB) complex, as well as for naturally-infected grain samples. The PCR-amplified products were digested with the restriction enzyme XbaI to enable differentiation between F. avenaceum and F. tricinctum. This PCR- restriction fragment length polymorphism (RFLP) assay proved to be a simple and relatively inexpensive method highly suited for routine detection and identification of F. avenaceum and F. tricinctum in wheat samples.