Novel Multiplex and Loop-Mediated Isothermal Amplification Assays for Rapid Species and Mating-Type Identification of Oculimacula acuformis and O. yallundae (Causal Agents of Cereal Eyespot), and Application for Detection of Ascospore Dispersal and In Planta Use.

Eyespot, caused by the related fungal pathogens Oculimacula acuformis and O. yallundae, is an important cereal stem-base disease in temperate parts of the world. Both species are dispersed mainly by splash-dispersed conidia but are also known to undergo sexual reproduction, yielding apothecia containing ascospores. Field diagnosis of eyespot can be challenging, with other pathogens causing similar symptoms, which complicates eyespot management strategies. Differences between O. acuformis and O. yallundae (e.g., host pathogenicity and fungicide sensitivity) require that both be targeted for effective disease management. Here, we develop and apply two molecular methods for species-specific and mating-type (MAT1-1 or MAT1-2) discrimination of O. acuformis and O. yallundae isolates. First, a multiplex PCR-based diagnostic assay targeting the MAT idiomorph region was developed, allowing simultaneous determination of both species and mating type. This multiplex PCR assay was successfully applied to type a global collection of isolates. Second, the development of loop-mediated isothermal amplification (LAMP) assays targeting ß-tubulin sequences, which allow fast (<9 min) species-specific discrimination of global O. acuformis and O. yallundae isolates, is described. The LAMP assay can detect very small amounts of target DNA (1 pg) and was successfully applied in planta. In addition, mating-type-specific LAMP assays were also developed for rapid (<12 min) genotyping of O. acuformis and O. yallundae isolates. Finally, the multiplex PCR-based diagnostic was applied, in conjunction with spore trapping in field experiments, to provide evidence of the wind dispersal of ascospores from a diseased crop. The results indicate an important role of the sexual cycle in the dispersal of eyespot.

Dental dilemmas: Endodontics or dental implants?

UNLABELLED: This narrative review explores treatment planning options in restorative dentistry. The growth of dental implants, as an accessible and predictable treatment option, gives practitioners a useful tool for managing the missing tooth or teeth with a hopeless prognosis. Traditionally, endodontics and fixed prosthodontics have been used to restore teeth and spaces where the outlook for such treatment appears reasonable. Practitioners may, however, question the predictability and cost effectiveness of such an approach where, at times, it might appear that replacement of a compromised tooth with a dental implant could be a more predictable option. The evidence base for these treatment options is explored and discussed, and suggestions are made for future management strategies. CLINICAL RELEVANCE: A clear knowledge and understanding of the scientific literature for implants and endodontic treatment is necessary if practitioners are to make an evidence-based approach when treatment planning these modalities for their patients. This is particularly true in cases where there may appear to be a reasonable choice between the two of these.

A guide to the appraisal of research: 1. The nature of research.

UNLABELLED: Evidence-based dentistry has grown in recent years. The oceans of journals and research papers can be difficult to navigate. Gauging the quality of such publications and, more importantly, their clinical relevance may be challenging.This narrative review explores the nature and relevance of dental research to clinical dentistry and provides simple guidelines for critical appraisal of such research. In this respect, all dental clinical practice should be underpinned, where possible, with evidence that treatments carried out are contemporary and effective. CLINICAL RELEVANCE: A working knowledge of dental research and its relevance to clinical practice should allow practitioners to select best evidence to inform their clinical practice.

A guide to the appraisal of research: 2. What's in a number?

UNLABELLED: This review explores the nature of numbers in dental research, their importance and, at a basic level, how these are handled. This includes the important numbers that practitioners should look at when appraising research and discussion of some commonly found statistical terms. CLINICAL RELEVANCE: In order to apply evidence-based dentistry in clinical practice, dentists should have an understanding of the basic numbers reported in dental research, and an appreciation of this should allow them to identify which papers are most valuable to inform their clinical practice.

Trends in indirect dentistry: 2. post and core restorations.

Modern post preparation techniques reflect a change from what was once considered a prosthodontic procedure to one which marries endodontic principles with sound understanding of mechanical objectives. This article presents a rationale for techniques in materials selection and post-space preparation. A review of literature is presented and practical clinical guidelines are suggested when post placement is considered the treatment of choice.

Trends in indirect dentistry: 1. Case selection.

Indirect restorations can provide a conservative, functional and aesthetic method for managing a patient's dentition. These restorations, however, are only as good as the environment in which they are placed. Many restorations, though technically satisfactory, may fail because assessment and subsequent treatment planning has been unsatisfactory. This article presents a philosophy of case selection mindful of factors related to the patient, operator and oral environment. The wrong treatment, however skilfully executed, may invite failure.