The small secreted effector protein MiSSP7.6 of Laccaria bicolor is required for the establishment of ectomycorrhizal symbiosis

To establish and maintain a symbiotic relationship, the ectomycorrhizal fungus Laccaria bicolor releases mycorrhiza-induced small secreted proteins (MiSSPs) into host roots. Here, we have functionally characterized the MYCORRHIZA-iNDUCED SMALL SECRETED PROTEIN OF 7.6 kDa (MiSSP7.6) from L. bicolor by assessing its induced expression in ectomycorrhizae, silencing its expression by RNAi, and tracking in planta subcellular localization of its protein product. We also carried out yeast two-hybrid assays and bimolecular fluorescence complementation analysis to identify possible protein targets of the MiSSP7.6 effector in Populus roots. We showed that MiSSP7.6 expression is upregulated in ectomycorrhizal rootlets and associated extramatrical mycelium during the late stage of symbiosis development. RNAi mutants with a decreased MiSSP7.6 expression have a lower mycorrhization rate, suggesting a key role in the establishment of the symbiosis with plants. MiSSP7.6 is secreted, and it localizes both to the nuclei and cytoplasm in plant cells. MiSSP7.6 protein was shown to interact with two Populus Trihelix transcription factors. Furthermore, when coexpressed with one of the Trihelix transcription factors, MiSSP7.6 is localized to plant nuclei only. Our data suggest that MiSSP7.6 is a novel secreted symbiotic effector and is a potential determinant for ectomycorrhiza formation.Fil: Kang, Heng. Institut National de la Recherche Agronomique; FranciaFil: Chen, Xin. Institut National de la Recherche Agronomique; FranciaFil: Kemppainen, Minna Johanna. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Instituto de Microbiologia Basica y Aplicada. Laboratorio de Micologia Molecular.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pardo, Alejandro Guillermo. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Instituto de Microbiologia Basica y Aplicada. Laboratorio de Micologia Molecular.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Veneault Fourrey, Claire. Institut National de la Recherche Agronomique; FranciaFil: Kohler, Annegret. Institut National de la Recherche Agronomique; FranciaFil: Martin, Francis M.. Institut National de la Recherche Agronomique; Franci

The small secreted effector protein MiSSP7.6 of Laccaria bicolor is required for the establishment of ectomycorrhizal symbiosis

To establish and maintain a symbiotic relationship, the ectomycorrhizal fungus Laccaria bicolor releases mycorrhiza-induced small secreted proteins (MiSSPs) into host roots. Here, we have functionally characterized the MYCORRHIZA-iNDUCED SMALL SECRETED PROTEIN OF 7.6 kDa (MiSSP7.6) from L. bicolor by assessing its induced expression in ectomycorrhizae, silencing its expression by RNAi, and tracking in planta subcellular localization of its protein product. We also carried out yeast two-hybrid assays and bimolecular fluorescence complementation analysis to identify possible protein targets of the MiSSP7.6 effector in Populus roots. We showed that MiSSP7.6 expression is upregulated in ectomycorrhizal rootlets and associated extramatrical mycelium during the late stage of symbiosis development. RNAi mutants with a decreased MiSSP7.6 expression have a lower mycorrhization rate, suggesting a key role in the establishment of the symbiosis with plants. MiSSP7.6 is secreted, and it localizes both to the nuclei and cytoplasm in plant cells. MiSSP7.6 protein was shown to interact with two Populus Trihelix transcription factors. Furthermore, when coexpressed with one of the Trihelix transcription factors, MiSSP7.6 is localized to plant nuclei only. Our data suggest that MiSSP7.6 is a novel secreted symbiotic effector and is a potential determinant for ectomycorrhiza formation.Fil: Kang, Heng. Institut National de la Recherche Agronomique; FranciaFil: Chen, Xin. Institut National de la Recherche Agronomique; FranciaFil: Kemppainen, Minna Johanna. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Instituto de Microbiologia Basica y Aplicada. Laboratorio de Micologia Molecular.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pardo, Alejandro Guillermo. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Instituto de Microbiologia Basica y Aplicada. Laboratorio de Micologia Molecular.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Veneault Fourrey, Claire. Institut National de la Recherche Agronomique; FranciaFil: Kohler, Annegret. Institut National de la Recherche Agronomique; FranciaFil: Martin, Francis M.. Institut National de la Recherche Agronomique; Franci

A Standardized, Pragmatic Approach to Knee Ultrasound for Clinical Research in Osteoarthritis: The Johnston County Osteoarthritis Project

OBJECTIVE: This study sought to develop and employ a comprehensive and standardized ultrasound (US) protocol and scoring atlas for the evaluation of features relevant to knee osteoarthritis (KOA) in a community-based cohort in the United States, with the goals of demonstrating feasibility, reliability, and validity. METHODS: We utilized data from the fourth follow-up (2016-2018) of the Johnston County OA Project, which includes individuals with (~50%) and without radiographic KOA. All participants underwent standardized knee radiography and completed standard questionnaires including the Knee Injury and Osteoarthritis Outcome Score (KOOS). Bilateral knee US images were obtained by a trained sonographer using a standardized protocol and scored by trained rheumatologists using an atlas developed for this study. A total of 396 knees were each scored by two readers according to the atlas. Associations between US features, radiographic findings (graded by an expert radiologist), and KOOS scores were assessed. RESULTS: Overall interreader reliability for US scoring was fair to moderate. The strongest correlations between US and radiographic features were seen for osteophytes, and similarly strong correlations were seen between US osteophytes and overall radiographic Kellgren-Lawrence Grade, demonstrating criterion validity. Features of effusion/synovitis and osteophytes were most associated with KOOS pain and impaired function. CONCLUSION: US is a feasible, reliable, and valid method to assess features relevant to KOA in clinical and research settings. The protocol and atlas developed in this study can be utilized to evaluate KOA in a standardized fashion in future clinical studies, enabling greater utilization of this valuable modality in osteoarthritis

Pilot Point-of-Care Ultrasound Curriculum at Harvard Medical School: Early Experience

Introduction: Point-of-care ultrasound (POCUS) is expanding across all medical specialties. As the benefits of US technology are becoming apparent, efforts to integrate US into pre-clinical medical education are growing. Our objective was to describe our process of integrating POCUS as an educational tool into the medical school curriculum and how such efforts are perceived by students. Methods: This was a pilot study to introduce ultrasonography into the Harvard Medical School curriculum to first- and second-year medical students. Didactic and hands-on sessions were introduced to first-year students during gross anatomy and to second-year students in the physical exam course. Student-perceived attitudes, understanding, and knowledge of US, and its applications to learning the physical exam, were measured by a post-assessment survey. Results: All first-year anatomy students (n=176) participated in small group hands-on US sessions. In the second-year physical diagnosis course, 38 students participated in four sessions. All students (91%) agreed or strongly agreed that additional US teaching should be incorporated throughout the four-year medical school curriculum. Conclusion: POCUS can effectively be integrated into the existing medical school curriculum by using didactic and small group hands-on sessions. Medical students perceived US training as valuable in understanding human anatomy and in learning physical exam skills. This innovative program demonstrates US as an additional learning modality. Future goals include expanding on this work to incorporate US education into all four years of medical school

Promoting dual careers at higher education institutions: 31 benefits ranked by the project Student Athletes Erasmus+ Mobility in Europe (SAMEurope)

IntroductionThe project Student Athletes Erasmus+ Mobility in Europe identified and defined a total of 31 benefits offered to dual-career student athletes who are combining their university studies with high-level training. The project was co-funded by the European Union and carried out by five universities: Chalmers University of Technology (Sweden), Institut national des sciences appliquées de Lyon (France), University of Jyväskylä (Finland), Karlsruhe Institute of Technology (Germany), and Universitat Jaume I (Spain).MethodsThe purpose of the study was to rank these benefits by combining the perspectives of the university staff and the student athletes from each university in the consortium. The university staff included experts from sports services and the international relations office. A questionnaire was also sent to the dual-career athletes enrolled at the consortium's universities. Of the 514 dual-career athletes, 208 (116 women) completed the questionnaire. The overall response rate was 40.47%. The university staff assessed the importance of each benefit, how easy it was to implement at the institution, and whether or not the university offered the benefit to its students. The dual-career students rated each of the 31 benefits and indicated whether or not they had used them. A specific methodology was designed to rank these benefits using the ratings of the university staff and the student athletes. Intra-group and inter-group Pearson correlations were performed.ResultsThe results show a strong and significant correlation between the benefits from the perspective of the university staff (r = 0.710, p = 7.76E-7) and from the perspective of the students (r = 0.715, p = 2.44E-6). The correlation is moderate and significant when the benefits are correlated from the perspective of the two groups as a whole (0.363, p = 0.045), with the three most important benefits being the free use of sports facilities, justification for absences, and the adaptation of the pace of studies.DiscussionThe study makes visible the commitment of higher education institutions to facilitating the dual career of student athletes and identifies those benefits that may be of greater interest to European universities as a whole. The European perspective has been considered, while respecting the specificities of each university and the country in which it is located

The 2023 ACR/EULAR Classification Criteria for Calcium Pyrophosphate Deposition Disease

ObjectiveCalcium pyrophosphate deposition (CPPD) disease is prevalent and has diverse presentations, but there are no validated classification criteria for this symptomatic arthritis. The American College of Rheumatology (ACR) and EULAR have developed the first-ever validated classification criteria for symptomatic CPPD disease.MethodsSupported by the ACR and EULAR, a multinational group of investigators followed established methodology to develop these disease classification criteria. The group generated lists of candidate items and refined their definitions, collected de-identified patient profiles, evaluated strengths of associations between candidate items and CPPD disease, developed a classification criteria framework, and used multi-criterion decision analysis to define criteria weights and a classification threshold score. The criteria were validated in an independent cohort.ResultsAmong patients with joint pain, swelling, or tenderness (entry criterion) whose symptoms are not fully explained by an alternative disease (exclusion criterion), the presence of crowned dens syndrome or calcium pyrophosphate crystals in synovial fluid are sufficient to classify a patient as having CPPD disease. In the absence of these findings, a score >56 points using weighted criteria, comprising clinical features, associated metabolic disorders, and results of laboratory and imaging investigations, can be used to classify as CPPD disease. These criteria had a sensitivity of 92.2% and specificity of 87.9% in the derivation cohort (190 CPPD cases, 148 mimickers), whereas sensitivity was 99.2% and specificity was 92.5% in the validation cohort (251 CPPD cases, 162 mimickers).ConclusionThe 2023 ACR/EULAR CPPD disease classification criteria have excellent performance characteristics and will facilitate research in this field

In-person versus virtual musculoskeletal ultrasound education during the coronavirus disease 2019 pandemic: A single-center study

Background: Musculoskeletal ultrasound (MSKUS) curricula in physical medicine and rehabilitation (PM and R) residency programs have traditionally relied on in-person teaching workshops. However, the social distancing requirements during the coronavirus disease 2019 pandemic has led to a growing interest in remote ultrasound education. Aims: The aim of the study is to assess residents' MSKUS skill confidence after in-person vs. virtual MSKUS teaching workshops in a PM and R residency program. Materials and Methods: Twenty-one PM and R residents at a single academic center were assigned to either an in-person MSKUS demonstration by a sports medicine faculty member (n = 10) or a real-time virtual demonstration (n = 11) for workshops of the knee and wrist and hand. Surveys using Likert scales were employed to assess the change in residents' confidence with performing MSKUS examinations and their satisfaction with the curriculum. Results: There was a significant increase in confidence scores from baseline to postworkshop for both the virtual and in-person groups. The linear mixed effect model showed that there were no significant differences between the virtual and in-person groups for their baseline, postworkshop, and changes in confidence scores. All participants were satisfied with the updated curriculum. Conclusions: With the ongoing pandemic, incorporating virtual teaching platforms into existing MSKUS curricula is feasible and may provide similar improvements in residents' skill confidence as the in-person workshops. Successful implementation requires consideration of several factors, including ultrasound machine availability, participants' familiarity with technology, and the quality of the virtual streaming platform. A multi-institutional randomized controlled trial can further evaluate the effectiveness of remote MSKUS education for PM and R trainees

Long‐term dementia risk prediction by the LIBRA score: A 30‐year follow‐up of the CAIDE study

Objective As no causal treatment for dementia is available yet, the focus of dementia research is slowly shifting towards prevention strategies. Therefore, this study aimed to examine the predictive accuracy of the "LIfestyle for BRAin Health" (LIBRA) score, a weighted compound score of 12 modifiable risk and protective factors, for dementia and mild cognitive impairment (MCI) in midlife and late-life, and in individuals with high or low genetic risk based on presence of the apolipoprotein (APOE) epsilon 4 allele. Methods The LIBRA score was calculated for participants from the Finnish Cardiovascular Risk Factors, Aging and Dementia (CAIDE) population-based study examined in midlife (n = 1024) and twice in late-life (n = 604) up to 30 years later. Diagnoses of MCI and dementia were made according to established criteria. Cox proportional hazards models were used to assess the association between LIBRA and risk of dementia and MCI in models adjusted for sex and education (age as timescale). Results Higher midlife LIBRA scores were related to higher risk of dementia (hazard ratio [HR] = 1.27; 95% confidence interval [CI], 1.13-1.43) and MCI (unadjusted model: HR = 1.12; 95% CI, 1.03-1.22) up to 30 years later. Higher late-life LIBRA scores were related to higher risk of MCI (HR = 1.11; 95% CI, 1.00-1.25), but not dementia (HR = 1.02; 95% CI, 0.84-1.24). Higher late-life LIBRA scores were related to higher dementia risk among apolipoprotein E (APOE) epsilon 4 non-carriers. Conclusions Findings emphasize the importance of modifiable risk and protective factors for dementia prevention