[The practical training of medical students in General Medicine: Important characteristics of teaching practices]. / Die praktische Ausbildung des medizinischen Nachwuchses ­ Identifizierung von Lehrpraxen-Charakteristika in der Allgemeinmedizin.

BACKGROUND: Comprehensive general medical care is endangered by the socio-demographic development of the population as well as by recruitment problems in general medicine. Medical students' interest in general medicine increases if they experience their internships positively. However, the characteristics of teaching practice important to medical students have not been systematically investigated. METHODS: We conducted four focus groups with a total of 22 students. The group discussions were recorded, transcribed, and evaluated by means of qualitative content analysis. RESULTS: Students who rated their internship positively also expressed greater interest in becoming a family doctor. Variables that mattered to students were: Instruction and supervision by the owner's teaching practice and feedback behavior, being treated with respect, opportunity to work independently, the broad spectrum of diseases and the varied daily work when working as a family doctor, long-standing doctor-patient relationships, and good work-life balance. Variables that decreased students' interest in becoming a family doctor were the economic risk of starting a business, and being self-employed, referring patients to specialists for "interesting findings" and a poor work-life balance. CONCLUSION: The practical training of medical students in teaching practices has the potential to increase students' interest in working as a family doctor. This requires motivated teaching practice owners, who give students a positive insight into everyday practice.

Biogenesis of a specialized plant-fungal interface during host cell internalization of Golovinomyces orontii haustoria.

Powdery mildew fungi are biotrophic pathogens that require living plant cells for their growth and reproduction. Elaboration of a specialized cell called a haustorium is essential for their pathogenesis, providing a portal into host cells for nutrient uptake and delivery of virulence effectors. Haustoria are enveloped by a modified plant plasma membrane, the extrahaustorial membrane (EHM), and an extrahaustorial matrix (EHMx), across which molecular exchange must occur, but the origin and composition of this interfacial zone remains obscure. Here we present a method for isolating Golovinomyces orontii haustoria from Arabidopsis leaves and an ultrastructural characterization of the haustorial interface. Haustoria were progressively encased by deposits of plant cell wall polymers, delivered by secretory vesicles and multivesicular bodies (MVBs) that ultimately become entrapped within the encasement. The EHM and EHMx were not labelled by antibodies recognizing eight plant cell wall and plasma membrane antigens. However, plant resistance protein RPW8.2 was specifically recruited to the EHMs of mature haustoria. Fungal cell wall-associated molecular patterns such as chitin and ß-1,3-glucans were exposed at the surface of haustoria. Fungal MVBs were abundant in haustoria and putative exosome vesicles were detected in the paramural space and EHMx, suggesting the existence of an exosome-mediated secretion pathway.

High-throughput confocal imaging of intact live tissue enables quantification of membrane trafficking in Arabidopsis.

Membrane compartmentalization and trafficking within and between cells is considered an essential cellular property of higher eukaryotes. We established a high-throughput imaging method suitable for the quantitative detection of membrane compartments at subcellular resolution in intact epidermal tissue. Whole Arabidopsis (Arabidopsis thaliana) cotyledon leaves were subjected to quantitative confocal laser microscopy using automated image acquisition, computational pattern recognition, and quantification of membrane compartments. This revealed that our method is sensitive and reliable to detect distinct endomembrane compartments. We applied quantitative confocal laser microscopy to a transgenic line expressing GFP-2xFYVE as a marker for endosomal compartments during biotic or abiotic stresses, and detected markedly quantitative adaptations in response to changing environments. Using a transgenic line expressing the plasma membrane-resident syntaxin GFP-PEN1, we quantified the pathogen-inducible extracellular accumulation of this fusion protein at fungal entry sites. Our protocol provides a platform to study the quantitative and dynamic changes of endomembrane trafficking, and potential adaptations of this machinery to physiological stress.