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Fruiting bodies are among the most complex multicellular structures formed by fungi, and the molecular mechanisms that regulate their development are far from understood. However, studies with a number of fungal model organisms have started to shed light on this developmental process. One of these model organisms is Sordaria macrospora, a filamentous ascomycete from the order Sordariales. This fungus has been a genetic model organism since the 1950s, but its career as a model organism for molecular genetics really took off in the 1990s, when the establishment of a transformation protocol, a mutant collection, and an indexed cosmid library provided the methods and resources to start revealing the molecular mechanisms of fruiting body development. In the 2000s, "omics" methods were added to the S. macrospora tool box, and by 2020, 58 developmental genes have been identified in this fungus. This review gives a brief overview of major method developments for S. macrospora, and then focuses on recent results characterizing different processes involved in regulating development including several regulatory protein complexes, autophagy, transcriptional and chromatin regulation, and RNA editing. KEY POINTS •Sordaria macrospora is a model system for analyzing fungal fruiting body development. •More than 100 developmental mutants are available for S. macrospora. •More than 50 developmental genes have been characterized in S. macrospora.The filamentous fungus Aspergillus terreus has been successfully used for industrial production of itaconic acid (IA) for many years. The IA biosynthesis pathway has recently been characterized at a molecular genetic level as an IA gene cluster by a clone-based transcriptomic approach. The cluster consists of four genes, including genes for cis-aconitic acid decarboxylase (cadA), a predicted transcription factor (tf), a mitochondrial organic acid transporter (mttA) and an MFS (major facilitator superfamily) type transporter (mfsA). In this research, we performed expressed sequence tag (EST) analysis and systematic gene deletions to further investigate the role of those genes during IA biosynthesis in A. pseudoterreus ATCC32359. EST analysis showed a similar expression pattern among those four genes that were distinct from neighboring genes and further confirmed that they belong to the same biosynthesis cluster. Systematic gene deletion analysis demonstrated that tf, cadA, mttA and mfsA genes in the cluster are essential for IA production; deletion of any of them will either completely abolish the IA production or dramatically decrease the amount of IA produced. selleck chemicals The tf gene plays a regulatory role in this cluster. Deletion of tf led to decreased expression levels of cadA, mttA and mfsA. More importantly, a significant amount of aconitic acid was detected in the cadA deletion strain but not in the other deletion strains. Therefore, by deleting only one gene, the cadA, we established a novel microbial host for the production of aconitic acid and other value-added chemicals from sugars in lignocellulosic biomass.2-Phenylethanol (2-PE) is an important flavor compound but also impairs cell growth severely, which in turn blocks its bioproduction. However, the molecular mechanism of 2-PE tolerance is unclear. In this study, a superb 2-PE stress-tolerant and producing yeast, Candida glycerinogenes, was selected to uncover the underlying mechanism of 2-PE tolerance. We discovered that Hap5 is an essential regulator to 2-PE resistance, and its induction by 2-PE stress occurs at the post-transcriptional level, rather than at the transcriptional level. Under 2-PE stress, Hap5 is activated and imported into the nucleus rapidly. Then, the nuclear Hap5 binds to the glutathione synthetase (gsh2) promoter via CCAAT box, to induce the expression of gsh2 gene. The increased gsh2 expression contributes to enhanced cellular glutathione content, and consequently alleviates ROS accumulation, lipid peroxidation, and cell membrane damage caused by 2-PE toxicity. Specifically, increasing the expression of gsh2 is effective in improving not just 2-PE tolerance (33.7% higher biomass under 29 mM 2-PE), but also 2-PE production (16.2% higher). This study extends our knowledge of 2-PE tolerance mechanism and also provides a promising strategy to improve 2-PE production.BACKGROUND Practical contents are gaining in importance in medical curricula in Germany. In times of a lack of applicants, practical courses provide an increased level of interest for students in the respective disciplines. A practical appeal of ophthalmology is microsurgical procedures. A microsurgical suture wet lab can be an introduction for medical students. OBJECTIVE Assessment of the increased interest in ophthalmology through evaluation of a suture wet lab course, including suturing under a microscope. MATERIAL AND METHODS The data were obtained from the practical course in ophthalmology during the sixth semester for medical students at the university medical center in Mainz in April 2019. In a questionnaire developed in collaboration with the Center for Quality Assurance and Development various statements had to be answered and evaluated using ordinal scales. RESULTS A total of 64 evaluation questionnaires from 8 groups each with 8 medical students supervised by different tutors were analyzed. The wet lab was rated with 1.24 ± 0.5 (mean ± SD) on a German school grade scale (1 = best, 6 = worst). There was agreement (1 = total agreement, 7 = total disagreement) concerning the desire for further microscopic wet labs (1.86 ± 1.28) as well as learning more ophthalmological surgical techniques (2.02 ± 1.13). The interest in ophthalmology (1 = very high interest, 7 = very low interest) increased from 3.66 ± 1.55 before the course to 2.52 ± 1.00. CONCLUSION Interest in ophthalmology can be increased through a microsurgical wet lab. The interest of students can therefore be awakened, which can have advantageous effects in job application situations and in research work. In this way experience and practical skills in ophthalmology can already be acquired during medical studies.Nodular scleritis and necrotizing scleritis are rare complications of acanthamoeba keratitis. This article presents the case of a 61-year-old female patient who had suffered from persistent keratitis in the right eye for more than 4 months. The patient was initially treated with propamidine isethionate and polyhexamethylene biguanide eye drops. A penetrating limbo-keratoplasty was performed. Examination of the corneal explant showed acanthamoeba cysts. In the following 5 months the sclera showed recurrent abscesses. A total of two thermal cauterizations and three amniotic membrane grafts were carried out. To our knowledge, this is the first case of sclerokeratitis after acanthamoeba keratitis which was treated with a combination of thermal cauterization and amniotic membrane transplantation. Further studies are necessary to investigate this procedure as an alternative to the established cryotherapy.In this work, an electrochemiluminescence (ECL) biosensor was fabricated for the selective detection of vascular endothelial growth factor (VEGF165). g-C3N4/PDDA/CdSe nanocomposites were used as the ECL substrate. Then, DNA labeled at the 5' end with amino groups (DNA1) was immobilized on the surface of g-C3N4/PDDA/CdSe nanocomposite-modified glassy carbon electrode (GCE) by amido linkage. AuNP-labeled target DNA (Au-DNA2) could hybridize with DNA1 to form a double strand. The ECL of the g-C3N4/PDDA/CdSe nanocomposite was efficiently quenched due to the resonance energy transfer between CdSe QDs and Au NPs. After VEGF165 was recognized and bound by Au-DNA2, the double helix was disrupted, and the energy transfer was broken. In this case, Au-DNA2 was released from the electrode surface, and the ECL intensity recovered to a higher level. Under optimal conditions, this ECL biosensor possesses excellent selectivity, accuracy, and stability for VEGF165 detection in a linear range of 2 pg mL-1 to 2 ng mL-1 with a detection limit of 0.68 pg mL-1. In addition, this assay has been successfully applied to the determination of VEGF165 in serum samples. Graphical abstract Schematic representation of the electrochemiluminescence sensor based on a g-C3N4/PDDA/CdSe nanocomposite, which can be determined in the concentration of vascular endothelial growth factor in serum.A novel electrochemical sensor, platinum nanoparticles/graphene nanoplatelets/multi-walled carbon nanotubes/β-cyclodextrin composite (PtNPs-GNPs-MWCNTs-β-CD) modified carbon glass electrode (GCE), was fabricated and used for the sensitive detection of folic acid (FA). The PtNPs-GNPs-MWCNTs-β-CD nanocomposite was easily prepared with an ultrasound-assisted assembly method, and it was characterized by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The electrochemical behavior of FA at PtNPs-GNPs-MWCNTs-β-CD/GCE was investigated in detail. Some key experimental parameters such as pH, amount of PtNPs-GNPs-MWCNTs-β-CD composite, and scan rate were optimized. A good linear relationship (R2 = 0.9942) between peak current of cyclic voltammetry (CV) and FA concentration in the range 0.02-0.50 mmol L-1 was observed at PtNPs-GNPs-MWCNTs-β-CD/GCE. The detection limit was 0.48 μmol L-1 (signal-to-noise ratio = 3). A recovery of 97.55-102.96% was obtained for the determination of FA in FA pills (containing 0.4 mg FA per pill) at PtNPs-GNPs-MWCNTs-β-CD/GCE, indicating that the modified electrode possessed relatively high sensitivity and stability for the determination of FA in real samples.A rapid, simple, and sensitive technique for the quantitative detection of fluoxetine and norfluoxetine enantiomers in biological fluids was developed based on the combination of field-amplified sample stacking (FASS)-related capillary electrophoresis (CE) with ultrasound-assisted dispersive liquid-liquid microextraction (UA-DLLME). The extraction efficiency of UA-DLLME was strongly related to extraction time, salt concentration, type of extraction and dispersion solvents, and volume of extraction and dispersion solvents. The extracted fluoxetine and norfluoxetine enantiomers in a mixture of 50% methanol and 50% deionized water were efficiently stacked using FASS and then separated using cyclodextrin-modified CE. Under optimal conditions of FASS (chiral selector, 3 mM trimethyl-β-cyclodextrin; and background electrolyte, 100 mM phosphate buffer) and UA-DLLME (extraction solvent, 200 μL of acetone; and dispersed solvent, 50 μL of C2H2Cl4 in 1 mL of the sample solution), the obtained enrichment factors of fluoxetine and norfluoxetine enantiomers reached approximately 2000. The linear ranges for the quantification of fluoxetine and norfluoxetine enantiomers were 0.3-150 and 0.6-150 nM, respectively. The relative standard deviations in peak areas and migration time for four analytes were less than 3.3% and 6.3%, respectively. The proposed system provided limits of detection (signal-to-noise ratio of 3) for four analytes corresponding to 0.1 nM. The precision and accuracy for urine and serum samples were less than 6.8 and 8.3%, respectively. These findings suggested that the proposed system exhibited a high potential for the reliable determination of fluoxetine and norfluoxetine enantiomers in clinical samples. Graphical abstract.