The quality of traditional sMDT(single-centered multi-disciplinary treatment team) teaching model will be affected by the centerlocation, leve of heathcare service, performance habits, dominant ...disciplines, and superior technologies, which will directly affect the depth and breadth of students' knowledge learning. Based on the traditional sMDT teaching, the new multi-centered MDT (mMDT) teaching model establishes cooperative teaching relationships with multiple hospitals and disciplines, cocultivates undergraduate and graduate students, and regularly conducts mMDT teaching, which effectively avoids the defects of the traditional sMDT teaching model and meets the needs of students at different levels. Through the mMDT teaching mode, students can learn the advanced clinical reasoning and techniques in different disciplines or hospitals, have a deeper understanding of the concept of comprehensive diagnosis and treatment of tumors, and improve the comprehensive quality of clinical research.
The invasive ascomycete
Hymenoscyphus fraxineus
has been threatening
Fraxinus excelsior
populations throughout Europe for over two decades. Since the infection and first colonization by the pathogen ...occurs in leaves, leaf-colonizing microorganisms have been discussed as a barrier and as possible biocontrol agents against the disease. To identify fungal groups with health-supporting potential, we compared the fungal microbiota of compound leaves from susceptible and tolerant ash trees in four ash stands with high
H. fraxineus
exposure. The fungal communities were analyzed both culture-independently by ITS2 amplicon sequencing and by the taxonomic classification of 1,704 isolates using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) or sequencing of the entire ITS region. The fungal community structure did not show significant differences depending on the health status. However, for several OTUs and a MALDI group, a significantly higher abundance was found in tolerant ash trees. Thus, the yeast
Papiliotrema flavescens
was significantly increased and accounted for 12.3% of the mycobiome of tolerant ashes (OTU0003), and it had also a distinctly higher abundance among the isolates. The filamentous ascomycete
Sarocladium strictum
was increased 24-fold among the isolates of tolerant trees, but its abundance was comparably low. An
in vitro
screening for the growth inhibition of the pathogen
via
cocultivation resulted in 28 yeast-like isolates and 79 filamentous fungi with antagonistic activity. A statistical cocultivation test on two
H. fraxineus
strains confirmed six of the yeast-like isolates that suppressed
H. fraxineus
significantly, from 39–50%, two of them through a fungicidal effect. The highest inhibition rates among the yeasts were found for three isolates belonging to
Aureobasidium pullulans
and
P. flavescens
. The cocultivation test of the filamentous isolates revealed higher effects compared to the yeasts. Four isolates showed significant inhibition of both
H. fraxineus
strains with a rate of 72–100%, and five further isolates inhibited only one
H. fraxineus
strain significantly. The most effective isolates were members of the genus
Cladosporium
. During the next step,
in planta
tests will be necessary to verify the efficacy of the antagonistic isolates and to assess their suitability as biocontrol agents.
Conventional hydroponic units producing disease free plants with 30–40% faster growth rate are regarded as a promising agricultural alternative in case of unfertile/metal contaminated soil. However, ...most nutrients in hydroponics medium remain unused and are drained off creating environmental pollution. To reduce the nutrient load and eutrophication effects, cocultivation of plant and microalgae in the hydroponic units has attracted the attention of researchers over the past few years. The present study aims to explore the influence of initial inoculum concentration of a native algal consortium over the performance of hydroponic system. Cocultivation of tomato plant with varying initial inoculum algal concentration of 0.2–0.8 mg/ml, showed that 0.8 mg/ml concentration resulted in positive interactions between microalgae and plant, with algal and plant productivity of 0.149 ± 0.024 and 0.328 ± 0.087 g/m2/d respectively after 42 days. Higher chlorophyll accumulation, along with nutritionally rich algal and plant biomass revealed lack of unwanted competition during cocultivation. Increase in dissolved oxygen during cocultivation was corroborated with efficient root respiration. Highly developed roots also provided adequate metabolic energy thus significantly increasing the nutrient uptake and accumulation, thereby reducing the nutrient load during drainage. The supernatant after algal harvesting (recycled media) supported plant growth for 24 days due to limited nutrients. Such system with microalgae symbiotically favouring plant productivity and simultaneous nutrient load reduction would ultimately support in attaining sustainable agriculture.
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•Mixed algal consortium and plants showed symbiotic interaction during coculture.•Initial microalgal inoculum influences the performance of cocultivation in hydroponics.•Dissolved oxygen level in hydroponics was higher due to microalgal cultivation.•Higher algal and plant productivity was obtained with 0.8 mg/ml of algal inoculum.•85% of N, P and K uptake efficiency was achieved during hydroponic cocultivation.
Fluorescence-activated droplet sorting (FADS) combined with transcription factor (TF)-based biosensors allows for the high-throughput isolation of clones that overproduce extracellular metabolites. ...Here, we developed a FADS-TF coculture pipeline for improving erythritol production in Yarrowia lipolytica. The pipeline comprises three droplet operation steps: the generation and incubation of droplets that encapsulate yeast mutants with single-cell resolution, the pico-injection of fluorescence-based erythritol-biosensing Escherichia coli, and the sorting of the highest fluorescence droplets with an approximately 1 ‰ survival ratio. With the picodroplet-based workflow, we initially carried out a two-stage temperature and pH control strategy on the droplets to separate the erythritol production and detection processes. We then eliminated the background expression of biosensors by utilizing pH-sensitive erythromycin, finally establishing a complete version of the coculture screening system. The optimal strain, Yarrowia lipolytica S4-9, contributed 231.2 g/L erythritol after a 114-h fermentation and was successfully screened from the fourth round of the mutant library. Compared to the parent, the erythritol yield and production rate of Yarrowia lipolytica S4-9 increased by 16.97 % and 26.09 %, respectively, in a 5-L bioreactor. Our results indicated that FADS-TFs have great potential for the high-throughput screening of extracellular products as additional biosensors become available.
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•The microfluidic-based platform enabled cocultivation of biosensors and producers.•A novel FADS workflow enabled the high-throughput screening of Yarrowia lipolytica.•The optimal mutant strain that dramatically increased erythritol production was selected.
Since the onset of microbiology in the late 19th century, scientists have been growing microorganisms almost exclusively as pure cultures, resulting in a limited and biased view of the microbial ...world. Only a paradigm shift in cultivation techniques – from axenic to mixed cultures – can allow a full comprehension of the (chemical) communication of microorganisms, with profound consequences for natural product discovery, microbial ecology, symbiosis, and pathogenesis, to name a few areas. Three main technical advances during the last decade are fueling the realization of this revolution in microbiology: microfluidics, next-generation 3D-bioprinting, and single-cell metabolomics. These technological advances can be implemented for large-scale, systematic cocultivation studies involving three or more microorganisms. In this review, we present recent trends in microbiology tools and discuss how these can be employed to decode the chemical language that microorganisms use to communicate.
A limited, biased, and anthropocentric view of the microbial world with focus on fast-growing copiotrophic species has emerged from classical axenic cultivation approaches.
Recent (meta)genomic insights unveiled the potential hidden in microbial diversity. However, cultivation-independent approaches cannot replace cultivation techniques. Cultivation techniques have to evolve further – from axenic to mixed cultures – to fully understand the microbial world.
Newly emerged tools, including microfluidics, bioprinting, high-throughput screening, and single-cell analytics, need to be fully implemented and integrated with existing (microbiology) techniques to systematically investigate and exploit microbial cocultures.
Samples of cell suspension represents seven different densities were exposed to three electrotreatment 40, 50, 60 volt and for a period of time 50, 100 msec then the densities were mixed with ...different concentration of Ri plasmid 25, 50, 75 Microliter. The electrotreatment samples cultured by embedding in to agar drops in solid-liquid cultures.
Results proved that the electrotreatment enhanced division of cell suspension and caused a clear increase of microcolonies, the total number of colonies reached 3276 when suspension culture was exposed to 50v / 100 msec., these colonies developed to microcalli and the percentage was 90%. The Callus developed from these colonies was light green in color and friable in texture.
O-Methylated phenylpropanoids, which are generally present in small amounts in plants, have improved or distinct biological activities and pharmacological properties as opposed to their unmethylated ...counterparts. Although microbial production could be a useful tool for the efficient and environment-friendly production of methylated phenylpropanoids, a high-yield microbial production of neither tri-methylated stilbenes nor di-/tri-methylated flavonoids has been achieved to date.
A methyltransferase from Streptomyces avermitilis (SaOMT2), which has been known to possess 7-O-methylation activity toward several flavonoids, exhibited more diverse regiospecificity and catalyzed mono-, di-, and tri-methylation of stilbene, flavanone, and flavone when it was expressed in Streptomyces venezuelae. For the efficient production of multi-methylated phenylpropanoids, a cocultivation system was developed by employing engineered Escherichia coli strains producing pterostilbene, naringenin, and apigenin, respectively, along with SaOMT2-expressing S. venezuelae mutant. Consequently, high-yield microbial production of tri-methylated stilbenes and di-/tri-methylated flavonoids (including 3,5,4'-trimethoxystilbene, 5-hydroxy-7,4'-dimethoxyflavanone, 4'-hydroxy-5,7-dimethoxyflavanone, 5,7,4'-trimethoxyflavanone, 5-hydroxy-7,4'-dimethoxyflavone, and 5,7,4'-trimethoxyflavone) has been demonstrated for the first time.
This cocultivation system based on the phenylpropanoid-producing E. coli and SaOMT2-expressing S. venezuelae provides an efficient tool for producing scarce and potentially valuable multi-methylated phenylpropanoids and will enable further development of these compounds as pharmaceuticals and nutraceuticals.
Lacto-N-tetraose (LNT) is an important neutral human milk oligosaccharide (HMO) and acts as a significant core structure for complex HMO biosynthesis. We previously achieved high-yield LNT ...biosynthesis (57.5 g/L) using fed-batch fermentation; however, residual lacto-N-triose II (LNTri II) was also found (21.58 g/L). Here, we re-engineered an efficient LNT-producing Escherichia coli with low LNTri II accumulation using genetically stable LNTri II-producing strains with a genomic insertion of lgtA (encoding β1,3-N-acetylglucosaminyltransferase). Comparable and low titers of LNT (3.73–4.61 g/L) and LNTri II (0.33–0.63 g/L), respectively, were obtained by introducing β1,3-galactosyltransferase. To reduce residual LNTri II, the E. coli transporter gene setA was disrupted, obviously reducing the accumulation of LNTri II and LNT. Next, the gene encoding β-N-acetylhexosaminidase (BbhI) was introduced into LNT-producing strains or E. coli BL21(DE3) for single- or mixed-strain cultivation, respectively. Finally, LNT was obtained (30.13 g/L) in a cocultivation system of mixed engineered strains without undesired LNTri II.
Cocultivation of the high cytochalasan-producing fungi Aspergillus flavipes and Chaetomium globosum resulted in the isolation of 11 undescribed Chae-type cytochalasans. Their structures were ...determined by spectroscopic data and NMR data calculations. Asperchaetoglobin A (1) was the first Chae-type cytochalasan possessing an unprecedented nitrogen bridge between C-17 and C-20 to generate a surprising 5/6/12/5 multiple ring system; asperchaetoglobins B and C (2 and 3) displayed higher oxidation with an additional epoxide at the thirteen-member ring; asperchaetoglobin D (4) was the second Chae-type cytochalasin featuring a 5/6/12 tricyclic ring system. The cytotoxic activities against five human cancer cell lines and antibacterial activities against Staphylococcus aureus and Colon bacillus of selected compounds were evaluated in vitro.
11 undescribed Chae-type cytochalasans were isolated from coculture of Aspergillus flavipes and Chaetomium globosum. Display omitted
•This is the first report of coculture of Aspergillus flavipes and Chaetomium globosum in liquid medium.•Eleven undescribed Chae-type cytochalasans asperchaetoglobins A-K were isolated and characterized.•Asperchaetoglobin A (1) possessing an unprecedented nitrogen bridge between C-17 and C-20.•The cytotoxic and antibacterial activities were evaluated in vitro.
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