Correction: Zhang et al. Bimodal Imaging of Tumors via Genetically Engineered Escherichia coli. Pharmaceutics 2022, 14, 1804.

In the original publication [...].

Bimodal Imaging of Tumors via Genetically Engineered Escherichia coli.

Although there are emerging innovations of molecular imaging probes to detect and image tumors, most of these molecular dyes and nanoparticles have limitations of low targetability in tumors and fast clearance when administered systemically. In contrast, some bacteria, such as Escherichia coli MG1655, can selectively proliferate in a hypoxic environment inside of a tumor for several days, which highlights the potential for the development of a genetically encoded multimodal imaging probe to monitor the progress of the tumor. Here, we developed bimodal imaging tumor-homing bacteria (GVs-miRFP680 MG1655) that allow both optical and acoustic imaging in tumor-bearing mice. An in vivo optical image system and a Vevo 2100 imaging system were applied to detect different imaging properties of the engineered bacteria in vivo. Our results show that the GVs-miRFP680 MG1655 bacteria can effectively integrate the advantages of low tissue absorbance from near-infrared fluorescent proteins and non-invasiveness from gas vesicles. We successfully developed GVs-miRFP680 MG1655 bacteria, which have both acoustic and optical imaging abilities in vitro and in vivo. The acoustic signal can last for up to 25 min, while the near-infrared fluorescence signal can last for up to 96 h. The combination of different imaging modalities in the tumor-homing bacteria may contribute to the non-invasive monitoring of the therapeutic effect of bacterial therapy in the future.

An evolutionarily stable strategy to colonize spatially extended habitats.

The ability of a species to colonize newly available habitats is crucial to its overall fitness1-3. In general, motility and fast expansion are expected to be beneficial for colonization and hence for the fitness of an organism4-7. Here we apply an evolution protocol to investigate phenotypical requirements for colonizing habitats of different sizes during range expansion by chemotaxing bacteria8. Contrary to the intuitive expectation that faster is better, we show that there is an optimal expansion speed for a given habitat size. Our analysis showed that this effect arises from interactions among pioneering cells at the front of the expanding population, and revealed a simple, evolutionarily stable strategy for colonizing a habitat of a specific size: to expand at a speed given by the product of the growth rate and the habitat size. These results illustrate stability-to-invasion as a powerful principle for the selection of phenotypes in complex ecological processes.

Effect of light quality on growth rate, carbohydrate accumulation, fatty acid profile and lutein biosynthesis of Chlorella sp. AE10.

Microalgae are feedstocks for multiple product development based on algal biorefinery concept. The effects of light quality (white, red and blue light emitting diodes) and macro-element starvations on Chlorella sp. AE10 were investigated under 20% CO2 and 850 µmol m-2 d-1. Nitrogen and phosphorus starvations had negative effects on its growth rate. The biomass productivities were decreased from day 1 and the highest one was 1.90 g L-1 d-1 under white light conditions. Phosphorus starvation promoted carbohydrate accumulation under three LED light sources conditions and the highest carbohydrate content was 75.9% using red light. Blue light increased lutein content to 9.58 mg g-1. The content of saturated fatty acids was significantly increased from 37.51% under blue light and full culture medium conditions to 77.44% under blue light and nitrogen starvation conditions. Chlorella sp. AE10 was a good candidate for carbohydrate and lutein productions.

Improving carbohydrate and starch accumulation in Chlorella sp. AE10 by a novel two-stage process with cell dilution.

BACKGROUND: Microalgae are highly efficient cellular factories that capture CO2 and are also alternative feedstock for biofuel production. Carbohydrates, proteins, and lipids are major biochemical components in microalgae. Carbohydrates or starch in microalgae are possible substrates in yeast fermentation for biofuel production. The carbon partitioning in microalgae could be regulated through environmental stresses, such as high concentration of CO2, high light intensity, and nitrogen starvation conditions. It is essential to obtain carbohydrate-rich microalgae via an optimal bioprocess strategy. RESULTS: The carbohydrate accumulation in a CO2 tolerance strain, Chlorella sp. AE10, was investigated with a two-stage process. The CO2 concentration, light intensity, and initial nitrogen concentration were changed drastically in both stages. During the first stage, it was cultivated over 3 days under 1% CO2, a photon flux of 100 µmol m-2 s-1, and 1.5 g L-1 NaNO3. It was cultivated under 10% CO2, 1000 µmol m-2 s-1, and 0.375 g L-1 NaNO3 during the second stage. In addition, two operation modes were compared. At the beginning of the second stage of mode 2, cells were diluted to 0.1 g L-1 and there was no cell dilution in mode 1. The total carbohydrate productivity of mode 2 was increased about 42% compared with that of mode 1. The highest total carbohydrate content and the highest starch content of mode 2 were 77.6% (DW) and 60.3% (DW) at day 5, respectively. The starch productivity was 0.311 g L-1 day-1 and the total carbohydrate productivity was 0.421 g L-1 day-1 in 6 days. CONCLUSIONS: In this study, a novel two-stage process was proposed for improving carbohydrate and starch accumulation in Chlorella sp. AE10. Despite cell dilution at the beginning of the second stage, environmental stress conditions of high concentration of CO2, high light intensity, and limited nitrogen concentration at the second stage were critical for carbohydrate and starch accumulation. Although the cells were diluted, the growths were not inhibited and the carbohydrate productivity was improved. These results were helpful to establish an integrated approach from CO2 capture to biofuel production by microalgae.

Interrogating the Escherichia coli cell cycle by cell dimension perturbations.

Bacteria tightly regulate and coordinate the various events in their cell cycles to duplicate themselves accurately and to control their cell sizes. Growth of Escherichia coli, in particular, follows a relation known as Schaechter's growth law. This law says that the average cell volume scales exponentially with growth rate, with a scaling exponent equal to the time from initiation of a round of DNA replication to the cell division at which the corresponding sister chromosomes segregate. Here, we sought to test the robustness of the growth law to systematic perturbations in cell dimensions achieved by varying the expression levels of mreB and ftsZ We found that decreasing the mreB level resulted in increased cell width, with little change in cell length, whereas decreasing the ftsZ level resulted in increased cell length. Furthermore, the time from replication termination to cell division increased with the perturbed dimension in both cases. Moreover, the growth law remained valid over a range of growth conditions and dimension perturbations. The growth law can be quantitatively interpreted as a consequence of a tight coupling of cell division to replication initiation. Thus, its robustness to perturbations in cell dimensions strongly supports models in which the timing of replication initiation governs that of cell division, and cell volume is the key phenomenological variable governing the timing of replication initiation. These conclusions are discussed in the context of our recently proposed "adder-per-origin" model, in which cells add a constant volume per origin between initiations and divide a constant time after initiation.

Cloning of the A Mating-Type Locus from Lepista nuda and Characterization of Its Genetic Structure.

Lepista nuda (Bull. ex Fr.) Cooke (Tricholomataceae) is an edible fungus with both economic and medical value. Identification of its mating-type loci is important for promoting breeding programs in L. nuda. The A mating-type locus of L. nuda and its flanking region were cloned and characterized in the present study. It contained two homeodomain transcription factor genes (called lna1 and lna2). Lna1 and Lna2 protein harbored conserved motif of homeodomain transcription factor protein. The novel finding of this study was that the gene order around the A locus was mip, lna2, lna1, and ß-fg in L. nuda, which was differed from other edible fungi. In addition, lna1 and lna2 showed divergent, inward transcriptional direction. The phylogenetic tree of HD proteins showed that L. nuda Lna1 and Lna2 were phylogenetically related with Laccaria bicolor. Our results revealed that the A mating-type locus had been subjected to gene rearrangements relative to all other basidiomycetes.

Improving high carbon dioxide tolerance and carbon dioxide fixation capability of Chlorella sp. by adaptive laboratory evolution.

CO2 capture by microalgae is a promising method to reduce greenhouse gas emissions. It is critical to construct a highly efficient way to obtain a microalgal strain tolerant to high CO2 concentrations with high CO2 fixation capability. In this study, two evolved Chlorella sp. strains, AE10 and AE20 were obtained after 31 cycles of adaptive laboratory evolution (ALE) under 10% and 20% CO2, respectively. Both of them grew rapidly in 30% CO2 and the maximal biomass concentration of AE10 was 3.68±0.08g/L, which was 1.22 and 2.94 times to those of AE20 and original strain, respectively. The chlorophyll contents of AE10 and AE20 were significantly higher than those of the original one under 1-30% CO2. The influences of ALE process on biochemical compositions of Chlorella cells were also investigated. This study proved that ALE was an effective approach to improve high CO2 tolerance of Chlorella sp.

Development of SCAR markers to determine the mating types of Lepista nuda protoplast monokaryons.

Lepista nuda (Bull. ex Fr.) Cooke belongs to Tricholomataceae and is an edible fungus with both economic and medical value. Mycelia were isolated from the fruiting bodies of L. nuda and were used to prepare the protoplast monokaryons. One hundred and fifteen monokaryons were obtained and their mating types were determined using somatic incompatibility tests. Protoplast monokaryons segregated into either the A1B1 or the A2B2 mating types. Inter-simple sequence repeats and sequence-related amplified polymorphism fingerprinting were used to analyse the mating types of these protoplast monokaryons and 16 sequence-characterised amplified region primers were developed to efficiently differentiate between the monokaryon mating types. Multiplex PCR analyses were also established. The data presented here outline a method for the precise and rapid identification of protoplast monokaryon mating types, which has the promise to shorten the period required for conventional crossbreeding.