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Nature Communications Aug 2019Brain glioma treatment with checkpoint inhibitor antibodies to cytotoxic T-lymphocyte-associated antigen 4 (a-CTLA-4) and programmed cell death-1 (a-PD-1) was largely...
Brain glioma treatment with checkpoint inhibitor antibodies to cytotoxic T-lymphocyte-associated antigen 4 (a-CTLA-4) and programmed cell death-1 (a-PD-1) was largely unsuccessful due to their inability to cross blood-brain barrier (BBB). Here we describe targeted nanoscale immunoconjugates (NICs) on natural biopolymer scaffold, poly(β-L-malic acid), with covalently attached a-CTLA-4 or a-PD-1 for systemic delivery across the BBB and activation of local brain anti-tumor immune response. NIC treatment of mice bearing intracranial GL261 glioblastoma (GBM) results in an increase of CD8+ T cells, NK cells and macrophages with a decrease of regulatory T cells (Tregs) in the brain tumor area. Survival of GBM-bearing mice treated with NIC combination is significantly longer compared to animals treated with single checkpoint inhibitor-bearing NICs or free a-CTLA-4 and a-PD-1. Our study demonstrates trans-BBB delivery of tumor-targeted polymer-conjugated checkpoint inhibitors as an effective GBM treatment via activation of both systemic and local privileged brain tumor immune response.
Topics: Animals; Antineoplastic Agents, Immunological; Biopolymers; Blood-Brain Barrier; Brain Neoplasms; CTLA-4 Antigen; Cell Line, Tumor; Disease Models, Animal; Female; Glioma; Humans; Immunoconjugates; Malates; Mice; Nanoconjugates; Permeability; Physarum polycephalum; Polymers; Programmed Cell Death 1 Receptor; Treatment Outcome
PubMed: 31462642
DOI: 10.1038/s41467-019-11719-3 -
Plant Disease Jan 2023(Dicks.) Gray, named "Maitake" in Japan, is mainly cultivated in China, Japan and Korea as a rare delicacy (Park et al. 2015). is a medicinal and edible mushroom that...
(Dicks.) Gray, named "Maitake" in Japan, is mainly cultivated in China, Japan and Korea as a rare delicacy (Park et al. 2015). is a medicinal and edible mushroom that can enhance the human immunology system. In recent years, the production of has increased in China due to its high economic value and as a source of livelihood for small scale farmers. From August to September 2017, a serious slime mold disease was observed on G. frondosa under greenhouse conditions in Qingyuan County, Lishui city, Zhejiang Province, China. Incidence was 10 to 30% in most surveyed mushroom greenhouses, sometimes more than 80% in mushroom greenhouses without proper management. The disease reduced production by 10% on average, and over 80% in severe cases. Slime mold disease usually appeared after irrigation, the kelly plasmodia migrate firstly from the root of fruiting body to stem and finally to pileus, then the infected parts became soft and putrid with slime on the surface. Additionally, many other organisms grow on decayed fruiting bodies, such as bacteria, fungi, and insects. The disease can spread rapidly through soil to adjacent fruiting bodies resulting in yield reduction. Samples were collected and cultures were isolated by transferring diseased fruiting bodies with yellow green plasmodia onto 2% water agar medium. Plasmodia were purified through aseptically transferring their edge segment to a new sterile 2% water agar medium, and this procedure was repeated three or four times to free the isolate from contaminating organisms. Purified plasmodia were then placed on the solid bacteriological test medium (SGM), containing glucose, peptone, yeast extract, mineral salts and hematin used in the axenic culture of (Daniel et al. 1964), to verify bacteria presence. Plasmodia were also induced to form sporocarps. Voucher specimens were deposited in the Fungarium of Jiangxi Academy of Agricultural Sciences (FJAAS-M0001) and the Herbarium of the Mycology, Engineering Research Center of Edible and Medicinal Fungi, Chinese Ministry of Education, Jilin Agricultural University (HMJAU-M1561). Sporocarps were stalked, globose to discoid, golden-yellow, 0.9-1.8 mm in height, 0.28-0.55 mm in diameter. Hypothallus was small, thin, orange. Stalks were subulated, about twice to thrice the diameter of the sporotheca, bright orange below, yellow above, furrowed. Peridium was weak, thin, and plated with yellow calcareous flakes. Capillitium was a small meshed, persistent net of tubules with small and yellow angular lime nodes. Spores were globose, free, dark brown to black in mass, purplish brown in transmitted light, 8-10 μm in diameter, smooth under light microscopy. Irregular spinulose spores showed clusters of small warts that are conspicuous under scanning electron microscopy. Plasmodia were yellow green. The 18S ribosomal RNA gene was amplified with primer SMNUR101/NS4 (Rusk et al. 1995; White et al. 1990). The 18S rRNA gene sequence was submitted to GenBank (OP373728) and an 18S rRNA gene phylogenetic tree of Physarum obtained by maximum likelihood analysis (ML) and Bayesian inferences (BI) of 23 taxa and 1,608 aligned positions was produced. Based on sporocarps morphological characteristics, plasmodial cultural traits, and the sequence of 18S rRNA, the slime mold was identified as . A pathogenicity test was performed by gently inoculating a 12 mm diameter circinal patch of SGM with plasmodia on three healthy fruiting bodies of G. frondosa. All treatments were cultured in a mushroom-growing room with temperature 24 to 29 ℃ and relative humidity of 87 to 96%. Three fruiting bodies inoculated with a 12 mm diameter SGM served as controls. All fruiting bodies inoculated with plasmodia showed the same symptom. No symptoms developed on the controls. The pathogen was consistently reisolated from the symptomatic fruiting bodies of and confirmed to be based on cultural, morphological and molecular characteristics, thus fulfilling Kock's postulates. This is the first report of causing yellow rot disease on cultivated . References: Daniel, J. W., et al. 1964. Page 9 in: Methods in Cell Biology. Academic Press, New York. Denchev, C. M. 2008. Mycologia Balc. 5:93. Park, H. S., et al. 2015. Biosci., biotechnol., and biochem. 79:147. Rusk, S. A., et al. 1995. Mycologia. 87:140. White, T. J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA.
PubMed: 36627806
DOI: 10.1094/PDIS-10-22-2419-PDN -
Genome Biology and Evolution May 2020Major phenotypic innovations in social amoeba evolution occurred at the transition between the Polysphondylia and group 4 Dictyostelia, which comprise the model organism...
Major phenotypic innovations in social amoeba evolution occurred at the transition between the Polysphondylia and group 4 Dictyostelia, which comprise the model organism Dictyostelium discoideum, such as the formation of a new structure, the basal disk. Basal disk differentiation and robust stalk formation require the morphogen DIF-1, synthesized by the polyketide synthase StlB, the des-methyl-DIF-1 methyltransferase DmtA, and the chlorinase ChlA, which are conserved throughout Dictyostelia. To understand how the basal disk and other innovations evolved in group 4, we sequenced and annotated the Polysphondylium violaceum (Pvio) genome, performed cell type-specific transcriptomics to identify cell-type marker genes, and developed transformation and gene knock-out procedures for Pvio. We used the novel methods to delete the Pvio stlB gene. The Pvio stlB- mutants formed misshapen curly sorogens with thick and irregular stalks. As fruiting body formation continued, the upper stalks became more regular, but structures contained 40% less spores. The stlB- sorogens overexpressed a stalk gene and underexpressed a (pre)spore gene. Normal fruiting body formation and sporulation were restored in Pvio stlB- by including DIF-1 in the supporting agar. These data indicate that, although conserved, stlB and its product(s) acquired both a novel role in the group 4 Dictyostelia and a role opposite to that in its sister group.
Topics: Genome, Protozoan; Myxomycetes; Polyketide Synthases; Protozoan Proteins
PubMed: 32386295
DOI: 10.1093/gbe/evaa079 -
Applications in Plant Sciences 2024Traditional moist chamber cultures (MCs) prepared in aseptic laboratory environments using sterile Petri dishes are commonly used to quantify the microbiota of...
PREMISE
Traditional moist chamber cultures (MCs) prepared in aseptic laboratory environments using sterile Petri dishes are commonly used to quantify the microbiota of rough-bark tree species and woody vines. MCs are typically expensive and may be difficult to make, so a less expensive option made from easily available supplies was developed. These cost-friendly MCs were compared with standard laboratory methods to demonstrate their efficacy.
METHODS AND RESULTS
Modified MCs were made using inexpensive, store-bought supplies; compared to a standard laboratory setting, the modified MCs are shown to be less expensive with a faster setup time and larger size that facilitates a variety of tree and woody vine species. MC use resulted in the discovery of new species of fungi and myxomycetes with associated locality records. We provide detailed instructions for creating modified MCs, as well as a list of myxomycete species and their associated bark characteristics, pH values, and water-holding capacity.
CONCLUSIONS
This new, low-cost MC technique makes the study of microbiota more inclusive and accessible for those in research laboratories, classrooms, and homes, including both amateurs and professionals. MCs are easy to prepare, versatile, and applicable for many areas of botany and the biological sciences, potentially allowing exploration into unexplored areas in urban ecosystems.
PubMed: 38638615
DOI: 10.1002/aps3.11578 -
Biodiversity Data Journal 2024As a result of the ten years (2012-2022) work under the critical revision of the genera of Reticulariaceae, a set of papers was published. Collection data of hundreds of...
BACKGROUND
As a result of the ten years (2012-2022) work under the critical revision of the genera of Reticulariaceae, a set of papers was published. Collection data of hundreds of specimens, used as a material for these studies, were provided as supplements of corresponding papers, but remained unpublished in biodiversity databases.
NEW INFORMATION
Here, we represent an occurrence dataset "Barcoded Reticulariaceae of the World", published in GBIF. It includes data on 523 myxomycete collections (including 36 types) gathered from five continents and spanning 24 countries. The dataset encompasses 43 distinct species and one subspecies of myxomycetes, including rare, endemic, and recently-described taxa. Species included to the database mainly belong to the genera , , , , and (Reticulariaceae), but as well and (Cribrariaceae). Nearly all of the research material, with the exception of several old collections, underwent molecular barcoding, primarily involving the 18S rDNA gene, but also the elongation factor 1α gene and mitochondrial cytochrome oxidase subunit I gene. For those sequences that are stored in the NCBI GenBank, accession numbers are provided in the dataset. Newly-described species make up a significant part of the studied herbarium collections; many of them can be characterised as common for their region. A particularly high level of taxonomic novelty is observed in Australia, which may be explained by the endemism of the local myxomycete biota.
PubMed: 38469226
DOI: 10.3897/BDJ.12.e115630 -
Philosophical Transactions of the Royal... Mar 2021The slime mould , an aneural organism, uses information from previous experiences to adjust its behaviour, but the mechanisms by which this is accomplished remain...
The slime mould , an aneural organism, uses information from previous experiences to adjust its behaviour, but the mechanisms by which this is accomplished remain unknown. This article examines the possible role of oscillations in learning and memory in slime moulds. Slime moulds share surprising similarities with the network of synaptic connections in animal brains. First, their topology derives from a network of interconnected, vein-like tubes in which signalling molecules are transported. Second, network motility, which generates slime mould behaviour, is driven by distinct oscillations that organize into spatio-temporal wave patterns. Likewise, neural activity in the brain is organized in a variety of oscillations characterized by different frequencies. Interestingly, the oscillating networks of slime moulds are not precursors of nervous systems but, rather, an alternative architecture. Here, we argue that comparable information-processing operations can be realized on different architectures sharing similar oscillatory properties. After describing learning abilities and oscillatory activities of , we explore the relation between network oscillations and learning, and evaluate the organism's global architecture with respect to information-processing potential. We hypothesize that, as in the brain, modulation of spontaneous oscillations may sustain learning in slime mould. This article is part of the theme issue 'Basal cognition: conceptual tools and the view from the single cell'.
Topics: Adaptation, Biological; Learning; Memory; Physarum polycephalum; Signal Transduction
PubMed: 33487112
DOI: 10.1098/rstb.2019.0757 -
Scientific Reports Feb 2022The maternal/uniparental inheritance of mitochondria is controlled by the selective elimination of paternal/uniparental mitochondria and digestion of their mitochondrial...
The maternal/uniparental inheritance of mitochondria is controlled by the selective elimination of paternal/uniparental mitochondria and digestion of their mitochondrial DNA (mtDNA). In isogamy, the selective digestion of mtDNA in uniparental mitochondria is initiated after mating and is completed prior to the elimination of mitochondria, but the molecular mechanism of the digestion of uniparental mtDNA remains unknown. In this study, we developed a semi-in vitro assay for DNase, wherein the digestion of mitochondrial nucleoids (mt-nucleoids) was microscopically observed using isolated mitochondria from Physarum polycephalum and the DNase involved in uniparental inheritance was characterized. When myxamoebae of AI35 and DP246 are crossed, mtDNA and mt-nucleoid from only the DP246 parent are digested. The digestion of mt-nucleoids was observed in zygotes 3 h after plating for mating. During the digestion of mt-nucleoids, mitochondrial membrane integrity was maintained. In the semi-in vitro assay, the digestion of mt-nucleoids was only observed in the presence of Mg at pH 7.5-9.0. Moreover, such Mg-dependent DNase activity was specifically detected in mitochondria isolated from zygotes 3 h after plating for mating. Therefore, Mg-dependent DNase is potentially involved in uniparental inheritance. Our findings provide insights into the DNase involved in uniparental inheritance and its regulatory mechanism.
Topics: DNA, Mitochondrial; Deoxyribonucleases; Hydrogen-Ion Concentration; Magnesium; Mitochondria; Mitochondrial Membranes; Physarum polycephalum; Zygote
PubMed: 35194142
DOI: 10.1038/s41598-022-06920-2 -
Bio Systems Oct 2021This research addresses the interactions between the unicellular slime mold Physarum polycephalum and a red yeast in a spatial ecosystem over week-long imaging...
This research addresses the interactions between the unicellular slime mold Physarum polycephalum and a red yeast in a spatial ecosystem over week-long imaging experiments. An inverse relationship between the growth rates of both species is shown, where P. polycephalum has positive growth when the red yeast has a negative growth rate and vice versa. The data also captures successional and oscillatory dynamics between both species. An advanced image analysis methodology for semantic segmentation is used to quantify population density over time, for all components of the ecosystem. We suggest that P. polycephalum is capable of exhibiting a sustainable feeding strategy by depositing a nutritive slime trail, allowing yeast to serve as a periodic food source. This opens a new direction of P. polycephalum research, where the population dynamics of spatial ecosystems can be readily quantified and complex ecological dynamics can be studied.
Topics: Deep Learning; Ecosystem; Microbiological Phenomena; Physarum polycephalum; Population Dynamics
PubMed: 34271083
DOI: 10.1016/j.biosystems.2021.104483 -
Biology Letters Feb 2023In metazoans, the expression of key phenotypic traits is sensitive to two- and three-way interactions between variation in mitochondrial DNA, nuclear DNA and the...
In metazoans, the expression of key phenotypic traits is sensitive to two- and three-way interactions between variation in mitochondrial DNA, nuclear DNA and the external environment. Whether gene--environment interactions affect phenotypes in single-celled eukaryotes is poorly studied, except in a few species of yeast and fungi. We developed a genetic panel of the unicellular slime mould, containing strains differing in mitochondrial and nuclear DNA haplotypes. The panel also included two strains harbouring a selfishly replicating mitochondrial-fusion (mF) plasmid that could affect phenotype expression. We assayed movement and growth rate differences among the strains across two temperature regimes: 24° and 28°C. We found that the slime mould's growth rate, but not movement, is affected by G × G × E interactions. Predictably, mtDNA × nDNA interactions significantly affected both traits. The inter-trait correlation across the strains in each temperature regime was positive. Surprisingly, the mF plasmid had no negative effects on our chosen traits. Our study is the first to demonstrate genetic regulation of phenotype expression in a unicellular slime mould. The genetic effect on phenotypes manifests via epistatic interactions with the thermal environment, thus shedding new light on the role of G × G × E interactions in trait evolution in protists.
Topics: Physarum polycephalum; DNA, Mitochondrial; Mitochondria; Plasmids; Phenotype
PubMed: 36789533
DOI: 10.1098/rsbl.2022.0494 -
Mycoscience 2022A new species of (), described herein as , was recovered in the field on ground litter from mountain subtropical forests (Phia Oắc - Phia Đén National Park) of...
: a new species of myxomycetes with reticulate spores from Phia Oắc - Phia Đén National Park (northern Vietnam) supported by molecular phylogeny and morphological analysis.
A new species of (), described herein as , was recovered in the field on ground litter from mountain subtropical forests (Phia Oắc - Phia Đén National Park) of northern Vietnam. Morphological details were examined by light and scanning electron microscopy. The species is characterized by a distinct and unique combination of morphological features, including a bright blue, shiny and very thin membranous peridium, a small dome-shaped columella, rigid, straight, branched, brown capillitial threads which gradually become pale at the periphery and finally colorless at the tips and small-meshed, banded-reticulate spores with 9-12 meshes across the spore diameter and solid walls without perforations 0.3-0.5 µm high. The stability of the taxonomic characters of is supported by two well-developed collections found in 2018 and 2019. Partial sequences of three molecular markers (SSU, EF1α, COI) for both collections are identical. A two-gene phylogeny of the first two markers displays the two known accessions as a well-separated entity and indicates affinity of the new species with (the type taxon of the genus), , and several nivicolous species.
PubMed: 37090470
DOI: 10.47371/mycosci.2022.05.003