G3 (Bethesda). 2025 Feb 18:jkaf033. doi: 10.1093/g3journal/jkaf033. Online ahead of print.

ABSTRACT

Facultatively symbiotic corals provide important experimental models to explore the establishment, maintenance, and breakdown of the mutualism between corals and members of the algal family Symbiodiniaceae. Here, we report the de novo chromosome-scale genome assembly and annotation of the facultatively symbiotic, temperate coral Astrangia poculata. Though widespread segmental/tandem duplications of genomic regions were detected, we did not find strong evidence of a whole genome duplication (WGD) event. Comparison of the gene arrangement between A. poculata and the tropical coral Acropora millepora revealed considerable conserved colinearity despite ∼415 million years of divergence. Gene families related to sperm hyperactivation and innate immunity, including lectins, were found to contain more genes in A. millepora relative to A. poculata. Sperm hyperactivation in A. millepora is expected given the extreme requirements of gamete competition during mass spawning events in tropical corals, while lectins are important in the establishment of coral-algal symbiosis. By contrast, gene families involved in sleep promotion, feeding suppression, and circadian sleep/wake cycle processes were expanded in A. poculata. These expanded gene families may play a role in A. poculata's ability to enter a dormancy-like state ("winter quiescence") to survive freezing temperatures at the northern edges of the species' range.

PMID:39964876 | DOI:10.1093/g3journal/jkaf033

Elife. 2025 Feb 18;13:RP97682. doi: 10.7554/eLife.97682.

ABSTRACT

Untranslated regions (UTRs) contain crucial regulatory elements for RNA stability, translation and localization, so their integrity is indispensable for gene expression. Approximately 3.7% of genetic variants associated with diseases occur in UTRs, yet a comprehensive understanding of UTR variant functions remains limited due to inefficient experimental and computational assessment methods. To systematically evaluate the effects of UTR variants on RNA stability, we established a massively parallel reporter assay on 6555 UTR variants reported in human disease databases. We examined the RNA degradation patterns mediated by the UTR library in two cell lines, and then applied LASSO regression to model the influential regulators of RNA stability. We found that UA dinucleotides and UA-rich motifs are the most prominent destabilizing element. Gain of UA dinucleotide outlined mutant UTRs with reduced stability. Studies on endogenous transcripts indicate that high UA-dinucleotide ratios in UTRs promote RNA degradation. Conversely, elevated GC content and protein binding on UA dinucleotides protect high-UA RNA from degradation. Further analysis reveals polarized roles of UA-dinucleotide-binding proteins in RNA protection and degradation. Furthermore, the UA-dinucleotide ratio of both UTRs is a common characteristic of genes in innate immune response pathways, implying a coordinated stability regulation through UTRs at the transcriptomic level. We also demonstrate that stability-altering UTRs are associated with changes in biobank-based health indices, underscoring the importance of precise UTR regulation for wellness. Our study highlights the importance of RNA stability regulation through UTR primary sequences, paving the way for further exploration of their implications in gene networks and precision medicine.

PMID:39964837 | DOI:10.7554/eLife.97682

Proc Natl Acad Sci U S A. 2025 Feb 25;122(8):e2420961122. doi: 10.1073/pnas.2420961122. Epub 2025 Feb 18.

ABSTRACT

Protein Serine Kinase H1 (PSKH1) was recently identified as a crucial factor in kidney development and is overexpressed in prostate, lung, and kidney cancers. However, little is known about PSKH1 regulatory mechanisms, leading to its classification as a "dark" kinase. Here, we used biochemistry and mass spectrometry to define PSKH1's consensus substrate motif, protein interactors, and how interactors, including Ca2+ sensor proteins, promote or suppress activity. Intriguingly, despite the absence of a canonical Calmodulin binding motif, Ca2+-Calmodulin activated PSKH1 while, in contrast, the ER-resident Ca2+ sensor of the Cab45, Reticulocalbin, Erc55, Calumenin (CREC) family, Reticulocalbin-3, suppressed PSKH1 catalytic activity. In addition to antagonistic regulation of the PSKH1 kinase domain by Ca2+ sensing proteins, we identified UNC119B as a protein interactor that activates PSKH1 via direct engagement of the kinase domain. Our findings identify complementary allosteric mechanisms by which regulatory proteins tune PSKH1's catalytic activity and raise the possibility that different Ca2+ sensors may act more broadly to tune kinase activities by detecting and decoding extremes of intracellular Ca2+ concentrations.

PMID:39964718 | DOI:10.1073/pnas.2420961122

Eur J Nutr. 2025 Feb 18;64(2):97. doi: 10.1007/s00394-025-03600-5.

ABSTRACT

PURPOSE: Recent studies indicate that shift work may affect workers' eating habits and overall well-being. This study aimed to assess differences in eating patterns, sleep quality, and mental health between Italian shift and non-shift workers, with a focus on individual chronotype and the type of shift work (day vs. night shift).

METHODS: The cross-sectional study involved 322 subjects (166 shift and 156 non-shift workers). Eating habits were evaluated using a 7-day diary and the Medi-Lite questionnaire. Sleep quality was assessed with the Pittsburgh Sleep Quality Index (PSQI), and mental health with the Depression Anxiety Stress Scales (DASS). Individual chronotype was defined using the Morningness-Eveningness Questionnaire.

RESULTS: No significant differences in daily energy, macronutrient, and micronutrient intake between the two groups, nor in the temporal pattern of eating. However, shift workers had significantly (p < 0.05) lower adherence to the Mediterranean diet (MD) (7.6 ± 2.3 vs 8.1 ± 2.2) compared to non-shift workers. Shift workers also reported significantly poorer sleep quality (mean PSQI score 7.6 ± 3.7 vs. 5.8 ± 3.0) and higher levels of anxiety and stress symptoms. Among shift workers, those with an evening chronotype had significantly lower MD adherence than those with a morning chronotypes. Additionally, night shift workers experienced more sleep disturbances compared to day ones.

CONCLUSION: Shift workers reported lower MD adherence, poorer sleep quality, and a higher prevalence of anxiety and stress symptoms compared to a similar group of non-shift workers. Evening chronotypes and night shift work were associated with worse eating habits and sleep quality.

PMID:39964501 | DOI:10.1007/s00394-025-03600-5

Genomics Proteomics Bioinformatics. 2025 Feb 18:qzaf009. doi: 10.1093/gpbjnl/qzaf009. Online ahead of print.

ABSTRACT

In recent years, computational methods for quantifying cell type proportions from transcription data have gained significant attention, particularly those reference-based methods which have demonstrated high accuracy. However, there is currently a lack of comprehensive evaluation and guidance for available reference-based deconvolution methods in cell proportion deconvolution analysis. In this study, we introduce Deconvolution Evaluator (Deconer), a comprehensive toolkit for the evaluation of reference-based deconvolution methods. Deconer provides various simulated and real gene expression datasets, including both bulk and single-cell sequencing data, and offers multiple visualization interfaces. By utilizing Deconer, we conducted systematic comparisons of 16 reference-based deconvolution methods from different perspectives, including method robustness, accuracy in deconvolving rare components, signature gene selection, and building external reference. We also performed an in-depth analysis of the application scenarios and challenges in cell proportion deconvolution methods. Finally, we provided constructive suggestions for users in selecting and developing cell proportion deconvolution algorithms. This work presents novel insights to researchers, assisting them in choosing appropriate toolkits, applying solutions in clinical contexts, and advancing the development of deconvolution tools tailored to gene expression data. The tutorials, manual, source code, and demo data of Deconer are publicly available at https://honchkrow.github.io/Deconer/.

PMID:39963994 | DOI:10.1093/gpbjnl/qzaf009

Mol Biol Evol. 2025 Feb 18:msaf045. doi: 10.1093/molbev/msaf045. Online ahead of print.

ABSTRACT

In the wake of pandemics like COVID-19, which have zoonotic origins, the role of wildlife as reservoirs for emerging infectious diseases has garnered heightened attention. Migratory birds, traversing continents, represent a potent but under-researched vector for the spread of infectious diseases, including novel coronaviruses. This study delves into the genetic diversity and transmission dynamics of coronaviruses in migratory birds, presenting pivotal findings. From April 2019 to April 2023, we screened 5,263 migratory bird samples collected from Shanghai, China, identifying 372 coronavirus-positive samples belonging to five avian-related coronavirus subgenera and subsequently obtaining 120 complete genome sequences. To facilitate further research with a global perspective, the study curated all available 19,000 avian-associated CoVs and expanded the original 12 species to 16, including three novel coronavirus species identified in our study and one re-classified species from the public domain. The study illuminates the intricate genetic evolution and transmission dynamics of birds-related coronaviruses on a global scale. A notable aspect of our research is the identification of complex recombination patterns within the spike protein across different virus species and subgenera, highlighting migratory birds as a reservoir of coronavirus. Notably, the coronaviruses found in migratory birds, predominantly from the orders Anseriformes, Charadriiformes, and Pelecaniformes, with domestic ducks from Anseriformes playing a key role in bridging the transmission of coronaviruses between migratory and non-migratory birds. These findings reveal the genetic and recombination characteristics of coronaviruses in migratory birds, emphasizing the critical role of ecologically pivotal bird species in coronavirus transmission and genetic diversity shaping.

PMID:39963938 | DOI:10.1093/molbev/msaf045

Front Plant Sci. 2025 Feb 3;16:1559814. doi: 10.3389/fpls.2025.1559814. eCollection 2025.

NO ABSTRACT

PMID:39963532 | PMC:PMC11830810 | DOI:10.3389/fpls.2025.1559814

Comput Struct Biotechnol J. 2024 Sep 27;23:3575-3583. doi: 10.1016/j.csbj.2024.09.018. eCollection 2024 Dec.

ABSTRACT

Accessible and easy-to-use standardized bioinformatics workflows are necessary to advance microbiome research from observational studies to large-scale, data-driven approaches. Standardized multi-omics data enables comparative studies, data reuse, and applications of machine learning to model biological processes. To advance broad accessibility of standardized multi-omics bioinformatics workflows, the National Microbiome Data Collaborative (NMDC) has developed the Empowering the Development of Genomics Expertise (NMDC EDGE) resource, a user-friendly, open-source web application (https://nmdc-edge.org). Here, we describe the design and main functionality of the NMDC EDGE resource for processing metagenome, metatranscriptome, natural organic matter, and metaproteome data. The architecture relies on three main layers (web application, orchestration, and execution) to ensure flexibility and expansion to future workflows. The orchestration and execution layers leverage best practices in software containers and accommodate high-performance computing and cloud computing services. Further, we have adopted a robust user research process to collect feedback for continuous improvement of the resource. NMDC EDGE provides an accessible interface for researchers to process multi-omics microbiome data using production-quality workflows to facilitate improved data standardization and interoperability.

PMID:39963423 | PMC:PMC11832004 | DOI:10.1016/j.csbj.2024.09.018

Small Methods. 2025 Feb 17:e2402111. doi: 10.1002/smtd.202402111. Online ahead of print.

ABSTRACT

Spatially resolved transcriptomics (SRT) has emerged as a transformative technology for elucidating cellular organization and tissue architecture. However, a significant challenge remains in identifying pathology-relevant spatial functional landscapes within the tissue microenvironment, primarily due to the limited integration of cell-cell communication dynamics. To address this limitation, SpaDCN, a Spatially Dynamic graph Convolutional Network framework is proposed, which aligns cell-cell communications and gene expression within a spatial context to reveal the spatial functional regions with the coherent cellular organization. To effectively transfer the influence of cell-cell communications on expression variation, SpaDCN respectively generates the node layer and edge layer of spatial graph representation from expression data and the ligand-receptor complex contributions and then employs a dynamic graph convolution to switch the propagation of node graph and edge graph. It is demonstrated that SpaDCN outperforms existing methods in identifying spatial domains and denoising expression across various platforms and species. Notably, SpaDCN excels in identifying marker genes with significant prognostic potential in cancer tissues. In conclusion, SpaDCN offers a powerful and precise tool for spatial domain detection in spatial transcriptomics, with broad applicability across various tissue types and research disciplines.

PMID:39962819 | DOI:10.1002/smtd.202402111

Mol Syst Biol. 2025 Feb 17. doi: 10.1038/s44320-025-00090-9. Online ahead of print.

ABSTRACT

Algorithms that constrain metabolic network models with enzyme levels to predict metabolic activity assume that changes in enzyme levels are indicative of flux variations. However, metabolic flux can also be regulated by other mechanisms such as allostery and mass action. To systematically explore the relationship between fluctuations in enzyme expression and flux, we combine available yeast proteomic and fluxomic data to reveal that flux changes can be best predicted from changes in enzyme levels of pathways, rather than the whole network or only cognate reactions. We implement this principle in an 'enhanced flux potential analysis' (eFPA) algorithm that integrates enzyme expression data with metabolic network architecture to predict relative flux levels of reactions including those regulated by other mechanisms. Applied to human data, eFPA consistently predicts tissue metabolic function using either proteomic or transcriptomic data. Additionally, eFPA efficiently handles data sparsity and noisiness, generating robust flux predictions with single-cell gene expression data. Our approach outperforms alternatives by striking an optimal balance, evaluating enzyme expression at pathway level, rather than either single-reaction or whole-network levels.

PMID:39962320 | DOI:10.1038/s44320-025-00090-9

Nat Genet. 2025 Feb 17. doi: 10.1038/s41588-025-02098-1. Online ahead of print.

NO ABSTRACT

PMID:39962239 | DOI:10.1038/s41588-025-02098-1

Sci Rep. 2025 Feb 17;15(1):5737. doi: 10.1038/s41598-025-88417-2.

ABSTRACT

Triple-negative breast cancer (TNBC) is difficult to treat and has a low five-year survival rate. In South Africa, a large percentage of the population still relies on traditional plant-based medicine. To establish the utility of both methanol and water-soluble extracts from the leaves of Tulbaghia violacea, cytotoxicity assays were carried out to establish the IC50 values against a TNBC cell line. Cell cycle and apoptosis assays were carried out using the extracts. To identify the molecular compounds, present in water-soluble leaf extracts, NMR spectroscopy was performed. Compounds of interest were then used in computational docking studies with the anti-apoptotic protein COX-2. The IC50 values for the water- and methanol-soluble extracts were determined to be 400 and 820 µg/mL, respectively. The water-soluble extract induced apoptosis in the TNBC cell line to a greater extent than in the normal cell line. RNAseq indicated that there was an increase in the transcription of pro-apoptotic genes in the TNBC cell line. The crude extract also caused these cells to stall in the S phase. Of the 61 compounds identified in this extract, five demonstrated a high binding affinity for COX-2. Based on these findings, the compounds within the extract show significant potential for further investigation as candidates for the development of cancer therapeutics, particularly for TNBC.

PMID:39962120 | DOI:10.1038/s41598-025-88417-2

J Nutr Biochem. 2025 Feb 15:109869. doi: 10.1016/j.jnutbio.2025.109869. Online ahead of print.

ABSTRACT

Coffee brew is an integral part of the individual diet worldwide. Roasted coffee contains numerous bioactive substances whose significance for health is investigated in nutritional studies. Food biomarkers are recommended to correlate coffee consumption and health effects in the most unbiased way possible. Metabolites of atractyligenin derivatives from roasted coffee have been suggested as candidate analytes indicating coffee consumption. UHPLC-MS/MS analysis revealed that atractyligenin (1), 2-O-β-D-glucosylatractyligenin and 3'-O-β-D-glucosyl-2'-O-isovaleryl-2-O-β-D-glucosylatractyligenin were extracted into coffee brew. Their concentrations in filtered and unfiltered coffee did not differ significantly, suggesting independence from the preparation method. In a coffee intervention study (n=12, female/male 6/6), atractyligenin metabolites were not detectable in plasma after three days of coffee abstinence. After coffee, atractyligenin (1) and atractyligenin-19-O-D-glucuronide (M1) were the quantitatively dominant atractyligenin metabolites in plasma and showed two peaks each after 0.5 and 10 h, respectively. Half-lives after the first cmax in plasma were ∼0.31 h. 1 and M1 were detectable in plasma, indicating coffee consumption for up to 24 h after one serving. Within 10 h, ∼13.7% of the atractyligenin glycosides supplied by coffee brew were excreted in urine as metabolites 1 and M1. Metabolites 2β-hydroxy-15-oxoatractylan-4α-carboxy-19-O-β-d-glucuronide (M2) and 2β-hydroxy-15-oxoatractylan-4α-carboxylic acid-2-O-β-d-glucuronide (M3) were detected in only some samples and appeared unreliable as indicators for coffee consumption. No concentration differences between female and male study participants were observed in plasma and urine. In conclusion atractyligenin and its 19-O-β-D-glucuronide are promising markers of Arabica coffee consumption in plasma and urine for both men and women, independent of the brewing method.

PMID:39961551 | DOI:10.1016/j.jnutbio.2025.109869

Funct Plant Biol. 2025 Feb;52:FP24051. doi: 10.1071/FP24051.

ABSTRACT

We applied a systems biology approach to gain a deep insight into the regulatory mechanisms of barley (Hordeum vulgare ) under drought and waterlogging stress conditions. To identify informative models related to stress conditions, we constructed meta-analysis and two distinct weighted gene co-expression networks. We then performed module trait association analyses. Additionally, we conducted functional enrichment analysis of significant modules to shed light on the biological performance of underlying genes in the two contrasting stresses. In the next step, we inferred the gene regulatory networks between top hub genes of significant modules, kinases, and transcription factors (TFs) using a machine learning algorithm. Our results showed that at power=10, the scale-free topology fitting index (R2) was higher than 0.8 and the connectivity mean became stable. We identified 31 co-expressed gene modules in barley, with 13 and 14 modules demonstrating significant associations with drought and waterlogging stress, respectively. Functional enrichment analysis indicated that these stress-responsive modules are involved in critical processes, including ADP-rybosylation factors (ARF) protein signal transduction, ethylene-induced autophagy, and phosphoric ester hydrolase activity. Specific TFs and kinases, such as C2C2-GATA, HB-BELL, and MADS-MIKC, were identified as key regulators under these stress conditions. Furthermore, certain TFs and kinases established unique connections with hub genes in response to waterlogging and drought conditions. These findings enhance our understanding of the molecular networks that modulate barley's response to drought and waterlogging stresses, offering insights into the regulatory mechanisms essential for stress adaptation.

PMID:39960829 | DOI:10.1071/FP24051

Elife. 2025 Feb 17;12:RP87930. doi: 10.7554/eLife.87930.

ABSTRACT

Cell mechanical properties determine many physiological functions, such as cell fate specification, migration, or circulation through vasculature. Identifying factors that govern the mechanical properties is therefore a subject of great interest. Here, we present a mechanomics approach for establishing links between single-cell mechanical phenotype changes and the genes involved in driving them. We combine mechanical characterization of cells across a variety of mouse and human systems with machine learning-based discriminative network analysis of associated transcriptomic profiles to infer a conserved network module of five genes with putative roles in cell mechanics regulation. We validate in silico that the identified gene markers are universal, trustworthy, and specific to the mechanical phenotype across the studied mouse and human systems, and demonstrate experimentally that a selected target, CAV1, changes the mechanical phenotype of cells accordingly when silenced or overexpressed. Our data-driven approach paves the way toward engineering cell mechanical properties on demand to explore their impact on physiological and pathological cell functions.

PMID:39960760 | DOI:10.7554/eLife.87930

J Occup Health. 2025 Feb 17:uiaf013. doi: 10.1093/joccuh/uiaf013. Online ahead of print.

ABSTRACT

The novel properties and functions of nanomaterials have naturally alerted the toxicologists to the fact that such materials may also have novel effects on the human body and living organisms. In particular, materials with high stability or biopersisteny have been shown to have a tendency to accumulate in the body, leading to chronic toxicity including carcinogenicity. However, at the early stages of toxicity research, the information is often limited to the effects of short-term exposure studies, and findings on chronic effects are very much delayed. In this context, it was rather exceptional that studies on multiwall carbon nanotubes (MWCNTs) have started with the verification of their potential to induce mesothelioma. This toxicological endpoint was expected on the basis of existing knowledge of asbestos and asbestos-like fiber particles. This movement has led to the achievement of the original mission of the "Modern Toxicology", which is "to achieve a win-win situation where both industrial promotion and safety assurance are ensured by communicating and sharing toxicity information to developers and consumers at a stage before mass production and consumption begins, that is, before massive exposure of the general public begins". Inaccurate toxicity assessments of asbestos in the 1980s and 1990s allowed its spread to our living environment, which is difficult to decontaminate, and the damage still continues to this day. However, the case described here could be an example of realizing the proposition that 'nanomaterials, the flagship of high technology, must not repeat the same mistakes.'

PMID:39960454 | DOI:10.1093/joccuh/uiaf013

Plant Cell Environ. 2025 Feb 17. doi: 10.1111/pce.15430. Online ahead of print.

ABSTRACT

Legumes have the beneficial capacity to establish symbiotic interactions with rhizobia, which provide their host plants with fixed nitrogen. However, in the presence of nitrogen, this process is rapidly repressed to avoid unnecessary investments of carbon in the symbiosis. Several players involved in regulating nodulation in response to nitrate availability have been identified, including peptide hormones, microRNAs and transcription factors. Nevertheless, how these molecular players are linked to each other and what underlying molecular mechanisms are at play to inhibit nodulation remain unresolved. Nitrate-mediated control of nodulation seems to differ between model legumes, such as Medicago and Lotus, compared to legume crops such as soybean. A deeper understanding of these regulatory processes, particularly in soybean, is expected to contribute to establishing increased nodulation efficiency in modern agricultural systems, hence improving sustainability by reducing the need for environmentally hazardous nitrogen fertilizers. This review describes the state of the art of nitrate-regulated nodulation in soybean, while drawing parallels with molecular mechanisms described in other legumes and addressing knowledge gaps that require future study.

PMID:39960031 | DOI:10.1111/pce.15430

Annu Rev Cancer Biol. 2024 Jun;8(1):15-33. doi: 10.1146/annurev-cancerbio-062722-013740. Epub 2023 Dec 6.

ABSTRACT

The non-receptor tyrosine phosphatase SHP2 has been at the center of cell signaling research for three decades. SHP2 is required to fully activate the RTK-RAS-ERK cascade, although the underlying mechanisms are not completely understood. PTPN11, coding for SHP2, is the first identified proto-oncogene that encodes a tyrosine phosphatase, with dominantly activating mutations detected in leukemias and solid tumors. However, SHP2 has been shown to have pro- and anti-oncogenic effects, and the most recent data reveal opposite activities of SHP2 in tumor cells and microenvironment cells. Allosteric SHP2 inhibitors show promising anti-tumor effects and overcome resistance to inhibitors of RAS-ERK signaling in animal models. Many clinical trials with orally bioactive SHP2 inhibitors, alone or combined with other regimens, are ongoing for a variety of cancers worldwide, with therapeutic outcomes yet unknown. This review discusses the multi-faceted SHP2 functions in oncogenesis, preclinical studies and clinical trials with SHP2 inhibitors in oncological treatment.

PMID:39959686 | PMC:PMC11824402 | DOI:10.1146/annurev-cancerbio-062722-013740

Liver Res. 2024 Dec 5;8(4):218-236. doi: 10.1016/j.livres.2024.12.001. eCollection 2024 Dec.

ABSTRACT

Liver cancer ranks as the sixth most common cancer globally, with hepatocellular carcinoma (HCC) accounting for approximately 75%-85% of cases. Most patients present with moderately advanced disease, while those with advanced HCC face limited and ineffective treatment options. Despite diagnostic efforts, no ideal tumor marker exists to date, highlighting the urgent clinical need for improved early detection of HCC. A key research objective is the development of assays that target specific pathways involved in HCC progression. This review explores the pathological origin and development of HCC, providing insights into the mechanistic rationale, clinical statistics, and the advantages and limitations of commonly used diagnostic tumor markers. Additionally, it discusses the potential of emerging biomarkers for early diagnosis and offers a brief overview of relevant assay methodologies. This review aims to summarize existing markers and investigate new ones, providing a basis for subsequent research.

PMID:39958920 | PMC:PMC11771281 | DOI:10.1016/j.livres.2024.12.001

MicroPubl Biol. 2025 Jan 31;2025. doi: 10.17912/micropub.biology.001485. eCollection 2025.

ABSTRACT

DNA fiber assays are indispensable tools for studying DNA damage and replication stress responses in vivo at the single replication fork level. These assays typically rely on antibodies recognizing IdU and CldU. Historically, the availability of CldU-reactive antibodies has been limited to one reagent (clone BU1/75(ICR1)). We validated two alternative antibodies for CldU detection in DNA fiber assays. One of these antibodies can be readily paired with a common IdU-reactive antibody, and we confirmed that it produces quantitatively similar CldU track length results vis-à-vis the BU1/75 antibody. The new reagents should boost versatility of DNA fiber assays, facilitating DNA replication research.

PMID:39958910 | PMC:PMC11829213 | DOI:10.17912/micropub.biology.001485