Primed State Research Articles

On State Ideals and State Relative Annihilators in De Morgan State Residuated Lattices

The concept of state has been considered in commutative and noncommutative logical systems, and their properties are at the center for the development of an algebraic investigation of probabilistic models for those algebras. This article mainly focuses on the study of the lattice of state ideals in De Morgan state residuated lattices (DMSRLs). First, we prove that the lattice of all state ideals S ℐ L of a DMSRL L , τ is a coherent frame. Then, we characterize the DMSRL for which the lattice S ℐ L is a Boolean algebra. In addition, we bring in the concept of state relative annihilator of a given nonempty subset with respect to a state ideal in DMSRL and investigate various properties. We prove that state relative annihilators are a particular kind of state ideals. Finally, we investigate the notion of prime state ideal in DMSRL and establish the prime state ideal theorem.

All-atom molecular dynamics simulations of Synaptotagmin-SNARE-complexin complexes bridging a vesicle and a flat lipid bilayer.

Synaptic vesicles are primed into a state that is ready for fast neurotransmitter release upon Ca2+-binding to Synaptotagmin-1. This state likely includes trans-SNARE complexes between the vesicle and plasma membranes that are bound to Synaptotagmin-1 and complexins. However, the nature of this state and the steps leading to membrane fusion are unclear, in part because of the difficulty of studying this dynamic process experimentally. To shed light into these questions, we performed all-atom molecular dynamics simulations of systems containing trans-SNARE complexes between two flat bilayers or a vesicle and a flat bilayer with or without fragments of Synaptotagmin-1 and/or complexin-1. Our results need to be interpreted with caution because of the limited simulation times and the absence of key components, but suggest mechanistic features that may control release and help visualize potential states of the primed Synaptotagmin-1-SNARE-complexin-1 complex. The simulations suggest that SNAREs alone induce formation of extended membrane-membrane contact interfaces that may fuse slowly, and that the primed state contains macromolecular assemblies of trans-SNARE complexes bound to the Synaptotagmin-1 C2B domain and complexin-1 in a spring-loaded configuration that prevents premature membrane merger and formation of extended interfaces, but keeps the system ready for fast fusion upon Ca2+ influx.

N-3-Oxo-Octanoyl Homoserine Lactone Primes Plant Resistance Against Necrotrophic Pathogen Pectobacterium carotovorum by Coordinating Jasmonic Acid and Auxin-Signaling Pathways.

Many Gram-negative bacteria use small signal molecules, such as N-acyl-homoserine lactones (AHLs), to communicate with each other and coordinate their collective behaviors. Recently, increasing evidence has demonstrated that long-chained quorum-sensing signals play roles in priming defense responses in plants. Our previous work indicated that a short-chained signal, N-3-oxo-octanoyl homoserine lactone (3OC8-HSL), enhanced Arabidopsis resistance to the hemi-biotrophic bacteria Pseudomonas syringae pv. tomato DC3000 through priming the salicylic acid (SA) pathway. Here, we found that 3OC8-HSL could also prime resistance to the necrotrophic bacterium Pectobacterium carotovorum ssp. carotovorum (Pcc) through the jasmonic acid (JA) pathway, and is dependent on auxin responses, in both Chinese cabbage and Arabidopsis. The subsequent Pcc invasion triggered JA accumulation and increased the down-stream genes’ expressions of JA synthesis genes (LOX, AOS, and AOC) and JA response genes (PDF1.2 and VSP2). The primed state was not observed in the Arabidopsis coi1-1 and jar1-1 mutants, which indicated that the primed resistance to Pcc was dependent on the JA pathway. The 3OC8-HSL was not transmitted from roots to leaves and it induced indoleacetic acid (IAA) accumulation and the DR5 and SAUR auxin-responsive genes’ expressions in seedlings. When Arabidopsis and Chinese cabbage roots were pretreated with exogenous IAA (10 μM), the plants had activated the JA pathway and enhanced resistance to Pcc, which implied that the JA pathway was involved in AHL priming by coordinating with the auxin pathway. Our findings provide a new strategy for the prevention and control of soft rot in Chinese cabbage and provide theoretical support for the use of the quorum-sensing AHL signal molecule as a new elicitor.

Characterization of monadic BL-algebras by state operators

Abstract In this paper, we introduce the notion of state monadic BL-algebras and we investigate their properties. We define the concept of state monadic filters and study certain types of state monadic filters; we define and charaterize maximal and prime state monadic filters. Characterizations of local, simple and semisimple state monadic BL-algebras are also explored in the paper as well.

A Comparative Study of Cell Culture Conditions during Conversion from Primed to Naive Human Pluripotent Stem Cells.

The successful reprogramming of human somatic cells into induced pluripotent stem cells (hiPSCs) represented a turning point in the stem cell research field, owing to their ability to differentiate into any cell type with fewer ethical issues than human embryonic stem cells (hESCs). In mice, PSCs are thought to exist in a naive state, the cell culture equivalent of the immature pre-implantation embryo, whereas in humans, PSCs are in a primed state, which is a more committed pluripotent state than a naive state. Recent studies have focused on capturing a similar cell stage in human cells. Given their earlier developmental stage and therefore lack of cell-of-origin epigenetic memory, these cells would be better candidates for further re-differentiation, use in disease modeling, regenerative medicine and drug discovery. In this study, we used primed hiPSCs and hESCs to evaluate the successful establishment and maintenance of a naive cell stage using three different naive-conversion media, both in the feeder and feeder-free cells conditions. In addition, we compared the directed differentiation capacity of primed and naive cells into the three germ layers and characterized these different cell stages with commonly used pluripotent and lineage-specific markers. Our results show that, in general, naive culture NHSM medium (in both feeder and feeder-free systems) confers greater hiPSCs and hESCs viability and the highest naive pluripotency markers expression. This medium also allows better cell differentiation cells toward endoderm and mesoderm.

Cell fate roadmap of human primed-to-naive transition reveals preimplantation cell lineage signatures

Human naive pluripotent stem cells offer a unique window into early embryogenesis studies. Recent studies have reported several strategies to obtain cells in the naive state. However, cell fate transitions and the underlying mechanisms remain poorly understood. Here, by a dual fluorescent reporter system, we depict the cell fate dynamics from primed state toward naive pluripotency with ALPG activation followed by the activation of OCT4-distal enhancer. Integration of transcription profiles and the chromatin accessibility landscape reveals the appearance of primitive endoderm and trophectoderm signatures in the transitioning subpopulations, with the capacities for derivation of extra-embryonic endoderm and trophoblast stem cell lines, respectively. Furthermore, despite different fluorescent dynamics, all transitioning intermediates are capable of reaching the naive state with prolonged induction, showing their developmental plasticity and potential. Overall, our study describes a global cell roadmap toward naive pluripotency and provides hints for embryo modeling-related studies.

ID proteins promote the survival and primed-to-naive transition of human embryonic stem cells through TCF3-mediated transcription

Inhibition of DNA binding proteins 1 and 3 (ID1 and ID3) are important downstream targets of BMP signalling that are necessary for embryonic development. However, their specific roles in regulating the pluripotency of human embryonic stem cells (hESCs) remain unclear. Here, we examined the roles of ID1 and ID3 in primed and naive-like hESCs and showed that ID1 and ID3 knockout lines (IDs KO) exhibited decreased survival in both primed and naive-like state. IDs KO lines in the primed state also tended to undergo pluripotent dissolution and ectodermal differentiation. IDs KO impeded the primed-to-naive transition (PNT) of hESCs, and overexpression of ID1 in primed hESCs promoted PNT. Furthermore, single-cell RNA sequencing demonstrated that ID1 and ID3 regulated the survival and pluripotency of hESCs through the AKT signalling pathway. Finally, we showed that TCF3 mediated transcriptional inhibition of MCL1 promotes AKT phosphorylation, which was confirmed by TCF3 knockdown in KO lines. Our study suggests that IDs/TCF3 acts through AKT signalling to promote survival and maintain pluripotency of both primed and naive-like hESCs.

Long-term priming of hypothalamic microglia is associated with energy balance disturbances under diet-induced obesity.

Exposure of microglia to an inflammatory environment may lead to their priming and exacerbated response to future inflammatory stimuli. Here we aimed to explore hypothalamic microglia priming and its consequences on energy balance regulation. A model of intracerebroventricular administration of neuraminidase (NA, which is present in various pathogens such as influenza virus) was used to induce acute neuroinflammation. Evidences of primed microglia were observed 3 months after NA injection, namely (1) a heightened response of microglia located in the hypothalamic arcuate nucleus after an in vivo inflammatory challenge (high fat diet [HFD] feeding for 10 days), and (2) an enhanced response of microglia isolated from NA‐treated mice and challenged in vitro to LPS. On the other hand, the consequences of a previous NA‐induced neuroinflammation were further evaluated in an alternative inflammatory and hypercaloric scenario, such as the obesity generated by continued HDF feeding. Compared with sham‐injected mice, NA‐treated mice showed increased food intake and, surprisingly, reduced body weight. Besides, NA‐treated mice had enhanced microgliosis (evidenced by increased number and reactive morphology of microglia) and a reduced population of POMC neurons in the basal hypothalamus. Thus, a single acute neuroinflammatory event may elicit a sustained state of priming in microglial cells, and in particular those located in the hypothalamus, with consequences in hypothalamic cytoarchitecture and its regulatory function upon nutritional challenges.

Abstract B028: Overcoming resistance by driving melanoma cells into drug-susceptible states

Abstract The goal of this study is to target pre-existing heterogeneity in gene expression to ultimately overcome resistance to targeted therapy in melanoma. Despite significant advances in the treatment of melanoma, many patients still fail to achieve durable responses to therapy due to the development of drug resistance. Our lab recently discovered that in drug-naive melanoma rare cells transiently enter distinct gene expression states that allows them to ultimately survive treatment with targeted therapy (we refer to these cells as “primed for drug resistance”). The goal of this work is to develop a strategy to overcome this form of resistance by driving cells out of states primed for resistance and into drug-susceptible states. To identify the pathways that regulate the transition to the primed state we combined single-cell RNA sequencing with DNA barcoding to extract the transcriptional profiles and lineage information of over 7,000 drug-naive melanoma cells. We found that cells in lineages that transitioned into states primed for drug resistance express high levels of genes associated with TGFβ and PI3K signaling compared to lineages that stayed in a drug-susceptible state. To test whether TGFβ can directly cause cells to enter the primed state, we treated cells with recombinant TGFβ which led to a significant increase in the number of primed cells. Intriguingly, we also found that treating with TGFB receptor inhibitor or PI3K inhibitor reduced the number of cells in the primed cell state. Further, we sequentially treated these cells first with either the TGFB or PI3K inhibitor followed by targeted therapy and ultimately reduced the number of resistant colonies, demonstrating the potential for this therapeutic paradigm. To better understand how targeting the TGFB and PI3K pathways shift the population dynamics, we used single-cell RNA-seq and cellular barcoding to track lineages following treatment with TGFB, TGFB inhibitor, and PI3K inhibitor. Our analysis reveals the population changes following each therapy and directly demonstrates that treating with PI3K inhibitor decreases the number of primed cells, shifting the population into a drug-susceptible expression state. Ultimately, this work defines two key pathways underlying the primed cell state in melanoma and presents a therapeutic paradigm to eliminate this form of therapy resistance. Citation Format: Guillaume Harmange, Ben Emert, Dylan Schaff, Raul Reyes-Hueros, Sydney Shaffer. Overcoming resistance by driving melanoma cells into drug-susceptible states [abstract]. In: Proceedings of the AACR Special Conference on the Evolutionary Dynamics in Carcinogenesis and Response to Therapy; 2022 Mar 14-17. Philadelphia (PA): AACR; Cancer Res 2022;82(10 Suppl):Abstract nr B028.

Untangling the Web of Protein Kinase C Mediated Regulation of Naïve Vs Primed State of Pluripotency

Introduction‐Pluripotency of stem cells is associated with the dynamicity of the embryo’s developmental stage from which they are derived. Stem cells derived from the pre‐implantation embryo, called embryonic stem cells (ESCs), are naïve in nature, whereas post‐implantation embryo‐derived stem cells, termed epiblast stem cells (EpiSCs), represent the primed state of pluripotency. Maintaining these stem cells in a naïve state is an essential arena of stem cell research since naïve ESCs are capable of clonal propagation and germline transmission, making them a crucial research tool in organogenesis and disease modeling. Inhibition of PKC (PKCi) signaling was found to maintain an undifferentiated stage of pluripotency in both mouse and rat. Interestingly, when used along with a cocktail of inhibitors, the same inhibitor facilitated the derivation of naïve state pluripotency in human ESC. These findings lead us to a hypothesis that PKC signaling inhibition alone has a role in maintaining the naïve state of pluripotency. Hence, it has intrigued us to explore the complex network of mechanisms exerted by PKC signaling in dictating the transition of naïve to primed state of pluripotency.Methodology‐In that pursuit, we first observed the morphology of the mouse ESCs (mESCs) by phase‐contrast microscopy across passages and performed MTT assay to optimize the concentration used for further assay. Quantitative Real‐time PCR and western blotting were performed to determine the expression of transcription factors associated with naïve and primed states of pluripotency. Other techniques, including FACS, TiO2 phospho‐enrichment followed by Mass‐spectrometric (MS) analysis, Immunofluorescence assay, Immunoprecipitation assay, Nuclear‐cytoplasmic protein extraction and others were exploited to study the mechanism.Results‐We observed that PKCi could maintain naïve state of pluripotency in mESC even after prolonged culturing by reducing the expression of a critical primed transcription factor. The inhibition of phosphorylation of the PKCζ isoform is responsible for naïve state maintenance. Cell cycle analysis showed that more cells are in the G1 phase in the PKCi maintained condition than in the cells maintained in the presence of Leukemia Inhibitory Factor (LIF). Also, cell proliferation marker CDK1 expression was less in PKCi maintained cells, which probably is negatively regulated by the transcription factor KLF4. Increased nuclear expression of KLF4 was observed in PKCi maintained ESCs than that of ESCs cultured with LIF. Beside these transcription factor‐mediated regulations, PKCi exerts an epigenetic control over the ESCs absent in LIF maintained ESCs. PKCi mediated loss in phosphorylation of the epigenetic factor contributes to naïve state maintenance. Further, our data indicates towards PKCζ mediated NF‐κβ pathway activation, thereby dictating the transition of naïve to primed state of pluripotency.Conclusions‐PKCi maintained mESCs could maintain the naïve state of pluripotency. We have unraveled a multifaceted functionality of PKC signaling in the transition of naïve to primed state of pluripotency, which extends to both transcriptional and epigenetic regulation.

Long-Lasting Defence Priming by β-Aminobutyric Acid in Tomato Is Marked by Genome-Wide Changes in DNA Methylation.

Exposure of plants to stress conditions or to certain chemical elicitors can establish a primed state, whereby responses to future stress encounters are enhanced. Stress priming can be long-lasting and likely involves epigenetic regulation of stress-responsive gene expression. However, the molecular events underlying priming are not well understood. Here, we characterise epigenetic changes in tomato plants primed for pathogen resistance by treatment with β-aminobutyric acid (BABA). We used whole genome bisulphite sequencing to construct tomato methylomes from control plants and plants treated with BABA at the seedling stage, and a parallel transcriptome analysis to identify genes primed for the response to inoculation by the fungal pathogen, Botrytis cinerea. Genomes of plants treated with BABA showed a significant reduction in global cytosine methylation, especially in CHH sequence contexts. Analysis of differentially methylated regions (DMRs) revealed that CHH DMRs were almost exclusively hypomethylated and were enriched in gene promoters and in DNA transposons located in the chromosome arms. Genes overlapping CHH DMRs were enriched for a small number of stress response-related gene ontology terms. In addition, there was significant enrichment of DMRs in the promoters of genes that are differentially expressed in response to infection with B. cinerea. However, the majority of genes that demonstrated priming did not contain DMRs, and nor was the overall distribution of methylated cytosines in primed genes altered by BABA treatment. Hence, we conclude that whilst BABA treatment of tomato seedlings results in characteristic changes in genome-wide DNA methylation, CHH hypomethylation appears only to target a minority of genes showing primed responses to pathogen infection. Instead, methylation may confer priming via in-trans regulation, acting at a distance from defence genes, and/or by targeting a smaller group of regulatory genes controlling stress responses.

Chemical Pretreatment Activated a Plastic State Amenable to Direct Lineage Reprogramming.

Somatic cells can be chemically reprogrammed into a pluripotent stem cell (CiPSC) state, mediated by an extraembryonic endoderm- (XEN-) like state. We found that the chemical cocktail applied in CiPSC generation initially activated a plastic state in mouse fibroblasts before transitioning into XEN-like cells. The plastic state was characterized by broadly activated expression of development-associated transcription factors (TFs), such as Sox17, Ascl1, Tbx3, and Nkx6-1, with a more accessible chromatin state indicating an enhanced capability of cell fate conversion. Intriguingly, introducing such a plastic state remarkably improved the efficiency of chemical reprogramming from fibroblasts to functional neuron-like cells with electrophysiological activity or beating skeletal muscles. Furthermore, the generation of chemically induced neuron-like cells or skeletal muscles from mouse fibroblasts was independent of the intermediate XEN-like state or the pluripotency state. In summary, our findings revealed a plastic chemically activated multi-lineage priming (CaMP) state at the onset of chemical reprogramming. This state enhanced the cells’ potential to adapt to other cell fates. It provides a general approach to empowering chemical reprogramming methods to obtain functional cell types bypassing inducing pluripotent stem cells.

Activating stress memory: eustressors as potential tools for plant breeding.

Plants are continuously exposed to stress conditions, such that they have developed sophisticated and elegant survival strategies, which are reflected in their phenotypic plasticity, priming capacity, and memory acquisition. Epigenetic mechanisms play a critical role in modulating gene expression and stress responses, allowing malleability, reversibility, stability, and heritability of favourable phenotypes to enhance plant performance. Considering the urgency to improve our agricultural system because of going impacting climate change, potential and sustainable strategies rely on the controlled use of eustressors, enhancing desired characteristics and yield and shaping stress tolerance in crops. However, for plant breeding purposes is necessary to focus on the use of eustressors capable of establishing stable epigenetic marks to generate a transgenerational memory to stimulate a priming state in plants to face the changing environment.

Novel sustainable strategies to control Plasmopara viticola in grapevine unveil new insights on priming responses and arthropods ecology.

Reduction of fungicide consumption in agriculture is globally recognized as a priority. Government authorities are fostering research to achieve a reduction of risks associated with conventional pesticides and promoting the development of sustainable alternatives. To address these issues, in the present study, alternative protocols for the control of downy mildew infection in grapevine were compared to the standard protocol. In the first protocol, only resistance inducers were used, comprising a single formulation with Acibenzolar S-methyl, laminarin and disodium-phosphonate. The second and third protocols followed the standard protocol but substituted phosphonates with phosphorus pentoxide and Ecklonia maxima extract. The results showed that at veraison downy mildew incidence and severity in all tested protocols were significantly reduced compared to nontreated controls on both canopy and bunches. Expression analysis of key genes involved in plant stress response, indicated that the two protocols for phosphites substitution induced a remodulation of salicylic acid (SA) and jasmonic acid (JA), with positive impact on yields. Analysis of the first protocol revealed that the primed state induced a short delay in bunch ripening, with a shift of carbohydrate metabolism to boost the plant defences, involving an upregulation of defence related-gene, SAR response and a decreased ROS detoxification. Additionally, analysis on the arthropods populations, in parallel with the positive results achieved using alternatives to conventional fungicides, were enriched by those showing the potential of naturally occurring predators of spider mites. This study provides practical solutions to reduce the environmental impact of treatments for the control downy mildew in viticulture. © 2022 Society of Chemical Industry.

Decoding the potential of a new Pseudomonas putida strain for inducing drought tolerance of tomato (Solanum lycopersicum) plants through seed biopriming

A total of 11 potential plant growth promoting rhizobacteria previously isolated from naturally stressed environments were evaluated for various traits of interest for a beneficial symbiosis with plants, including colonization ability, biofilm formation, motility, exopolysaccharide production and salt tolerance. The vast majority of the strains were found to possess multiple plant growth promoting traits. Nevertheless, the intensity varied among isolates, with those originated from tomato plants being more efficient colonizers. The strain SAESo11, genetically characterized as a Pseudomonas putida member was selected for further investigation of its potential to alleviate drought stress in tomato seedlings. Inoculation with SAESo11 mitigated the negative effects of drought stress as indicated by growth and photosynthetic indices. Furthermore, bacterial inoculation enhanced H2O2 content and malondialdehyde levels in colonized plants. Drought treatment did not further alter the oxidative status of these plants. Similarly, total phenolic content and antioxidant enzyme activity were induced in plant tissues in response to drought stress only at the absence of inoculum. These results indicated that inoculation with the selected strain imposed plants at a priming state, that enabled them to respond more robustly at the exposure to drought stress and efficiently attenuated the drought-induced injury. This state of plant alertness mediated by SAESo11 occurred at no cost to growth, highlighting its role as a potential plant priming agent.

Transcriptome Responses of Wild Arachis to UV-C Exposure Reveal Genes Involved in General Plant Defense and Priming.

Stress priming is an important strategy for enhancing plant defense capacity to deal with environmental challenges and involves reprogrammed transcriptional responses. Although ultraviolet (UV) light exposure is a widely adopted approach to elicit stress memory and tolerance in plants, the molecular mechanisms underlying UV-mediated plant priming tolerance are not fully understood. Here, we investigated the changes in the global transcriptome profile of wild Arachis stenosperma leaves in response to UV-C exposure. A total of 5751 differentially expressed genes (DEGs) were identified, with the majority associated with cell signaling, protein dynamics, hormonal and transcriptional regulation, and secondary metabolic pathways. The expression profiles of DEGs known as indicators of priming state, such as transcription factors, transcriptional regulators and protein kinases, were further characterized. A meta-analysis, followed by qRT-PCR validation, identified 18 metaDEGs as being commonly regulated in response to UV and other primary stresses. These genes are involved in secondary metabolism, basal immunity, cell wall structure and integrity, and may constitute important players in the general defense processes and establishment of a priming state in A. stenosperma. Our findings contribute to a better understanding of transcriptional dynamics involved in wild Arachis adaptation to stressful conditions of their natural habitats.

Selective Boosting of CCR7-Acting Chemokines; Short Peptides Boost Chemokines with Short Basic Tails, Longer Peptides Boost Chemokines with Long Basic Tails.

The chemokine receptor CCR7 and its ligands CCL19 and CCL21 regulate the lymph node homing of dendritic cells and naïve T-cells and the following induction of a motile DC-T cell priming state. Although CCL19 and CCL21 bind CCR7 with similar affinities, CCL21 is a weak agonist compared to CCL19. Using a chimeric chemokine, CCL19CCL21N-term|C-term, harboring the N-terminus and the C-terminus of CCL21 attached to the core domain of CCL19, we show that these parts of CCL21 act in a synergistic manner to lower ligand potency and determine the way CCL21 engages with CCR7. We have published that a naturally occurring basic C-terminal fragment of CCL21 (C21TP) boosts the signaling of both CCL19 and CCL21. Boosting occurs as a direct consequence of C21TP binding to the CCR7 N-terminus, which seems to free chemokines with basic C-termini from an unfavorable interaction with negatively charged posttranslational modifications in CCR7. Here, we confirm this using a CCL19-variant lacking the basic C-terminus. This variant displays a 22-fold higher potency at CCR7 compared to WT CCL19 and is highly unaffected by the presence of C21TP. WT CCL19 has a short basic C-terminus, CCL21 a longer one. Here, we propose a way to differentially boost CCL19 and CCL21 activity as short and long versions of C21TP boost CCL19 activity, whereas only a long C21TP version can boost chemokines with a full-length CCL21 C-terminus.

Advances in 5-Aminolevulinic Acid Priming to Enhance Plant Tolerance to Abiotic Stress.

Priming is an adaptive strategy that improves plant defenses against biotic and abiotic stresses. Stimuli from chemicals, abiotic cues, and pathogens can trigger the establishment of priming state. Priming with 5-aminolevulinic acid (ALA), a potential plant growth regulator, can enhance plant tolerance to the subsequent abiotic stresses, including salinity, drought, heat, cold, and UV-B. However, the molecular mechanisms underlying the remarkable effects of ALA priming on plant physiology remain to be elucidated. Here, we summarize recent progress made in the stress tolerance conferred by ALA priming in plants and provide the underlying molecular and physiology mechanisms of this phenomenon. Priming with ALA results in changes at the physiological, transcriptional, metabolic, and epigenetic levels, and enhances photosynthesis and antioxidant capacity, as well as nitrogen assimilation, which in turn increases the resistance of abiotic stresses. However, the signaling pathway of ALA, including receptors as well as key components, is currently unknown, which hinders the deeper understanding of the defense priming caused by ALA. In the future, there is an urgent need to reveal the molecular mechanisms by which ALA regulates plant development and enhances plant defense with the help of forward genetics, multi-omics technologies, as well as genome editing technology.

Germ cells commit somatic stem cells to differentiation following priming by PI3K/Tor activity in the Drosophila testis.

How and when potential becomes restricted in differentiating stem cell daughters is poorly understood. While it is thought that signals from the niche are actively required to prevent differentiation, another model proposes that stem cells can reversibly transit between multiple states, some of which are primed, but not committed, to differentiate. In the Drosophila testis, somatic cyst stem cells (CySCs) generate cyst cells, which encapsulate the germline to support its development. We find that CySCs are maintained independently of niche self-renewal signals if activity of the PI3K/Tor pathway is inhibited. Conversely, PI3K/Tor is not sufficient alone to drive differentiation, suggesting that it acts to license cells for differentiation. Indeed, we find that the germline is required for differentiation of CySCs in response to PI3K/Tor elevation, indicating that final commitment to differentiation involves several steps and intercellular communication. We propose that CySC daughter cells are plastic, that their fate depends on the availability of neighbouring germ cells, and that PI3K/Tor acts to induce a primed state for CySC daughters to enable coordinated differentiation with the germline.

Transcriptomic Analysis of Human Naïve and Primed Pluripotent Stem Cells.

Over the last decade, RNA-Sequencing (RNA-Seq) has revolutionized the field of transcriptomics due to its sheer advantage over previous technologies for studying gene expression. Even the domain of stem cell bioinformatics has benefited from these advancements. It has helped look deeper into how the process of pluripotency is maintained by stem cells and how it may be exploited for application in regenerative medicine. However, as it is still an evolving technology, there is no single accepted protocol for RNA-Seq data analysis. From a wide array of tools and/or algorithms available for the purpose, researchers tend to develop a pipeline that is best suited for their sample, experimental design, and computational power. In this tutorial, we describe a pipeline based on open-source tools to analyze RNA-Seq data from naïve and primed state human pluripotent stem cell samples. Precisely, we show how RNA-Seq data can be downloaded from databases, processed, and used to identify differentially expressed genes and construct a co-expression network. Further, we also show how the list of interesting genes obtained from differential expression testing or co-expression network be analyzed to gain biological insights.