Nuclear Molecules Research Articles

P0167 Circular extrachromosomal DNA (eccDNA): a potential diagnostic and prognostic tool to be introduced into the clinical practice of Inflammatory Bowel Disease

Abstract Background The diagnosis of Inflammatory Bowel Disease, which continues to increase year by year, is complex; these disorders can be confirmed by investigations such as fecal calprotectin, endoscopy, histology of biopsies, and CT scan; despite this, there is a lack of highly specific and minimally invasive techniques that allow not only diagnosis but also determination of the severity of the disease and its evolution. Given these challenges, the search for reliable and specific predictors that can direct the clinician toward personalized patient care is very important. In this context, can help extrachromosomal circular DNA (eccDNA) that are eukaryotic nuclear circle DNA molecules, independent from chromosomes, with a length ranging from a few hundred to a few thousand base pairs. In addition to its already known role in cancer, in IBD increased levels of eccDNA have been found compared with healthy controls. Circular DNA could represent a potential biomarker that can be isolated from peripheral blood or intestinal mucosa and is useful for making early diagnoses and assessing disease status. Methods We examined peripheral blood, collected by blood sampling . "Technical Tests" were performed in order to determine the right procedure for storing blood samples and optimize the results obtained in terms of eccDNA extraction . The samples will be divided into two groups, for the first group the procedure is to centrifuge the blood at 2000g for 15 minutes at different timepoints with or without storing the plasma at 4° C; for the second group the sample is centrifuged immediately after blood collection and the plasma is stored in the refrigerator at different timepoints [Table 1]. Then for both groups we proceed with eccDNA extraction by the "Circle-Seq" analysis. Results No significant differences are shown between the two groups in terms of the amount of eccDNA [Figure 1]. In particular, the non-immediate centrifugation and separation of plasma from whole blood does not negatively affect the extraction yield, highlighting that degradative factors are not present in whole blood. In addition, refrigerated storage for a longer period of time is a positive factor for eccDNA concentration, probably because in this way circular DNA found within microvesicles has time to be extruded, increasing its concentration in plasma. Conclusion Circulating eccDNAs could represent potential biomarkers to be detected in plasma by liquid biopsy, as the method is minimally invasive and easily reproducible in analytical laboratories, becoming a diagnostic and prognostic tool to be brought into clinical practice. References Fundings: European Union’s Horizon 2020 Research and Innovation Programme (899417)

The secrets of the Tübingen Castle kitchen: Friedrich Miescher and the discovery of nuclein, the cornerstone of DNA.

In 1869, Friedrich Miescher, born in Basel, Switzerland, discovered a previously unknown phosphorus-rich substance in the nuclei of pus cells. Conducting his research in a laboratory set up in the kitchen of Tübingen's medieval castle in Germany, and under the guidance by Professor Felix Hoppe-Seyler, Miescher primarily focused on the composition of cell nuclei. He obtained nuclear material by washing pus cells from surgical bandages provided by a nearby hospital. In 1869, Miescher described a completely new nuclear molecule that, unlike proteins, contained large amounts of phosphorus. He named this substance 'nuclein,' marking the first description of DNA. Miescher passed away long before Watson, Crick, Wilkins, and Franklin announced the precise role DNA played in cells in 1953. Through his discovery, Miescher laid the groundwork for all subsequent DNA research.

Nuclear-Targeting Peptides for Cancer Therapy.

Nucleus is the central regulator of cells that controls cell proliferation, metabolism, and cell cycle, and is considered the most important organelle in cells. The precision medicine that can achieve nuclear targeting has achieved good therapeutic effects in anti-tumor therapy. However, the presence of biological barriers such as cell membranes and nuclear membranes in cells limit the delivery of therapeutic agents to the nucleus. Therefore, developing effective nuclear-targeting drug delivery strategies is particularly important. Nuclear-targeting peptides are a class of functional peptides that can penetrate cell membranes and target the nucleus. They mainly recognize and bind to the nuclear transport molecules (such as Importin-α/β) and transport the therapeutic agents to the nucleus through nuclear pore complexes (NPC). This review summarizes the most recent developments of strategies for anti-tumor therapy utilizing nuclear-targeting peptides, which will ultimately contribute to the development of more effective nuclear-targeting strategies to achieve better anti-tumor outcomes.

The Integrator complex: an emerging complex structure involved in the regulation of gene expression by targeting RNA polymerase II.

The Integrator complex is a multisubunit complex that participates in the processing of small nuclear RNA molecules in eukaryotic cells by cleaving the 3' end. In protein-coding genes, Integrator is a key regulator of promoter-proximal pausing, release, and recruitment of RNA polymerase II. Research on Integrator has revealed its critical role in the regulation of gene expression and RNA processing. Dysregulation of the Integrator complex has been implicated in a variety of human diseases including cancer and developmental disorders. Therefore, understanding the structure and function of the Integrator complex is critical to uncovering the mechanisms of gene expression and developing potential therapeutic strategies for related diseases.

Unique Interplay Between Antinuclear Antibodies and Nuclear Molecules in the Pathogenesis of Systemic Lupus Erythematosus.

Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease that primarily affects young women and causes a wide range of inflammatory manifestations. The hallmark of SLE is the production of antibodies to components of the cell nucleus (antinuclear antibodies [ANAs]). These antibodies can bind to DNA, RNA, and protein complexes with nucleic acids. Among ANAs, antibodies to DNA (anti-DNA) are markers for classification and disease activity, waxing and waning disease activity in many patients. In the blood, anti-DNA antibodies can bind to DNA to form immune complexes with two distinct roles in pathogenesis: (1) renal deposition to provoke nephritis and (2) stimulation of cytokine production following uptake into innate immune cells and interaction with internal nucleic acid sensors. These sensors are part of an internal host defense system in the cell cytoplasm that can respond to DNA from infecting organisms; during cell stress, DNA from nuclear and mitochondrial sources can also trigger these sensors. The formation of immune complexes requires a source of extracellular DNA in an immunologically accessible form. As shown in in vivo and in vitro systems, extracellular DNA can emerge from dead and dying cells in both a free and a particulate form. Neutrophils undergoing the process of NETosis can release DNA in mesh-like structures called neutrophil extracellular traps. In SLE, therefore, the combination of ANAs and immunologically active DNA can create new structures that can promote inflammation throughout the body as well as drive organ inflammation and damage.

Cluster effects on low-energy carbon burning

Carbon burning is crucial for the fate of massive stars and element creation in the Universe. Its understanding at stellar energies is a great theoretical challenge, as carbon fusion is a multidimensional quantum tunneling process affected by resonances whose origin is not fully understood yet. Simplified quantum dynamical coupled-channels calculations are presented, suggesting that clustering in the intermediate nuclear molecule is a source of fusion resonances.

Proteomic Profiling of Early Secreted Proteins in Response to Lipopolysaccharide-Induced Vascular Endothelial Cell EA.hy926 Injury.

Sepsis is a crucial public health problem with a high mortality rate caused by a dysregulated host immune response to infection. Vascular endothelial cell injury is an important hallmark of sepsis, which leads to multiple organ failure and death. Early biomarkers to diagnose sepsis may provide early intervention and reduce risk of death. Damage-associated molecular patterns (DAMPs) are host nuclear or cytoplasmic molecules released from cells following tissue damage. We postulated that DAMPs could potentially be a novel sepsis biomarker. We used an in vitro model to determine suitable protein-DAMPs biomarkers for early sepsis diagnosis. Low and high lipopolysaccharide (LPS) doses were used to stimulate the human umbilical vein endothelial cell line EA.hy926 for 24, 48, and 72 h. Results showed that cell viability was reduced in both dose-dependent and time-dependent manners. Cell injury was corroborated by a significant increase in lactate dehydrogenase (LDH) activity within 24 h in cell-conditioned medium. Secreted protein-DAMPs in the supernatant, collected at different time points within 24 h, were characterized using shotgun proteomics LC-MS/MS analysis. Results showed that there were 2233 proteins. Among these, 181 proteins from the LPS-stimulated EA.hy926 at 1, 12, and 24 h were significantly different from those of the control. Twelve proteins were up-regulated at all three time points. Furthermore, a potential interaction analysis of predominant DAMPs-related proteins using STITCH 5.0 revealed the following associations with pathways: response to stress; bacterium; and LPS (GO:0080134; 0009617; 0032496). Markedly, alpha-2-HS-glycoprotein (AHSG or fetuin-A) and lactotransferrin (LTF) potentially presented since the first hour of LPS stimulation, and were highly up-regulated at 24 h. Taken together, we reported proteomic profiling of vascular endothelial cell-specific DAMPs in response to early an in vitro LPS stimulation, suggesting that these early damage-response protein candidates could be novel early biomarkers associated with sepsis.

Role of crustacean female sex hormone in regulating immune response in the mud crab, Scylla paramamosain

Crustacean female sex hormone (CFSH) is responsible for sexual differentiation in crustaceans. The CFSH exhibited an interleukin-17 domain homologous to vertebrate IL-17, a family of inflammatory cytokines that play vital roles in immune defense. However, the immunoregulation of CFSH in crustaceans is a mystery. Therefore, this study aimed to investigate the immune regulatory roles of CFSH and CFSHR in the mud crab Scylla paramamosain. This study's immunofluorescence result revealed that Sp-CFSHR was highly expressed in granulocytes and semi-granulocytes but had moderate expression in hyalinocytes. The expression level of Sp-CFSH transcript in eyestalk ganglia and Sp-CFSHR in hemocytes were significantly up-regulated by the Poly (I:C) stimulation but significantly down-regulated in response to the lipopolysaccharide (LPS) stimulation. In our study, in vitro experiment exhibited that the nuclear transcription factors NF-κB signaling molecules (Sp-Dorsal and Sp-Relish), Sp-STAT, apoptosis-related gene Sp-IAP, and phagocytosis related gene (Sp-Rab5) expressions were significantly increased in hemocytes by recombinant CFSH (rCFSH) in vitro, but the pro-inflammatory cytokine gene (Sp-IL-16) expression was significantly suppressed. Finally, the rCFSH injection significantly up-regulated Sp-Dorsal, Sp-Relish, Sp-IAP, Sp-Caspase, Sp-ALF2, and C-type lectin (Sp-CTL-B) expressions in hemocytes as well as enhanced the bacterial clearance of the mud crab. In conclusion, our results suggested that CFSH may be a counterpart of vertebrate IL-17 in crustaceans that can enhance innate immunity to defense against Vibrionaceae infection via the NF-κB and/or JAK-STAT signaling pathways. This study provides the first evidence that CFSH is involved in the immunoregulation in crustaceans and enriches the insight of neuroendocrine-immune regulatory system, which providing new ideas for disease prevention in the mud crab industry.

2D approach to reconstruct the evolutionary history of clevelandellids (Ciliophora, Armophorea) inhabiting the hindgut of the Panesthiinae cockroaches

AbstractThe ciliate order Clevelandellida unites endosymbionts of the digestive tract of a variety of invertebrates and vertebrates. In the present study, the primary and secondary structures of nuclear and hydrogenosomal rRNA molecules were employed to reconstruct the phylogenetic relationships and to estimate the divergence times of clevelandellids inhabiting the hindgut of the Panesthiinae cockroaches. The secondary structure information was incorporated in phylogenetic analyses using two different strategies, viz., indirectly through 2D‐guided alignments and directly through so‐called pseudo‐protein data. Nuclear and hydrogenosomal markers carried a consistent phylogenetic signal and robustly supported the monophyletic origin of the family Clevelandellidae as well as of its four genera. According to Bayesian relaxed molecular clock analyses, the last common ancestor (LCA) of the family Clevelandellidae very likely emerged during the Late Cretaceous in the Oriental region. Its descendants most likely expanded to Australia in concert with the Neogene colonization and radiation of their host Panesthiinae cockroaches. Taking into account the time‐calibrated phylogenies and the fact that early branching members of the order Clevelandellida inhabit the digestive tracts of amphibians, it is tempting to speculate that the LCA of the Clevelandellida evolved in ectothermic vertebrates. Amphibians could have brought clevelandellids to the land, where they may have been transmitted to the cockroach digestive tract upon feeding on amphibian faeces.

Bringing to light nuclear-mitochondrial insertions in the genomes of nocturnal predatory birds

Mito-nuclear insertions, or NUMTs, relate to genetic material of mitochondrial origin that have been transferred to the nuclear DNA molecule. The increasing amounts of genomic data currently being produced presents an opportunity to investigate this type of patterns in genome evolution of non-model organisms. Identifying NUMTs across a range of closely related taxa allows one to generalize patterns of insertion and maintenance in autosomes, which is ultimately relevant to the understanding of genome biology and evolution. Here we collected existing pairwise genome-mitogenome data of the order Strigiformes, a group that includes all the nocturnal bird predators. We identified NUMTs by applying percent similarity thresholds after blasting mitochondrial genomes against nuclear genome assemblies. We identified NUMTsin all genomes with numbers ranging from 4 in Bubo bubo to 24 in Ciccaba nigrolineata. Statistical analyses revealed NUMT size to negatively correlate with NUMT’s sequence similarity to with original mtDNA region. Lastly, characterizing these nuclear insertions of mitochondrial origin in a comparative genomics framework produced variable phylogenetic patterns, suggesting in some cases that insertions might pre-date speciation events within Strigiformes.

STORM Microscopy and Cluster Analysis for PcG Studies.

Advanced microscopy techniques (such as STORM, STED, andSIM) have recently allowed the visualization of biological samples beyond the diffraction limit of light. Thanks to this breakthrough, the organization of molecules can be revealed within single cells as never before.Here, we describe the application of STochastic Optical Reconstruction Microscopy (STORM) for the study of polycomb group of proteins (PcG) in the context of chromatin organization. We present a clustering algorithm to quantitatively analyze the spatial distribution of nuclear molecules (e.g., EZH2 or its associated chromatin mark H3K27me3) imaged by 2D STORM. This distance-based analysis uses x-y coordinates of STORM localizations to group them into "clusters." Clusters are classified as singles if isolated or into islands if they form a group of closely associated clusters. For each cluster, the algorithm calculates the number of localizations, the area, and the distance to the closest cluster.This approach can be used for every type of adherent cell lineand allows the imaging of every protein for which an antibody is available. It represents a comprehensive strategy to visualize and quantify how PcG proteins and related histone marks organize in the nucleus at nanometric resolution.

First molecular evidence of hybridization in endosymbiotic ciliates (Protista, Ciliophora).

Hybridization is an important evolutionary process that can fuel diversification via formation of hybrid species or can lead to fusion of previously separated lineages by forming highly diverse species complexes. We provide here the first molecular evidence of hybridization in wild populations of ciliates, a highly diverse group of free-living and symbiotic eukaryotic microbes. The impact of hybridization was studied on the model of Plagiotoma, an obligate endosymbiont of the digestive tube of earthworms, using split decomposition analyses and species networks, 2D modeling of the nuclear rRNA molecules and compensatory base change analyses as well as multidimensional morphometrics. Gene flow slowed down and eventually hampered the diversification of Lumbricus-dwelling plagiotomids, which collapsed into a single highly variable biological entity, the P. lumbrici complex. Disruption of the species boundaries was suggested also by the continuum of morphological variability in the phenotypic space. On the other hand, hybridization conspicuously increased diversity in the nuclear rDNA cistron and somewhat weakened the host structural specificity of the P. lumbrici complex, whose members colonize a variety of phylogenetically closely related anecic and epigeic earthworms. By contrast, another recorded species, P. aporrectodeae sp. n., showed no signs of introgression, no variability in the rDNA cistron, and very high host specificity. These contrasting eco-evolutionary patterns indicate that hybridization might decrease the alpha-diversity by dissolving species boundaries, weaken the structural host specificity by broadening ecological amplitudes, and increase the nuclear rDNA variability by overcoming concerted evolution within the P. lumbrici species complex.

Application of the cranking method to the semimicroscopic algebraic cluster model and nuclear molecules

A particular quantum phase transition (QPT) is studied at excited energies of light nuclei within the Semimicroscopic Algebraic Cluster Model (SACM), using a combination of catastrophe theory and a direct minimization of the potential. A distinct change from a compact nucleus to a nuclear molecule is described as a second order QPT occurring at a certain value of angular momentum. This finding is in accordance with experimental observations when nuclear molecules appear. The method has implication in the fission of heavy nuclei.

VP.75 Jab1 deletion in muscle lineage causes a muscular dystrophy that resembles LAMA2 disease

Merosin deficient congenital muscular dystrophy (LAMA2 disease) is an autosomal recessive disorder characterized by progressive wasting muscular dystrophy, dysmyelinating neuropathy and brain abnormalities. This disorder is caused by mutations in the <i>LAMA2</i> gene, which encodes for the laminin211, the primary component of the Schwann cell and muscle basal lamina. We recently identified Jab1 as a nuclear molecule downstream the laminin211 pathway and showed that Jab1 regulates Schwann cell behavior through the modulation of p27Kip1 levels. Interestingly, loss of Jab1 in Schwann cells results in a dysmyelinating neuropathy that phenocopies the neuropathy of LAMA2 disease. To evaluate whether Jab1 also plays a role in the pathogenesis of LAMA2 muscular dystrophy, we generated mice with conditional inactivation of <i>Jab1</i> in the muscle lineage through the MyoDi-cre transgene. Mice with deletion of <i>Jab1</i> in skeletal muscle (named Jab1-MscKO) showed progressive motor deficits, reduced lifespan and overt muscular dystrophy phenotype. Muscles appeared smaller in size and presented myopathic features such as reduced fiber diameter, presence of necrotic fibers, inflammatory cells, and fibrosis. The evaluation of satellite cell number showed a reduction of proliferating satellite cells, suggesting a defect of cell cycle progression. Muscle regeneration, induced by cardiotoxin injection, was significantly impaired in Jab1-MscKO mice. Jab1-MscKO muscles showed increased of p27Kip1 expression. Genetic deletion of <i>p27Kip1</i> in muscles of Jab1-MscKO mice revealed a mild, but not significant, amelioration of motor performances and muscle pathology, while reduced inflammatory infiltrates and ameliorated satellite cells cycling. Our results suggest that Jab1 in involved in the pathogenesis of LAMA2 muscular dystrophy, while p27Kip1 might influence the correct cell cycle progression of satellite cells.

Metabolomics of Meat Color: Practical Implications

Objective: Meat is biochemically active. Various pre-and post-harvest processes affect meat quality. Metabolomics is a valuable tool to elucidate metabolite changes in meat. The overall goal of this mini-review was to provide an overview of various techniques, data analysis, and application of metabolomics in meat color research. Results: Both targeted and non-targeted approaches are used to determine metabolite profiles in meat. Researchers use gas-, liquid-chromatography, and nuclear magnetic resonance platforms to separate molecules. Metabolomics is used to characterize muscle-specific differences in color stability, meat tenderness, the impact of aging on meat color, and to determine metabolite profile differences between normal-pH and dark-cutting beef. Color stable muscles have more glycolytic metabolites than color labile muscles. Conclusion: The use of metabolomics has greatly enhanced our understanding of metabolites' role in meat quality. There is a need for multiple databases to obtain comprehensive metabolite libraries specific to food. Metabolomics in combination with wet-laboratory techniques can provide novel insights on the relationship between postmortem metabolism and meat color.

Polarization of human gingival fibroblasts by Th1-, Th2-, Th17-, and Treg-derived cytokines.

The purpose of this study was to evaluate whether gingival fibroblasts (GFs) can be differently activated and polarized into distinct functional subtypes by T-helper (Th) cytokines. Gingival fibroblasts were stimulated with interferon (IFN)-γ, interleukin (IL)-4, IL-17, and transforming growth factor (TGF)-β, representative cytokines of Th1, Th2, Th17, and regulatory T cells, respectively, and the gene expression profiles were analyzed by microarray. Differentially expressed genes (DEGs) in GFs stimulated by 4 cytokines were screened, and a gene ontology (GO) analysis of the DEGs was conducted. To confirm the reliability of the microarray results, the DEGs that showed the largest differences compared with non-stimulated GFs were further analyzed by RT-PCR. To evaluate the effect of polarization on GFs responses to lipopolysaccharide (LPS), GFs stimulated by 4 cytokines were further stimulated with Escherichia coli LPS and mRNA levels of several genes were analyzed using RT-PCR. Differentially expressed genes by 4Th cytokines were enriched in different GO terms, and the patterns of gene expression on GFs were shown functionally different. GFs stimulated with IFN-γ (GF(IFN-γ)) up-regulated the expression of chemokines (chemokine (C-X-C motif) ligand (CXCL)9, -10, -11, chemokine (C-C motif) ligand (CCL)8), molecules involved in antigen presentation, complement component 3 (C3), and other immune response-related molecules, whereas they down-regulated the expression of several types of collagen, extracellular matrix (ECM) components, and DNA replication and nuclear protein-related molecules. By contrast, GF(IL-4) up-regulated the expression of ECM components, cell adhesion molecules, and tissue development-related molecules and down-regulated the expression of chemokines (CXCL10 and CXCL8) and adaptive immune response-related molecules. GF(IL-17) up-regulated the expression of chemokines and other molecules for neutrophil infiltration and activation, the pro-inflammatory cytokine IL-6, and C3. GF(TGF-β) up-regulated the expression of cell growth-related molecules, ECM components, several types of collagen, and cell adhesion molecules and down-regulated the expression of molecules related to complement activation and bacterial recognition. GFs stimulated by 4 cytokines responded differently to LPS. These results show that Th cytokines can polarize GFs into cells with functionally distinct features: immune-activating but tissue-destructive GF(IFN-γ), tissue-reparative, and immune-inhibiting GF(IL-4), highly pro-inflammatory GF(IL-17), and potent tissue-reparative GF(TGF-β).

Adipogenesis or osteogenesis: destiny decision made by mechanical properties of biomaterials.

Regenerative medicine affords an effective approach for restoring defect-associated diseases, and biomaterials play a pivotal role as cell niches to support the cell behavior and decide the destiny of cell differentiation. Except for chemical inducers, mechanical properties such as stiffness, pore size and topography of biomaterials play a crucial role in the regulation of cell behaviors and functions. Stiffness may determine the adipogenesis or osteogenesis of mesenchymal stem cells (MSCs) via the translocation of yes-associated protein (YAP) and the transcriptional coactivator with a PDZ-binding motif (TAZ). External forces transmit through cytoskeleton reorientation to assist nuclear deformation and molecule transport, meanwhile, signal pathways including the Hippo, FAK/RhoA/ROCK, and Wnt/β-catenin have been evidenced to participate in the mechanotransduction. Different pore sizes not only tailor the scaffold stiffness but also conform to the requirements of cell migration and vessels in-growth. Topography guides cell geometry along with mobility and determines the cell fate ascribed to micro/nano-scale contact. Herein, we highlight the recent progress in exploring the regulation mechanism by the physical properties of biomaterials, which might lead to more innovative regenerative strategies for adipose or bone tissue repair.

Histone H2A Nuclear/Cytoplasmic Trafficking Is Essential for Negative Regulation of Antiviral Immune Response and Lysosomal Degradation of TBK1 and IRF3

Histone H2A is a nuclear molecule tightly associated in the form of the nucleosome. Our previous studies have demonstrated the antibacterial property of piscine H2A variants against gram-negative bacteria Edwardsiella piscicida and Gram-positive bacteria Streptococcus agalactiae. In this study, we show the function and mechanism of piscine H2A in the negative regulation of RLR signaling pathway and host innate immune response against spring viremia of carp virus (SVCV) infection. SVCV infection significantly inhibits the expression of histone H2A during an early stage of infection, but induces the expression of histone H2A during the late stage of infection such as at 48 and 72 hpi. Under normal physiological conditions, histone H2A is nuclear-localized. However, SVCV infection promotes the migration of histone H2A from the nucleus to the cytoplasm. The in vivo studies revealed that histone H2A overexpression led to the increased expression of SVCV gene and decreased survival rate. The overexpression of histone H2A also significantly impaired the expression levels of those genes involved in RLR antiviral signaling pathway. Furthermore, histone H2A targeted TBK1 and IRF3 to promote their protein degradation via the lysosomal pathway and impair the formation of TBK1-IRF3 functional complex. Importantly, histone H2A completely abolished TBK1-mediated antiviral activity and enormously impaired the protein expression of IRF3, especially nuclear IRF3. Further analysis demonstrated that the inhibition of histone H2A nuclear/cytoplasmic trafficking could relieve the protein degradation of TBK1 and IRF3, and blocked the negative regulation of histone H2A on the SVCV infection. Collectively, our results suggest that histone H2A nuclear/cytoplasmic trafficking is essential for negative regulation of RLR signaling pathway and antiviral immune response in response to SVCV infection.

A two state model study of photo‐detachment dynamics driven by an optimally designed polychromatic field: A simulated annealing based optimisation

AbstractThe surfaces of and F2 are two well separated surfaces modeled by two Morse potentials. In presence of a field, these two surfaces couple and transition from to F2 takes place resulting in the detachment of an electron. We have applied Simulated Annealing (SA) technique to design the polychromatic field which results in ∼ 95% probability of photo detachment (Pd). Also we have designed a field to achieve photo detachment followed by dissociation of FF bond with nearly 38% dissociation probability. F2 being a homo nuclear diatomic molecule, promotion to the continuum states by directly exciting it from its ground vibrational state is an impossibility and our method appears to be a feasible solution if the target is to break the FF bond.

SYVN1-MTR4-MAT2A Signaling Axis Regulates Methionine Metabolism in Glioma Cells.

Methionine is one of the essential amino acids. How tumor cells adapt and adjust their signal transduction networks to avoid apoptosis in a methionine-restricted environment is worthy of further exploration. In this study, we investigated the molecular mechanism of glioma response to methionine restriction, providing a theoretical basis for new treatment strategies for glioma.MethodsWe constructed methionine-restriction-tolerant cells in order to study the response of glioma to a methionine-restricted environment. The transcriptome analysis of the tolerant cells showed significant changes in MAT2A. Western blotting, immunohistochemistry, quantitative real-time PCR, colony formation assays, and other experiments were used to verify the role of MAT2A in glioma genesis. In addition, the regulatory mechanism of MAT2A mRNA nuclear export was investigated by transfection, plasma nucleation separation, and co-immunoprecipitation.ResultsUnder methionine restriction, glioma cells showed high expression of MAT2A, and an inhibitor of MAT2A reduced the proliferation of tumor cells. The expression of MAT2A was positively correlated with World Health Organization-grade glioma. High expression of MAT2A was related to increased transfer of its mRNA out of the nucleus. The expression of nuclear export regulatory molecule MTR4 could affect the export of MAT2A mRNA. In a methionine-restricted environment, ubiquitination of MTR4 was enhanced, and thus its protein level was reduced. The E3 ubiquitin ligase was verified to be SYVN1.ConclusionIn summary, methionine restriction leads to increased ubiquitination of MTR4, which promotes the transfer of MAT2A mRNA out of the nucleus and MAT2A protein expression. MAT2A promotes histone methylation, prompting cells to proliferate in a methionine-restricted environment.