Extracellular Medium Research Articles

The rostromedial tegmental nucleus RMTg is not a critical site for ethanol-induced motor activation in rats.

Opioid drugs indirectly activate dopamine (DA) neurons in the ventral tegmental area (VTA) through a disinhibition mechanism mediated by mu opioid receptors (MORs) present both on the GABA projection neurons located in the medial tegmental nucleus/tail of the VTA (RMTg/tVTA) and on the VTA GABA interneurons. It is well demonstrated that ethanol, like opioid drugs, provokes VTA DA neuron disinhibition by interacting (through its secondary metabolite, salsolinol) with MORs present in VTA GABA interneurons, but it is not known whether ethanol could disinhibit VTA DA neurons through the MORs present in the RMTg/tVTA. The objective of the present study was to determine whether ethanol, directly microinjected into the tVTA/RMTg, is also able to induce VTA DA neurons disinhibition. Disinhibition of VTA DA neurons was indirectly assessed through the analysis of the motor activity of rats. Cannulae were placed into the tVTA/RMTg to perform microinjections of DAMGO (0.13 nmol), ethanol (150 or 300 nmol) or acetaldehyde (250 nmol) in animals pre-treated with either aCSF or the irreversible antagonist of MORs, beta-funaltrexamine (beta-FNA; 2.5 nmol). After injections, spontaneous activity was monitored for 30 min. Neither ethanol nor acetaldehyde directly administered into the RMTg/tVTA were able to increase the locomotor activity of rats at doses that, in previous studies performed in the posterior VTA, were effective in increasing motor activities. However, microinjections of 0.13 nmol of DAMGO into the tVTA/RMTg significantly increased the locomotor activity of rats. These activating effects were reduced by local pre-treatment of rats with beta-FNA (2.5 nmol). The tVTA/RMTg does not appear to be a key brain region for the disinhibiting action of ethanol on VTA DA neurons. The absence of dopamine in the tVTA/RMTg extracellular medium, the lack of local ethanol metabolism or both could explain the present results.

Acidosis-induced regulation of adipocyte G0S2 promotes crosstalk between adipocytes and breast cancer cells as well as tumor progression

Bidirectional interactions between cancer cells and their microenvironment govern tumor progression. Among the stromal cells in this microenvironment, adipocytes have been reported to upregulate cancer cell migration and invasion by producing fatty acids. Conversely, cancer cells alter adipocyte phenotype notably via increased lipolysis. We aimed to identify the mechanisms through which cancer cells trigger adipocyte lipolysis and evaluate the functional consequences on cancer progression. Here, we show that cancer cell-induced acidification of the extracellular medium strongly promotes preadipocyte lipolysis through a mechanism that does not involve lipophagy but requires adipose triglyceride lipase (ATGL) activity. This increased lipolysis is triggered mainly by attenuation of the G0/G1 switch gene 2 (G0S2)-induced inhibition of ATGL. G0S2-mediated regulation in preadipocytes affects their communication with breast cancer cells, modifying the phenotype of the cancer cells and increasing their resistance to chemotherapeutic agents in vitro. Furthermore, we demonstrate that the adipocyte-specific overexpression of G0S2 impairs mammary tumor growth and lung metastasis formation in vivo. Our results highlight the importance of acidosis in cancer cell-adipocyte crosstalk and identify G0S2 as the main regulator of cancer-induced lipolysis, regulating tumor establishment and spreading.

Mesenchymal Stem Cells: Their Role in the Tumor Microenvironment.

Mesenchymal stem cells (MSCs) have been seen for years as great candidates for treating different diseases and an alternative to embryonic stem cells due to their differentiation capacity in vitro. More recent research has focused on their ability to modulate the immune response and regeneration at sites associated with inflammation, activities attributable to the release of trophic factors into the extracellular medium, a set of components known as the secretome. It has been possible to demonstrate the presence of these cells within the tumor microenvironment, which is associated with their tropism for sites of inflammation; however, their role here needs to be clarified. In different investigations, the feasibility of using MSCs or their secretome to treat cancer has been sought, with these results being ambiguous. It has been described that MSCs can be activated and present various phenotypes, which could explain the divergence in their action; however, these activation mechanisms and the different phenotypes still need to be well known. This review explores MSCs and their use in regenerative medicine with a targeted approach to cancer. Impact Statement This text addresses the diverging findings on the role of mesenchymal stem cells in the tumor microenvironment and discrepancies on the use of these cells as cancer treatment, separating the direct use of the cells from the use of the secretome. Multiple authors refer equally to the cells and their secretome to conclude on the positive or negative outcome, without taking into consideration how the cells are affected by their surroundings.

Spheresomes are the main extracellular vesicles in low-grade gliomas

Cancer progression and its impact on treatment response and prognosis is deeply regulated by tumour microenvironment (TME). Cancer cells are in constant communication and modulate TME through several mechanisms, including transfer of tumour-promoting cargos through extracellular vesicles (EVs) or oncogenic signal detection by primary cilia. Spheresomes are a specific EV that arise from rough endoplasmic reticulum–Golgi vesicles. They accumulate beneath cell membrane and are released to the extracellular medium through multivesicular spheres. This study describes spheresomes in low-grade gliomas using electron microscopy. We found that spheresomes are more frequent than exosomes in these tumours and can cross the blood–brain barrier. Moreover, the distinct biogenesis processes of these EVs result in unique cargo profiles, suggesting different functional roles. We also identified primary cilia in these tumours. These findings collectively contribute to our understanding of glioma progression and metastasis.

A comparison of SNARF-1 and skeletal δ11B estimates of calcification media pH in tropical coral

Coral skeletal boron geochemistry offers opportunities to probe the pH of the calcification media (pHCM) of modern and fossil specimens, to estimate past changes in seawater pH and to explore the biomineralisation response to future ocean acidification. In this research we grew 2 Stylophora pistillata coral microcolonies over glass coverslips to allow analysis of the pH sensitive dye SNARF-1, in the extracellular calcification medium at the growing edge of colonies where the first aragonite crystals are formed, under both light and dark conditions. We use secondary ion mass spectrometry (SIMS) to measure the boron isotopic composition (δ11B) of the skeleton close to the growth edge after 2 to 3 days of additional calcification had enlarged the crystals until they joined, generating a continuous sheet of aragonite. Mean skeletal δ11B-pHCM estimates are higher than those by SNARF-1 by 0.35–0.44 pH units. These differences either reflect real variations in the pH of the calcification media associated with each measurement technique or indicate other changes in the biomineralisation process which influence skeletal δ11B. SNARF-1 measures directly the pH of the extracellular calcification medium while skeletal δ11B analyses aragonite potentially formed via both extracellular and intracellular biomineralisation pathways. Analysis of a third coral specimen, also growing on a glass slide but with a 5 cm long branch, indicated good agreement between the δ11B value of the apex of the branch and the skeletal growth edge. The tissues overlying both these regions were transparent indicating they had low symbiont densities. This suggests that the biomineralisation process is broadly comparable between these sites and that studies growing corals over glass slides/coverslips provide representative data for the colony apex.

Time Kill Kinetic, Antibiofilm Activity and Effect on Biomolecules Release of Drypetes Gosswelleri and Echinops Giganteus Essential Oils on Bacteria

Finding alternative strategies to confront bacterial resistance is an urgent need. Biofilm-forming bacteria have become a serious problem in medicine. Bacteria can use biofilm as a mechanism of resistance against antibacterial drugs. The aim of this study was to study times kill kinetic of Drypetes gosswelleri and Echinops giganteus essential oils, the antibiofilm formation activity and their effect on cell release compounds. Times kill kinetic was study by quantification of cellular growth over time in Petri dish after her inoculation by the cells treated with antibacteria at MBC and incubation at different times during 24h. The antibiofilm activity carried out by microdilution using 96 wells microplate. The production of biofilms by cells treated was observed and quantified after coloration by Crystal Violet dye and spectrophotometric dosage at 630nm. The effect on cell release compounds are study by determination of absorbent material in extracellular medium at 260nm after exposure at the antibacterial during 24h. The results obtained showed that, these EOs have an ability to kill bacterial cells over time, Drypetes gossweileri EOs like Ciprofloxacin, caused the death of all the cells in the inoculum treated before 14h of exposure of Staphylococcus aureus strain. Against Salmonella enteritidis strain, Drypetes gossweileri and Echnops giganteus EOs kill all the cells after 24h like the two antibiotics. One effect of the action these EOs on the strains elucidated in this study was the leakage of intracellular absorbent materials (DNA and RNA) this materialized a diminution of membrane permeability and cell wall integrity.

EVALUATION OF ETHANOL AND BIOMASS PRODUCTION RATE BY DIFFERENT $SACCHAROMYCES~CEREVISIAE$ STRAINS DEPENDING ON EXTERNAL pH AND TEMPERATURE

Bioethanol production is in high demand due to its potential to replace transportation fuel and its ability to be renewable and long-lasting. Therefore, the bioconversion of fermentable sugars to ethanol is of central importance. The work aimed to explore yeast properties changes during fermentation at different pHs and temperature conditions. Biomass production, specific growth rate (SGR), fermentation products, and metabolite composition and quantity changes were studied at 25℃, 30℃, and 37℃ as well as pH 5 and pH 6.5 conditions. SGR data shows that at 25℃ and 30℃ pH 5 is optimal for yeast biomass production. At pH 5.0–6.5, the biomass production of the ATCC 9804 strain almost 2 times exceeds the same parameter for another strain. The highest biomass production was detected at 30℃ and pH 5. Ethanol production by yeast increases as the growth temperature decreases at pH 5 in contrast to pH 6.5, where the temperature changes within 30–37℃ range have no significant effect on it in both strains. At 30℃ ethanol concent­ra­tion in the extracellular medium reached ~117 mM in the case of ATCC 13007 strain, whereas for ATCC 9804 the same parameter was 1.4-fold lower. Both strains had the same substrate assimilation rate. Glycerol production reduces with increasing growth temperature and pH; the highest glycerol concentration (6.1 mM) was observed within 32 h growth of ATCC 9804 strain at 25℃, pH 5 and reaches 5.5 mM within 24 h growth of ATCC 13007 strain under the same conditions. At pH 5 and 6.5, the free energy for glycerol production was 2.3-fold and 4-fold higher compared to that of ethanol and acetate production processes in both strains. Gibbs free energy of ethanol production reaches the lowest value compared to the same parameter of acetate and glycerol production at pH 5, suggesting a preference for the alcoholic fermentation metabolic pathway under these conditions. The lowest acetate production was observed after 24 hours of growth of the ATCC 13007 strain at 37°C and pH 5, with ΔG = 173.9 kJ/mol. Obtained data highlights temperature stress mechanisms regulation of yeast and can be used for improving ethanol production processes and yeast genetic modification tools.

Modeling collagen fibril self-assembly from extracellular medium in embryonic tendon

Collagen is a key structural component of multicellular organisms and is arranged in a highly organized manner. In structural tissues such as tendons, collagen forms bundles of parallel fibers between cells, which appear within a 24-h window between embryonic day 13.5 (E13.5) and E14.5 during mouse embryonic development. Current models assume that the organized structure of collagen requires direct cellular control, whereby cells actively lay down collagen fibrils from cell surfaces. However, such models appear incompatible with the time and length scales of fibril formation. We propose a phase-transition model to account for the rapid development of ordered fibrils in embryonic tendon, reducing reliance on active cellular processes. We develop phase-field crystal simulations of collagen fibrillogenesis in domains derived from electron micrographs of inter-cellular spaces in embryonic tendon and compare results qualitatively and quantitatively to observed patterns of fibril formation. To test the prediction of this phase-transition model that free protomeric collagen should exist in the inter-cellular spaces before the formation of observable fibrils, we use laser-capture microdissection, coupled with mass spectrometry, which demonstrates steadily increasing free collagen in inter-cellular spaces up to E13.5, followed by a rapid reduction of free collagen that coincides with the appearance of less-soluble collagen fibrils. The model and measurements together provide evidence for extracellular self-assembly of collagen fibrils in embryonic mouse tendon, supporting an additional mechanism for rapid collagen fibril formation during embryonic development.

Real-time monitoring of cell surface protein arrival with split luciferases.

Each cell in a multicellular organism permanently adjusts the concentration of its cell surface proteins. In particular, epithelial cells tightly control the number of carriers, transporters and cell adhesion proteins at their plasma membrane. However, sensitively measuring the cell surface concentration of a particular protein of interest in live cells and in real time represents a considerable challenge. Here, we introduce a novel approach based on split luciferases, which uses one luciferase fragment as a tag on the protein of interest and the second fragment as a supplement to the extracellular medium. Once the protein of interest arrives at the cell surface, the luciferase fragments complement and generate luminescence. We compared the performance of split Gaussia luciferase and split Nanoluciferase by using a system to synchronize biosynthetic trafficking with conditional aggregation domains. The best results were achieved with split Nanoluciferase, for which luminescence increased more than 6000-fold upon recombination. Furthermore, we showed that our approach can separately detect and quantify the arrival of membrane proteins at the apical and basolateral plasma membrane in single polarized epithelial cells by detecting the luminescence signals with a microscope, thus opening novel avenues for characterizing the variations in trafficking in individual epithelial cells.

O-282 Role of extra-cellular vesicles in reproduction

Abstract Extracellular vesicles (EVs) are bilayer-membrane-bound vesicles isolated from biofluids and tissues. Recently, these organelles have been recognized as a novel mode for inter and intra-cellular communication. The three critical subtypes of EVs include exosomes, microvesicles, and apoptotic bodies. They can be discriminated according to their size, cargo, biogenesis, release pathways, and functions. These organelles serve as vehicles for transferring bioactive molecules as lipids, proteins, cytokines, and regulatory molecules as RNAs, locally and remotely. A wide range of cell types releases EVs, and their cargo differs under normal and pathological conditions. These facts make them possible biomarkers for diagnosing and treating different medical conditions. Isolation of EVs from various female and male reproductive tissues such as prostate, epididymis, seminal fluid, fallopian tube, cumulus, and mural granulosa cells, oocytes, embryos at various developmental stages, endometrium, and decidua has been described. Oogenesis, sperm maturation, fertilization, embryo development, and implantation are composite processes highly reliant on the interaction between tissues and cells. Existing data show that miRNAs and proteins in epididymal fluid associated with testicular maturation are transferred to the sperm by EVs Ovarian follicular growth is a synchronized process comprising bidirectional communication between the oocyte, cumulus, and mural granulosa cells. Some studies have identified EVs from several species' follicular fluid (FF), including bovine, equine, and human. To date, data linking EVs with oogenesis are primarily descriptive. EV micro-RNA (EV-miRNA) isolated from FF target pathways as mitogen-activated protein kinase (MAPK), wingless signaling pathway (WNT), epidermal growth factor receptor (ErbB), and transforming growth factor beta (TGFß). These pathways are related to folliculogenesis, meiotic resumption, and ovulation. EV-miRNAs isolated from follicles that contained mature oocytes were associated with fertilization potential and embryo quality. In addition, EVs derived from microvillous-rich oolemma may neutralize acrosome reaction and prevent polyspermy. Animal and human studies have confirmed that embryos at various developmental stages secrete EVs into the extracellular medium, and embryos internalize EVs. The number, miRNA profile, and size of EVs secreted vary across the cycle and according to embryos’ developmental potential. Endometrial EVs release miRNAs essential for the blastocyst's adhesion. In ovine, EVs were identified from the uterine lumen and uterine flushing during implantation. Uterine lumen flush EVs were internalized by the trophectoderm cells in the conceptus, reducing trophectoderm cell proliferation. The study of EVs in reproduction has increased exponentially in the last few years. Evidence support that these organelles contribute to oogenesis, fertilization, embryo development, and implantation. Further research in EVs will help us expand noninvasive methods and identify new biomarkers in the reproductive field.

Exploring the involvement of TASK-1 in the control of isolated rat right atrium function from healthy animals and an experimental model of monocrotaline-induced pulmonary hypertension.

The TASK-1 channel belongs to the two-pore domain potassium channel family. It is expressed in several cells of the heart, including the right atrial (RA) cardiomyocytes and the sinus node, and TASK-1 channel has been implicated in the pathogenesis of atrial arrhythmias (AA). Thus, using the rat model of monocrotaline-induced pulmonary hypertension (MCT-PH), we explored the involvement of TASK-1 in AA. Four-week-old male Wistar rats were injected with 50 mg/kg of MCT to induce MCT-PH and isolated RA function was studied 14 days later. Additionally, isolated RA from six-week-old male Wistar rats were used to explore the ability of ML365, a selective blocker of TASK-1, to modulate RA function. The hearts developed right atrial and ventricular hypertrophy, inflammatory infiltrate and the surface ECG demonstrated increased P wave duration and QT interval, which are markers of MCT-PH. The isolated RA from the MCT animals showed enhanced chronotropism, faster contraction and relaxation kinetics, and a higher sensibility to extracellular acidification. However, the addition of ML365 to extracellular media was not able to restore the phenotype. Using a burst pacing protocol, the RA from MCT animals were more susceptible to develop AA, and simultaneous administration of carbachol and ML365 enhanced AA, suggesting the involvement of TASK-1 in AA induced by MCT. TASK-1 does not play a key role in the chronotropism and inotropism of healthy and diseased RA; however, it may play a role in AA in the MCT-PH model.

Glucose Transport in Escherichia coli: From Basics to Transport Engineering.

Escherichia coli is the best-known model for the biotechnological production of many biotechnological products, including housekeeping and heterologous primary and secondary metabolites and recombinant proteins, and is an efficient biofactory model to produce biofuels to nanomaterials. Glucose is the primary substrate used as the carbon source for laboratory and industrial cultivation of E. coli for production purposes. Efficient growth and associated production and yield of desired products depend on the efficient sugar transport capabilities, sugar catabolism through the central carbon catabolism, and the efficient carbon flux through specific biosynthetic pathways. The genome of E. coli MG1655 is 4,641,642 bp, corresponding to 4702 genes encoding 4328 proteins. The EcoCyc database describes 532 transport reactions, 480 transporters, and 97 proteins involved in sugar transport. Nevertheless, due to the high number of sugar transporters, E. coli uses preferentially few systems to grow in glucose as the sole carbon source. E. coli nonspecifically transports glucose from the extracellular medium into the periplasmic space through the outer membrane porins. Once in periplasmic space, glucose is transported into the cytoplasm by several systems, including the phosphoenolpyruvate-dependent phosphotransferase system (PTS), the ATP-dependent cassette (ABC) transporters, and the major facilitator (MFS) superfamily proton symporters. In this contribution, we review the structures and mechanisms of the E. coli central glucose transport systems, including the regulatory circuits recruiting the specific use of these transport systems under specific growing conditions. Finally, we describe several successful examples of transport engineering, including introducing heterologous and non-sugar transport systems for producing several valuable metabolites.

Identification and molecular characterization of two recurrent missense mutations in the RS1 gene in two families with X-linked retinoschisis from North India.

X-linked retinoschisis (XLR) is a rare medical condition that involves in the splitting of neurosensory layers and the impairment of vision in the retina. In majority of the XLR cases, pathogenic variants in Retinoschisin 1 (RS1) gene have been implicated in males with an early age of onset during early childhood. In the present study, we have recruited two North Indian families having multiple affected male members, who were diagnosed with XLR. The entire protein-coding region of RS1 was screened by PCR-Sanger sequencing and two recurrent pathogenic variants (p.I81N and p.R102Q) were unraveled. The in vitro study of these variants demonstrated the aggregation of mutant RS1 within the endoplasmic reticulum. Furthermore, mutant forms of this protein showed significant intracellular retention, which was evident by the absence of retinoschisin protein fractions in the extracellular media. These inferences were also supported by extensive bioinformatics analysis of the mutants, which showed dramatic conformational changes in the local structure of retinoschisin. Thus, our study suggests that the identified pathogenic variants interfere with proper protein folding, leading to anomalous structural changes ultimately resulting in intracellular retention of retinoschisin within the retina.

Single-cell ICP-MS for studying the association of inorganic nanoparticles with cell lines derived from aquaculture species

The current research deals with the use of single-cell inductively coupled plasma-mass spectrometry (scICP-MS) for the assessment of titanium dioxide nanoparticle (TiO2 NP) and silver nanoparticle (Ag NP) associations in cell lines derived from aquaculture species (sea bass, sea bream, and clams). The optimization studies have considered the avoidance of high dissolved background, multi-cell peak coincidence, and possible spectral interferences. Optimum operating conditions were found when using a dwell time of 50 μs for silver and 100 μs for titanium. The assessment of associated TiO2 NPs by scICP-MS required the use of ammonia as a reaction gas (flow rate at 0.75 mL min−1) for interference-free titanium determinations (measurements at an m/z ratio of 131 from the 48Ti(NH)(NH3)4 adduct). The influence of other parameters such as the number of washing cycles and the cell concentration on accurate determinations by scICP-MS was also fully investigated. Cell exposure trials were performed using PVP-Ag NPs (15 and 100 nm, nominal diameter) and citrate-TiO2 NPs (5, 25, and 45 nm, nominal diameter) at nominal concentrations of 10 and 50 μg mL−1 for citrate-TiO2 NPs and 5.0 and 50 μg mL−1 for PVP-Ag NPs. Results have shown that citrate-TiO2 NPs interact with the outer cell membranes, being quite low in the number of citrate-TiO2 NPs that enters the cells (the high degree of aggregation is the main factor which leads to the aggregates being in the extracellular medium). In contrast, PVP-Ag NPs have been found to enter the cells.Graphical abstract

New insights into signal transduction pathways in adrenal steroidogenesis: role of mitochondrial fusion, lipid mediators, and MAPK phosphatases.

Hormone-receptor signal transduction has been extensively studied in adrenal gland. Zona glomerulosa and fasciculata cells are responsible for glucocorticoid and mineralocorticoid synthesis by adrenocorticotropin (ACTH) and angiotensin II (Ang II) stimulation, respectively. Since the rate-limiting step in steroidogenesis occurs in the mitochondria, these organelles are key players in the process. The maintenance of functional mitochondria depends on mitochondrial dynamics, which involves at least two opposite events, i.e., mitochondrial fusion and fission. This review presents state-of-the-art data on the role of mitochondrial fusion proteins, such as mitofusin 2 (Mfn2) and optic atrophy 1 (OPA1), in Ang II-stimulated steroidogenesis in adrenocortical cells. Both proteins are upregulated by Ang II, and Mfn2 is strictly necessary for adrenal steroid synthesis. The signaling cascades of steroidogenic hormones involve an increase in several lipidic metabolites such as arachidonic acid (AA). In turn, AA metabolization renders several eicosanoids released to the extracellular medium able to bind membrane receptors. This report discusses OXER1, an oxoeicosanoid receptor which has recently arisen as a novel participant in adrenocortical hormone-stimulated steroidogenesis through its activation by AA-derived 5-oxo-ETE. This work also intends to broaden knowledge of phospho/dephosphorylation relevance in adrenocortical cells, particularly MAP kinase phosphatases (MKPs) role in steroidogenesis. At least three MKPs participate in steroid production and processes such as the cellular cycle, either directly or by means of MAP kinase regulation. To sum up, this review discusses the emerging role of mitochondrial fusion proteins, OXER1 and MKPs in the regulation of steroid synthesis in adrenal cortex cells.

DNA methyltransferases inhibitor azacitidine improves the skeletal phenotype of mild osteogenesis imperfecta by reversing the impaired osteogenesis and excessive osteoclastogenesis.

Osteogenesis imperfecta (OI), as a disease of congenital bone dysplasia, is often accompanied by the abnormal alteration of bone absorption and bone formation. DNA methyltransferases (Dnmts) can regulate the gene expression involved in osteogenesis and osteoclastogenesis. Dnmts changes and their effects on bone cells under OI is poorly understood. The Dnmts expression in adipose derived mesenchymal stem cells (ADSCs), bone marrow derived pre-osteoclasts (pre-Ocs) and femurs of Col1a2oim/+ and Col1a1+/-365 mice, both modeling mild OI types, were determined. The effects of azacitidine (Aza) administration and Dnmt3a knockdown by ShRNA on the osteogenic differentiation of ADSCs together with osteoclasts (Ocs) production of pre-Ocs were studied in vitro. The synthesis and secretion of collagen fibers of OI derived ADSCs were examined. The therapeutic outcomes of intraperitoneal (i.p.) infused Aza (1mg/kg/2d) for 30days were evaluated in OI mice. Obviously elevated expression of Dnmts, especially Dnmt3a, existed in ADSCs, pre-Ocs, and femurs isolated from OI modeled mice. Much more collagen molecules of mutant ADSCs were secreted into the extracellular medium post Aza addition. Both Aza administration and Dnmt3a knockdown effectively enhanced the bone-forming capacity of affected ADSCs and reduced Ocs formation of OI mice in vitro. Aza treatment apparently improved the femora microstructure and biomechanical properties, increased bone formation and decreased the number of Ocs in mice with OI. Highly expressed Dnmt3a contributed to the impaired osteogenesis and enhanced osteoclastogenesis of collagen defect-related OI. Aza medication effectively improved the femora phenotype of the two types of OI modeled mice partly by Dnmts inhibition and modulating cell stress response. These findings facilitated understanding the role of Dnmts alteration in skeletal pathological development of mild OI and preliminary confirmed the therapeutic potential of Dnmts depressants in mild OI treatment. Still, further researches are needed to explore the specific function of Dnmts in OI bones and clarify the benefits of Aza administration in OI treatment.

Bursting Dynamics of Spiking Neural Network Induced by Active Extracellular Medium

We propose a mathematical model of a spiking neural network (SNN) that interacts with an active extracellular field formed by the brain extracellular matrix (ECM). The SNN exhibits irregular spiking dynamics induced by a constant noise drive. Following neurobiological facts, neuronal firing leads to the production of the ECM that occupies the extracellular space. In turn, active components of the ECM can modulate neuronal signaling and synaptic transmission, for example, through the effect of so-called synaptic scaling. By simulating the model, we discovered that the ECM-mediated regulation of neuronal activity promotes spike grouping into quasi-synchronous population discharges called population bursts. We investigated how model parameters, particularly the strengths of ECM influence on synaptic transmission, may facilitate SNN bursting and increase the degree of neuronal population synchrony.

Geochemical responses of scleractinian corals to nutrient stress

The impact of changing nutrient conditions on scleractinian coral-based geochemical proxies is poorly understood, despite the nutrient balance in many coral reefs being disturbed by anthropogenic activity. Here, the geochemical responses of tropical corals Acropora polystoma and Porites lichen to nutrient enrichment and depletion are examined following growth under cultured conditions, to assess the impact of nutrients on traditional geochemical proxies for both temperature and the coral internal carbonate system. The corals were exposed to four different nutrient treatments over a period of 140 days: (1) a replete treatment with optimal levels of nitrate (∼4.5 μM) and phosphate (∼0.6 μM), (2) a nutrient depleted treatment with negligible nitrate and phosphate, (3) a treatment with high nitrate (∼73 μM) and negligible phosphate, and (4) a treatment with high phosphate (∼5.7 μM) and negligible nitrate. Results suggest nutrients play a hitherto under-appreciated role in coral skeleton elemental (Li/Ca, B/Ca, Mg/Ca, Sr/Ca, Li/Mg) and isotopic (δ11B) composition, with the internal carbonate chemistry also impacted. For example, Mg/Ca and Sr/Ca are lower, and Li/Mg higher, in the nutrient imbalanced and deplete treatments compared to the replete treatment for both species. Disruption to the carbonate system in corals cultured under imbalanced nutrient conditions is best explained by a decrease in dissolved inorganic carbon flux to the extracellular calcifying medium. Variations in nutrient concentration — or nutrient imbalance — can have dramatic consequences on both reconstructed sea surface temperatures and ocean or calcification pH, with reconstructed temperatures varying from −7°C to +52 °C, and δ11B-derived pH by up to 0.13 pH units. The impact from anthropogenically-induced nutrient disturbances should therefore be considered when generating temporal records of environments using coral skeletal archives.

Impact of interorganelle coordination between the conventional early secretory pathway and autophagy in cellular homeostasis and stress response.

The conventional early secretory pathway and autophagy are two essential interconnected cellular processes that are crucial for maintaining cellular homeostasis. The conventional secretory pathway is an anabolic cellular process synthesizing and delivering proteins to distinct locations, including different organelles, the plasma membrane, and the extracellular media. On the other hand, autophagy is a catabolic cellular process that engulfs damaged organelles and aberrant cytosolic constituents into the double autophagosome membrane. After fusion with the lysosome and autolysosome formation, this process triggers digestion and recycling. A growing list of evidence indicates that these anabolic and catabolic processes are mutually regulated. While knowledge about the molecular actors involved in the coordination and functional cooperation between these two processes has increased over time, the mechanisms are still poorly understood. This review article summarized and discussed the most relevant evidence about the key molecular players implicated in the interorganelle crosstalk between the early secretory pathway and autophagy under normal and stressful conditions.

Targeting G-quadruplex motifs interferes with differentiation of adipose-derived mesenchymal stem cells

BackgroundG-quadruplex (G4) motifs are nucleic acid secondary structures observed in mammalian genomes and transcriptomes able to regulate various cellular processes. Several small molecules have been developed so far to modulate G4 stability, frequently associated with anticancer activity. However, how G4 structures are regulated over homeostatic conditions is mostly unexplored. Here, we used human adipose-derived mesenchymal stem cells (ASCs) to address the role of G4 motifs during adipogenic differentiation.MethodsAdipocyte differentiation of ASCs was investigated in the presence or absence of a well-known G4 ligand, Braco-19. Cell viability was determined by sulforhodamine B assay. Cell dimension and granularity, DNA G4 motifs and cell cycle were detected by flow cytometry. Lipid droplet accumulation was assessed by Oil Red O staining. Cell senescence was evaluated by β-galactosidase staining. Gene expression was measured by qPCR. Protein release in the extracellular medium was quantified by ELISA.ResultsBraco-19 used at non-cytotoxic concentrations induced morphological changes in mature adipocytes partially restoring an undifferentiated-like status. Braco-19 reduced lipid vacuolization and PPARG, AP2, LEP and TNFA mRNA levels in terminally differentiated cells. No effect was observed in cell senescence, fibrotic markers, IL-6 and IL-8 production, while the secretion of VEGF was dose-dependently reduced. Interestingly, G4 structures were increased in differentiated adipocytes compared to their precursors. Braco-19 treatment reduced G4 content in mature adipocytes.ConclusionsOur data highlight a new role of G4 motifs as genomic structural elements related to human ASC differentiation into mature adipocytes, with potential implications in physio-pathological processes.