Changes In Cell Size Research Articles

Green Synthesis of Organo-Montmorillonite/Silver Nanocomposites on Dyed Cotton with Vat Dyes to Achieve Biocompatible Antibacterial Properties on Fashionable Clothing

ABSTRACT This study proposes a green and easy route for synthesizing clay/silver nanocomposite on denim fabric which can instantaneously discolor the garment and produce antibacterial effects during a one-step process with minimal toxicity to humans. Synthesis of organo-Montmorillonite/silver nanocomposite was carried out by reducing silver nitrate to nanosilver in the presence of a reducing agent and its attachment to opened layers of clay particles throughout a one-step process. Colorimetric properties were examined using L*a*b* values to understand the changes in denim color after the action of organo-montmorillonite (O-MMt) and nanosilver, which confirmed the dye removal action of clay/silver nanocomposite. UV-visible data proved the successful synthesis of nanosilver in addition to the indigo dye removal from the remaining effluent by exhibiting no peaks at 406 nm and 601.2 nm. SEM and TEM images depicted the shape and position of opened O-MMt layers, silver nanoparticles, and O-MMt/silver nanocomposites, verified by EDX, XRD, and FTIR data. Antibacterial examination showed noticeable results with maximum effectiveness of 100% against the growth of Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus bacteria with acceptable cell viability of 94% with no detectable changes in cell shape and size.

Molecular Dynamics Simulations of the Breathing Phase Transition of MOF Nanocrystallites II: Explicitly Modeling the Pressure Medium.

One of the most investigated properties of porous crystalline metal-organic frameworks (MOFs) is their potential flexibility to undergo large changes in unit cell size upon guest adsorption or other stimuli, referred to as “breathing”. Computationally, such phase transitions are usually investigated using periodic boundary conditions, where the system’s volume can be controlled directly. However, we have recently shown that important aspects like the formation of a moving interface between the open and the closed pore form or the free energy barrier of the first-order phase transition and its size effects can best be investigated using non-periodic nanocrystallite (NC) models [Keupp et al. (Adv. Theory Simul., 2019, 2, 1900117)]. In this case, the application of pressure is not straightforward, and a distance constraint was used to mimic a mechanical strain enforcing the reaction coordinate. In contrast to this prior work, a mediating particle bath is used here to exert an isotropic hydrostatic pressure on the MOF nanocrystallites. The approach is inspired by the mercury nanoporosimetry used to compress flexible MOF powders. For such a mediating medium, parameters are presented that require a reasonable additional numerical effort and avoid unwanted diffusion of bath particles into the MOF pores. As a proof-of-concept, NCs of pillared-layer MOFs with different linkers and sizes are studied concerning their response to external pressure exerted by the bath. By this approach, an isotropic pressure on the NC can be applied in analogy to corresponding periodic simulations, without any bias for a specific mechanism. This allows a more realistic investigation of the breathing phase transformation of a MOF NC and further bridges the gap between experiment and simulation.

Effect of Lowered pH on Community Composition, Growth and Cell Morphology of Freshwater Phytoplankton

This experiment studied the probable effects of lowered pH (6.5 to 5.0) on four freshwater phytoplankton species Chlorella vulgaris Beyerinck, Scenedesmus quadricauda (Turpin) Brebbison, Euglena granulata (Klebs) Schmitz and Gomphonema subtile Ehrenberg. The initial community composition was 2:2:1:1 for the four taxa respectively, which changed to 1.7:1.6:1:1 in pH 6.0, 1.6:2:1.6:1 in pH 5.5 and 1.2:2:1.8:1 in pH 5.0. The two chlorophytes showed almost gradual decreasing in growth rates and cell densities with the decreases in pH values, while diatom E. granulata showed more tolerance in a wide range of pH (6.5 to 5.5). Conspicuous changes in total biomass first observed in pH 6.0 and it reached the drastic level in pH 5.0. Moreover, significant changes in cell size of S. qaudricuada and E. granulata were found in pH 5.0 only. This lowered pH range has no effects on cell shape of the studied phytoplankton species.

Temperature-induced response in algal cell surface properties and behaviour: an experimental approach

Microalgae are considered an accurate indicator of ecosystem perturbations induced by global climate change. The present work aims to investigate the alteration of temperature on surface properties and behaviour of three algal species using the complementary surface methods (electrochemical and atomic force microscopy). The results showed that the temperature-induced response of algae is species-specific due to the structural features of the cell envelope. Wall-less algae experience the largest nanomechanical and chemical change, while algae with silicified walls show the pronounced chemical change in the degree of hydrophobicity. Alterations of surface properties suggest a molecular modification of the algal barrier and cytoskeletal rearrangements due to a change in cell size, while algal morphology reveals no change. The physiological activity of cells showed a different organisation of released extracellular substances in the form of fine fibrillar structures, aggregated particles, and dense networks. Both types of algal responses, physiological activity, and molecular modification of the cell barrier determine the cell adhesion and motility. This study highlights the role of surface properties in cell-substrate and cell–cell interactions, which is important for the understanding of algal behaviour at natural interfaces and the mechanism of algal biofilm and aggregate formation in aquatic systems under the stress.Graphical abstract

Simultaneous Measurement of Changes in Neutrophil Granulocyte Membrane Potential, Intracellular pH, and Cell Size by Multiparametric Flow Cytometry

Neutrophils provide rapid and efficient defense mechanisms against invading pathogens. Upon stimulation with proinflammatory mediators, including complement factors and bacterial peptides, neutrophils respond with changes in their membrane potential, intracellular pH, and cellular size. This study provides an approach to quantify these important changes simultaneously using multiparametric flow cytometry, thereby revealing a typical sequence of neutrophil activation consisting of depolarization, alkalization, and increase in cellular size. Additionally, the time resolution of the flow cytometric measurement is improved in order to allow changes that occur within seconds to be monitored, and thus to enhance the kinetic analysis of the neutrophil response. The method is appropriate for the reliable semiquantitative detection of small variations with respect to an increase, no change, and decrease in those parameters as demonstrated by the screening of various proinflammatory mediators. As a translational outlook, the findings are put into context in inflammatory conditions in vitro as well as in a clinically relevant whole blood model of endotoxemia. Taken together, the multiparametric analysis of neutrophil responsiveness regarding depolarization, alkalization, and changes in cellular size may contribute to a better understanding of neutrophils in health and disease, thus potentially yielding innovative mechanistic insights and possible novel diagnostic and/or prognostic approaches.

Red cell distribution width as a differential parameter between iron deficiency anemia and a-thalassemia: an empirical approach

Iron deficiency anemia (IDA) and thalassemia minor are the most common hypochromic microcytic anemias in the world. Different formulas have been proposed to differentiate IDA from beta thalassemia minor. However, yet no formula has been proposed to differentiate IDA from alpha thalassemia minor, and Hb electrophoresis is not helpful in this hemoglobinopathy. Red cell distribution width (RDW) as indicator of changes in red blood cell size is primarily employed to differentiate IDA from other microcytic anemias. An empirical approach involving iron therapy over 1 month has shown that an increase in Hb concentration by 1 g/dL over this period is indicative of IDA, while no changes in Hb concentration are suggestive of alpha thalassemia. RDW measured after iron therapy in order to differentiate IDA and related disorders from alpha thalassemia is a better index than an increased reticulocyte count. Due to the high prevalence of IDA and costly and time-consuming nature of specific diagnostic tests, the RDW index is considered as a very sensitive and cost-effective tool in the differential diagnosis of IDA.

Towards a Better Characterisation of Leukemic Cells in Chronic Lymphocytic Leukaemia: Cell-Size Heterogeneity Reflects Their Activation Status and Migratory Abilities.

Simple SummaryChronic lymphocytic leukaemia (CLL) is a heterogeneous chronic disease characterised by the clonal expansion of mature CD19+CD23+CD5+ B-cells in blood, bone marrow and lymphoid tissue. Despite the CLL tumour cell population showing considerable heterogeneity in cell size, the functional characteristics of leukemic cells that differ in size have not been explored. The results of our study demonstrate that differences in CLL cell size reflect their activation state, polarisation and migratory capacity, with large CLL cells being more activated, polarised and motile than the small CLL cells present in the CLL cell pool. Our data provide evidence of the importance of cell-size heterogeneity within the CLL cell pool and the dynamics of cell-size changes for disease pathogenesis.Chronic lymphocytic leukaemia (CLL) is a genetically, morphologically and phenotypically heterogeneous chronic disease with clinical variability between patients. Whether the significant heterogeneity of cell size within the CLL population contributes to the heterogeneous features of this disease has not been investigated. The present study aimed to characterise the phenotypic and functional properties of two subpopulations of typical CLL cells that differ in cell size: small (s-CLL) and large (l-CLL) CLL cells delineated by forward scatter cytometry. The s-CLL cells were characterised by the CD5lowCXCR4hi phenotype, while the l-CLL cells were characterised by the CD5hiCXCR4dim phenotype and indicated a higher expression of CXCR3, CD20, CD38 and HLA-DR. The l-CLL cells displayed higher migration activity towards CXCL12, a tendency towards a higher proliferation rate and an increased capacity to produce IgM in the presence of CpG compared with s-CLL cells. When stimulated with CpG and CXCL12, l-CLL cells were characterised by a higher polarisation phenotype and motility than s-CLL cells. Our study revealed that the differences in CLL cell size reflected their activation status, polarisation and migratory abilities. Our data provide evidence of the importance of cell-size heterogeneity within a CLL pool and the dynamics of cell-size changes for disease pathogenesis, thus deserving further investigation.

Thermal and Oxygen Flight Sensitivity in Ageing Drosophila melanogaster Flies: Links to Rapamycin-Induced Cell Size Changes.

Simple SummaryCold-blooded organisms can become physiologically challenged when performing highly oxygen-demanding activities (e.g., flight) across different thermal and oxygen environmental conditions. We explored whether this challenge decreases if an organism is built of smaller cells. This is because small cells create a large cell surface, which is costly, but can ease the delivery of oxygen to cells’ power plants, called mitochondria. We developed fruit flies in either standard food or food with rapamycin (a human drug altering the cell cycle and ageing), which produced flies with either large cells (no supplementation) or small cells (rapamycin supplementation). We measured the maximum speed at which flies were flapping their wings in warm and hot conditions, combined with either normal or reduced air oxygen concentrations. Flight intensity increased with temperature, and it was reduced by poor oxygen conditions, indicating limitations of flying insects by oxygen supply. Nevertheless, flies with small cells showed lower limitations, only slowing down their wing flapping in low oxygen in the hot environment. Our study suggests that small cells in a body can help cold-blooded organisms maintain demanding activities (e.g., flight), even in poor oxygen conditions, but this advantage can depend on body temperature.Ectotherms can become physiologically challenged when performing oxygen-demanding activities (e.g., flight) across differing environmental conditions, specifically temperature and oxygen levels. Achieving a balance between oxygen supply and demand can also depend on the cellular composition of organs, which either evolves or changes plastically in nature; however, this hypothesis has rarely been examined, especially in tracheated flying insects. The relatively large cell membrane area of small cells should increase the rates of oxygen and nutrient fluxes in cells; however, it does also increase the costs of cell membrane maintenance. To address the effects of cell size on flying insects, we measured the wing-beat frequency in two cell-size phenotypes of Drosophila melanogaster when flies were exposed to two temperatures (warm/hot) combined with two oxygen conditions (normoxia/hypoxia). The cell-size phenotypes were induced by rearing 15 isolines on either standard food (large cells) or rapamycin-enriched food (small cells). Rapamycin supplementation (downregulation of TOR activity) produced smaller flies with smaller wing epidermal cells. Flies generally flapped their wings at a slower rate in cooler (warm treatment) and less-oxygenated (hypoxia) conditions, but the small-cell-phenotype flies were less prone to oxygen limitation than the large-cell-phenotype flies and did not respond to the different oxygen conditions under the warm treatment. We suggest that ectotherms with small-cell life strategies can maintain physiologically demanding activities (e.g., flight) when challenged by oxygen-poor conditions, but this advantage may depend on the correspondence among body temperatures, acclimation temperatures and physiological thermal limits.

IVEN: A quantitative tool to describe 3D cell position and neighbourhood reveals architectural changes in FGF4-treated preimplantation embryos.

Architectural changes at the cellular and organism level are integral and necessary to successful development and growth. During mammalian preimplantation development, cells reduce in size and the architecture of the embryo changes significantly. Such changes must be coordinated correctly to ensure continued development of the embryo and, ultimately, a successful pregnancy. However, the nature of such transformations is poorly defined during mammalian preimplantation development. In order to quantitatively describe changes in cell environment and organism architecture, we designed Internal Versus External Neighbourhood (IVEN). IVEN is a user-interactive, open-source pipeline that classifies cells into different populations based on their position and quantifies the number of neighbours of every cell within a dataset in a 3D environment. Through IVEN-driven analyses, we show how transformations in cell environment, defined here as changes in cell neighbourhood, are related to changes in embryo geometry and major developmental events during preimplantation mammalian development. Moreover, we demonstrate that modulation of the FGF pathway alters spatial relations of inner cells and neighbourhood distributions, leading to overall changes in embryo architecture. In conjunction with IVEN-driven analyses, we uncover differences in the dynamic of cell size changes over the preimplantation period and determine that cells within the mammalian embryo initiate growth phase only at the time of implantation.

Cellular Property of the Detonation Wave of Pre-mixed Methane-Oxygen in Annular Tube

In order to investigate the detonation propagation in the annular tube, the experiments are conducted with methane-oxygen mixture. Annual tubes with different inner diameter D0 were configured and smoked foils were fixed on both outer wall and the inner wall of the annular tube to record the detonation structure. Firstly, it is found that transverse wave modes on both outer wall and the inner wall of the annular tube increase with increasing initial pressure and inner diameter of the channel. In addition, the measurement of angle between the transverse wave and the tube axis disagrees acoustic theory within the error range. In the results, the angle and the cell size changes less with pressure and more with different position of the channel, demonstrating the strongly dependence of the tube geometric. What’s more, the trajectories of right-running transverse wave were traced and some of the transverse waves disappeared in the polycarbonate tube, because the initial energy of different transverse waves are different and the energy required to sustain the transverse vibration of spinning detonation must be derived from the chemical energy release at the front.

High content genome-wide siRNA screen to investigate the coordination of cell size and RNA production

Coordination of RNA abundance and production rate with cell size has been observed in diverse organisms and cell populations. However, how cells achieve such ‘scaling’ of transcription with size is unknown. Here we describe a genome-wide siRNA screen to identify regulators of global RNA production rates in HeLa cells. We quantify the single-cell RNA production rate using metabolic pulse-labelling of RNA and subsequent high-content imaging. Our quantitative, single-cell measurements of DNA, nascent RNA, proliferating cell nuclear antigen (PCNA), and total protein, as well as cell morphology and population-context, capture a detailed cellular phenotype. This allows us to account for changes in cell size and cell-cycle distribution (G1/S/G2) in perturbation conditions, which indirectly affect global RNA production. We also take advantage of the subcellular information to distinguish between nascent RNA localised in the nucleolus and nucleoplasm, to approximate ribosomal and non-ribosomal RNA contributions to perturbation phenotypes. Perturbations uncovered through this screen provide a resource for exploring the mechanisms of regulation of global RNA metabolism and its coordination with cellular states.

Concerted evolution of body mass, cell size and metabolic rate among carabid beetles

Alterations in cell number and size are apparently associated with the body mass differences between species and sexes, but we rarely know which of the two mechanisms underlies the observed variance in body mass. We used phylogenetically informed comparisons of males and females of 19 Carabidae beetle species to compare body mass, resting metabolic rate, and cell size in the ommatidia and Malpighian tubules. We found that the larger species or larger sex (males or females, depending on the species) consistently possessed larger cells in the two tissues, indicating organism-wide coordination of cell size changes in different tissues and the contribution of these changes to the origin of evolutionary and sex differences in body mass. The species or sex with larger cells also exhibited lower mass-specific metabolic rates, and the interspecific mass scaling of metabolism was negatively allometric, indicating that large beetles with larger cells spent relatively less energy on maintenance than small beetles. These outcomes also support existing hypotheses about the fitness consequences of cell size changes, postulating that the low surface-to-volume ratio of large cells helps decrease the energetic demand of maintaining ionic gradients across cell membranes. Analyses with and without phylogenetic information yielded similar results, indicating that the observed patterns were not biased by shared ancestry. Overall, we suggest that natural selection does not operate on each trait independently and that the linkages between concerted cell size changes in different tissues, body mass and metabolic rate should thus be viewed as outcomes of correlational selection.

Suppression of mTOR Signaling Attenuates Apoptotic Effects Induced By the NEDD8-Activating Enzyme Inhibitor Pevonedistat (MLN4924) in Acute Myeloid Leukemia Cells

Acute myelogenous leukemia (AML) is a heterogeneous disease and often relapses after standard chemotherapy. Therefore, novel therapies for AML are needed. Recently, the NEDD8 conjugation pathway and the mammalian target of rapamycin (mTOR) signaling pathway have emerged as important regulatory pathways for cancer therapy. Since some components of the mTORC1/2 pathways such as Deptor are neddylation targets, we postulated that combined exposure to NEDD8-activating enzyme (NAE) inhibitor pevonedistat and the dual mTORC1/2 inhibitor sapanisertib (TAK-228) might have additive or synergistic effects in AML.To test pevonedistat and sapanisertib effects on AML cells, we used the agents alone or in combination followed by assessment of proliferation, apoptosis and signal transduction in human AML cell lines as well as in primary cells. By AnnexinV/7AAD flow cytometry analysis, we found that pevonedistat significantly induced AML cell death and apoptosis in a time- and dose-dependent manner. Western blot showed that poly (ADP-ribose) polymerase (PARP) and caspases 3/8/9 were cleaved, indicating that pevonedistat-treated cells were actively undergoing apoptosis. Among the six AML cell lines (HL-60, KG1a, Molm-13, MV4-11, THP-1 and U937) tested, MV4-11 was the most sensitive cell line (IC50 <25nM) and KG1a was the least sensitive cell line (IC50 >1000nM) after 48 hours of exposure. The average IC50 on primary AML blast cells (5 primary samples) was 400nM at 48 hours. Western blot analysis showed that the neddylated cullin levels were markedly decreased confirming inhibition of the NEDD8 conjugation pathway by pevonedistat. We also found that pevonedistat induced morphologic changes and increased CD11b expression in Molm-13 and MV4-11 cells, indicating cell differentiation.The effects of pevonedistat on AML cells could be attenuated by pre- or co-treatment with the mTORC1/2 inhibitor sapanisertib. We found that sapanisertib alone induced limited cell death in most AML cell lines and primary AML patient blasts. The results from XTT assay and flow cytometry analysis indicated that sapanisertib alone significantly decreased cell size, arrested cells in G0-G1 phase and reduced protein content of AML cells but did not induce robust apoptosis. This suggested that through inactivating mTOR signal pathway, sapanisertib suppressed cell activity and induced a dormant/quiescent status and thereby attenuated the apoptotic effects of pevonedistat. With concurrent pevonedistat and sapanisertib exposure, the expression of apoptotic mediators such as cleaved PARP and caspases 3 were decreased compare to pevonedistat treatment alone (See Figure).To confirm the role of mTOR inhibition on pevonedistat-induced AML apoptosis, we targeted mTORC1/2 by knocking down mTOR expression with small hairpin RNAs (shRNA). Downregulation of mTOR expression mirrored the effects of sapanisertib. mTOR inhibition by shRNAs resulted in decreased AML cell size and arrest of cells in G0-G1 phase. In addition, pevonedistat-induced cell death was significantly impeded in the cells infected with mTOR shRNA lentivirus compared to scrambled shRNA (p<0.01). Therefore, we validated that mTOR signal inactivation protects AML cells from NAE inhibitor pevonedistat-induced apoptosis using both pharmaceutical and genetic approaches. To further study the mechanism behind mTOR inhibition inducing resistance to pevonedistat, we tested rapamycin, an mTORC1 inhibitor. Concurrent exposure to rapamycin did not significantly alter the effects of pevonedistat and did not change cell size and cell cycle status. In addition, rapamycin did not inhibit the phosphorylation of 4E-BP1, suggesting that mTORC1 inhibition alone may be insufficient to inhibit the effects of pevonedistat, and suggesting a role for mTORC2.Taken together, our data demonstrate that the NEDDylation pathway inhibitor pevonedistat has strong potential as a therapeutic agent for AML therapy via apoptosis induction. However, when AML cells are arrested in G0-G1 with reduced proliferation rates by dual mTORC1/2-inhibition, the ability of pevonedistat to induce apoptosis is suppressed. Whether this is due solely to the cytostatic effects of dual mTOR inhibitors or also involves other mechanisms remains uncertain. While both sapanisertib and pevonedistat have clear anti-leukemia effects, their concurrent use may inhibit NAE inhibitor-induced apoptosis. [Display omitted] DisclosuresLiesveld:Onconova: Honoraria; Seattle Genetics: Honoraria.

Mesenchymal Stromal Cell Sialylation Enhances Immune Suppression in Multiple Myeloma

Introduction: In multiple myeloma (MM) the bone marrow (BM) microenvironment, plays an important role in mediating survival, proliferation, resistance to therapy as well as disease evolution. The BM microenvironment is mainly composed of mesenchymal stromal cells (BM-MSC), endothelial cells, immune cells, and extracellular matrix. BM-MSC can acquire distinct functional phenotypes that are influenced by signals in the microenvironment and have been shown to facilitate MM development and drug resistance through cell-cell contact and cytokine secretion. However, little is known about the mechanisms of immune modulation mediated by BM-MSC in the inflammatory MM tumour microenvironment (TME). Aberrant glycosylation is a hallmark of cancer cells, playing an important role in tumor progression. Our data strongly links aberrant sialylation with poor prognosis in MM (1) but to date the immunological consequences of this phenomenon have not been explored. Here we sought to investigate if regulation of BM-MSC sialylation alters their ability to inhibit effector T-cell function in an inflammatory microenvironment, characteristic of the MM TME.Methods: BM-MSC were isolated from BALB/c mice and extensively characterized in vitro . Since inflammatory mediators, such as TNF-α and IL-1β are present at high levels in MM due to constitutive NF-kappaB activity, we assessed the effects of both of these pro-inflammatory mediators on BM-MSC phenotype. BM-MSC were treated with both tumor necrosis factor alpha (TNF-α 50ng/ml) and interleukin 1 beta (IL-1β 50ng/ml) (iMSC) for 72 hours and cell surface sialylation was analysed by flow cytometry. To determine immunoregulatory ability, BM-MSC and iMSC were co-cultured in mixed lymphocyte reactions (MLRs) for 96 hours. T-cell proliferation, activation, death and differentiation were determined by flow cytometry. To determine the role of increased sialylation in lymphocyte suppression, both BM-MSC and iMSC were pre-treated with a sialyltransferase inhibitor (3Fax-Neu5Ac) for 72 hours prior to TNF-α and IL-1β stimulation. To determine the important of antigen presenting cells in sialic acid dependent regulation of T cell phenotype, CD11c+ and CD11b+ innate immune cells were removed via MACS sorting prior to co-culture. BM-MSC: Lymphocyte co-culture supernatants for nitric oxide (NO), IL-10 and PGE-2 were analysed by ELISA.Results: iMSC displayed significant phenotypical changes with CD73 and CD44 increasing significantly. Both SCA-1 and CD105 were significantly reduced. No changes in cell viability were noted after pre-treatment and no significant changes in proliferation were observed. iMSC had significantly increased levels of both α2-3 and α2-6 linked sialic acid when compared to BM-MSC. In vitro, iMSC displayed an enhanced ability to inhibit lymphocyte proliferation when compared to BM-MSC alone. iMSC significantly inhibited the proliferation of hyper-stimulated CD3+CD4+ and CD3+CD8+ lymphocytes. 3Fax-Neu5Ac treated BM-MSC and iMSC displayed no significant changes in phenotype, viability, proliferation and cell size. However, following sialyltransferase inhibition, iMSC lost the ability to suppress both CD3+CD4+ and CD3+CD8+ lymphocytes resulting in significant restoration of lymphocyte proliferation. This restoration of proliferation was further enhanced in the absence of CD11c+ and CD11b+ innate immune cells. Cell surface sialic acid expression correlated with both CD3+CD4+ and CD3+CD8+ lymphocyte suppression. No changes in soluble mediators of BM-MSC immunomodulation, such as nitic oxide, IL-10 and PGE-2 were observed suggesting that restoration in proliferation was both sialic acid and cell-contact dependent.Conclusions: Our findings confirm inflammation, characteristic of the MM TME, induces BM-MSC sialylation and enhances their ability to suppress activated adaptive and innate immune effectors. Understanding the potential utility of targeting BM-MSC sialylation and consequently their immunomodulatory potential, may enhance immune cell activation in the MM microenvironment, providing further rationale to targeting BM-MSC sialylation in the context of MM. We suggest that understanding the functional importance of the BM stroma and its interaction with MM and immune cells is likely to lead to the identification of sialylation, as a new molecular target. 1.Glavey SV, et al Blood. 2014 Sep 11;124(11):1765-76 [Display omitted] DisclosuresRyan:Janssen: Research Funding. O'Dwyer:Onkimmune Ltd: Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties; GlycoMimetics Inc: Research Funding; Janssen: Consultancy, Honoraria, Research Funding.

Quantitative trait loci (QTL) analysis of leaf related traits in spinach (Spinacia oleracea L.)

BackgroundSpinach (Spinacia oleracea L.) is an important leafy vegetable crop, and leaf-related traits including leaf length, leaf width, and petiole length, are important commercial traits. However, the underlying genes remain unclear. The objective of the study was to conduct QTL mapping of leaf-related traits in spinach.ResultsA BC1 population was used to construct the linkage map and for QTL mapping of leaf length, leaf width, petiole length, and the ratio of leaf length to width in 2015 and 2019. Two genetic linkage maps were constructed by specific locus amplified fragment sequencing (SLAF-seq), and kompetitive allele specific PCR (KASP) technology, respectively using BC1 population in 2015. Based on the results of 2015, the specific linkage groups (LG) detected QTLs were generated using BC1 population in 2019. A total of 13 QTLs were detected for leaf-related traits, only five QTLs being repeatedly detected in multiple years or linkage maps. Interestingly, the major QTLs of leaf length, petiole length, and the ratio of leaf length to width were highly associated with the same SNP markers (KM3102838, KM1360385 and KM2191098). A major QTL of leaf width was mapped on chromosome 1 from 41.470−42.045 Mb. And 44 genes were identified within the region. Based on the GO analysis, these genes were significantly enriched on ribonuclease, lyase activity, phosphodiester bond hydrolysis process, and cell wall component, thus it might change cell size to determine leaves shape.ConclusionsFive QTLs for leaf-related traits were repeatedly detected at least two years or linkage maps. The major QTLs of leaf length, petiole length, and the ratio of leaf length to width were mapped on the same loci. And three genes (Spo10792, Spo21018, and Spo21019) were identified as important candidate genes for leaf width.

Exploring the antifungal activity and mechanism of action of Zingiberaceae rhizome extracts against Malassezia furfur

Ethnopharmacological relevanceRhizomes from members of Zingiberaceae have long been used in Thai traditional medicine to treat cutaneous fungal infections, including Malassezia-related skin disorders. Alpinia galanga, Curcuma longa, Zingiber cassumunar, and Zingiber officinale are particularly popular in folk remedies. Aim of the studyOn account of the application background in traditional medicine, the present study aims to screen and determine the composition and possible mechanism of the rhizome extracts of selected Zingiberaceae and corresponding fractions against M. furfur. Materials and methodsAll solvent extracts (ethanol, methanol, and n-hexane) obtained from each plant were screened for anti-Malassezia activity by agar disc diffusion assay. The MIC and MFC values of the potent rhizome extract and its bioactive fraction isolated by TLC were determined using broth dilution assay followed by chemical characterization using GC-MS. The anti-Malassezia mechanism was investigated by macroscopic and microscopic observation of cells grown in the yeast phase and hyphal phase. ResultsThe primary screening results showed that the n-hexane extract from A. galanga possessed the most significant anti-Malassezia activity. The MIC and MFC values of this extract were in a range of 0.04–0.08 mg/mL and 0.04–0.16 mg/mL, respectively. The TLC purification of the n-hexane extract from A. galanga gave a total of nine fractions, of which only a single exhibited anti-Malassezia activity. The GC-MS analysis of the rhizome extract and the derivative fraction revealed that the major constituents were (2,6-dimethylphenyl)borate followed by a trace content of 1,8-cineol and hydrocarbons. For the antifungal mechanism of the fraction, treatments of the fraction led to morphological changes in cell size and shape, exerted massive vacuoles in yeast form, and inhibited the transition to hyphae but not likely affected chitin contents of the cell wall of M. furfur. ConclusionsAccording to the results, the n-hexane extract of A. galanga rhizome exhibits promising anti-Malassezia potential. The inhibitory effect on virulent hyphal growth supports that A. galanga is a valuable source of natural antifungal agents for further pharmaceutical research.

Evaluation of SnS:Cu Thin Film Properties Obtained by USP Technique to Implement It as an Absorbent Layer in Solar Cells Using SCAPS

Tin sulfide doped with copper (SnS:Cu) thin films were deposited on glass substrates by the ultrasonic spray pyrolysis (USP) technique at different concentration ratios (y = [Cu]/[Sn] = 0% (undoped), 2%, 5% and 10%). The aim of this work is to analyze the effect of copper on structural, morphological, and optoelectronic properties of SnS:Cu and discuss their possible application as an absorber layer in a solar cell structure proposed which is simulated using SCAPS software. X-ray diffraction (XRD) reveals an orthorhombic structure in the undoped sample and a cubic structure in doped ones. Raman spectroscopy suggests a possible unit cell size change due to the addition of Cu. Scanning electron microscopy (SEM) shows growth in grain density with an increasing y. Image analysis based on second-order features was used to discuss grain distribution. UV-VIS spectroscopy helps to find an increase of bandgap for the doped samples when copper concentration increases, going from 1.82 eV in the doped film y = 2% to 2.2 eV in the 10% doped samples. A value of 3.51 eV was found for the undoped sample y = 0%. A rise in both carrier concentration and mobility but a decrease in resistivity when y is increased was observed through the Hall–Van der Pauw technique. Simulations by SCAPS helped conclude that considering the material thickness, the SnS:Cu compound can be an alternative for implementation in the manufacturing of solar cells as an absorber layer since it is possible to obtain the optoelectronic properties necessary using the UPS economical technique.

Macroscopic Lane Change Model—A Flexible Event-Tree-Based Approach for the Prediction of Lane Change on Freeway Traffic

Binary logistic regression has been used to estimate the probability of lane change (LC) in the Cell Transmission Model (CTM). These models remain rigid, as the flexibility to predict LC for different cell size configurations has not been accounted for. This paper introduces a relaxation method to refine the conventional binary logistic LC model using an event-tree approach. The LC probability for increasing cell size and cell length was estimated by expanding the LC probability of a pre-defined model generated from different configurations of speed and density differences. The reliability of the proposed models has been validated with NGSIM trajectory data. The results showed that the models could accurately estimate the probability of LC with a slight difference between the actual LC and predicted LC (95% Confidence Interval). Furthermore, a comparison of prediction performance between the proposed model and the actual observations has verified the model’s prediction ability with an accuracy of 0.69 and Area Under Curve (AUC) value above 0.6. The proposed method was able to accommodate the presence of multiple LCs when cell size changes. This is worthwhile to explore the importance of such consequences in affecting the performance of LC prediction in the CTM model.

Real-Time Deformability Cytometry Detects Leukocyte Stiffening After Gadolinium-Based Contrast Agent Exposure.

Reports on gadolinium (Gd) retention in soft tissues after administration of Gd-based contrast agents (GBCAs) raise concerns about Gd-induced changes in the biophysical properties of cells and tissues. Here, we investigate if clinical GBCAs of both classes of linear and macrocyclic structure cause changes in the mechanical properties of leukocytes in human blood samples. Real-time deformability cytometry was applied to human blood samples from 6 donors. The samples were treated with 1 mM gadoteric acid (Dotarem), gadopentetic acid (Magnevist), gadobutrol (Gadovist), or Gd trichloride at 37°C for 1 hour to mimic clinical doses of GBCAs and exposure times. Leukocyte subtypes-lymphocytes, monocytes, and neutrophils-were identified based on their size and brightness and analyzed for deformability, which is inversely correlated with cellular stiffness. We observed significant stiffening (3%-13%, P < 0.01) of all investigated leukocyte subtypes, which was most pronounced for lymphocytes, followed by neutrophils and monocytes, and the effects were independent of the charge and steric structure of the GBCA applied. In contrast, no changes in cell size and brightness were observed, suggesting that deformability and cell stiffness measured by real-time deformability cytometry are sensitive to changes in the physical phenotypes of leukocytes after GBCA exposure. Real-time deformability cytometry might provide a quantitative blood marker for critical changes in the physical properties of blood cells in patients undergoing GBCA-enhanced magnetic resonance imaging.

Blockade of the CD6-ALCAM pathway modulates effector T cell function

Abstract CD6 is a T-cell costimulatory receptor predominantly expressed on T cells and promotes immune synapse formation, T-cell activation, and T-cell migration via interaction with activated leukocyte cell adhesion molecule (ALCAM). The CD6-ALCAM pathway has been linked to the pathogenesis of multiple autoimmune and inflammatory diseases. While the contribution of CD6 to T cell activation has been well described, less is known regarding the expression levels and role of CD6 on effector and memory T cells which contribute to ongoing disease. Consequently, the aim of this study was to determine the role of CD6 specifically on Th1, Th2 and Th17 effector T cells (Teff). Blockade of the CD6 pathway using itolizumab, a humanized mAb currently being tested in the clinic, during restimulation of fully differentiated Th1, Th2 or Th17 Teff cells in the presence of ALCAM inhibited multiple effector functions, including proliferation and changes in cell size. An average 40% decrease in proliferation was observed across multiple donors. Furthermore, treatment of Teff cells with itolizumab resulted in a significant decrease in relevant cytokines such as IFNγ and TNFα for Th1, IL-4 and IL-13 for Th2, TNFα and IL-17 for Th17, as well as in expression levels of T cell markers of activation and exhaustion such as CD69, CD25, and PD-1. This effect was exclusively observed in the presence of ALCAM, indicating that the effect was specific to blockade of the CD6-ALCAM pathway. This study is the first to characterize the CD6-ALCAM pathway as a key regulator of effector Teff cell function. As these are the cells that drive disease pathogenicity, these data support targeting the CD6-ALCAM pathway to inhibit effector Teff cell populations in autoimmune and inflammatory diseases.