Plating Efficiency Research Articles

압력용기 클래드 보수용 전해니켈도금 인자 관계 연구

Pure nickel with a thickness of 1 mm was plated on type 304 stainless steels and low alloy steels (JIS G3131 SPHC) by electrolytic plating method in a circulating plating bath. Plating performance, mechanical properties, and surface characteristics were evaluated in terms of pretreatment process, anode material, pH, current density, and flow rate of the plating solution. Addition of hydrochloric acid during pre-treatment process improved the adhesion performance of plating. To improve plating efficiency, it is desirable to use S-nickel rather than electrolytic nickel. The use of S-nickel was also confirmed to be desirable for maintaining the pH and concentration of the plated solution. The defect of the plating using S-nickel anode produced pit on the surface. However, it is believed that proper control can be obtained by increasing the flow rate. Internal stress and hardness values of electrolytic nickel plating according to current density need to be carried out with further studies.

Antiviral and Virucidal Activities of Umesu Phenolics on Influenza Viruses.

In this study, umesu phenolics were purified from the salt extracts of Japanese apricot (Nanko-mume cultivar of Prunus mume Sieb. et Zucc.). Characterization of umesu phenolics revealed that, when added to the culture media of the infected cells, they inhibited the multiplication of influenza and many other RNA and DNA viruses. In addition to these antiviral activities, the phenolics significantly decreased the plating efficiency of influenza virus, if present in the virus inoculum. More drastic effects were observed in terms of virucidal activity; the infectivity of several strains of influenza viruses decreased less than 0.001 when they were incubated with 4 mg/ml phenolics at 30 ℃ for 5 min. The virucidal activity of phenolics was found to be more remarkable in acidic conditions; however, the activity was not merely a result of the acidity of the phenolics. These results clearly support the antiviral and virucidal activities of the umesu phenolics against influenza viruses and suggest their potential pharmacological usefulness as disinfectants or preventive medicine against superficial infections, such as the respiratory infections.

Phenotypic and Genetic Predictors of Pathogenicity and Virulence in Flavobacterium psychrophilum.

Flavobacterium psychrophilum causes bacterial cold-water disease (BCWD) in farmed rainbow trout (Oncorhynchus mykiss), with the multilocus sequence typing (MLST) clonal complex (CC) CC-ST10 accounting for the majority of outbreaks globally. The development of alternative strategies to antibiotic treatment of BCWD using bacteriophage-based control of F. psychrophilum, or virulence factors as targets for therapy, requires knowledge of the phage-sensitivity of outbreak strains and of universal traits contributing to their pathogenicity. To examine the association between virulence and both genetic (MLST sequence type (ST) and PCR-serotype) and phenotypic characteristics (adherence, antibiotic resistance, colony spreading motility, hemolytic and proteolytic activity), the median lethal dose (LD50) of 26 geographically disparate F. psychrophilum isolates was determined in rainbow trout. Furthermore, the in vitro sensitivity of the isolates against five bacteriophages was determined by the efficiency of plating (EOP). The tested F. psychrophilum isolates were mainly represented by CC-ST10 genotypes (22 out of 26) and showed up to 3-log differences in LD50 (8.9 × 103 to 3.1 × 106 CFU). No association between MLST ST and virulence was found because of a high variation in LD50 within STs. All identified serotypes (0, 1, and 2) were pathogenic, but ten most virulent isolates belonged to serotype 1 or 2. Isolates of high (LD50 < 105 CFU), moderate (LD50 = 105–106 CFU), and weak (LD50 > 106 CFU) virulence were similar in phenotypic characteristics in vitro. However, the only non-virulent CC-ST10 isolate was deficient in spreading motility and proteolytic activity, indicating that the characteristics are required for pathogenicity in F. psychrophilum. Univariate correlation studies found only non-significant associations between LD50 and the measured phenotypic characteristics, and the multivariable analysis did neither reveal any significant predictors of virulence. The majority of isolates (16 out of 26) were sensitive to at least four bacteriophages, with up to a 6-log variation in the EOP. Most CC-ST10 isolates (16 out of 22) were sensitive to the examined phages, including 5 out of the 7 most virulent isolates represented by prevalent and antibiotic-resistant STs. Our findings suggest that control of BCWD using lytic phages or interventions targeting shared characteristics of pathogenic F. psychrophilum strains should be further explored.

Effects of combined 5-Fluorouracil and ZnO NPs on human breast cancer MCF-7 Cells: P53 gene expression, Bcl-2 signaling pathway, and invasion activity

Objective(s): The significant contribution of nanoparticles to cancer treatment has attracted therapeutic attention. The present study aimed to evaluate the synergistic effects of 5-fluorouracil (5-FU) and zinc oxide nanoparticles (ZnO NPs) as multimodal drug delivery on human breast cancer MCF-7 cells.Materials and Methods: In this in-vitro study, the impact of 5-FU and ZnO NPs in the single or combined forms was evaluated on cell viability, colony formation, apoptosis, p53 gene expression, and Bcl-2 signaling protein in MCF-7 breast cancer cell line using several techniques, such as MTT, clonogenic assay, flow cytometry, real-time quantitative polymerase chain reaction, and Western blot.Results: In this study, 5-FU combined with ZnO NPs showed synergistic effects against MCF-7 within 48 hours. In addition, the combination of 5-FU and ZnO NPs at the respective concentrations of 1 µM and 45 µg/ml exhibited significant apoptosis (79.53), p53 gene expression (3.6 folds), reduction of cell invasion (9.82), and plating efficiency (5), thereby leading to the significant reduction of cell viability (40±0.9) and decreased Bcl-2 anti-apoptotic protein relative to untreated control cells. Conclusion: According to the results, the synergistic effects of combined ZnO NPs and 5-FU on MCF-7 human breast cancer cells were exerted via Bcl-2 inhibition and the up-regulation of p53 expression.

Two Phages of the Genera Felixounavirus Subjected to 12 Hour Challenge on Salmonella Infantis Showed Distinct Genotypic and Phenotypic Changes.

Salmonella Infantis is considered in recent years an emerging Salmonella serovar, as it has been associated with several outbreaks and multidrug resistance phenotypes. Phages appear as a possible alternative strategy to control Salmonella Infantis (SI). The aims of this work were to characterize two phages of the Felixounavirus genus, isolated using the same strain of SI, and to expose them to interact in challenge assays to identify genetic and phenotypic changes generated from these interactions. These two phages have a shared nucleotide identity of 97% and are differentiated by their host range: one phage has a wide host range (lysing 14 serovars), and the other has a narrow host range (lysing 6 serovars). During the 12 h challenge we compared: (1) optical density of SI, (2) proportion of SI survivors from phage-infected cultures, and (3) phage titer. Isolates obtained through the assays were evaluated by efficiency of plating (EOP) and by host-range characterization. Genomic modifications were characterized by evaluation of single nucleotide polymorphisms (SNPs). The optical density (600 nm) of phage-infected SI decreased, as compared to the uninfected control, by an average of 0.7 for SI infected with the wide-host-range (WHR) phage and by 0.3 for SI infected with the narrow-host-range (NHR) phage. WHR phage reached higher phage titer (7 × 1011 PFU/mL), and a lower proportion of SI survivor was obtained from the challenge assay. In SI that interacted with phages, we identified SNPs in two genes (rfaK and rfaB), which are both involved in lipopolysaccharide (LPS) polymerization. Therefore, mutations that could impact potential phage receptors on the host surface were selected by lytic phage exposure. This work demonstrates that the interaction of Salmonella phages (WHR and NHR) with SI for 12 h in vitro leads to emergence of new phenotypic and genotypic traits in both phage and host. This information is crucial for the rational design of phage-based control strategies.

Characterization of a bacteriophage with broad host range against strains of Pseudomonas aeruginosa isolated from domestic animals

BackgroundPseudomonas aeruginosa is an opportunistic pathogen and one of the leading causes of nosocomial infections. Moreover, the species can cause severe infections in cystic fibrosis patients, in burnt victims and cause disease in domestic animals. The control of these infections is often difficult due to its vast repertoire of mechanisms for antibiotic resistance. Phage therapy investigation with P. aeruginosa bacteriophages has aimed mainly the control of human diseases. In the present work, we have isolated and characterized a new bacteriophage, named Pseudomonas phage BrSP1, and investigated its host range against 36 P. aeruginosa strains isolated from diseased animals and against P. aeruginosa ATCC strain 27853.ResultsWe have isolated a Pseudomonas aeruginosa phage from sewage. We named this virus Pseudomonas phage BrSP1. Our electron microscopy analysis showed that phage BrSP1 had a long tail structure found in members of the order Caudovirales. “In vitro” biological assays demonstrated that phage BrSP1 was capable of maintaining the P. aeruginosa population at low levels for up to 12 h post-infection. However, bacterial growth resumed afterward and reached levels similar to non-treated samples at 24 h post-infection. Host range analysis showed that 51.4% of the bacterial strains investigated were susceptible to phage BrSP1 and efficiency of plating (EOP) investigation indicated that EOP values in the strains tested varied from 0.02 to 1.72. Analysis of the phage genome revealed that it was a double-stranded DNA virus with 66,189 bp, highly similar to the genomes of members of the genus Pbunavirus, a group of viruses also known as PB1-like viruses.ConclusionThe results of our “in vitro” bioassays and of our host range analysis suggested that Pseudomonas phage BrSP1 could be included in a phage cocktail to treat veterinary infections. Our EOP investigation confirmed that EOP values differ considerably among different bacterial strains. Comparisons of complete genome sequences indicated that phage BrSP1 is a novel species of the genus Pbunavirus. The complete genome of phage BrSP1 provides additional data that may help the broader understanding of pbunaviruses genome evolution.

PS1231 ACTIVATION OF AUTOPHAGY IN ENDOTHELIAL CELLS ENHANCES HEMATOPOIETIC STEM CELLS

Background:Emerging murine studies showed that endothelial cells (ECs) serve as an instructive platform to nurture the hematopoietic stem cell (HSCs). Autophagy, the self‐degradative process responsible for the elimination of cytosolic components, has recently been demonstrated to have important effects upon EC function. But no previous study had explore the role of autophagy in the process of ECs to support HSCs. Our previous work reported that abnormal bone marrow (BM) ECs were involved in the pathogenesis of poor graft function (PGF), a serious complication with pancytopenia after allogeneic hematopoietic stem cell transplantation (allo‐HSCT), and improving dysfunctional BM ECs could promote hematopoiesis ( BBMT 2013; BMT 2016; Blood 2016), but the explicit mechanism need to be further clarified. Given the important role of autophagy in ECs, we hypothesized that up‐regulation of autophagy in ECs might enhance HSCs.Aims:The current study was performed to explore whether autophagy would affect the ability of ECs to support HSCs. Moreover, to evaluate whether autophagy in BM ECs play a role in the pathogenesis of PGF, and investigate the effects of autophagic regulation on ECs and thereby promoting hematopoiesis.Methods:In vitro regulation of autophagy by rapamycin (the autophagy activators) or hydroxychloroquine (HCQ, the autophagy inhibitor) were administrated to HUVECs. To explore whether autophagy would affect the ability of ECs to support HSCs in vitro, BM CD34+ cells from healthy donors were cocultured with cultivated BM ECs. Colony‐forming unit (CFU) and the apoptosis of co‐cultured HSCs were analyzed. Subsequently, a prospective case‐control study was conducted to evaluate the autophagy levels in BM ECs and CFU plating efficiency in HSCs from PGF patients, and their matched good graft function (GGF) patients. The expression levels of autophagy‐related markers (LC3, Beclin1, and P62), and intracellular autophagosomes were detected by immunohistochemical staining, flow cytometry, western blot and transmission electron microscopy. Furthermore, to explore whether autophagic regulations would affect the number and ability of BM ECs from PGF patients, including the ability to support HSCs in vitro.Results:In vitro upregulation of autophagy by rapamycin quantitatively and functionally improved HUVECs, which could further significantly improve CFU plating efficiency in BM HSCs co‐cultured with HUVECs, whereas inhibition of autophagy by HCQ produce opposite effects. In the prospective case‐control study, defective autophagy was found in BM ECs from PGF patients, characterized by decreased intracellular autophagosomes and autophagic vacuoles, decreased expression of LC3‐II and Beclin1, and high level of P62, when compared to GGF patients. The co‐culture of BM CD34+ cells with BM ECs showed significant deficient CFU plating efficiency, and increased apoptosis of CD34+ cells in PGF patients. Subsequently, in vitro upregulation of autophagy by rapamycin quantitatively and functionally improved BM ECs from PGF patients. Furthermore, rapamycin significant improved CFU plating efficiency, and decreased apoptosis in BM HSCs co‐cultured with BM ECs from PGF patients.Summary/Conclusion:Taken together, our results suggest that activation of autophagy in ECs could enhance their HSCs‐supporting capacity which indicating a new mechanism of HSC regulation. Moreover, defective autophagy may be involved in the pathogenesis of PGF, and rapamycin might provide a novel clinical intervention for PGF patients post‐HSCT in the future.

Dynamics of Lithium Deposition in Li-S Batteries: An Elucidation with a Lithium Sulfide Cathode and a Bare Copper-Foil Current Collector without Any Free Lithium Metal

The realization of high-energy density lithium-sulfur batteries requires the development of stable, reversible, and practical lithium-metal anodes. This necessitates a comprehensive study of the dynamics of lithium deposition in Li-S batteries. Most of the research in Li-S batteries has focused on half-cell studies, which include a thick metallic-lithium chip as the anode. With an infinite supply of lithium, this precludes understanding the impact of lithium degradation and loss on electrochemical performance. Hence, an investigation of Li-S cell performance in the absence of a large excess of lithium, i.e., under lithium-limited conditions, is acutely needed. To this end, a novel full-cell configuration for Li-S batteries is presented here that comprises a lithium sulfide cathode paired with a bare copper or nickel foil as the hostless-anode substrate for lithium deposition. As the cell has only a limited and stoichiometric amount of lithium from the cathode without any free lithium metal on the anode, its electrochemical performance is limited by the efficiency of lithium plating and stripping. This allows for a more realistic assessment of Li-S cell performance. In addition, the role and dynamical behavior of the lithium anode can be effectively clarified by contrasting the electrochemical performance of the full cell to a comparable half cell. Counter-intuitively, the Li2S full cell shows only a modest decline in electrochemical performance in comparison to the Li2S half cell. It demonstrates reasonable capacity retention with an average Coulombic efficiency of 96% over 100 cycles. In contrast, the capacity of a similar lithium-plating-based full cell employing LiFePO4 cathodes declines to near zero within 10 cycles. The Coulombic efficiency of plating and stripping a comparable amount of lithium on copper foil is vastly improved for Li2S counter electrode as opposed to Li-metal counter electrode. The exceptional robustness of the Li2S system points to an intrinsic stabilization of the lithium-deposition process in Li-S batteries. Detailed morphological and compositional characterization of the deposited lithium shows that the polysulfide intermediates formed help modulate the plating of lithium by forming deposits of Li2S and Li2S2, which facilitate the formation of a smooth planar morphology. A comparison of various electrolyte formulations in full-cell configuration reveals a sharp divergence in electrochemical performance, which is not observed in half-cell configuration. Emanating from the disparate lithium-deposition characteristics in different electrolytes, this helps provide insights into the critical solid-electrolyte interphase (SEI) components on the lithium surface that enable dynamic stabilization of lithium plating and stripping in Li-S batteries. This configuration portrays a promising approach to a realistic and effective implementation of the lithium-metal anode in Li-S batteries. It is significant that a considerable enhancement in gravimetric and volumetric energy density can be achieved from eliminating excess lithium, while still maintaining reasonable electrochemical performance. The lithium-limited nature of this configuration makes it an ideal template for generating novel insights into the dynamics of lithium degradation and SEI formation. This work opens up a new frontier in research in Li-S batteries, bringing them one step closer to reality. Figure: Schematic illustration of the deposited lithium on the copper foil from a Cu || Li2S full cell and the electrochemical performance of the Li || Li2S half cell compared with Cu || Li2S full cell. Figure 1

(Invited) In-Situ Investigation of the Role of Boric Acid during the Electrodeposition of Cobalt

As interconnect dimensions continue to decrease, copper damascene faces significant challenges1. As a result, alternative metals such as cobalt are actively being explored. Because the standard reduction potential for cobalt is more cathodic than that of hydrogen evolution, the solution pH is a critical consideration for the plating process. Boric acid is a common component in plating solutions used for cobalt electrodeposition. It is believed that boric acid acts as a buffering agent that helps to stabilize bath pH and to prevent formation of cobalt hydroxide precipitate on the substrate2-4. Typically, maximum buffering occurs when the pH equals the pKa for the weak acid. Thus, it is not well understood how an additive such as boric acid, with a pKa value of 9.275, can act as a buffer in typical cobalt electroplating baths6-9 which often have pH ranges of 2-5. The aim of this work is to provide an improved understanding of the role of boric acid on the electrodeposition of cobalt. It is important to first determine the effects of boric acid on surface pH during cobalt deposition. We present a technique for in-situ monitoring of surface pH as a function of applied plating current density. This is done using a tungsten microelectrode which can function as an indicator electrode for the hydrogen ion activity10-12. Results for plating cobalt from a virgin makeup solution (VMS) will be compared to those containing boric acid to demonstrate that this additive acts as a buffer at the electrode surface where pH values drastically increase during metal deposition. Electrochemical impedance spectroscopy (EIS) data will aid in understanding the surface adsorption behavior as well as the coulombic plating efficiency. In addition to surface specific information, potentiometric titration data will be presented to explain buffering effects in the bulk electrolyte. References R. Akolkar, "Current Status and Advances in Damascene Electrodeposition," Encyclopedia of Interfacial Chemistry, pp. 24-31, 2018.N. Zech and D. Landolt, "The influence of boric acid and sulfate ions on the hydrogen formation in Ni-Fe plating electrolytes," Electrochimica Acta, vol. 45, no. 21, pp. 3461-3471, 2000.J. P. Hoare, "On the Role of Boric Acid in the Watts Bath," Journal of The Electrochemical Society, vol. 133, no. 12, pp. 2491-2494, 1986.J. Santos, R. Matos, F. Trivinho-Strixino and E. C. Pereira, "Effect of temperature on Co electrodeposition in the presence of boric acid," Electrochimica Acta, vol. 53, pp. 644-649, 2007.D. R. Lide, CRC Handbook of Chemistry and Physics, Boca Raton, FL: CRC Press, 2005, p. 1274.M. A. Rigsby, L. J. Brogan, N. V. Doubina, Y. Liu, E. C. Opocensky, T. A. Spurlin, J. Zhou and J. D. Reid, "The Critical Role of pH Gradient Formation in Driving Superconformal Cobalt Deposition," Journal of The Electrochemical Society, vol. 166, no. 1, pp. D3167-D3174, 2019.Y. Hu and Q. Huang, "Effects of Dimethylglyoxime and Cyclohexane Dioxime on the Electrochemical Nucleation and Growth of Cobalt," Journal of The Electrochemical Society, vol. 166, no. 1, pp. D3175-D3181, 2019.C. H. Lee, J. E. Bonevich, J. E. Davies and T. P. Moffat, "Superconformal Electrodeposition of Co and Co–Fe Alloys Using 2-Mercapto-5-benzimidazolesulfonic Acid," Journal of The Electrochemical Society, vol. 156, no. 8, pp. D301-D309, 2009.D. Josell, M. Silva and T. Moffat, "Superconformal Bottom-Up Cobalt Deposition in High Aspect Ratio Through Silicon Vias," ECS Transactions, vol. 75, no. 2, pp. 25-30, 2016.S. E. S. E. Wakkad, H. A. Rizk and I. G. Ebaid, "The Electrochemical Behavior of the Tungsten Electrode and the Nature of the Different Oxides of the Metal," The Journal of Physical Chemistry, vol. 59, no. 10, pp. 1004-1008, 1955.J. O. Park, C.-H. Paik and H. C. Alkire, "Scanning Microsensors for Measurement of Local pH Distributions at the Microscale," Journal of The Electrochemical Society, vol. 8, no. 143, p. 174, 1996.E. Webb and R. Alkire, "Pit Initiation at Single Sulfide Inclusions in Stainless Steel," Journal of The Electrochemical Society, vol. 146, no. 6, p. B280, 2002.

Generation of Organoids from Mouse Extrahepatic Bile Ducts.

Cholangiopathies, which affect extrahepatic bile ducts (EHBDs), include biliary atresia, primary sclerosing cholangitis, and cholangiocarcinoma. They have no effective therapeutic options. Tools to study EHBD are very limited. Our purpose was to develop an organ-specific, versatile, adult stem cell-derived, preclinical cholangiocyte model that can be easily generated from wild type and genetically engineered mice. Thus, we report on the novel technique of developing an EHBD organoid (EHBDO) culture system from adult mouse EHBDs. The model is cost-efficient, able to be readily analyzed, and has multiple downstream applications. Specifically, we describe the methodology of mouse EHBD isolation and single cell dissociation, organoid culture initiation, propagation, and long-term maintenance and storage. This manuscript also describes EHBDO processing for immunohistochemistry, fluorescent microscopy, and mRNA abundance quantitation by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). This protocol has significant advantages in addition to producing EHBD-specific organoids. The use of a conditioned medium from L-WRN cells significantly reduces the cost of this model. The use of mouse EHBDs provides almost unlimited tissue for culture generation, unlike human tissue. Generated mouse EHBDOs contain a pure population of epithelial cells with markers of endodermal progenitor and differentiated biliary cells. Cultured organoids maintain homogenous morphology through multiple passages and can be recovered after a long-term storage period in liquid nitrogen. The model allows for the study of biliary progenitor cell proliferation, can be manipulated pharmacologically, and may be generated from genetically engineered mice. Future studies are needed to optimize culture conditions in order to increase plating efficiency, evaluate functional cell maturity, and direct cell differentiation. Development of co-culture models and a more biologically neutral extracellular matrix are also desirable.

A Simple and Useful Predictive Assay for Evaluating the Quality of Isolated Hepatocytes for Hepatocyte Transplantation

No optimal assay for assessing isolated hepatocytes before hepatocyte transplantation (HTx) has been established, therefore reliable and rapid assays are warranted. Isolated rat hepatocytes were dipped in a water bath (necrosis model), and were also cultured with Okadaic acid (apoptosis model) or vehicle, followed by cellular assessment including trypan blue exclusion (TBE) viability, ADP /ATP ratio, plating efficiency (PE), DNA quantity and ammonia elimination. Hepatocytes were transplanted into the liver of analbuminemic rats, subsequently engraftment was assessed by serum albumin and the histology of transplanted grafts. In the necrosis model, the ADP/ATP ratio was strongly and negatively correlated with the TBE (R2 = 0.559, P < 0.001). In the apoptosis model, the ADP/ATP ratio assay, PE, DNA quantification and an ammonia elimination test clearly distinguished the groups (P < 0.001, respectively). The ADP/ATP ratio, PE and DNA quantity were well-correlated and the ammonia elimination was slightly correlated with the transplant outcome. TBE could not distinguish the groups and was not correlated with the outcome. The ADP/ATP ratio assay predicted the transplant outcome. PE and DNA quantification may improve the accuracy of the retrospective (evaluations require several days) quality assessment of hepatocytes. The ADP/ATP ratio assay, alone or with a short-term metabolic assay could improve the efficiency of HTx.

Effect of temperature on ultrasound-assisted electroless nickel-boron plating

Electroless nickel-boron coatings have several advantages over electroplated nickel and electroless nickel-phosphorous: they are harder than both other coatings and, as all electroless coatings, can be applied easily to complex shapes and all substrates, even non conducting ones, contrary to electroplated coatings. Preliminary testing has proved that ultrasound assistance helps improve their properties by increasing the plating rate while conserving the properties of the coating.In this study, the effect of plating temperature on mechanically agitated and ultrasonic assisted electroless nickel-boron deposition was investigated: deposition was performed in two different configurations: one with a classical mechanical agitation at 300 rpm and the other employing ultrasound at a frequency of 35 kHz. In addition, different temperatures in the 80–95 °C range were tested. The increase of plating rate previously observed was confirmed and it was possible to lower slightly plating temperature while conserving plating efficiency, which decreases evaporation of the solution. Morphological, mechanical, corrosion and wear characterization were performed on the coatings, as well as tribocorrosion studies in an alkaline environment (0.1 M NaCl). Ultrasound-assisted coatings presented tribocorrosion behaviour that was similar or better than the standard ones.

Changes in the Proliferation Rate, Clonogenicity, and Radiosensitivity of Cultured Cells During and After Continuous Low-Dose-Rate Irradiation.

We investigated the effects of continuous low-dose radiation on proliferation, clonogenicity, radiosensitivity, and repair of DNA double-strand breaks (DSBs) in human salivary gland (HSG) tumor cells. Human salivary gland cells were cultured on acrylic boards above very-low-dose (4.3 μSv/h) or low-dose (27 μSv/h) radiation-emitting sheets or without sheets. Total cell numbers and plating efficiencies were compared among the 3 groups every 1 or 2 weeks until 6 weeks after starting culture. At 2, 4, and 6 weeks, surviving fractions of HSG cells after irradiation at 2 to 8 Gy cultured on the very-low-dose or low-dose sheets were compared to those of the control. At 4 weeks, HSG cells irradiated at 2 Gy were assessed for phosphorylated histone (γH2AX) foci formation, and DSBs were evaluated. No significant differences were observed in total cell number or plating efficiencies with or without low-dose-emitting sheets. The surviving fractions after irradiation of the very-low-dose group at 2 to 6 weeks and those of the low-dose group at 2 to 4 weeks were higher than those of the control (P < .01). Thus, a radioadaptive response was clearly demonstrated. From the γH2AX foci quantification, the adaptive responses were considered to be associated with the efficient repair of DSB, especially slow repair, in this cell line.

The putative tumour suppressor protein Latexin is secreted by prostate luminal cells and is downregulated in malignancy

Loss of latexin (LXN) expression negatively correlates with the prognosis of several human cancers. Despite association with numerous processes including haematopoietic stem cell (HSC) fate, inflammation and tumour suppression, a clearly defined biological role for LXN is still lacking. Therefore, we sought to understand LXN expression and function in the normal and malignant prostate to assess its potential as a therapeutic target. Our data demonstrate that LXN is highly expressed in normal prostate luminal cells but downregulated in high Gleason grade cancers. LXN protein is both cytosolic and secreted by prostate cells and expression is directly and potently upregulated by all-trans retinoic acid (atRA). Whilst overexpression of LXN in prostate epithelial basal cells did not affect cell fate, LXN overexpression in the luminal cancer line LNCaP reduced plating efficiency. Transcriptome analysis revealed that LXN overexpression had no direct effects on gene expression but had significant indirect effects on important genes involved in both retinoid metabolism and IFN-associated inflammatory responses. These data highlight a potential role for LXN in retinoid signaling and inflammatory pathways. Investigating the effects of LXN on immune cell function in the tumour microenvironment (TME) may reveal how observed intratumoural loss of LXN affects the prognosis of many adenocarcinomas.

Targeting of BRM Sensitizes BRG1-Mutant Lung Cancer Cell Lines to Radiotherapy.

Targeting of epigenetic regulators as the chromatin remodeler SWI/SNF is proving to be a promising therapeutic strategy for individualized treatment of cancer patients. Here, we tested whether targeting one of the two mutually exclusive subdomains of the SWI/SNF complex BRM/SMARCA2 can sensitize specifically non-small cell lung carcinoma (NSCLC) cells with mutations in the other subunit BRG1/SMARCA4 toward ionizing radiation (IR). Knockdown of BRM with siRNA or shRNA and its consequences for radiation sensitivity as measured by clonogenic survival and plaque-monolayer control was studied in different NSCLC lines with or without BRG1 mutations and in primary fibroblasts. Furthermore, the effect on double-strand break (DSB) repair markers measured by immunofluorescence staining of 53BP1-, γ-H2AX-, and Rad51-foci was investigated. BRG1-mutated cell lines showed an increased surviving fraction compared with BRG1 proficient cells. Depletion of BRM (i) leads to a decreased proliferation rate and plating efficiency specifically in BRG1-mutated cells, (ii) specifically sensitized BRG1-mutant NSCLC cells toward IR as characterized by a survival reducing factor of 0.63 [95% confidence interval (CI), 0.57-0.69] in the dose range between 2 and 6 Gy, and (iii) decreased the tumor control doses after daily fractionation at 4 Gy in BRG1-mutant NSCLC cell lines A549 and H1299 in minimonolayers by 9.9% ± 1.3% and 13.6% ± 1.8%, respectively. In addition, an increase of residual Rad51-foci at 24 hours after irradiation in BRG1-mutant cells was demonstrated. Therefore, targeting of BRM in combination with radiotherapy is supposed to improve the therapeutic outcome of lung cancer patients harboring BRG1 mutations.The present study shows that the moderate radioresponsiveness of NSCLC cells with BRG1 mutations can be increased upon BRM depletion that is associated with a prolonged Rad51-foci prevalence at DNA DSBs.

Application of the salt stress to the protoplast cultures of the carrot (Daucus carota L.) and evaluation of the response of regenerants to soil salinity

This is the first study to generate carrot plants for enhanced salinity tolerance using a single-cell in vitro system. Protoplasts of three carrot accessions were exposed to treatment by seven different concentrations of NaCl (10–400 mM). Salt concentrations higher than 50 mM decreased plating efficiency and those of 200–400 mM of NaCl completely arrested mitotic divisions of cultured cells. The protoplast-derived plants from the control and 50–100 mM NaCl treatment were subjected to an 8-week salt stress in greenhouse conditions induced by salinized soil (EC 3 and 6 mS cm−1). 50 mM NaCl stress applied in vitro induced polyploidy among regenerated plants. The regenerants obtained from the 50 and 100 mM NaCl-treated protoplast cultures grown in saline soil had a higher survival rate compared to the regenerants from the control cultures. The salt-stressed plants accumulated anthocyanins in petioles and produced denser hairs on leaves and petioles in comparison to the control plants. Salt stress influenced pollen viability and seed setting of obtained regenerants. The results suggest that salt stress applied in vitro in protoplast cultures creates variation which allows alleviating the negative effects of salt stress on the development and reproduction of the carrot.

&lt;p&gt;Establishment and characterization of the GC-030-35 cell line derived from gastric hepatoid adenocarcinoma&lt;/p&gt;

PurposeGastric hepatoid adenocarcinoma is a rare subtype of primary gastric cancer and is a high-grade form of malignancy. However, the pathogenesis and molecular biology of gastric hepatoid adenocarcinoma remain poorly understood. The aim of this study was to establish and characterize a new human gastric hepatoid adenocarcinoma cell line, GC-030-35.Materials and methodsThe GC-030-35 cell line was established from tumor cells from a 58-year-old Chinese man with gastric hepatoid adenocarcinoma. The cultured cells underwent immunocytochemistry and flow cytometry to confirm the tumor cell phenotype. RNA sequencing was performed to analyze the differences in gene expression between GC-030-35 cells compared with normal gastric epithelial cells. A zebrafish assay was performed. Gene enrichment analysis and interrogation of the bioinformatics databases, the Gene Ontology (GO) database and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, were used for pathway analysis.ResultsFlow cytometry analysis of the GC-030-35 cells showed a positive expression rate for CD44+ of 10.7%, high cell clonality, an average plating efficiency of 32%, cell-doubling time of 29.2 hours, and cell proliferation for >15 generations in serial culture. The zebrafish assay showed the ability of the GC-030-35 cells to proliferate, promote angiogenesis, and metastasize. RNA sequencing identified the functional clustering of 6,601 differentially expressed genes of GC-030-35, which were significantly different when compared with nonneoplastic gastric epithelial cells. Pathway enrichment analysis and interrogation of the GO and KEGG bioinformatics databases identified genes for microbial metabolism in diverse environments (63 genes), metabolism of xenobiotics by cytochrome P450 (CYP450; 25 genes), and the drug metabolism cytochrome P450 (28 genes).ConclusionA human gastric hepatoid adenocarcinoma cell line, GC-030-35, was developed and characterized by comparison with normal gastric epithelial cells. Bioinformatics and gene analysis data showed that the CYP450 gene was significantly differentially expressed by GC-030-35 cells.

(Invited) Exploring the Impact of Mechanical Pressure on the Performance of Anode-Free Lithium Metal Cells

Research on lithium metal batteries has made a resurgence in the quest to surpass the energy density of conventional lithium ion batteries. Many reports in the literature utilize Li-metal cells with significant excess lithium, thereby artificially inflating the cycling efficiency and cycle life of such cells since lost lithium inventory can be replaced from a vast reservoir. However, utilizing significant excess lithium can drastically reduce the energy density of Li-metal cells even below that of conventional Li-ion as shown in Figure 1; at 200% excess lithium, the theoretical volumetric capacity of Li of 2060 mAh/L becomes 687 mAh/L, lower than the volumetric capacity of graphite of 719 mAh/L. Clearly cells with significant excess lithium are not poised to succeed conventional lithium ion cells in delivering superior energy density. In contrast, anode-free cells do not utilize excess lithium and thus can deliver the maximum theoretical capacity of lithium metal. In an anode-free cell, the lithium ions stored in the positive electrode plate on a bare current collector during the first charge to form a lithium metal anode in-situ.1,2 As such, lithium metal is not required during the construction of anode-free cells, resulting in a cheaper, safer and more practical Li-metal cell. In this work, we evaluate anode-free Li-metal pouch cells (NMC532||Cu) with operando pressure measurements constrained to different stack pressures between 75-2205 kPa with two different electrolytes, 1M LiPF6 fluoroethylene carbonate: diethyl carbonate (FEC:DEC 1:2) and 1M LiPF6 fluoroethylene carbonate:bis(2,2,2-trifluoroethyl) carbonate (FEC:TFEC 1:2). Increasing the initial average pressure from 75 to 2205 kPa was found to generally improve cycling performance, with the most significant benefits achieved up to 1205 kPa. Cells containing FEC:TFEC electrolyte exhibited a superior initial plating efficiency than FEC:DEC cells. The benefit of constraining cells containing FEC:TFEC electrolyte is shown in Figure 1. Although generally beneficial, we found that the effect of increased pressure on the performance of cells with different solvent systems was not equal, particularly at high pressures, indicating that the physical properties of the electrolyte play an important role in cells constrained to higher pressures between 1205 and 2205 kPa. This work presents anode-free cells which deliver a larger energy density than conventional Li-ion cells for 50 cycles. Figure 1

Lithium dendrite-free plating/stripping: a new synergistic lithium ion solvation structure effect for reliable lithium-sulfur full batteries.

A new synergistic lithium ion solvation structure effect tailored by mixed organic/inorganic lithium salts is introduced to mitigate the growth of lithium-dendrites. A uniform lithium plating, lower polarization and higher coulombic efficiency are confirmed by lithium plating/stripping. The effect of different solvation structures was analyzed by Raman spectroscopy and simulations. Then, a reliable lithium-sulfur full battery with high performance over hundreds of cycles was configured.

Umbilical Cord Tissue Derived Mesenchymal Stem Cells can Differentiate into Skin Cells.

Autologous skin grafts are used to treat severe burn wounds, however, the availability of adequate donor sites makes this option less practical. Recently, stem cells have been used successfully in tissue engineering and in regenerative medicine. The current study aims to differentiate umbilical cord tissue derived mesenchymal stem cells (CT-MSCs) into skin cells (fibroblasts and keratinocytes) for use to treat severe burn wounds. After isolation, MSCs were characterized and their growth characteristics were determined. The cells were induced to differentiate into fibroblasts and keratinocytes using respective induction medium. Results indicated that CT-MSCs were spindle shaped, plastic adherent and positive for CD29, CD44, CD73, CD90 markers. CT-MSCs also showed high proliferative potential as indicated by cumulative population doubling, doubling time and plating efficiency. The MSCs were successfully differentiated into fibroblast and keratinocytes as indicated by morphological changes and expression of lineage specific genes. We propose that these differentiated skin cells which are derived from CT-MSCs can thus be used for the development of bioengineered skin; however, further studies are required to evaluate the utility of these substitutes.