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Purpose The clonogenic property and radiobiological responses of a fish brain endothelial cell line, eelB, derived from the American eel were studied. Methods Clonogenic assays were performed to determine the plating efficiency of the eelB cells and to evaluate the clonogenic survival fractions after direct irradiation to low-dose low-LET gamma radiation or receiving irradiated cell conditioned medium in the bystander effect experiments. Result eelB had the second highest plating efficiency ever reported to date for fish cell lines. Large eelB macroscopic colonies could be formed in a short period of time and were easy to identify and count. Unlike with other fish clonogenic cell lines, which had a relatively slow proliferation profile, clonogenic assays with the eelB cells could be completed as early as 12 days in culture. After direct irradiation with gamma rays at low doses ranging from 0.1 Gy to 5 Gy, the dose-clonogenic survival curve of the eelB cell line showed a linear trend and did not develop a shoulder region. A classical radio-adaptive response was not induced with the clonogenic survival endpoint when the priming dose (0.1 or 0.5 Gy) was delivered 6 h before the challenge dose (3 or 5 Gy). However, a radio-adaptive response was observed in progeny cells that survived 5 Gy and developed lethal mutations. eelB appeared to lack the ability to produce damaging radiation-induced bystander signals on both eelB and HaCaT recipient cells. Conclusion eelB cell line could be a very useful cell model in the study of radiation impacts on the aquatic health.

Background and purpose The SCF/c-Kit pathway is often overexpressed in human tumors leading to an enhanced tumorigenesis, proliferation and migration. It was now tested for NSCLC and prostate cancer cells growing in 2D and 3D whether the inhibition of this pathway can be used to achieve a significant radiosensitization and whether a respective biomarker may be identified. Material and methods Experiments were performed with different cancer cell lines (NSCLC: H23, H520, H226, H1975 and PrCa: DU145) growing either under 2D or 3D conditions. Expression of SCF and c-Kit was determined by RT-PCR and Western blot, SCF was knocked down by siRNA, c-Kit was inhibited by ISCK03 inhibitor and cell survival was determined by colony formation assay. Results There is a profound variation in the expression of both c-Kit and SCF with no association between each other. Neither levels did correlate with the respective cellular radiosensitivity determined for 2D or 3D with only a trend seen for SCF. Knock-down of SCF was generally found to result in no or only minor reduction of plating efficiency or cellular radioresistance. A significant reduction was only obtained for H520 cells characterized by an extreme over-expression of SCF. The inhibition of c-Kit by a specific inhibitor was also found to result only in minor radiosensitization. Conclusion Generally, the SCF/c-Kit pathway does not have a dominant effect on both, cell survival and radioresponse and, as a consequence, knockdown of this pathway does not result in a strong effect on radioresistance, except when SCF is strongly over-expressed.

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.

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.

Newly expanded in vitro leaves of Actinidia eriantha were used for protoplast isolation. Protoplasts were cultured in liquid MS medium (lacking NH4NO3) supplemented with 2,4-D (2,4-dichlorophenoxyacetic acid) and 0.4 M glucose. The plating efficiency after 3 weeks of culture was 19.4%, and calli were recovered without addition of fresh medium. These calli regenerated shoots on transfer to MS medium containing 2.28 μM zeatin and 0.57 μM IAA (indole-3-acetic acid). Regenerated shoots were rooted by immersion in 20 ppm IBA (indole-3-butyric acid) solution before culturing on half-strength MS medium lacking growth regulators. Somaclonal variation, in terms of chromosome number and nuclei per cell of protoplast-derived plants, was estimated.

Novel approaches, involving supplementation of aqueous culture medium with haemoglobin solution (Erythrogen), in the presence or absence of the copolymer surfactant, Pluronic F-68, have been evaluated to facilitate cellular oxygen availability to promote mitotic division. Cell-suspension-derived protoplasts of albino Petunia hybrida cv. Comanche were cultured for up to 45 days in KM8P medium containing 1:50–1:500 (vol:vol) Erythrogen. The mean initial protoplast plating efficiency after 9 days with 1:50 Erythrogen (18.5%) was significantly greater (P

‘Anode-free’ batteries present a significant advantage due to their substantially higher energy density and ease of assembly in a dry air atmosphere. However, potential safety issues due to lithium (Li) dendrite growth rather than smooth Li metal plating on the Cu current collector, and low cycling coulombic efficiency during their operation are delaying their practical implementation. To understand the interplay between Cu surface chemistry and the morphology of the plated Li, we studied the SEI formation on Cu and the preferential plating of Li using ssNMR, insitu NMR, XPS, ToF-SIMS, SEM and EIS. A native interphase layer (N-SEI) is formed instantaneously on copper current collectors upon their immersion in LiPF6-based electrolyte. The nature of the N-SEI is affected by the copper interface composition, homogeneity and formation time. In addition to the typical SEI components, it contains CuxO and its reaction products. Parasitic semi-reversible electrochemical reactions were observed with in-situ NMR measurements of Li plating efficiency during the first five cycles. The morphology of the plated lithium is affected by the SEI homogeneity, current density and rest time in the electrolyte before the plating. The preferential plating of Li is governed by the distribution of ionic conducting compounds rather than electronic conducting compounds.[i] [i] Menkin, Svetlana; O'Keefe, Christopher A.; Gunnarsdottir, Anna B.; Dey, Sunita; Pesci, Federico; Shen, Zonghao; et al. (2020): Towards an Understanding of the SEI Formation and Lithium Preferential Plating on Copper. ChemRxiv. Preprint. https://doi.org/10.26434/chemrxiv.12839792.v1

This work targets the identification of optimal rate-enhancement technique for the electroless fabrication of silver–ceramic porous membranes. The membranes were fabricated with porous ceramic supports of 4.5 µm and porosity of 19.3% and with alternate electroless plating (ELP) processes including conventional (CEP), sonication (SOEP), and surfactant (SIEP) plating baths. The alternate processes were evaluated based on the process (conversion, plating efficiency (η), average plating rate) and membrane characteristics (percent pore densification (PPD), metal loading index (MLI)), and their combinations (η/(MLI.PPD)). Among alternate processes, SOEP has been evaluated to be the best to yield silver–ceramic membranes.

The present work was focused on the preliminary stage of palladium composite membrane preparation through electroless plating and the effects of bath hydrodynamic properties on the deposited layer morphology. Eight samples of palladium composite membrane were prepared which were different in terms of plating time, solution stirring rate and bath agitating mode. Scanning electron microscopy and energy-dispersive X-ray spectroscopy were used for characterization of deposited layer. In comparison with the support rotating mode, the support fixing and the magnet rotating mode, which are caused running the solution toward the support, were led to a deposited layer with better quality, larger grains of deposited layer, more compacted plating and higher plating efficiency.

Devices that combine magnetic resonance imaging with linear accelerators (MRL) represent a novel tool for MR-guided radiotherapy. However, whether magnetic fields (MFs) generated by these devices affect the radiosensitivity of tumors is unknown. We investigated the influence of a 1.5-T MF on cell viability and radioresponse of human solid tumors. Human head/neck cancer and lung cancer cells were exposed to single or fractionated 6-MV X-ray radiation; effects of the MF on cell viability were determined by cell plating efficiency and on radioresponsiveness by clonogenic cell survival. Doses needed to reduce the fraction of surviving cells to 37% of the initial value (D0s) were calculated for multiple exposures to MF and radiation. Results were analyzed using Student's t-tests. Cell viability was no different after single or multiple exposures to MRL than after exposure to a conventional linear accelerator (Linac, without MR-generated MF) in 12 of 15 experiments (all P > 0.05). Single or multiple exposures to MF had no influence on cell radioresponse (all P > 0.05). Cells treated up to four times with an MRL or a Linac further showed no changes in D0s with MF versus without MF (all P > 0.05). In conclusion, MF within the MRL does not seem to affect in vitro tumor radioresponsiveness as compared with a conventional Linac. Bioelectromagnetics. 37:471-480, 2016. © 2016 Wiley Periodicals, Inc.

Al-doped HfO2 nanoparticles were prepared at room-temperature using a novel one-step, straightforward, high-yield, efficient and green process. It includes a mechanically activated metathesis reaction that promotes the direct crystallization of cubic HfO2 (c-HfO2) in milling. Al-cation was used as dopant, achieving c-HfO2 stabilization at low content (5% mol). The c-HfO2 is stable up to 380 °C without exhibiting transformation from the cubic to monoclinic polymorph. The c-HfO2 showed spherical-like morphology with diameter below 7 nm. Human Prostate Epithelial Cells (HPrEC) were used to evaluate nanoparticles toxicity. Cells viability and survival were determined performing MTS and colony formation assays. Nanoparticles were found harmless for cells metabolic activity at concentrations up to 1000 μg/mL. A decrease of ~30% on HPrEC plating efficiency was observed at the highest concentration.

Human pluripotent stem cells (hPSCs) are viewed as promising candidates for applications in regenerative medicine and therapy due to their proliferative and pluripotent properties. However, obtaining clinically significant numbers of hPSCs remains a limiting factor and impedes their use in therapeutic applications. Conventionally, hPSCs are cultured on two-dimensional surfaces coated with a suitable substrate, such as Matrigel™. This method, however, requires a large surface area to generate sufficient cell numbers to meet clinical needs and is therefore impractical as a manufacturing platform for cell expansion. In addition, the use of enzymes for cell detachment and small molecule inhibitors to increase plating efficiency may impact future cell behavior when used for routine subculturing. In this study, we describe a protocol to generate and maintain hPSC aggregates in a three-dimensional suspension culture by utilizing thermoresponsive nanobridges. The property of the polymer used in the nanobridges enables passaging and expansion through a temperature change in combination with mechanically applied shear to dissociate aggregates; thus, we eliminate the need of enzymes or small molecules for cell dissociation and viability, respectively. Utilizing this platform, maintenance of human embryonic stem cells for three continuous passages demonstrated high expression levels in key pluripotent markers.

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 P

An efficient protocol for plant regeneration was developed from protoplasts of Gentiana macrophylla Pall. through somatic embryogenesis. Viable protoplasts were isolated from cell suspensions derived from young seedling leaves in an enzyme solution containing 2 % Cellulase Onozuka R-10, 0.5 % Macerozyme R-10, 0.5 % Hemicellulase, and 0.4 M sorbitol with a yield of 6.2 × 106 protoplasts g−1 fresh weight. Liquid, solid–liquid double layer (sLD) and agar-pool (aPL) culture systems were used for protoplast culture. The frequency of protoplast cell divisions and colony formations in aPL culture were significant (p < 0.05) higher than those in liquid and sLD cultures. The highest division frequency and plating efficiency were 37.7 % and 16.5 %, respectively, in aPL culture supplemented with 2 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-d) and 0.5 mg l−1 6-benzylaminopurine (BA). Protoplast-derived microcalli obtained from aPL culture system were transferred to solid MS medium with a reduced concentration of 2,4-d (0.5 mg l−1) to promote the formation of embryogenic calli. Somatic embryos developed into plantlets on MS medium supplemented with 2 mg l−1 BA at a rate of 51.3 %. RAPD analysis of G. macrophylla revealed a low variation among regenerants.

This article presents our further in vitro studies into the morphogenic potential of gentian cells, organs, and tissues after modification of their genome. The objective was to study the effect of electroporation and the introduction of foreign genes on the morphogenic potential of Gentiana kurroo embryogenic cell suspension protoplasts. Protoplasts were electroporated with DNA plasmids carrying nptII and bar genes. The stability of cell membranes, the contents of electroporation buffer, the length of electric pulse, the number of pulses and the strength of the electric field were studied. We determined the highest electroporation efficiency by evaluating the highest protoplast survival rate under specific physical conditions. The best results were achieved in the presence of EB1 electroporation buffer where the viability of protoplasts was 70.1%. Protoplast survival at this higher level required culture temperatures near 0EC, and a 20 μs electric pulse with an electric field of 1.0 kV/cm. After seven days of agarose embedded protoplast culture, a selective agent – kanamycin – was introduced to the medium. The cell transformation effect was improved by a long term culture of callus, regenerated somatic embryos and transformants in the presence of 50 mg/l kanamycin.

rad52-ΔΔ and, to a lesser extent, rad51-ΔΔ deletants of Candidaalbicans displayed slow growth and aberrant filamentous morphology whereas rad59-ΔΔ mutants, both by growth rate and morphology resembled wild type. In this study, we have constructed pair-wise double deletants to analyze genetic interactions among these homologous recombination (HR) proteins that affect growth and morphology traits. When grown in liquid YPD medium, double mutant rad51-ΔΔ rad59-ΔΔ exhibited growth rates, cell and colony morphologies, and plating efficiencies that were not significantly different from those observed for rad51-ΔΔ. The same was true for rad52-ΔΔ rad59-ΔΔ compared to rad52-ΔΔ. Slow growth and decreased plating efficiency were caused, at least in part, by a decreased viability, as deduced from FUN1 staining. Flow cytometry and microscopic studies of filamentous mutant populations revealed major changes in cell ploidy, size and morphology, whereas DAPI staining identified complex nuclear rearrangements in yeast and filamentous cells. These phenotypes were not observed in the rad59-ΔΔ mutant populations. Our results show that abolishing Rad51 functions induces the appearance of a subpopulation of aberrant yeast and filamentous forms with increased cell size and ploidy. The size of this complex subpopulation was exacerbated in rad52-ΔΔ mutants. The combination of filamentous cell morphology and viability phenotypes was reflected on the colony morphology of the respective mutants. We conclude that the rad52 mutation is epistatic to rad51 for all the morphological traits analyzed. We discuss these results in the light of the several functions of these recombination genes.

Objective Human amniotic epithelial cells (hAECs) which are isolated from the amniotic membrane have stem cell-like properties and immunomodulatory effects. Several protocols have been proposed for isolation of hAECs, nevertheless, there is no report concerning isolation of highly viable hAECs, with desirable yield, and without significant purity reduction. In the current study, a detailed protocol with some modification of previous ones is presented in which the amendments led to isolation of hAECs with high purity, yield and viability. Moreover, isolated hAECs were subjected to immuno-phenotyping and their physiological status was assessed using a proliferation assay. Results The average yield of obtained hAECs using the new modified method was 190 × 106 cells with a mean viability of 87%, with less than 1% contamination with mesenchymal stem cells (MSCs). The isolated cells were > 95% positive for the epithelial cell markers. The lowest initial plating efficiency of the cells was 80%. Freshly isolated hAECs had the ability to proliferate for 5–6 passages in a standard culture medium.

Thermal damage rate processes in biological tissues are usually characterized by a kinetics approach. This stems from experimental data that show how the transformation of a specified biological property of cells or biomolecule (plating efficiency for viability, change in birefringence, tensile strength, etc.) is dependent upon both time and temperature. Here, two disparate approaches were used to study thermal damage rate processes in cultured retinal pigment epithelial cells. Laser exposure (photothermal) parameters included 2-μm laser exposure of non-pigmented cells and 532-nm exposures of cells possessing a variety of melanosome particle densities. Photothermal experiments used a mid-IR camera to record temperature histories with spatial resolution of about 8 μm, while fluorescence microscopy of the cell monolayers identified threshold damage at the boundary between live and dead cells. Photothermal exposure durations ranged from 0.05-20 s, and the effects of varying ambient temperature were investigated. Temperature during heat transfer using a water-jacketed cuvette was recorded with a fast microthermister, while damage and viability of the suspended cells were determined as percentages. Exposure durations for the heat transfer experiments ranged from 50- 60 s. Empirically-determined kinetic parameters for the two heating methods were compared with each other, and with values found in the literature.

Effect of high-pressure torsion (HPT) deformation on biocompatibility and surface chemistry of TiNi was systematically investigated. Ti–50 mol% Ni was subjected to HPT straining for different numbers of turns, N = 0.25, 0.5, 1, 5 and 10 at a rotation speed of 1 rpm. X-ray photoelectron spectroscopy observations after 7 days of cell culture revealed the changes in the surface oxide composition, enrichment of Ti and detection of nitrogen derived from organic molecules in the culture medium. Plating efficiency of L929 cells was slightly increased by HPT deformation though no significant difference was observed. Albumin adsorption was higher in HPT-deformed samples, while vitronectin adsorption was peaked at N = 1. HPT deformation was also found to effectively suppress the Ni ion release from the TiNi samples into the cell culture medium even after the low degree of deformation at N = 0.25.

Elaborating upon the role of surfactant and sonication, this work addresses the combinatorial plating characteristics for the fabrication of nickel ceramic composite membranes. Specific novelty corresponds to controlled addition of hydrazine hydrate as a reducing agent to nickel electroless plating bath. The experimental investigations evaluated the optimality of solution concentrations of cetyltrimethylammonium bromide (CTAB) and sodium dodecyl sulphate surfactants during coupled sonication and surfactant based electroless plating. Among both surfactants, CTAB surfactant with a solution concentration of 4 CMC provided maximum combinations of pore densification (73·4%), metal film thickness (8·4 μm) and plating efficiency (89·9%). In all, it was observed that with CTAB surfactant the pore densification and plating efficiencies increased with increasing surfactant solution concentration from 1–4 CMC and were maximum at 4 CMC which drastically decreased at 6 CMC respectively probably due to surfacta...

This work addresses the effect of palladium solution concentration on combinatorial plating characteristics of surfactant and sonication coupled electroless plating baths for the fabrication of dense palladium films on porous stainless steel substrates. All plating experiments were carried out using palladium solution concentration of 0·005 and 0·01M plating baths with cetyltrimethylammonium bromide surfactant at four critical micelle concentrations and loading ratio of 203 cm2 L−1. The evaluated combinatorial plating characteristics include selective conversion, plating efficiency, plating rate, Pd film thickness and per cent pore densification. The enhancement in palladium solution concentration was found to be insignificant to provide better combinatorial plating characteristics, and the plating bath consisting of 0·005M Pd solution concentration has provided 99·98% per cent pore densification with a Pd film thickness of 8·81 μm and plating efficiency of 80·69%.

Background aims Cord tissue (CT) storage is promoted as an opportunity to preserve a source of mesenchymal stromal cells (MSCs) for future use. We analyzed maximal MSC yields from fresh and frozen CT including functional capacity after long-term cryopreservation as a means of assessing potential utility. Methods CT was evaluated immediately upon harvest or frozen and banked for 5 years before analysis. Upon thawing, cell viability and yield were determined, as were growth characteristics and the ability to differentiate into various tissues. After thawing, enzymatic digestion of CT to release MSCs resulted in poor cell recoveries and few viable cells, requiring explant cultures to recover sufficient cell numbers for analysis. Upon expansion of surviving cells, fluorescence-activated cell sorter analysis showed the cells to be MSCs based on phenotype (CD34–, 45–, 44+, 90+, 105+) and function (ability to form adipocytes and osteoblasts). Frozen CT, however, exhibited decreased plating efficiency, increased doubling times but near equivalent maximum cell expansion, compared with fresh CT. Conclusions Poor cell yields and recoveries, along with slower growth characteristics, make frozen CT a less-than-optimal choice for MSC banking, despite good functional recovery. In addition, because the amount of fresh CT available at birth is limited and total MSC yields are low, even fresh CT-MSC requires extensive in vitro expansion before clinical use, which limits it application.

In the present study, the role of monoglucosyl-rutin as a potential radioprotector was investigated using mammalian cell culture models. Cell survival and DNA damage were assessed using colony formation, sister chromatid exchange and γH2AX assays. It was demonstrated that monoglucosyl-rutin was able to increase cell survival when exposed to ionizing radiation, possibly by decreasing the amount of base damage experienced by the cell. However, the present study also demonstrated that, despite monoglucosyl-rutin exhibiting radioprotective effects at low doses, high doses of monoglucosyl-rutin led to a decrease in plating efficiency and an increased doubling time. This effect may be due to double-strand breaks caused by high concentrations of monoglucosyl-rutin.

Three different regeneration systems, viz. regeneration through callus cultures using embryonic explant, direct regeneration using shoot bud/nodal segments as explant and regeneration through cell suspension culture using cotyledonary explant (for the induction of transgenic callus for suspension culture) were evaluated to see their effect on transfer of Cry1A(b) gene to Punica granatum L. cv. Kandhari Kabuli through Agrobacterium mediated transformation. Pre-conditioning and co-cultivation durations had a marked effect on transformation frequency of different explants. Out of different explants used (embryo, shoot bud, and cotyledon) for different regeneration systems cotyledonary explant showed highest putative transformation frequency (13.54%) inducing callus on selective medium for carrying out cell suspension culture to regenerate transgenic shoots. Despite of the highest transformation frequency obtained from the cotyledon explant, the plating efficiency of the transgenic cells generated through the transgenic callus (callus formed from the cotyledonary explant) during cell suspension culture was found to be very low (0.7%). Thus the plating efficiency has also played worth mentioning role in the regeneration of transformants following cell suspension culture. Among the three regeneration systems, regeneration through callus cultures using embryonic explant was found to be best for regeneration of transformants. The highest per cent regeneration of 23.33 was obtained from the putative transgenic embrogenic calli. Successful genetic transformation in the transformed plantlets was confirmed by PCR analysis. The transformation system thus developed is valuable and may be used to produce insect resistant trees.

This article presents the effects of the concentration and loading ratios of a cationic surfactant (CTAB) on the performance characteristics of sonication-induced electroless plating baths for the fabrication of dense palladium composite membranes. The plating experiments were conducted with dropwise addition of reducing agent at loading ratios of 203 and 407 cm2/L, a palladium solution concentration of 0.005 M, and surfactant solution concentrations of 1–4 CMC. A plating bath consisting of 0.005 M Pd with a surfactant concentration of 2 CMC at a loading ratio of 203 cm2/L provided a 100% dense palladium composite membrane after 8 h of sequential plating with a Pd film thickness of 11.16 μm and a plating efficiency of 96%. The identified process can provide dense Pd/PSS composite membranes at a fabrication cost of 13.27 $/cm2, which is 60% more cost-effective than the best-known SIEP process.

A new epithelial cell line, WAF was developed from caudal fin of freshwater shark, Wallago attu. The cell line was optimally maintained at 28 °C in Leibovitz-15 (L-15) medium supplemented with 20 % fetal bovine serum. The cell line was characterized by various cytogenetic and molecular markers. The cytogenetic analysis revealed a diploid count of 86 chromosomes at different passages. The origin of the cell lines was confirmed by the amplification of 547 and 654 bp sequences of 16S rRNA and cytochrome oxidase subunit I genes of mitochondrial DNA, respectively. WAF cells were characterized for their growth characteristics at different temperature and serum concentration. Epithelial morphology of the cell line was confirmed using immunocytochemistry. Further cell plating efficiency, transfection efficiency and viability of cryopreserved WAF cells was also determined. Cytotoxicity and genotoxicity assessment of cadmium salts on WAF cells by MTT, NR and comet assay illustrated the utility of this cell line as an in vitro model for aquatic toxicological studies. The cell line will be further useful for studying oxidative stress markers against aquatic pollutants.

The isolation and culture of suspension-derived protoplasts from two sugar beet (Beta vulgaris L.) genotypes are described. Immobilization of protoplasts in agarose resulted in high frequency divisions and microcallus regeneration, with plating efficiency (PE) being clearly genotype-dependent. In further studies towards asymmetric fusion experiments, the effect of different doses of ultraviolet radiation (UV) and iodoacetic acid (IA) on protoplast physiology was assessed. Viability of both treated (UV, IA) and untreated protoplasts (control) was determined by FDA staining, and the biological effect was evaluated by testing the ability of protoplasts to divide and to form calli. The results are discussed in terms of the applicability of the methods for the production of asymmetric protoplasts suitable for somatic hybridization within the genus Beta.

When deprived of FtsZ, Escherichia coli cells (VIP205) grown in liquid form long nonseptated filaments due to their inability to assemble an FtsZ ring and their failure to recruit subsequent divisome components. These filaments fail to produce colonies on solid medium, in which synthesis of FtsZ is induced, upon being diluted by a factor greater than 4. However, once the initial FtsZ levels are recovered in liquid culture, they resume division, and their plating efficiency returns to normal. The potential septation sites generated in the FtsZ-deprived filaments are not annihilated, and once sufficient FtsZ is accumulated, they all become active and divide to produce cells of normal length. FtsZ-deprived cells accumulate defects in their physiology, including an abnormally high number of unsegregated nucleoids that may result from the misplacement of FtsK. Their membrane integrity becomes compromised and the amount of membrane proteins, such as FtsK and ZipA, increases. FtsZ-deprived cells also show an altered expression pattern, namely, transcription of several genes responding to DNA damage increases, whereas transcription of some ribosomal or global transcriptional regulators decreases. We propose that the changes caused by the depletion of FtsZ, besides stopping division, weaken the cell, diminishing its resiliency to minor challenges, such as dilution stress., IMPORTANCE Our results suggest a role for FtsZ, in addition to its already known effect in the constriction of E. coli, in protecting the nondividing cells against minor stress. This protection can even be exerted when an inactive FtsZ is produced, but it is lost when the protein is altogether absent. These results have implications in fields like synthetic biology or antimicrobial discovery. The construction of synthetic divisomes in the test tube may need to preserve unsuspected roles, such as this newly found FtsZ property, to guarantee the stability of artificial containers. Whereas the effects on viability caused by inhibiting the activity of FtsZ may be partly overcome by filamentation, the absence of FtsZ is not tolerated by E. coli, an observation that may help in the design of effective antimicrobial compounds.

Bromelain is dotted with anticancer properties on various cancer cell lines. Anticancer pathways of bromelain, as well related intervening signalization are under investigation. Investigating the inhibitory potential of bromelain on AGS, PC3, and MCF7 cells proliferation and colony formation. The bromelain inhibitory potential on AGS, PC3, and MCF7 cells proliferation at various bromelain concentrations was assessed by MTT; thereby, bromelain potency on colony formation impediment was evaluated using clonogenic assays at determined 50% inhibitory concentrations (IC50) on four different cell densities (10, 50, 100, and 200 cells per well). Bromelain inhibits AGS, PC3, and MCF7 cells proliferation in such a dose-dependent manner. Determined IC50 to AGS, PC3, and MCF7 cells were 65, 60 and 65μg/ml respectively. At IC50, bromelain significantly suppressed the AGS, PC3, and MCF7 cells colony formation at four treated densities (10, 50, 100 and 200 cells per well). Plating efficiency percentage and cell surviving fraction were decreased after bromelain treatment to AGS, PC3, and MCF7 human cancer cells as a function of initial cell density. The 50, 50 or 100, and 10 or 50 cells per well were considered to be optimum number of initial cell density for AGS, PC3, and MCF7 cells. Cell proliferative and colony formation inhibition are two pathways to in vitro bromelain anticancer effects. The current study displayed a dose-dependent inhibitory effect of bromelain, as well impeding colony formation AGS, PC3, and MCF7 human cancer cells.

The aim of the present study was to construct the eukaryotic expression vector pcDNA3.1/15-PGDH. The vector was used to transfect mouse murine forestomach carcinoma (MFC) cancer cells and observe the effects of 15-hydroxyprostaglandin dehydrogenase (15-PGDH) on the proliferation of MFC. pcDNA3.1/15-PGDH was constructed using gene recombination technology and the vector was used to transfect MFC cells to build a stable transfected cell strain. The expression levels of 15-PGDH in the transfected cells were detected using reverse transcription polymerase chain reaction. Optical Density (OD) values were determined using an MTT assay and used to draw cell growth curves. The effects of 15-PGDH on the proliferation of MFC were observed using a clone formation experiment. Following successful transfection by 15-PGDH, the relative expression levels of 15-PGDH in the MFC/15-PGDH cells were significantly higher (1.06±0.08) (P

A protocol for plant regeneration from cell suspension-derived protoplasts of Populus × beijingensis is described. Protoplasts were isolated from cell suspension cultures 6 d after subculture and further cultured in liquid NH4NO3-free Murashige and Skoog (MS) medium supplemented with 0.6 M glucose, 9.05 μM 2,4-dichlorophenoxyacetic acid, and 0.89 μM 6-benzyladenine at a density of 2 × 105 protoplasts per milliliter. The initial plating efficiency and final plating efficiency recorded after 10 and 30 d reached 33.7 and 1.07%, respectively. The proliferated calli transferred to regeneration medium supplemented with 2.22 μM 6-benzyladenine and 0.54 μM α-naphthaleneacetic acid gave the highest rate of shoot formation (44.4%). All protoplast-derived shoots were able to form roots on half-strength MS medium supplemented with 2.46 μM indole-3-butyric acid.

This article addresses the characteristics of metallic skin layer and the performance of hypophosphite based ultrasonic electroless plating baths during the fabrication of nickel/ceramic microfiltration membranes. The process performance was evaluated in terms of overall conversion and metal plating efficiency, and the metal film was characterised in terms of average film pore size, average effective porosity, film thickness and per cent pore densification. Parametric investigations were carried out for a wide range of initial nickel solution concentrations (0·04–0·16 mol L−1) at two different loading ratios (196 and 393 cm2 L−1) for sonication coupled electroless plating. A simple mass transfer model was developed to provide insights with respect to the pertinent mass transfer limitations. Sonication resulted in faster deposition rate, better efficiency and improved surface morphology. In comparison with conventional and agitated electroless plating baths, ultrasound assisted electroless plating ...

Protoplasts isolated from three accessions of cultivated carrot and 5-day-old protoplast-derived aggregates were subjected to selection to identify somaclonal variants with enhanced tolerance to the fungal disease black rot incited by Alternaria radicina. Different concentrations [1, 2, 3.5, 5, 10, 20, 35 and 50 % (v/v)] of a fungal culture filtrate (FCF) from 2-week-old liquid cultures of A. radicina were used. Protoplasts and aggregates were subjected to short-term selection for a period of 10 days. All FCF concentrations added to the cultures on the day of isolation decreased protoplast survival frequency and plating efficiency, while FCF applied 5 days later inhibited cell divisions in 5–50 % concentrations. The responses of protoplasts to the treatment were genotype dependent. Most R0 plants were regenerated in all accessions from cell lines grown with 1 % FCF, while only a few plants were produced from 2 to 3.5 % FCF-treated cultures of ‘Dolanka’ and the breeding line ‘9304B’, respectively. Nineteen-percent of putative stress-tolerant regenerants were tetraploids, while only 5 % tetraploids were observed in the control. The incidence of unique random amplified polymorphic DNA fragments indicating possible chromosomal rearrangements was low and did not differ among regenerants after selection and those derived from the control. Mobilization of miniature inverted repeat transposable elements was not observed. Some R0 individuals regenerated both from FCF-treated and untreated cultures showed lower susceptibility to A. radicina in a laboratory assay in comparison to control plants grown from seed. Regenerants from FCF-treated cultures showed lower frequency of flowering plants and a higher rate of male sterility. Pollen viability of the putative stress-tolerant regenerants varied over a wide range (6–98 %), independently of in vitro selection conditions. Our data suggest that A. radicina FCF may be feasible for the in vitro selection to generate plants with superior phenotypic performance against A. radicina.

The aim of this work was to develop a semi-selective medium for recovery and enumeration of Pichia guilliermondii strain Z1, a reliable biocontrol agent against postharvest pathogens of citrus fruit, and to assess the population dynamic of this antagonistic yeast on orange fruit in relation to incubation temperature and time of incubation. PDA is the basal medium used in this study which allows the antagonistic strain Z1 source of carbon and nutrients. Different chemicals (thiophanate-methyl, thiabendazole, thiram and imazalil) and antibiotics (hygromycin, tetracyclin, ampicillin and chloramphenicol) have been tested individually and based on the plating efficiency up to 90%, 40 combinations have been assayed between antibiotics and fungicides. Ten combinations proved to be highly selective against citrus pathogens and laboratory microflora, but only one consisting of tetracyclin (1 g/L) and thiabendazole (60 mg/L) was retained based on plating efficiency up to 99%, total selectivity against laboratory microflora, epiphytic microflora from washed orange fruit and lower cost. The semi-selective medium TET-TBZ-PDA has been used to assess the impact of temperature and incubation time on the survival of strain Z1. It appears that population density was significantly influenced by both factors and the highest population size was recorded at 25°C followed by 5 and 35°C, respectively. This strain required a time of adaptation before entering the exponential growth phase with a maximum growth observed at 25°C relative to others. The semi-selective medium TET-TBZ-PDA could be an efficient and valuable way to track the population density of this strain on the surface of orange when applied pre-or postharvest. This semi-selective medium may also aid in reaching a population density allowing a better efficiency in relation to environmental conditions. Key words: Antibiotics, biocontrol, citrus, fungicides, quantification, plating efficiency,Pichia guilliermondii strain Z1, semi-selective medium.