Stationary Cell Research Articles

Demographic Approaches to the Study of Aging on Cell Cultures

Aging organisms die out in accordance with the “Gompertz law,” i.e., the probability of their death increases with age. Survival curve construction is the main tool for gerontologists to study aging and test antiaging drugs. The analysis of survival curves includes obtaining some indices characterizing aging of the population, for example, the average and maximum lifespan, the mortality rate, and the aging rate. Testing geroprotectors can be correctly performed only by obtaining such curves. The dying out of stationary cell populations—bacteria, yeast, and mammalian cell cultures—also occurs in accordance with the Gompertz equation. In this regard, it is reasonable to use the construction of survival curves and their analysis to study the “aging” of non-subcultured cell cultures and testing anti-aging drugs on them. We used this approach in our experiments, due to which we were able to detect the positive anti-aging effect of the Quinton Marine Plasma on stationary phase aging culture of Chinese hamster cells.

U.S. Manufacturers among Leading Suppliers in Growing Stationary Fuel Cell Market

Global demand for stationary fuel cells (a low emission, power generation technology)1 is increasing, with shipments almost doubling during 2012–18. The United States was the largest manufacturer of stationary fuel cells in 2018, and a significant exporter. U.S. manufacturers are likely to benefit from continued global market growth, though there is significant foreign competition and many firms are not yet profitable.

Replicate separation from dead dwarf cells of Stentor coeruleus

IntroductionThe present studies initially show the induction of dwarf forms from the disrupted cells of the large unicellular organism, Stentor coeruleus. The dwarf cells placed in a toxic solution showed evidence of cell death. Within minutes a morphological replicate of the cell separates and subsequently fades. MethodsDehydration of the commercially available Stentor media in deep well slides (n = 9) caused disruption of the large cells. Rehydration with sterile media allowed formation of mobile dwarf forms. The latter (n = 9) placed in a toxic solution lost mobility and showed evidence of cell death, i.e., apoptosis. Deep well slides (n = 9) containing sterile Stentor media were used as controls. ResultsIn the slides following dehydration/ rehydration of the living Stentor media, 7of 9 showed mobile dwarf cells compared to 0 of 9 with the sterile media alone, p < 0.05). Within 8–12 min, the stationary dwarf cell progressively released a morphological replicate of the dead cell which contained entrapped bacteria. Subsequent fading of the replicate allowed dispersion of the bacteria. ConclusionThese findings provide evidence that cell death indicated by apopotosis (blebbing) is followed by a sequence consisting of the progressive separation of a replicate image which is initially visible then becoming a progressively non-visible, faded image.

Evolutionary engineering of Escherichia coli for improved anaerobic growth in minimal medium accelerated lactate production

Anaerobic fermentation is a favorable process for microbial production of bulk chemicals like ethanol and organic acids. Low productivity is the bottleneck of several anaerobic processes which has significant impact on the technique competitiveness of production strain. Improving growth rate of production strain can speed up the total production cycle and may finally increase productivity of anaerobic processes. In this work, evolutionary engineering of wild-type strain Escherichia coli W3110 was adopted to improve anaerobic growth in mineral medium. Significant increases in exponential growth rate and stationary cell density were achieved in evolved strain WE269, and a 96.5% increase in lactate productivity has also been observed in batch fermentation of this strain with M9 minimal medium. Then, an engineered strain for lactate production (BW100) was constructed by using WE269 as a platform and 98.3g/L lactate (with an optical purity of D-lactate above 95%) was produced in a 5-L bioreactor after 48h with a productivity of 2.05g/(L·h). Finally, preliminary investigation demonstrated that mutation in sucD (sucD M245I) (encoding succinyl-CoA synthetase); ilvG (ilvG Δ1bp) (encoding acetolactate synthase 2 catalytic subunit), and rpoB (rpoB T1037P) (encoding RNA polymerase β subunit) significantly improved anaerobic growth of E. coli. Double-gene mutation in ilvG and sucD resumed most of the growth potential of evolved strain WE269. This work suggested that improving anaerobic growth of production host can increase productivity of organic acids like lactate, and specific mutation-enabled improved growth may also be applied to metabolic engineering for production of other bulk chemicals.

Physiological Implications of Microglia-Synapse Interactions.

Microglia are the sole immune responding cells in the central nervous system. Their role as neuroimmune cells in the pathogenesis of various neurodegenerative and infectious diseases of the brain have been extensively studied. Upon brain disease and infection, they adopt an activated phenotype associated with the release of cytokines and neurotrophic factors and resulting in neuroprotective or neurotoxic outcomes. However, microglia are resident also in the healthy or physiological brain, but much less is known about their role(s) in the healthy brain, partly due to technical limitations regarding investigation of these highly reactive cells in the intact brain. Recent developments in molecular probes and in vivo optical imaging techniques has now helped to characterize microglia in the physiological or healthy brain. In vivo two-photon imaging of fluorescently labeled microglia have revealed that they are highly motile cells in the healthy brain, extending and retracting their processes that extend from a largely stationary cell soma. In this chapter, we briefly summarize some of the physiological functions of microglia in the uninjured brain, with a focus on interactions they have with synapses.

건물용 연료전지-보일러 복합설치 안전성능 평가에 관한 연구

건물용 연료전지-보일러 복합설치 안전성능 평가에 관한 연구

Relationship between electrokinetic parameters of red blood cells and acute coronary syndrome

Aim. To assess the state of electrokinetic properties of red blood cells in different variants of the course of acute coronary syndrome (ACS).&#x0D; Materials and methods. The study deals with the assessment of clinical and laboratory-instrumental data from 201 patients with ACS, the results of electrokinetic parameters of red blood cells, their predictors.&#x0D; Results. According to electrokinetic indices of erythrocytes, no reliable differences between the groups were found out. The mean red blood cell oscillation amplitude (MRBCOA) in group 1 was 1.83 ± 0.05, in group 2 – 1.75 ± 0.06 (t = 0.955, p = 0.341). Stationary red blood cell percentage (SRBCP) in patients with NSTE-ACS was 15.6 ± 1.45, in patients with STE-ACS – 13.54 ± 1.64 (t = 0.957, p = 0.339). The methods of multiple disperse and regressive analysis were used. Predictors of SRBCP indices in NSTE-ACS group appeared to be intima-media complex thickness (IMCT), left ventricle diastolic dysfunction, coronary insufficiency index (CII); in STE-ACS group – IMCT, MRBCOA, troponin I and AST levels. In STE-ACS group, predictors of MRBCOA were SRBCP and troponin I level, in NSTE-ACS – no predictors, influencing MRBCOAwere revealed.&#x0D; Conclusions. In NSTE-ACS group, the following predictors, influencing SRBCP were detected:IMCT, CII, LV diastolic dysfunction. In patients of STE-ACS group, there were found the predictors, influencing electrokinetic properties of red blood cells: IMCT, AST, troponin I. In acute coronary syndrome, changes in electrokinetic parameters of red blood cells are mainly influenced by atherosclerotic process, however, in NSTE-ACS, structural-functional status of myocardium is of primary significance, and in STE-ACS – the degree of manifestation of resorptive-necrotic syndrome.

Regenerative processes for Poisson zero polytopes

Abstract Let (Mt:t&gt;0) be a Markov process of tessellations of ℝℓ, and let (𝒞t:t&gt;0) be the process of their zero cells (zero polytopes), which has the same distribution as the corresponding process for Poisson hyperplane tessellations. In the present paper we describe the stationary zero cell process (at𝒞at:t∈ℝ),a&gt;1, in terms of some regenerative structure and we show that it is a Bernoulli flow. An important application is to STIT tessellation processes.

Bacterial biophotons as non-local information carriers: Species-specific spectral characteristics of a stress response.

Studies by Alexander Gurwitsch in the 1920' s with onion root cells revealed the phenomenon of mitogenetic radiation. Subsequent works by Popp, Van Wijk, Quickenden, Tillbury, and Trushin have demonstrated a link between Gurwitsch's mitogenetic radiation and the biophoton, emissions of light correlated with biological processes. The present study seeks to expand upon these and other works to explore whether biophoton emissions of bacterial cultures is used as an information carrier of environmental stress. Bacterial cultures (Escherichia coli and Serratia marcescens) were incubated for 24 hr in 5 ml of nutrient broth to stationary phase and cell densities of ~107 cells/mL. Cultures of E. coli were placed upon a photomultiplier tube housed within a dark box. A second bacterial culture, either E. coli or S. marcescens, was placed in an identical dark box at a distance of 5 m and received injections of hydrogen peroxide. Spectral analyses revealed significant differences in peak frequencies of 7.2, 10.1, and 24.9 Hz in the amplitude modulation of the emitted biophoton signal with respect to whether a peroxide injection occurred or not, and whether the species receiving the injection was E. coli or S. marcescens. These and the subsequent results of discriminant functions suggest that bacteria may release biophotons as a non‐local communication system in response to stress, and that these biophotons are species specific.

Nanostructured Carbon Electrodes for Increased Power Density in Flow Thermo-Electrochemical Generator Heat Sinks

Heat is a by-product of all energy conversion mechanisms. Efforts to utilize and dissipate heat remain a challenge for further development and optimization of energy conversion devices. Stationary thermo-electrochemical cell is a low cost method to harvest heat; however, it suffers from low power density. Flow thermo-electrochemical cell (fTEC) heat sink presents itself as a unique solution as it can simultaneously scavenge and remove heat to maintain devices in the operating range. In this work, multiwalled nanotube (MWNT) electrodes have been used and electrode configuration has been changed to maximize the temperature difference over a small interelectrode separation. As a result, power per unit area of fTEC heat sink has been improved by more than seven-fold to 0.36 W/m2.

Study of Cell Configuration on the Zinc-Air Battery Performance

Zinc air battery has high energy density and low material cost. It has great potential for large-scale energy storage or electrical vehicle applications. Power density is one of the critical factor for these applications. It is heavily depending on the cell configuration. Four different zinc air batteries were design, fabricated, and tested. Cells under investigation are: 1. ORR (oxygen reduction reaction) half-cell, 2. planar cell (parallel electrode separated by a porous separator filled with electrolyte), 3. stationary electrolyte cell (parallel electrode separated by a stationary electrolyte), 4. circulated electrolyte cell (parallel electrode separated by a circulated electrolyte), and 5. stack with 10 cells in series (Parallel electrode separated by stationary electrolyte and ten cells are laminated stacking together). Their maximum power densities were varied from mW cm-2 to hundred mW cm-2. We found that the thickness of cathode, discharge of gas bubble in the electrolyte, internal resistance, and contact resistance, etc. are playing an important factor on the cell power density. Figure 1

PaPro1 and IDC4, Two Genes Controlling Stationary Phase, Sexual Development and Cell Degeneration in Podospora anserina.

Filamentous fungi frequently undergo bistable phenotypic switches. Crippled Growth of Podospora anserina is one such bistable switch, which seems to rely upon the mis-activation of a self-regulated PaMpk1 MAP kinase regulatory pathway. Here, we identify two new partners of this pathway: PaPro1, a transcription factor orthologous to Sordaria macrospora pro1 and Neurospora crassa ADV-1, and IDC4, a protein with an AIM24 domain. Both PaPro1 and IDC4 regulate stationary phase features, as described for the other actors of the PaMpk1 signaling pathway. However, PaPro1 is also involved in the control of fertilization by activating the transcription of the HMG8 and the mating type transcription factors, as well as the sexual pheromones and receptor genes. The roles of two components of the STRIPAK complex were also investigated by inactivating their encoding genes: PaPro22 and PaPro45. The mutants of these genes were found to have the same phenotypes as PaPro1 and IDC4 mutants as well as additional phenotypes including slow growth, abnormally shaped hyphae, pigment accumulation and blockage of the zygotic tissue development, indicating that the STRIPAK complex regulates, in addition to the PaMpk1 one, other pathways in P. anserina. Overall, the mutants of these four genes confirm the model by which Crippled Growth is due to the abnormal activation of the PaMpk1 MAP kinase cascade.

Abstract 3156: C1orf116, an uncharacterized gene with unknown function, restricts movement capacity of would-be metastatic cells through modulation of tight junctions

Abstract Lethal metastases are the end-result of a cancer cell that escapes the primary tumor and eventually invades a secondary site. Thus, in order for a cancer cell to metastasize from the primary tumor, it must move. In a critical early event of metastasis, a stationary epithelial cell undergoes a dramatic transmogrification event to gain mesenchymal-like features, most notably, ability to move. The full repertoire of genes regulating this switching event and inducing potential metastatic cell movement in the primary tumor remains incomplete. Using a multi-study discovery analysis (doi: 10.1186/s12885-017-3413-3), we identified C1orf116 as a novel candidate driver of an epithelial phenotype. C1orf116 is a highly conserved unnamed gene of unknown function. It is highly expressed in proliferative tissues (e.g. skin, lung, esophagus; GTeX) and is associated with an epithelial gene expression signature in cancer cell lines (CCLE). To determine if C1orf116 is a driver of epithelial phenotype, we overexpressed C1orf116 in C1orf116low PC3 cells (PC3-oe). PC3-oe demonstrated distinctive cobblestone epithelial cellular morphology and increased epithelial marker expression (e.g. OVOL1, CDH1, ESRP). Knockdown of C1orf116 expression in C1orf116high C42b cells (C42b-kd) resulted in decreased expression of classical epithelial markers. This data indicates that C1orf116 was sufficient to induce an epithelial transition. The essential characteristic of a lethal emigrant cell that may give rise to a clinical metastasis is gaining the capacity to move. We found that PC3-oe cultures had significantly reduced motility compared to control, suggesting that C1orf116 restricts motility. Cellular motility is influenced by many factors, including cell-cell adhesion. We observed that PC-oe established cell-cell contacts more rapidly than control cells and hypothesized that C1orf116 promoted cell-cell adhesion. PC3-oe cells had dramatically increased expression of tight junction complex proteins (e.g. ZO-1, ZO2, CLDN1). In contrast, C42b-kd cells had reduced expression of these tight junction components. These data demonstrate that C1orf116 promotes tight junctions and inhibits cell movement, thereby restricting the active emigrant phenotype of potentially metastatic cells. The putative metastasis suppressor role of C1orf116 is further substantiated by clinical data. C1orf116 expression is reduced in radical prostatectomy specimens from patients who progress with a biochemical recurrence compared to those who do not recur. Similarly, C1orf116 expression is decreased in prostate cancer metastatic lesions compared to primary tumor. This work shows the critical role of the overlooked gene and protein C1orf116. Further knowledge of how movement is restricted in the primary tumor provides valuable insight into the lethal metastatic process. Citation Format: Sarah R. Amend, Kenneth J. Pienta. C1orf116, an uncharacterized gene with unknown function, restricts movement capacity of would-be metastatic cells through modulation of tight junctions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3156.

A Coxiella burnetii phospholipase A homolog pldA is required for optimal growth in macrophages and developmental form lipid remodeling

BackgroundMany gram-negative bacteria produce an outer membrane phospholipase A (PldA) that plays an important role in outer membrane function and is associated with virulence.ResultsIn the current study, we characterized a pldA mutant of Coxiella burnetii, an intracellular gram-negative pathogen and the agent of human Q fever. The C. burnetti pldA open reading frame directs synthesis of a protein with conserved PldA active site residues. A C. burnetii ΔpldA deletion mutant had a significant growth defect in THP-1 macrophages, but not axenic medium, that was rescued by complementation. Thin layer chromatography was employed to assess whether pldA plays a role in remodeling membrane lipids during C. burnetii morphological differentiation. Extracted lipids were analyzed from replicating, logarithmic phase large cell variants (LCVs), non-replicating, stationary phase small cell variants (SCVs), and a mixture of LCVs and SCVs. Similar to Escherichia coli, all three forms contained cardiolipin (CL), phosphatidylglycerol (PG) and phosphatidylethanolamine (PE). However, PE and PG were present in lower quantities in the SCV while three additional lipid species were present in higher quantities. Co-migration with standards tentatively identified two of the three SCV-enriched lipids as lyso-phosphatidylethanolamine, a breakdown product of PE, and free fatty acids, which are generally toxic to bacteria. Developmental form lipid modifications required the activity of PldA.ConclusionsCollectively, these results indicate developmentally-regulated lipid synthesis by C. burnetii contributes to colonization of macrophages and may contribute to the environmental stability and the distinct biological properties of the SCV.

CONCENTRATION AND PURIFICATION OF THE PECTIN EXTRACTS BY ULTRAFILTRATION METHOD

The work is devoted to the study of pectin extracts ultrafiltration using different types of membranes. Pectic extracts are the solutions of high molecular weight polysaccharides (MM from 15 to 50 kDa), which are destroyed in the process of heat treatment in the vacuum evaporating apparature, that leads to a significant deterioration in the quality of pectin. The impurity components in the pectin extract of a low molecular weight, which makes it possible to use ultrafiltration for both concentration and purification from ballast substances, that is very important. The objects of research were pectin extracts obtained from citrus raw materials; the membranes of the type UAM-100 and 200A were used as filter membranes. The results of the study have been shown, that the use of UAM-100 membranes in a stationary cell allows to remove up to 20% of the solution within 20 minutes, and with - stirring up to 27%. The use of membranes with large openings (up to 200 A) makes it possible to activate the process of the separation and concentration of pectin extracts: the degree of concentration reaches up 45%, the content of solids in the concentrate increases up to 7.6%, and the content of pectin to up 6.4%, that is in 1,7 times. However, the loss of low molecular weight pectin in the filtrate is about 4%. A comparative analysis of the results of research has shown a high productivity with the use of the capron membrane 200A, as well as on the qualitative parameters of the pectins, released from the concentrates, the higher parameters for the uroconid component, molecular weight, pectin jelly strength and complexing ability were noted for pectins isolated using the UAM- 100 membrane.

Application of toluene LIF to transonic nozzle flows to identify zones of incomplete molecular mixing.

Toluene laser-induced fluorescence (LIF) has been applied to image the mixing deficit on the molecular level in the transonic wake of two different blunt-body injectors in a compressible accelerated nozzle flow. A single-color excitation and two-color detection scheme is employed to measure the signal red-shift caused by the quenching effect of molecular oxygen on the fluorescence of toluene, which reduces and red-shifts the LIF signal if both substances interact on a molecular level. To this end, toluene is injected alternatingly with O2-contaning and O2-free carrier gas into the air main flow. Differences of both signals mark regions where mixing on molecular level is incomplete. A zone of molecular mixing deficit extending several millimeters in stream-wise direction is identified. The effect of local variations in temperature on the sensitivity of this technique is discussed using photo-physical data measured in a stationary low-temperature cell.

Synergy of two fatty acids as additives on lubricity of a nematic liquid crystal 5CB

AbstractIn examining the lubricity of a nematic liquid crystal (4‐pentyl‐4′‐cyanobiphenyl [5CB]), synergy of 2 fatty acids (stearic acid [C18] and behenic acid [C22]) as additives was observed. In “friction measurements” using a ball‐on‐plate‐type tribometer, the friction coefficient for the “mixed‐additive” lubricant (5CB‐C18‐C22) was lower than those for the “single‐additive” lubricants (5CB‐C18 and 5CB‐C22). This synergy mechanism was discussed with considering the results obtained in other tests: “orientation observations” in a stationary cell using polarising microscopy and “surface topography measurements” of adsorbed films using atomic force microscopy. As a result, it is believed that the microscopic patterned surface formed by the mixed additive strengthens the surface anchoring of 5CB, which leads to increase in the effective viscosity of 5CB and therefore to decrease in the macroscopic friction coefficient in the mixed lubrication regime.

LCA分析とDEAを用いた家庭用燃料電池システムにおける環境性及び性能を考慮した指標（FC-DEA）の提案

LCA分析とDEAを用いた家庭用燃料電池システムにおける環境性及び性能を考慮した指標（FC-DEA）の提案

INHIBITORY ACTIVITY OF COBALT(II)–MORIN COMPLEX AGAINST THE REPLICATION OF DENGUE VIRUS TYPE 2

Dengue virus (DENV) is a significant pathogen emerging worldwide as a cause of infectious disease. Antidengue treatments are urgently required to control the emergence of dengue. DENV is a mosquito-borne disease responsible for acute systemic diseases and serious health conditions. DENVs were distributed in the tropical and sub-tropical areas and transmitted to humans by Aedes agypty and Aedes albopictus. Dengue vaccine or antiviral has not yet been clinically approved for humans, even though there have been great efforts toward this end. Antiviral activity against DENV is an important alternative for the characterization and development of drugs. Metal–organic compounds were reported to exhibit fungicidal, bactericidal, and antiviral activities its inhibitory activity was not significant, at high concentration it was more toxic to replicating cells than to stationary cell monolayers of Vero cells. The aim of this study is to investigate the antiviral effects of Cobalt(II)–Morin complex. This compound was further investigated for its inhibitory effect on the replication of DENV-2 in Vero cells. The replication of DENV was measured by enzyme-linked immunosorbent assay and the value of selectivity index (SI). SI was determined as the ratio of the 50% cytotoxic concentration (CC50) to the 50% inhibitory concentration (IC50). The IC50 value of the Cobalt(II)–Morin complex for DENV-2 was 3.08 µg/ml, and the CC50 value of the complex for Vero cells was 3.36 µg/ml; thus, the SI value was 1.09. The results of this study demonstrate the antidengue serotype 2 inhibitory activity of Cobalt(II)–Morin complex and its high toxicity in Vero cells. Further studies are not required before Co(II)–Morin can be applied in the treatment of DENV-2 infections.

Enhanced cell-surface display of a heterologous protein using SED1 anchoring system in SED1-disrupted Saccharomyces cerevisiae strain

Yeast displaying enzymes on the cell surface are used for developing whole-cell biocatalysts. High enzyme activity on the cell surface is required in certain applications such as direct ethanol production from lignocellulosic materials. However, the cell surface enzyme activity is limited by several factors, one of which is the protein amount of the yeast cell wall. In this study, we attempted to improve the incorporation capacity of a displayed heterologous enzyme by disrupting a native cell-wall protein. β-Glucosidase (BGL1) from Aspergillus aculeatus was fused with Saccharomyces cerevisiae Sed1 and displayed on the cell surface of S.cerevisiae BY4741 strain and its SED1 disruptant. Sed1 is one of the most abundant stationary phase yeast cell wall protein. A time course analysis revealed that BGL1 activity of the control strain reached saturation after 48h of cultivation. In contrast, the BGL1 activity of the SED1 disruptant increased until 72h of cultivation and was 22% higher than that of the control strain. We also performed relative quantification of cell wall proteins of these strains by nanoscale ultra pressure liquid chromatography electrospray ionization quadrupole time-of-flight tandem mass spectrometry (nano-UPLC-MSE). The amount of the cell wall-associated BGL1 per unit dry cell-weight of the SED1 disruptant was 19% higher than that of the control strain. These results suggested that the incorporation capacity of the cell wall for BGL1 was increased by disruption of SED1. Disruption of SED1 would be a promising approach for improving display efficiency of heterologous protein fused with Sed1.