scholarly journals Role of cation-chloride cotransporters, Na/K-pump, and channels in cell water/ionic balance regulation under hyperosmolar conditions: in silico and experimental studies of opposite RVI and AVD responses of U937 cells to hyperosmolar media

2021 ◽  
Author(s):  
Valentina E. Yurinskaya ◽  
Alexey A Vereninov
Keyword(s):  
Animal Cell ◽  
Buoyant Density ◽  
Experimental Studies ◽  
Functional Expression ◽  
Optical Methods ◽  
Ionic Balance ◽  
U937 Cells ◽  
Cell Water ◽  
Monovalent Ions ◽  
Cellular Water

The work provides a modern mathematical description of animal cell electrochemical system under a balanced state and during the transition caused by an increase in external osmolarity, considering all the main ionic pathways in the cell membrane: the sodium pump, K+, Na+, Cl- electroconductive channels and cotransporters NC, KC, and NKCC. The description is applied to experimental data obtained on U937 cells cultured in suspension, which allows the required assays to be performed, including determination of cell water content using buoyant density, cell ion content using flame photometry, and optical methods using flow cytometry. The study of these cells can serve as a useful model for understanding the general mechanisms of regulation of cellular water and ionic balance, which cannot be properly analyzed in many important practical cases, such as ischemic disturbance of cellular ionic and water balance, when cells cannot be isolated. An essential part of the results is the developed software supplied with an executable file, which allows researchers with no programming experience to calculate unidirectional fluxes of monovalent ions through separate pathways and ion-electrochemical gradients that move ions through them, which is important for studying the functional expression of channels and transporters. It is shown how the developed approach is used to reveal changes in channels and transporters underlying the RVI and AVD responses to the hyperosmolar medium in the studied living U937 cells.

scholarly journals Cation-chloride cotransporters, Na/K pump, and channels in cell water and ions regulation: in silico and experimental studies of the U937 cells under stopping the pump and during RVD

2021 ◽  
Author(s):  
Alexey A Vereninov ◽  
Valentina Yurinskaya
Keyword(s):  
Plasma Membrane ◽  
Cell Membrane ◽  
Real Time ◽  
Intracellular Signaling ◽  
Ion Homeostasis ◽  
Regulatory Volume Decrease ◽  
Experimental Studies ◽  
U937 Cells ◽  
Ionic Homeostasis ◽  
Monovalent Ions

Cation-coupled chloride cotransporters play a key role in generating the Cl− electrochemical gradient on the cell membrane which is important for regulation of many cellular processes. However, the cooperation of transporters and channels of the plasma membrane in holding the ionic homeostasis of the whole cell remains poorly characterized because of the lack of a suitable tool for its computation. Our software successfully predicted in real-time changes in the ion homeostasis of U937 cells after stopping the Na/K pump, but so far considered the model with only NC cotransporter. Here the model with all main types of cotransporters is used in computation of the rearrangements of ionic homeostasis due to stopping the pump and associated with the regulatory volume decrease (RVD) of cells swollen in hypoosmolar medium. The parameters obtained for the real U937 cells are used. Successful prediction of changes in ion homeostasis in real-time after stopping the pump using the model with all major cotransporters indicates that the model is reliable. Using this model for analysis RVD showed that there is a "physical" RVD, associated with the time-dependent changes in electrochemical ion gradients, but not with alteration of channels and transporters of the plasma membrane that should be considered in studies of truly active regulatory processes mediated by the intracellular signaling network. The developed software can be useful for calculation of the balance of the partial unidirectional fluxes of monovalent ions across the cell membrane of various cells under various conditions.

Na+/NH4+, Na+/H+ Exchanges and NH3 Movement Across the Gill of Carassius Auratus

1973 ◽  
Vol 58 (1) ◽  
pp. 255-275 ◽  
Author(s):  
J. MAETZ
Keyword(s):  
Carassius Auratus ◽  
External Medium ◽  
Ammonia Excretion ◽  
Alkaline Media ◽  
Ionic Balance ◽  
Free Base ◽  
Acid Media ◽  
Base Form ◽  
Monovalent Ions ◽  
The Law

1. Sodium exchange, ammonia excretion and H+ ion movement across the gill were measured in sodium-depleted Carassius auratus. Sodium uptake and ammonia excretion are considerably enhanced by sodium depletion. In this respect sodium-depleted fish behave as ammonia-loaded fish. 2. A decrease of the external pH by one unit results in a 50-60% decrease in both influx and efflux of sodium. A transient augmentation of the rate of ammonia excretion is observed upon acidification, suggesting an increased permeability of the gill to the free-base form of ammonia. 3. The ionic balance sheet of the movements of the monovalent ions across the gill was drawn up when a permeant co-ion for sodium was absent from the external medium. In both acid and alkaline media the ionic balance was achieved in accordance with the law of electroneutrality of external and internal solutions. 4. Net Na+ uptake was only found to be correlated to NH4+ excretion if H+ ion movements across the gill were taken into account. 5. Ammonia-gradient experiments were performed to test the ability of the gill to excrete ammonia against a gradient of ammonia partial pressure. Ammonia excretion is independent of this gradient in alkaline media when sodium ions are present. In these conditions ammonia moves in the ionic form. In acid media, with Na present in the external medium, ammonia movement is correlated with the pNH3 gradient but uphill movement still occurs. Ammonia moves in both the free-base and the ionized forms. In the absence of external sodium, movements of ammonia obey the law for passive transfer and ammonia moves exclusively in the free-base form, even though the external medium is alkaline.

Investigation of the Flow Structures in Supersonic Free and Impinging Jet Flows

2013 ◽  
Vol 135 (3) ◽  
Author(s):  
C. Chin ◽  
M. Li ◽  
C. Harkin ◽  
T. Rochwerger ◽  
L. Chan ◽  
...  
Keyword(s):  
High Speed ◽  
Numerical Study ◽  
Experimental Studies ◽  
Turbulence Models ◽  
Impinging Jet ◽  
Optical Methods ◽  
Navier Stokes ◽  
Jet Flows ◽  
Shock Cell ◽  
Rans Turbulence Models

A numerical study of compressible jet flows is carried out using Reynolds averaged Navier–Stokes (RANS) turbulence models such as k-ɛ and k-ω-SST. An experimental investigation is performed concurrently using high-speed optical methods such as Schlieren photography and shadowgraphy. Numerical and experimental studies are carried out for the compressible impinging at various impinging angles and nozzle-to-wall distances. The results from both investigations converge remarkably well and agree with experimental data from the open literature. From the flow visualizations of the velocity fields, the RANS simulations accurately model the shock structures within the core jet region. The first shock cell is found to be constraint due to the interaction with the bow-shock structure for nozzle-to-wall distance less than 1.5 nozzle diameter. The results from the current study show that the RANS models utilized are suitable to simulate compressible free jets and impinging jet flows with varying impinging angles.

scholarly journals Attempt to Isolate Elephant Endotheliotropic Herpesvirus (EEHV) Using a Continuous Cell Culture System

Animals ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 2328
Author(s):  
Kornravee Photichai ◽  
Thunyamas Guntawang ◽  
Tidaratt Sittisak ◽  
Varankpicha Kochagul ◽  
Phongsakorn Chuammitri ◽  
...  
Keyword(s):  
Cell Lines ◽  
Dna Polymerase ◽  
Animal Cell ◽  
Hemorrhagic Disease ◽  
U937 Cells ◽  
Tissue Samples ◽  
Continuous Cell Lines ◽  
Cytopathic Effects ◽  
Hct 116

Elephant endotheliotropic herpesvirus (EEHV) infection is known to cause acute fatal hemorrhagic disease, which has killed many young Asian elephants (Elephas maximus). Until recently, in vitro isolation and propagation of the virus have not been successful. This study aimed to isolate and propagate EEHV using continuous cell lines derived from human and/or animal origins. Human cell lines, including EA. hy926, A549, U937, RKO, SW620, HCT-116 and HT-29, and animal cell lines, including CT26.CL25 and sp2/0-Ag14, were investigated in this study. Mixed frozen tissue samples of the heart, lung, liver, spleen and kidney obtained from fatal EEHV1A- or EEHV4-infected cases were homogenized and used for cell inoculation. At 6, 24, 48 and 72 h post infection (hpi), EEHV-inoculated cells were observed for cytopathic effects (CPEs) or were assessed for EEHV infection by immunoperoxidase monolayer assay (IPMA) or quantitative PCR. The results were then compared to those of the mock-infected controls. Replication of EEHV in the tested cells was further determined by immunohistochemistry of cell pellets using anti-EEHV DNA polymerase antibodies or re-inoculated cells with supernatants obtained from passages 2 or 3 of the culture medium. The results reveal that no CPEs were observed in the tested cells, while immunolabeling for EEHV gB was observed in only U937 human myeloid leukemia cells. However, quantitation values of the EEHV terminase gene, as well as those of the EEHV gB or EEHV DNA polymerase proteins in U937 cells, gradually declined from passage 1 to passage 3. The findings of this study indicate that despite poor adaptation in U937 cells, this cell line displays promise and potential to be used for the isolation of EEHV1 and EEHV4 in vitro.

scholarly journals Biomedical applications of Raman and infrared spectroscopy to diagnose tissues

2006 ◽  
Vol 20 (5-6) ◽  
pp. 195-218 ◽  
Author(s):  
C. Krafft ◽  
V. Sergo
Keyword(s):  
Biomedical Applications ◽  
Experimental Studies ◽  
Optical Methods ◽  
Ocular Tissue ◽  
Cervix Uteri ◽  
Mineralized Tissue ◽  
Technical Aspects ◽  
The Past ◽  
Fiber Optical ◽  
Raman And Infrared Spectroscopy

The objective of the article is to review biomedical applications which became possible after the development of sensitive and high throughput Raman and Fourier transform infrared spectrometers in the past decade. Technical aspects of the instrumentation are briefly described. Then the broad range of vibrational spectroscopic applications with the focus on imaging and fiber-optical methods are discussed to study mineralized tissue (bone, teeth), skin, brain, the gastrointestinal tract (mouth, pharynx, esophagus, colon), breast, arteries, cartilage, cervix uteri, the urinary tract (prostate, bladder), lung, ocular tissue, liver, heart and spleen. Experimental studies are summarized demonstrating the possibilities and prospects of these methods in various fields of biodiagnostics to detect and characterize diseases, tumors and other pathologies.

scholarly journals Molecular cloning, sequence analysis, and functional expression of a novel growth regulator, oncostatin M.

1989 ◽  
Vol 9 (7) ◽  
pp. 2847-2853 ◽  
Author(s):  
N Malik ◽  
J C Kallestad ◽  
N L Gunderson ◽  
S D Austin ◽  
M G Neubauer ◽  
...  
Keyword(s):  
Amino Acid ◽  
Sequence Analysis ◽  
Amino Acid Sequence ◽  
Growth Regulator ◽  
Functional Expression ◽  
Lymphoid Cells ◽  
Oncostatin M ◽  
Cdna Clones ◽  
U937 Cells ◽  
Structural And Functional Properties

Oncostatin M is a polypeptide of Mr approximately 28,000 that acts as a growth regulator for many cultured mammalian cells. We report the cDNA and genomic cloning, sequence analysis, and functional expression in heterologous cells of oncostatin M. cDNA clones were isolated from mRNA of U937 cells that had been induced to differentiate into macrophagelike cells by treatment with phorbol 12-myristate 13-acetate, and a genomic clone was also isolated from human brain DNA. Sequence analysis of these clones established the 1,814-base-pair cDNA sequence as well as exon boundaries. This sequence predicted that oncostatin M is synthesized as a 252-amino-acid polypeptide, with a 25-residue hydrophobic sequence resembling a signal peptide at the N terminus. The predicted oncostatin M amino acid sequence shared no homology with other known proteins, but the sequence of the 3' noncoding region of the cDNA contained an A + T-rich stretch with sequence motifs found in the 3' untranslated regions of many cytokine and lymphokine cDNAs. Oncostatin M mRNA of approximately 2 kilobase pairs was detected in phorbol 12-myristate 13-acetate-treated U937 cells and in activated human T cells. Transfection of cDNA encoding the oncostatin M precursor into COS cells resulted in the secretion of proteins with the structural and functional properties of oncostatin M. The unique amino acid sequence, expression by lymphoid cells, and growth-regulatory activities of oncostatin M suggest that it is a novel cytokine.

scholarly journals A Novel Mite-Transmitted Virus with a Divided RNA Genome Closely Associated with Pigeonpea Sterility Mosaic Disease

2003 ◽  
Vol 93 (1) ◽  
pp. 71-81 ◽  
Author(s):  
P. Lava Kumar ◽  
A. Teifion Jones ◽  
D. V. R. Reddy
Keyword(s):  
Polyclonal Antibodies ◽  
Indian Subcontinent ◽  
Buoyant Density ◽  
Experimental Studies ◽  
Cesium Chloride ◽  
Plant Viruses ◽  
Mosaic Disease ◽  
High Plains ◽  
Virus Species ◽  
Eriophyid Mite

The agent of sterility mosaic, a disease that is a major constraint on pigeonpea (Cajanus cajan) production in the Indian subcontinent, is transmitted by the eriophyid mite, Aceria cajani. This agent has remained elusive for decades despite intensive efforts but we report the isolation of highly flexuous filamentous virus-like particles (VLPs) of 3 to 10 nm in width and of undefined lengths from sterility mosaic disease (SMD)-affected pigeonpea plants. Purified VLP preparations from virus-infected pigeonpea and Nicotiana benthamiana had a buoyant density in cesium chloride of 1.22 to 1.23 g cm-3 and contained a major virus-specific protein species of ≈32 kDa and 5 to 7 RNA species of ≈6.8 to 1.1 kb. The sequence of some complementary DNA clones to RNA from purified VLP preparations had no significant matches in database searches. Two oligonucleotide primers derived from one such sequence, when used in reverse transcriptase-polymerase chain reaction assays, amplified a product of 321 bp specifically from SMD-affected pigeonpea plants. Purified VLP preparations were used to produce polyclonal antibodies that, in infected plants, detected the virus using enzyme-linked immuno-sorbent assay (ELISA) and the virus-specific 32-kDa protein in western immunoblotting (WIB). In such assays, the virus was detected consistently in all SMD-affected pigeonpea plant samples from several different locations in India, but not in samples from symptom-free pigeonpea plants from the same locations. In experimental studies, all pigeonpea plants inoculated with viruliferous A. cajani and those plants graft-inoculated with SMD-affected tissue were infected with the virus as assessed by ELISA and WIB, but not any uninfected pigeonpea plants. This virus, tentatively named Pigeonpea sterility mosaic virus (PPSMV), has some properties similar to virus species in the genera Tospovirus and Tenuivirus and with the eriophyid mite-transmitted High plains virus (HPV) but is distinct from these and from all other characterized viruses. The combination of novel properties shown by PPSMV and HPV suggest that they may constitute species in a new genus of plant viruses.

Partial Discharges Registration in Transformer Oil at the ‘Point-Plane’ Electrode System

2014 ◽  
Vol 698 ◽  
pp. 615-620 ◽  
Author(s):  
Aleksandr Bychkov ◽  
Sergey Korobeynikov ◽  
Aleksandr Ovsyannikov
Keyword(s):  
Partial Discharge ◽  
Experimental Studies ◽  
Electrode System ◽  
Partial Discharges ◽  
Transformer Oil ◽  
Optical Methods ◽  
Time Duration ◽  
Negative Point ◽  
Different Types ◽  
System Point

The results of the experimental studies of partial discharges in GK transformer oil with electrode system “point-plane” that provide sharp inhomogeneous electric field are presented. It was found out that two types of partial discharge pulses which differ in form and time duration appeared at the negative point polarity of the needle. The study of different types of pulses was conducted by electric and electro-optical methods of partial discharge registration.

scholarly journals Experimentally determining the effects of water droplets collision when mixing aerosol with gas flow at different heating temperatures

2020 ◽  
Vol 24 (3 Part B) ◽  
pp. 2243-2253
Author(s):  
Olga Vysokomornaya ◽  
Nikita Shlegel ◽  
Pavel Strizhak
Keyword(s):  
Surface Area ◽  
High Speed ◽  
Gas Flow ◽  
Experimental Studies ◽  
Combustion Products ◽  
Optical Methods ◽  
Internal Diameter ◽  
Water Droplets ◽  
Counter Flow ◽  
Droplet Collisions

The article presents the results of experimental studies of the collisions characteristics for water droplets in an aerosol at its entry into the air counter flow. The temperature of the latter ranged from 20?C to 500?C. Experiments were also carried out with the flow of combustion products having a temperature of 800- 850?C. The initial dimensions (radii) of the droplets in the aerosol were 50-1000 ?m. Visualization of the droplet motion in the counter flow of air and combustion products required the use of a hollow cylinder made of quartz glass with a height of 1 m and an internal diameter of 0.15 m, a cross-correlation complex and optical methods (particle image velocimetry, particle tracking velocimetry, interferometric particle imaging). The characteristics of the droplet interaction (size, velocity, total surface area of the liquid before and after) were controlled using a high-speed video camera and tracking algorithms in the TEMA AUTOMOTIVE software package. The main modes of drops interaction have been identified: bounce, coagulation, scatter, and breakup. The statistical information database has been obtained to describe the interaction modes using diagrams, taking into account the ratio of the sizes of colliding drops, velocities of their motion, and an angle between trajectories of motion. The influence of gas temperature on the probabilistic criteria of droplet collisions, as well as the integral criterion characterizing the change in the liquid surface area due to the intensification of droplet collisions in the gas medium has been established.

Sign in / Sign up

Export Citation Format

Share Document