scholarly journals Elemental Composition and Cell Mass Quantification of Cultured Thraustochytrids Unveil Their Large Contribution to Marine Carbon Pool

Marine Drugs ◽  
2021 ◽  
Vol 19 (9) ◽  
pp. 493
Author(s):  
Biswarup Sen ◽  
Jiaqian Li ◽  
Lyu Lu ◽  
Mohan Bai ◽  
Yaodong He ◽  
...  
Keyword(s):  
Stationary Phase ◽  
Cell Volume ◽  
Cell Mass ◽  
Lower Percentage ◽  
Large Contribution ◽  
Growth Phases ◽  
Current Estimate ◽  
Cell Content ◽  
Marine Habitats ◽  
C And N

The element stoichiometry of bacteria has received considerable attention because of their significant role in marine ecosystems. However, relatively little is known about the composition of major structural elements of the unicellular heterotrophic protists—thraustochytrids, despite their widely recognized contribution to marine nutrient cycling. Here, we analyze the cell volume and elemental C, N, H, and S cell content of seven cultured thraustochytrids, isolated from different marine habitats, in the exponential and stationary growth phases. We further derive the relationships between the cell volume and elemental C and N content of the cultured thraustochytrids. The cell volumes varied significantly (p < 0.001) among the isolates, with median values of 96.9 and 212.5 μm3 in the exponential and stationary phases, respectively. Our results showed a significantly higher percentage of C (64.0 to 67.5) and H (9.9 to 13.2) but a lower percentage of N (1.86 to 2.16) and S (0.34 to 0.91) in the stationary phase, along with marked variations of C and N fractions among isolates in the exponential phase. The cell C (5.7 to 203.7 pg) and N (0.65 to 6.1 pg) content exhibited a significant (p < 0.001) linear relationship with the cell volume (27.7 to 510 μm3). On further analysis of the relationship across the two growth phases, we found the equation (cell C (pg) = 0.356 × cell volume (μm3) + 20.922) for stationary phase cells more appropriate for C estimation of natural thraustochytrids. This study provides the first experimental evidence of higher cell C density than the current estimate and relatively larger C contribution of thraustochytrids than bacteria to the marine organic pool.

287 ACTIVE MITOCHONDRIA ARE PRESENT IN MOUSE AND MONKEY EMBRYONIC STEM CELL LINES

2008 ◽  
Vol 20 (1) ◽  
pp. 223 ◽  
Author(s):  
T. Lonergan ◽  
A. Harvey ◽  
J. Zhao ◽  
B. Bavister ◽  
C. Brenner
Keyword(s):  
Cell Lines ◽  
Complex I ◽  
Inner Cell Mass ◽  
Es Cells ◽  
Cell Mass ◽  
Embryonic Stem ◽  
Mouse Fibroblast ◽  
Es Cell ◽  
Cell Content ◽  
Stem Cell Lines

The inner cell mass (ICM) of the blastocyst develops into the fetus after uterine implantation. Prior to implantation, ICM cells synthesize ATP by glycolytic reactions. We now report that cells of the ICM in 3.5-day-old mouse embryos have too few mitochondria to be visualized with either Mitotracker red (active mitochondria) or an antibody against complex I of OXPHOS. By comparison, all of the surrounding trophectoderm cells reveal numerous mitochondria throughout their cytoplasm. It has largely been assumed that embryonic stem (ES) stem cells derived from the ICM also have few mitochondria, and that replication of mitochondria in the ES cells does not begin until they commence differentiation. We further report that mouse E14 ES cells and monkey ORMES 7 ES cells have considerable numbers of active mitochondria when cultured under standard conditions, i.e., 5% CO2 in air. Both the mouse E14 and monkey ES cell lines expressed two markers of undifferentiated cells, Oct-4 and SSEA-4, and monkey ES cells expressed the undifferentiated cell marker Nanog; however, Oct-4 is nonspecific in monkey ES cells because trophectoderm also expresses this marker, unlike in mice. Ninety-nine percent of the E14 cells examined, and 100% of the ORMES 7 cells, have a visible mitochondrial mass when stained with either Mitoracker red or with an antibody against OXPHOS complex I. The ATP content in the mouse E14 cells (4.13 pmoles ATP/cell) is not significantly different (P = 0.76) from that in a mouse fibroblast control (3.75 pmoles ATP/cell). Cells of the monkey ORMES 7 cell line had 61% of the ATP/cell content (7.55 pmoles ATP/cell) compared to the monkey fibroblast control (12.38 pmoles ATP/cell). Both cell lines expressed two proteins believed to indicate competence of mitochondria to replicate: PolG, the polymerase used to replicate the mitochondrial genome, and TFAM, a nuclear-encoded transcription factor reported to regulate several aspects of mitochondrial function. Both proteins were found to co-localize in the mitochondria. We conclude that when the ICMs are isolated from blastocysts and used to establish these two ES cell lines in cell culture, mitochondrial biosynthesis is activated.

scholarly journals ON THE PLASTIDS, MITOCHONDRIA, AND OTHER CELL CONSTITUENTS DURING OÖGENESIS OF A PLANT

1966 ◽  
Vol 28 (3) ◽  
pp. 527-543 ◽  
Author(s):  
Lothar Diers
Keyword(s):  
Electron Microscopy ◽  
Cell Volume ◽  
Mother Cell ◽  
Developmental Stages ◽  
Three Dimensional ◽  
Egg Cell ◽  
Lipid Bodies ◽  
Growth Phases ◽  
Quantitative Investigation ◽  
Nucleus Volume

In the liverwort Sphaerocarpus donnellii Aust., the behavior of the cell constituents, especially of mitochondria and plastids, was studied by electron microscopy during the development of the egg and its preceding cells. A degeneration and elimination of mitochondria and plastids was not found in any of the developmental stages. In all growth phases of the archegonium, the plastids may deposit starch which becomes especially frequent in the maturing egg cell. No indications have been observed that new mitochondria or plastids generate from the nuclear evaginations, which often penetrate deeply into the cytoplasm of the maturing and fully developed eggs. A quantitative investigation based on general micrographs elucidates the numerical aspects of the cell constituents during oögenesis. With the increase of cell volume, the numbers of dictyosomes, mitochondria, plastids, and lipid bodies increase. From the stages of the mother cell of the axial row up to that of the mature egg, the cell volume enlarges about 8 times and the nucleus volume about 15 times. Simultaneously, the numbers of mitochondria and plastids increase up to 8 to 15 times. On the basis of these findings, mitochondria and plastids with three-dimensional narrow constrictions are interpreted as divisional stages.

scholarly journals The Response of Duck Erythrocytes to Hypertonic Media

1971 ◽  
Vol 58 (4) ◽  
pp. 396-412 ◽  
Author(s):  
Floyd M. Kregenow
Keyword(s):  
Cell Volume ◽  
Cell Size ◽  
Hypertonic Solution ◽  
Electrochemical Gradient ◽  
Cell Content ◽  
Bathing Medium ◽  
Instantaneous Phase ◽  
Controlling Mechanism ◽  
Pump Mechanism ◽  
The One

The addition of a hypertonic bathing medium to duck erythrocytes results in an initial instantaneous phase of osmotic shrinkage and, when the [K]o of the hypertonic solution is larger than "normal," in a second, more prolonged phase, the volume regulatory phase. During the latter, which also requires extracellular Na, the cells swell until they approach their initial isotonic volume. The increase in cell volume during the volume regulatory phase is accomplished by a gain in the cell content of K, Cl, and H2O. There is also a smaller increase in the Na content of the cell. Potassium is accumulated against an electrochemical gradient and is therefore actively transported into the cell. This accumulation is associated with an increase, although dissimilar, in both K influx and efflux. Changes in cell size during the volume regulatory phase are not altered by 10-4 M ouabain, although this concentration of ouabain does change the cellular cation content. The response is independent of any effect of norepinephrine. The changes in cell size during the volume regulatory phase are discussed as the product of a volume controlling mechanism identical in principle to the one reported in the previous paper which controls cell volume in hypotonic media. Similarly, this mechanism can regulate cell size, when the Na-K exchange, ouabain-inhibitable pump mechanism is blocked.

scholarly journals Technical-Scale Production of Cyanophycin with Recombinant Strains of Escherichia coli

2002 ◽  
Vol 68 (7) ◽  
pp. 3377-3384 ◽  
Author(s):  
Kay M. Frey ◽  
Fred B. Oppermann-Sanio ◽  
Holger Schmidt ◽  
Alexander Steinbüchel
Keyword(s):  
Escherichia Coli ◽  
Large Scale ◽  
Cell Mass ◽  
Extraction Procedure ◽  
Temperature Shift ◽  
Exponential Growth Phase ◽  
Acid Extraction ◽  
Scale Production ◽  
Mineral Salts ◽  
Cell Content

ABSTRACT By the use of Escherichia coli DH1 harboring cphA from Synechocystis sp. strain PCC6803, large-scale production of cyanophycin at 30- and 500-liter culture volumes was established. Transcription of cphA was controlled by the thermosensitive cI857 repressor, which enabled induction of cphA by a simple temperature shift in the culture fluid. Maximum cyanophycin cell content of up to 24% (wt/wt) of cellular dry matter was obtained by induction in the early exponential growth phase and cultivation of the cells in terrific broth complex medium. Synthesis of cyanophycin was found to be strongly dependent on the presence of complex components, and in mineral salts medium the cells synthesized and accumulated cyanophycin only if Casamino Acids were added. Cultivations were done at the 500-liter scale, allowing the provision of cell mass for the preparation of cyanophycin at the kilogram scale. Isolation of cyanophycin was achieved by a new acid extraction procedure which allowed large-scale purification of the polyamide from whole cells.

scholarly journals Cellular electrolyte and volume changes induced by acidosis in the rabbit proximal straight tubule.

1991 ◽  
Vol 2 (5) ◽  
pp. 1030-1040
Author(s):  
L P Sullivan ◽  
D P Wallace ◽  
R L Clancy ◽  
C Lechene ◽  
J J Grantham
Keyword(s):  
Cell Volume ◽  
Ph Effect ◽  
Basolateral Membrane ◽  
Control Level ◽  
Cell Swelling ◽  
High Pco2 ◽  
Cell Content ◽  
Cell Ph ◽  
Additional Cell ◽  
Altered Cell

Cellular acidosis induced either by high Pco2 or by low HCO3- concentrations has been shown to cause cell swelling in isolated, lumen-collapsed, S2 segments of the rabbit proximal tubule (Sullivan et al., Am J Physiol 1990; 258: F831-F839). The swelling is not followed by a volume regulatory response. The ionic basis of the swelling has been investigated by measurement of the cellular K+, Na+, and Cl- content (electron probe) and HCO3- concentration (pH-sensitive fluorescent dye). Cell content of K+, Na+, and Cl- was expressed as a ratio to P content. Exposure to 15% CO2 increased K/P from 0.98 to 1.16, Cl/P from 0.14 to 0.20, and Na/P from 0.09 to 0.11. Cell (HCO3-) increased from 22 to 32 mM. Reduction in bath (HCO3-) from 25 to 5 mM reduced cell (HCO3-) from 24 to 8 mM and increased K/P from 0.75 to 0.90. Na/P fell from 0.13 to 0.09, and Cl/P fell from 0.15 to 0.12. Thus, swelling resulting from acidosis induced by high CO2 was accompanied by an accumulation of K+, Cl-, and HCO3-; that resulting from acidosis induced by a fall in (HCO3-) was combined with an accumulation of K+ and an unidentified anion. To determine if the swelling induced by a fall in pH might be coupled with depolarization of the basolateral membrane, the effect of 1 mM barium was tested. Barium caused cell volume to increase 10.2%. Cell pH rose from 7.38 to 7.56, K/P increased from 0.63 to 0.73, Na/P did not change, and Cl/P rose from 0.17 to 0.20. Cell (HCO3-) increased 10.4 mM. When the pH of the barium-treated tissue was reduced to 7.02 by raising Pco2, additional cell swelling and accumulation of K+ occurred. The effect on cell volume of a reduction of bath (HCO3-) from 25 to 5 mM at constant bath pH was determined. Cell pH was not altered. Cell volume decreased 3% initially and then returned to the control level. When the bath (HCO3-) was restored to 25 mM, cell volume increased 3.9% and then returned to the baseline. Thus, volume regulation was not impaired. It was concluded that a fall in cell pH induces swelling, and this is coupled with an accumulation of K+. This is probably the result of a pH effect on barium-sensitive and barium-insensitive K+ conductance pathways. The nature of the anions that balance the gain in K+ depends on the means used to induce acidosis.

scholarly journals Plasmid transfer in planktonically-grown cultures and biofilms

2021 ◽  
Author(s):  
Farhana Shamsad
Keyword(s):  
Cell Growth ◽  
Stationary Phase ◽  
Growth Phase ◽  
Nutrient Concentrations ◽  
Bacterial Cells ◽  
Growth Phases ◽  
Rt Pcr ◽  
Lower Growth ◽  
Donor Cells ◽  
Significant Difference

Conjugation of plasmids is widespread among bacteria and contributes to the spread of antibiotic resistance. In the natural environment, microorganisms predominantly exist in the form of biofilms or other bioaggregates, where they may be exposed to contaminants such as antibiotics, at subinhibitory concentrations. Bacterial cells in older biofilms have lower growth activity due to oxygen and nutrient limitation in the deeper layers of the biofilms. In batch culture, population growth eventually ceases during the stationary phase. Thus, the steady state of biofilms may resemble stationary growth phase cultures. Our objectives were to study (i) the effect of cell growth phases and (ii) subinhibitory and minimum inhibitory concentrations (MIC) of antibiotics on transconjugant formation in both batch cultures and biofilms. Additionally, (iii) the effect of variable nutrient concentrations on MIC was investigated and (iv) an optimization of RT-PCR method for the detection of traA gene (which encodes pilus biosynthesis) expression was carried out. To study the effect of cell growth phases on transconjugant formation, plate matings were carried out utilizing planktonic cultures grown to exponential or stationary phase of donor and recipient strains. The results showed that transconjugant abundance was the highest (20± 0.08%) when both plasmid donor and recipient cells were grown to the stationary phase. However, the growth phase of the donor did not seem to play a role in biofilms. When donor cells were harvested from either the exponential or stationary phase of growth, and inoculated into 24 h old recipient biofilms, there was no statistically significant difference between transconjugant abundance. A higher percentage of transconjugants was detected in plate matings when the donor was exposed to 0.5× minimum inhibitory concentration of gentamicin and additionally challenged with gentamicin at MIC. In biofilms, transconjugant formation was not enhanced when the donor cells were grown with 0.5× MIC gentamicin, and 0.5× MIC gentamicin was added to the biofilms. A decrease in nutrient concentration was associated with a decrease in the MIC. traA expression, detected using RT-PCR in plasmid donor cells grown to early exponential and late exponential phases did not coincide with an increase in transconjugants.

scholarly journals 3D quantification of changes in pancreatic islets in mouse models of diabetes type I and II

2020 ◽  
Vol 13 (12) ◽  
pp. dmm045351
Author(s):  
Urmas Roostalu ◽  
Jacob Lercke Skytte ◽  
Casper Gravesen Salinas ◽  
Thomas Klein ◽  
Niels Vrang ◽  
...  
Keyword(s):  
Blood Glucose ◽  
Beta Cell ◽  
Cell Volume ◽  
Beta Cells ◽  
Mouse Models ◽  
Cell Mass ◽  
Beta Cell Mass ◽  
Light Sheet ◽  
Type I ◽  
Beta Cell Volume

ABSTRACTDiabetes is characterized by rising levels of blood glucose and is often associated with a progressive loss of insulin-producing beta cells. Recent studies have demonstrated that it is possible to regenerate new beta cells through proliferation of existing beta cells or trans-differentiation of other cell types into beta cells, raising hope that diabetes can be cured through restoration of functional beta cell mass. Efficient quantification of beta cell mass and islet characteristics is needed to enhance drug discovery for diabetes. Here, we report a 3D quantitative imaging platform for unbiased evaluation of changes in islets in mouse models of type I and II diabetes. To determine whether the method can detect pharmacologically induced changes in beta cell volume, mice were treated for 14 days with either vehicle or the insulin receptor antagonist S961 (2.4 nmol/day) using osmotic minipumps. Mice treated with S961 displayed increased blood glucose and insulin levels. Light-sheet imaging of insulin and Ki67 (also known as Mki67)-immunostained pancreata revealed a 43% increase in beta cell volume and 21% increase in islet number. S961 treatment resulted in an increase in islets positive for the cell proliferation marker Ki67, suggesting that proliferation of existing beta cells underlies the expansion of total beta cell volume. Using light-sheet imaging of a non-obese diabetic mouse model of type I diabetes, we also characterized the infiltration of CD45 (also known as PTPRC)-labeled leukocytes in islets. At 14 weeks, 40% of the small islets, but more than 80% of large islets, showed leukocyte infiltration. These results demonstrate how quantitative light-sheet imaging can capture changes in individual islets to help pharmacological research in diabetes.

scholarly journals MORPHOLOGICAL AND MORPHOMETRICAL FEATURES IN DUNALIELLA SALINA (CHLAMYDOMONADALES, DUNALIELLACEAE) DURING THE TWO-PHASE CULTIVATION MODE

2019 ◽  
Vol 22 ◽  
pp. 157-165
Author(s):  
Andrei B. Borovkov ◽  
Irina N. Gudvilovich ◽  
Olga A. Memetshaeva ◽  
Anna L. Avsiyan ◽  
Alexander S. Lelekov ◽  
...  
Keyword(s):  
Cell Volume ◽  
Dunaliella Salina ◽  
Linear Growth ◽  
Cell Number ◽  
Two Phase ◽  
Cell Content ◽  
Crimean Peninsula ◽  
Cell Parameters ◽  
Maximum Productivity ◽  
Salina Cell

The paper presents studies of morphological and morphometrical characteristics of green halophilic carotenogenic microalga Dunaliella salina (Dunal) Teodoresco, 1905, from the south-west region of Crimean Peninsula. D. salina was cultivated in two-phase mode under conditions of natural illumination at the premises of A.O. Kovalevsky Institute of Biology of the Southern Seas of RAS (IBSS), Sevastopol, Russia. The maximum D. salina cell density was 1.69·106 cell/ml in the “green” phase and 0.84·106 cell/ml in the “red” growth phase. The maximum productivity by cell number reached 0.15 ∙ 106 cell/(ml·day)in the “green” phase while it was lower by 73% in the “red” phase (0.04∙106 cell/(ml·day). Along with the maximum productivity, linear growth stage in the first phase was characterized by a maximum fraction of small (up to 500 μm3 in volume) cells (about 15-29%) and a decrease in cell volume by 40-45% as compared with initial value. The mean of D. salina cell volume in the “red” phase was 30% higher than in the “green” phase. At the same time, the large cell fraction in the “red” phase was consistently high (15-35%). The patterns of change in morphological and morphometrical cell parameters were in accordance to stage and conditions of growth. Thus, cell elongation was noted in the stage of linear growth, while under unfavorable conditions at growth-declining stage cells became more round-shaped, with orange and tile-red coloration and granulation of cell content. It was shown that morphological and morphometric cell parameters can serve as additional criteria for assessment of physiological condition in D. salina culture. The experiment demonstrated the prospects for two-stage D. salina cultivation in Crimea.

scholarly journals Metabolic preference of nitrate over oxygen as an electron acceptor in foraminifera from the Peruvian oxygen minimum zone

2019 ◽  
Vol 116 (8) ◽  
pp. 2860-2865 ◽  
Author(s):  
Nicolaas Glock ◽  
Alexandra-Sophie Roy ◽  
Dennis Romero ◽  
Tanita Wein ◽  
Julia Weissenbach ◽  
...  
Keyword(s):  
Cell Volume ◽  
Electron Acceptor ◽  
Oxygen Minimum Zone ◽  
Diverse Range ◽  
Mean Cell Volume ◽  
Respiration Rates ◽  
Marine Habitats ◽  
Biogeochemical Models ◽  
Minimum Zone ◽  
Oxygen Minimum

Benthic foraminifera populate a diverse range of marine habitats. Their ability to use alternative electron acceptors—nitrate (NO3−) or oxygen (O2)—makes them important mediators of benthic nitrogen cycling. Nevertheless, the metabolic scaling of the two alternative respiration pathways and the environmental determinants of foraminiferal denitrification rates are yet unknown. We measured denitrification and O2respiration rates for 10 benthic foraminifer species sampled in the Peruvian oxygen minimum zone (OMZ). Denitrification and O2respiration rates significantly scale sublinearly with the cell volume. The scaling is lower for O2respiration than for denitrification, indicating that NO3−metabolism during denitrification is more efficient than O2metabolism during aerobic respiration in foraminifera from the Peruvian OMZ. The negative correlation of the O2respiration rate with the surface/volume ratio is steeper than for the denitrification rate. This is likely explained by the presence of an intracellular NO3−storage in denitrifying foraminifera. Furthermore, we observe an increasing mean cell volume of the Peruvian foraminifera, under higher NO3−availability. This suggests that the cell size of denitrifying foraminifera is not limited by O2but rather by NO3−availability. Based on our findings, we develop a mathematical formulation of foraminiferal cell volume as a predictor of respiration and denitrification rates, which can further constrain foraminiferal biogeochemical cycling in biogeochemical models. Our findings show that NO3−is the preferred electron acceptor in foraminifera from the OMZ, where the foraminiferal contribution to denitrification is governed by the ratio between NO3−and O2.

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