Robust Analytical Methods for the Accurate Quantification of the Total Biomass Composition of Mammalian Cells

AbstractOcean warming and the increased prevalence of coral bleaching events threaten coral reefs. However, the biology of corals during and following bleaching events under field conditions is poorly understood. We examined bleaching and post-bleaching recovery inMontipora capitataandPorites compressacorals that either bleached or did not bleach during a 2014 bleaching event at three reef locations in Kāne‘ohe Bay, O‘ahu. We measured changes in chlorophylls, biomass, and nutritional plasticity using stable isotopes (δ13C, δ15N). Coral traits showed significant variation among bleaching conditions, reef sites, time periods, and their interactions. Bleached colonies of both species had lower chlorophyll and total biomass. WhileM. capitatachlorophyll and biomass recovered three months later,P. compressachlorophyll recovery was location-dependent and total biomass of previously bleached colonies remained low. Biomass energy reserves were not affected by bleaching, insteadM. capitataproteins andP. compressabiomass energy declined over time, andP. compressalipid biomass was site-specific. Stable isotope analyses of host and symbiont tissues did not indicate increased heterotrophic nutrition in bleached colonies of either species, during or after thermal stress. Instead, mass balance calculations revealed variance in δ13C values was best explained by augmented biomass composition, whereas δ15N values reflected spatial and temporal variability in nitrogen sources in addition to bleaching effects on symbiont nitrogen demand. These results emphasize total biomass quantity may change substantially during bleaching and recovery. Consequently, there is a need to consider the influence of biomass composition in the interpretation of isotopic values in corals.

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Biomass yield in production systems of soybean sown in succession to annual crops and cover crops

Pesquisa Agropecuária Brasileira ◽

10.1590/s0100-204x2017000800003 ◽

2017 ◽

Vol 52 (8) ◽

pp. 582-591 ◽

Cited By ~ 3

Author(s):

Leandro Pereira Pacheco ◽

Andressa Selestina Dalla Côrt São Miguel ◽

Rayane Gabriel da Silva ◽

Edicarlos Damacena de Souza ◽

Fabiano André Petter ◽

...

Keyword(s):

Cover Crops ◽

Production Systems ◽

Pennisetum Glaucum ◽

Block Design ◽

Biomass Composition ◽

Total Biomass ◽

Crop Season ◽

Yield Increase ◽

Annual Crops ◽

Tillage System

Abstract: The objective of this work was to evaluate the biomass (leaves and stems) production of annual and cover crops sown as second crop, and its effects on soybean yield in succession. The experiment was carried out in the 2014/2015 and 2015/2016 crop seasons. Soybean was sown in the crop season and in the second crop, in a randomized complete block design, in nine production systems (treatments) consisting of annual crops (corn, sunflower, and cowpea) and cover crops (Pennisetum glaucum, Crotalaria breviflora, C. spectabilis, Urochloa ruziziensis, Cajanus cajan, Stylosanthes sp., and U. brizantha), which were grown in monocropping or intercropping systems, besides fallow as a control. Monocropped P. glaucum and U. ruziziensis showed a faster establishment and growth of plants, higher-total biomass and soil cover rate in the 2014 crop season. In 2015, corn intercropped with U.ruziziensis and C.spectabilis, and sunflower with U.ruziziensis stood out for total biomass production during flowering and after harvesting of corn and sunflower grains. Biomass composition in the systems showed greater proportions of stems than of leaves, and C.spectabilis stood out after senescence. Sown as a second crop, C. spectabilis promotes yield increase of soybean grown in succession in the no-tillage system.

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Biomass, composition and temporal dynamics of soil organisms of a silt loam soil under conventional and integrated management.

Netherlands Journal of Agricultural Science ◽

10.18174/njas.v38i3a.16589 ◽

1990 ◽

Vol 38 (3A) ◽

pp. 283-302 ◽

Cited By ~ 6

Author(s):

L. Brussaard ◽

L.A. Bouwman ◽

M. Geurs ◽

J. Hassink ◽

K.B. Zwart

Keyword(s):

Temporal Dynamics ◽

Integrated Management ◽

Biomass Composition ◽

Total Biomass ◽

Soil Organisms ◽

N Transfer ◽

Loam Soil ◽

Flow Through ◽

C And N ◽

Main Food Source

Field plots (Netherlands) under conventional (CF) or integrated (IF) farming and cropped to winter wheat were sampled in 1986 for various soil organisms. Organisms were assembled in functional groups, based on their main food source and life-history characteristics. Total biomass of soil organisms was, on average, 690 kg C/ha under CF and 907 kg C/ha under IF during the growing season. Bacteria constituted > 90%, fungi ~ 5%, and protozoa < 2% of the total biomass. Nematodes and microarthropods were less important in terms of biomass C. Carbon flow through the protozoa annually was estimated to be 158 and 195 kg C/ha in CF and IF, resp., corresponding to 20% of the estimated bacterial production in both CF and IF. Nitrogen mineralization by the protozoa annually was estimated to be 30.5 and 37.6 kg N/ha in CF and IF, resp. Nematodes were less important than protozoa in terms of direct C and N transfer. The direct contribution from microarthropods was insignificant. Results are discussed in terms of effects of the soil biota, in particular the soil fauna, on C and N transfer in arable soil. (Abstract retrieved from CAB Abstracts by CABI’s permission)

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Analytical methods for the quantification of cisplatin, carboplatin, and oxaliplatin in various matrices over the last two decades

Current Pharmaceutical Analysis ◽

10.2174/1573412918666210929105058 ◽

2021 ◽

Vol 18 ◽

Author(s):

Hajer Hrichi ◽

Noura Kouki ◽

Haja Tar

Keyword(s):

Analytical Methods ◽

Biological Fluids ◽

Pharmaceutical Formulations ◽

Atomic Absorption Spectrophotometry ◽

Literature Reviews ◽

Small Doses ◽

Inherent Problem ◽

Uv Visible ◽

Accurate Quantification

Background: Platinum derivatives including cisplatin and its later generations carboplatin, and oxaliplatin remain the most largely used drugs in the therapy of malignant diseases. They exert notable anticancer activity towards numerous types of solid tumors such as gastric, colorectal, bladder, ovary, and several others. The chemotherapeutic activity of these compounds, however, is associated with many unwanted side effects and drug resistance problems limiting their application and effectiveness. Proper dosage is still an inherent problem, as these drugs are usually prescribed in small doses. Objective: Several analytical methods have been reported for the accurate quantification of cisplatin, carboplatin, and oxaliplatin and their metabolites either alone or in combination with other chemotherapeutic drugs, in different matrices such as pharmaceutical formulations, biological fluids, cancer cells, and environmental samples. The main goal of this review is to systematically study the analytical methods already used for the analysis of cisplatin, carboplatin, and oxaliplatin in various matrices during the last two decades. Results and Conclusion: In the literature, reviews showed that numerous analytical methods such as electroanalytical, UV-visible spectrophotometry, chromatographic, fluorescence, atomic absorption spectrophotometry, and other spectroscopic methods combined with mass spectrometry were used for the determination of these compounds in various matrices.

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Photorhabdus Luminescens Toxin complex (TCC) a recombinant injection nano-machine – delivery of recombinant Yersinia enterocolitica YopT

10.1101/698027 ◽

2019 ◽

Author(s):

Peter Ng’ang’a ◽

Julia K. Ebner ◽

Matthias Plessner ◽

Klaus Aktories ◽

Gudula Schmidt

Keyword(s):

Mammalian Cells ◽

Photorhabdus Luminescens ◽

Therapeutic Approaches ◽

Highly Active ◽

Biological Studies ◽

Toxin Complex ◽

Protein Toxins ◽

Protein Transporters ◽

Accurate Quantification ◽

Proteins And Peptides

ABSTRACTEngineering delivery systems for proteins and peptides into mammalian cells is an ongoing challenge for cell biological studies as well as for therapeutic approaches. Photorhabdus luminescens toxin complex (PTC) is a heterotrimeric protein complex able to deliver diverse protein toxins into mammalian cells. We engineered the syringe like nano-machine for delivery of protein toxins from different species. Additionally, we loaded the highly active copepod luciferase Metridia longa M-Luc7 for accurate quantification of injected molecules. We suggest that besides the size also the charge of the cargo defines the efficiency of packing and transport into mammalian cells. Our data show that the Photorhabdus luminescens toxin complex constitutes a powerful system to inject recombinant proteins, peptides and potentially other molecules like aptamers into mammalian cells. In contrast to other protein transporters based on pore formation, the cargo is protected from degradation. The system opens new perspectives for cell research and pharmacology.

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Ultrastructural Alterations in Hela Cells Induced by Spider Venom

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100060118 ◽

1967 ◽

Vol 25 ◽

pp. 20-21

Author(s):

Dale E. McClendon ◽

Paul N. Morgan ◽

Bernard L. Soloff

Keyword(s):

Growth Medium ◽

Mammalian Cells ◽

Carcinoma Cells ◽

Venom Protein ◽

Sterile Saline ◽

Carbon Coated ◽

Available Information ◽

Loxosceles Reclusa ◽

Essential Growth ◽

Solution Cultures

It has been observed that minute amounts of venom from the brown recluse spider, Loxosceles reclusa, are capable of producing cytotoxic changes in cultures of certain mammalian cells (Morgan and Felton, 1965). Since there is little available information concerning the effect of venoms on susceptible cells, we have attempted to characterize, at the electron microscope level, the cytotoxic changes produced by the venom of this spider.Cultures of human epithelial carcinoma cells, strain HeLa, were initiated on sterile, carbon coated coverslips contained in Leighton tubes. Each culture was seeded with approximately 1x105 cells contained in 1.5 ml of a modified Eagle's minimum essential growth medium prepared in Hank's balanced salt solution. Cultures were incubated at 36° C. for three days prior to the addition of venom. The venom was collected from female brown recluse spiders and diluted in sterile saline. Protein determinations on the venom-were made according to the spectrophotometric method of Waddell (1956). Approximately 10 μg venom protein per ml of fresh medium was added to each culture after discarding the old growth medium. Control cultures were treated similarly, except that no venom was added. All cultures were reincubated at 36° C.

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Use of Uranium-Labeled Antibodies for Electron Microscopic Localization of Various Antigens in Mammalian Cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100060696 ◽

1967 ◽

Vol 25 ◽

pp. 136-137

Author(s):

J. P. Petrali ◽

E. J. Donati ◽

L. A. Sternberger

Keyword(s):

Endoplasmic Reticulum ◽

Hela Cells ◽

Mammalian Cells ◽

Specific Antiserum ◽

Electron Microscopic ◽

E Coli ◽

Cytoplasmic Membranes ◽

Viral Progeny ◽

Ultrathin Sections ◽

Electron Microscopic Localization

Specific contrast is conferred to subcellular antigen by applying purified antibodies, exhaustively labeled with uranium under immunospecific protection, to ultrathin sections. Use of Seligman’s principle of bridging osmium to metal via thiocarbohydrazide (TCH) intensifies specific contrast. Ultrathin sections of osmium-fixed materials were stained on the grid by application of 1) thiosemicarbazide (TSC), 2) unlabeled specific antiserum, 3) uranium-labeled anti-antibody and 4) TCH followed by reosmication. Antigens to be localized consisted of vaccinia antigen in infected HeLa cells, lysozyme in monocytes of patients with monocytic or monomyelocytic leukemia, and fibrinogen in the platelets of these leukemic patients. Control sections were stained with non-specific antiserum (E. coli).In the vaccinia-HeLa system, antigen was localized from 1 to 3 hours following infection, and was confined to degrading virus, the inner walls of numerous organelles, and other structures in cytoplasmic foci. Surrounding architecture and cellular mitochondria were unstained. 8 to 14 hours after infection, antigen was localized on the outer walls of the viral progeny, on cytoplasmic membranes, and free in the cytoplasm. Staining of endoplasmic reticulum was intense and focal early, and weak and diffuse late in infection.

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Ultrastructural Changes in Three Different Mammalian Cells Following Ultraviolet Laser Irradiation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100095017 ◽

1972 ◽

Vol 30 ◽

pp. 350-351

Author(s):

K. Shankar Narayan ◽

Kailash C. Gupta ◽

Tohru Okigaki

Keyword(s):

Ultraviolet Light ◽

Mammalian Cells ◽

Biological Effects ◽

Survival Rates ◽

Chinese Hamster ◽

Cell Types ◽

Differential Sensitivity ◽

Ultrastructural Changes ◽

Ultraviolet Laser

The biological effects of short-wave ultraviolet light has generally been described in terms of changes in cell growth or survival rates and production of chromosomal aberrations. Ultrastructural changes following exposure of cells to ultraviolet light, particularly at 265 nm, have not been reported.We have developed a means of irradiating populations of cells grown in vitro to a monochromatic ultraviolet laser beam at a wavelength of 265 nm based on the method of Johnson. The cell types studies were: i) WI-38, a human diploid fibroblast; ii) CMP, a human adenocarcinoma cell line; and iii) Don C-II, a Chinese hamster fibroblast cell strain. The cells were exposed either in situ or in suspension to the ultraviolet laser (UVL) beam. Irradiated cell populations were studied either "immediately" or following growth for 1-8 days after irradiation.Differential sensitivity, as measured by survival rates were observed in the three cell types studied. Pattern of ultrastructural changes were also different in the three cell types.

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Stereo modeling of HVEM specimens by serial tilting and computer graphics

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100141913 ◽

1986 ◽

Vol 44 ◽

pp. 34-35

Author(s):

J.R. McIntosh ◽

D.L. Stemple ◽

William Bishop ◽

G.W. Hannaway

Keyword(s):

Computer Graphics ◽

Analytical Methods ◽

Photographic Film ◽

Complex Structures ◽

Multiple Objects ◽

Multiple Images ◽

3 Dimensional

EM specimens often contain 3-dimensional information that is lost during micrography on a single photographic film. Two images of one specimen at appropriate orientations give a stereo view, but complex structures composed of multiple objects of graded density that superimpose in each projection are often difficult to decipher in stereo. Several analytical methods for 3-D reconstruction from multiple images of a serially tilted specimen are available, but they are all time-consuming and computationally intense.

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