scholarly journals Analysis of erythrocyte swelling caused by osmotic shock under a non-hemolytic condition

2020 ◽  
Vol 40 (12) ◽  
pp. 23-30
Author(s):  
Kiyoshi BANDO ◽  
Ryoko OTOMO ◽  
Yukari SHIMIZU ◽  
Haruho TABUCHI
Keyword(s):  
Osmotic Shock

Properties of Isolated Mitochondria of Agaricus Bisporus: Their Relationship to Dormancy and the Germination of Spores

1973 ◽  
Vol 31 ◽  
pp. 458-459
Author(s):  
K. S. McCarty ◽  
R. F. Weave ◽  
L. Kemper ◽  
F. S. Vogel
Keyword(s):  
Mitochondrial Dna ◽  
Ethidium Bromide ◽  
Microscopic Examination ◽  
Agaricus Bisporus ◽  
Osmotic Shock ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Isolated Mitochondria ◽  
Dna Strands ◽  
Cytoplasmic Ribosomes

During the prodromal stages of sporulation in the Basidiomycete, Agaricus bisporus, mitochondria accumulate in the basidial cells, zygotes, in the gill tissues prior to entry of these mitochondria, together with two haploid nuclei and cytoplasmic ribosomes, into the exospores. The mitochondria contain prominent loci of DNA [Fig. 1]. A modified Kleinschmidt spread technique1 has been used to evaluate the DNA strands from purified whole mitochondria released by osmotic shock, mitochondrial DNA purified on CsCl gradients [density = 1.698 gms/cc], and DNA purified on ethidium bromide CsCl gradients. The DNA appeared as linear strands up to 25 u in length and circular forms 2.2-5.2 u in circumference. In specimens prepared by osmotic shock, many strands of DNA are apparently attached to membrane fragments [Fig. 2]. When mitochondria were ruptured in hypotonic sucrose and then fixed in glutaraldehyde, the ribosomes were released for electron microscopic examination.

Short-term volume and turgor regulation in yeast

2008 ◽  
Vol 45 ◽  
pp. 147-160 ◽  
Author(s):  
Jörg Schaber ◽  
Edda Klipp
Keyword(s):  
Dynamic Model ◽  
Volume Change ◽  
Volume Regulation ◽  
Time Course ◽  
Osmotic Shock ◽  
Intracellular Concentration ◽  
Short Term ◽  
Hydrodynamic Property ◽  
Turgor Regulation ◽  
Thermodynamic Framework

Volume is a highly regulated property of cells, because it critically affects intracellular concentration. In the present chapter, we focus on the short-term volume regulation in yeast as a consequence of a shift in extracellular osmotic conditions. We review a basic thermodynamic framework to model volume and solute flows. In addition, we try to select a model for turgor, which is an important hydrodynamic property, especially in walled cells. Finally, we demonstrate the validity of the presented approach by fitting the dynamic model to a time course of volume change upon osmotic shock in yeast.

scholarly journals Visualization of Chromatin in the Yeast Nucleus and Nucleolus Using Hyperosmotic Shock

2021 ◽  
Vol 22 (3) ◽  
pp. 1132
Author(s):  
Nicolas Thelen ◽  
Jean Defourny ◽  
Denis L. J. Lafontaine ◽  
Marc Thiry
Keyword(s):  
Osmotic Shock ◽  
Size Composition ◽  
Yeast Cells ◽  
Exponential Growth Phase ◽  
Cell Nuclei ◽  
Hyperosmotic Shock ◽  
Christmas Trees ◽  
Transmission Electron ◽  
Active Transcription ◽  
Decondensed Chromatin

Unlike in most eukaryotic cells, the genetic information of budding yeast in the exponential growth phase is only present in the form of decondensed chromatin, a configuration that does not allow its visualization in cell nuclei conventionally prepared for transmission electron microscopy. In this work, we studied the distribution of chromatin and its relationships to the nucleolus using different cytochemical and immunocytological approaches applied to yeast cells subjected to hyperosmotic shock. Our results show that osmotic shock induces the formation of heterochromatin patches in the nucleoplasm and intranucleolar regions of the yeast nucleus. In the nucleolus, we further revealed the presence of osmotic shock-resistant DNA in the fibrillar cords which, in places, take on a pinnate appearance reminiscent of ribosomal genes in active transcription as observed after molecular spreading (“Christmas trees”). We also identified chromatin-associated granules whose size, composition and behaviour after osmotic shock are reminiscent of that of mammalian perichromatin granules. Altogether, these data reveal that it is possible to visualize heterochromatin in yeast and suggest that the yeast nucleus displays a less-effective compartmentalized organization than that of mammals.

scholarly journals Periplasmic synthesis and purification of the human prolactin antagonist Δ1-11-G129R-hPRL

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Miriam F. Suzuki ◽  
Larissa A. Almeida ◽  
Stephanie A. Pomin ◽  
Felipe D. Silva ◽  
Renan P. Freire ◽  
...  
Keyword(s):  
Size Exclusion Chromatography ◽  
High Performance ◽  
Signal Sequence ◽  
Osmotic Shock ◽  
Size Exclusion ◽  
Specific Expression ◽  
E Coli ◽  
Human Prolactin ◽  
Exclusion Chromatography

AbstractThe human prolactin antagonist Δ1-11-G129R-hPRL is a 21.9 kDa recombinant protein with 188 amino acids that downregulates the proliferation of a variety of cells expressing prolactin receptors. Periplasmic expression of recombinant proteins in E. coli has been considered an option for obtaining a soluble and correctly folded protein, as an alternative to cytoplasmic production. The aim of this work was, therefore, to synthesize for the first time, the Δ1-11-G129R-hPRL antagonist, testing different activation temperatures and purifying it by classical chromatographic techniques. E. coli BL21(DE3) strain was transformed with a plasmid based on the pET25b( +) vector, DsbA signal sequence and the antagonist cDNA sequence. Different doses of IPTG were added, activating under different temperatures, and extracting the periplasmic fluid via osmotic shock. The best conditions were achieved by activating at 35 °C for 5 h using 0.4 mM IPTG, which gave a specific expression of 0.157 ± 0.015 μg/mL/A600 at a final optical density of 3.43 ± 0.13 A600. Purification was carried out by nickel-affinity chromatography followed by size-exclusion chromatography, quantification being performed via high-performance size-exclusion chromatography (HPSEC). The prolactin antagonist was characterized by SDS-PAGE, Western blotting, reversed-phase high-performance liquid chromatography (RP-HPLC) and MALDI-TOF–MS. The final product presented > 95% purity and its antagonistic effects were evaluated in vitro in view of potential clinical applications, including inhibition of the proliferation of cancer cells overexpressing the prolactin receptor and specific antidiabetic properties, taking also advantage of the fact that this antagonist was obtained in a soluble and correctly folded form and without an initial methionine.

scholarly journals Metallo-enzymes Released from Escherichia coli by Osmotic Shock

1968 ◽  
Vol 243 (10) ◽  
pp. 2647-2653 ◽  
Author(s):  
H F Dvorak ◽  
L A Heppel
Keyword(s):  
Escherichia Coli ◽  
Osmotic Shock

The influence of radiomimetic substances on deoxyribonucleic acid synthesis and function studied in Escherichia coli / phage systems V. A comparison of the inactivation of phages T 2, T 7 and two strains of T 4 by alkylating agents

1959 ◽  
Vol 151 (942) ◽  
pp. 148-155 ◽  
Keyword(s):  
Deoxyribonucleic Acid ◽  
Osmotic Shock ◽  
Alkylating Agents ◽  
Acid Synthesis ◽  
Acid Moiety ◽  
Rate Of Penetration ◽  
Injection Process ◽  
Deoxyribonucleic Acid Synthesis ◽  
And Function ◽  
Escherichia Coli Phage

The sensitivity of phage T 7 to epoxides and freshly prepared solutions of di(2-chloroethyl) methylamine ( HN 2) was identical with that of T 2. T 7, however, proved considerably the more sensitive to ethylenimine and to aged solutions of HN 2. It was considered that this was due to the cationic nature of these latter agents affecting the rate of penetration into the phage heads, and that the susceptibility of T 2 and resistance of T 7 to osmotic shock was a parallel phenomenon. Confirmation was afforded by the fact that a strain of T 4 sensitive to osmotic shock behaved like T 2, and a resistant strain of T 4 like T 7. These results, together with others previously reported, are believed to offer very strong evidence that inactivation of bacteriophage by alkylating agents derives from reaction with the deoxyribonucleic acid moiety, probably leading to a failure of the injection process.

The influence of selected techniques of bovine leukocyte isolation on their viability and metabolism

2011 ◽  
Vol 14 (4) ◽  
pp. 663-665 ◽  
Author(s):  
R. Urban-Chmiel ◽  
U. Lisiecka ◽  
A. Chłopaś ◽  
Ł. Kurek ◽  
A. Wernicki
Keyword(s):  
Osmotic Shock ◽  
High Viability ◽  
Reduction Assay ◽  
Isolation Methods ◽  
Pure Cell ◽  
Nbt Reduction ◽  
Cell Fractions ◽  
Number Of Cells

The influence of selected techniques of bovine leukocyte isolation on their viability and metabolism The aim of the study was to assess the effect of selected isolation methods on the viability and metabolism of bovine leukocytes. The cells were isolated using a Ficoll 1077, Histopaque 1083 gradient and osmotic shock method, and Ficoll or Histopaque with osmotic shock. Evaluation were made of the total number of cells, viability after isolation and in 24h culture on RPMI 1640 medium and metabolism with NBT reduction assay. Microscopic and cytometric evaluation of the leukocytes revealed that the isolation methods applied had an influence on their number and viability. Based on the results it can be concluded that isolation methods of cells in a Histopaque or Ficoll yield highly pure cell fractions with high viability.

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