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Effects of pchloromercuribrnzoate and n-ethylmaleimide on hemolysis under hydrostatic pressure of human erythocytes

100%

Yamaguchi T. , Terada S.

Polish Hyperbaric Research

2005

tom Nr 1(10)

44-43

The relation between high-pressure (200mpa)-induced hemolisis and spectrin tetramer-dimer equilibrium was examined by using human erythrocytes treatedwith N-ethylmaleimide (NEM) and p-chloromercuribenzoate (PCMB). The values of hemolysis at 200 Mpa of NEM-treated erythrocytes increased upon mild modification of membrane SH groups, but decreased upon severe modification. In both cases, the membrane structure of NEM-treated cells became more stable against high pressure upon further incubation at 370C in reagent-free buffer. In PCMB-treated erythrocytes, the cells were hemolyzed completely at 200 Mpa. On the other hand, the concentration of spectrin dimers increased upon treatment with NEM, but remained constant for further incubation at 370C. In the case of PCMB-treated cells, spectrin tetramers did not dissociate into dimers as with NEM. These results suggest that hemolytic properties at 200 Mpa in erytrocytes treated with SH-reactive reagents are inexplicable in terms of the dissociation only of spectrin.

Analysis of high-pressure-induced disruption of human erythrocytes by flow cytometry

100%

Yamaguchi T. , Terada S.

Cellular and Molecular Biology Letters

2003

tom 08

nr 4

High-pressure-induced hemolysis is suppressed by pretreating human erythrocytes at 49°C, or enhanced by pretreatment with trypsin. So, the response of these pretreated cells to a pressure of 200 MPa was examined using flow cytometry. In the case of intact erythrocytes, a major product was fragmented particles. From 49°C-pretreated cells, vesicles were mainly released. Trypsin-pretreated cells mainly produced open ghosts. Additionally, intact erythrocytes, 49°C-pretreated ones, and trypsin-pretreated ones also released at 200 MPa vesicles of diameter 464 ± 9, 259 ± 18, and 574 ± 16 nm, respectively. These results suggest that mother cells, fragmented particles, vesicles, and open ghosts from 200 MPa-treated erythrocytes are easily monitored by flow cytometry and that the size of released vesicles may also be an important factor in high-pressure-induced hemolysis.

DNA replication reaction in Xenopus cell-free system is suppressed by high pressure

70%

Takahashi H. , Yamaguchi T. , Koga M. , Kageura H. , Terada S.

Cellular and Molecular Biology Letters

2004

tom 09

nr 3

Previously, we demonstrated that when mouse erythroleukemia cells are exposed to a pressure of 80 MPa, the cell-cycle progression of S-phase cells is retarded. To examine the effects of high pressure on DNA replication, we used a Xenopus cell-free system. From cell-cycle progression of sperm nuclei, it was found that sperm nuclei are stable to a pressure of 80 MPa, whereas egg extracts are susceptible to high pressure. Similarly, biotin-16-dUTP was incorporated into 80 MPa-treated sperm nuclei in pressure-untreated extracts, but not into naive sperm nuclei in 80 MPa-treated extracts. These results indicate that DNA replication in Xenopus cell-free system is suppressed by the susceptibility of the extracts to a pressure of 80 MPa.