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91.
Induction characteristics of oxygen regulated proteins 总被引:14,自引:0,他引:14
C S Heacock R M Sutherland 《International journal of radiation oncology, biology, physics》1986,12(8):1287-1290
Extreme hypoxia induces many changes in the biology of cells, including the enhanced synthesis of oxygen regulated proteins (ORPs). We investigated the conditions required for the induction of ORPs and by modifying culture conditions, eliminated variables other than oxygen concentration. Several exponentially growing rodent and human cell lines were examined before, during, and after various periods of extreme hypoxia. The following responses were analyzed: cell growth, clonogenic survival, glucose consumption, lactate production, media pH, total protein synthesis, and specific protein synthesis. EMT6/Ro cells did not increase in cell number or progress through the cell cycle after initiation of extreme hypoxia. Cell morphology and cell survival were nearly normal for up to 12 hr of hypoxia. During this period, media pH remained constant, with the concentrations of glucose and lactate being virtually indistinguishable from aerobic cultures or initial values. Associated with these conditions, a marked inhibition of total protein synthesis was observed for EMT6/Ro cells, such that the hypoxic protein synthesis rate was about 60% of the aerobic rate. However, enhanced synthesis of a set of proteins, designated as ORPs, was preferentially induced in less than 6 hr. The molecular weights of the five major ORPs are 260, 150, 100, 80 and 33 kD. Under these conditions, the primary inducing agent was a low concentration of oxygen. This set of ORPs was distinctly different from the set of heat induced (heat-shock) proteins, but included the major 100 kD and 80 kD glucose regulated proteins. Although the functions of ORPs are unknown, their induction under conditions that are known to modify the sensitivity of cancer cells to therapeutic agents suggests that the presence of ORPs should be further investigated to determine their possible value in diagnosis and predicting treatment response. 相似文献
92.
R M Sutherland B Sordat J Bamat H Gabbert B Bourrat W Mueller-Klieser 《Cancer research》1986,46(10):5320-5329
Oxygenation and development of necrosis were evaluated in multicellular spheroids of poorly differentiated (HT29) and moderately well-differentiated (Co112) human adenocarcinoma of the colon. Spheroids were grown in vitro under well-controlled oxygen and nutrient conditions in spinner flasks up to sizes of 2800-micron diameter after 5 wk of culture. Morphological studies showed that the Co112 spheroids contained pseudoglandular structures with lumen, very similar to the characteristics of the original tumor specimen from the patient and to the cells when grown as xenograft tumors in nude mice. Microelectrodes were used to measure the oxygen tension (PO2) profile within individual spheroids at different stages of growth. Histological sections through the centers of spheroids were measured to determine the thickness of the viable rim of cells surrounding spheroid necrotic centers in order to estimate the size of the severely hypoxic zone of cells by comparison with the PO2 profiles of the same spheroids. The data demonstrate significant differences between these two human colon tumor spheroid systems. Both spheroid types exhibited steep PO2 gradients at relatively small sizes of less than 600-micron diameter, but for any given size in this range, the more differentiated Co112 spheroids were more hypoxic. Although severe hypoxia (PO2, less than 10 mm of Hg) was present in both spheroid types at larger sizes, there was a significant difference in the central PO2 values which were between 5 and 10 mm of Hg in large Co112 spheroids but remained at or close to 0 mm of Hg in large HT29 poorly differentiated human colon tumor spheroids. The presence of pseudoglandular structures and lumen in the Co112 spheroids was associated with changes in the shape of PO2 profiles. Such profiles have not previously been seen in other poorly differentiated human or rodent tumor spheroids. Furthermore, the PO2 profiles of both of these human tumor spheroid types were often continuously curving with a very shallow gradient in the inner edge of the viable rim of cells surrounding the necrotic center. Regulation of oxygen consumption and/or diffusion in these inner regions of human spheroids could produce these continuously curving PO2 gradients. 相似文献
93.
Determination of malondialdehyde-induced DNA damage in human tissues using an immunoslot blot assay 总被引:3,自引:4,他引:3
Leuratti C; Singh R; Lagneau C; Farmer PB; Plastaras JP; Marnett LJ; Shuker DE 《Carcinogenesis》1998,19(11):1919-1924
Malondialdehyde (MDA) is a product of lipid peroxidation and prostaglandin
biosynthesis. It is mutagenic and carcinogenic and the major adduct formed
by reaction with DNA, a highly fluorescent pyrimidopurinone (M1-dG), has
been detected in healthy human liver and leukocyte DNA. Analytical methods
used so far for the detection of M1- dG have not been applied to a large
number of individuals or variety of samples. Often, only a few microg of
DNA from human tissues are available for analysis and a very sensitive
assay is needed in order to detect background levels of M1-dG in very small
amounts of DNA. In this paper, the development of an immunoslot blot (ISB)
assay for the measurement of MI-dG in 1 microg of DNA is described. The
limit of detection of the assay is 2.5 adducts per 10(8) bases. A series of
human samples were analysed and levels of 5.6-9.5 (n = 8) and 3.1-64.3 (n =
42) of M1-dG per 10(8) normal bases were detected in white blood cell and
gastric biopsy DNA, respectively. Results on four human samples were
compared with those obtained using an HPLC/32P-post- labelling (HPLC/PPL)
method previously developed and indicated a high correlation between M1-dG
levels measured by the two assays. The advantages of ISB over other assays
including HPLC/PPL, such as the possibility of analysing 1 microg
DNA/sample and the fact that it is less time-consuming and laborious, means
that it can be more easily used for routine analysis of a large number of
samples in biomonitoring studies.
相似文献
94.
Dixon B. Kaufman George W. Burke III David S. Bruce Christopher P. Johnson A. Osama Gaber David E. R. Sutherland Robert M. Merion Scott A. Gruber Eugene Schweitzer John P. Leone Christopher L. Marsh Edward Alfrey Waldo Concepcion Mark D. Stegall James A. Schulak Paul F. Gores Enrico Benedetti Craig Smith Alice K. Henning Fernando Kuehnel Sarah King William E. Fitzsimmons 《American journal of transplantation》2003,3(7):855-864
A randomized, multicenter, prospective study was conducted at 18 pancreas transplant centers in the United States to determine the role of induction therapy in simultaneous pancreas-kidney (SPK) transplantation. One hundred and 74 recipients were enrolled: 87 recipients each in the induction and noninduction treatment arms. Maintenance immunosuppression consisted of tacrolimus, mycophenolate mofetil, and corticosteroids. There were no statistically significant differences between treatment groups for patient, kidney, and pancreas graft survival at 1-year. The 1-year cumulative incidence of any treated biopsy-confirmed or presumptive rejection episodes (kidney or pancreas) in the induction and noninduction treatment arms was 24.6% and 31.2% (p = 0.28), respectively. The 1-year cumulative incidence of biopsy-confirmed, treated, acute kidney allograft rejection in the induction and noninduction treatment arms was 13.1% and 23.0% (p = 0.08), respectively. Biopsy-confirmed kidney allograft rejection occurred later post-transplant and appeared to be less severe among recipients that received induction therapy. The highest rate of Cytomegalovirus (CMV) viremia/syndrome was observed in the subgroup of recipients who received T-cell depleting antibody induction and received organs from CMV serologically positive donors. Decisions regarding the routine use of induction therapy in SPK transplantation must take into consideration its differential effects on risk of rejection and infection. 相似文献
95.
为了寻找毒性低、增敏作用强的乏氧细胞放射增敏剂,设计并合成了一系列5-溴-,5-甲基-,和5-未取代的3-硝基-1,2,4-三唑-1-乙酰胺类化合物,用HeLaS3细胞进行了体外试验。结果表明5-溴取代衍生物的增敏作用强于相应的5-甲基-或5-未取代的硝基三唑衍生物,但是它们的毒性亦增大。修饰1位乙酰胺侧链也可以改变化合物的增敏作用和亲脂性。在所测定的化合物中TA-101[2-(3-硝基-1-三唑基)乙酰胺]由于有高的增敏作用和低亲脂性,可能是一个有希望的放射增敏剂。 相似文献
96.
Experimental assessment of a new research tool for the estimation of two-dimensional myocardial strain 总被引:5,自引:0,他引:5
Langeland S Wouters PF Claus P Leather HA Bijnens B Sutherland GR Rademakers FE D'hooge J 《Ultrasound in medicine & biology》2006,32(10):1509-1513
One-dimensional strain imaging has been shown to be angle dependent. To address this problem, a new methodology, 2D-strain, has become available. The aim of this study was to validate this methodology in an in vivo set-up against sonomicrometry. In five open chest sheep, ultrasound gray-scale images were acquired of the inferolateral wall from two different angles. The longitudinal and radial strain components were simultaneously extracted using the novel 2D-strain methodology. The extracted values were compared with sonomicrometry using Bland-Altman statistics and correlation coefficients. A good agreement was found for the longitudinal strain component, while, for the radial strain estimates, the accuracy was less. 2D-strain is a fast and accurate tool to assess longitudinal strain from apical views. Further improvements are needed for the method to be sufficiently accurate in estimating the deformation perpendicular or close to perpendicular to the ultrasound beam. 相似文献
97.
98.
Insertion of an internal metal prosthesis to replace a deficient superior acetabular margin in subluxating hips is a feasible and practical procedure. Such an operation can be more simple, more efficient, and less shocking than the conventional procedure. 相似文献
99.
100.