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I J Spiro

Publications and source records attributed to I J Spiro.

36 records · Page 2Linked to original sources
PubMed1991

Sensitization of low-dose-rate irradiation by nonlethal hyperthermia.

To assess whether hyperthermia could radiosensitize cells irradiated at a low dose rate, Chinese hamster V79 cells were simultaneously heated and irradiated at 0.86 Gy/h. The data showed that heat treatments at 39 and 40 degrees C, which did not induce heat killing alone or high-dose-rate radiosensitization, resulted in enhanced cell killing with low-dose-rate irradiation. The dose-modification factor (ratio of the slopes of the curves for low dose rate and high dose rate) was reduced to 1.8 at 39 degrees C and 1.4 at 40 degrees C, compared to a value of 2.1 at 37 degrees C. These data indicate that nonlethal heat treatments can cause enhanced radiosensitization under low-dose-rate conditions. The implications of these results for interstitial thermoradiotherapy are discussed.

Animals↗
PubMed1987

The effect of hypoxic cell sensitizers at different irradiation dose rates.

The effect of 0.1-5 mM misonidazole and SR 2508 on hypoxic V79 cellular survival at acute (498 cGy/min) and low (890 and 933 cGy/h) irradiation dose rates was measured and compared. The experiments were designed to delineate the oxygen mimetic phenomenon and the preincubation effect of these chemicals at these dose rates. Linear regression analysis of the survival data in terms of the linear quadratic model yielded values of alpha and beta. In the absence of drug, the linear coefficient was independent of dose rate, whereas the quadratic term was greatly reduced at low dose rate. At all dose rates, the preincubation effect affected primarily the alpha term, with little influence on beta. In contrast, the oxygen mimetic phenomenon predominantly affected the beta term. Overall, the radiosensitizing ability of these drugs was higher at low dose rate than at acute dose rate.

Animals↗
PubMed1986

Transfection of a human gene for the repair of X-ray- and EMS-induced DNA damage.

EM9 cells are a line of Chinese hamster ovary cells that are sensitive to killing by ethylmethanesulfonate (EMS) and X ray, since they are unable to repair the DNA damage inflicted by these agents. Through DNA-mediated gene transfer, human DNA and a selectable marker gene, pSV2neo, were transfected into EM9 cells. Resistant clones of transfected cells were selected for by growth in EMS and G418 (an antibiotic lethal to mammalian cells not containing the transfected neo gene). One primary clone (APEX1) and one secondary clone (TEMS2) were shown to contain both marker and human DNA sequences by Southern blot. In cell survival studies, APEX1 was shown to be as resistant to EMS and X ray as the parental cell type AA8 (CHO cells). TEMS2 cells were found to be partially resistant to EMS and X ray, displaying an intermediate phenotype more sensitive than AA8 cells but more resistant than EM9 cells. Alkaline elution was used to assess the DNA strand-break rejoining ability of these cells at 23 degrees C. APEX1 cells showed DNA repair capacity equal to that of AA8 cells; 75% of the strand breaks were repaired with a rejoining T 1/2 of 3 min. TEMS2 showed similar levels of repair but a T 1/2 for repair of 9 min. EM9 cells repaired only 25% of the breaks and showed a T 1/2 for repair of 16 min. The DNA repair data are consistent with the survival data in that the more resistant cell lines showed a greater capacity for DNA repair. The data support the conclusion that APEX1 and TEMS2 cells contain a human DNA repair gene.

Animals↗
PubMed1985

Detection of S-phase overreplication following chronic hypoxia using a monoclonal anti-BrdUrd.

We have examined cytokinetic perturbations induced in Chinese hamster V-79 cells in vitro during and following exposure to chronic hypoxia employing simultaneous flow cytometric measurement of incorporated BrdUrd and DNA content. These data indicate hypoxia inhibited G1 progression into S-phase, but did not significantly delay G2M division and progression into G1. Also, upon reaeration after 20 hr in hypoxia, cells originally in G1 exhibited significant kinetic delay. BrdUrd pulse/chase and pulse/fix data indicated DNA replication was reduced, but not completely inhibited during hypoxia. Also, between 6 and 20 hr of chronic hypoxia and following reaeration, a subset of the original S-phase cells overreplicated their DNA, such that these cells had greater than 2C DNA content. This subpopulation was estimated on the average to comprise approximately 20% of the total population (30% of the treated S-phase subpopulation) by 24 hr following reaeration after 20 hr hypoxia. These results are discussed in light of the similarities between overreplication and gene amplification observed under certain conditions with other agents, which like chronic hypoxia, are used to transiently disrupt DNA synthesis.

Animals↗
PubMed1985

The variation of OER with dose rate.

The oxygen enhancement ratio (OER) has been measured as a function of dose rate from 276 Gy/hr to 0.89 Gy/hr for V-79 cells irradiated at 23 degrees C or 37 degrees C. As dose rate is decreased, the OER initially increases, from a value of 3.0, to a maximum value of 3.7 to 4.0, at a dose rate between 20 and 60 Gy/hr. The OER subsequently decreases with further dose rate reduction to a minimum value of 2.4 at the lowest dose rate. Similar experiments conducted with cells in nutrient deprived conditions exhibited a monotonic decrease in OER from 3.0 to 1.7 with dose rate reduction. These experimental findings can be understood in terms of the sublethal damage repair capability of cells under different pO2 and nutrient conditions.

Animals↗
PubMed1985

Oxygen radiosensitisation at low dose rate.

Oxygen radiosensitisation has been studied at dose rates of 600, 3.37 and 0.89 Gy/h at pO2 levels of 0.001, 0.03, 0.1, 0.3, 1, 3, 10 and 21% in the gas phase. The oxygen enhancement ratio (OER), evaluated at 2% cellular survival, exhibited a decrease with dose rate from a value of 3.2 at the acute dose rate, to 2.4 at the lowest dose rate. This observation results from a decreased dose-rate effect on hypoxic cells, which is attributed to the partial suppression of sublethal damage (SLD) repair under hypoxic conditions. Oxygen radiosensitisation at the acute dose rate agrees with the calculated values based on the oxygen fixation hypothesis. Direct application of the Howard-Flanders and Moore equation to results obtained at low dose rate is not appropriate due to the influence of pO2 on SLD repair which affects radiosensitivity at low dose rate. When cells are irradiated at 3.37 Gy/h under nutrient-deprived condition (i.e. in Hanks balanced salt solution without glucose), low levels of oxygen appear to be more radioprotective than extreme hypoxia. Specifically, cells irradiated with 0.03% and 0.1% O2 are more radioresistant than cells under N2, with enhancement factors of 0.7 and 0.8, respectively. This phenomenon can be understood in terms of the ability of moderately hypoxic cells (0.03%-0.1% O2), and inability of anoxic cells, to repair SLD under nutrient deprived conditions. Radiosensitisation by these low levels of oxygen is insufficient to offset the difference caused by the disparate SLD repair capabilities.

Animals↗
PubMed1985

Cellular and molecular repair of X-ray-induced damage: dependence on oxygen tension and nutritional status.

Cellular and molecular repair was studied at 23 degrees C using split-dose recovery and alkaline elution techniques, respectively, as a function of cellular oxygen and nutrient conditions. Hypoxic cells (0.001% O2) in full medium showed a partial reduction in the level of sublethal damage (SLD) repair relative to aerated cells (21% O2, OER equal to 3.2 relative to 0.001% O2); the respective repair kinetics were similar with a common repair half-time of 30 min. Similarly, hypoxic cells showed a slight reduction in strand break rejoining capacity compared to aerated cells. Under nutrient deprivation, anoxic cells displayed no SLD repair or strand break repair, while aerated cells exhibited the same level of SLD and strand break repair as for well-fed cells. In addition, nutrient deprived cells at low O2 levels (0.03%, OER equal to 1.15) displayed normal SLD and strand break repair capability. These results indicate that both nutrient and O2 deprivation are necessary for complete inhibition of cellular and molecular repair, and low levels of O2 (0.03%) can effectively reverse this inhibition.

Animals↗
PubMed1985

The effects of cellular glutathione elevation on the oxygen enhancement ratio.

The radiation responses at various oxygen tensions were evaluated in V79 Chinese hamster cells under conditions where their nonprotein thiols, primarily glutathione (GSH), were elevated by 2-oxothiazolidine-4-carboxylate (OTZ). OTZ, when cleaved by intracellular oxoprolinase, provides the cell with cysteine which stimulates GSH synthesis. A 2-hr pretreatment with 10 mM OTZ elevated GSH to 200% of controls. This elevation in GSH offered no protection to aerated cells; however, for O2 tensions less than or equal to 40,000 ppm modest protection was observed as evidenced by an increase in oxygen enhancement ratio. GSH elevation afforded maximal protection between 1000 and 10,000 ppm O2; however, the extent of protection was relatively small (protection factor = 1.3).

Animals↗
PubMed1984

Cell killing, radiosensitization and cell cycle redistribution induced by chronic hypoxia.

Some of the biological changes associated with extreme hypoxia at 37 degrees C (less than 10 ppm pO2) were examined in Chinese hamster V79 cells. Specifically, extreme hypoxia caused an initial decrease in plating efficiency to 55% in 4 hr after the onset of hypoxia. Beyond this time, the decline in plating efficiency was more gradual reaching 35% of control at 20 hr. Flow microfluorimetry (FMF) studies, in which cells are sorted on the basis of DNA content and then assayed for viability, demonstrated that mid S phase cells were most sensitive to chronic hypoxia, with surviving fraction equal to 2.5% at 20 hr. Furthermore, the viability of G1 and G2/M cells, after 20 hr of hypoxic storage, was also reduced to 20 and 7.6%, respectively. Hypoxia also caused alterations in the cell cycle distribution of initially asynchronous cells, as determined by dual parameter FMF measurements of both cellular DNA content and incorporated BudR. In particular, G2/M cells completed mitosis, while G1 cells showed little or no movement. Lastly, cells stored in chronic hypoxia displayed an enhanced radiosensitivity when compared to acutely hypoxic cells. Possible reasons for these observations are discussed.

Animals↗
PubMed1984

Dose-rate effect between 1 and 10 Gy/min in mammalian cell culture.

A dose-rate effect is observed between 1 and 10 Gy/min for cultured Chinese hamster V-79 fibroblast cells irradiated under pO2 levels of less than 10 ppm, 100 ppm, 300 ppm and 21%. The dose for a cellular surviving fraction of 0.1% increases by 10-15% as the dose rate is changed from 10 to 1 Gy/min. This observed dose-rate effect is consistent with the repair of sublethal damage taking place during the radiation delivery. A related finding is that acute hypoxia in cell culture does not inhibit sublethal damage repair.

Animals↗
PubMed1983

Physical dosimetry of 125I seeds of a new design for interstitial implant.

The physical characteristics of a new 125I seed, consisting of radioactive iodine absorbed on a silver wire and contained in a sealed titanium shell, have been measured. Advantages of the new seed design are: increased radiopacity, possible determination of seed orientation in an implant for dosimetric calculations, and source strength specification traceable to the National Bureau of Standards. Spectroscopic analysis of the new seed using an intrinsic Ge detector revealed the 27.4, 31.4 and 35.5kev photons from the decay of 125I, and in addition, 22.1 and 25.2kev fluorescent X ray from the silver wire. Measured and calculated relative dose distribution along the perpendicular bisector of the new seed is similar to that of the existing seed, with a slightly more rapid fall-off due to the existence of the lower energy photons. The measured angular distributions of the seeds of the two designs are similar, exhibiting significant anisotropy. A protocol of source strength specification, choice of effective gamma constant value and dose calculation relative to 125I implants is suggested.

Brachytherapy↗
PubMed1983

Effect of hyperthermia on isolated DNA polymerase-beta.

Hyperthermia has been shown to inhibit the activity of DNA polymerase-beta when either the intact Chinese hamster cells or partially purified enzyme samples from CHO cells are heated (42.2 or 45.5 degrees C). However, the loss of activity from heating isolated enzyme samples can be either greater or less than the loss from heating intact cells. For example, treating cells with the membrane-active agent procaine-HCl greatly sensitizes the cells to heat-induced loss of enzyme activity but has no effect on the heat sensitivity of isolated enzyme. Furthermore, the heat sensitivity of the isolated enzyme depends greatly on the purification steps and can be reduced by heating the enzyme in the presence of bovine serum albumin, activated DNA, or Langendorf salts. These observations, considered in relation to others in the literature, suggest that heat inactivation of polymerase-beta occurs from heat-induced changes in the intracellular environment, which in turn modify the direct thermal denaturation of the enzyme within the cell.

Animals↗
PubMed1982

The effect of chronic and acute heat conditioning on the development of thermal tolerance.

Survival studies with Chinese hamster ovary cells showed that thermal tolerance, which developed during chronic heating (treatment times greater than or equal to 1 hr) or after acute heating (treatment times less than 1 hr) involves similar mechanisms. For example, cells that expressed thermal tolerance during a 6-14 hr chronic heat treatment at 41.5 degrees C or 42 degrees C also expressed thermal tolerance to a subsequent acute treatment at 45.5 degrees C. Also, cells heated acutely for 10 min at 45.5 degrees C and incubated at 37 degrees C for 12 hr showed tolerance to both 45.5 degrees C acute and 42 degrees C chronic hyperthermia. Finally, thermal tolerance developed between fractionated acute heat treatment at 45.5 degrees C and fractionated chronic heat treatments at 42.5 degrees C. These data indicate that when cells are tolerant to chronic hyperthermia they are also tolerant to acute hyperthermia and that the reverse is also true.

Adaptation, Physiological↗
PubMedDate not supplied

Measurement of dose distribution around Fletcher-Suit-Delcos colpostats using a Therados radiation field analyzer (RFA-3).

Using a small silicon diode and the Therados RFA-3 automated dosimetry system, the relative dose distribution around Fletcher-Suit-Delcos colpostats was measured. Thermoluminescent dosimeters (LiF) were used for absorbed dose measurements. These data provide the basis for computerized dose calculations which will account for the attenuation effects of the rectal and bladder shieldings. The stainless-steel housing attenuated the radiation of a Cs-137 source by 6%. The tungsten shields caused a further dose reduction of 15%-25%.

Brachytherapy↗
PubMedDate not supplied

Normal cells and malignant cells transfected with the HRas oncogene have the same heat sensitivity in culture.

Mouse embryo cells (C3H 10T1/2) were transfected with a plasmid (pAL8A) containing the HRas oncogene and neomycin resistance gene. Five transfected cell clones were isolated and established as cell lines, and these showed neomycin resistance. Two of these cell lines expressed a normal morphology while three showed a transformed morphology. Four of the cell lines produced tumours in nude mice. Flow cytometry measurements showed that exponentially growing cell cultures of the five transfected cell lines had the same cell cycle distribution as the normal parental cell line. The sensitivity to hyperthermia of the five transfected cell lines was the same as that of the normal cell line for temperatures ranging from 42.0 to 45.0 degrees C. Thus in these studies we found no difference in the thermal sensitivity of normal and malignant cells transfected by the Hras oncogene.

Cell Line↗
PubMedDate not supplied

Role of accelerated fractionated irradiation for supraglottic carcinoma: assessment of results.

PURPOSE: We evaluated the results of locoregional control, patients' relapse-free survival, and voice preservation in patients with supraglottic carcinoma treated with accelerated radiation therapy. PATIENTS AND METHODS: The records of 164 patients undergoing accelerated fractionated radiation therapy for carcinoma of the supraglottis from 1981 to 1992 were reviewed and evaluated for locoregional control, disease-specific survival, and rates of voice preservation. All patients were treated with 1.6 Gy per fraction two fractions a day (BID) for 67.2 to 70.0 Gy in 6 weeks. The median follow-up was 56 months. Due to severe acute mucosal toxicity, all patients were given a short treatment break after 38.4 to 48.0 Gy. RESULTS: For T1, T2, T3, and T4 tumors, the 5-year actuarial local control rates were 96%, 86%, 76%, and 43%, respectively, and relapse-free survival rates were 78%, 82%, 64%, and 40%, respectively. With surgical salvage, the corresponding ultimate local control rates for the T1, T2, T3, and T4 tumors were 96%, 93%, 88%, and 51%, respectively. Regional control by T and N stage was non-significant. Voice preservation rates for the T1, T2, T3, and T4 tumors were 96%, 80%, 72%, and 43%, respectively, and for the entire group the rate was 79%. CONCLUSIONS: Accelerated fractionated radiation therapy resulted in excellent locoregional control, relapse-free survival, and voice preservation rates for patients with supraglottic carcinoma. The T stage and N stage were significant predictors of outcome. T4 tumors and node-positive neck disease portended a poor prognosis. These results as compared to our historical control and the published data in the literature with conventional once-daily radiation therapy (QD) suggested improved local control and patient survival after accelerated fractionated BID radiation therapy. For confirmation of these results, a prospective randomized trial is needed.

Carcinoma, Squamous Cell↗