A cabbage looper's job, in many respects not unlike our own, is to survive its early developmental period and grow up healthy, metamorphose into an adult moth, and beget the next generation of loopers. Given its numerous predators, exposure to the continuous barrage of toxic physical, chemical and biological agents delivered by humans in an effort to eradicate it, as well as nature's own hazards, such as the ultraviolet component of sunlight and hazardous natural chemicals in plants that serve to deter ingestion by the looper, it is not difficult to view the looper as a highly stressed organism. However, the looper and its lepidopteran ancestors have been quite successful at resisting such hazards since the Mesozoic Era. The purpose of this brief review will be to examine some of the potential mechanisms by which the looper and its relatives have been able to be so successful over these past 200 million years.

Tissue expansion facilitates coverage of cutaneous defects both through the generation of additional skin surface area and by increasing random flap length/width viability factors. The opportunity to apply tissue expansion techniques to head and neck defects within previously irradiated skin fields continues to increase. To study the effect of tissue expansion on the cutaneous perfusion and wound-healing capacity of irradiation-damaged skin, rabbit scalps were subjected to 5 weeks of fractionated radiation followed at 4 months by prolonged tissue expansion. Standardized random flaps were then created and reset within the expanded skin and analyzed in parallel with nonirradiated and nonexpanded control animals. Flap viability as expressed by area and mean maximum length was determined at 10 days postwounding followed by determination of wound-breaking strength. Irradiated tissues demonstrated a significantly reduced flap viability that was significantly increased by expansion. However, tissue expansion-related increases in flap length exceeded those expressed as percent of total area surviving for irradiated animals. Tissue expansion resulted in significant increases in wound tensile strength only in nonirradiated animals. These findings suggest that, compared with controls, several of the benefits of tissue expansion are less appreciable in radiation-damaged skin.

Opinion remains divided over the advisability of tissue expansion in previously irradiated skin. We examined the properties of, and complications associated with, tissue expansion in previously irradiated rabbit scalps. Irradiation injury was produced using fractionated roentgen rays, with a total dose of 5000 cGy over a 5-week interval. Following a 20-week convalescence interval, expansion was incrementally conducted over 4 weeks. Monitored parameters included cutaneous perfusion as indicated by fiberoptic dermofluorometry, intraluminal pressure, linear surface gain, and area of surface necrosis. The incidence and severity of complications, including surface necrosis, were significantly higher among irradiated animals. Furthermore, the overlying skin of irradiated animals demonstrated a significantly decreased compliance and measurable area gain. Given the inferior expansibility and higher tendency toward complications with contemporary expansion techniques in previously irradiated skin, alternate reconstructive options are preferable in this setting.

Human coronary artery restenosis after percutaneous revascularization is a response to mechanical injury. Smooth muscle cell proliferation is a major component of restenosis, resulting in obstructive neointimal hyperplasia. Because ionizing radiation inhibits cellular proliferation, this study tested in a porcine coronary injury model the hypothesis that the hyperplastic response to coronary artery injury would be attenuated by X-irradiation.Deep arterial injury was produced in 37 porcine left anterior descending coronary artery segments with overexpanded, percutaneously delivered tantalum wire coils. Three groups of pigs were irradiated with 300-kV X-rays after coil injury: Group I (n = 10), 400 cGy at 1 day; Group II (n = 10), 400 cGy at 1 day and 400 cGy at 4 days and Group III (n = 9), 800 cGy at 1 day. Eight pigs in the control group underwent identical injury but received no radiation. Treatment efficacy was histologically assessed by measuring neointimal thickness and percent area stenosis.Mean neointimal thickness in all irradiated groups was significantly higher than in the control groups and thickness was proportional to X-ray dose.X-irradiation delivered at these doses and times did not inhibit proliferative neointima. Rather, it accentuated the neointimal response to acute arterial injury and may have potentiated that injury.

Levamisole in combination with radiation and chemotherapeutic agents is being studied in clinical trials. The mechanism of interaction of levamisole with these modalities is unknown. In order to determine if there is direct interaction between radiation and levamisole, a series of colony-formation assays was performed with the use of two human tumor cell lines. Cells were exposed to 0-10 Gy of radiation, with or without the addition of 0-1000 microM levamisole. Exposure to levamisole alone had no effect on cell survival; however, the combination of continuous exposure to levamisole at concentrations approaching 1000 microM and radiation revealed a potentiation of radiation-induced cell killing.

Two gamma-ray-sensitive and two ultraviolet (UV)-sensitive variants were isolated from the gamma-ray- and UV-resistant TN-368 lepidopteran insect cell line. The isolation was performed by inducing mutations in the TN-368 cells using ethyl methanesulfonate, growing them for an expression period, irradiating with 137Cs gamma rays or 254-nm UV radiation, allowing cells to incorporate 5-bromodeoxyuridine (BrdU) in the presence of hydroxyurea (DNA repair synthesis), and finally irradiating with 365-nm UV radiation to cause DNA strand breakage at sites of BrdU incorporation with the intent of killing those cells that have undergone DNA repair synthesis and sparing those cells which, for a variety of reasons, did not. The survival of the Cs2 and Cs7 variants exposed to X rays is significantly different from the parent TN-368 line at the P less than 0.0001 level. The survival of the UV10 and UV19 variants exposed to UV radiation is different from the parent at the P less than 0.0001 and P less than 0.003 levels, respectively. In cross-sensitivity testing of the gamma-ray-sensitive variants, only Cs2 is more sensitive to 254-nm UV and only Cs7 is more sensitive to 44 degrees C heating; both are sensitive to PUVA. The UV-sensitive mutants are both sensitive to X irradiation, PUVA, and mitomycin C. However, UV10 is not sensitive to 44 degrees C heating while UV19 is, making UV19 the only variant strain sensitive to all agents examined. Despite the isolation procedure which was intended to select for DNA repair-deficient cells, the results suggest that a more general mechanism is responsible for the sensitivity of the variant cells to the agents tested.

TN-368 lepidopteran insect cells are on the order of 100 times more resistant to the lethal effects of ionizing radiation than cultured mammalian cells. DNA double-strand breaks (DSB) are believed by many to be the critical molecular lesion leading to cell death. We have therefore compared the rejoining of DSB in TN-368 and V79 Chinese hamster cells. Cells were irradiated on ice with 137Cs gamma rays at a dose rate of 2.5 Gy/min, incubated for various periods of time, and assayed for DNA DSB using the method of neutral elution. The kinetics of DSB rejoining following a dose of 90.2 Gy is similar for both cell lines with 50% of the rejoining completed in about 12 min. Approximately 83 and 87% of the DSB are rejoined in the TN-368 and V79 cells, respectively, by 1 h postirradiation. However, no further rejoining occurs in the TN-368 cells through at least 6 h postirradiation, whereas approximately 92% of the DSB are rejoined in the V79 cells by 2 h postirradiation. Other studies (from 22.6 to 226 Gy) demonstrate that the amount of rejoining of DSB varies inversely with dose for both cell lines, but this relationship is not as pronounced for the TN-368 cells. In general, these findings do not support the hypothesis that unrejoined DNA DSB represent the critical molecular lesion responsible for cell death.

TN-368 lepidopteran insect cells display a pronounced resistance to the lethal effects of ionizing radiation and exhibit superior DNA repair capabilities. When a TN-368 cell population entering stationary growth phase is irradiated with 137Cs gamma rays and then incubated for several hours before cell dilution and plating for colony formation, the surviving fraction is increased several-fold over cells diluted and plated immediately after irradiation. Similarly, the survival of cells plated immediately following the second of two equivalent doses separated by several hours is greater than the survival of cells plated immediately following a single dose equal to the sum of the split doses. Both processes exhibit similar biphasic repair kinetics and reach maximal levels by 6 h. The phenomena appear initially to be analogous to confluent-holding and split-dose recovery as described for mammalian cells. However, the survival levels obtained for doses of 61-306 Gy after allowing for these recovery processes to occur are quite high and greatly exceed survival levels for all but relatively low doses less than 50 Gy. For example, while the survival of cells irradiated with 150 Gy is near 0.15, the survival of cells receiving 306 Gy in two equivalent split doses is approximately 0.77. Even if damage induced by the first of the split doses was completely repaired, it might be expected that the survival would be near the level of the second dose alone, or near 0.15. Instead the survival is approximately five times greater, suggesting that the first split dose stimulated a repair system not present in unirradiated cells. The situation for confluent-holding recovery is similar to that for split-dose recovery.

The filter elution technique was used to assay for 137Cs-induced DNA double-strand breaks (dsb) in V79 Chinese hamster cells. The elutions were performed using two different sets of lysing and eluting solutions at pH 7.2 and 9.6. The data agree with those of others who have demonstrated differences in elution profiles at pH 7.2 and 9.6. In addition, the data indicate that solution composition has a significant effect on elution.

Cell survival and photoreactivation of 254 nm ultraviolet (UV) light damage in a wild typeDrosophila cell line was assayed by colony formation in liquid medium. Fo, Fq, and extrapolation number for the exponential portion of survival curves are 21 J/m2, 3.6 J/m2, and 1.5 for non-photoreactivated cells and 110 J/m2, 11.2 J/m2, and 1.3 for those exposed to photoreactivating light. Maximal photoreactivation occurs at the 100 J/m2 region of the curve. At 10 and 50% survival, 75–80% of the UV damage was photoreactivable.

Purpose: To retain cell viability, mammalian cells can increase damage repair in response to excessive radiation-induced injury. The adaptive response to small radiation doses is an example of this induced resistance and has been studied for many years, particularly in human lymphocytes. This review focuses on another manifestation of actively increased resistance that is of potential interest for developing improved radiotherapy, specifically the phenomenon in which cells die from excessive sensitivity to small single doses of ionizing radiation but remain more resistant (per unit dose) to larger single doses. In this paper, we propose possible mechanisms to explain this phenomenon based on our data accumulated over the last decade and a review of the literature.Conclusion: Typically, most cell lines exhibit hyper-radiosensitivity (HRS) to very low radiation doses (<10 cGy) that is not predicted by back-extrapolating the cell survival response from higher doses. As the dose is increased above about 30 cGy, there is increased radioresistance (IRR) until at doses beyond about 1 Gy, radioresistance is maximal, and the cell survival follows the usual downward-bending curve with increasing dose. The precise operational and activational mechanism of the process is still unclear, but we propose two hypotheses. The greater amount of injury produced by larger doses either (1) is above a putative damage-sensing threshold for triggering faster or more efficient DNA repair or (2) causes changes in DNA structure or organization that facilitates constitutive repair. In both scenarios, this enhanced repair ability is decreased again on a similar time scale to the rate of removal of DNA damage.

Early alterations in normal semiconservative and repair DNA synthesis were determined in gastrointestinal tissues of HalCR mice following administration of the colon carcinogen 1,2-dimethylhydrazine (DMH). Following DMH injections of 60 and 200 mg/kg, normal DNA synthesis was rapidly inhibited in all tissues. The greatest depressions were observed in the descending colon, followed closely by the ascending colon. DNA repair was estimated by measuring unscheduled DNA synthesis. No repair was observable in the descending or ascending colon. The esophagus, forestomach, jejunum, and ileum demonstrated significant amounts of DNA repair, while the duodenum and gastric stomach displayed nominal or insignificant amounts of repair. Repair DNA synthesis was inhibited by simultaneous administration of caffeine and DMH. The degree of inhibition of normal replicative DNA synthesis and the amount of repair DNA synthesis in response to DMH treatment correlate closely with the incidence of DMH-induced tumors. Most tumors occur in the descending colon, followed by the ascending colon, and only a few in the duodenum and gastric stomach area. Neoplasms are rarely found in the remainder of the gastrointestinal system.

TN-368 lepidopteran insect cells display a multiphasic survival response in both air and nitrogen. In each case the survival curve is characterized by an initial small- shouldered component having a steep slope, a plateau or broad- shouldered region near the 0.1 survival level, and finally a shallow slope component. The D0, Dq, and n values for the initial steep slope component in air and nitrogen are, respectively, 65.7 Gy, 9.0 Gy, and 1.2, and 104.4 Gy, 28.8 Gy, and 1.3. The oxygen enhancement ratio (OER) for this portion of the curve is 1.6. The D0, Dq, and n values for the shallow slope component in air and nitrogen are, respectively, 130.2 Gy,--36.1 Gy, and 0.8, and 226.8 Gy, 121.0 Gy, and 1.7. The OER for this portion of the curve is 1.7. The D0 values for each slope and the width of the plateau region all increase proportionally for the nitrogen curve over that of air, the OER being approximately the same for both curve components. A similar multiphasic response was observed at dose rates of 202, 49.6, and 9.1 Gy/min. In addition, the survival of cells which had previously been irradiated with a dose well into the logarithmic region of the more resistant shallow slope portion of the curve retained a multiphasic response. Although cell cycle variations in radiosensitivity may contribute slightly to the response, an inducible or activated repair process would be consistent with the results.

The radiosensitivity of five dipteran cell lines representing three mosquito genera and one fruit fly genus were examined. These lines are: (1) ATC-10, Aedes aegypti; (2) RU-TAE-14, Toxorhynchites amboinensis; (3) RU-ASE-2A, Anopheles stephensi; (4) WR69-DM-1, Drosophila melanogaster; and (5) WR69-DM-2, Drosophila melanogaster. Population doubling times for these lines range from approximately 16 to 48 hr. Diploid chromosome numbers are six for the mosquito cells and eight for the fruit fly cells D/sub 0/ values are 5.1 and 6.5 Gy for the Drosophila cell lines and 3.6, 6.2, and 10.2 Gy for the mosquito cell lines. The results of this study demonstrate that dipteran insect cells are a few times more resistant to radiation than mammalian cells, but not nearly as radioresistant as lepidopteran cells.

Abstract— Unscheduled DNA synthesis has been measured at several times during the differentiation of cultured rat skeletal muscle cells in response to exposures to 254nm UV light. There is no change in the amount of repair DNA synthesis as the cells fuse and differentiate from postmitotic prefusion myoblasts to multinucleated contracting myotubes.