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Single dose or fractionated total body irradiation and autologous marrow transplantation in dogs: effects of exposure rate, fraction size, and fractionation interval on acute and delayed toxicity.

Dogs were given single dose or fractionated total body irradiation (TBI) and autologous marrow grafts to prevent death from myelosuppression. Acute and delayed non-marrow toxicities were compared. Fifty-six dogs were given single dose TBI at 2.1 (n = 13), 5 (n = 12), 10 (n = 15), or 20 (n = 16) cGy/min. Acute radiation toxicity and mortality was related to the exposure rate; radiation doses resulting in 50% mortality at 7 days (LD 50/7) at 2.1, 5, 10 and 20 cGy/min were 1,692, 1,499, 1,261, and 1,056 cGy respectively. Fifty-three dogs were given fractionated TBI, 200 cGy three times a day with 6-hour intervals at 2.1 (n = 13), 5 (n = 9), 10 (n = 13), or 20 (n = 18) cGy/min. The LD 50/7 at the four exposure rates were 1,628, 1,470, 1,184, and 1,320 respectively. Thus, for exposure rates of 2.1, 5, and 10 cGy/min, the tolerated doses were comparable for single dose and fractionated TBI. At 20 cGy/min dose fractionation appeared to offer some advantage, although this fractionation effect in part may have been due to random variation with small numbers of dog treated. Following recovery from the immediate TBI-related toxicity, eight dogs given single dose and four dogs given fractionated TBI died, generally from infections, 8-30 days following transplantation. There was a striking difference in regards to long-term survival dependent upon the TBI regimen. Among dogs given greater than or equal to 1,000 cGy of TBI and alive 30 days after transplant only 1 of 18 given single dose TBI became a long-term survivor compared to 19 of 22 given fractionated TBI. Causes of death included pancreatic fibrosis, malnutrition, hepatic failure, and a generalized wasting syndrome. All 5 dogs given a single dose of 800 cGy (at 20 cGy/min) became long-term survivors. Fourteen dogs were given increments of 150 cGy at 7 cGy/min every 3 hours for total doses of 1,500-2,400 cGy. The LD 50/7 was approximately 1,900 cGy. All 6 dogs alive at 30 days became healthy long-term survivors. Four dogs were given increments of 600 cGy at 2.1 cGy/min every 48 hours for a total dose of 1,800 cGy. All 4 dogs became long-term survivors. In conclusion, exposure rate and total dose are the most important parameters for acute toxicity associated with TBI. The effect of dose fractionation is minimal at low exposure rates and appears to be dependent also upon increment size and fractionation interval.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

The application of the linear-quadratic model to fractionated radiotherapy when there is incomplete normal tissue recovery between fractions, and possible implications for treatments involving multiple fractions per day.

By extending a previously developed mathematical model based on the linear-quadratic dose-effect relationship, it is possible to examine the consequences of performing fractionated treatments for which there is insufficient time between fractions to allow complete damage repair. Equations are derived which give the relative effectiveness of such treatments in terms of tissue-repair constants (mu values) and alpha/beta ratios, and these are then applied to some examples of treatments involving multiple fractions per day. The interplay of the various mechanisms involved (including repopulation effects) and their possible influence on treatments involving closely spaced fractions are examined. If current indications of the differences in recovery rates between early- and late-reacting normal tissues are representative, then it is shown that such differences may limit the clinical potential of accelerated fractionation regimes, where several fractions per day are given in a relatively short overall time.

Cell Survival

A rapid Percoll gradient procedure for isolation of synaptosomes directly from an S1 fraction: homogeneity and morphology of subcellular fractions.

A method for preparation of synaptosomes from rat cerebral cortex, on a discontinuous Percoll gradient, was previously developed for use with a P2 pellet (Brain Research, 372 (1986) 115-129). Here the Percoll method has been adapted for use with an S1-supernatant which eliminates a potentially damaging resuspension step and saves over 30 min, representing a third of the total preparation time. The homogeneity of the synaptosomes in each of the 5 subcellular fractions obtained with the S1-Percoll method was determined biochemically by analysis of the distribution of total protein, myelin basic protein, synapsin I and pyruvate dehydrogenase across the gradient. Electron microscopy was also used to determine the homogeneity of the synaptosomes, as well as to determine their morphological characteristics. Fraction 4 was the most enriched in synaptosomes and contained the lowest level of contamination by myelin, extrasynaptosomal mitochondria and plasma membranes. The yield of synaptosomes in fraction 4 with the S1-Percoll method was 1.4-fold greater than with the P2-Percoll method. While all other fractions contained some synaptosomes the major additional content in fractions 1-3 and 5 was, respectively, unidentified small membranes, myelin, synaptic plasma membranes and extrasynaptosomal mitochondria. Fraction 1 was enriched for very small synaptosomes (0.34 micron mean diameter) only 8% of which contained mitochondria, while fractions 2-4 progressively included larger synaptosomes containing more mitochondria. Fraction 5 synaptosomes were approximately the same size as those in fraction 4 (0.63 micron mean diameter), but 83% contained mitochondria, significantly more than in fraction 4. The synaptosomes in fraction 5 were found to be relatively resistant to hypotonic lysis, explaining a previously observed lack of phosphorylation of synapsin I in this fraction. The differences in homogeneity and morphological characteristics of the synaptosomes in fractions 1-5 suggest that the basis for their fractionation on Percoll gradients is different from that achieved with the more traditional procedures for isolating synaptosomes and that unique synaptosomal fractions are obtained with the S1-Percoll procedure.

Animals

Studies on proinsulin and proglucagon biosynthesis and conversion at the subcellular level. I. Fractionation procedure and characterization of the subcellular fractions.

Anglerfish islets were homogenized in 0.25 M sucrose and separated into seven separate subcellular fractions by differential and discontinuous density gradient centrifugation. The objective was to isolate microsomes and secretory granules in a highly purified state. The fractions were characterized by electron microscopy and chemical analyses. Each fraction was assayed for its content of protein, RNA, DNA, immunoreactive insulin (IRI), and immunoreactive glucagon (IRG). Ultrastructural examination showed that two of the seven subcellular fractions contain primarily mitochondria, and that two others consist almost exclusively of secretory granules. A fifth fraction contains rough and smooth microsomal vesicles. The remaining two fractions are the cell supernate and the nuclei and cell debris. The content of DNA and RNA in all fractions is consistent with the observed ultrastructure. More than 82 percent of the total cellular IRI and 89(percent) of the total cellular IRG are found in the fractions of secretory granules. The combined fractions of secretory granules and microsomes consistently yield >93 percent of the total IRG. These results indicate that the fractionation procedure employed yields fractions of microsomes and secretory granules that contain nearly all the immunoassayable insulin and glucagons found in whole islet tissue. These fractions are thus considered suitable for study of proinsulin and proglucagon biosynthesis and their metabolic conversion at the subcellular level.

Animals

Accelerated versus conventional fractionation. The degree of incomplete repair in human skin with a four-hour-fraction interval studied after postmastectomy irradiation.

A previously presented clinical assay with postoperative irradiation to bilateral parasternal fields in patients with breast cancer was used for a comparison of acute and late reactions in human skin after accelerated and conventional fractionation. Two and 3 fractions per day at 4-hour intervals were compared with one fraction per day. Dose fractions of about 2 Gy were used. Twenty-five fractions were given in 2.5, 1.5 and 5 weeks respectively. The acute reactions were similar regardless if 1, 2 or 3 fractions per day were given, i.e. equal total doses were isoeffective. The repair of intracellular damage was apparently completed within 4 hours. However, this might not be true due to a differential influence of proliferation and redistribution on the effect of different types of fractionation, which makes it difficult to interpret the result and estimate the degree of intracellular repair. The time to the acute peak reaction was shortened by 6 to 7 days with accelerated compared to conventional fractionation, explained by the differences in the dose delivery rates. Consequently, the onset of a compensatory proliferation is earlier after accelerated fractionation. Late reactions were more pronounced after accelerated than after conventional fractionation and 1 X 2.0 Gy/day was found to be equivalent to 2 X 1.80 Gy/day and 3 X 1.65 Gy/day at 4-hour intervals with an equal fraction number for all 3 schedules. Assuming that proliferation is negligible for late responding tissues, we interpret this finding as an expression of the degree of reduced intracellular repair. Finally, we would like to point out that the iso-effect dose relationships between acute and late reactions for accelerated versus conventional fractionation might vary, above all with the cell proliferation kinetics of acutely reacting tissue.

Breast Neoplasms

[Concentration of components of the adenylate system in heavy and light fractions of the sarcoplasmic reticulum of skeletal muscles and sensitivity of these fractions to the effects of imidazole-containing compounds and caffeine].

Fragments of sarcoplasmic reticulum from rabbit sceletal muscles sedimented within the range from 2000 g to 8000 g (heavy fraction) and 8000 g to 40000 g (light fraction) and washed with 0.6 M KCl, were practically free of adenylatecyclase activity. Phosphodiesterase cAMP was not found in the light fraction, while its activity in the heavy fraction was 500 pmol of cAMP/min per mg of protein. Both fractions contain bound cAMP (1-2 pmol/mg of protein) and specific sites of cAMP binding, the binding constant being approximately 10(6)M-1. The number of binding sites is 60 pmol/mg of protein for the heavy and 30 pmol/mg of protein for the light fractions. The level of phosphodiesterase activity in the heavy fraction correlates with its sensitivity to imidazole, anserine and caffeine. Imidazole and anserine increase in 1.5-1.8 times the value of Ca2+/ATP in the heavy fraction and produce no effect on Ca2+ transport by the light fraction. Caffeine decreases almost twice the Ca2+/ATP value in the heavy fraction and has practically no effect on Ca2+ absorption by enzymes of the light reticulum fraction. Imidazole and anserine activate membrane-bound phosphodiesterase, while caffeine inhibits it. It is suggested that structural rearrangements of membrane-bound phosphodiesterase under the effect of caffeine, imidazole and anserine are responsible for changes in the efficiency of Ca2+ transport by fragments of the heavy reticulum fractions.

Adenosine Triphosphatases

A rapid Percoll gradient procedure for isolation of synaptosomes directly from an S1 fraction: viability of subcellular fractions.

The metabolic and functional viability of synaptosomes was examined in 5 subcellular fractions obtained after centrifugation of an S1 fraction from rat cerebral cortex on a discontinuous Percoll gradient (Brain Research, this volume, 1987). Fraction 4 was the most enriched for viable synaptosomes since, although it accounted for only 11.8% of the total protein recovered from the gradient, this fraction contained 23.7% of the basal synapsin I phosphorylation activity, the greatest degree of depolarisation-stimulated increase in synapsin I phosphorylation, 36.1% of the total [3H]noradrenaline uptake capacity and 46.9% of the total [3H]noradrenaline release capacity. Noradrenaline release from fraction 4 was consistent with a neuronal mechanism as it was increased with increasing K+ concentrations and was dependent on calcium. Fractions 1 and 2 contained few viable synaptosomes as judged by their capacity for noradrenaline uptake and release, yet these fractions accounted for some 62.6% of the endogenous content of noradrenaline. In part their lack of viability was due to a low content of intrasynaptosomal mitochondria, while their high content of endogenous noradrenaline was due to the presence of synaptic vesicles released from damaged nerve terminals. The synaptosomes in fraction 3 were metabolically and functionally viable, but their capacity for uptake and release of noradrenaline was lower than for fraction 4. The synaptosomes in fraction 5 showed only a small depolarisation-stimulated release of noradrenaline, suggesting a lack of viability. Part of the capacity for uptake of [3H]noradrenaline into fraction 5 was attributed to the presence of extrasynaptosomal mitochondria.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Specific-locus mutation response to unequal, 1 + 9 Gy X-ray fractionations at 24-h and 4-day fraction intervals.

The specific-locus mutation frequency obtained from mouse spermatogonial stem cells following unequal, 1 + 9 Gy X-ray fractionation with a 24-h fractionation interval is low, and consistent with the two fractions acting additively. The response is therefore markedly different from the augmented mutation frequencies obtained with 500 + 500 R and 100 + 500 R, 24-h fractionations. The lower yield compared with the 100 + 500 R response also indicates a clear difference from the translocation data which demonstrate increases in yield with increasing second dose over the same dose range. The decline in specific locus mutation yield with the increase in the second dose from 500 R to 9 Gy suggests that the stem cells surviving the first fraction are heterogeneous in their sensitivities to this class of genetic damage. A similar, additive specific locus mutation frequency is obtained with unequal, 1 + 9 Gy X-irradiation when the interval between fractions is 4 days. This is consistent with 500 + 500 R, 4-day and 7-day interval responses obtained previously but again differs from the sub-additive translocation responses obtained with such X-ray fractionation. Taken together with the data from previous studies the present results suggest that (1) 24 h after the first fraction, (a) the surviving stem cell have two components; survivors of the formerly radiosensitive, cycling component of the normal stem cell population and the formerly radioresistant, G0 or arrested G1 cells, which are being 'triggered' into a rapid cell cycle to achieve repopulation of the testis; (b) these two components are of near-equal sensitivity to translocation induction and cell killing, hence the additive translocation yields with equal X-ray fractionations and yields consistent with those extrapolated from lower doses with higher, unequal fractionations, e.g. 1 + 7 Gy, 1 + 9 Gy; but (c) the formerly radioresistant, triggered component is much more sensitive than the surviving cycling component to specific locus mutation and cell killing, hence the augmented mutation response with 500 + 500 R fractionation and the drop in yield with 1 + 9 Gy compared with 100 + 500 R X-irradiation.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Subcellular fractionation of a hypercellulolytic mutant, Trichoderma reesei Rut-C30: localization of endoglucanase in microsomal fraction.

The growing mycelia of Trichoderma reesei Rut-C30 are richly endowed with endoplasmic reticula and a variety of pleomorphic subcellular bodies. Mycelia of the culture growing in presence of avicel pH101 was fractionated in sucrose density gradients, and several morphologically and biochemically distinct fractions were isolated. Mycelia were homogenized in a Bead Beater, and the homogenate was freed of nucleus and wall fragments by low-speed centrifugation before fractionation. Organelle-free cytosol, which did not penetrate the gradient, contained (of the total) 72% of the vanadate-sensitive ATPase, 26% of carboxymethyl cellulase (CMCase), 2% of cytochrome c reductase, and 13% of the protein. Significant fractions separated on a gradient were light vesicles containing heavily stained material inside and ribosomes attached to the outside surface, intact vesicles resembling condensing vacuoles, large vesicles derived from the plasma membrane, and heavy vesicles containing crystalline material. The light-vesicle fraction contained a large portion of the cell-bound CMCase activity. The particle-bound ATPase and cytochrome c reductase activities were concentrated in heavy fractions. The fractionation in the presence of MgCl2 improved the preservation of subcellular bodies derived from the endoplasmic reticula. Although the CMCase activity of the light-vesicle fraction was 4 times higher than the activity in the heavy-vesicle fraction, the CMCase antibody-binding capacities of both fractions were about the same. This discrepancy between the catalytic activity and the antibody-binding capacity suggests that the heavy vesicles might have contained considerable amount of inactive CMCase compared with that present in the light vesicles.

Cell Fractionation

Fractionation of the proteolytic and amylolytic complex enzyme system of streptomyces aureofaciens and some properties of fractions.

The Streptomyces aureofaciens extracellular proteolytic system was split into four fractions by carboxymethylcellulose (CMC) column chromatography giving three purely caseinolytic fractions and one fraction active toward both starch and casein. The first caseinolytic and amylolytic fraction was further fractionated by DEAE-Sephadex A-50 chromatography into one purely amylolytic fraction and another showing both activities, was refractioned into four new fractions by DEAE-cellulose chromatography. These fractions were found to be heterogeneous by polyacrylamide gel electrophoresis, three of them acted on both starch and casein and a fourth was only caseinolytic. The second CMC fraction was further purified by CMC rechromatography to an homogeneous fraction that hydrolyzes carboxypeptidase A(EC 3.4.2.1) synthetic substrates and solubilizes elastin. It had only one polypeptide chain with a molecular weight of about 28000 daltons, a high thermal stability in the presence of calcium ions, a pH optimum of about 6.8, and a maximal caseinolytic activity at about 50 degrees C.

Amylose

The 10-year results of a prospective trial of post-operative radiotherapy delivered in 3 fractions per week versus 2 fractions per week in breast carcinoma.

The 10-year results are presented of a prospective trial of 411 patients with breast carcinoma treated by mastectomy and post-operative radiotherapy given in either 2 or 3 fractions per week (i.e. a comparison of 6 fractions in 18 days with 12 fractions in 28 days). The early radiation effects on the normal tissues were similar and acceptable. The late skin changes in the chest wall (treated with 70 kV X rays) were progressive and by 10 years were slightly more marked with 6 fractions. Late subcutaneous fibrosis in the axilla (treated with cobalt-60 teletherapy), however, was much less in the 6-fraction group. Twelve fractions resulted in greater restriction of shoulder movement and an increased incidence of lymphoedema of the arm. Doses were selected on the basis of past clinical experience. The dose used to treat the axilla in 6 fractions was 35 Gy, 14.99% less than that predicted by NSD. The dose predicted by alpha:beta, using a value of 2 Gy for late reactions, is 38.14 Gy. Thus simple theory, which omits time, still predicts too high a dose for 6 fractions, although it is closer than NSD. In this trial, the 6-fraction technique showed an advantage over the 12-fraction technique. It was equally effective in controlling local recurrence and had fewer late sequelae. It was also convenient for patients and economic in the use of radiotherapy resources.

Breast Neoplasms

Thermotolerance induced by fractionated hyperthermia: dependence of the interval between fractions.

The induction of thermotolerance by fractionated hyperthermia was investigated in the mouse ear. Ears were heated at 43.5 degrees C by immersion in water. One to ten treatments of 20 min were followed by test treatments. Thermotolerance was assessed as the increase in the duration of the test treatment required for a thermal response in 50 per cent of the ears (NT50). A single treatment induced thermotolerance which reached a maximum at 24 h when the NT50 was increased by a factor of 2.4. The same maximum was observed after each fractionated treatment used in the present study. The time course of development, however, depended on the interval between fractions. (1) When the interval was too short to allow development of thermotolerance after a single fraction (4 h), thermotolerance was not induced during fractionated treatment but it developed during the first 24 h after treatment. (2) When the interval between fractions allowed the maximal development of thermotolerance (24 h), this maximum was maintained during fractionated treatment and persisted for 24 h after treatment. (3) When the interval allowed some decay of thermotolerance (72 or 168 h) there was a further increase to maximal thermotolerance after each fraction. The decay of thermotolerance from the maximum did not depend on the interval between fractions. These results indicate that the degree of thermotolerance may fluctuate during fractionated hyperthermia.

Adaptation, Physiological

Large-scale fractionation of S-form lipopolysaccharide from Salmonella abortus equi. Chemical and serological characterization of the fractions.

The S-form lipopolysaccharide of Salmonella abortus equi was separated by a newly elaborated extraction method with organic solvents into three fractions of different chain length of the O-polysaccharide they contained. The three fractions were designated long-chain (20-50 repeating units), short-chain (0-6) and R-fraction (no repeating units) according to their migration pattern in polyacrylamide gel electrophoresis in the presence of sodium dodecylsulphate. The nature of the fractions as long- and short-chain and as R-fraction was confirmed by chemical analysis. The concentration of O-specific sugars was highest in the long-chain fraction, where their molar ratio to glucosamine was ca. 25:1. In the short-chain fraction the ratio of O-sugars to glucosamine was 2.5:1, and in the R-fraction O-specific sugars were absent. The serological properties of the three fractions were in good agreement with their chemical composition.

Carbohydrates

Antigens of the subcellular fractions of Trypanosoma cruzi. II. Flagellar and membrane fraction.

A method for isolation of membrane and flagellar fractions from Trypanosoma cruzi epimastigotes, and descriptions of their ultrastructural characteristics and antigenic activity are presented. Electron-microscopic observation revealed that the membrane fraction resembled vesicles, as occurs frequently when the pressure-depressure method is used for cell disruption. The flagellar fraction revealed flagella and some membranes, both related and unrelated to the flagellar structure. Flagellar and membrane fractions had 5 and 3 precipitin lines respectively against anti-whole homogenate of T. cruzi, in double diffusion tests. One of the precipitin bands common to both fractions gave a reaction of identiy with one precipitin line of the microsomal fractions, that was not present in the mitochondrial, nuclear and cell sap fractions. Data from tests of protective activity activity obtained during a period of 3 years with whole homogenate and F and M fraction are also presented Protective activity against lethal challenge doses of trypomastigotes is strongly associated with the flagellar fraction.

Animals

Two fractions of rough endoplasmic reticulum from rat liver. II. Cytoplasmic messenger RNA's which code for albumin and mitochondrial proteins are distributed differently between the two fractions.

Subcellular fractions were obtained from rat liver homogenates under conditions which prevented degradation of polysomes (pH 8.5 and high ionic strength). Rough endoplasmic reticulum (RER) was recovered in high yields from a low-speed nuclear pellet (rapidly sedimenting endoplasmic reticulum, RSER) and from a postmitochondrial supernate (rough microsomes). The polysomal RNA content of these two fractions was very similar. When polyA+-RNA's were translated inthe mRNA-dependent wheat embryo cell-free system, both fractions yielded polypeptide products which had similar electrophoretic patterns on sodium dodecyl sulfate (SDS)-polyacrylamide gels. Activities of messenger RNA's which code for albumin and for polypeptides destined for transport to the inner membrane and matrix of mitochondria (i.e. 'mitoplasts') were assayed by translating in the more active rabbit reticulocyte cell-free system followed by immunoprecipitation of radioactive products and coelectrophoresis with immunoprecipitated marker proteins on SDS-polyacrylamide gels. These tests indicated that albumin mRNA is about equally distributed between the two fractions of RER, or slightly enriched in the RSER fraction when activity is expressed as a percentage of total polypeptide synthesis. Activities of cytoplasmic mRNA's which code for at least some mitoplast proteins could be detected in both fractions, but all were enriched in the rough microsome fraction, not the RSER (two- to threefold when corrected for differences in total polypeptide synthesis in the lysate). Comparisons of mRNA's from free vs. membrane-bound polysomes indicated that most of the albumin mRNA activity (86-91%) and mitoplast protein mRNA activities (75%) were present in the bound fraction. Assuming that RSER and rough microsomes do not derive exclusively from different cells types, the evidence suggests that, compared to albumin and most other membrane-bound mRNA's, cytoplasmic mRNA's coding for mitoplast proteins may be preferentially segregated or compartmentalized within the cell on the microsomal class of RER.

Albumins

Subcellular fractionation of the porcine corpus luteum: sequestration of progesterone in a unique particulate fraction.

Homogenates of porcine corpus luteum were subjected to fractionation by differential-rate centrifugation or sucrose density gradient fractionation, with or without pretreatment with digitonin. Fractions of each gradient were assayed for a number of markers characteristic of the major intracellular organelles and cell-surface membranes, and for progesterone content. The majority of the progesterone content of homogenates of porcine corpus luteum was associated with a low-density particulate fraction which equilibrated at a buoyant density of 1.07-1.09 g/cm3. Pretreatment with digitonin increased the buoyant density of the progesterone-enriched fraction markedly (to 1.13-1.15 g/cm3) without causing release of steroid. The density distributions of progesterone content in control and digitonin-treated luteal gradient fractions were quite distinct from those of the major intracellular organelles and luteal cell-surface membranes. However, NADH-cytochrome C reductase activity (but not other endoplasmic reticulum markers) was also enriched in this fraction. The results suggest that most of the progesterone of the porcine corpus luteum is associated with a unique particulate fraction which is enriched in digitonin-reactive lipids and NADH-cytochrome C reductase activity.

Animals

Variations of the hypoxic fraction in the SCC VII tumors after single dose and during fractionated radiation therapy: assessment without anesthesia or physical restraint of mice.

Variations of the hypoxic fraction (HF) after single dose (13 Gy or 4 Gy) and during fractionated (5 fractions of 4 Gy, 1 or 2 fractions per day) radiation therapy were studied in SCC VII tumors implanted subcutaneously in the hind legs of C3H/He/Jms mice using the paired survival curve method. Whole-body irradiation was delivered to tumor-bearing mice without anesthesia or physical restraint, because both are known to increase the HF artificially. The HF decreased after a single 13 Gy dose in a biphasic fashion: extremely rapidly within 1 hr and comparatively slowly during the following 12-72 hr. On the other hand, nearly no fall of HF was observed in 24 hr following a single 4 Gy dose. Also, reoxygenation was found to occur more rapidly in the interfraction period as the number of fractions of 4 Gy increased irrespective of differences of interfraction time. However, the HF just before each radiation fraction was significantly higher than the pretreatment level for both fractionated regimens. Thus, the reoxygenation patterns observed after single low and high doses of irradiation were different from each other, and reoxygenation in each interfraction period did not always proceed in a similar manner to that after single low dose irradiation. Reoxygenation was facilitated as fractionated radiation therapy proceeded, but it was not sufficient for the HF to remain at a level comparable to that before irradiation.

Anesthesia

Elective lower neck irradiation: 5000 cGy/25 fractions versus 4050 cGy/15 fractions.

This is an analysis of 171 patients treated with irradiation alone for squamous cell carcinoma of the head and neck who received elective radiation therapy to the clinically negative low neck. All patients have a minimum follow-up of 2 years and 63% have had at least 5 years of follow-up. Patients were excluded from this analysis if they developed recurrent disease at the primary site or in the upper neck. The majority of patients, 162/171, were treated with one of 2 dose-fractionation protocols: 5000 cGy/25 fractions (5 fractions per week for 5 weeks) and 4050 cGy/15 fractions (5 fractions per week for 3 weeks). Patients were subdivided into four risk groups based on the estimated likelihood of subclinical disease in the neck. There was no difference in the rate of the neck disease control between these 2 dose-fractionation protocols. We conclude that 5000 cGy/25 fractions and 4050 cGy/15 fractions are equally effective in sterilizing occult neck disease.

Carcinoma, Squamous Cell