A genome-wide screen in Saccharomyces cerevisiae for genes affecting UV radiation sensitivity.

The recent completion of the deletion of essentially all of the ORFs in yeast is an important new resource for identifying the phenotypes of unknown genes. Each ORF is replaced with a cassette containing unique tag sequences that allow rapid parallel analysis of strains in a pool by using hybridization to a high-density oligonucleotide array. We examined the utility of this system to identify genes conferring resistance to UV irradiation by using a pool of 4,627 individual homozygous deletion strains (representing deletions of all nonessential genes). We identified most of the nonessential genes previously shown to be involved in nucleotide excision repair, in cell cycle checkpoints, in homologous recombination, and in postreplication repair after UV damage. We also identified and individually confirmed, by replacing the genes, three new genes, to our knowledge not previously reported to confer UV sensitivity when deleted. Two of these newly identified genes have human orthologs associated with cancer, demonstrating the potential of this system to uncover human genes affecting sensitivity to DNA-damaging agents and genes potentially involved in cancer formation.

Functional characterization of the S. cerevisiae genome by gene deletion and parallel analysis.

The functions of many open reading frames (ORFs) identified in genome-sequencing projects are unknown. New, whole-genome approaches are required to systematically determine their function. A total of 6925 Saccharomyces cerevisiae strains were constructed, by a high-throughput strategy, each with a precise deletion of one of 2026 ORFs (more than one-third of the ORFs in the genome). Of the deleted ORFs, 17 percent were essential for viability in rich medium. The phenotypes of more than 500 deletion strains were assayed in parallel. Of the deletion strains, 40 percent showed quantitative growth defects in either rich or minimal medium.

COP1, an Arabidopsis regulatory gene, encodes a protein with both a zinc-binding motif and a G beta homologous domain.

Plant seedling development is capable of following 1 of 2 distinct morphogenic pathways: skotomorphogenesis in darkness and photomorphogenesis in light. Dark-grown Arabidopsis seedlings with recessive mutations at the constitutively photomorphogenic (COP1) locus indicate that the wild-type COP1 protein represses photomorphogenesis in darkness and that light reverses this repressive activity. Using a T-DNA-tagged mutant, we have cloned the COP1 locus. The amino-terminal half of the encoded protein contains a conserved zinc-binding motif, whereas the carboxyl-terminal half contains a domain homologous to the WD-40 repeat motif of G beta proteins. The presence of both a putative DNA-binding motif and a G protein-related domain in a single polypeptide suggests that COP1 may be the first of a new class of regulatory molecules. This novel structure could endow COP1 with the capacity to function as a negative transcriptional regulator capable of direct interaction with components of the G protein signaling pathway.

Radiation therapy of seminoma of the testis.

This study is a review of 66 patients with a diagnosis of testicular seminoma, treated at the Department of Radiation Oncology, University of Louisville, from 1959 to 1978. Sixty-four of the patients were diagnosed as pure seminomas and two as spermatocytic type. The age of the patients ranged from 16 to 75 years with a median of 39. Fifty-four patients (82%) presented with swelling or mass in the testis, 7 patients with pain, 1 patient with hydrocele, and 4 patients had seminoma diagnosed incidentally. The distribution was equal for both right and left sides. There were 39 (59%) and 27 (41%) patients in stages I and II, respectively. All stage I patients received only infradiaphragmatic irradiation by a "hockey stick" field, and the majority of them received a dose of 3,200-3,600 rads in 3-4 weeks time. Stage II patients received elective irradiation to the mediastinum and neck region, in addition to the infradiaphragmatic irradiation. The 5-year actuarial survival rates for stage I and II seminomas are 96% and 92%, respectively. There were no serious complications during follow-up. The role of elective irradiation in stage I disease is discussed with a brief review of the literature.

Non-metastatic locally advanced cancer of the breast treated with radiation.

One hundred forty-seven female patients with retrospectively classified (AJCCS 1977) Stage III and nonmetastatic noninflammatory Stage IV breast carcinoma from June 1958 to December 1978 were studied to determine the effects of primary radiation on local recurrence and survival. Fifty-one patients recurred in the breast; 24 patients recurred in the local-regional area of breast and draining lymphatics and 15 patients recurred in the axilla only. Distant metastases developed in 97 patients. The 5 year actuarial survival was 24%. Decreased recurrence rates were associated with megavoltage and higher doses of radiation (34% vs. 65%); with smaller tumors (27% vs. 57%); with total extirpation of gross tumor (33% vs. 53%); and with those tumors that were given a "boost dose" (26% vs. 58%). The addition of systemic treatment did not appreciably alter either local-regional recurrence or survival. However, most of the patients received endocrine ablation or single-agent chemotherapy. Polyagent chemotherapy was used late in the series, making its true impact on survival difficult to evaluate.

Irradiation of nasopharyngeal carcinoma: correlations with treatment factors and stage.

Eighty patients with nasopharyngeal carcinoma were treated with radiotherapy in the Radiation Center at the University of Louisville from January 1955 to December 1980. Among the patients were 70 whites, nine blacks and one Chinese; their ages ranged from eight to 82 years. There was a 40% recurrence rate within the nasopharynx, and a 29% recurrence rate within neck nodes. The five year survival and relapse-free survival rates of the entire group were 36 and 33%, respectively. Forty-nine patients died of cancer, four patients died of intercurrent disease and eight patients were lost to follow-up. Nineteen patients are alive and free of disease. Factors considered in this study included tumor and nodal status, the presence of cranial neuropathy, the size and area irradiated, and dose delivered. Primary site relapse was not demonstrated to be dependent on T group or nodal status, but was likely to be related to inadequacy of original treatment volume and dose. A higher survival was noted with our lymphoepithelioma category (p = .056).

Survival and recurrence patterns in the radiation treatment of carcinoma of the vagina.

Fifty-three patients with invasive cancer of the vagina were treated with a curative course of radiation at Tufts New England Medical Center between September 1958 and December 1979. There were 37 patients with "primary" carcinoma of the vagina and 16 with metastatic disease to this site. The treatment comprised an integration of teletherapy and intracavitary/interstitial therapy. The 5-year relapse-free survival for primary vaginal carcinoma was 88% for Stage I, 44% for Stage II, 35% for Stage III, and 0% for Stage IV. Patients with metastatic disease had an overall 5-year survival of 63%. There was local recurrence in 35% of the patients with primary and 19% with metastatic vaginal carcinomas. The incidence of local recurrence appeared to be increased with higher grade pathology, lesions other than vault carcinomas, and lesions involving the entire length of the vagina. Due to immediacy of adjacent structures in the high dose volume, complications excluding those patients with persistent recurrent disease were somewhat high (11%).

Dysgerminoma of the ovary.

Dysgerminomas of the ovary are rare germ cell tumors. They are more common at puberty. Surgery followed by irradiation has resulted in excellent cure rates in early stages of this disease. This paper is a review of six patients with a diagnosis of dysgerminoma who were treated at the Department of Radiation Oncology, University of Louisville. The controversy on the use of irradiation in Stage Ia tumor is discussed in detail. A review of the literature is also done in this paper.

Patterns of local-regional recurrence and results in Stages I and II breast cancer treated by irradiation following limited surgery. An update.

One hundred forty-six women with Stage I and Stage II breast cancer received radical radiotherapy after having excisional biopsy ( lumpectomy ) at Massachusetts General Hospital between 1956-1978. They were grouped according to age: those younger than 49 years and those older than 50 years. The 5-year survival rates were 93 and 73% for patients with Stage I and Stage II cancer, respectively; the corresponding 5-year relapse survival rates were 75 and 56%. The local recurrence rate was 8% in patients with Stage I disease and 17% in those with Stage II disease. Survival was not significantly affected by patients' age, by the presence or absence of blood vessel or lymphatic involvement, or by the addition of adjuvant chemotherapy. No major complications occurred. Modification in radiation dose and technique resulted in improved overall survival and local control. Limited surgery followed by radical radiation therapy offers a therapeutically effective, cosmetically acceptable alternative to radical surgery for early stage breast cancer.

Is mediastinal irradiation necessary for stage I testicular seminoma?

This study is a review of 39 patients with testicular seminoma, Stage I, treated at the Department of Radiation Oncology, James Graham Brown Cancer Center from 1959 to 1978. The age of the patients ranged from 16 to 70 years with a median of 37. Thirty-two (82%) patients presented with swelling or mass in the testis, four patients with pain, and three patients had seminoma diagnosed incidentally. Twenty (51%) patients had the tumor on the right side and 19 (49%) patients had the tumor on the left side. All patients received irradiation to the ipsilateral inguinal, iliac, and bilateral para-aortic nodes with "hockey stick" type fields. The majority of the patients received a midplane dose of 3,200 to 3,600 rad in 3-4 weeks time. None of the patients received prophylactic irradiation to the mediastinum and supraclavicular region. The 5-year actuarial survival rate is 96%. There is no additional benefit in giving prophylactic irradiation to the mediastinum and supraclavicular regions in Stage I testicular seminoma. A brief review of the literature regarding the role of prophylactic irradiation in this group of patients is done.

Posthysterectomy megavoltage irradiation in the treatment of cervical carcinoma.

Current literature has demonstrated excellent results using postoperative megavoltage irradiation in patients who had simple hysterectomy in the presence of invasive cervical carcinoma. It has recently been suggested that total abdominal hysterectomy followed by postoperative irradiation should be used as a planned treatment for early-stage cervical carcinoma. Thirty-six cases of patients with cervical carcinoma treated with whole-pelvis irradiation following simple hysterectomy at the University of Louisville are reviewed.

Solitary plasmacytoma.

A retrospective review is presented of nine patients presenting with solitary plasmacytoma. Between 1963 and 1980, these patients were treated with radiation at the Department of Radiation Oncology, University of Louisville. Criteria for admission to this particular study include (1) a solitary focus of plasmacytoma proven by biopsy, (2) normal bone marrow findings (less than 10% plasma cells), and (3) no evidence of disseminated disease. In six patients the primary site was osseous, and in three extramedullary, two of which were located in the nasopharynx and nasal cavity and the third in the stomach. All of the extramedullary plasmacytomas are disease free for periods ranging from 4 to 10 years. Of the six patients with osseous lesions, two developed multiple myeloma in 2 and 3 years, two are NED (No Evidence of Disorder) after 9 years, one died of intercurrent disease, and the remaining patient was NED for 2 years, after which he was lost to follow-up. These results suggest the more favorable prognosis of extramedullary plasmacytoma and support the theory that the solitary plasmacytoma of bone and extramedullary plasmacytoma are distinct disease entities.

Computed tomography and radiotherapy in the treatment of cancer.

Computed tomography (CT) has shown to be of great value in the treatment of cancer with radiation therapy. It is used more and more in the estimation of tumor volume and for treatment planning, with the aid of the computerized treatment unit. At the James Graham Brown Cancer Center, Department of Radiation Oncology, CT has been used routinely for the treatment planning. From October 1, 1981 to June 30, 1982, we performed 180 CT scans for the treatment planning, 380 simple dose calculations, 237 complex treatment plans, and 42 intracavitary dosimetry using the treatment planning unit. This is a review of our experience with some illustrations. Accurate tumor dose can be delivered with reducing the complications with the use of CT and the computerized treatment unit.

Effect of whole-body hyperthermia on cell survival, metastasis frequency, and host immunity in moderately and weakly immunogenic murine tumors.

The effects of whole-body hyperthermia (WBH) on animal tumors and on metastasis frequency were studied. The tumors were a chemically-induced fibrosarcoma, FSa-I, which is moderately immunogenic and a spontaneously arisen fibrosarcoma, FSa-II, which is very weakly immunogenic. The WBH was given at 42.5 degrees in an incubator which had an auxiliary heater for accurate temperature control. Animal core temperature reached 41.5 degrees in 30 min. The lung colony assay revealed that the WBH for 60 min given at 24 hr after i.v. injection of single cells gave no lethal damage to either FSa-I or -II tumor cells. A significant inhibition of tumor growth was found when large tumors were given three daily WBH treatments. The frequency of lung metastasis was enhanced when large weakly immunogenic FSa-II tumors were treated by WBH, although no increase in the frequency was observed for FSa-I tumors of any size. Local hyperthermia did not significantly increase the metastasis rate of both tumors. These results suggest that the WBH might be useful for a treatment of large immunogenic tumors. However, the WBH is not a choice of treatment for possible micro-metastases.

Comparison of adjuvant postoperative radiotherapy and multiple-drug chemotherapy (CMF-VP) in operable breast cancer patients with more than four positive axillary lymph nodes.

With the advent of the adjuvant chemotherapy in 1974 for early operable breast cancer at Massachusetts General Hospital, 41 patients with greater than or equal to 4 lymph nodes positive in their axillary dissection received a two-year course of the modified Cooper regimen (S + CH) between March 1974 and June 1976. The actuarial disease-free survival was found to be comparable to CMF as reported by Bonnadonna. No consistent randomization with a control group was possible. Since postoperative radiotherapy has been proven to decrease the incidence of local recurrence but does not affect survival; it was believed that this group should serve as an adequate control. Sixty patients with greater than or equal to 4 positive axillary lymph nodes in their axillary dissection who were given postoperative radiotherapy to the chest wall and draining lymphatics (S + RT) in 1973 and 1974 at the same institution were retrospectively analyzed and compared to the above group. Relatively little significant difference in survival and disease-free survival curves was found. At 48 months, there was 72% and 58% survival for the S + RT and S + CH groups, respectively. When subgrouped according to age, those patients less than 49-years-old had a 48-month disease-free survival of 65% and 49% for S + RT and S + CH groups, respectively (P = 0.25). No significant difference was noted in the post menopausal group. In both groups, patients with greater than 50% positive axillary lymph nodes exhibited a poor prognosis.

Effect of X-rays and cyclophosphamide on solid tumors and naturally occurring metastases in mice.

Combinations of single doses of X-rays given locally to a transplanted tumor and single i.p. doses of cyclophosphamide (CY) were tested in mice bearing solid transplanted fibrosarcomas. The end points examined were growth delay and local control of the implanted tumor, the incidence of distant metastases, and survival times free of either local recurrence or metastases. Growth delay and local control increased more steeply with X-ray dose than with dose of CY. From 8 to 10 days of extra regrowth delay required only 10 to 20% extra X-ray dose, but it required a doubling of CY dose from three-eighths to three-fourths of the maximum tolerated dose. The incidence of metastases and the survival time of the mice also depended more upon the local dose of X-rays than that of CY. This result was not expected and suggests that metastases are eliminated more certainly if the primary (implanted) tumor is locally controlled on a long-term basis. A significant proportion of mice, over 50%, was cured of both the transplanted tumor and distant metastases only when the highest doses of both X-rays and CY were given simultaneously. Extended intervals between the two agents gave inferior results, intervals of 8 days giving significantly worse results, but 4 days giving not significantly worse results than simultaneous administration, especially when the X-ray treatment was given first. The interval of 4 days would, however, be sufficient to avoid the enhancement of radiation injury in normal tissues in mice.

Effect of cyclophosphamide or x-rays on spontaneously occurring metastases from tumors transplanted into the tails of mice.

A spontaneous metastases model in mice is being used to test the efficiency of various treatments in eliminating metastases. Solid tumors were transplanted into the tails of mice and removed by tail transection when they had grown to a 4- to 5- or 6- to 7-mm mean diameter. Subsequently, 70 to 95% of mice not given other treatment developed metastases in the lungs or in regional lymph nodes (lumbar sacral region), or in both sites. The present paper reports the effects of whole-body or partial-body treatment on these metastases. The treatments, which started at the time of surgical transection of the tail, included a range of single or fractionated doses of cyclophosphamide (CTX) or X-rays given either to the whole body or locally to the lungs only. CTX reduced the incidence of metastases in both sites although the incidence of lung metastases was reduced by smaller doses of CTX than that of the lumbar sacral metastases. Whole-body irradiation of 6 grays (600 rads) had no effect on the incidence of metastases, whereas local irradiation of the lungs with single doses of 14.5 or 20 grays reduced the number substantially, as did 95 mg or more of CTX per kg. Thus, CTX or radiation reduced the incidence of lung metastases in a system where metastases developed from cells seeded from a primary tumor rather than from a cell suspension injected into the tail vein.

Inflammatory breast carcinoma treated by radical radiotherapy.

Sixty-two patients with localized clinical inflammatory breast carcinoma were treated with curative, radical radiotherapy doses to the breast and draining lymphatics at Massachusetts General Hospital from January 1960 to December 1977. Fifty patients have died from disease, 7 are alive with disease, and 5 are free of disease at time of reporting, thus indicating the fulminant nature of this rare form of breast cancer. Median survival is approximately 18 months (mean 24 months). A five-year actuarial survival relapse-free survival of 14% and 6%, respectively, are obtained. Local and regional recurrence was noted in 43 of 62 patients (69%). Twelve patients (19%) failed in the locally irradiated area only, 31 patients (50%) failed in both local, regional, and distant sites, and 14 patients (23%) failed with distant metastases only. Due to the complex dosimetry required in the treatment of breast cancer, retrospective analysis was made of actual tumor doses delivered before 1972. The breast was oftentimes calculated to receive 20 to 35% less than the stated dose. This gave the unique opportunity of observing a dose response relationship in local control within the same institution. Doses in excess of 6000 rads tumor dose seem necessary since 14 of the 15 patients with persistent disease had received less than this dose. However, once tumor exceeded 10 cm, increasing dose within clinical therapeutic ranges failed to control disease, although the recurrence-free interval was somewhat prolonged. Since 1976, twice-a-day fractionation has been used in larger tumors, and this appears to have decreased the local recurrence rate to 33% (2/6) patients. Preliminary results of adjuvant multiple drug therapy, e.g., modified Cooper regimen (CMFVP) or the CMF regime appear encouraging.