A retrospective study of follicular lymphomas.

Sixty-four cases of follicular lymphoma followed-up for 10 years have been studied. The influence of age, sex, histological appearance, immunophenotype and T-cell content on prognosis has been examined. Initial evaluation indicated that increasing age, relatively low numbers of intrafollicular T-cells and absence of mantle zones around the neoplastic follicles were associated significantly with mortality. Multiple regression analysis with adjustment for age and sex was performed and the only significant variable was then found to be the histological grade.

Charged particle irradiation of chordoma and chondrosarcoma of the base of skull and cervical spine: the Lawrence Berkeley Laboratory experience.

Forty-five consecutive patients with chordoma or chondrosarcoma at the base of skull or cervical spine were treated at the University of California Lawrence Berkeley Laboratory (UCLBL) and University of California School of Medicine, San Francisco (UCSF) between November 1977 and October 1986. All patients had undergone a subtotal surgical resection. Twenty-three patients were treated definitively with charged particles, 13 patients were treated with photons and particles, and 9 patients were treated for recurrent disease. Total doses ranged from 36 to 80 Gray equivalent (GyE). Thirty-three patients are alive with a minimum followup of 1 year. The actuarial survival and local control for all patients at 5 years is 62% and 59%, respectively. Patients treated for primary disease had a 78% actuarial local control rate at 2 years, whereas the rate for patients with recurrent disease was 33%. Patients with smaller visible tumor volumes (less than 20 cc) had a significantly better local control rate than patients with larger tumor volumes (80% vs 33% actuarial rate at 5 years). Patients with chondrosarcoma had the highest local control rate, as did patients treated with particles alone. Complications included 3 patients with unilateral visual loss, two patients who became blind, and 4 patients with radiation injury to the brainstem.

Southern blot analysis of DNA extracted from formal-saline fixed and paraffin wax embedded tissue.

Model experiments were designed to assess whether DNA could be recovered from formol-saline fixed peripheral blood lymphocytes and tonsil tissue for use in Southern blot gene analysis. Lymphocytes were fixed for 30 min and tonsil for 6 and 24 h, then paraffin embedded. High molecular weight DNA was extracted by prolonged digestion (2-7 days) with proteinase K or protease XXIV in the presence of 1 per cent sodium dodecyl sulphate. Restriction, transfer and hydridization were possible without modification of standard procedures. Multiple copy sequences were demonstrated using Mspl and Bst Nl restriction and hybridization for the Y chromosome (pHY 2.1 probe), single copy genes using EcoRI and BamHl restriction for the T-cell receptor beta chain (T beta probe), and Bgl II and Hind III for the immunoglobulin heavy chain (JH probe). Identical banding to unfixed tissue was achieved except when 24 h fixed extracts were used. With these, demonstration of the 24 KB Bam Hl/T beta and 9.2 KB Hind III/JH bands was not obtained. These findings suggest that as the fixation time is extended, alterations to DNA will limit the available range of restriction enzyme/probe combinations. However, with careful choice of these the extraction of DNA from formalin fixed and paraffin embedded pathological tissue for Southern blotting should be profitable.

Dosimetry considerations for a Lipowitz metal tissue compensator system.

The present report describes the fabrication technique and the dosimetry aspects of a new compensator system that uses the low melting point allow Lipowitz metal. Compensating ratios (CR, mm of tissue compensated per mm of cerrobend) were determined for various field sizes and depths for 60Co, and X ray energies of 4, 6, 18, and 25 MV. Typical CR for 10 cm X 10 cm field and 10 cm depth were: 60Co, 4 MV, and 6 MV, 1:15; 18 and 25 MV, 1:20. Verification of these CR were performed at 6 MV using polystyrene phantoms. Ionization measurements were made for various field sizes and depths and normalized to central axis full-phantom readings for both compensated and non-compensated fields. Without compensation, percent differences ranged as high as 40% for a tissue deficit of 10 cm. With the compensating filters (CF) in place, this difference was reduced to 2-4%. Overall, the CF system was capable of producing dose uniformity to within +/- 10% for a wide range of depths and field sizes.

Comparison of cancer chemotherapeutic agents in asynchronous and synchronous 9L cells.

The cytotoxic activity of various chemotherapy agents was investigated in asynchronous populations of cultured 9L rat brain tumor cells, and as a function of their position in the cell cycle. Representative drugs from the classes of DNA-active agents, alkylating agents, spindle poisons, and antimetabolites were tested. The ability to induce cell lethality in asynchronous populations as a function of drug concentration varied for 1 hr pulse exposures. In order of decreasing cytotoxic activity, DHAQ was the most effective, followed by VCR, VDS, VBL, ADR, BCNU, cis-DDP, BLM, DBD, RZ, and HU. The effect of chemotherapy agents on synchronous 9L cells obtained by mitotic selection also varied with respect to the individual agent and was cell cycle-dependent. Survival age-responses ranged from being minimal to demonstrating significant fluctuations as a function of cell cycle position. For all agents except ADR and HU, the sensitivity of G1 phase was greater than S phase. RZ exhibited essentially a flat age-response. Comparison of the cell cycle age-responses of chemotherapeutic agents to those exhibited by the cytotoxic modalities of radiation and hyperthermia demonstrate several unique differences.

Dosimetry of CT-guided volumetric IR-192 brain implant.

Over the past 2 years, an afterloading technique has been developed and refined to implant radioactive Ir-192 sources into brain tumors. The implantation procedure integrates a stereotaxic system with computerized tomography (CT), which provides tumor position, volume, and guides the placement of catheters. A radiolucent ring-frame immobilizes the head as holes are made at 1 cm intervals with the aid of a template. Catheters containing dummy sources 1 cm apart are then inserted to the desired depth, and their position verified in three dimensions to insure complete coverage of visible tumor volume as defined by contrast enhancement. Once catheters are secured, the anesthetized patient is moved to the intensive care unit where the dummy sources are replaced by ribbons of Ir-192 seeds (specific activity 0.6-1.0 mg Ra eq). CT scans with the dummy sources in place are used to designate spatial coordinates of the active sources. A computer program converts position data and source strength into isodose contours in any plane. The implant duration (70-100 hours) for the desired dose to the tumor periphery (60-120 Gy) is then calculated. Dose rate contours are superimposed on preimplant CT scans. Maximum and minimum doses are determined in each of the various planes. Verification dosimetry has been carried out with thermoluminescent dosimeters placed in a catheter located in a plane along the tumor periphery. In vivo isodose values compared to idealized plans agree within +/-5%-10%.

ARH-77, an established human IgG-producing myeloma cell line. II. Growth kinetics, clonogenic capacity, chalone production, xenogeneic transplantations, and response to melphalan.

The growth curve of monolayer cultures of ARH-77 cells, a human myeloma cell line propagated in vitro, is represented by an everbending curve on a semilogarithmic plot; however, the curve can be fitted by a straight line on a linear-linear plot. This unusual growth pattern suggests that, instead of a fixed proportion of the population, a fixed number of ARH-77 cells divide per unit time. The following are cell cycle transit time parameters calculated from percent labeled mitosis experiments: TG1, 10.0 +/- 3.5 hours; Ts, 14.3 +/- 2.3 hours; TG2, 4.3 hours; TM, 1.4 +/- 1.3 hours; and Tc, 30.0 +/- 6.1 hours. For cells exposed continuously to 3H-thymidine the values are: growth fraction, 67%; TG1, 6.5 hours; Ts, 13.0 hours; and TG2 + M, 3.0 hours. The average doubling time is 4.6 days (range, 3.8-4.7 days); after about 10 to 15 days in culture, the growth rate of freshly passaged cells declines markedly, as reflected by a growth curve with a much shallower slope. The changes are accompanied by a marked decline in the labeling index from 41.3% (range, 28.9%-53.7%) during the first 3 days of culture to less than 5% measured on day 21. Flow cytometry for DNA content-dependent cell cycle compartment distribution demonstrates an obvious decline in the proportion of S-phase cells and a marked accumulation of G2 phase cells as the cultures age. When the supernatant medium of ARH-77 cells grown for 10 days is replaced by fresh medium, a new burst of vigorous cellular growth is observed with a curve slope similar to that observed during the first 5 days of culture. If the 10-day-old supernatant medium is used to set up cultures with freshly harvested ARH-77 cells, their growth curve resembles that of 10-day-old cultures. However, this supernatant medium induces no decrease in the growth rate of other human tumor cells, suggesting that inhibition of cellular growth does not result from exhaustion of nutrients, but that ARH-77 cells produce a molecular mediator that specifically inhibits the growth of these cells. ARH-77 cells could be synchronized with a single treatment of 3 or 5 mM thymidine; (dThd) and cloning efficiency was 2% to 4% in a double-layer soft agar assay. Treatment for 1 hour with increasing concentrations of melphalan produced a threshold exponential survival curve (Dq = 0.45 microgram/ml and D0 = 0.35 microgram/ml, 1 hour).(ABSTRACT TRUNCATED AT 400 WORDS)

Development of normal tissue damage in the rat subsequent to thoracic irradiation and prior treatment with cancer chemotherapeutic agents.

The effect of combined modalities (radiotherapy-chemotherapy) on the development of long-term normal tissue damage was investigated in rats. Animals received single I.P. injections of Hank's balanced saline solution, adriamycin (ADR, 1.0 mg/kg), bleomycin (BLM, 10 units/kg), or dihydroxyanthraquinone (DHAQ, 3.0 mg/kg); and/or irradiation of the chest with 25 MV x-rays (12 Gy) at 0, 43, 93, or 199 days after drug treatment. Only animals treated with DHAQ displayed appreciable toxicity, with more animals dying at less than 200 days when radiation was added at 0 or 43 days. Although animals treated with BLM or radiation exhibited evidence of lung damage (histologically by 199 days and radiographically by 300 days), their survival was not compromised. The simultaneous administration of x-ray and BLM produced enhanced effects as compared to either agent alone. These results demonstrate an enhancement of normal tissue damage by combined treatment with radiation and chemotherapeutic agents, not only for acute toxicity but also for long-term effects. This damage was ultimately expressed as alteration of lung structure (histologically and radiographically) in the case of BLM, and as animal lethality in the case of DHAQ. In addition, there was a reduction in the degree of enhancement observed as a function of the separation in time between treatment with chemotherapeutic agents and subsequent irradiation. These factors should be considered when combined modality therapy is used for treatment of cancer in the thoracic region.

Angiotensin-I-converting enzyme in cancer patients.

Serum levels of angiotensin-I-converting enzyme (ACE) were assayed before, during, and after radiation therapy in 209 patients receiving treatment for neoplastic disease. Daily fluctuations in the measured ACE levels were minimized by comparing all patient values to that of a simultaneously run sample from a standard source of serum obtained by pooling sera from young, healthy volunteers. Most of the patients tested presented with a normal to low ACE level, with the mean value for all patients being 70% of the standard value. More patients with primary lung cancer displayed values that were low (107 of 120) than did patients with disease outside the lung (44 of 78), this difference being statistically significant at p less than 0.001. In addition, more patients with lung cancer had values less than 70% of the standard than did patients with disease outside the lungs. These initial results suggest that monitoring of serum ACE levels may be useful in the management of patients with malignant disease in the lung.

Effect of dihydroxyanthraquinone on the response of a rat solid tumor to radiation therapy.

Dihydroxyanthraquinone (DHAQ, NSC 279836, a new cancer chemotherapeutic agent presently in clinical trials) was tested, alone and in combination with radiation, for therapeutic effectiveness in a rat solid tumor model. The Walker 256 fibrosarcoma was implanted subcutaneously in the leg via trochar so as to yield palpable tumors within 7-11 days. When tumor diameters reached 1 cm, the animals were treated as follows: as controls; with DHAQ (3.0 mg/kg, i.p.); with 300 KVP x-rays (15 Gy to the tumor-burdened leg); or with a combination of DHAQ and radiation. Tumor diameters were measured three times a week to monitor the response to therapy. DHAQ alone had no effect on tumor growth rate. Radiation alone produced a delay in tumor growth. The combination of DHAQ and radiation resulted in a consistently better therapeutic response than did radiation only, but at only a marginal level of statistical significance (0.05 less than p less than 0.10). However, there were 3/10 long-term survivors (greater than 80 days post treatment) without evidence of tumor in the combination therapy group; whereas all animals in the radiation group died or were sacrificed because of progressive tumor growth by day 71. These results suggest that even though DHAQ is only minimally effective against this rat solid tumor when used alone, there may still be some added therapeutic benefit derived from a combination of DHAQ and radiation therapy.

Bleomycin and brain tumors. A review.

A logical inference from the recent reports indicating that malignant brain tumors are composed of a heterogeneous cell population is that combination chemotherapy will be required for effective brain tumor control. For several years we have been investigating the use of Bleomycin as an agent to be used in conjunction with radiation therapy and a nitrosourea compound. Since systemically administered Bleomycin does not cross the blood-brain-barrier and has significant toxicity when used parenterally in high doses, we have studied the use of smaller doses of Bleomycin injected directly into the brain tumor cavity. Such an intracerebral dose was more effective in prolonging survival of rats burdened with experimental 9L gliosarcomas than an intravenous dose that is 25 times as great. The combination of intracerebrally administered Bleomycin and radiation therapy was more effective than either modality alone. Furthermore, the combination of Bleomycin delivered intracerebrally and BCNU given systemically was more effective than either agent used alone. Finally, in a Phase I clinical of Bleomycin given via an Ommaya reservoir to eight patients with recurrent malignant brain tumors, we have demonstrated that individual doses of up to 7.5 units and cumulative doses of up to 255 units can be administered without significant toxicity.

Growth characteristics of human colonic adenocarcinomas propagated in the Rowett athymic rat.

Two human colonic adenocarcinoma cell lines were propagated in the Rowett athymic rat. Line LoVo displayed a well-differentiated morphology and exhibited progressive growth over a 70-day observation period with a doubling time of 8.5 days throughout. No metastatic involvement was noted. Xenografts of SW 620 cells were undifferentiated and highly necrotic. These tumors grew progressively for approximately 30 days with a doubling time of 5.5 days, but over 90% of the animals exhibited a spontaneous regression with a mean time to total regression of 51 +/- 3 (S.E.) days. Animals which had rejected the SW 620 xenografts would not support the growth of either the SW 620 or LoVo xenografts when challenged a second time with inocula producing a 100% tumor incidence in control rats. No metastatic involvement was noted for the SW 620 xenografts, but the tumors frequently invaded the underlying musculature and protruded into the peritoneal cavity without producing ascites or tumor nodules at sites distant from the primary.

Effect of dihydroxyanthraquinone (NSC 279836) and thoracic irradiation on long-term survival of rats.

Dihydroxyanthraquinone (DHAQ; NSC 279836) is a recently synthesized compound that is structurally similar to Adriamycin and produces greater antitumor effects in murine model systems. We compared DHAQ to Adriamycin in rats, with and without irradiation of the chest at various intervals after drug treatment. A single injection of Adriamycin (1 mg/kg i.p.) had little effect on animal survival, even if combined with radiation (12 Gy 25 MV X-rays), greater than 90% being alive at 1 year. A single injection of DHAQ (3 mg/kg i.p.) was equally uneffective up to 200 days after treatment (survival, greater than 90%). However, between 200 and 370 days after treatment, all animals died, producing a median survival time of 280 days. Further, when DHAQ was combined with radiation, there was an increase in animal deaths between Days 300 and 200. For animals irradiated on Days 0, 43, and 93 after DHAQ treatment, only 50, 75, and 80%, respectively, survived to Day 200. All animals that survived past Day 200 subsequently died by 1 year, displaying the same kinetics of lethality as those animals that had received DHAQ only. A repeat experiment using DHAQ at 1 mg/kg produced similar results. Based on these findings, we conclude that DHAQ produces a long-term (greater than 200 days) toxicity in rats that is not detectable by short-duration toxicity screening. In addition, radiation enhances short-term (less than 200 days) lethality, with the degree of enhancement decreasing as the interval between drug and radiation is increased.