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Biomedical subjects

W A Brock

Publications and source records attributed to W A Brock.

At least 19 recordsLinked to original sources
PubMed1996

Cell density dependent plating efficiency affects outcome and interpretation of colony forming assays.

BACKGROUND AND PURPOSE: The usefulness of colony forming assays (CFA) has been established for almost 40 years (Puck and Marcus, J.Exp.Med. 103: 653-666, 1956). Although time-consuming and not successful for all cell lines, it is generally considered to be the gold standard of assays for testing the sensitivity of cell lines to ionizing radiation or other cytotoxic agents in vitro. We recently found for several cell lines that the plating efficiencies of both control and irradiated cells is dependent upon the density of cells seeded for colony formation; that is, increasing cell inoculum levels resulted in a non-linear relationship with colony formation, even at relatively low colony numbers. MATERIAL AND METHODS: All data from a human melanoma cell line, transfected with c-myc or N-ras, as well as from normal human diploid fibroblasts, were taken to see how this phenomenon influenced outcome and interpretation of clonogenic assays. Survival was recalculated using all data, or only data with a linear relationship between inoculum level and colony formation. RESULTS: It is found that when data with a non-linear relationship between inoculum level and colony formation are included, survival can be underestimated due to inhibition of colony formation in treated cultures. CONCLUSION: For validity, colony forming assays must be standardized to assure a constant relationship between the cell density and colony forming efficiency. This usually requires a much lower density of colonies than has been typically published for many cell survival studies.

Animals↗
PubMed1996

Repair of chromosome and DNA breaks versus cell survival in Chinese hamster cells.

Clonogenic and non-clonogenic parameters of cell survival were compared in irradiated Chinese hamster cells. Clonogenic survival, chromatid break and repair kinetics, as well as DNA damage and repair, were assessed in synchronized cells in different parts of the cell cycle. C2 chromatid damage and repair was examined in metaphase chromosomes of cells irradiated during S and G2 phase, treated with or without inhibitors of DNA repair. Bromodeoxyuridine labelling of S phase cells starting at the time of irradiation made it possible to determine precisely, while scoring metaphase chromosomes, whether cells were irradiated in mid S, late S, or G2 phases of the cycle. The results showed that chromatid breaks induced in S phase are efficiently repaired until the moment cells progress into G2, when repair stops abruptly. Chromatid damage in G2 phase is not repaired. On the other hand, DNA double-strand breaks are repaired in all phases of the cycle, even during G2 phase which has no concurrent chromatid break repair. Finally, there is no consistent correlation between chromatid damage and repair, DNA damage and repair, and cell survival, thus indicating that the interaction of different parameters of radiosensitivity must be better understood for them to be useful predictors of cell survival.

Animals↗
PubMed1995

Fibroblast radiosensitivity versus acute and late normal skin responses in patients treated for breast cancer.

PURPOSE/OBJECTIVE: To determine if the radiosensitivity of normal human skin fibroblasts, measured in early passage cultures, is significantly correlated with the degree of acute or late normal skin damage in patients treated for breast cancer with radiotherapy. METHODS AND MATERIALS: In the 1970s, a series of breast cancer patients was treated at the Department of Oncology in Gothenburg, Sweden with postoperative irradiation to the parasternal region. Patients were treated bilaterally using different fractionation schedules and doses to the right and left fields. Peak acute reactions were scored on a six-point scale, and skin erythema was measured by reflectance spectrophotometry. Telangiectasia was graded over time on a six-point scale. In April 1992, two small skin biopsies were obtained from 22 patients in two treatment groups (i.e., four dose-fractionation schedules) and, using either delayed or immediate plating, fibroblast radiosensitivity was measured in early passage cultures by clonogenic survival, after high and low dose-rate irradiations. Survival at 2.0 Gy (SF2) was calculated from complete survival curves. RESULTS: To test assay reproducibility, SF2 values derived from paired biopsies of the same patient (12 cases) were compared. A reasonably good correlation (p = 0.075) was obtained for SF2s determined by high dose-rate irradiations with immediate plating, but not for delayed plating or low dose-rate treatments. The median coefficient of variation in the replicate SF2s after high dose-rate treatment and immediate plating was 13%, suggesting that the poor correlation in paired SF2 values is due to the magnitude of the uncertainty in SF2 relative to the overall spread in SF2 values between patients (CV = 28%). Individual SF2 values and averaged values from patients with data from two biopsies were compared with the acute and late clinical reactions. A significant negative correlation was found between SF2 and relative clinical response, but only when averaged high dose-rate SF2 values and telangiectasia scores were compared. There was no significant correlation between average SF2 values and acute responses or between individual SF2 measurements and either the acute or late clinical response. CONCLUSION: The results of this study suggest that the degree of late telangiectasia is at least partially dependent upon the intrinsic cellular radiosensitivity of normal fibroblasts, but the relationship is not clear cut. Multiple replicate assays are necessary to obtain reliable estimates of fibroblast SF2 values using current techniques.

Biopsy↗
PubMed1995

Effect of intra-peritoneal fludarabine on rat spinal cord tolerance to fractionated irradiation.

The effect of fludarabine (9-beta-D-arabinosyl-2-fluoroadenine-5'- monophosphate), an adenine nucleoside analogue, on the tolerance of the spinal cord to fractionated irradiation was studied in a rat model. Anesthetized female Fisher 344 rats received irradiation to 2 cm of the cervical spine with a telecobalt unit (dose rate 1.14 Gy/min). Radiation was administered in two, four or eight fractions spread over a 48-h period with or without fludarabine. Animals assigned to combined therapy received two daily intraperitoneal injections of fludarabine (150 mg/kg) given 3 h prior to the first daily radiation fraction. It was found that fludarabine reduced the iso-effect dose required to induce leg paresis at 9 months after irradiation for all fractionation schedules. Dose modification factors of 1.23, 1.29 and greater than 1.27 were obtained for two, four and eight fractions, respectively. Fitting the data with the direct analysis method of Thames et al. with an incomplete repair model [18] showed that the potentiating effect of fludarabine may be mediated through reduction in the number of 'tissue-rescuing units' (InK). Alpha and beta values were slightly but not significantly decreased, whereas the alpha/beta ratio was unchanged. These features suggest that fludarabine did not significantly inhibit cellular repair processes but rather reduced the spinal cord tolerance by a fixed additive toxic effect on the same target cells. In rodent models, the combination of fludarabine and fractionated radiation has previously been found to yield a therapeutic gain, i.e., the drug enhanced tumor response to a greater extent than it reduced normal tissue tolerance.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

The role of fludarabine-induced apoptosis and cell cycle synchronization in enhanced murine tumor radiation response in vivo.

We have previously reported that fludarabine, an adenine nucleoside analogue, significantly enhances radiation-induced tumor regrowth delay and local cure in several mouse tumors. Although fludarabine potentiated tumor regrowth delay at various times from -36 h to +6 h in a SA-NH mouse sarcoma model, the greatest enhancement was observed when fludarabine was administered 24 h before irradiation. The purpose of this study was to understand the basis for in vivo enhancement of radiation efficacy by fludarabine. To examine the effect of fludarabine on DNA synthesis and cell cycle progression, tumor-bearing mice were given fludarabine by an i.p. route and then bromodeoxyuridine at various times up to 36 h, followed 0.5 h later by tumor harvest. Two-parameter flow cytometry analysis of the tumor cells using an anti-bromodeoxyuridine antibody demonstrated that an 800-mg/kg fludarabine dose stops DNA synthesis within 3 h with recovery starting at 12 h. By 24 h after fludarabine treatment, a synchronized wave of cycling tumor cells appeared in G2-M phase. The degree of DNA synthesis shutdown and the timing of the reinitiation of DNA synthesis and cell cycle progression were all fludarabine dose dependent. Interestingly, DNA synthesis reinitiated only at the G1-S boundary; cells in the S phase at the time of fludarabine administration appeared to disappear from the tumor population. To confirm these observations more directly, we pretreated tumor-bearing mice i.p. with chlorodeoxyuridine to mark the cells in the S phase, gave them fludarabine 0.5 h later, and then gave them iododeoxyuridine 0.5 h before tumor harvest. Flow cytometry analysis using antibodies specific for chlorodeoxyuridine- and iododeoxyuridined-labeled cells confirmed that cells in the S phase at the time of fludarabine administration never reinitiated DNA synthesis and disappeared from the tumor population. Immunohistological analysis of tumor sections obtained after fludarabine administration demonstrated that prelabeled S-phase cells took on an apoptotic appearance and gradually disappeared from the tumors. An in situ DNA end labeling assay demonstrated DNA fragmentation in these morphologically apoptotic cells. These results suggest that the mechanism of fludarabine enhancement of radiation response involves induced S-phase cell loss through an apoptotic pathway and subsequent synchronization of the remaining cells to a more radiosensitive cell cycle phase at the time of irradiation.

Animals↗
PubMed1994

Fludarabine improves the therapeutic ratio of radiotherapy in mouse tumors after single-dose irradiation.

PURPOSE: Fludarabine, an adenine nucleoside analogue, and an effective inhibitor of chromosome repair, was previously shown to synergistically enhance radiation-induced regrowth delay in three murine tumors. The purpose of this study was to assess whether fludarabine can increase the therapeutic ratio of radiotherapy in murine tumors, that is, to increase local tumor control without significantly modifying the radiation-induced normal tissue response. METHODS AND MATERIALS: Mice bearing 8-mm tumors in the right thigh (SA-NH sarcoma and MCA-K mammary carcinoma) were given 800 mg/kg fludarabine IP 3 h or 24 h before single doses of photon irradiation. Local tumor control was assessed by the TCD50 assay 100 days after treatment. Acute normal tissue toxicity was assessed in the skin (degree of epilation 30 days after irradiation) and in the jejunum (crypt regeneration assay), and late normal tissue toxicity was assessed by a leg contracture assay 120 days after treatment. RESULTS: In both tumors and with both drug schedules, fludarabine enhanced radiation-induced local tumor control (dose modification factors (DMF) of 1.24 (95% confidence limits 1.19-1.31) and 1.26 (95% confidence limits 1.20-1.32) for SA-NH, and 1.38 (95% confidence limits 1.25-1.50) and 1.35 (95% confidence limits 1.22-1.16) for MCA-K tumors). When given 3 h before radiation, fludarabine offered a slight protection from skin toxicity (DMF = 0.83, 95% confidence limits 0.77-0.86) but enhanced jejunum toxicity (DMF = 1.53). When fludarabine was given 24 h before irradiation, the reverse trend was observed (DMF = 1.11 (95% confidence limits 1.07-1.16) and 0.89, respectively). No enhancement of leg contracture was observed for either fludarabine schedule. CONCLUSION: The data presented here demonstrate that fludarabine can potentiate local tumor control induced by single-dose irradiation. While jejunum sensitization limited the relative effectiveness when fludarabine was administered 3 h before irradiation, a therapeutic ratio greater than one was always achieved when fludarabine was given 24 h before irradiation.

Animals↗
PubMed1994

Potentiation of radiation-induced regrowth delay in murine tumors by fludarabine.

Fludarabine (9-beta-D-arabinofuranosyl-2-fluoroadenine-5'-monophosphate), an adenine nucleoside analogue, has previously been shown to inhibit the repair of radiation-induced chromosome damage. Thus fludarabine may have therapeutic utility in combination with photon irradiation. The purpose of this study was to determine whether fludarabine could enhance radiation-induced murine tumor regrowth delay and to determine the most effective dose and schedule of the combination. A significant (P < 0.05) absolute regrowth delay enhancement was observed in three murine tumor models (SA-NH, a sarcoma; and MCA-K and MCA-4, mammary carcinomas) when fludarabine (800 mg/kg) was given 1 h prior to 25 Gy gamma-irradiation. While fludarabine enhanced radiation-induced tumor regrowth delay when given between -36 h and +6 h of radiation (SA-NH tumor), the greatest enhancement was observed when fludarabine was given at -24 h prior to irradiation (radiation dose modification factor of 1.82 at -24 h compared to 1.57 at -3 h prior to radiation). The degree of fludarabine enhancement (at -3 or -24 h) was dose dependent at doses above 200 mg/kg. When fludarabine and radiation were administered on a fractionated schedule (fludarabine given 3 h prior to radiation each day for 4 days), the dose modification factor increased to 2.14 (1.63 if the effect of fludarabine alone is subtracted). These results suggest that fludarabine enhances radiation-induced tumor regrowth delay in a more than additive fashion after both single and fractionated treatments, and the degree of enhancement is dependent on the sequence and timing of administration, the fludarabine dose, and the tumor type. Thus, fludarabine may have clinical potential as a radiation enhancer in the treatment of solid tumors.

Animals↗
PubMed1994

Endourologic management of obstructed hydrocalyx after blunt renal trauma.

Significant renal injuries caused by blunt trauma can produce early and delayed complications. We report a case of blunt trauma that resulted in what appeared to be a persistent encapsulated hematoma, but was in reality an obstructed hydrocalyx. Communication with the collecting system was re-established percutaneously by a method similar to that used in the management of caliceal diverticula.

Adolescent↗
PubMed1994

Transinguinal laparoscopy for nonpalpable testis.

Laparoscopy has changed the treatment approach in the patient with a nonpalpable testis. We report a novel method of laparoscopy to diagnose an atrophic, intra-abdominal testis through a contralateral inguinal peritoneotomy.

Child, Preschool↗
PubMed1993

Prospective comparison of in vitro normal cell radiosensitivity and normal tissue reactions in radiotherapy patients.

PURPOSE: This pilot study was undertaken to assess the relationship between in vitro radiosensitivity of different normal cell types and the type and severity of normal tissue reactions in individual patients after radiotherapy. METHODS AND MATERIALS: Twenty-one patients with head and neck cancer were studied prospectively; four with head and neck and two with breast cancer were studied retrospectively. The retrospective cases were chosen because they exhibited unusual (severe or minimal) normal tissue reactions after radiotherapy. Small skin biopsies and blood samples were obtained and used to generate in vitro fibroblast and lymphocyte cultures, respectively. Clonogenic assays were used to measure in vitro fibroblast and lymphocyte radiosensitivity after high- and low-dose rate irradiation. Head and neck patients were treated by conventional, hyperfractionated, or concomitant boost regimens, which have been found to yield an equal probability of late normal tissue reactions. The highest dose received by each normal tissue in the target volume was estimated using computed tomography treatment plans. The median patient follow-up time was 19 months (range: 13-25). RESULTS: The distributions of in vitro radiosensitivity parameters and the grade of tissue reaction scores in the patients showed a broad range between individuals. When in vitro parameters were compared to the acute and late tissue reactions, the radiosensitivity of fibroblasts, measured as surviving fraction at 2 Gy after high-dose rate irradiation, showed a highly significant correlation with the maximum grade of late effects (p < 0.0001 for the whole group and p = 0.0013 for the group of patients studied prospectively). No significant correlation was found between fibroblast radiosensitivity and maximum grade of acute effects or between lymphocyte radiosensitivity and either acute or late effects. CONCLUSION: We conclude that individuals vary in normal cell radiosensitivity, and that in vitro measurements of fibroblast radiosensitivity may predict the magnitude of late normal tissue reactions after radiotherapy. These preliminary results, however, need to be validated in a larger group of patients.

Biopsy↗
PubMed1993

Association of sickle cell disease, priapism, exchange transfusion and neurological events: ASPEN syndrome.

Priapism and acute neurological events are believed to be unrelated complications of sickle cell hemoglobinopathy. We describe a syndrome based on our experience and a review of the literature of significant neurological events after partial exchange transfusion to treat priapism in sicklemic patients. Severe headache is often the initiating symptom of this complex. The ensuing neurological events range from seizure activity to obtundation requiring ventilatory support. The proposed pathophysiology of these neurological events is related to cerebral ischemia after an acute increase in per cent total hemoglobin, concomitant decrease in per cent hemoglobin S and subsequent release of vasoactive substances during penile detumescence. We have termed this constellation of events the ASPEN syndrome, an eponym for association of sickle cell disease, priapism, exchange transfusion and neurological events. Early recognition and aggressive medical management resulted in complete reversal of neurological sequela.

Child↗
PubMed1993

Heterogeneity in the development of apoptosis in irradiated murine tumours of different histologies.

Fifteen different murine tumours were evaluated with respect to the degree of apoptosis development that occurs in the tumour tissue in the first few hours following irradiation in vivo. Animals were killed at 3 or 6 h following irradiation with 0, 2.5, 10 or 25 Gy. Apoptosis was scored as percent aberrant nuclei by microscopic examination of histological sections made from the tumour specimens. Results showed that three of four mammary adenocarcinomas, one ovarian adenocarcinoma, and one lymphoma displayed at least 10% apoptotic cells after 25 Gy, whereas five sarcomas, three squamous cell carcinomas, and a hepatocarcinoma did not. The time courses and dose responses were similar in those tumours that responded. These data were compared with the known response of these same tumours when analysed using conventional assays. The tumours that did respond by significant apoptosis had longer specific growth delays and lower TCD50 (dose to cure 50% of animals) doses, thus suggesting that an acute apoptotic response following irradiation may be a feature of certain tumours that respond well to irradiation. Additionally, this analysis revealed heterogeneity in the apoptotic response both within an individual tumour specimen and among different tumour types. These observations of intra and intertumour heterogeneity are consistent with the idea that the propensity for apoptosis in tumours is genetically regulated.

Adenocarcinoma↗
PubMed1993

Radiotherapy for head and neck neoplasms.

This review presents several relatively new areas of interest in radiotherapy for head and neck neoplasms. In the clinical arena, improving radiotherapy outcome by altering radiation schedules has undergone rigorous testing in patients with squamous cell carcinomas. The initial results show that such an approach increases the local control rate in subsets of patients by 10% to 15%. Randomized multi-institutional studies, sponsored by large cooperative groups, are ongoing to assess the relative merits of various altered fractionation strategies. In the field of laboratory investigations, attempts have been made to develop tests reliable in predicting the radiation response of tumors and normal tissues. If successful, it would be possible to design the optimal radiation regimen for individual patients based on the results of these tests, thereby improving the therapeutic ratio. Technologic innovations have enhanced the flexibility to conform radiation to the target volume so that higher doses can be delivered to the tumor without exceeding the tolerance of the surrounding normal tissues. Large-scale clinical studies evaluating the value of conformal therapy are being developed. Finally, long-term results of radiotherapy in the management of chemodectomas have been reported in recent years. These data have clearly defined the role of radiotherapy in the treatment of skull-base paragangliomas.

Clinical Trials as Topic↗
PubMed1993

Latex allergy and anaphylaxis.

Latex allergy is a potential cause of unexplained intraoperative anaphylaxis. Latex allergic reactions range from mild wheal and flare episodes to cardiovascular collapse. High-risk patient populations merit latex allergy testing. Preoperative prophylactic regimens may prevent or attenuate serious intraoperative latex allergic reactions in susceptible individuals. Additional work is needed to better characterize and understand latex allergy.

Anaphylaxis↗
PubMed1992

Intrinsic radiosensitivity of normal human fibroblasts and lymphocytes after high- and low-dose-rate irradiation.

The existence of heritable radiosensitivity syndromes and clinical observations in radiotherapy patients suggests that human cellular radiosensitivity differs among individuals. We report here an in vitro study of radiosensitivity in 30 fibroblast and 29 lymphocyte cultures obtained from cancer patients and controls. In 25 cases, both fibroblasts and lymphocytes were obtained from the same donors. Fibroblasts were cultured from skin biopsy samples, and peripheral T-cell lymphocytes were cultured from blood. Clonogenic survival assays were performed by using high- and low-dose-rate irradiation; lymphocytes were in G0 phase and fibroblasts in confluent plateau phase. Various end points were calculated and compared (i.e., surviving fraction at 2 Gy, initial slope of the survival curve, and doses resulting in 10 and 1% survival, respectively). Depending on the end point, the coefficient of variation of the survival parameters ranged from 31 to 68% for lymphocytes and 21 to 41% for fibroblasts following high-dose-rate irradiation. Similar ranges were obtained after low-dose-rate irradiation. Variance analysis performed on replicate assays in cultures derived from the same patient showed that variation due to technical or sampling errors was significantly lower than variation between individuals (P = 0.00034 and 0.014 for fibroblasts and lymphocytes, respectively). No correlation was observed between the radiosensitivity of lymphocyte and fibroblast cultures derived from the same donors. We conclude that there is significant variation in normal cell radiosensitivity among individuals. On the other hand, comparisons of lymphocyte and fibroblast radiosensitivities suggest that tissue-specific characteristics, such as differentiation status, may variably modulate radiosensitivity.

Breast Neoplasms↗