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Biomedical subjects

G R Prout

Publications and source records attributed to G R Prout.

At least 73 records · Page 4Linked to original sources

Effect of 514.5-nm argon ion laser radiation on hematoporphyrin derivative-treated bladder tumor cells in vitro and in vivo.

The therapeutic effect of hematoporphyrin derivative (HpD) plus 514.5-nm argon ion laser radiation was compared to HpD plus 630-nm argon ion laser-pumped dye laser radiation in experimental urinary bladder transitional cell carcinoma models. Cultured human bladder cancer cells (EJ) containing HpD were 2.8-fold more sensitive to 514.5-nm radiation than to 630-nm radiation as measured by clonogenic capacity. The relative effectiveness of 514.5-nm versus 630-nm light was approximately proportional to the spectral absorbance for cell-bound HpD at these wavelengths. HpD-sensitized photoirradiation was studied in solid tumors produced by a) the subcutaneous inoculation of cells from murine bladder tumors induced by N-[4-(5-nitro-2-furyl)-2-thiazoyl]formamide (CAS: 24554-26-5) into female C3H mice (MBT-2 tumor) and b) the intravesical instillation of N-methyl-N-nitrosourea (CAS: 684-93-5) into the urinary bladders of female Wistar rats. The tumors were exposed to 144 J/cm2 laser light 24-48 hours following ip injection of 20 mg HpD/kg body weight. By 24-48 hours, animals that received HpD and light of either wavelength had partially or completely necrosed tumors. Control groups showed no necrotic changes. Regression of MBT-2 tumors was also investigated. Seven of 14 and 6 of 12 animals had nonpalpable tumors 1 week after treatment with 514.5-nm and 630-nm light, respectively. Tumors in control groups demonstrated no regression. Spectral transmittance from 630 nm to 514.5 nm decreased by about 4% for 130- to 160-micron-thick sections of canine urothelium and bladder submucosa-muscularis. The results of this study indicate that HpD plus 514.5-nm laser radiation may be an effective treatment for small or superficial malignant lesions of the urinary bladder.

Animals↗

Production and characterization of mouse monoclonal antibodies to human bladder tumor-associated antigens.

Monoclonal antibodies (McAbs) to human bladder carcinoma were generated by fusion of NS-1 mouse myeloma cells with spleen cells from BALB/c mice immunized with either cultured human bladder cancer cells or cells obtained from a fresh surgically removed bladder tumor. Four hybridomas which reacted strongly with bladder tumor cells and not to normal skin fibroblasts or urothelial cells were identified and cloned by limiting dilution to obtain monoclonality. One McAb, 3G2-C6, raised with cultured tumor bladder cells MGH-U1 (EJ) as the immunogen reacted more strongly to the bladder tumor lines tested than any of the other McAbs resulting from various fusion experiments. Hybridoma 3G2-C6 was found to secrete murine immunoglobulin G1 and to produce high titer ascites fluid when grown in BALB/c mice. Results from quantitative enzyme-linked immunosorbent assays on a panel of more than 35 cell lines demonstrated that McAb 3G2-C6 reacted with several bladder tumor cell lines 50 to 90 times more than with normal transitional urothelium. Two kidney and two testicular tumor lines also bound 10 times more 3G2-C6 than with normal cells. The 3G2-C6 antigen was only marginally detected on a number of other cancer and noncancerous cells tested such as breast and lung tumor cells, melanoma, fetal cells, and peripheral blood lymphocytes. To identify the antigen 125I-labeled membrane components from MGH-U1 cells were extracted with detergent, immunoprecipitated with Protein-A bound 3G2-C6, and analyzed by sodium dodecyl sulfate-gel electrophoresis. This revealed that McAb 3G2-C6 binds to a Mr 90,000 cell surface component. Indirect immunofluorescence microscopy with fluorescein isothiocyanate-anti-mouse immunoglobulin G also identified the antigen on the surface of cultured and fresh tumor cells and detected the antigen on 16 of 17 Grade 3 bladder tumor specimens as well as on some kidney and testicular tumor cells. This study confirms the potential of the hybridoma technique for producing McAbs capable of identifying tumor associated antigens which may be useful in the diagnosis and treatment of bladder cancer.

Animals↗

Establishment and characterization of four human bladder tumor cell lines and sublines with different degrees of malignancy.

We have established four human bladder tumor cultures, designated MGH-U1 to -U4 (also known as EJ, HM, RN, and RB in some previous reports). All have been grown in culture for over 30 passages and were free of Mycoplasma contamination. Characterizations of these cell lines were performed. These include isozyme profile, morphology with light and scanning electron microscopes, karyotype, growth rate, DNA content by flow cytometry, presence of cell surface ABH isoantigens, tumorigenicity in nude mice, lactic acid dehydrogenase isozymes, and colony formation in soft agar. Results obtained from these characterizations confirm that MGH-U1 and -U2 are sublines of a previously established bladder tumor cell line, T-24. These results also show that MGH-U3 and -U4, derived respectively from a grade 1 tumor and an urothelium biopsy with severe atypia, are likely to be independent human bladder cell lines and different from other transitional cell bladder carcinoma cell lines reported. The study further demonstrates that these four cell lines/sublines have different degrees of malignancy and a close correlation, in biological and malignant characteristics, between the cells in culture and those in the original tumors. Therefore these cultures may represent cells at different stages of malignant progression. These can be useful models for studies of the development and progression of bladder tumors and detection and treatment of bladder tumors of different grades and stages.

ABO Blood-Group System↗

Mitomycin C intravesical therapy in noninvasive bladder cancer after failure on thiotepa.

Mitomycin C 40 mg in 40 ml water was administered intravesically every week for 8 consecutive weeks to 60 patients with superficial bladder cancer. All patients had failed treatment with intravesical thiotepa and had evaluable disease. An objective response of 50% or greater reduction in measured tumor mucosal involvement was obtained in 68% of patients. Forty-two percent of the patients achieved a complete response, and this included 50% of patients with Grade III disease and 70% of patients with a T1 tumor. Median response duration in complete responders was 12.2 months with a range of 3.5 to 24.3 + months. Fifty-five percent of patients are still responding. Therapy was generally well tolerated, and in contrast to thiotepa, myelosuppression was not the dose-limiting effect. One third of all patients experienced symptoms of local irritation, and skin reactions were seen in 12% of patients.

Adult↗

Cisplatin and full dose irradiation for patients with invasive bladder carcinoma: a preliminary report of tolerance and local response.

Twenty-seven patients with invasive bladder carcinoma (clinical stages T2 to T4) who were not candidates for cystectomy were treated by transurethral resection, cis-diamminedichloroplatinum (cisplatin) and full dose radiotherapy according to protocol 8 of the National Bladder Cancer Collaborative Group A. Nausea and vomiting occurred in 74 per cent of the patients but were mild in 41 per cent. Maximum followup was 27 months and during that time 3 significant toxic reactions occurred: renal failure, systemic sepsis and a transient partial small bowel obstruction. Of 17 evaluable patients complete responses of the primary bladder cancer to the treatment were achieved in 11 of 13 with stages cT2 and cT3 cancer and in 2 of 4 with stage cT4 disease. The members of National Bladder Cancer Collaborative Group A have found transurethral resection, cisplatin and full dose external beam radiotherapy practical clinically. Longer followup will be necessary to determine if the observed high initial complete response rate of the tumor indicates real lasting benefit for these patients.

Acute Kidney Injury↗

Identification of vessels by lectin-immunoperoxidase staining of endothelium: possible applications in urogenital malignancies.

Ulex europaeus agglutinin I binds specifically to vascular endothelium of various human tissues irrespective of the blood group type or secretive status of the tissue. Using this property, we have developed an immunoperoxidase technique to identify vessel structures in urogenital tissues. Most vessels in malignant and nonmalignant tissues of bladder, prostate and testis of different blood types (A, B, AB and O) can be readily identified by this method. This technique has potential application in detecting tumor cell invasion of vessels and in studying distribution of vessels in relation to various normal and pathological events.

ABO Blood-Group System↗

Cystoscopic fluorescence detector for photodetection of bladder carcinoma with hematoporphyrin derivative.

Hematoporphyrin derivative, a fluorogenic substance, tends to accumulate at higher levels in most tumor tissues after intravenous injection. These properties provide a potential application for the detection of malignant tumors. We report the development and in vitro evaluation of an instrument designed to excite and detect low levels of hematoporphyrin derivative fluorescence emitted by tumor cells in the bladder after hematoporphyrin derivative administration. The light source of the instrument for specific hematoporphyrin derivative excitation is at a wavelength of 405 nanometers. Optical fibers for both the excitation and detection of fluorescence were bundled in a single 7 French ureteral catheter which can be inserted into a standard cystoscope. The fluorescence is initially detected as an electrical signal which is amplified and then converted into an audio signal. The interference of the cystoscope illuminating white light to the fluorescence signal is eliminated by a phase-sensitive electronic circuit. The instrument thus allows simultaneous viewing of the bladder and detection of hematoporphyrin derivative fluorescence produced from tumor tissues. In vitro testings using hematoporphyrin derivative solutions on filter paper indicated that the instrument detects as little as 30 nanograms per milliliter of hematoporphyrin derivative and has relatively low noise levels from normal tissues. The utility of this instrument for hematoporphyrin derivative photodetection of small tumors and carcinoma in situ in bladder is currently under clinical evaluation.

Animals↗

Malignant teratoma of the testis with an isochromosome no. 12, i(12p), as the sole structural cytogenetic abnormality.

Cytogenetic analysis of a malignant teratoma of the testis revealed a hypotriploid karyotype with the presence of an isochromosome of the short arm of chromosome 12, i(12p), as the only structural abnormality. Most cells contained 3 copies of chromosome 12 and 2 copies of the i(12p) isochromosome. Thus, the genetic material located on the short arm of chromosome 12 was multiplied in the teratoma cells, being represented by 7 copies compared to 2 to 3 copies of other chromosomes. This finding confirms the reports that the i(12p) isochromosome is a specific chromosomal abnormality in cases of malignant testicular tumors.

Adult↗

Correlation of vascular invasion and metastasis in germ cell tumors of testis--a preliminary report.

A preliminary study was conducted to correlate vascular invasion with metastasis in testicular tumors. A lectin-immunoperoxidase procedure was used to identify the vessels. Together with standard hematoxylin and eosin staining, the presence of tumor cells in vessels was detected. Primary tumor specimens from 37 cases and retroperitoneal lymph nodes from 10 cases of germ cell tumors were studied. The results showed good correlations between vascular invasion in primary tumors and metastasis to lymph nodes in nonseminomatous germ cell tumors, and between vascular invasion in retroperitoneal lymph nodes and the presence of other metastases. Eight of the 9 nonseminomatous germ cell tumor patients with vascular invasion had metastases, and all 4 patients that had vascular invasion in the lymph nodes had other metastases. However, there seemed to be no correlation between vascular invasion and clinical staging in cases with seminoma in which retroperitoneal lymph node dissection was not performed. These observations, although based on a limited number of cases, warrant more extensive investigations.

Blood Vessels↗

Visualization of urothelial blood group isoantigens A and B using direct biotin-labeled antibodies and avidin-biotin-peroxidase complex.

The loss of blood group isoantigens from the surface of bladder tumor cells has been correlated with the potential invasiveness of the tumor. Development of simple and reliable methods for detection of these isoantigens should facilitate the general clinical use of this test for predicting malignant potential in low grade, low stage cancer of the bladder. We now report a direct peroxidase technique for the detection of isoantigens A and B by utilizing the specific interaction between biotin and avidin, and the capability of labeling a single antibody with multiple biotin molecules. Antibodies specific to the isoantigens A and B were purified from human antisera by affinity chromatography using an immunoabsorbent containing chemically synthesized antigenic determinants. The purified antibodies were directly labeled with biotin. An avidin-biotin-peroxidase complex was used to bind the biotinylated antibody for the peroxidase staining reaction of the isoantigens on tissue section. Application of this technique to formalin-fixed, paraffin-embedded bladder tissue and tumor sections yielded specific and strong stainings of the isoantigens with low background staining. The potential clinical application of this method requires further evaluation.

ABO Blood-Group System↗

Testicular tumors: delay in diagnosis and influence on survival.

Of 133 men with testicular abnormalities, 52 underwent orchiectomy within a month of symptoms, 41 delayed seeking medical advice and 40 were treated initially for conditions other than testicular cancer. Death occurred in 20 of the 89 patients with nonseminomatous germ cell tumors; in 16 of these, diagnosis had been delayed. Prompt diagnosis and treatment could reduce mortality from these neoplasms to less than 10 percent.

Adolescent↗

Nonrandom chromosomal changes in transitional cell carcinoma of the bladder.

Nine cases of transitional cell carcinoma (eight from the urinary bladder and one from the ureter; six noninvasive and three invasive) were subjected to detailed cytogenetic analysis with a G-banding method. The synchronization of primary cultures with methotrexate for high-resolution banding was performed in five cases. In the remaining four cases, the chromosomes were obtained from short-term cultures after prolonged (16 hr) exposure to Colcemid. Two cases were near-tetraploid, one was hypotriploid, and six were near-diploid (three hyperdiploid and three hypodiploid). All but one case showed various structural abnormalities in the karyotype. The chromosomal changes ranged from the presence of only two abnormal chromosomes (markers) to complex karyotypes with as many as 15 markers. In most tumors, the origin of the marker chromosomes could be readily deciphered. The nonrandom chromosomal aberrations included: (a) an isochrosome of the short arm of chromosome 5 (three cases); (b) monosomy of chromosome 9 found in four cases (this was the sole abnormality in one case); (c) involvement of chromosome 8 as an isochromosome of the long arm (two cases) or loss of the short arm due to deletion (one case) or translocation (one case); and (d) interstitial deletion of chromosome 13 (three cases). Our results indicate that the formation of i(5p) and monosomy 9 may be the primary karyotypic changes in two subgroups of transitional cell carcinoma. Involvement of chromosomes 8 and 13, on the other hand, seems to be a result of secondary karyotypic evolution. Two invasive tumors showed the presence of secondary clones, with additional structural chromosome aberrations superimposed on those already existing in the main cell population. In both cases, the additional aberrations involved the short arm of chromosome 11, resulting in loss of genetic material from the short arm. The short arm of chromosome 11, is the putative site of an oncogene which has been isolated from human bladder carcinoma cell lines. Deletion of the 11p was also seen in one case of noninvasive transitional cell carcinoma localized in the ureter; the material from 11p was probably translocated to chromosome 13. These findings suggest that the loss of genetic material from the short arm of chromosome 11 is a secondary event in the karyotypic evolution of transitional cell carcinoma, probably related to the invasive behavior of the tumor.

Aged↗