Goblet cell carcinoid of appendix: ultrastructural features and histogenetic aspects.

Light and electron microscopic examination of a goblet cell carcinoid revealed cells with pleomorphic neurosecretory-type granules, cells containing mucin some of which also contained these granules and less differentiated cells lacking the aforementioned features. Recent embryologic and anatomic studies of developing avian and mammalian gut, respectively, show that intestinal APUD cells are probably of endodermal origin. Therefore, mixed carcinoid tumors such as the goblet cell variant could arise in crypt base stem cells.


Plexiform tumors of the uterus: ultrastructural study.

Two cases of plexiform tumor of the uterus are presented. Both were incidental microscopic findings in leiomyomatous uteri and had the typical branching cords of small, polygonal cells with scanty cytoplasm and vesicular nuclei. Ultrastructurally the cells had features of smooth muscle differentiation such as actin-like filaments, dense bodies, peripheral dense plaques, pinocytotic vesicles, and incomplete basal lamina. Their close relationship to capillaries resembled the structure of glomus tumor and vascular leiomyoma and suggested histogenetic kinship among these neoplasms.


Amylase-producing ovarian neoplasm with pseudo-Meigs' syndrome and elevated pleural fluid amylase: case report and ultrastructure.

Elevated amylase activity was noted in the pleural effusion of a patient who was later found to have a stage I low-grade serous papillary ovarian neoplasm. The effusion resolved spontaneously after resection of the ovarian tumor, which contained large amounts of amylase activity. The ultrastructure of the tumor epithelium resembled that of normal salpinx. Secretory-type cells were present with apical, variably electron-dense secretory granules and cytoplasmic glycogen deposits. Amylase activity in the ovarian neoplasm probably resulted from the presence of functioning endosalpingeal-type epithelium in the tumor. In cases of effusion with unexplained amylase elevations, the possibility of serous ovarian neoplasia should be considered, even in the absence of demonstrable extra-ovarian dissemination.


Gallium nitrate (NSC-15200) induced toxicity in the rat: a pharmacologic, histopathologic and microanalytical investigation.

Administration of gallium nitrate to rats resulted in the formation of renal precipitates which occluded tubular lumina. When analyzed with a combination of scanning electron microscopy and x-ray energy spectrometry, these precipitates were found to contain gallium complexed with calcium and phosphate. Injection of gallium nitrate also resulted in hypercalciuria, although serum calcium levels remained unaltered. Administration of an osmotic diuretic, isosorbide, prior to gallium treatment resulted in the formation of fewer renal precipitates and histopathologic changes than in the nondiuresed animals. Diuresis did not alter gallium serum pharmacokinetics, the 24 hour cumulative renal excretion of gallium or the extent of the drug-induced hypercalciuria. However, isosorbide pretreatment significantly reduced the urinary concentrations of both gallium and calcium. The data presented indicate that diuresis reduces the severity of gallium-induced renal lithiasis and subsequent renal accumulation of gallium by diluting the urinary concentration of gallium and calcium thereby lowering the incidence of interaction of these two elements within the kidney tubule.


Angioimmunoblastic lymphadenopathy with retinitis and drug related exacerbations: a clinicopathological case study.

A patient with angioimmunoblastic lymphadenopathy with dysproteinemia was followed over a three year period from diagnosis to death. He presented with arthralgias, uveitis and respiratory insufficiency and developed hyperuricemic renal failure upon institution of treatment. Aggressive combination chemotherapy was required to reverse progressive thrombocytopenia and pulmonary involvement. A complete remission was achieved twice. There was a striking temporal relationship between the administration of antibiotics or allopurinol and exacerbations of the disease. Hypocomplementemia and transient evidence of vasculitis suggested the presence of immunecomplexes. Serial lymph node biopsies showed the progression of this disorder from a pleomorphic immunoblastic proliferation to a lymphocyte-depleted, fibrotic process, in parallel with a decline from hyper- to hypogammaglobulinemia. This case illustrates the broad clinical spectrum of angioimmunoblastic lymphadenopathy with dysproteinemia and suggests that aggressive treatment is necessary in selected patients.


Effect of metastatic cancer on platelet monoamine oxidase activity and serotonin metabolism.

Platelet monoamine oxidase (MAO) activity and urinary excretion of 5-hydroxyindoleacetic acid (5-HIAA), serotonin, tyramine and tryptamine were measured in patients with a variety of metastatic cancers and in healthy control subjects. The patients with cancer had higher platelet MAO activity and lower urinary excretion of 5-HIAA, sertonin and tyramine than the healthy subjects. The urinary tryptamine excretion was not significantly different in the two groups. At the present time it is unclear if the alterations in platelet MAO activity and monoamine excretion are casually related.


Medullary carcinoma of the thyroid with atypical patterns.

A case of a highly invasive thyroid carcinoma, which occurred in a 68-year-old woman, was studied by light and electron microscopy, and histochemical and biochemical analysis. Light microscopical, histochemical, and biochemical features were consistent with a diagnosis of a calcitonin-producing, amyloid-rich medullary carcinoma; electron microscopical patterns, mainly the presence of lumina, microvilli, and extensively dilated cytoplasmic E.R., were reminiscent of the ultrastructural features of the follicular carcinoma. Electron-dense bodies interpreted as "secretory granules" were very scarce. This case appears very similar to the case recently presented by Valenta et al. and interpreted as a microfollicular carcinoma. The atypical features of our case of MCT seem to indicate that this tumor should be included in a group of atypical MCT; these should be kept separate from the typical (differentiated) ones on morphological, functional, and prognostic grounds.


Fibroadenoma in oral contraceptive users: a histopathologic evaluation of epithelial atypia.

We have reviewed histologically a series of 120 fibroadenomas which formed part of the case material from a previous case-control epidemiological investigation of the relationship between oral contraceptive use and breast disease. We evaluated the epithelial component of the fibroadenoma for degree of cytologic atypia. This study indicates that the reduced risk for fibroadenoma among long-term users of oral contraceptives does not vary according to the degree of epithelial atypia present. This is in contrast to our previously reported findings for fibrocystic disease, in which the decreased frequency of occurrence of the disease in long-term users of oral contraceptives was found only for cases with no minimal epithelial atypia.


Incidence and histopathology of metastases of mammary carcinoma in biopsies from the posterior iliac crest.

Bone marrow biopsies were taken from the posterior iliac crest in 532 women with unilateral breast cancer. Metastatic tumors were found in 10% of the biopsies. In a group with negative radiological examinations of the skeleton, the incidence of positive bone marrow biopsies was 1.6%. In a group with radiologically detectable metastases in the skeleton 28% of the biopsies were positive. In the latter group 43 out of 45 individuals with positive biopsies had negative x-rays of the pelvis. Histopathologically, 19% of the metastatic tumors were osteolytic, 65% were osteoblastic and 16% did not influence the bone structure. The fibrous reaction in and around the bone marrow tumors was similar to that found in the primary tumor. In 74% the morphological pattern was consistent throughout the biopsy, whereas in 26% the morphology was different in different parts of the biopsy. No specific histopathology was observed in the individuals with negative radiological examinations of the skeleton. Bone marrow biopsy of the posterior iliac crest does not seem to be helpful as a routine method in the initial staging of mammary carcinoma but may contribute to establish the degree of tumor spread in individuals with positive or suspicious x-ray of the skeleton.


Estradiol and testosterone levels in the peripheral and ovarian circulations in patients with endometrial cancer.

Estradiol and testosterone levels in the peripheral and ovarian circulation of patients with endometrial cancer and of postmenopausal women were studied. These levels proved to be the same in the two groups. The ovaries of neither group secreted estradiol. The testosterone level in the ovarian circulation of all examined women was four times higher than that in the peripheral circulation, thus indicating the ovarian origin of testosterone.


Pleomorphic xanthoastrocytoma: a distinctive meningocerebral glioma of young subjects with relatively favorable prognosis. A study of 12 cases.

Twelve cases of a distinctive form of supratentorial astrocytoma occurring in young subjects (ages 7 to 25) are reported. The tumors were superficial and involved the leptomeninges extensively. The tumor cells display marked pleomorphism, including bizarre giant cells and a number of mitotic figures, but no necrosis. Many contain large amounts of lipid in their cytoplasm and are surrounded by reticulin fibers, thus simulating a mesenchymal tumor. For these reasons, some examples of this tumor have been previously interpreted to represent meningocerebral fibrous xanthomas. Immunoperoxidase technique performed in nine of the twelve cases has, however, established the presence of glial fibrillary acidic protein in the tumor cells, which are therefore considered to be astrocytic. By electron microscopy many tumor cells are surrounded by basal laminae, accounting for the abundant reticulin network demonstrable in silver preparations. Since subpial astrocytes are known to be partly covered by a basal lamina, it is likely that they are the cells of origin for this neoplasm. In contrast to its pleomorphic cytology, the biological behavior of this tumor appears to be relatively favorable, and long survival times (up to 25 years) have been recorded in some cases. (These tumors are distinct from intracranial fibrous xanthomas of mesenchymal derivation. Cells of the latter are negative on GFAP stain.)


Endocrine profile in a patient with familial breast cancer: a case-control study.

Urinary and blood hormonal profiles were studied throughout a monthly cycle in a patient with familial breast cancer. Two comparison cohorts (one high-risk and one low-risk) were studied concurrently. Findings disclosed that our breast cancer-affected patient showed a distinctive hormonal pattern characterized by significant elevation throughout the cycle of plasma estrone, estradiol, and prolactin. Save for a depression in plasma FSH in the early follicular phase, this hormone, as wells as LH and progesterone patterns in our patient, were similar to the comparison cohorts. Urinary estrone and estradiol patterns in our patient were elevated early in the follicular phase. Our patient also showed a depression in urinary estrone, estradiol, and estriol following ovulation, which persisted throughout the luteal phase. Blood and urinary hormone patterns in the high-risk cohort were not demonstrably different from the low-risk cohort, with the exception of plasma prolactin. The results on the latter hormone showed an unexpected significant depression throughout most of the menstrual cycle in this low-risk cohort. We conclude that estrone and estradiol elevations, as clearly evidenced in our breast-cancer-affected patient, may provide clues that ultimately might be used as an etiologic discriminant for breast cancer risk and which may also play a pathogenic role in this disease. Since this involved a single patient, our conclusions must be interpreted cautiously.


Genital cancer in wives of penile cancer patients.

We identified the wives or ex-wives of 227 males of the 256 reported with cancer of the penis to the New York State Cancer Registry from Upstate New York from 1960-64. Utilizing the Registry, death certificates, hospital and physician records, we ascertained those wives who developed cancer at any site from 1951-1975. We generated expected numbers of cases of cancer at each site by applying the age specific incidence rates experienced by women of a specific age in a specific year designated by the age of the wife of the index case in each year, estimating withdrawals from age-specific death rates. Thus, our expected numbers are based on the experience of the women in Upstate New York with traits like those of spouses of the men in the same population with cancer of the penis. We found significantly more cases of cancer of the cervix than expected. This was not true for other sites of cancer.


Cancer of the genital tract among the Ibo women in Nigeria.

An analysis of female genital tract cancer among the Ibos of Nigeria has proven to be preponderantly cervical cancer. It is therefore obvious that the practice of ritual circumcision among the Ibo men does not protect the women from cervical cancer. The most important etiological factors contributing to cervical cancer among the Ibo women are early marriage, early age of coitus, and grand multiparity. Corpus cancer is relatively rare among the Ibo women.


Diagnostic value of fiberoptic bronchoscopy in lung cancer presenting as mediastinal mass(es).

The diagnostic yields of prebonchoscopy sputum specimens and fiberoptic bronchoscopy (including brushings, washings, and/or biopsies) were determined in 35 patients who presented primarily with middle or anterior mediastinal and/or paratracheal mass(es) on chest radiographs. The diagnosis was confirmed on histopathology of tissue obtained by needle biopsy, mediastinoscopy, thoracotomy, and/or autopsy. Thirty-one of the patients were found to have primary bronchogenic carcinomas. At bronchoscopy, extrinsic compression of trachea and/or bronchi was visualized in 23 (Group I) and the tracheobronchial tree appeared normal in 12 (Group II). Prebronchoscopy sputa gave a positive yield in only three of the 35 (8%) patients; the yield was similar in both groups of patients. One or more of the bronchoscopic modalities were diagnostic in 69% of Group I patients but were not helpful in Group II patients. The diagnostic yield of brushings, washings, and biopsies in Group I patients was 52%, 61%, and 37%, respectively. The addition of biopsies to washings and/or brushings did not significantly alter the yield. Washings and brushings are recommended as useful, non-invasive procedures in diagnosing middle mediastinal masses with extrinsic compression. Sputum cytology gives a very low yield.


Pulmonary resection for metastatic nonosteogenic sarcoma.

Analysis was made of 112 patients who underwent resection of metastatic pulmonary noldules for nonosteogenic sarcoma at the Mayo Clinic from 1950--76. The 5-year post-thoracotomy survival was 29% with a median survival of 18 months. There has been no substantial change between 1950 and 1976 in post-thoracotomy survival among all patients who received this operation here. Prognosis was especially grim (p less than 0.01) for patients with pre-thoracotomy disease-free intervals of less than 12 months, and for those experiencing extrathoracic recurrences vs. further pulmonary recurrent tumor (p = .01) following thoracotomy. The following discriminants were not significantly associated with postthoracotomy survival: age, sex, histology, number of lesions fully excised, or site of lung lesions. We conclude that the therapeutic value of thoracotomy in the treatment of metastatic nonosteogenic sarcomas has not changed here during the past quarter of a century. Whether the recent more common use of thoracotomy in metastatic disease of this type is therapeutically worthwhile cannot be clearly determined from the data available.


The natural history of breast cancer patients with brain metastases.

One hundred one breast cancer patients with brain metastases (BM) were reviewed. The median survival from BM was 4.0 months. Seventy percent were receiving chemotherapy at the diagnosis of BM and 43% were showing a clinical response. Prolonged survival was seen in patients who underwent surgical resection, those with the brain as site of first metastasis, and those with a long free interval who survived the initial 5 months after BM. Long-term survivors (greater than 18 months from BM) demonstrated indolent disease by all parameters measured.


Acute nonlymphocytic leukemia and acute myeloproliferative syndrome following radiation therapy for non-Hodgkin's lymphoma and chronic lymphocytic leukemia: clinical studies.

Seven cases of acute nonlymphocytic leukemia (ANLL) and one case of a malignant myeloproliferative syndrome have been seen after extensive radiation therapy for non-Hodgkin's lymphoma or chronic lymphocytic leukemia. A myeloproliferative syndrome with abnormalities in granulocytic, erythrocytic, and thrombocytic cell lines was present in all patients and in seven patients preceded ANLL by 2--18 months. The median time to the development of ANLL after primary disease therapy was 61 months (33--98 range). The leukemia was extremely refractory to therapy and median survival after diagnosis of ANLL was two months (range 0--9 months). Leukemia was seen only in those patients who received multiple courses and multiple techniques of radiation therapy.


DNA content in human uterine cervical dysplasias.

The DNA content of nuclei extracted from separate biopsies of normal cervical epithelium and from dysplastic sites in the same patient was determined biochemically. Mean percentage increase in the DNA content of dysplasias (mild: 143.3 +/- 14.9; moderate: 225.8 +/- 51.1; and severe: 359.8 +/- 45.8) was found to be statistically significant (F-Test: p less than 0.001) over the normal values in control cervical tissues.


Enhancement of thermal killing by polyamines. IV. Effects of heat and spermine on protein synthesis and ornithine decarboxylase activity.

Protein synthesis is shown to be very heat-sensitive in Chinese hamster cells. It is shut off completely following 15-20 min at 42 degrees C whereas RNA and DNA syntheses are affected only after much longer exposure times. Cells recover from inhibition of protein synthesis upon transfer to 37 degrees C. The degree of recovery is inversely related to the duration of heat exposure and it fits cell survival quantitatively. Cells which become temporarily heat-resistant by prior heat-treatment, are able to recover translational capacity even after a very long exposure to heat (4 h at 42 degrees C). Spermine, which enhances heat-induced cell killing, does not increase the response to heat of protein, RNA and DNA synthesis. Ornithine decarboxylase (ODC, EC 4.1.1.17) activity is lost exponentially following a 20 min lag period during exposure at 42 degrees C. The half-life observed (12 min) is in agreement with the reported values of half-life of decay of ODC in other systems. It is concluded that the loss of activity is due to the shut-off of translation. The activity of ODC is recovered upon transfer to 37 degrees C. The presence of spermine during heating does not affect the loss of enzyme activity but delays its recovery by about 3 h upon transfer to 37 degrees C.


Shapes of carcinogenic benz[a]anthracenes: the crystal and molecular structure of 1-methylbenz[a]anthracene.

The crystal structure of 1-methylbenz[a]lanthracene, which is weakly carcinogenic, has been determined by application of direct methods to single-crystal X-ray diffractometric data and refined by least squares to R = 0.09 over 845 independent reflections. Crystals are monoclinic, space group P2(1), with a = 8.491(2), b = 7.138(2), c = 10.500(2)ABEta  = 95.06(01), Z = 2. As in other benz[a]anthracenes, the K-region bond C(5)-C(6) is short [1.34(1)A]. The distinctive bay geometry, with a methyl group opposite to a hydrogen, H(12), peri to another hydrogen, H(11), has a long bond C(13)--C(18) = 1.47(1)A in the bay, and the angular benz-ring is inclined at 16.5 degrees to the mean plane of the anthracene fragment. The methyl carbon atom is 0.79 A out of the mean molecular plane (or 0.19 A out of the plane of the benz-ring) and the 1.50 A long C(1)-methyl bond makes angles of 117 degrees and 125 degrees at C(1).


Isolation of a DNA polymerase alpha-associated regulatory protein from calf thymus.

A regulatory protein for DNA polymerase alpha, responsive to noncomplementary deoxyribonucleoside triphosphates, has been isolated from calf thymus. The regulatory protein was separated from DNA polymerase alpha using Affi-Gel Blue and gel filtration. The regulatory protein had a molecular weight of approximately 70,000 as determined by gel filtration, and its activity was nondialyzable, heat labile, and abolished by pronase treatment. In the presence of regulatory protein, DNA polymerase alpha activity, measured by using polydeoxyadenylate-oligodeoxythymidylate as template primer, was inhibited by 2'-deoxyguanosine 5'-triphosphate in a parabolic-competitive fashion [Ki = 15 +/- 1 (S.E.) microM] and by 2'-deoxycytidine 5'-triphosphate in a linear-competitive manner (Ki = 162 +/- 23 microM). Neither the four natural ribonucleoside triphosphates nor 2'-deoxyadenosine 5'-triphosphate inhibited the DNA polymerase-regulatory protein system to any significant extent. The regulatory protein by itself had no effect on either DNA polymerase alpha activity or the Km for template primer. These results indicate that deoxyribonucleoside triphosphate pools may be involved in the regulation of cellular DNA synthesis through a direct effect on DNA polymerization.


Response of 9L tumor cells in vitro to spirohydantoin mustard.

The effects of spirohydantoin mustard (SHM), a potential antitumor agent for central nervous system tumors, in the in vitro 9L rat brain tumor model were studied. In cell culture medium at 37 degrees, the drug was totally detoxified within 30 min. Dose-response curves for exponentially growing and plateau-phase cells were similar and indicated that a small fraction of cells were resistant to SHM. When exponentially growing cells were treated with SHM (5 microgram/ml for 1 hr), recovery from potentially lethal damage occurred within 28 hr. When cells were perturbed by SHM, the S, G2, and M phases were prolonged, there was a G2 block, some cells entered mitosis, but few divided, and cells tended to accumulate in mid-S, then moved synchronously to G2-M. The rate at which cells moved was concentration dependent and was much slower at high concentrations. The ability of SHM to both synchronize cells and block DNA synthesis may be useful in multiagent therapy regimens.


In vitro cytokinetic response of human colon cancer cells to cis-dichlorodiammineplatinum(II).

The cytokinetic response of a human colon carcinoma cell line to cis-dichlorodiammineplatinum(II) was investigated using flow cytometry of DNA content, autoradiography after pulse and continuous tritiated thymidine exposure, and mitotic accumulation after continuous Colcemid treatment. With increasing concentration and exposure time, cis-dichlorodiammineplatinum(II) delayed and then blocked cycle traverse in S and G2 phases. After prolonged treatment with high concentrations of cis-dichlorodiammineplatinum(II), an additional block in G1 or at the G1-S boundary was established. Irreversibility of cell cycle distribution changes after prolonged observation periods suggests cell death in G2, S, and G1 compartments.


Cellular accumulation and disposition of aclacinomycin A.

The cellular accumulation and disposition of the anthracycline antitumor antibiotic aclacinomycin A (ACM) were compared to those of daunorubicin. Although both drugs were avidly accumulated by cells, intracellular concentrations of ACM were two to three times those of daunorubicin. Whereas lowered temperature (0 degrees) reduced intracellular accumulation of both drugs, 10 mM sodium azide had no effect on accumulation of either ACM or daunorubicin. Both drugs exited from cells placed in drug-free medium, a process that was reduced at 0 degrees but not altered by 10 mM sodium azide. Unlike whole cells, isolated nuclei accumulated more daunorubicin than ACM. This process was not altered at 0 degrees. Both drugs were lost from nuclei placed in drug-free buffer, a process that was reduced at 0 degrees. Unlike daunorubicin, which localized in cell nuclei, ACM localized in the cytoplasm with no detectable nuclear fluorescence. Although both drugs produced dose-dependent inhibitions of [3H]thymidine and [3H]uridine incorporation by L1210 and P388 cells, ACM inhibited both processes at lower concentrations than did daunorubicin. While daunorubicin inhibited [3H]thymidine incorporation more effectively than [3H]uridine incorporation, the reverse was observed with ACM.


In vivo cell cycle synchronization of the murine sarcoma 180 tumor following alternating periods of hydroxyurea blockade and release.

The durations of the cell cycle intervals of the murine Sarcoma 180 tumor were determined by computer analysis of the fraction-labeled mitosis curve following tritiated thymidine administration. This tumor has a usual total cell cycle duration of 19.6 hr, a DNA synthetic time of 8.3 hr, and a growth fraction of 1. Approximately 38% of cells are in S phase at one time. Hydroxyurea (HU) infusions (i.v.) at 1.17 mg/hr into tumor-bearing mice rapidly inhibit tumor DNA synthesis. Following a 5-hr HU infusion, 58% of all tumor cells are in S phase, and maximal tumor mitotic rates after release of the HU blockade are double control rates. HU was infused for 5 hr, followed by 7 hr of Ringer's solution, and then another 5 hr of HU. Following this 2-cycle blockade, 70% of tumor cells are in S phase, predominantly in early S phase and at the G1-S junction. After release, peak mitotic rates are 2.5 times control. The duration of the intermitotic time of the tumor following HU infusion is less than the total cell cycle time of control tumor. Cycles of HU infusion and release, timed according to the predetermined duration of the cell cycle intervals, will synchronize significant increments of S phase or mitotic cells of the Sarcoma 180 tumor during predictable periods of time.


Failure of rotenone to interfere with 17 beta-estradiol action in the rat uterus.

The involvement of rotenone in rat mammary carcinogenesis has been suggested to occur through estrogenic effects. This hypothesis was tested by determining the extent of rotenone inhibition of 17 beta-estradiol binding to the estrogen receptor and of the 17 beta-estradiol-induced uterotrophic response in ovariectomized Sprague-Dawley rats. Estradiol binding to the uterine estrogen receptor in the presence of rotenone was determined by charcoal assay and Scatchard analysis. Additionally, 17 beta-estradiol-receptor interactions were assessed on sucrose density gradients. No inhibition of binding was observed in either assay with ratios of rotenone/17 beta-estradiol in excess of 10,000. Finally, an in vivo approach was used to extend the in vitro data. Silastic capsules containing rotenone or 17 beta-estradiol were implanted in various combinations into eight groups of ovariectomized Sprague-Dawley rats (four rats/group). After five days, uteri were removed and weighed. An analysis of variance revealed that rotenone neither interfered with 17 beta-estradiol-induced uterine weight gain nor displayed any uterotrophic properties by itself. Results from these three procedures demonstrate that rotenone does not act as an estrogen or as an estrogen antagonist. Additionally, there were no other effects attributable to rotenone.


Comparison of four new cell lines from patients with adenocarcinoma of the ovary.

Permanent human tumor cell lines COLO 110, COLO 316, COLO 319, and COLO 330 were established from four patients with serous cystadenocarcinoma of the ovary. COLO 110 was derived from primary tumor tissue; COLO 316, COLO 319, and COLO 330 were derived from cells in malignant effusions. COLO 110 and COLO 316 grew as monolayers of epithelioid cells in culture; COLO 319 and COLO 330 grew as vermiform, floating colonies of epithelioid cells in culture. Epithelial-like morphology was confirmed by transmission electron microscopy. All four cell lines had marker chromosomes and double minute chromosomes. Giemsa banding revealed chromosomes 1, 3, 6, and 7 were involved in markers in all four lines, and chromosomes 2, 4, 5, 9, 11, and 15 were involved in markers in three of the cell lines. Marker chromosomes with possible homogeneous staining regions were observed in COLO 319. Estrone was elaborated by three of the lines, but neither chorionic gonadotropin, carcinoembryonic antigen, nor estrogen or progesterone receptor proteins were detected. Each cell line demonstrated a distinctive isozyme phenotype. These cell lines are maintained in active culture and in a cell bank for distribution to other investigators.


Effects of combination drug therapy on the subcutaneous and pulmonary growth of a slow and a fast-growing C3H/He mammary carcinoma.

Changes in susceptibility to treatment with Cytoxan, methotrexate, 5-fluorouracil, and Adriamycin, single or in combination, have been studied during the initial and progressive stages of s.c. and pulmonary (via tail vein injection) growth of two transplanted syngeneic C3H/He mammary carcinomas. One tumor was fast growing, reaching a size of 3 mm from a 1-mm s.c. implant in 7 days; the second tumor would grow to the same size in 30 days. The tumor with the slower growth rate was more susceptible to drug treatment, manifested by delayed growth as well as by prevented growth. The slower-growing tumor also remained susceptible longer, when treatment was delayed, than did the faster-growing tumor. Pulmonary growth was more often prevented by drug treatment than was s.c. growth. Tumor implants s.c. which had reached palpable size could be reduced temporarily to impalpable size by effective drug treatment but were rarely cured. The importance of early treatment relative to the time of tumor implantation was indicated when early treatment with a single drug proved more effective than did delayed treatment with a more potent combination of drugs.


Ornithine decarboxylase activity and DNA synthesis after treatment of cells in culture with 12-O-tetradecanoylphorbol-13-acetate.

The ability of the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) to induce the enzyme ornithine decarboxylase (ODC) and to stimulate DNA synthesis was studied in four different cell types in vitro. The effects of this agent on each cell type were different: (a) in hamster embryo cells, TPA induced ODC but had no effect on DNA synthesis; (b) TPA induced ODC and stimulated DNA synthesis in BALB/c 3T3 mouse cells; (c) it did not induce ODC in human fibroblasts but did stimulate DNA synthesis; and (d) it induced neither ODC nor DNA synthesis in rat embryo fibroblasts. In contrast to the effects of TPA, ODC was induced and DNA synthesis was stimulated in all cell types by fresh serum-containing medium. Treatment of the cells with a combination of fresh medium and TPA resulted in an approximate summation of the effects of treatment with each agent alone. These results emphasize the differences in the responses of various cells to TPA. They also show that in some cells, at least, the induction of ODC and stimulation of DNA synthesis following TPA treatment can be regulated independently.


Tumorigenicity and intracisternal A-particle expression of hybrids between murine myeloma and lymphocytes.

Hybrids of BALB/c lymphocytes and a murine myeloma, a tumor that expresses intracisternal A-particles, were obtained with polyethylene glycol as the fusogen. The karyotype, tumorigenicity, and A-particle expression of the hybrid clones were assessed. All the hybrid clones analyzed were tumorigenic and expressed intracisternal A-particles even when they were the result of a fusion event between two lymphocytes and one myeloma cell in which no loss of chromosomes was detected. The tumors that developed following inoculation of hybrid cells into BALB/c mice (1 x 10(6) cells/mouse) were karyotypically identical to the inoculated cells. It appears that the two myeloma cell phenotypic traits analyzed (tumorigenicity and A-particle expression) are dominant.


Induction of hepatic neoplastic lesions in mice with a single dose of hycanthone methanesulfonate after partial hepatectomy.

Experiments were designed to determine whether hycanthone methanesulfonate (1-([2-(diethylamino)ethyl]amino)-4-(hydroxymethyl)thioxanthen-9-one monomethanesulfonate), an antischistosomal drug, and its analog, IA-4-N-oxide (8-chloro-2-[2-(diethylamino)ethyl]-2H-[1]benzothiopyrano[4,3,2-cd]indazole 5-methanol monomethanesulfonate), will induce neoplastic lesions in the livers of mice not infected with Schistosoma mansoni. All the mice received a single i.m. injection of hycanthone methanesulfonate (76 mg/kg), IA-4-N-oxide (80 mg/kg), or an equivalent volume of the solvent, 0.9% NaCl solution, 42 hr after partial hepatectomy. Of the mice receiving hycanthone methanesulfonate and living 200 days or longer, hepatocellular carcinoma was seen in 11.5% and liver sarcoma was seen in 4.2%. This type of malignant neoplasm was not seen in the animals receiving either IA-4-N-oxide or 0.9% NaCl solution. In addition, mice receiving hycanthone methanesulfonate showed a significantly higher incidence of both type 1 (43% compared to 21% in controls) and type 2 (21% compared to 12% in controls) hepatocyte neoplasms. Mice receiving IA-4-N-oxide showed no increased incidence of neoplasms.


Risk factors for nodular sclerosis and other types of Hodgkin's disease.

The age-adjusted incidence rates of nodular sclerosis in Los Angeles County from 1972 to 1975 were 58% lower in Mexican-Americans and 42% lower in blacks than in other whites. They were the same for each sex among other whites and the curve of age-specific incidence peaked in young adulthood. The incidence of nodular sclerosis was directly associated with social class. In contrast, the rates for other histological varieties of Hodgkin's disease (lymphocyte predominance, mixed cellularity, lymphocyte depletion) were only 12% lower in Mexican-Americans but 34% lower in blacks. In other whites, the rates were 92% higher in men and increased gradually with age while there were no clear trends with social class. These characteristics support the hypothesis that, at least for purposes of etiology, the nodular sclerosis form of this disease should be considered a distinct entity. Self-administered questionnaire were completed by 218 of the 1972 to 1973 Hodgkin's disease patients and 218 individually matched neighbourhood controls. Significantly high risk ratios for Hodgkin's disease were found for prior appendectomy (risk ratios = 1.9, p = 0.01) and for past amphetamine use (risk ratios = 3.0, p = 0.01). The elevated risk associated with amphetamine use had been found in a previous study.


Effects of local tumor hyperthermia on the growth of solid mouse tumors.

The sensitivity to local tumor hyperthermia (43 degrees, 60 min) of a spectrum of eight different solid mouse tumors (Lewis lung carcinoma, M5076 ovarian carcinoma, colon carcinoma 38, colon carcinoma 26, mammary adenocarcinoma C3HBA, mammary adenocarcinoma 16C, glioma 26, and B16 melanoma) was investigated. A microwave (2.45-GHz) apparatus produced localized heating of the tumors without generation of whole-body hyperthermia. The temperature at the center of the heated tumors was regulated to within +/- 0.1 degrees while the temperature uniformity within the tumor was +/- 0.5 degrees. The local hyperthermia treatments reduced the size and retarded the growth of the treated tumors compared with control values for each of the tumors tested. The faster-growing Lewis lung carcinoma and B16 melanoma were the least responsive to treatment, while the slower-growing colon 38 and M5076 ovarian carcinomas were the most responsive. Multiple treatments resulted in longer grwoth delays and greater tumor growth inhibition than did single treatments. No consistent difference in life span between the control and treated groups was measured, and only five of 188 treated animals were cured.


Catecholestrogen synthesis and metabolism by human breast tumors in vitro.

The activities of enzymes which synthesize and metabolize catecholestrogens were studied in biopsy samples of human breast neoplasms. Estrogen 2-hydroxylase, a cytochrome P-450-dependent enzyme, was present in both benign and malignant neoplasms but not in normal breast tissue. Catechol O-methyltransferase activity was present in all samples examined and was significantly higher in malignant tumors [549 +/- 31 (S.E.) pmol/20 min/mg protein] than in benign neoplasms (226 +/- 41 pmol/20 min/mg protein) or in normal breast tissue (133 +/- 28 pmol/20 min/mg protein). There was no correlation, however, between estrogen 2-hydroxylase and catechol O-methyltransferase activities. The enzymes responsible for the synthesis and metabolism of catecholestrogens are present in some breast tumor specimens, suggesting that in such tissues these metabolites may be formed in vivo.


Specific inhibitors directed at the individual components of ribonucleotide reductase as an approach to combination chemotherapy.

It had been shown previously that the ribonucleotide reductase from mouse tumor consisted of two nonidentical components (Tris and dye fractions, each prepared from the 20 to 40% (NH/)2SO4 protein fraction containing the ribonucleotide reductase activity by blue dextran-Sepharose chromatography). The individual components either separated or present in the intact enzyme can be specifically and independently inhibited by different compounds. The Tris fraction component was inhibited by 4-methyl-5-amino-1-formylisoquinoline thiosemicarbazone while the dye fraction component was inactivated by pyridoxal phosphate:BH4- and the dialdehyde derivative of inosine (Inox) and 5'-deoxyinosine prepared by the periodate oxidation of inosine and 5'-deoxyinosine. The intact enzyme could be completely inhibited by any of these compounds. Reductase activity was restored by reconstitution with the exogenous components. The individual components of the reductase in the intact Ehrlich tumor cell could also be specifically inhibited. Activity in the crude cell-free extracts prepared from 4-methyl-5-amino-1-formylisoquinoline thiosemicarbazone- or Inox-treated cells was restored by the addition of the appropriate exogenous component. These data suggest that combinations of inhibitors of ribonucleotide reductase which specifically inhibit the components may be useful in the treatment of cancer.


Thermosensitization by sulfhydryl compounds of exponentially growing Chinese hamster cells.

The effect of various sulfhydryl compounds on the survival of exponentially growing monolayer cultures of Chinese hamster cells (HA1) heated to temperatures of 37-43 degrees was examined. Concentrations of cysteamine which were nontoxic or minimally toxic at room temperature or 37 degrees became increasingly toxic at elevated temperatures, greatly potentiating the killing produced by heat alone in the absence of cysteamine. This enhancement of hyperthermia-induced cell killing increased with increasing cysteamine concentration, increasing duration of cysteamine exposure, and increasing temperature. Studies with synchronized Chinese hamster cells heated at 43 degrees for 1 hr in the presence of 16 mM cysteamine demonstrated that the potentiation of heat killing occurred in all phases of the cell cycle. Similarly, enhancement of hyperthermia-induced cell killing was seen for asynchronous cells exposed to 2-amino-ethylisothiourium bromides and cysteine, but the magnitude of the effect differed for the various sulfhydryl compounds.


Ontogeny of the rat hepatic receptor for 2,3,7,8-tetrachlorodibenzo-p-dioxin and its endocrine indepence.

The ontogeny of the 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) receptor was studied in Sprague-Dawley rats by quantitation of the receptor in liver cytosol using isoelectric focusing in polyacrylamide gel. No differences by sex in the receptor concentration were seen at any of the ages studied. Newborn, 21-day-old, and 42-day-old rats contained significantly more receptor in the liver cytosol than did 56-day-old rats. There was no significant difference in the receptor concentration in liver cytosol from 7-day-old rats compared to that from 56-day-old rats. The maximum receptor concentration was found in cytosol from 21-day-old rats [36.1 +/- 24.0 (S.D.) fmol/mg protein]. Adult rats (56 days old) contained the lowest concentration of receptor (13.3 +/- 6.3 fmol/mg protein). The level of TCDD receptor in liver cytosol from adult rats was not significantly changed by orchiectomy, ovariectomy, adrenalectomy, or hypophysectomy. The maximum for TCDD receptor concentration at puberty corresponds to the reported maximum for the induction of aryl hydrocarbon hydroxylase activity. However, no further conclusion can as yet be drawn concerning the regulation of the TCDD receptor.


Protein synthesis in liver tissue under the influence of a methylcholanthrene-induced sarcoma in mice.

Amino acids are incorporated at increased rates into hepatic proteins in tumor-bearing humans and animals. In this study, we hoped to elucidate whether this is an expression of increased hepatic protein synthesis or altered isotope dilution in the precursor pool(s). Liver tissue from sarcoma-bearing mice (MCG 101) showed increased specific activities of arginine and leucine in hepatic proteins after i.p. injection of these precursors. The specific radioactivity of leucine in the free amino acid incorporation rate into proteins was also found in incubated liver slices and in a cell-free system of incubated free and membrane-attached polysomes. The increased amino acid incorporation was the net result of increased as well as decreased relative turnover of various hepatic proteins. The hepatic content of RNA was increased, but hepatic DNA and protein content was unchanged in tumor-influenced livers. Increased amino acid incorporation into hepatic proteins in tumor-bearing animals and also probably in cancer patients is due to a net increased hepatic protein synthesis, probably not confined to acute-phase reactants only.


Disturbance of phospholipid metabolism during the selective destruction of tumor cells induced by alkyl-lysophospholipids.

Alkyl-lysophospholipids inhibit the growth of Meth A sarcoma cells in vitro. In contrast, murine bone marrow macrophages are not sensitive to the destructive effect of these substances. Since alkyl-lysophospholipids are antimetabolites in the synthesis of 3-sn-phosphatidylcholine, tumor cell destruction can be correlated with the disturbance of this metabolism. A decreased synthesis of 3-sn-phosphatidylcholine is accompanied by an increased degradation of cellular 3-sn-phosphatidylcholine in the presence of alkyl-lysophospholipids. As a consequence, endogeneously formed lysophospholipid accumulates, although the lysophospholipase is found to be stimulated. This accumulation of endogeneous lysophospholipids might be due to the fact that a high percentage of these compounds contain an alkyl bond which cannot be split by a lysophospholipase. On the other hand, the reacylation of the formed lysophospholipids is partially blocked as the lysophosphatidylcholine acyltransferase is inhibited by the added alkyllysophospholipids. An accumulation of potentially cytotoxic lysophospholipids in tumor cells might be an additional factor in the tumor cell destruction by alkyl-lysophospholipids.


Association of macrophage activation with antitumor effect on rat syngeneic fibrosarcoma by Nocardia rubra cell wall skeleton.

The antitumor activities of the cell wall skeleton (CWS) of Nocardia rubra were demonstrated for syngeneic fibrosarcoma (AMC-60) in ACI/N rats in regard to macrophage activation. In the 24-hr cytolytic test, activated macrophages which were fractionated from peritoneal exudate cells induced by i.p. injection of Nocardia CWS showed significant cytolytic activity for [125I]iododeoxyuridine-labeled tumor cells. Activated macrophages also strongly inhibited [3H]thymidine incorporation into the tumor cells during the 24-hr cytostatic test. When tumor cells were inoculated s.c. with activated macrophages in the Winn-type transfer assay, subsequent tumor growth was significantly inhibited. Repeated i.p. injection of the CWS seemed to enhance these antitumor activities of macrophages. The therapeutic effect of Nocardia CWS was assessed with the ascites tumor and with the solid tumor inoculated i.m. into the hind leg. In the former treatment, repeated i.p. injections completely prevented the accumulation of ascites fluid and resulted in prolongation of the survival period. The peritoneal macrophages harvested from these survivors had a strong cytolytic activity for tumor cells in the cytolytic test. In the latter treatment, repeated intratumoral injections inhibited the growth of primary tumor and prevented metastasis. Furthermore, peritoneal resident macrophages from these tumor-bearing rats treated intratumorally with the CWS were found to be cytolytic for tumor cells in the cytolytic test.


Development of androgen-independent spindle cell tumors from androgen-dependent medullary Shionogi carcinoma 115 in androgen-depleted nude mice.

When Shionogi carcinoma 115 (SC115, undifferentiated medullary carcinoma showing compact cell pattern and containing androgen receptor) was transplanted into male and female DS mice, it grew only in males. In contrast to this strict androgen dependency in DS hosts, tumors composed of spindle-shaped cells appeared in more than 80% of cases when SC115 tumor was inoculated into female or castrated male nude athymic (BALB/c-nu/nu) recipients. These spindle cell tumors neither contained cytosol androgen receptor nor showed biologically defined androgen dependency. As spindle cell tumors could be serially transplanted in DS mice but not in BALB/c-+/+ mice and as the original SC115 (medullary carcinoma showing a compact cell pattern) tumor and the spindle cell tumor had many identical chromosome abnormalities, these two types of tumors seem to have a common origin in spite of their morphological, biochemical, and biological differences. Since spindle cells could not be detected histologically in SC115 tumors maintained in intact male DS mice, the present results seem to suggest that SC115 cells may change their morphological, biochemical, and biological characteristics within one passage in androgen-depleted nude athymic mice.


Enhancement of interferon antitumor action by sodium butyrate.

Sodium butyrate together with interferon enhances the antitumor effect of interferon in vivo. When Sarcoma 180 TG cells are inoculated in mice, the mean survival time and final survival rate are greatly increased compared to those for treatment with interferon alone. Similarly, a significant delay in the mean survival time is observed when mice inoculated with L1210 cells are treated with sodium butyrate and interferon. This effect could be due at least in part to a potentiation of interferon action on the tumor cells.


Characterization of liver-type alkaline phosphatase from human gastric carcinoma cells (KMK-2) in vitro.

Alkaline phosphatase was extracted from human gastric carcinoma cells (KMK-2) under long-term culture, and its biochemical and biological properties were investigated. The enzyme was extremely heat labile and was inhibited significantly by L-homoarginine, but only slightly by L-phenylalanine, so that it was classified as a liver-type alkaline phosphatase. Comparative studies with liver and early placental alkaline phosphatases revealed that the enzymes all showed a similar extent of inhibition by amino acids, heat stability, immunological character, molecular, and other biochemical properties. However, KMK-2 alkaline phosphatase was more similar to early placental enzyme in electrophoretic and gel filtration pattern. This liver-type alkaline phosphatase was found ultrastructurally on microvilli of KMK-2 cells, but not on the lateral structurally on microvilli of KMK-2 cells, but not on the lateral surface with interdigitating folds. Prednisolone markedly decreased the content of the present isozyme. Although the present phenotype was stable during long-term culture in regard to the isozyme properties, the original cancer cells from which the cell line had been derived were L-phenylalanine sensitive and moderately L-homoarginine sensitive. This indicated that phenotypic change occurred on cultivation of cancer cells in vitro.


Junctional intercellular communication pattern of cultured human breast cancer cells.

Junctional intercellular communication between several established human breast cancer cell lines and a variety of mammalian cells has been examined. All the cancer cell lines were found to be either noncommunicators or nonselective communicators. This contrasts with normal human mammary epithelium which shows selectivity in junctional communication. Loss of selectivity in junctional communication appears to be a general feature of cultured human breast cancer cells.


Correlation between glucocorticoid receptor and cytolytic response of murine lymphoid cell lines.

By fusion between murine thymoma lines which are either homozygous (r+/r+ or r-/r-) or hemizygous (r+/r-) for the glucocorticoid receptor structural gene r, hybrid clones have been obtained which carry one, two three, or four copies of the r+ allele. These hybrids contain different amounts of normal glucocorticoid receptor as the result of a r+ gene dosage effect. Measurements of the cytolytic response of these hybrids to dexamethasone indicate a tight correlation between receptor content and sensitivity.


Increased production of mutagenic metabolites of carcinogens by tissues from senescent rodents.

Activation of the procarcinogens benzo(a)pyrene and 2-fluorenamine by liver homogenates (S9) prepared from senescent male CFN rats and C57BL/6J mice resulted in an enhanced production of mutagenic metabolites when compared to young rodents, as indicated by an enhancement of the induced reversion frequency in a Salmonella typhimurium bioassay. Similar results were observed when carcinogen activation was mediated by purified hepatic microsomes, indicating that the age-related differences in carcinogen activation did not result from aging changes in carcinogen metabolism involving non-microsomal mechanisms. The metabolites of many procarcinogens are thought to be the ultimate carcinogens in mammals. Therefore, the present findings are consistent with the hypothesis that some fraction of the markedly increased of neoplasia observed in senescent mammals is a result of age-related alterations in the metabolism of chemical carcinogens.


Pharmacokinetic studies during phase I trials of high-dose thymidine infusions.

Thymidine infusions (75 g/sq m/24 hr) were administered to 12 cancer patients as part of a Phase I study. Thymidine and thymine measurements, by high-pressure liquid chromatography, were made on plasma and urine from eight of these patients. Only the pharmacokinetic aspects of these studies are reported in this paper. Millimolar thymidine and thymine concentrations were achieved in all patients and maintained for 120 hr during each of three courses of infusion. The half-life of thymidine was approximately 100 min following cessation of infusion. The half-life of thymine was much longer but could not be accurately determined because it did not decline as a first-order rate function. The cerebrospinal fluid:plasma ratios at steady state for thymidine and thymine were 0.29 and 1.03, respectively. Total body clearance of thymidine ranged from 95 to 266 ml/min/sq m, and 41 to 67% was by kidney clearance of intact thymidine. Calculations and comparison to other studies at lower infusion rates (micromolar plasma thymidine) indicate that thymidine is metabolized significantly by organs in addition to the liver and that, at millimolar plasma thymidine, total body metabolic processes of thymidine are saturated as is the secretory portion of kidney clearance.


Inhibition of DNA synthesis in cultured lymphocytes and tumor cells by extracts of betel nut, tobacco, and miang leaf, plant substances associated with cancer of the ororespiratory epithelium.

The high incidence of oropharyngeal, esophageal, and laryngeal cancers in certain parts of the world has been ascribed to conjugated tannins found in certain folk medicinal herbs. We extracted miang leaf and betel nut with phosphate-buffered saline (0.14 M NaCl, 0.15 M potassium phosphate buffer, pH 7.4) and found that the extracts inhibited [3H]thymidine incorporation by phytohemagglutinin-stimulated human lymphocytes and by rat mammary tumor and mouse L-cells in logarithmic growth. Pretreating the lymphocytes for 1 or 4 hr with the extracts inhibited phytohemagglutinin-induced thymidine incorporation 72 hr later. At concentrations of 2.5 volumes % or lower, miang and betel nut extracts inhibited thymidine incorporation by 40 to 98% without any apparent signs of toxicity as demonstrated by the 66Rb equilibrium assay. In addition, neither extract inhibited cytotoxicity of rat mammary tumor cells by immune syngeneic spleen cells. The molecular weights of the inhibitory factors were between 1,000 and 10,000 daltons as determined by ultrafiltration and were unaffected by boiling for 3 min or by treatment with alcohol and, therefore, are probably not proteins. This in vitro demonstration of inhibition of DNA synthesis by these plant extracts presumably enriched for conjugated tannins may relate to inhibition of growth of rats and chicks fed conjugated tanin-contaminated sorghum feed. The carcinogenic potential of either these extracts or conjugated tannins is not yet established.


Comparative antitumor effects of Corynebacterium parvum, Bordetella pertussis, Bacillus Calmette-Guérin, and levamisole alone or in combination with cyclophosphamide in the CaD2 murine mammary adenocarcinoma system.

The antitumor efficacy of various immune stimulants [Corynebacterium parvum, Bordetella pertussis, and Bacillus Calmette-Guérin (BCG)] and levamisole alone or in conjunction with cyclophosphamide (CY) was studied in the CaD2 mammary adenocarcinoma system using schedules developed previously with C. parvum and CY. Weekly systemic treatment with C. parvum, B. pertussis, or BCG was effective in controlling tumor growth and had equivalent antitumor effects, but weekly treatment (or a single treatment) with levamisole was ineffective. Weekly treatment with B. pertussis was better than treatment given only once, but repeated treatment with C. parvum or BCG was not more effective than a single treatment with these agents. When administered as a single systemic injection, C. parvum was superior to B. pertussis in controlling tumor growth, but a single systemic injection of BCG was as effective as C. parvum. Systemic administration of immune stimulants had variable effects on survival, which were sometimes not correlated with effects on tumor size. Combined treatment with BCG and CY was significantly more effective than CY treatment alone in controlling tumor growth in most trials, as was combination treatment with C. parvum and CY. Combination treatment with B. pertussis and CY was not better in prolonging survival than CY alone. Levamisole rarely improved the antitumor effect of CY chemotherapy and had no effect on survival compared to the effect of CY alone.


Effect of 5-fluorouracil on RNA metabolism in Novikoff hepatoma cells.

The resistance of a Novikoff hepatoma cell line (N1-S1/FdUrd) to 5-fluorouracil (FUra) was reversed by the inclusion of inosine in the culture medium. As the concentration of inosine in the medium was increased, there was a marked increase in the uptake of [14C]FUra and conversion to nucleotides with a corresponding increase in the incorporation into RNA. While FUra alone had no effect on this resistant cell line, the combination of FUra plus inosine altered the levels of ribose 1-phosphate but not 5-phosphoribosyl 1-pyrophosphate, altered the maturation of precursor ribosomal RNA by blocking the formation of 18S RNA, altered the methylation of the ribosomal RNA, and caused inhibition of the growth of these cells. No evidence was obtained that fluorodeoxyuridine 5'-monophosphate was formed in the N1-S1/FdUrd cells as a result of treatment with FUra plus inosine. In addition, the metabolism of [3H]deoxycytidine in the presence of FUra plus inosine in the intact N1-S1/FdUrd cells did not indicate significant inhibition of thymidylate synthetase as evidenced by the levels of deoxyuridine 5'-monophosphate or conversion to thymidine 5'-monophosphate.


Cell lines from human colon carcinoma with unusual cell products, double minutes, and homogeneously staining regions.

Two human colon carcinoma cell lines derived from the same tumor specimen were characterized. The cell lines, COLO 320 and COLO 321, have amine precursor uptake and decarboxylation cell properties, such as ectopic production of norepinephrine, epinephrine, serotonin, adrenocorticotropic hormone, and parathyroid hormone. The cells were morphologically different from most colon cell lines. Double minutes (DM) were initially present in nearly 100% of the metaphases. In a few subcultures of COLO 320, DM have persisted for 1.5 years. However, in COLO 321 and some subcultures of COLO 320, a loss of DM was observed and new marker chromosomes with homogeneously staining regions were observed. These unusual cell lines should be valuable for studies of apudomas of the colon and the cytogenetic phenomena of DM and homogeneously staining regions.


Promotion of azoxymethane-induced intestinal cancer by high-fat diet in rats.

Promotional properties of a high-fat diet in intestinal cancer were studied by feeding a 30% beef fat diet to 8 groups of rats (25 rats/group) for time periods varying from 1 to 21 weeks after 8 weekly s.c. injections of azoxymethane (AOM) (8 mg/ kg). Two other groups were fed the high-fat diet, one for 8 weeks prior to and the other during AOM injections. A 5% fat diet was fed to rats when not on the 30% fat diet and to a control group of 25 animals. High fat diet increased intestinal tumor frequency up to 2-fold when given for at least 4 weeks after but not during or prior to AOM injections; this increase occurred even after a prolonged interval (10 weeks) between the last AOM injection and the high-fat diet. In general, tumor frequency increased according to the length of time animals were fed the high-fat diet after AOM. Therefore, the high-fat diet in this model exhibited most of the properties of promoters developed from murine skin cancer, thus adding support to the concept that excess dietary fat acts at the promotional phase of carcinogenesis.


Adaptation of Sinclair swine melanoma cells to long-term growth in vitro.

Sinclair swine melanoma usually regresses in vivo. In the present study, swine melanoma cells were adapted to long-term growth in culture. The morphology of cultured melanoma cells ranged from dendritic to cuboidal, similar to that described for human melanoma cells. Doubling times of the swine melanoma cells were also similar to those of human melanoma cells in vitro. 3,4-Dihydroxy-L-phenylalanine oxidase-positive cells were detected by light microscopy, and melanin and premelanosomes were detected by electron microscopy. Cell cultures could be propagated from progressing, partially regressed, and primary cutaneous lesions, as well as from visceral metastases. Thus, it appears that, under these cell culture conditions, Sinclair swine melanoma cells can be adapted to prolonged growth in vitro.


Effect of hypoxia on 67Ga incorporation into human granulocytes.

Human granulocytes accumulate 67Ga when incubated under anoxic conditions and exclude the isotope when oxygenated. When incorporated, 67Ga is associated with the lysosomal fraction of the granulocytes. The ability of granulocytes to exclude the isotope except under hypoxic conditions may explain 67Ga localization in abscesses and some tumor masses.


Effects of caffeine on purine metabolism and ultraviolet light-induced lethality in cultured mammalian cells.

Caffeine, at doses which enhance the killing action of ultraviolet light, inhibits both de novo synthesis and the utilization of exogenous purines in cultured CHO-K1, a Chinese hamster ovary cell line. The decrease in synthesis was measured as inhibition by caffeine of the accumulation of phosphoribosylformylglycineamide or of phosphoribosylaminoimidazolecarboxamide, the fourth and ninth intermediates, respectively, in the de novo biosynthetic pathway. The effect is dose dependent, with a caffeine concentration of 7.5 mM producing a 90% reduction in 15 min. Interference with utilization of exogenous purines was seen as a substantial decrease in the conversion of [14C]hypoxanthine, [14C]adenine, or [14C]guanine into their respective di- and triphosphates in the presence of caffeine. Purine deprivation either by starvation of purine-requiring mutants or by treatment of parental cells with methylmercaptopurine ribonucleoside, a known inhibitor of purine synthesis, results in a partial sensitization to killing by ultraviolet light which can be maximized by the addition of caffeine. Thus, one of the ways by which antimetabolites and caffeine act to enhance ultraviolet light killing may be by interference with the supply of purine nucleotides needed for repair.


Kidney tumors induced in rats by the antischistosomal drug niridazole.

An increased incidence of kidney tumors was found in MRC rats fed the antischistosomal drug niridazole at four dose levels in the diet. Histologically, the adenomas and adenocarcinomas were solid papillary, clear cell, and tubular types, with the latter type predominating. Seven mesenchymal tumors were found among the 107 renal epithelial neoplasms. Severe nephrosclerosis occurred in both treated and control rats and has been suggested as important in renal carcinogenesis. Niridazole is considered a potent inducer of epithelial kidney tumors.


Effects of hormones on growth and alpha-lactalbumin activity in the transplantable rat mammary tumor MCCLX.

We have examined the growth and differentiation properties of the transplantable mammary tumor line of the A X C rat, MCCLX. The MCCLX tumor grows rapidly and exhibits high activity of the milk protein alpha-lactalbumin (alpha LA) (40% of the level in midlactating mammary gland) during serial passage in estrogen-supplemented (E+) intact males. Hypophysectomy of hosts bearing established tumors results in rapid tumor regression and rapid loss of alpha LA protein activity. Treatment with ovine prolactin three times daily (total of 2 mg/day) completely restores alpha LA activity and results in a partial restoration of tumor growth. In females, MCCLX grows only in E+ virgins or in pregnant hosts. During lactation, tumor growth ceases. However, alpha LA activity is equally high in E+, pregnant, and lactating females. Behavior of MCCLX apparently depends on circulating lactogenic hormones. When hormone levels are very high, as in E+ intact males or pregnant females, both growth and alpha LA activity are stimulated. In lactating hosts, having lower serum lactogen, alpha LA activity could be maintained in the absence of growth. In hypophysectomized hosts, even with continuous estrogen supplementation, alpha LA activity disappears and growth ceases, although ovine prolactin can replace the pituitary in restoring these traits. MCCLX  contains prolactin-binding capacity; thus, direct action of the lactogenic hormones on the tumor is possible. Our results indicate that MCCLX is a stably hormone-responsive tumor in terms of growth and differentiation.


Development of an agar-methyl cellulose clonogenic assay for cells in transitional cell carcinoma of the human bladder.

We report the development of a clonogenic assay for progenitor cells in transitional cell carcinoma of the bladder. Colony growth has been demonstrated from cells obtained both from surgical biopsies and from bladder barbotages. Electron microscopic and karyotypic evidence supports the contention that these progenitors represent a part of the population maintaining the tumor in vivo. Colony growth occurred in 9 of 11 surgical biopsy samples and in 6 of 6 bladder barbotage samples. Plating efficiency ranged up to 0.7%, and colony size was in some instances greater than 1000 cells. The assay appears potentially useful for analysis of the biology of human transitional cell carcinoma.


Assay for microsomal alpha-hydroxylation of N'-nitrosonornicotine and determination of the deuterium isotope effect for alpha-hydroxylation.

A high-pressure liquid chromatographic assay was developed for microsomal alpha-hydroxylation (2'-hydroxylation and 5'-hydroxylation) of N'-nitrosonornicotine. N'-Nitrosonornicotine was incubated with rat liver microsomes and a reduced nicotinamide adenine dinucleotide phosphate-generating system at 37 degrees. After addition of 2,4-dinitrophenylhydrazine reagent, the mixtures were analyzed by reverse-phase high-pressure liquid chromatography. The 2,4-dinitrophenylhydrazones of 4-hydroxyl-1-(3-pyridyl)-1-butanone and 4-hydroxy-1-(3-pyridyl)-butanal, which are the products of 2'-hydroxylation and 5'-hydroxylation, were quantified by ultraviolet light detection at 254 nm. Km's for 2'-hydroxylation and 5'-hydroxylation of N'-nitrosonornicotine by liver microsomes from Aroclor-treated male F-344 rats were 1.81 and 1.96 mM, while Vmax's were 0.53 and 1.05 nmol/min/mg protein, respectively. Aroclor pretreatment of rats resulted in a 20-fold induction of 2'-hydroxylation, but only a 1.9-fold induction of 5'-hydroxylation. The deuterium isotope effect for alpha-hydroxylation of N'-nitrosonornicotine was determined by comparing the rates of 2'-hydroxylation and 5'-hydroxylation of N'-nitrosonornicotine, N'-[2',5',5'-D]nitrosonornicotine, N'-[2'-D]nitrosonornicotine, and N'-[5',5'-D]nitrosonornicotine. The deuterium isotope effect (Vmax H/Vmax D) was 2.4 to 2.7 for 5'-hydroxylation and 1.2 for 2'-hydroxylation.


Mathematical models and the statistical analyses of cell transformation experiments.

Statistical methods are developed and applied to the fitting of the one-hit curves to chemically induced transformations in Syrian hamster colonies. Data from several experiments in two laboratories are shown to fit the model well at all but the highest doses of benzo(a)pyrene. The ratio of the estimated parameters of the one-hit curves ("transformicities") is proposed as a measure of the enhancement by X-irradiation. Statistical tests for possible nonrandom variation among dishes are developed for the situation where a sample of dishes is used to estimate the total number of surviving cells. The methods of fitting and estimation are also extended to this case. Further analyses exclude selection as an explanation for the observed increase in transformation frequency with increased carcinogen concentration. The reproducibility of the fit of the one-hit model and the radiation enhancement measures is ascribed to the uniqueness of this in vitro system in which the direct target cell-insult interaction is not modulated by host intervention.


Substrate specificity of human ribonucleotide reductase from Molt-4F cells.

Nucleoside triphosphates were examined as the activator for various nucleoside diphosphate reductions catalyzed by a highly purified ribonucleotide reductase obtained from Molt-4F cultured human cells. It was found that cytidine 5'-diphosphate and uridine diphosphate reductions are activated by adenosine 5'-triphosphate with apparent Ka's of 0.63 +/- 0.03 (S.E.) and 1.25 +/- 0.10 mM, respectively. Guanosine 5' diphosphate reduction is activated by deoxythymidine 5'-triphosphate with an apparent Ka of 1.25 +/- 0.11 microM, and adenosine 5'-diphosphate reduction is activated by guanosine 5'-triphosphate or deoxyguanosine 5'-triphosphate with an apparent Ka of 1.1 +/- 0.09 or 1.1 +/- 0.08 mM, respectively. In the presence of saturating amounts of their best activating nucleoside triphosphates, the Km's of various nucleotide diphosphates for this purified enzyme were studied. Double reciprocal plots of velocity against substrate concentration were found to be linear for all four substrates in the concentration range tested and yielded apparent Km's of 7 +/- 0.3 microM for cytidine 5'-diphosphate, 80 +/- 6.5 microM for adenosine 5'-diphosphate, 33 +/- 3.1 microM for guanosine 5'-diphosphate, 50 +/- 2.0 microM for uridine 5'-diphosphate. The reduction of one ribonucleoside diphosphate could be inhibited by other ribonucleoside diphosphates in a noncompetitive manner.


Diet and urinary steroids in black and white North American men and black South African men.

Urinary steroid hormone content was determined in Black and White North American men and in rural Black South African men between 40 and 55 years of age and in Black South African men over 60 years of age when maintained on their customary diets or when transferred to a vegetarian or Western diet, respectively. When eating their customary diets, Black South African men had lower levels of urinary estrogens and androgens than did Black and White North American men. The total androgen content decreased significantly in Black North American men on the vegetarian diet and increased in Black South African men fed a Western diet. Urinary excretion of estrogens was higher in older than in younger rural Black South African men. Data indicated that a vegetarian diet modified androgen and estrogen metabolism in North American men and that a Western diet was associated with higher levels of urinary steroid hormones in young Black South African men. Diet-related changes in steroid metabolism in rural Black South African men were age dependent. The relationship of the increased urinary excretion of steroid hormones in Black South African men, a low-risk group fed a Western diet, and the decreased excretion in Black and White North American men, high-risk groups fed a vegetarian diet, to the development of prostatic cancer remains to be clarified.


Effect of a bacteriocin produced by Mycobacterium smegmatis on growth of cultured tumor and normal cells.

Growth-inhibitory effects of a partially purified bacteriocin derived from Mycobacterium smegmatis ATCC 14468 on various animal cells transformed by tumor viruses, human malignant cells, and normal cells in the same species were studied. A growth-inhibitory effect of the bacteriocin on these cultured cells was determined by counting the residual cells. The bacteriocin inhibited virally transformed animal cells (mKS-A TU-7, 155-4 T2, and XC cells) and human malignant cells (AS-II and HGC-27 cells). The inhibitory effect increased with an increase in the bacteriocin activity. The bacteriocin sensitivities of transformed animal cells were relatively higher than were those of human malignant cells, while normal cells in the same species were practically insensitive to the bacteriocin. Differences in the degree of bacteriocin sensitivity were observed among tumor cell lines. Simian virus (SV) 40-transformed hamster cells (TSV-5 cells), which grow rapidly, were less sensitive to the bacteriocin. The cell membrane of SV40-transformed BALB/c mouse cells (mKS-A TU-7 cells) adsorbed the bacteriocin much more than did the cell membrane of nontransformed BALB/3T3 cells. The results seem to indicate that the inhibitory effect of bacteriocin 14468 on cultured mammalian cells probably depends on the binding sites for the bacteriocin which appear or increase by malignant transformation on cytoplasmic membrane.


A correlation between estrogen sulfotransferase levels and estrogen receptor status in human primary breast carcinoma.

Estrogen sulfotransferase (EC 2.8.2.4) activity and estrogen receptor levels were measured in 32 human primary breast cancer cytosol preparations. Two types of tumors were identified: type 1, in which estrogen sulfotransferase levels were low (less than 40 pmol 17 beta-estradiol 3-sulfate formed per mg protein per 2 hr) and were independent of [35S]adenosine 3'-phosphate 5'-phosphosulfate production from [35S]sulfate and adenosine triphosphate, and type 2, in which estrogen sulfotransferase levels ranged from 50 to 200 pmol 17 beta-estradiol 3-sulfate per mg protein per 2 hr and were correlated with [35S]adenosine 3'-phosphate 5'-phosphosulfate formation (r = 0.70; p less than 0.005). In type 1 tumors, 11 of 16 were estrogen receptor negative; in type 2 tumors, 2 of 16 were receptor negative. Estrogen sulfotransferase levels in receptor-negative tumors were significantly lower than the levels in receptor-positive tumors (p = 0.025).


Alteration in binding of dexamethasone to glucocorticoid receptors in regenerating rat liver after partial hepatectomy.

Changes in binding of dexamethasone (9 alpha-fluoro-11 beta, 17 alpha, 21-trihydroxy-16 alpha-methylpregna-1,4-diene-3,20-dione) to its receptors in regenerating rat liver after 70% hepatectomy were examined. Specific receptors for dexamethasone in the liver remnants of adrenalectomized rats decreased significantly during the period of DNA synthesis after 70% hepatectomy; then, they increased to above the control values between Days 4 and 7 after partial hepatectomy and subsequently returned to the control values. During the period of DNA synthesis, decreased binding was observed in partially hepatectomized rats with or without adrenals, but later enhanced binding was not prominent in rats with adrenals.


Establishment of a cell culture line from a transplantable rat stomach cancer induced by N-methyl-N'-nitro-N-nitrosoguanidine.

A cell line designated as BV9 was established in culture from a transplantable rat stomach cancer, which was originally induced in the glandular stomach of a Wistar rat by p.o. administration of N-methyl-N'-nitro-N-nitrosoguanidine. The monolayer cells, which have been subcultured for more than 35 passages, revealed pleomorphic features. Chromosomal analysis showed hypertetraploidy (mode, 95), and marker chromosomes were present. Cultured cells were injected s.c. into cyclophosphamide-conditioned syngeneic rats and produced tubular adenocarcinomas resembling the original tumor.


Lack of proportionality between rate of cell division and induction of tumors in carcinogen-exposed regenerating livers.

Although a single pulse of dimethylnitrosamine administered during the regenerative response of liver subsequent to 70% hepatectomy resulted in more primary hepatocellular carcinomas in treated livers than in controls, the response was not proportionate to the level of cell division. Further, the use of 55-g male Sprague-Dawley rats that displayed an extremely active regenerative response did not significantly shorten the lag period before the appearance of tumors. Of additional interest was the finding that the post-S period might be even more susceptible to dimethylnitrosamine than the S phase was. These results support the suggestion that many aspects of the interaction between carcinogens and dividing cells and the requirement for subsequent events for the development remain unclear.


Antitumor activity of cis-dichlorodiammineplatinum(II).

cis-Dichlorodiammineplatinum(II) has shown good broad-spectrum activity in the current Division of Cancer Treatment (National Cancer Institute) tumor panel. It meets Decision Network criteria for activity in at least five of ten tumor systems (the ip B16 melanoma, the sc CD8F1 mammary carcinoma, the ip colon tumor 26, the ip L1210 leukemia, and the ip P388 leukemia). Activity against the sc colon tumor 38, the iv Lewis lung tumor, the ic ependymoblastoma, and the human breast tumor xenograft (MX-1) was marginal. No activity was detected against the human lung tumor xenograft (LX-1), but good, reproducible activity was observed against the murine M5076 ovarian carcinoma. No schedule-dependency was observed after ip administration against the ip L1210 leukemia. The recently developed subrenal capsule assay offers promise as a rapid way (less than or equal to 11 days) of ranking the sensitivity of a variety of human tumors to cis-dichlorodiammineplatinum(II) and other chemotherapeutic agents.


Disposition and distribution of platinum following parenteral administration of cis-dichlorodiammineplatinum(II) to animals.

After iv administration of cis-dichlorodiammineplatinum(II) (cis-platinum) to animals, plasma levels of platinum decline in a biphasic manner, with a distribution phase (alpha) half-life of minutes and an elimination phase (beta) half-life of days. Urinary excretion of platinum is extensive on the first day after drug administration with a final urinary recovery of 70%--90% of the administered dose. Platinum is initially distributed to nearly all tissues with the highest levels appearing in kidney, liver, ovary, uterus, skin, and bone. There is no preferential uptake of platinum into tumor, although the presence of a tumor may alter the rate of platinum excretion and the extent of whole-body retention. No effect is seen on hepatic microsomal drug metabolism after ip administration of cis-platinum to rats. Platinum is excreted more rapidly from hydrated animals than from controls although total urinary recovery of platinum is nearly equal in both groups. Most analogs of cis-platinum appear to follow the same elimination and distribution patterns as cis-platinum itself.


Rationale for development of platinum analogs.

There is a great need for clinical trial of new second-generation platinum coordination compounds that might demonstrate greater clinical activity in a broader spectrum of tumors, decreased renal toxicity and emesis, improved solubility, synergism in combination therapy, and lack of cross-resistance to cis-dichlorodiammineplatinum(II) (cis-platinum). Lack of cross-resistance to cis-platinum is shown by certain 1,2-diamino-saturated cyclic platinum derivatives which also have a high degree of activity against transplanted mouse leukemias. cis-Platinum and these cyclic compounds combine synergistically with derivatives of cytosine arabinoside, VP-16-213, and Adriamycin. These 1,2-diamino cyclic compounds appear to have less renal toxicity than cis-platinum. The toxic and therapeutic effects of both cis-platinum and the diamino cyclic compounds can be blocked by massive doses of thiourea. Varying doses and time intervals of thiourea rescue are being studied in the hope of improving the therapeutic index of the platinum derivatives.


Antileukemia (L1210) activity and toxicity of cis-dichlorodiammineplatinum(II) analogs.

cis-Dichlorodiammineplatinum(II) (cis-platinum) and 40 of its analogs were evaluated for antitumor activity in BDF1 mice implanted ip with 10(6) L1210 leukemia cells. Several of these analogs were also evaluated for their ability to cause elevation in BUN and to produce leukopenia in normal BDF1 mice. In 11 experiments, cis-platinum given on Day 1 or Days 1--9 after implant produced T/C (treated/control) values between 164% and 214% and 157% and 285% respectively. On the basis of single experiments, 13 analogs were judged to be comparable to cis-platinum in that they produced T/C values greater than or equal to 167% after a single dose or greater than or equal to 200% after daily doses for 9 days. These active compounds included various substituted amine derivatives of dichloroplatinum(II), malonatoplatinum(II), aquasulfatoplatinum(II), and chloroacetatoplatinum(II) and derivatives of dihydroxydichloroplatinum(IV). Twelve of these active analogs and cis-platinum were evaluated for toxicity at doses ranging from near the optimal therapeutic dose to greater than or equal to the LD50. Only five of the 12 analogs and cis-platinum caused an increase in BUN to greater than 30 mg/100 ml, while eight of the analogs produced leukopenia comparable in incidence and severity to that produced by cis-platinum.


Pharmacokinetics of non-protein-bound platinum species following administration of cis-dichlorodiammineplatinum(II).

The pharmacokinetics of non-protein-bound platinum species derived from cis-dichlorodiammineplatinum(II) (cis-platinum) was studied under a variety of dosing conditions. Following rapid infusions (15-minute) of cis-platinum at 100 mg/m2, the unbound drug declined in a biphasic mode with a mean terminal half-life of 48 minutes. The mean beta-phase half-life after a 6-hour infusion of the same dose of cis-platinum was 26 minutes. Urinary excretion of filterable platinum was substantially greater after a 6-hour infusion than after a 15-minute injection. Concomitant administration of mannitol appeared to result in higher peak plasma concentrations and decreased urinary excretion of unbound platinum species but did not alter the terminal half-life. Renal impairment was associated with extremely high plasma levels of filterable platinum but did not affect other pharmacokinetic parameters. Preliminary data on the distribution of cis-platinum to ascitic fluid are also presented.


cis-Dichlorodiammineplatinum(II) in the treatment of gynecologic malignancies: phase II trials by the Gynecologic Oncology Group.

A dose of 50 mg/m2 of cis-dichlorodiammineplatinum(II) was administered iv every 3 weeks to 25 patients with squamous cell carcinoma of the cervix and to 34 patients with adenocarcinoma of the ovary. All patients had advanced or recurrent disease. Eleven of the 25 patients (44%) with cervical carcinoma had an objective response while ten of 34 patients (29%) with ovarian carcinoma demonstrated an objective response. Toxic effects were frequent but tolerable, with no drug-related deaths and only minimal drug-related nephrotoxicity, except in one patient. Myelosuppression was more frequent in the more heavily previously treated ovarian carcinoma patients. cis-Dichlorodiammineplatinum(II) thus appears to be a highly active single agent in the treatment of squamous cell carcinoma of the cervix and adenocarcinoma of the ovary.


cis-Dichlorodiammineplatinum(II)-based chemotherapy as initial treatment of advanced head and neck cancer.

During the past 2--3 years, a total of 73 patients with advanced head and neck cancer (mostly stage IV) were treated with cis-dichlorodiammineplatinum(II) (cis-platinum) as initial therapy, either alone or in combination with other agents. After chemotherapy, the patients received surgery and/or radiation therapy in standard fashion. Major degrees of tumor regression were seen in 40%--71% of the patients. cis-Platinum and bleomycin given by infusion gave the highest response rate (71%). The addition of high-dose methotrexate to cis-platinum plus bleomycin produced an unacceptable level of toxicity. Similarly, a four-drug regimen in which low-dose methotrexate and vinblastine were added to cis-platinum and bleomycin resulted in increased toxicity without additional therapeutic effect. Initial chemotherapy does not compromise subsequent surgery or increase either the immediate complications of surgery or the acute toxicity of radiation therapy. It seems unlikely that initial chemotherapy has altered the grim prognosis of an advanced, inoperable presentation. The effect of initial chemotherapy on advanced, operable disease will have to be assessed by suitable controlled trials.


Advanced bladder cancer: therapy with cis-dichlorodiammineplatinum(II), adriamycin, and 5-fluorouracil.

Nineteen patients with unresectable or metastatic urothelial cancer were treated with the combination of cis-dichlorodiammineplatinum(II), Adriamycin, and 5-fluorouracil. One patient died within 2 weeks of therapy and one patient had no measurable disease, leaving 18 patients evaluable for survival and toxicity and 17 patients evaluable for response. Eleven patients (65%) achieved a partial remission with attendant clinical improvement, with a median duration of 25 weeks. All nonresponders have died, with a median survival of 17 weeks. The median survival of responding patients was 40 weeks, with two patients alive at 33+ and 63+ weeks. Of particular interest are three patients who had resections of residual pelvic disease after significant regression with chemotherapy. One of these patients remains disease-free at 63+ weeks. Toxicity was significant but generally manageable.


Phase II evaluation of cis-dichlorodiammineplatinum(II) in advanced malignancies of the genitourinary and gynecologic organs: a Southwest Oncology Group Study.

One hundred and forty-seven fully and partially evaluable patients with advanced measurable malignancies of the genitourinary and gynecologic organs were given cis-dichlorodiammineplatinum(II) at a dose of 75 mg/m2 iv every 3 weeks. Thirty-six patients with testicular neoplasms were studied; five complete responses (13.9%) and seven partial responses (PR) (19.4%) were noted. Thirty-seven patients with ovarian adenocarcinoma were evaluated; five PRs (13.5%) were seen. One complete response (11.1%) and two PRs (22.2%) were obtained among nine patients with urinary bladder cancer. Four PRs (19.0%) were seen among a group of 21 patients with advanced prostate cancer. One PR (4.8%) was noted among 21 patients with renal cell cancer and no responses were seen in eight patients with cervical cancer. There was a highly statistically significant (P less than 0.001) survival advantage for the responding testicular tumor patients. Toxicity was similar to that previously reported, with gastrointestinal side effects and nephrotoxicity most commonly seen. Prospective and sequential analysis of renal function provided strong evidence for cumulative nephrotoxicity in these patients given bolus injections of cis-dichlorodiammineplatinum(II) without prehydration or treatment with fuosemide or mannitol.


cis-Dichlorodiammineplatinum(II) in non-small cell carcinoma of the lung.

The role of cis-dichlorodiammineplatinum(II) (cis-platinum) as a single agent in non-small cell lung cancer has not been clearly defined, and extensive phase II studies have not been conducted. However, objective responses were noted in several early clinical trials. A review of these reports, including combinations with conventional agents, shows 15 partial responses among 100 patients. Results are presented on a new protocol using high-dose cis-platinum with mannitol-induced diuresis combined with cyclophosphamide and Adriamycin. An overall partial response rate of 28% was observed, including a 38% response rate in good performance status patients, with a median survival of 16 months in responding patients. Data from recent combination protocols do not define the activity of cis-platinum as a single agent, but suggest that it may be useful in non-small cell carcinoma of the lung.


Phase II evaluation of cis-dichlorodiammineplatinum(II) in lymphomas: a Southwest Oncology Group Study.

The Southwest Oncology Group has evaluated the activity of cis-dichlorodiammineplatinum(II) at a dose of 75 mg/m2 given as an iv bolus injection every 3 weeks to 25 fully and partially evaluable patients with advanced Hodgkin's disease and non-Hodgkin's lymphoma. One complete response, two partial responses, and one improvement less than a partial response were noted. Myelosuppression, in the form of leukopenia and thrombocytopenia, was identified and seemed to be more prevalent and more severe than in previous studies. We have attributed this to the extensive prior treatments which these patients had received and to the presence of tumor-bearing marrow which was observed in some of them. The anticipated toxic effects which were noted included nausea and vomiting, anorexia, diarrhea, renal injury, and hyperuricemia. The precise role of cis-dichlorodiammineplatinum(II) in the management of human lymphomas awaits elucidation.


cis-Dichlorodiamminepltinum(II) in adult patients: Southwest Oncology Group Studies.

In addition to cis-dichlorodiammineplatinum(II) studies in lymphomas and testicular cancer and a broader phase II study of all genitourinary malignancies, the Southwest Oncology Group is currently conducting eight other studies which examine the effectiveness of this drug in adult patients with a variety of tumor types. This report will give preliminary analyses of the five studies for which there are early data, plus briefly describe the design of the three remaining protocols for which there are no data as yet.


Actin is unevenly distributed in the pituitary gland.

In view of the suggestion that actin-like proteins might be involved in the final steps leading to hormone secretion, the actin content of pituitary glands of adult rats was determined by sodium dodecylsulfate-polyacrylamide gel electrophoresis (for total actin), by the DNAse method (which measures predominantly monomeric actin) and by immunocytochemistry. The amount of actin present in the neural lobe, expressed per mg total protein, was found to be comparable to that of other neural tissues. In contrast, in the anterior lobe, the ratio was significantly lower. The intensity of immunofluorescent staining with anti-actin antibodies was higher in the neural lobe than in either anterior or intermediate lobes. The intensity and distribution of tubulin immunofluorescent staining with anti-tubulin antibodies resembled that of anti-actin antibodies. Thus, three independent methods point to an uneven distribution of actin in the subdivisions of the pituitary gland, although all these subdivisions are believed to secrete their hormones by exocytosis. These data suggest that the bulk of actin present in pituitary cells is unlikely to be involved only in exocytosis, but may be implicated also in the intracellular translocation of secretory products.


Cyto- and synaptogenesis in the arcuate nucleus of the rat hypothalamus during fetal and early postnatal life.

Morphogenesis of the arcuate nucleus of the rat from the 15th fetal day to the 6th postnatal day was investigated light and electron microscopically. The arcuate neurons exhibit a gradual development after the 15th fetal day. All cytoplasmic constituents are present in these nerve cells already during the last days of gestation. Nevertheless, they are not fully differentiated at birth. The first synapse-like structures (presynapses) were observed in 17 day-old, the first synapses in 18 day-old fetuses. During the early postnatal period the number of presynapses decreases, but at the same time there is a gradual increase in the number of the relatively mature synapses. This process starts already during the last days of prenatal life. Although all structural elements of the arcuate nucleus of the adult rat appear to be present at birth, the extent of the neuropil area and the number of ther presynapses indicate that the arcuate nucleus is still in a fairly undeveloped stage during the first postnatal days.


Light- and electron-microscopic structure of cells protruding into the mesencephalic ventricle of Scyllium stellare (Elasmobranchii, Selachii).

In the elasmobranch fish, Scyllium stellare, a complex group of cells protrudes into the cavity of the mesencephalic ventricle of the optic tectum. It consists of six to seven large spherical perikarya which resemble neurons of the mesencephalic nucleus of the Vth cranial nerve. The bundled processes of these cells form a stalk connecting the protrusion with the brain tissue. The protrusion is located in the region where the mesencephalic ventricle joins the cerebral aqueduct. This complex was not found in all specimens examined in the present study. The functional role of this peculiar group of cells, which contain dense core granules and are bathed in the cerebrospinal fluid, is open to discussion.


Observations on the origins of plasma membrane in rat deciduoma.

An NADH-ferricyanide reductase activity resistant to inactivation by cytochemical procedures was examined during decidualization of rat endometrium. Resistant activity was restricted to plasma membranes, distal elements of the Golgi apparatus, and discoid cisternae and cytoplasmic vesicles of decidual cells of endometrium of the pseudopregnant rat on days 3, 4, 5, 7, and 9, after mating. The procedure reduced or eliminated any evidence of NADH-ferricyanide reductase activity from other cellular components such as endoplasmic reticulum, nuclei, and mitochondria. The observations of the glutaraldehyde-resistant reductase in both plasma membranes and discoid cisternae may indicate a role for the latter in the biosynthesis of plasm membranes during decidualization when massive cell proliferation and membrane biosynthesis occur. The origin of the discoid cisternae is tentatively ascribed to the mature faces of the Golgi apparatus.


Fine structure of the ependymal cells in the area postrema of the domestic fowl.

The ultrastructure of the ependymal cells in the area postrema of the domestic fowl was studied by scanning and transmission electron microscopy. The ependymal surface of the area postrema is covered with many furrows and ridges. These ridges consist of ependymal cells aggregated in a fan-like shape. The ependymal cell lacks clustered cilia, microvilli are few, and a long basal process extends through the parenchymal layer of the area postrema. Within the cytoplasm as well as in the basal process, a spherical body with a diameter ranging from 1.5 to 2 micron is occasionally observed.


Development of the olfactory organ in the rainbow fish Nematocentris maccullochi (Atheriniformes, Melanotaeniidae).

The development of the olfactory organ in the rainbow fish, Nematocentris maccullochi, was studied using scanning and transmission electron microscopy; it was compared with the developmental process in other teleosts, especially in the closely related atherinids and cyprinodonts. The formation of the nares parallels that in atherinids, salmonids, cyprinids and heterosomats, but differs from that found in cyprinodonts. Another ontogenetic feature in which the olfactory organs of the rainbow fish and also of atherinids differ from those of cyprinodonts, is the occurrence of transitory kinociliary cells which disappear during the postlarval period. The divergent evolutionary pathways are discussed with reference to experimental investigations. During development, ciliated and microvillous receptor cell types occur. At the primary larval stage ciliated receptor neurons are exclusively present. At a later stage the microvillous type develops and becomes equal in frequency. Thus, the microvillous receptor represents a separate type of olfactory neuron and is not a progenitor of the ciliated receptor cell.


Biosynthesis of the basal lamina as a result of interaction between thyroid and mesenchymal cells in culture.

Porcine thyroid cells were cultured alone or in mixed cultures with mesenchymal cells. The formation of a basal lamina in vitro was investigated ultrastructurally. Follicular reassociation of thyroid cells occurred in both types of culture; however, it was followed by formation of the basal lamina only when mesenchymal cells were present. The present findings suggest an epithelial origin of the basal lamina resulting from an interaction with mesenchymal cells.


Cine-densitometric measurement of coronary arterial stenoses.

Computer-aided operator-interactive densitometry has been developed and applied to determine the percent stenosis for obstructive lesions in the coronary arterial tree. Phantom experiments performed to assess system linearity, accuracy of densitometric measurements, nonuniformity in image amplified response, and the interference of structure noise in measuring precision have been described. Three observers assessed the reproducibility of the method by repeating analyses on 20 stenoses in man recorded on 35-mm cine angiograms. Calculation of the intra- and interobserver errors found that their values significantly decrease with increasing percent stenosis. The reproducibility of the technique was also tested on stenoses that were exposed at multiple angles. The results agreed with those found for the intra- and interoperator errors. Limitations of the densitometric approach and desirability of further automation of the measurements are also discussed.


Evaluation of segmental left ventricular wall motion by equilibrium gated radionuclide ventriculography.

The ability of equilibrium gated radionuclide ventriculography to detect segmental left ventricular (LV) wall motion abnormalities was determined in 26 patients undergoing cardiac catheterization. Multiple gated studies obtained in 30 degrees right anterior oblique and 45 degrees left anterior oblique projections, played back in a movie format, were compared to the corresponding LV ventriculograms. The LV wall in the two projections was divided into eight segments. Each segment was graded as normal, hypokinetic, akinetic, dyskinetic, or indeterminate. Thirteen percent of the segments in the gated images were indeterminate; 24 out of 27 of these were proximal or distal inferior wall segments. There was exact agreement in 86% of the remaining segments. The sensitivity of the radionuclide technique for detecting normal versus any abnormal wall motion was 71%, with a specificity of 99%. Equilibrium gated ventriculography is an excellent noninvasive technique for evaluating segmental LV wall motion. It is least reliable in assessing the proximal inferior wall and interventricular septum.


Pulmonary atresia and suprasternal echocardiography.

A 54-year-old housewife with pulmonary atresia and ventricular septal defect developed increasing cyanosis, dyspnea, weakness, and myocardial ischemia. Initial cardiac catheterizations with angiography, including subtraction techniques, did not definitely indicate main pulmonary artery or right or left pulmonary arteries that would make her amenable to currently available surgical correction. Single-crystal suprasternal notch echocardiography indicated the presence of a small but definite right pulmonary artery. She subsequently underwent first-stage surgical correction after angiography and selective catheterization of aortopulmonary shunt vessels, which also revealed the presence of small proximal pulmonary arteries.


Foreign body retrieval from the heart by two catheter technique.

Foreign bodies that lodge accidentally in the heart or great vessels can be retrieved successfully without thoracotomy. This report describes retrieval of a 15-centimeter-long central venous catheter lodged in the right side of the heart. Two catheters were introduced to effect retrieval, one from the arm and the other from the leg. To our knowledge this approach has not been described previously.