The topography of the superificial roots and ganglia of the anterior lateral line nerve of the smooth dogfish, Mustelus canis. The topographic relationships of the superficial roots and ganglia of the anterior lateral line nerve (NLLa) to each other and to cranial nerves V, VII and VIII were studied, in the smooth dogfish Mustelus canis, by dissection of Sudan Black B stained specimens. NLLa consists of four branches: namely, superficial ophthalmic, buccal, otic and external mandibular. Each branch carriers lateral line neurons exclusively and forms a dorsal root and a ventral root which enter the anterior lateral line lobe and posterior lateral line lobe of the medulla respectively. It is estimated that slightly more than 50% of the fibers of the superficial ophthalmic, approximately 60% of the fibers of the buccal, at least 35% of the fibers of the otic and about 20% of the fibers of the external mandibular constitute the dorsal root. The ventral root is comprised of less than 50% of the fibers of the superficial ophthalmic, 40% of the fibers of the buccal, at least 50% of the fibers of the otic, and 80% of the fibers of the external mandibular. These results are correlated with the peripheral distribution and central termination of NLLa and it is concluded that the dorsal root carries axons from ampullary receptors and the ventral root carries fibers that innervate ordinary lateral line sense organs.


The efferent connections of the olfactory bulb and accessory olfactory bulb in the snakes, Thamnophis sirtalis and Thamnophis radix. The efferent connections of the olfactory bulb and accessory olfactory bulb of two species of garter snakes, Thamnophis sirtalis and T. radix were studied with experimental anterograde degeneration techniques. Axons of cells located in the olfactory bulb terminate ipsilaterally in all parts of the anterior olfactory nucleus, olfactory tubercle and lateral pallium. In addition, some axons enter the ipsilateral stria medullaris thalami, cross the midline in the habenular commissure, enter the contralateral stria medullaris thalami and terminate in the contralateral lateral pallium. The axons of cells in the accessory olfactory bulb course through the telencephalon completely separated from the fibers of olfactory bulb origin and terminate predominantly in the nucleus sphericus. These results confirm previous reports of the separation between the central projections of the olfactory and vomeronasal systems in a variety of vertebrates. The totality of the separation between these two systems coupled with the extensive development of the vomeronasal-accessory bulb system in these snakes suggests that they may be ideal subjects for further research on the functional significance of the vomeronasal system.


The structure and development of the pineal complex in the lanternfish Triphoturus mexicanus (family mycotphidae). The pineal complex of the lanterfish Triphoturus mexicans was studied by light and electron microscopy. Receptor, supportive and ganglion cells were identified in the pineal end-vesicle. The presence of receptor cells and the prominent pineal window strongly suggest a photoreceptor function. Numerous unmyelinated nerve fibers were also found in the end-vesicle. These converge to form part of the pineal stalk. Two kinds of cells (Types I and II) were distinguished in the dorsal sac, which is very well developed. Type I cells are non-ciliated ependymal cells and contain large parcels of glycogen. Type II cells contain large lipid-like inclusions and a distinct band of filaments around the entire periphery of the cell. Receptor cells appear very early in the development of T. mexicanus. They seem to originate from ependymal cells lining the roof of the third ventricle.


Karyotype, marker formation, and oncogenicity in mouse plasmacytomas. Two common chromosome markers in the 2 plasmacytomas previously examined by Giemsa banding were consistently present in the mouse plasmacytoma X-5563, a transplantable hypertetraploid tumor of spontaneous origin in C3H mice. The 2 markers were found in both induced and spontaneous tumors and in either BALB/c or C3H mice. The derived cell line had 17 fewer chromosomes than the X-5563 tumor and was oncogenic, and its modal karyotype was identical to that of the tumor transmitted by the inoculation of the cell line. The homogeneity of a slight karyotypic modification in a second tumor suggested a possible clonal origin of that tumor. The high frequency of centric fusions between homologues and the structure of certain markers suggests that homologue association may precede marker formation. We proposed a second mechanism of marker formation, selective regional elongation, to account for the larger number of markers with proximal or distal elongations without evidence of translocation and for the observed alterations in length and banding pattern of markers after growth in vitro. Comparison of MOPC-21, MOPC-315, and X-5563 tumors showed preferential involvement of certain chromosomes in marker formation, an inferred association of the 2 common markers with an early stage in the origin of the 3 plasmacytomas, and consistent loss of an X chromosome. Loss of oncogenicity in cell lines was associated with a number of karyotypic changes, but did not require the loss of the characteristic markers or additional copies of a specific normal chromosome.


Tumor-promoting activity of certain extracts of tobacco. Tumor-promoting activities of extracts of fluecured and cigarette tobacco were evaluated. Initially, fluecured tobacco was extracted consecutively with hexane, chloroform, acetone, ethyl alcohol, methyl alcohol, and water; and the extracts were tested for tumor-promoting activity on mouse skin. The hexane and chloroform extracts were fractionated on silicic acid, and the fractions were devoid of tumorigenic activity. The acetone and alcohol extracts showed marginal activity. Subsequently, chloroform-extracted cigarette tobacco was extracted with water, and the aqueous extract was partitioned by solvent precipitation methods. Bioassay results showed tumor-promoting activity for the aqueous extract, with tumors in 38% of the animals. The aqueous extract appeared about five times as active as smoke condensate derived from an equal weight of tobacco.


Specificity of the 5q-chromosome in a distinct type of refractory anemia. A further instance of the 5q-chromosome associated with a distinct type of aregenerative anemia strengthened the hypothesis of specificity and consistency of this karyotypic abnormality in a new hematologic entity. The Giemsa banding technique revealed that the deletion of the long arms of chromosome number 5 was interstitial: del (5) (pter leads to q12 : : q31 leads to qter).


Age-dependence of hepatic dimethylnitrosamine-demethylase activity in the rat. The mixed-function oxidase which activates the carcinogen dimethylnitrosamine (DMN) was determined in the rat liver as a function of animal age. DMN-demethylase activity increased considerably at first to reach a maximum on day 29, and then substantially decreased to day 59; thereafter, enzyme activity remained essentially stable up to at least day 110. Pretreatment with 3-methylcholanthrene, which caused a pronounced decrease in this enzyme activity, did not affect the general shape of the age-dependence curve. The results suggest that rats between weaning and sexual maturity are more susceptible to the carcinogenic effects of pulse doses of DMN than are neonates or adult animals.


Response of Cloudman S91 melanoma cells to melanocytestimulating hormone: enhancement by cytochalasin B. Cloudmann S 91 mouse melanoma cells treated for 5 days with melanocyte-stimulating hormone (MSH), cytochalasin B (CB), or both, exhibited changes in cell volume, population, nucleation, and pigment production. Cells treated with CB or CB in combination with MSH were eight to nine times larger, rounder, multinucleated, and heavily pigmented. CB alone increased melanin and DNA per nucleus threefold. CB in combination with MSH increased melanin per nucleus 30-fold. Data on DNA per nucleus suggest that CB-treated cells remained in the G phase longer than did control cells. MSH alone caused a reduction in cell population and a fourfold increase in melanin per nucleus.


Mantel-Haenszel analysis of Oxford data. I. Independent effects of several birth factors including fetal irradiation. Data from the Oxford Survey of Childhood Cancers were subjected to the Mantel-Haenszel procedures to recognize independent effects of associated factors in retrospective data. Our results showed that several birth factors, including fetal irradiation, social class, maternal age, and sibship position, exerted separate effects on childhood cancers in general, and reticuloendothelial system neoplasms in particular.


Epidemiology of diseases in adult males with leukemia. In the Tri-State Leukemia Survey, the history of diseases in 605 adult male leukemia cases 15 years and older and in 668 adult male population controls was examined. These diseases occurred at least 1 year before leukemia was diagnosed. The data were based on respondents' answers that the disease was diagnosed by a physician; the respondent was either the subject or his spouse. Of 30 diseases studied, 7 showed an excess among the patients with leukemia: infectious hepatitis, eczema, psoriasis, diabetes, arthritis and rheumatism, heart disease, and ankylosing spondylitis. Mumps had a lower reported occurrence among the cases, whereas pneumonia was less frequent in acute lymphatic cases than in population controls. Three diseases occurred significantly less in controls than in persons with specific histologic types of leukemia. Our data revealed a more frequent history of herpes zoster (shingles) in chronic lymphatic leukemia, more hives in acute chronic myeloid cases, and meningitis in acute myeloid leukemia. When we only considered the patients' responses, more of them admitted having had acne than did our controls. The remaining diseases--childhood viral diseases, infectious mononucleosis, smallpox, typhoid fever, dysentery, scarlet fever, tuberculosis, asthma, hay fever, and goiter did not occur more frequently in cases than in controls. The findings were consistent with evidence from previous laboratory and clinical studies. The increased occurrence of infectious hepatitis in our case series is consistent with the findings of other studies showing an increased frequency of Australia antigen in patients with hepatitis, leukemia, and Down's syndrome.


Prognostic significance of ploidy level in human tumors. I. Carcinoma of the uterus. The 5-year survival rates of 540 patients with carcinoma of the cervix and 186 with carcinoma of the corpus uteri were assessed in relation to the modal DNA values of the tumors. Patients with squamous cell cervical carcinomas had more favorable prognoses if the modes were near-triploid or hypotetraploid; however, these high-ploidy tumors included more stage III cases than did the tumors with near-diploid modes. Patients with near-diploid endometrial carcinomas has considerably more favorable prognosis than did patients with the minority of tumors at this site, who had high modes; this prognostic difference was only partly related to a higher proportion of poorly differentiated tumors in the high-ploidy group since, among the poorly differentiated tumors, individuals with near-diploid modes again had significantly better prognoses than those with high modes.


Type C RNA viruses and leukemogenesis: association of Gross strain of murine leukemia virus infection and leukemogenesis in rats. The correlation of murine leukemia virus (MuLV) infection and leukemogenesis was examined by XC plaque assay in W/F rats neonatally infected with the rat-adapted Gross virus (RAGV). Thymic lymphomas developed in 100% of infected rats, with a mean latent period of 64 days. The virus infectivity was first detected in the thymus as early as 15 days after the virus infection; the titer steadily increased thereafter until the development of thymic lymphoma. No other tissues of virus-infected rats showed virus infectivity until the development of localized thymic lymphoma, though sera of a few rats showed low infectivity titers. In rats with generalized leukemia, however, high titers were detected in the sera and leukemic tissues. In untreated controls, all tissues tested invariably showed negative titers for MuLV infectivity at any age up to 7 months after birth. The results indicated that those tissue sites were common to both RAGV infectivity and the leukemogenic process, with the primary involvement of the thymus, and that the appearance of RAGV infectivity in various tissues represented the expression of the oncogenic genome of RAGV.


Spontaneous tumors and common diseases in two colonies of Syrian hamsters. I. Incidence and sites. Syrian hamsters from two colonies [Eppley colony (EC); Hannover colony (HC)] were examined for spontaneous neoplastic and noneoplastic diseases. Significant differences were found in tumor incidence (30%, EC' 42%, HC), occurrence of malignant neoplasms (15%, EC; 32%, HC), appearance of multiple primary tumors in different organs (42%, EC; 30%, HC), average survival rates of tumor-bearing animals, organ distribution and histologic types, and average survival rates of all animals. The frequencies of nonneoplastic diseases also varied between the two colonies. The findings indicated a need for precise histologic evaluation to generate information for comparative purposes regarding the use of hamsters in experimental pathology and carcinogenesis studies.


Spontaneous tumors and common diseases in two colonies of Syrian hamsters. III. Urogenital system and endocrine glands. The frequency of spontaneously occuring neoplasms in the urogenital system and endocrine organs was examined in two Syrian hamster colonies [Eppley colony (EC); Hannover colony (HC)]. With the exception of a renal adenoma in an EC female, the urinary systems of animals in both colonies were free of neoplastic growths. A single epididymal adenoma of the male fenital system (EC) was found. However, incidences of female genital tract tumors were 10 (EC) and 3.5% (HC) and accounted for 34 and 11%, respectively, of all tumors in females. Among all genital tract tumors, 3 (EC) and 57% (HC) were ovarian, 14% (HC) were in the fallopian tubes, 81 (EC) AND 29% (HC) were uterine, and 16% (EC), vaginal. The overall incidence of endocrine organ tumors was similar in both colonies (EC, 22%; HC, 19%). The distribution of these tumors was as follows: pituitary gland; EC, 3%, HC, 0; thyroid gland: EC, 28%, HC, 31%; parathyroid gland: EC,11%, HC, 18%; adrenal cortex: EC, 42%, HC, 31%; adrenal medulla: EC, 7%, HC, 0; and endocrine pancreas: EC, 10%, HC, 31%. The morphology of these neoplasms is described and the incidence compared with those reported in other colonies.


Spontaneous tumors and common diseases in two colonies of Syrian hamsters. IV. Vascular and lymphatic systems and lesions of other sites. Spontaneous tumors were found in the vascular and lymphatic systems upon systemic histologic examination of the cardiovascular, hematopoietic, and lymphatic tissues of Syrian hamsters from the Eppley (EC) and Hannover colonies (HC). The incidence of endothelial tumors (hemangioendotheliomas) was similar in both colonies and sexes. The hemangioendotheliomas generally originated in the liver and spleen. Malignant lymphomas developed in high incidences in both colonies; however, their morphology varied. Those in EC hamsters were of histiocytic and, to a lesser extent, plasmacytic types; in the HC lymphocytic and epithelioid cell types were also found. Among tumors of miscellaneous sites, Harderian gland adenomas occurred in almost equal frequencies in EC and HC hamsters, but predominated in males. Tumors of the bone and soft tissue were present only in EC hamsters, whereas skin neoplasms developed solely in HC hamsters.


Specific cytotoxicity in vitro of lymphocytes sensitized in culture against tumor cells. The production of tumor-specific cell-mediated cytotoxicity following in vitro sensitization of C57BL spleen cells against a syngeneic 3LL Lewis lung carcinoma was studied. Lymphocytes were sensitized on monolayers of the tumor cells for 4-5 days. The cytotoxicity was assayed by measuring the reduction in 3H-leucine and 3H-thymidine incorporation by target cells after interaction with the sensitized lymphocytes. Spleen lymphocytes sensitized on monolayers of 3LL tumor cells caused a high extent of lysis; such cells tested on C57BL or C3H fibroblast targets evoked only a low level of cytotoxicity. C57BL spleen cells sensitized on C57BL fibroblasts caused a low level of cytotoxicity when tested on a 3LL target. Thus cytotoxicity appeared to be tumor specific. The reduced incorporation into protein and DNA of target tumor cells caused by the sensitized lymphocytes was a measure of cell injury, which was more sensitive than direct cell count or uptake of 51CR. Lymphocytes from syngeneic tumor-bearing mice, tested 13-25 days after tumor inoculation, did not manifest in vitro cytotoxicity. On the contrary, such lymphocytes sometimes appeared to have a promoting effect on the tumor cells.


Inhibitory effect of shark serum on the Lewis lung carcinoma. The growth and metastasis of transplanted Lewis lung carcinoma in C57BL/6J mice were inhibited by an ip injection of whole serum from Carcharhinus plumbeus, the sandbar shark. Tumors failed to develop in 69% of the animals inoculated with shark serum on days 0, 3, and 6 after tumor transplantation. Histologic examination of the tumor site at days 3 and 6 showed that tumor cells were pyknotic, and evidence of lysis of tumor cells and minor leukocytic infiltration existed. Tumor cells were not in tissue sections from day 15, and these animals still had no symptoms at day 216. The mean tumor volume of the remaining 31% of the treated animals was less than that of controls; they had a prolonged mean survival time, but ultimately they died from metastases, as did the controls. Urea- and hemoglobin-treated animals and those pretreated or intralesionally treated with shark serum were similar to the controls in both tumor kinetics and survival times.


Experimental cancer immunotherapy: comparison of tumor rejection if F344 rats given live Mycobacterium bovis (Strain BCG) and killed Corynebacterium parvum. F344 rats received grafts of syngeneic 13762 mammary adenocarcinoma cells previously admixed with either living BCG of killed Corynebacterium parvum administered sc or intradermally (id). Animals given id transplants of tumor cells admixed with either BCG or killed C. parvum exhibited tumor growth for an average of 10 days, then regression in size and rejection of the tumor nodules. Lesions were found in rats given sc transplants of tumor cells admixed with the killed microorganism for an average of 13 days with the same results. When live BCG was added to the sc transplants, accelerated rates of tumor growth and early death were noted, compared with the group receiving tumor cells alone sc. Suppressed rates of tumor growth and prolonged survival were observed in the groups receiving id inoculations of tumor cells followed by treatment with killed C. parvum administered weekly ip or id 1 cm away and around the growing tumor. On the other hand, weekly treatment of BCG injected either ip or id 1 cm away and around the growing tumor resulted in accelerated rates of tumor growth and early death. Animals exhibiting C. parvum of BCG-mediated tumor rejection displayed tumor-specific protection to sc challenge injections of the cell line initially used, but they died with growing tumors and metastases when challenged with tumor cells of an antigenically different line syngeneic to F344 RATS. Microscopic examination of histologic sections of tumors formed from id inoculations of tumor cells admixed with either BCG or killed C. parvum revealed a nonspecific infiltrate of macrophages and polymorphonuclear leukocytes in the tumor, whereas sections of tumors formed from sc grafts of cells admixed with killed C. parvum revealed a specific organized infiltrate of mostly macrophages around the tumor follicles.


Carcinogen-induced changes in tracheal epithelium cultured in serum-free, chemically defined medium. Rat tracheal tissue was cultured for periods up to 2 months in medium containing benzo[a]pyrene, and the epithelium was studied for the histologic effects of deletion of serum from the medium. Nodular hyperplasia occurred in 3 days in the absence of serum, but was not seen until several weeks of culture in media containing the same concentrations of carcinogen with 10% calf serum. In serum-free culture, hyperplasia was induced in 2 weeks with one-tenth of the smallest concentration of benzo[a]pyrene that yielded this change in serum-containing medium. The cells of the hyperplastic epithelium in both serum-containing and serum-free culture exhibited ultrastructural changes described in carcinogenesis in vivo. In the absence of serum, squamous hyperplasia was uniformly seen, a feature that distinguished serum-free culture from culture in the presence of calf serum. No frank intraepithelial or invasive malignant lesions were produced in either medium. It was concluded that exposure of organ cultures to carcinogen in the absence of serum is the more promising method for bioassay because the response to carcinogen was more rapid, more sensitive, and more reproducible than that seen during exposure in media that contained serum.


Efficient photolytic degradation of nitrosamines. The principles and practice of nitrosamine degradation by photochemical means were described. Two types of apparatus were constructed for this purpose. A nitrosamine waste solution containing 10-3-10-1 N in hydrochloric acid and about 10 mole-equivalents of an HNO2 scavenger (i.e., urea, guanidine, or hydrazoic acid) was efficiently and irreversibly degraded to the parent amines and N2 (and/or N2O) by irradiation. Efficiency of the degradation depended on the type of apparatus used. With the use of one apparatus, about 11 g N-nitrosodimethylamine was photolytically converted to dimethylamine, N2, and N2O in 24 hours. Protonated guanidine catalyzed the photohydrolysis processes.


Abnormal tryptophan metabolism in a family with a history of bladder cancer. Tryptophan metabolism was studied in a family in which a sister and brother had bladder cancer. Urinary tryptophan metabolites after administration of 2 g L-tryptophan were measured in 4 sisters and 1 brother, all free of disease when studied. One sister with and 2 without histories of bladder cancer had significantly elevated levels of kynurenine, acetylkynurenine, and 3-hydroxykynurenine. Administration of 100 mg pyridoxine hydrochloride returned the tryptophan metabolism to normal in these 3 individuals. One brother with a 1 sister without bladder cancer had normal metabolism. A repeat study 2 years later confirmed the abnormal metabolism in the 3 sisters. Two sisters with abnormal tryptophan metabolism were given 200 mg L-kynurenine sulfate orally to bypass the effects of tryptophan oxygenase activity. Both excreted elevated levels of kynurenine and 1 excreted elevated levels of 3-hydroxykynurenine.


Air-water and ether-water distribution of N-nitroso compounds: implications for laboratory safety, analytic methodology, and carcinogenicity for the rat esophagus, nose, and liver. The air-water distribution ratio K1, ether-water distribution ratio K2, and solubility in water were measured for 17 nitrosamines, 3 nitrosamides, and 1 nitrosocyanamide. For K1, air was analysed either by UV absorption of ethanol extracts or by gas chromatography. K1 at 37 degrees C varied from less than 2 X 10(-6) to 4.5 X 10(-3), with 11 compounds showing K1 greater than 10(-4). A literature analysis provided data on the carcinogenicity of these N-nitroso (NNO) compounds toward the esophagus, nose, and liver of the rat. This compilation of data on physical properties and carcinogenicity of NNO compounds was then analysed in terms of: a) safety of workers handling either solutions of volatile NNO compounds or animals treated with these compounds, b) analytic methdology, and c) possible correlations between K1-, K2-, and solubility-characteristics, and carcinogenicity. Overall correlations were not observed. However, within each of five chemical groups, K1 and K2 tended to be associated positively with esophageal and nasal carcinogenicity and negatively with hepatic carcinogenicity. Water solubility showed the opposite associations.


Human neoplastic and normal cells in tissue culture. I. Cell lines derived from malignant melanomas and normal melanocytes. Forty-six cell lines derived from 31 human melanomas obtained from 28 patients were cultured. Fourteen of 16 lines have produced malignant tumors when injected into nude (thymus-deficient) mice. Tumors in 5 of the nude mice metastasized to distant lymph nodes and/or to the lungs of the mouse host. Extreme variability from line to line was observed for doubling time (34 to 106 hr), plating efficiency (0-86%), and melanin production. All tested lines had type B glucose-6-phosphate dehydrogenase, thereby excluding HeLa cell contamination. HeLa cells have been grown for some time in our laboratory. Our results clearly demonstrated that HeLa cell contamination does not occur invariably in heteroploid lines growing in a laboratory simultaneously with Hela cells, provided that proper care is taken to avoid such occurrence. Multiple cell lines derived from the same tumor had identical phosphoglucomutase enzyme phenotype, which suggested a lack of significant cross-contamination between the lines. Four long-term cultures of normal human uveal embryo melanocytes have also been established and characterized. Although all produced melanin after reaching saturation density, they differed from the melanoma cells morphologically; they were flat, not refringent, and lacked piling up and plating ability. When melanoma cells were exposed to bromodeoxyuridine (BUDR) for long periods, a phenotypic change toward non-neoplastic characteristics was observed. Cells became flat and not refringent and, when injected into nude mice, tumors appeared after a long latent period. These changes were completely reversible in vitro and in vivo. The BUDR-treated cultures were undistinguishable from the untreated mother cultures after 2 to 3 passages. Lines derived from tumors in nude mice (obtained by injection of BUDR-treated cells) were again indistinguishable from the untreated mother line. Normal melanocytes were mostly euploid; all the melanoma cells were aneuploid. All 29 cell lines derived from 14 patients had an average chromosome number higher than 46. Detailed group-by-group chromosome analysis always showed an excess of C chromosomes, which suggested that hyperreduplication of one or more C chromosomes is a specific characteristic of human melanomas.


Control of cell growth. I. Estrogen-dependent growth in vivo of a rat pituitary tumor cell line. A subclone of rat pituitary tumor cells, designated GH3/C14, was isolated from the parent population of GH3 pituitary cells and was estrogen-dependent for growth in vivo. GH3/C14 cells inoculated into host animals formed tumors in intact females, estrogen-treated ovariectomized females, and estrogen-treated males, but not in untreated intact or castrated males, or untreated ovariectomized females. Exogenous treatment with estrogens supported tumor formation in male animals. Radioimmunoassay of the average serum estradiol concentrations that support tumor growth in intact females, estradiol-treated intact females, and estradiol-treated intact males gave values of 41 +/- 4, 1,130 +/- 150, and 730 +/- 140 pg/ml, respectively. Tumor formation by GH3/C14 cells inoculated into male animals was not supported by either exogenous progesterone or hydrocortisone acetate. Further, these two steroid hormones had no significant effect on the estrogen-promoted growth in males or females. Exogenous testosterone treatment promoted tumor formation in males and ovariectomized females, but dihydrotestosterone was completely ineffective. Radioimmunoassay of the serum from tumor-bearing animals and the medium from the cells in culture showed that the cells produced growth hormone in vivo and in vitro but did not produce measurable amounts of luteinizing hormone or follicle-stimulating hormone. The groth of GH3/C14 cells in culture was examined in medium without added estrogen, and with estradiol added to the level of either the normal intact female rat or the estradiol-treated animals. No direct growth stimulation by estrogens could be detected in culture; the data suggested an indirect growth control mechanism.


Control of cell growth. II. Requirement of thyroid hormones for the in vivo estrogen-dependent growth of rat pituitary tumor cells. Further examination of rat pituitary cell line GH3/C14 showed that at least the physiologic concentration of L-thyroxine was required for estrogen-dependent growth in vivo. Two L-thyroxine synthesis inhibitors, 6-n-propyl-2-thiouracil (propylthiouracil) and 1-methylimidazole-2-thiol (methimazole), were administered concurrently with estrogen to GH3/C14-inoculated hosts. Propylthiouracil administration to estrogen-treated males, intact females, and estrogen-treated ovariectomized females inhibited tumor formation by 93, greater than 95, and 68%, respectively, as compared to tumor formation in controls not treated with propylthiouracil. Methimazole treatment of estrogen-primed males and intact females inhibited tumor formation by 78 and 95%, respectively. Concentrations of total L-thyroxine and free L-thyroixine in sera from normal and inhibitor-treated hosts were depressed 70-80% by propylthiouracil and 60-70% by methimazole. Administration of either drug caused greater inhibition of tumor growth than of total body weight gain. In addition, the administration of a combination of L-thyroxine and L-triiodothyronine to male rats promoted tumor formation even in the absence of exogenous estrogen.


Control of cell growth. III. Direct mitogenic effect of thyroid hormones on an estrogen-dependent rat pituitary tumor cell line. A possible direct estrogen requirement for growth of GH3/C14 rat pituitary tumor cells was evaluated in culture medium supplemented with estrogen-depleted serum prepared by a 56 degree C charcoal extraction procedure, and with serum obtained from ovariectomized sheep and ovariectomized adrenalectomized sheep. Growth of the GH3/C14 cells in culture medium in which the final estrogen concentration was 2 pg/ml or less was equal to growth in medium with normal serum and equal to growth in the presence of estrogen-depleted serum to which estradiol was added back at concentrations of 10-1,000 pg/ml. Under no conditions could a direct estrogen requirement for growth be demonstrated. The function of thyroid hormones in cell growth was examined in culture medium supplemented with serum from thyroidectomized sheep. In such medium the growth of the GH3/C14 cells was stimulated 3.5-fold by addition of 1.0 X 10(-8) M L-thyroxine (T4) or 1.0 X 10(-9) m L-triiodothyronine (T3). Investigation of the possible synergistic effects of estrogens and T4 revealed that combinations of estrogen and T4 or T3 did not stimulate growth over that seen with T4 or T3 alone. These data indicated that estrogens are not direct growth requirements for these cells but instead operate in vivo through secondary or indirect mechanisms; in contrast, thyroid hormones are directly mitogenic in vitro.


Induction of tumors by a guinea pig herpesvirus-transformed hamster cell line. Syrian hamster embryo cells that had been transformed in vitro by guinea pig herpes-like virus (GPHLV) were found to be oncogenic when inoculated into hamster sc or ip. Of 71 animals inoculated, 30 showed tumors at the site of inoculation. Tumors appeared 4-23 weeks after inoculation of the transformed cells at passage 37 or higher. Inbred and randombred hamsters of all ages were susceptible. Upon microscopic examination the tumors were characterized as fibrosarcomas. The cultured hamster tumor cells were easily transplanted into hamsters, but produced no evidence of tumors when inoculated into guinea pigs. Infectious GPHLV was not isolated from the tumor cells, but GPHLV-specific surface antigens were detected in tumor cells by immunofluorescence of GPHLV antiserum produced in rabbits. Sera from tumor-bearing hamsters did not contain GPHLV-neutralizing antibodies, but sera from 4 of 23 hamsters bearing primary tumors and 12 of 41 bearing transplanted tumors produced nuclear fluorescence in cells infected with GPHLV, thus establishing the relationship between the guinea pig herpesvirus and the hamster tumors.


Antitumor activity of killed Corynebacterium parvum suspensions in a murine mammary adenocarcinoma CaD2) system. We studied the antitumor activity of killed Corynebacterium parvum on the CaD2 mammary adenocarcinoma in DBA/2 mice. Intratumor treatment had little or no effect on subcutaneous tumor growth. Admixture of tumor cells with C. parvum before inoculation completely suppressed tumor growth. No tumor transplantation immunity was detected in mice inoculated with admixtures of C. parvum and tumor cells, but tumors were enhanced under certain circumstances. Growth of a tumor inoculated sc or iv was consistently retarded after iv or ip C. Parvum therapy, but tumors rarely regressed. Tumor transplantation immunity was detected in mice treated iv with C. parvum before surgical excision of established tumors. Certain adverse effects of iv or ip administration of C. parvum were also discussed.


Leukemias and vaginal tumors induced in female Donryu rats by continuous administration of 1-butyl-3,3-dimethyl-1-nitrosourea in the drinking water. Three groups of Donryu rats, each consisting of 36 females, were continuously given solutions of 1-butyl-3,3-dimethyl-1-nitrosourea as drinking water (400 ppm for group A, 200 ppm for group B 100 ppm for group C). Of the 100 rats that survived at least 122 experimental days, 64 developed leukemia and 38 had vaginal tumors. Leukemias were preponderant in animals of groups A and B; vaginal tumors appeared in group C.


Sequential adsorption analysis of antibodies to neuroectodermal cells by an indirect quantitative method. An indirect radioimmunoassay was developed for the sequential adsorption analysis of rabbit antibodies raised against rat neuroectodermal cells. The quantitativee relationship between primary adsorbed rabbit antitumor antibodies and secondary adsorbed goat antibodies to rabbit IgG was explored by paired radioiodine analysis. In the final indirect method, the amount of unlabeled rabbit antibody removed by cultured monolayers of cells at each step in a sequence of cells could be determined from an equation relating the unlabeled amount to values of bound 125I-labeled goat antirabbit IgG. To obtain the total quantity of rabbit antibody in a particular antiserum reagent, a sequential adsorption analysis was done with successive steps of homologous cells. To obtain the amount of antibody that was cross-reactive for other cell lines, we included those lines in the first several steps of the sequence. The sequential adsorption profile was considered as a more important indicator of the quality of a particular antiserum reagent than was the total amount of antibody contained in it. Neuroectodermal cell lines used as illustrative examples included subclones of the C6 astrocytoma and of the RN-2 schwannoma.


A choroid plexus papilloma in an elasmobranch (Squalus acanthias). A choroid papilloma in the choroid plexus of the ala of the fourth ventricle in a mature male elasmobranch, Squalus acanthias, was described. This is apparently the first report of a neoplasm of the central nervous system in a member of the class Chondrichthyes.


Regulatory variants for the expression of H-2 antigens. I. Isolation and characterization. By subjecting "mutagenized" mouse myeloma cells repeatedly to selection with antiserum against H-2Kd, we isolated variants that expressed little or no H-2Kd alloantigens. The phenotype was unstable, with the culture accumulating revertants in the absence of selective pressure. The variants not only failed to express the selected antigen, but also the antigen coded by the closely liked H-2Dd gene. The cells reexpressed this antigen with the same kinetics as for the selected antigen. Several physiologic parameters (karyotype, cell morphology, size, and growth curves) were not altered by the loss of H-2 antigens.However,when injected iv, the H-2-deficient cells did not lodge in the spleen as readily as did the wild-type cells.


Cocarcinogenic and tumor-promoting agents in tobacco carcinogenesis. A series of 21 tobacco smoke components and related compounds werere applied to mouse skin (50 female ICR/Ha Swiss mice/group) three times weekly with a low dose (5 mug/application) of benzo[a]pyrene (B[a]P). The test compounds were of five classes: aliphatic hydrocarbons, aromatic hydrocarbons, phenols, and long-chain acids and alcohols. The following compounds enhanced remarkably the carcinogenicity of B[a]P: catechol, pyrogallol, decane, undecane, pyrene, benzo[e]pyrene, and fluoranthene. The following compounds inhibited B[a]P carcinogenicity completely: esculin, quercetin, squalene, and oleic acid. Phenol, eugenol, resorcinol, hydroquinone, hexadecane, and limonene partially inhibited B[a]P carcinogenicity. Six of the 21 compounds were also tested as tumor promoters im two-stage carcinogenesis. No direct correlation existed between tumor-promoting activity and cocarcinogenic activity. The cocarcinogens pyrogallol and catechol did not show tumor-promoting activity. Decane, tetradecane, anthralin, and phorbol myristate acetate showed both types of activity. Structure-activity relationships and possible modes of action were described.


Detection and estimation of azomethane in expired air of 1,2-dimethylhydrazine-treated rats. Azomethane (AM) gas was identified as a major metabolite of 1,2-dimethylhydrazine (1,2-DMH) in the expired air of F344 rats. The compound was characterized by high-pressure liquid chromatography, gas chromatography, and mass spectrometry, in comparison to a synthetic standard. At a dose of 21 mg 1,2-DMH/kg sc, approximately 14 and 11% of the dose were exhaled as AM and CO2, respectively, in 24 hours. At 200 mg 1,2-DMH/kg, 23 and 4% of the dose appeared as AM and CO2, respectively, in the respired air within the same period. Most AM was seen in the first 6 hours, but the CO2 evolution was more progressive, especially after the higher dose of 1,2-DMH.


Increased immunogenicity of TA3-Ha cells treated with the antitumor antibiotic macromomycin (B). Suspension cultures of TA3-Ha mouse mammary tumor cells were treated in vitro with a single dose of macromomycin(B) (MCR) at 1 mug/ml for 24 hours. This dose, which is cytostatic but not lethal, increased the immunogenicity of the cells. Adoptive transfer of spleen cells sensitized to MCR/TA3-Ha cells evoked an immune response against MCR-treated TA3-Ha cells but not against normal TA3-Ha cells. The therapeutic effect of MCR treatment (in this case) was, to a very small extent, due to the cytostatic action and more profoundly to the increased immunogenicity of the cells so treated.


Allogeneic tumor-syngeneic muscle sandwich grafts: a method for promoting nonspecific growth. In H-2-incompatible mice, four strain-specific transplantable murine tumors grew when sandwiched between layers of muscle syngeneic to the intended recipient. The allogeneic tumor-syngeneic muscle sandwich (TMS) graft did not grow in recipients presensitized to the allogeneic tumor and in recipients immunized concurrently with tumor or skin allografts from the same strain as the sandwiched tumor. Third-party muscle, allogeneic to both tumor and host, also was effective in promoting nonspecific growth. The escape of the allogeneic tumor from immunologic destruction when it was implanted as a TMS graft may have been due to an afferent blockade.


Study of prenatal DES exposure and its consequences in Rochester, Minnesota. An epidemiologic study of the effects of prenatal diethylstilbestrol exposure in Rochester, Minnesota, is described. Application of the identification and follow-up techniques to other drugs now used during gestation may aid in the ealier discovery of unexpected adverse effects.


Use of the neonatal mouse in studying long-term effects of early exposure to hormones and other agents. The neonatal female mouse is considered as a model for studying the long-term consequences of exposure of the human fetus and neonate to hormones and other agents. Parallelism is noted between the results of administration of sex steroids and diethylstilbestrol (DES) to newborn mice and the phenomenon of vaginal cancer in young women whose mothers were given DES for threatened abortion. The progression of the neonatally steroid-treated mouse lesions from persistent vaginal cornification through hyperplastic lesions to tumors is described. The interaction of progesterone with estradiol is considered (lesions are fewer but more severe at 12 months of age following neonatal exposure to a combination of estradiol and progesterone), and the ability of neonatal progesterone treatment alone to result in cervicovaginal lesions in intact mice is emphasized. All steroids result in increased mammary tumor incidence and lowered age of tumor onset in intact mice bearing the mammary tumor virus; both the ovary and the virus are required for these effects. Possible ramifications of early perinatal exposure are indicated in regard to the male, to nongenital structures, to the endocrine system generally, and to immunologic mechanisms.


Summary of the changes in the human female genital tract as a consequence of maternal diethylstilbestrol therapy. The current evidence associating maternal ingestion of diethylstilbestrol (DES) with the subsequent appearance of clear-cell adenocarcinoma of the genital tract has been presented. Thus far, these cancers are rare among exposed females. DES, dienestrol, and hexestrol, all nonsteroidal synthetic estrogens, have been implicated. The tumors have appeared primarily in exposed subjects over the age of 14 yr with the median ranging from 18 to 19 yr. Nonneoplastic abnormalities have been observed commonly in the exposed, including vaginal adenosis, cervical erosion (ectropion), and transverse vaginal and cervical ridges. The highest incidence of vaginal adenosis was noted among those whose mothers began DES in early pregnancy, and no cases were observed among those whose mothers began treatment after the 18th wk. Current evidence suggests that these alterations as well as the carcinomas are of mullerian origin.


Circulating levels of prolactin and growth hormone and natural incidence of mammary tumors in mice. Radioimmunoassay measurements of hormones in sera show that contrary to expectations, natural mammary tumorigenesis in mice is not always accompanied by high prolactinemia. Basal levels of circulating PRL follow no consistent pattern in relation to the incidence of mammary tumors. Circulating PRL after a provocative stimulus, however, presents a profile characteristic of the mammary tumor incidence: strains with high incidence of mammary tumors generally have have lower levels, whereas strains with low incidences exhibit higher levels. These differences in circulating PRL levels are apparently not caused by differences in the pituitary stores or pituitary release of PRL. The PRL release patterns appear not to be entirely genetic, since they can be modified by foster nursing, a procedure that may introduce MTV in a virus-free strain. The urine concentrations of PRL and GH and the tissue uptake data suggest that the C3H/St strain may metabolize PRL and GH differently from the C5BL/St strain. It remains to be determined whether the observed differences are significant factors in the development of spontaneous mammary tumors in mice.


Metabolic activation of carcinogenic diethylstilbestrol in rodents and humans. In vivo biotransformation of diethylstilbestrol (DES) was studied by radio-GLC and GLC-mass spectrometry using both radioactively and deuterium-labeled DES. Among the urinary and biliary metabolites identified in intact Wistar rat and Syrian golden hamsters are dienestrol and hydroxy and methoxy derivatives of dienestrol and DES. The identification of 4'-hydroxypropiophenone as a urinary metabolite of DES in the rat is consistent with the hypothesis that dienestrol is formed via an epoxide-diol pathway. Some of the metabolites imply electrophilic reactivity according to their chemical structure and may represent proximate carcinogens of DES. In humans, dienestrol and hydroxy dienestrol constitute the major urinary DES metabolites in men and were also identified in the urine of a woman. Considerable species differences in DES metabolism between humans and rats were found with regard to the route of excretion and the pattern of urinary metabolites.


Testing the antifertility drug enovid for carcinogenesis in five strains of mice. In testing for any carcinogenic effect of certain hormones the choice of test strain or strains is extremely important. By selecting several inbred strains one obtains maximum genetic variability offering a greater chance of demonstrating carcinogenesis in regard to a number of organs or tissues. The five strains of mice chosen in this study (C3H/He, C3HfB/He, BALB/cHe, A/He, and C5BL/He) provided a test for effect of Enovid at sterilizing doses on occurrence of mammary tumors, ovarian tumors, hepatomas, cervical and vaginal tumors, adrenocortical adenomas, and hypophyseal tumors. Results indicated a slight inhibitor effect of Enovid on mammary tumors in C3H and adrenocortical adenomas in BALB/c and an even greater inhibiotry effect on hepatomas in C3HfB. No effect on ovarian tumors could be detected in the appropriate test provided by C3HfB. In BALB/c females epithelial lesions of the cervix and vagina were observed in both the treated females and the controls with a slight increase in the group treated with the highest dose. All were small lesions observed only in the histologic section. None had invaded beyond the wall of the vagina, and none had metastasized. None could be classified as a frank carcinoma. In the group treated with the highest dose of Enovid, the lesions showed slightly further progression than in the other groups. The most significant tumorigenic effect of the Enovid was the increase in hypophyseal tumors in the C5BL females at advanced age. Direct extrapolation from one strain to another, from one species to another, from mouse to humans, or from one human being to another would be risky because of genetic differences. Approaches to the problem in human beings are, however, suggested. Because of these observations in mice, it would be well, if possible, to collect hypophyses of women at postmortem who have previously been on birth control pills for some time and send these hypophyses to some central laboratory where they can be examined for any changes that might be attributed to the use of the antifertility drug.


Dose-dependent kinetics of diethylstilbestrol in the rat. Diethylstilbestrol (DES) acts as a transplacental carcinogen with organotropic properties. The question is raised whether the pharmacokinetics of DES contribute to this effect. [3H] DES was administered to female Wistar rats orally, intravenously, and introperitoneally at doses ranging from 0.025 to 25 mg/kg. Radioactivity associated with erythrocytes and plasma proteins was measured and the pattern of unconjugated metabolites and glucuronides after column and thin-layer chromatography in blood determined. The different routes of administration resulted in different metabolite patterns. After iv injection, unconjugated, unpolar metabolites predomininated; after ip injection, glucuronides of unpolar metabolites; and after oral administration, unconjugated polar metabolites. The different compositions of the glucuronide fractions remained constant with time. The composition of the unconjugated metabolites changed with time after oral administration but not after iv injection. Blood levels of metabolites did not increase proportionally with dose. Overproportional increase was most prominent with the highest dose used. The observed effects are explained with intestinal metabolism and a liver first-pass effect.


Effects of acute anoxia on renal transport processes. Acute disruption of the renal blood supply has been shown by several workers to alter renal cortical metabolic functions and some transport processes. The present study was designed to examine acute ischemic effects on transport functions for both organic and inorganic substances. Acute clamping of the renal artery, renal vein, and ureter for 45 min produces a reversible disruption of tissue electrolyte and water balance. Longer occlusion appeared to produce irreversible effects. Alpha-Aminoisobutyrate (AIB) and lactate-stimulated rho-aminohippurate (PAH) transport were altered selectively by the 45-min occlusion. A longer occlusion period also depressed base-line PAH and tetraethylammonium (TEA) transport. Some of the depression of the organic compound transport functions are reversible. Renal cortical oxygen consumption measured in vitro was affected minimally.


4-aminobiphenyl, 2-naphthylamine, and analogs: analytical properties and trace analysis in five substrates. Methods for monitoring trace levels of 4-aminobiphenyl, 2-naphthylamine, and their hydrochloride salts in waste water, microbiological growth media, potable water, human urine, and mouse blood utilizing spectrophotofluorometry (SPF) are described. The salient elements of the methods are extraction of the residues as the free amine with benzene, rapid cleanup on an alumina column, and quantification of the free amine in methanol via SPF. Potable water solutions of the salts are diluted with 0.01 N aqueous HCL and quantified directly by SPF. Ancillary analytical information concerning gas chromatography of the free amines, partitioning properties of the compounds between solvent pairs, their solubility and stability in water, and thin-layer chromatographic data is presented. The compositions of various admixtures of 1- and 2-naphthylamine or their salts were determined by using SPF with calculations based on simultaneous equations.


Tissue distribution of (14C) methyl mercury in the lobster, Homarus americanus. [14C] Methyl mercury was administered by three different routes: intravascular (iv) injection, ingestion, and absorption from the ambient water. After iv administration (0.1 mg/kg) [14C] methyl mercury was rapidly removed from the plasma, followed by slow loss from the hepatopancreas and a strikingly persistent increase in the amount of radioactivity in the tail muscle. Most (80-90%) of the radioactivity in the hepatopancreas was shown by TLC methods to be the parent compound, and approximately 10% of this persisted for 6 days after injection. The half-life in this organ was found to be 21 days. One month after iv treatment with methyl mercury, the only organs that contained more than 0.1 ppm of this xenobiotic were egg masses, male gonads, heart, brain, intestine, and tail muscle. The half-lives for disappearance from sexual organs were greater than 1 month. After ingestion of [14C] methyl mercury (0.1 mg/kg) in food the hepatopancreas contained most of the administered dose at 6 days (68%), while the stomach (10%), tail muscle (8%), and carcass (15%) contained less. A unique distribution pattern emerged 6 days after exposure to [14C] methyl mercury-containing ambient water (0.1 ppm). The tail muscle contained most (50%) of the absorbed dose, whereas the hepatopancreas and carcass contained only 23 and 10%, respectively. In view of the small molecular size and high lipid solubility of methyl mercury and the lipophilic properties of the chitin-protein exoskeleton of the lobster, it is likely that significant uptake directly from the water as well as storage of absorbed methyl mercury occurred in the tail region. Residue analysis on untreated lobsters indicated that the egg masses contained the largest amount of methyl mercury (0.1 ppm). The hepatopancreas and carcass (muscle) levels were less than 0.05 ppm.


Effect of water and food deprivation on hepatic microsomal metabolism of hexobarbital and aniline. Water deprivation for 48 hr with its accompanying decrease in food intake significantly lowered the in vitro rate of hexobarbital metabolism by hepatic microsomes isolated from male rats. Pair-fed rats allowed water ad libitum had a significantly lower level of hexobarbital metabolism than those deprived of water. Rats starved for 24 hr with or without water also had levels of hexobarbital metabolism significantly lower than their controls; with those animals allowed water ad libitum, the level was significantly lower than for those deprived of water. In vivo hexobarbital "sleeping time" experiments were in general agreement with these results. The in vitro metabolism of aniline was increased in both male and female rats following 24 hr starvation and in female rats (but not males) the effect was greater when water was allowed than when deprived. The differences between hydrated and dehydrated animals were not attributable to reduction in concentration of microsomal protein or the water content of liver. It is concluded that water consumption accentuates the effect of food deprivation on hepatic microsomal metabolism.


Cadmium in liver, kidney, and hair of humans, fetal through old age. Cadmium concentrations were measured in the liver, kidney, and hair of 107 individuals from the Cincinnati area. The tissue samples were obtained from hospital and coroner autopsy cases and represent a cross-section of individuals from fetal to old age. The sections used were standardized and analyzed for cadmium by atomic absorption. Information obtained at the time of autopsy included the cause of death, the conditions of the tissues, the body height and weight, the amount of anthracosis in the lungs, as well as age, sex, and race. The liver concentrations increased steadily until the late decades. The kidney concentrations increased to the fourth decade, peaked, and decreased steadily from the sixth decade thereafter. The leveling effect in the liver may have been due to the increased presence of fatty livers, and the kidney decreases to nephrosclerosis. The kidney and liver were statistically well correlated. Hair, which achieved a surprisingly high Cd concentration in early infancy, tended to decrease throughout the rest of the age span and correlated poorly with the liver and kidney. There seemed to be no relationship between the amount of pulmonary anthracosis and the cadmium found in any of the tissues. The results obtained here for liver and kidney were in line with other reports in the literature; however, the hair results were somewhat different.


Heptachlor: uptake, depuration, retention, and metabolism by spot, Leiostomus xanthurus. The estuarine fish, spot (Leiostomus xanthurus), was exposed to 0.27, 0.52, 1.01, 1.99, and 3.87 mug/liter technical grade heptachlor (65% heptachlor, 22% trans-chlordane, 2% cis-chlordane, 2% nonachlor, and 9% unidentified compounds) for 24 days in a flowthrough bioassay, followed by 28 days in heptachlor-free seawater. Concentrations of heptachlor, heptachlor epoxide, and trans- and cis-chlordane in edible tissues were monitored at day 3 and weekly thereafter throughout the bioassay and at the end of the postexposure period. All four chemicals were accumulated by spot. Maximum concentrations of heptachlor were observed on day 3; maximum concentrations of the other three compounds were observed on day 17. The average bioconcentration factors for heptachlor and trans-chlordane were 3,600 and 4,600, respectively. Only 10% or less of the maximum concentrations of heptachlor, heptachlor epoxide, and trans-chlordane accumulated during the exposure period remained after 28 days in pesticide-free seawater; an average of 35% of the cis-chlordane remained. Relative total amounts of heptachlor and cis-chlordane changed during the exposure and post-exposure periods. Nearly all of the heptachlor was eliminated or metabolized to its epoxide. Cis-chlordane, which averaged 4-7% of the total residues (chlordanes and heptachlors) in edible tissues during the exposure, increased to 18-23% of the total residues by the end of the postexposure period.


Aspartame--a sweet surprise.

Aspartame--a sweet surprise. The dipeptide ester L-aspartyl-L-phenylalanine methyl ester (APM) has been found to have a remarkably clean, sucrose-like taste with no off flavor and a potency 150-200 times sucrose. Subsequent work has shown that many alpha-amides of L-aspartic acid are sweet. Some results of stability studies and a taste panel evaluation of APM are reported.


Effect of lead on glycine cleavage activity in rat liver mitochondria. Change of glycine cleavage activity in liver mitochondria of lead-intoxicated rats was studied in this paper. The following results were obtained: (1) The activity in hepatic mitochondria of rats was decreased by a single administration of lead (4 mg/100 g body weight); (2) in vitro the activity in the normal rat liver mitochondria was markedly inhibited by addition of lead acetate (4mM).


Toxicological evaluation and pharmacokinetic profile of beta-hydroxypropionitrile in rats. Beta-hydroxypropionitrile (beta-HPN) was incorporated in the drinking water of rats in order to define the toxicological effects associated with repeated daily exposure to this compound. Parameters evaluated were appearance and demeanor, mortality, body weight, food consumption, water consumption, hematology, terminal organ weights and organ to body weight ratio, and gross and microscopic pathology of tissues. No untoward effects were associated with the ingestion of beta-HPN at the dose levels utilized.


Biliary excretion of cadmium by rats: effects of zinc, cadmium, and selenium pretreatments. The significance of bile as an excretory route for cadmium (Cd) was studied in anesthetized, bile-duct-cannulated Sprague-Dawley rats during a 6-hr collection period. Observations were made on bile flow rates, the concentrations of Cd in bile following dietary and parenteral Cd exposure, and the influences of zinc (Zn), selenium (Se), and Cd pretreatments upon the biliary excretion of subsequently administered Cd. The bile flow rates ranged between 1.96 and 2.89 mg/g rat-hr (22 +/- 3 ppb Cd) for normal rats and between 2.68 and 4.09 mg/g rat-hr (58 +/- 6 ppb Cd) for rats fed 100 ppm Cd. Less than 0.1% of the Cd administered subcutaneously at rates ranging from 0.25 to 40 mg/kg rat could be accounted for in bile collected during the 5-hr period following the parenteral Cd injections. Subcutaneous administration of 8 mg Zn/kg rat or 0.5 mg Cd/kg rat on days 1 and 6, respectively, before the postcannulation administration of 1 mg Cd/kg rat caused a significant reduction in the biliary excretion of Cd during the bile collection period. Administering 2 mg Se/kg rat 3 days prior to the postcannulation administration of 1 mg Cd/kg rat caused a significant increase in the biliary excretion of Cd during the bile collection period. The biochemical bases for these observations are believed associated with the type of metal-binding protein induced by the respective pretreatments.


Effects of piperonyl butoxide on dimethylnitrosamine metabolism and toxicity in Swiss mice. The effects of piperonyl butoxide (PB) on the metabolism and toxicity of dimethylnitrosamine (DMN) in Swiss mice were determined. PB, at doses of 10 and 20 mg/kg, inhibited DMN demethylase 45 min after treatment by 18 and 37%. These inhibitory effects were marked 0.5 hr after PB (640 mg/kg) treatment and reached maximal effects at 2 hr when there was 55% inhibition of enzyme activity. The inhibition by PB continued for 24 hr where enzyme activity was suppressed by 35%. At 48 hr after treatment there was stimulation of enzyme activity. Enzyme kinetic determinations showed no change in Km but Vmax decreased from 129 to 49 mumol CH2O/min-g liver. PB (640 mg/kg) inhibited DMN (500 mg/kg; im) mutagenicity in the host-mediated assay, decreasing the mutant frequency by 42%. Paradoxically, PB (640 mg/kg) had no effect on the alkylation of nucleic acids or proteins in mouse liver, kidney, lung, or spleen. In addition, pretreatment with PB (640 mg/kg) had no effect on the LD50 of DMN.


Cellular and subcellular distribution of the S-100 protein in rabbit and rat central nervous system. The cellular and subcellular distribution of the S-100 protein in rabbit and rat central nervous system was studied both quantitatively and qualitatively. Microcomplement fixation estimations on bulk-prepared neuronal and glial cells showed at least five to six times higher amounts of water-soluble S-100 in glial cell-enriched fractions as compared to fractions enriched in neuronal perikarya. Nerve cell fractions contained a higher percent of tissue-bound S-100 protein. High levels of S-100 protein were found in mitochondria and soluble protein fractions. Immunoelectron microscope investigations demonstrated S-100 protein in neuronal structures, such as the postsynaptic membrane and part of the plasma membrane. Among glial cells the astrocytic filaments contained high levels of S-100 protein. S-100 was also found in most subcellular membranes of astrocytes and oligodendrocytes.


Biosynthesis of DNA and RNA in neuronal and glial cells from various regions of developing rat brain. Slices of cerebral hemispheres, brain stem, and cerebellum from rats 5-30 days old were used for in vitro incorporation of [methyl-3H]thymidine and [6-14C]orotic acid into DNA and RNA, respectively. The rates of DNA and RNA synthesis decreased markedly during development, with the most marked decrease observed for DNA. The different brain regions showed specific patterns of decline of DNA and RNA synthesis. Following incubation of slices, the tissues were fractionated to obtain fractions enriched in neuronal cells and in glial cells. In cerebellum, the granule neurons were separated from the Purkinje neurons. The glial:neuronal ratio of DNA specific activity was different in the three regions examined: in cortex it decreased from 6 at 10 days to 3 at 20-30 days; in brain stem it was 3 throughout 10-30 days; in the cerebellum (glia:granule neuron ratio) it was also 3 at 30 days but only 0.3 at 10 days. Concerning the RNA incorporation, small differences were found between neuronal and glial cells.


Mesencephalic lesions resulting in normophagia, reduced weight and altered metabolism. The effects of bilateral lesions of the ventral noradrenergic bundle (VNA) were studied in male rats. In contrast to data reported by others, hyperphagia and obesity were not observed following VNA lesions. Indeed, except for a depression during the first three postoperative days, food intake (FI) of the VNA lesioned animals (VNAL) was normal. Interestingly, the body weight (BW) of the VNAL was significantly reduced compared to the controls, and a pair feeding study indicated that this depression of BW was not due to their FI. Computation of FI per metabolic size showed that the VNAL actually had a significantly increased FI compared to the controls. After a two day fast the VNAL lost more metabolic size than controls and upon refeeding they defended their pre-fast BW. The VNAL rats showed normal body composition and circulating glucose, insulin and prolactin. They had reduced free fatty acids, triglycerides, growth hormone and body length. The data suggest that the mesencephalon influences BW set point, some metabolites and possibly overall metabolism.


Molecular information structures in the brain. This paper presents a theory of memory and memory mediated learning based on the manipulation of macromolecular conformations. The main features of the theory are: 1) the brain contains primary and reference neurons; 2) inputs from the external environment produce particular patterns of primary firing; 3) the firing of a primary neuron sensitizes certain of its dendrites; 4) the sensitized primaries are loaded by the reference neuron active at the time and in such a way that they fire when called by this reference neuron, thus reconstructing the original pattern of primary activity. The reference neurons may also be loaded by primaries, thus making it possible for the reconstruction process to be initiated by some feature of the initial input. Each reference neuron loads and calls at most one primary pattern of activity, thereby preventing superposition of memories. If the primaries are loadable by sequences of impulses, this makes it possible to increase the connectivity among the various types of neurons by using party-line organization. The loading and calling processes themselves are mediated by call molecules. These are allosteric enzymes, located in the dendrites of primary and reference neurons, whose states are set either by an impulse or sequence of impulses and which catalyze events leading to impulse formation whenever this input recurs. The call molecules are capable of duplicating their setting (or conformation) using either intra- or interneuronal potentials, thereby ensuring stability of the memory trace. The theory allows for general powers of memory manipulation (by rememorization), for the construction of time ordered, content ordered, and associative data structures, and for computation with global representations of the environment. It makes a large number of testable predictions, provides a natural interpretation for the structure of the cerebral cortex, and accounts for: resistance to cooling, differential effects of chemical agents on short and long term memory, distributed character of memory, accessibility of memories by stimulation of specific brain loci, and also the details of classical conditioning and instrumental learning.


Checklist for a children's trauma room.

Checklist for a children's trauma room. Optimal management of a severely injured child depends upon instant availability of a wide variety of emergency equipment and supplies. Some of these items are not commonly found in trauma rooms designed for adults, and others are required in a range of sizes unique to child care. After a major resuscitative effort the trauma room is a shambles from which it must be restored rapidly to receive the next trauma victim. A checklist has proven invaluable in preventing disastrous omissions when the trauma room is being restocked. This list was patterned after a similar checklist used in the Emergency Department of the Parkland Hospital in Dallas and was developed for children by the Trauma Committee of the American Pediatric Surgical Association. It has been tested and refined in the Trauma Room of the Children's Hospital, Columbus, Ohio, during a 2-year period in which 100,000 children presented to the emergency department. It should be helpful in any emergency department which receives seriously injured children.


Arterial injuries of the extremities.

Arterial injuries of the extremities. Fifty-eight patients with arterial injuries of the extremities were treated during the past 8 years. Fifty-one had acute injuries and seven had nonacute injuries. Blunt trauma or shotgun wounds caused 74% of the injuries, and 55% were associated with skeletal trauma. All the acute injuries endangered the limb; the average ischemic time was 8.5 hours. Brachial, popliteal, and superficial femoral arterial injuries were seen most frequently. Repair was accomplished with autogenous saphenous vein grafts in 47% and end-to-end anastomosis in 41%. Six patients died, four due to injuries of other organs. The most alarming complication of arterial repair was secondary hemorrhage which occurred in three patients and was caused by local infection. Six patients (13.3% required amputations; the highest number (three) after injuries of the popliteal artery. The injuries leading to amputations had associated prolonged ischemia, severity of injury, and associated venous, soft tissue, and skeletal injury. The nonacute injuries were in the form of false aneurysms, pulsating hematomas, AV fistula, and delayed bleeding. These were easily managed without any significant complication.


Fractures of the mandible in children.

Fractures of the mandible in children. A review of 219 fractures of the mandible in children treated from 1960 through 1970 has been presented. The etiology, location, and treatment required were considered in relation to patient's age and dentition. The most common form of treatment was closed reduction and intermaxillary fixation.


Aspiration pneumonitis and pulmonary phospholipids. Peptic aspiration pneumonitis (Mendelson's syndrome) results when gastric acid is aspirated into the lung, as may occur during anesthesia. In the present study, 0.1 N HCl was insufflated via the endotracheal tube into pentobarbital-anesthetized dogs in an amount sufficient to cause severe pulmonary damage. At death, the thorax was opened, the lungs grossly examined, and either weighed and desiccated for determination of wet/dry lung weight ratios, rinsed with saline for removal of alveolar surface phospholipids, or homogenized for whole lung phospholipid determination. Gross appearance and wet/dry lung weight ratios indicated severe pulmonary edema. The surface tension values of the lung wash were elevated over control values. Lysophophatidyl-choline (LPC) showed a striking increase over control values. Because LPC is a potent hemolytic agent which builds up in the lung following this pulmonary insult, and because increased hemorrhaging gradually develops following experimental acid insufflation, it is concluded that LPC is most probably causally related to the hemorrhagic pneumonia of Mendelson's syndrome.


Effects of albumin and/or furosemide therapy on pulmonary edema induced by hydrochloric acid aspiration in rabbits. Aspiration of hydrochloric acid in rabbits resulted in an increased P(A-a)O2 together with increases in both lung water volume and lung extravascular albumin. This finding suggests lung damage following acid aspiration is related to changes in capillary permeability, with pulmonary edema resulting from the movement of albumin and water into the interstitial space. Therapy with albumin and furosemide together reduced the lung water and albumin accumulation and decreased P(A-a)O2. Treatment with albumin or furosemide alone was ineffective. Caution should be exercised in administering albumin alone for therapy of pulmonary edema when plasma protein is not clearly decreased, or when increased pulmonary capillary permeability is suspected.


Abdominal stab wounds: evaluation of sinography. This paper reviews a 30-month experience with 172 patients suffering abdominal stab wounds treated at the Denver General Hospital, during a period when policy included liberal sinography and all penetrating injuries were explored. Laparotomy was performed in 87%. Of the 65 patients undergoing sinography, 62% indicated peritoneal penetration; of these, 30% had no visceral injury. An additional 10%, with minor intraperitoneal injuries, probably would not have required celiotomy. In 25 cases the stab penetrated the peritoneal cavity after first entering the chest. Peritoneal tap and peritoneal lavage were used in 10 patients. It is concluded that the cost/benefit ratio of sinography is so poor that it is rarely indicated. When doubt exists as to significant intraperitoneal pathology following an abdominal stab wound, close observation without sinography is recommended for determining indication for laparotomy.


Traumatic tattooing.

Traumatic tattooing. Traumatic tattoos may be classified into abrasive and explosive types. The abrasive form leaves the pigment deposit in the more superficial layers of the skin. Explosive forms of traumatic tattooing usually have a central focus of deeply embedded detritus with more superficially placed particles radiating from the central focus. In either form, the best results are achieved by immediate meticulous care of the wound. Many instruments and abrasive devices have been used to remove the embedded pigmented debris, yet the stimplest and most successful is the sterile, hard, natural-bristle toothbrush. This can easily be sterilized, packaged, and kept available in the emergency room and in the operating room.


Gunshot wounds of the extremities in children. A ten-year review of gunshot trauma in Manitoba was prompted by an increasing incidence in gunshot trauma in children. Of 54 children treated from 1964 through 1973 60% sustained injury to one or more extremities. Fifteen children sustained long bone fractures, usually complete and often comminuted. All fractures united and there was only one instance of osteomyelitis. The major complication in gunshot wounds to the skeletally immature was limb length inequality secondary to epiphyseal plate injuries. This complication occurred in nine children. In three children there was in epiphyseal growth disturbance, even though the missile did not appear to pass through the plate but only in close proximity to it. Joint contractures secondary to skin loss and resulting subsequent skin contracture were also common. The amount of debridement required following gunshot trauma in children appears to be somewhat less than is recommended for similar injuries sustained by adults in time of war.


Bronchial cytologic changes during cardiopulmonary bypass: iron transport by histiocytes in low flow states. We previously reported (1) that tracheobronchial secretions of patients in hemorrhagic shock contained significantly more iron laden histiocytes than secretions of matched controls and demonstrated by histiologic studies in rats bled to hemorrhagic shock that these animals' lungs contained significantly more iron laden histiocytes than those of a control group. Two explanations appeared plausible: 1) abnormal hemoglobin breakdown products of systemic hypoperfusion during shock (2, 3, 5) may have accumulated in the pulmonary circulation where they were phagocytozed by pulmonary histiocytes, or 2) circulating iron laden histiocytes may have been arrested in the lung, which is well known to act as a filter during low-flow states (4). The present study reports on assay of bronchial secretions of 12 open-heart surgery patients under cardiopulmonary bypass from specimens obtained 1) immediately after onset of anesthesia; 2) during; and 3) after termination of cardiopulmonary bypass. The percentage of iron laden histiocytes rose from 15.6+/-6.0 after intubation, to 49.6+/-6.8 after 1 hour on bypass with occluded pulmonary arteries (p less than 0.0025), remaining close to the latter value after pulmonary circulation was restored. Hypoperfusion, accumulation of metabolites, or falling pH at the tissue level are probable explanations for these findings.


Gentamicin dosage in children with extensive burns. During the treatment of Gram-negative bacillary infections in severely burned children, it was noted that administration of standard dosages of gentamicin commonly produced inadequately low serum concentrations of the antibiotic. Pharmakokinetic studies demonstrated that mean peak serum gentamicin levels were significantly lower in 18 burned children as compared with peak serum levels in five unburned children. Furthermore, inadequately low peak serum levels (less than 3.0 mug/ml) were observed in 15 of 18 burned children and in only one unburned child. Clearance studies in two children suggested that the increased requirement for gentamicin can result from either loss of the antibiotic across burn wounds or increased urinary excretion. These studies indicate that the therapy of severe infections in children with major burns often requires administration of gentamicin at higher doses and more frequent intervals than usually employed.


Increased dosage requirements of gentamicin in burn patients. In 14 burn patients treated for serious Gram-negative infections, the use of the previously recommended gentamicin dose of 5 mg/kg/day was found to result in subtherapeutic serum concentrations (peak concentration less than 4 mg/L). The gentamicin half-life was found to be unusually short especially in the younger burn patients. Because of this shorter half-life the dosage interval was decreased to 4 hours to prevent extended periods of subtherapeutic serum concentrations. In addition, the daily dose of gentamicin was increased to achieve therapeutic peak concentration. Individualizing each patient's gentamicin regimen was thought to be instrumental in the favorable response of two patients with Pseudomonas ecthyma gangrenosum. The results of this study would strongly support the measurement of serum gentamicin levels in all burn patients with life-threatening infection. The gentamicin dosage regimen should then be individualized for each patient to provide optimal peak concentrations. In addition, patients demonstrating a short drug half-life may require a decreased dosage interval to prevent prolonged periods of sub-therapeutic concentrations.


Occult traumatic dislocation of the patella. In series of patients with clinically established traumatic patellar dislocation other authors have found surgical and radiological lesions pathognomonic for this condition. In a series of patients admitted with hemarthrosis, and with no knowledge of having had dislocation of their patella, we found the same lesions thus illustrating that a syndrome of occult traumatic patellar dislocation exists. The vertical location of the patients' patellae was found normal measured by the index of Insall and Salvati (9). The intercondylar groove measured by the method of Brattström (4) was found in almost all cases to be under the upper normal limit. Their knee caps were normal following the descriptions of Knutsson. None had valgus knees beyond normal upper limits and none had general laxity of joints, thus indicating that this syndrome may be found in normal knees.


Dynamic fixation of the avulsed clavicle. A new operative approach is presented for treatment of the fractured distal end of the clavicle, which is associated with a disruption of the coracoclavicular ligaments. Three cases are presented in which open reduction and the clayicle held in the corrected position by transfer of the coracoid process with its attached muscles onto the clavicle was followed by complete recovery.


Treatment of traumatic myositis ossificans circumscripta; use of aspiration and steroids. The classification, clinical presentation, forms of treatment, and theories of pathogenesis of traumatic myositis ossificans circumscripta are reviewed. A group of 7 patients with this lesion was treated with hematoma aspiration and injection of steroids, lysosomal enzymes, and xylocaine, resulting in significant limitation of disability. The determining factor in the development of this disorder seems to be the response of local soft tissues to trauma rather than the magnitude of the trauma itself which we feel accounts for the success of the form of treatment described.


Management of acute laryngeal injury: a critical review. The approach to management of 30 cases of laryngeal trauma coming to open exploration is outlined. Eighteen patients had blunt trauma and 12 had penetrating wounds. Four cases resulted in airway compromise and all were in the blunt trauma group. Although significant granulation tissue response occurred in only eight of 30 cases, it was present in three of the four concluding with stenosis. It is demonstrated that the use of a stent, with a tissue graft, when mucosal approximation cannot be accomplished, facilitates prevention of endolaryngeal distortion and webbing and maintenance of skeletal framework integrity; its use did not predispose to granulation tissue development. A principal etiologic factor in those patients who had good airway but only a fair voice was arytenoid fixation, this despite anatomic reduction when subluxation occurred. Recurrent nerve injury was identified in only one patient. Foremost in management is suspicion and recognition of laryngeal injury in the multiple trauma patient. Once the urgent problems of hemorrhage, shock, and airway are attended to, attention must be directed to a systematic evaluation of the neck and larynx.


Special problems of vascular injuries in children. Vascular injuries in children are not rare. The rising incidence of general trauma in children and the frequent use of diagnostic procedures in this age group may contribute to an overall increase in these injuries. A 10-year experience with severe vascular injuries in children at The Johns Hopkins Hospital has been reviewed. The mechanisms of production may be similar to those in adults, but many injuries are unique to the pediatric age group. The sequelae of such injuries in infants and children may include abnormal growth and development of the involved extremity. Complications and special surgical management of our group of 71 children have been discussed and preventive measures have been suggested.


Penetrating stab wounds of the chest: experience with 200 consecutive cases. The incidence of penetrating wounds of the chest is rising rapidly. Opinions continue to differ on their management. Our experience with 200 consecutive cases of stab wounds of the chest between 1972 and 1975 were reviewed. There were 176 males and 24 females. The average age was 31 years; about two-thirds of the patients were under 30. About 74% presented with hemothorax or hemopneumothorax; 21 presented with pneumothorax. Eleven per cent had associated intra-abdominal injuries. Seventy-nine per cent were successfully treated with tube thoracostomy. About 15% underwent thoracotomy, with three deaths (mortality, 10%); the mortality for cardiac wounds was 16%; overall mortality was 1.5%. The overall complication rate was 5.5%, most occurring in patients with cardiac wounds and associated intra-abdominal injuries. The average period of hospitalization was 6.5 days. Treatment was individualized. Indications for each course of therapy are discussed.


Puncture wound of the eyelid causing brain abscess. A case of brain abscess following an apparently trival puncture wound to the eyelid is presented to emphasize the importance of meticulous examination of eye wounds in children to look for penetration into the cranial vault. Any suggestion of such penetrating injury either by history, examination, or X-ray should dictate neurosurgical consultation, and immediate local culture and debridement of the wound. If nonoperative management is elected, a careful watch must be kept for signs of infection, which dictate early exploration, and broad-spectrum antibiotic coverage of Gram-positive and Gram-negative organisms common to penetrating skull trauma.


Bilateral sacroiliac joint dislocation with intrapelvic intrusion of the intact lumbosacral spine and sacrum. Two cases of bilateral sacroiliac joint dislocation with intrapelvic intrusion of the intact lumbosacral spine and sacrum are reported. This condition is a distinct traumatic entity. The essential features of this condition are the disruption of the pelvic ring with bilateral sacroiliac dislocation. The spine as a whole descends into the depths of the pelvis. Disruption of the anterior structures but not always the pubic symphysis is apparently a prerequisite for the development of the condition. Extensive soft-tissue damage is part of the injury. The etiology is a direct crushing violence to a standing or walking patient. It is assumed that this injury is less rare than it seems, and that the main reason for not being mentioned is the rarity of the survival of the patients. As a result of success in the intensive care of the severly injured, more survivals and more reports are anticipated.


Sensorineural hearing after removal of cholesteatoma from labyrinthine fistulae. An investigation was undertaken to determine what factors improve or worsen bone conduction thresholds following treatment of labyrinthine fistula produced by cholesteatoma. It is suggested that closure of the fistula and reconstruction of normal sound transmitting pathways by ossicular reconstruction produce a favorable mechanical situation to produce this response. In addition there remains the possibility that removal of cholesteatoma matrix may prevent a toxic degeneration of the cochlea at the site of the fistula. Therapy advocated is a two stage combined approach tympanoplasty.


[Tympano-ossicular homografts].

[Tympano-ossicular homografts]. Following a few European and American authors, we have undertaken tympano-ossicular homografts at the Hotel-Dieu de Quebec. The organization of the bank was achieved with many difficulties. We undertook our first surgical homografts in 1974, and 27 cases have since been performed. Our statistics are based on 19 cases. We obtained a perfect tympanic membrane in 17 cases, and 60 per cent of these have shown good improvement in hearing. This percentage is remarkable when one considers that we limited this procedure to cases for which all other techniques were bound to fail.


Round window fistula.

Round window fistula. Round window fistula as a cause of sudden hearing loss was diagnosed in five people. It was associated with a sudden change in middle ear pressure in three, with heavy lifing in one, and was probably caused by a significant blast exposure in the other. The clinical features of the condition are varied, with hearing loss and tinnitus being the only constant findings in our patients. One case demonstrates, perhaps for the first time, the association of an abnormally mobile portion of the round window membrane with vertigo and nystagmus. Suggestions on identification and management of round window fistula are made.


Replica cytology in cancer of the larynx: an evaluation of a replica method in the diagnosis of laryngeal malignancy. A series of 38 cytological samples from the larynx, obtained with a replica method, is presented. The method is described in detail. Conclusive material was obtained in 34 cases. Fourteen per cent false negative evaluations were found. However, when more strict criteria were used for accepting the cytological specimen as suitable for evaluation, no false negative was encountered. It is therefore concluded that this replica method for obtaining cytological samples from the larynx could become a valuable tool as a screening method during indirect laryngoscopy in search of laryngeal malignancy provided very strict criteria are used for accepting the cytological specimen as suitable for evaluation.


Liposarcoma of the tongue.

Liposarcoma of the tongue. Liposarcoma is a rare tumor in the head and neck. No previous report of its occurrence in the tongue is found at the time of this writing and this prompts its presentation. Significant in its management is wide surgical excision. Irradiation may be effective in some metastases. Prognosis is best in well differentiated forms.


[Radiotherapy following surgery of the larynx]. A method of radiotherapy following total or partial surgery of the larynx is described. Criteria for the selection of patients and a technique of treatment are suggested. The results from 1966 to 1971 are reported.


The high jugular bulb.

The high jugular bulb. Three cases of high jugular bulb are presented. On of these occurred as a mass in the external auditory canal simulating an osteoma, and to our knowledge is the first reported case of a high jugular bulb presenting in the external canal. The clinical manifestations are described, and the value of polytomography as an aid to diagnosis is stressed.


Formation of a Sindbis virus nonstructural protein and its relation of 42S mRNA function. Chicken embryo fibroblasts infected with an RNA- temperature-sensitive mutant (ts24) of Sindbis virus accumulated a large-molecular-weight protein (p200) when cells were shifted from the permissive to nonpermissive temperature. Appearance of p200 was accompanied by a decrease in the synthesis of viral structural proteins, but [35S]methionine tryptic peptides from p200 were different from those derived from a 140,000-molecular-weight polypeptide that contains the amino acid sequences of viral structural proteins. Among three other RNA- ts mutants that were tested for p200 formation, only one (ts21) produced this protein. The accumulation of p200 in ts24- and ts21-infected cells could be correlated with a shift in the formation of 42S and 26S viral RNA that led to an increase in the relative amounts of 42S RNA. These data indicate that p200 is translated from the nonstructural genes of the virion 42S RNA and further suggest that this RNA does not function effectively in vivo as an mRNA for the Sindbis virus structural proteins.


Infectious, linear, unintegrated DNA of Moloney murine leukemia virus. A closed circular, double-stranded infectious DNA of Moloney leukemia virus has been described previously. The present report characterizes a second type of infectious, unintegrated viral DNA which is linear, largely double stranded, and of mass comparable to that of the closed circular viral DNA. The linear form is of nonpermuted sequence, and SalI endonuclease cleaves at one site 45% from one end.


Analysis of the envelope of Rauscher murine oncornavirus: in vitro labeling of glycopeptides. The identity and localization of the oligosaccharides of Rauscher murine type C viral glycoproteins have been examined by techniques of in vitro labeling. Terminal sialic acid was labeled with tritium by borohydride reduction after selective periodate oxidation, and galactose was labeled by borohydride reduction after specific enzymatic oxidation of the nonreducing terminal of the sugar. The results were compared with those of protein surface labeling with pyridoxal phosphate or lactoperoxidase catalyzed radioiodination. Examination of the labeled reaction products by polyacrylamide gel electrophoresis in sodium dodecyl sulfate showed that in every case the major component labeled was a glycoprotein of about 70,000 daltons. The identity of this glycoprotein as the virion envelope component was confirmed by immunoprecipitation with mono-specific antiserum prepared against purified Rauscher virus glycopeptides of 69,000 and 71,000 daltons. No other protein or glycoprotein on the surface of the virion was detected, and disruption of virions-before labeling did not reveal additional distinctive glycoproteins. There was minor labeling of sugar residues of other components, but these remain to be characterized and are not now identified as other viral proteins. Studies of the structural organization of virion proteins using the cross-linking reagent methyl-4-mercaptobutyrimidate showed only linkage of the virion envelope or core proteins to themselves. These results indicate that most, if not all, of the oligosaccharides at the surface of Rauscher virus are entities of the 69,000- and 71,000-dalton glycopeptides and that they contain a terminal sialic acid and galactose and a subterminal galactose.


Segmented genome and nucleocapsid of La Crosse virus. La Crosse (LAC) virions purified by velocity and equilibrium gradient centrifugation contained three single-stranded RNA species. The three segments had sedimentation coefficients of 31S, 25S, and 12S by sodium dodecyl sulfate-sucrose gradient centrifugation. By comparison with other viral and cellular RNA species, the LAC viral RNAs had molecular weights of 2.9 x 10(6), 1.8 x 10(6), and 0.4 x 10(6). Phenol-sodium dodecyl sulfate-extracted LAC virion RNA was not infectious for BHK-21 cell cultures under conditions in which Sindbis viral RNA was infectious. Treatment of LAC virus with the nonionic detergent Triton X-100 and salt released three nucleocapsid structures, each containing one species of virion RNA. The nucleocapsids had sedimenation coefficients of 115S, 90S, and 65S. Negative-contrast electron microscopy of the nucleocapsids indicated that they were convoluted, supercoiled, and apparently circular. They had a mean diameter of 10 to 12 nm and modal lengths of 200, 510, and 700 nm (some were even longer). By chemical and enzymatic analysis of purified viral RNA, one type of 5' nucleotide (pppAp) present in the proportion of one per RNA segment was identified. After periodate oxidation, each virion RNA species was labeled by reduction with [3H]sodium borohydride. Taken together, these results suggest that although the nucleocapsids appear as closed loops, the viral RNA has free 5' and 3' ends and is, therefore, not circular.


Purification and composition of the proteins from Sindbis virus grown in chick and BHK cells. Procedures are described for the purification of the Sindbis virus structural proteins. The amino acid and carbohydrate compositions of the purified proteins are presented for virus grown in BHK-21/13 and chicken embryo cells. Glycoprotein E1 from virus grown in BHK cells is deficient in a mannose-rich glycopeptide found on that glycoprotein when virus is grown in chicken embryo cells. The complex glactose-containing glycopeptides appear similar for virus grown in both hosts. However, when virus is grown in BHK cells, both glycoproteins are enriched in those glycopeptides containing more sialic acid. Since the two viral glycoproteins are difficult to separate cleanly during purification, it is suggested that there may be strong, but noncovalent, interactions between glycoproteins E1 and E2. It is also suggested that there may be an interaction between glycoprotein E2 and a component of the nucleocapsid.


L cell virus: infectivity for type N and B mouse embryo cells. The efficiency of focus formation of four L cell virus populations was significantly higher on type N mouse embryo fibroblast cultures than on type B cells. [5-3H]uridine-incorporating material sedimenting at a density of 1.158/g/cm3 was observed in the supernatant fluids of type N and B cultures infected with the four viral populations.


Clinical analysis of angina pectoris and angina-like pain --With special reference to ECG during attack, "cervical spondylosis" and selective coronary arteriography. In a series of 63 patients, 60 with angina pectoris and 3 with cervical spondylosis and "thoracic spondylosis" showing angina like pain detailed assessments were made of the mode of onset of attack, including electrocardiography during attacks, X-ray examination of the thoracic and cervical vertebrae and neurological examinations, along with coronary arteriography in some cases, with the following results: 1. The cases of angina pectoris were classifiable grossly into two groups according to mode of onset of chest pain: Group A: Angina began with pain in the anterior chest (39 cases); Group B: Angina in the anterior chest was preceded by "pain" occurred elsewhere in the chest (21 cases). The cases in group B were further classified under two categories, types BI and BII, the former being characterized by a sudden onset of "pain" in a somatic area or areas other than the anterior chest where there is usually no dysesthesia, followed by development of retrosternal or precordial pain (6 cases), while the latter type of angina began with paroxysmal exacerbation of preexistent dysesthesia in the nape, shoulder and arms and eventuated in pain in the anterior chest (15 cases). There were two subtypes in the type BII angina viz. types BIIa and BIIb. There was no ECG evidence of ischemic changes at exacerbation of the nucha-omo-brachial dysesthesia in type BIIa while significant ischemic ECG changes were evident in association of aggravation of dysesthesia in the type BIIb patients. 2. Concomitant "cervical spondylosis" with radiographic evidence of abnormalities in cervical vertebrae and associated subjective symptoms accounted for 22.9% of group A and for 71.4% of group B. In no case of type BI was there evidence of such complication whilst all the cases of type BII had this complication. 3. The mode of appearance of pain in patients with cervical spondylosis showing angina like pain resembled to that of angina pectoris in type BII but ECG during attack did not reveal any significant ischemic changes. 4. As for interrelation between findings by selective coronary angiography (26 cases of angina pectoris) and complication of "cervical spondylosis", the complication of "cervical spondylosis" was higher in incidence in the group of cases with low-grade coronary arterial changes (degree of occlusion less than 50%) than in the group with greater arterial changes (degree of occlusion over 50%). The findings described suggest the possibility that the mode of manifestation of anginal attack may be modified by the concomitant presence of "cervical spondylosis".


Thrombus formation around an endocardial pacemaker electrode: a scanning electron microscopic study and its relationship to abnormal threshold elevation. The author experienced an abnormal increase in stimulation threshold in one patient to whom a permanent endocardial electrode was implanted according to the temporary-permanent technique, but could safely exchange the electrode at the fifth postoperative day before exit block would have occurred. Around the extracted electrode there was a thrombus which was considered to be the main causative factor of abnormally increased stimulation threshold. The morphology of the surface of the thrombus and surface of the electrode was examined using a scanning electron microscope, and the author found that the membranous thrombus which surrounded the neck of the lead had a very fine network of configuration with multilayers and that, though macroscopically the electrode tip was clean and the metal was brilliant, there were scattered white masses strongly adhered to the surface of it. Thus in a case of rapid increase of stimulation threshold within a few days after implantation, it is advisable to withdraw the catheter and to implant another new one.


Mean systemic pressure and mean pulmonary pressure: their effects on the total artificial heart. Mean systemic pressure (MSP) and mean pulmonary pressure (MPF), which are mean driving pressures for venous return in the natural heart, were studied in 11 calves in which the natural heart had been replaced with a total artificial heart (TAH). They were measured simply by stopping the artificial heart pumping. Although blood translocation from the arterial to the venous side was not performed, the eventual right and left atrial pressures reached six to eight seconds after stopping the TAH would represent MSP and MPP with reasonable accuracy. The MSP varied from nine to 3k mmHg (20+/-6 mmHg), whereas the MPP varied from nine to 39 mmHg (22+/-7 mmHg). The MSP varied in close relation to the right atrial pressure prior to cessation of the TAH (r=0.9124). Increases in RAP and MSP were mainly attributed to an increase in circulating blood volume. In the performance of the TAH, MSP (or MPP), proper diastolic duration and vacuum application during diastole was of prime importance in determining the end-diastolic ventricular volume.