Suppression of tumor growth in strain 2 guinea pigs by xenogeneic antitumor antibody. Line 10 hepatoma cells were incubated with antiserum specific against line 10 cells (RaL10) and then tested for growth in syngeneic Wright strain 2 guinea pigs. Palpable tumors appeared in only 11 of 23 animals inoculated id with 10(5) RaL10-treated tumor cells, compared with an incidence of 21 of 23 for nonimmune rabbit serum (NRS)-treated cells and 23 of 23 for cells treated with syngeneic guinea pig serum. Animals inoculated with RaL10-treated tumor cells did not develop systemic tumor immunity. The long-term survival of guinea pigs receiving RaL10-treated tumor cells iv was 6 of 12 with a dose of 10(5) cells and 8 of 11 with 10(4) cells. None of the animals receiving 10(4) or 10(5) control tumor cells treated with NRS survived. RaL10 antiserum was not toxic to line 10 tumor cells in vitro, but mediated tumor-specific cytolytic reactions in the presence of fresh guinea pig serum or on the addition of peritoneal exudate cells from nonimmunized syngeneic guinea pigs.


Characteristics of JMV Marek's disease tumor: a nonproductively infected transplantable cell lacking in rescuable Virus. Cells of the JMV Marek's disease (MD) tumor, originally produced by rapid serial passage of MD lymphoma cells in chickens, were characterized to determine whether they were of host or donor origin and to ascertain certain virus-host cell interrelationships. Differences noted in blood group B surface alloantigens between tumor cells and host lymphocytes indicated a probable nonhost origin (i.e., transplantability) of the tumor. JMV spleen tumors contained predominantly large lymphoblasts bearing MD tumor-associated surface antigen. DNA from JMV tumor cell suspensions hybridized significantly with MD virus cRNA, which indicated that JMV cells contained at least a portion of the MD virus genome. No MD virus was rescued from JMV tumors by techniques suitable for rescue of virus from MD lymphomas. The JMV tumor cells were also devoid of MD virus-specific antigens. These properties differed markedly from those of MD lymphoma cells and make the JMV tumor cell a unique, potentially valuable, tool for further study of oncogenic herpesvirus infection and tumor immunity in the chicken.


Biologic characteristics of some mouse mammary tumor viruses. The mammary tumor virus (MuMTV) in the milks of 7 mouse strains and substrains was titrated for infectivity in 4 strains. The data indicated that: 1) Each strain shed a different MuMTV and some genetic strains carried two MuMTV's, each discernible by its mouse strain preference in infectivity tests. 2) Less than 5% of RIIIf and about 10% of Af mice shed detectable MuMTV antigen in their milks after the third parturition. After the sixth parturition, 33% of RIIIf and 50% of Af, and after the ninth parturition, 60% of RIIIf and 90% of Af mice shed viral antigen in their milks. The MuMTV's in milks of high-parity mothers were most infectious in mouse strains different from those most susceptible to MuMTV in RIII and A milks of low-parity mothers. Therefore, RIII and A mice each harbored two viruses, one that was removed by foster-nursing and the other that was not. 3) The susceptibility incidence of RIIIfC57BL mice to RIII virus changed gradually from about 10% in 1970 to about 70% in 1975. Susceptibility of C57BL mice to RIII virus did not change appreciably over this period, and the natural tumor incidence in RIIIfC57BL remained unchanged (about 10%). In addition to their susceptibility to RIII virus, C57BL mice were also susceptible to GR virus; they were relatively resistant to other strains tested. They were especially resistant to RIIIf virus, to which Af and BALB/c mice were very susceptible. 4) Approximately 90% of C3HfC57BL and C3HfBALB/c mice shed antigen in their milks after the third parturition, although the tumor incidence was less and occurred later than in C3H mice. No clear-cut differences could be detected in infectivities between low-parity C3H milk and high-parity C3Hf milks tested in several assay strains.


Characterization of feline T-and B-lymphocytes and identification of an experimentally induced T-cell neoplasm in the cat. Membrane markers of feline T- and B-lymphocytes were identified for further investigation of leukemogenesis in the cat. Feline T-cells formed spontaneous erythrocyte rosettes with guinea pig (GPE) and rat erythrocytes (RE). The receptors for GPE and RE were separate entities and expressed independently on lymphoid cell membranes. The RE receptor appeared to be present only on more mature or differentiated T-cells, whereas the GPE receptor reacted with a broader population that included less differentiated T-cells. Feline B-cells bore a complement receptor that was detected by adherence of SE coated with antibody and complement. Malignant lymphoblasts obtained from thoracic fluid of cats with feline leukemia virus (FeLV)-induced thymic lymphosarcomas, as well as FL-74 cells (a FeLV-transformed feline lymphoblastoid cell line) expressed T-cell markers. These results provided definitive evidence for markers of feline T- and B-cells and identified an experimentally induced T-cell lymphosarcoma.


Fine structure of pancreatic adenocarcinoma induced in rats by 7,12-dimethylbenz(a)anthracene. We induced pancreatic adenocarcinomas in Long-Evans rats by placing crystals, 2-3 mg, of 7,12-dimethylbenz[a]anthracene (DMBA) in a 2- to 3-mm incision in the "head" of the pancreas approximately 1 cm from the duodenum. The incisions were closed with one or two silk sutures. The animals were killed 4-10 months after DMBA implantation, and nodules were removed and routinely prepared for light and/or electron microscopic study. Histologic organization varied from normal, through areas of tubule-like structures, to sheets of pleomorphic tumor cells. Electron microscopic study of tumor cells revealed large electron-lucent nuclei that frequently had irregular outlines and prominent nucleoli. The predominant feature of the cytoplasm was abundant rough endoplasmic reticulum. Zymogen granules were rare. Adjacent cells sometimes were jointed by an apical junctional complex to form a lumen into which projected irregular microvilli. A basal lamina sometimes occurred at the bases of the tumor cells. The fine structural similarity of these tumor cells to acinar cells was noted.


Oncornavirus particles in lymphoid cultures from a howler monkey with Herpesvirus saimiri-induced disease. Budding and extracellular oncornavirus particles were observed in cells of lymphoid cultures derived from the spleen, lymph node, and blood of a howler monkey (Alouatta caraya) that developed a malignant lymphoproliferative disease after infection with Herpesvirus saimiri. The various possible sources of origin of these particles are discussed.


Mononuclear cell fraction carrying Herpesvirus saimiri in persistently infected squirrel monkeys. Circulating lymphocytes from squirrel monkeys persistently infected with Herpesvirus saimiri (HVS) were separated into B- and T-lymphocyte fractions by a rosette-enrichment technique. HVS was isolated only from lymphocyte fractions forming rosettes or from unseparated lymphocytes; this indicated that T-lymphocytes were the target cells for HVS in the natural host, squirrel monkeys.


Immunotherapy of cancer with nonliving mycobacteria and cord factor (trehalose-6,6'-dimycolate) in aqueous medium. Heat-killed BCG, cord factor (trehalose-6,6'-dimycolate), or killed BCG plus cord factor in aqueous medium, admixed with tumor cells and injected into the skin of guinea pigs, inhibited the growth of a hepatocellular carcinoma. Intralesional administration of killed BCG or Mycobacterium kansasii coated with cord factor, in the same medium, caused regression of established tumors in 48 and 45% of the treated animals, respectively.


Ascites form of a human cancer serially transplantable in nude mice. Human adenocarcinoma cells injected into the peritoneal cavities of BALB/c nude mice (nu/nu) induced ascites carcinoma. The inoculant was obtained from subcutaneous tumors produced in nude mice by an injection of ascites cells from a patient with carcinomatous peritonitis caused by mucinous adenocarcinoma of the stomach. An ascitic fluid began to accumulate 45 days after inoculation and reached the maximum volume within 120 days. Dispersed stomach cancer cells in the ascites could be serially transplanted in nude mice in an ascites form. The morphology of these cells was similar to that of the original cells in the ascitic fluid of a patient with carcinomatous peritonitis.


Burkitt's lymphoma: its clinical course in relation to immunologic reactivities to Epstein-Barr virus and tumor-related antigens. In 141 patients with African Burkitt's lymphoma, the relationship between Epstein-Barr virus (EBV)-related antibody titers and the clinical course of this disease was presented. Antiviral capsid antigen tests gave positive results in all patients, siblings, and control neighbors; but the geometric mean antibody titers to viral capsid antigen were significantly higher in patients than in siblings or neighbors (P less than 0.001). No control neighbors or siblings had antibodies to restricted (EA-R) or diffuse (EA-D) early antigen. Mean geometric anti-EA-R titers at admssion and at last visit were significantly lower in patients with stage (I and II) than in those with stage (III and IV) disease; this most likely reflected the degree of tumor burden. Patients who relapsed after 1 year of sustained remission had significantly higher anti-EA-R titers than did those who did not. The increase in the probability of relapse was sixfold for those patients with an anti-EA-R titer of greater than 160 after 1 year of sustained remission. Survivors and nonsurvivors differed significantly in the final EA-R and Epstein-Barr virus nuclear antigen (EBNA) titers (P less than 0.05 and P less than 0.001, respectively). Anti-EA-D titers were particularly likely to be positive in patients with multiple relapses. When skin reactivity to an antigen from RAJI cells was compared to EBV-related serologic reactions in the same patient, a significant inverse correlation (P less than 0.001) between skin reactivity and EBNA titers appeared. Pretreatment sera from patients with high EBNA titers did not block skin reactivity to the RAJI antigen.


Long-term effects of neonatal steroid exposure on mammary gland development and tumorigenesis in mice. Newborn female mice of three strains--BALB/cfC3H [mammary tumor virus (MuMTV)-infected], BALB/c, and C57BL (both virus-free)--were given injections of 17beta-estradiol or testosterone, alone or in combination with ovine prolactin, for the first 5 days of life. Half of each group of mice were ovariectomized at 40 days of age, and all mice were killed between 6 and 16 months of age. Mammary glands of BALB/cfC3H mice receiving steroid hormones were better developed than those of mice not receiving steroids. Androgen induced a higher incidence of grossly dilated ducts and secretion-filled alveoli. Mammary nodule and tumor incidences were higher in steroid-treated mice than in controls; androgen resulted in higher incidences than did estrogen. The age of onset of mammary tumors was also earlier after neonatal steroid treatment. In BALB/c mice, neonatal injections of estrogen induced some alveolar development of the mammary gland; neonatal injections of ovine prolactin had a greater effect. The mammary glands of C57BL mice did not show any evidence of stimulation by neonatal hormone treatment, which indicated the probability of strain differences. However, no nodules or tumors occurred in these MuMTV-free strains. Therefore, MuMTV was essential for neoplastic mammary responses to neonatal hormone treatment. Ovariectomy prevented alveolar development and abnormal changes in the mammary glands of all groups, thus indicating that ovary-independent alterations in the mammary gland were not induced by neonatal steroid treatment. We concluded that neonatal steroid exposure resulted in increased mammary tumor risk in mice, but only in the presence of both MuMTV and ovaries.


Induction of lymphoma and associated xenotropic type C virus in C57L mice by whole-body irradiation. Adult C57L mice received sublethal whole-body X-irradiation. Between 3 and 11 months later, 5 of the 7 exposed mice developed histopathologically confirmed malignant lymphomas (lymphocytic type) that primarily involved the thymus. The lymphomas were readily transplantable to other C57L mice of any age, which developed fetal lymphomatous involvement; the tumor cells could also be propagated in tissue culture. A xenotropic murine type C virus (MuX) was isolated from the cultured lymphoma cells after cocultivation with a permissive dog line. MuX activity was demonstrated by electron microscopy, complement fixation, indirect fluorescent antibody, infectivity, and genome rescue.


Lymphocyte mitogen reactivity and enumeration of circulating B- and T-cells during feline leukemia virus infection in the cat. Mitogen-induced blast transformation of peripheral blood lymphocytes and quantitative changes in circulating T- and B-cells were studied serially in cats inoculated with feline leukemia virus (FeLV). Concanavalin A-induced blast transformation sharply declined beginning at 5 weeks post inoculation (Pl) in FeLV-infected cats when compared to age-matched uninfected control cats. Similar but less consistent changes were seen in responses to pokeweed mitogen-induced stimulation. In most infected kittens this defect persisted until they died from thymic lymphosarcoma, 15-24 weeks Pl. An early lymphopenia, due primarily to a decrease in circulating B-cells, occurred in infected cats 5-8 weeks Pl. Following a return of total and B-lymphocytes to control values, infected cats developed increased numbers of T-cells at 16 or more weeks Pl, which correlated with circulating lymphoblastic lymphocytes bearing T-cell markers. These results correlated neoplasia arising in a thymus-derived lymphocyte population with mitogenic hyporeactivity in the preneoplastic period and suggested that FeLV-induced immune alterations may be a necessary antecedent of leukemogenesis in the cat.


Epithelial characteristics of tumor cells in nasophasyngeal carcinoma. Electron microscopy examination of nasopharyngeal carcinoma grafts in nude mice showed that the tumor cells retained their epithelial characteristics, as betrayed by the presence of keratin fibrils and of functional complexes at the cell membrane. Thus the cells positive for Epstein-Barr virus-specific nuclear antigen present in such grafts do represent the progeny of human epithelial tumor cells. C-type virus particles were observed in one tumor graft, indicating that epithelial human cells could pick up a mouse virus.


Membranes of normal hamster lymphocytes and lymphoid cells neoplastically transformed by simian virus 40. I. High-yield purification of plasma membrane fragments. In this first paper of a series comparing the membranes of normal lymphocyte populations from male outbred Syrian hamsters with those of neoplastic transformants (GD 248) induced by simian virus 40, a method is described for the isolation of representative plasma membrane (PM) fragments from both cell types. Multiple criteria were used to monitor the purity and yield of PM material after cell disruption by nitrogen cavitation and after membrane fractionation by a combination of differential centrifugation and isopyknic ultracentrifugation in dextran density gradients. Lactoperoxidase-catalyzed radioiodination before cell disruption was used as an extrinsic surface marker; Na+,K+-activated ATPase, as well as alkaline phosphatase, was used as intrinsic functional PM markers. The distribution of nuclei, mitochondria, lysosomes, and endoplasmic reticulum (ER) during fractionation was monitored by the measurement of DNA, succinate dehydrogenase and monoamine oxidase, beta-glucuronidase and glucose-6-phosphatase, and NADH:lipoamide oxidoreductase, respectively. According to the three PM markers employed, a 15- to 20-fold purification (over homogenate) and a PM yield of about 65% were obtained for both cell categories, with negligible contamination by DNA, mitochondria, lysosomes, and er. The procedure also allowed recovery of 60% of the mitochondria free of other cell elements.


Membranes of normal hamster lymphocytes and lymphoid cells neoplastically transformed by simian virus 40. II. Plasma membrane proteins analyzed by dodecyl sulfate-polyacrylamide gel electrophoresis and two-dimensional immune electrophoresis. The plasma membrane proteins of lymphocyte populations from normal outbred Syrian hamsters were compared with those of a neoplastic transformant line (GD 248) induced by simian virus 40. Both quantitative and qualitative differences were observed. Gradient dodecyl sulfate-polyacrylamide gel electrophoresis revealed 12 major protein components in the membranes of both cell populations. Both membrane categories also contained small amounts of immunoglobulin. Compared with the membranes of the reference cell population, GD 248 membranes showed a 60% decrease of approximately 210,000 daltons of glycoprotein; a 10% reduction of about a 48,000-dalton band and virtually complete loss of a 15,000-dalton component concomitant with a 57% increase in a 52,000-dalton band; fusion to two subcomponents (mol wt approximately 250,000 daltons); and emergence of approximately 120,000 and 30,000 daltons glycoproteins. In addition, the relative mobility of an approximately 95,000-dalton component increased by roughly 0.02 U. Crossed immune electrophoresis in Trition X-100 with heterologous antiserum against GD 248 microsomal membranes revealed both a new component with a high level of electrophoretic mobility and intensification and additional heterogeneity in a strongly antigenic component with a low level of electrophoretic mobility. Crossed-line immune electrophoresis indicated that at least two antigens in the membranes of GD 248 cells lacked the membranes of the reference cell population.


A method for the control of lymphoid leukosis in chickens. Lymphoid leukosis (LL) was eliminated from 3 inbred lines of White Leghorn chickens that were temporarily kept in isolation. The method of control was based on three elements: 1) From an infected flock we selected hens that produced LL virus-free eggs. Pooled extracts from groups of embryos were tested for vertical virus transmission by the nonproducer cell activation test. 2) Only eggs from dams that did not congenitally shed virus to their embryos were used to produce progeny. The offspring were reared in isolation for 2 months (at which time the age-related resistance against tumor formation had developed sufficiently). 3) The birds were subsequently inoculated im with 10(5) mean tissue culture infective doses of LL viruses of subgroups A and B and transferred to a conventional chicken house. The controlled exposure provided immunity against LL virus infection in a highly infect environment. This procedure resulted in birds a) with no boservable LL, and b) producing LL virus-free eggs. To obtain flocks that had no LL and produced virus-free progeny, the procedure had to be repeated for at least two generations due to the intermittent virus excretion that often occurred.


Inefficient humoral immune response of lymphoma-prone wild mice to persistent leukemia virus infection. Adult wild mice (LC) from a natural colony with a high incidence of spontaneous lymphomas had free infectious virus in their seara (average 10(3.5) infectious units/ml) and parenchymal organs (average 10(5.2) infectious units/g tissue). They did not have detectable levels of free virus-specific antibodies that could be demonstrated by virus neutralization or immunofluorescence at higher than a 1:10 dilution. Only 5 of 28 animals had free antibodies detectable by radioimmunoprecipitation assay, and tissues of 4 mice also had nondetectable levels of virus determined by infectivity assay. Formalized vaccine from the indigenous virus did not induce production of virus-neutralizing antibodies or protect against naturally occurring disease. The animals with persistent leukemia virus infection, however, elicited good humoral immune responses to virus-unrelated antigen.


Mammary tumorigenesis in chemical carcinogen-treated mice. VI. Tumor-producing capabilities of mammary dysplasias in BALB/cCrgl mice. Two types of mammary dysplasias occurring in 7,12-dimethylbenz[a]anthracene (DMBA)-treated BALB/cCrgl mice were transplanted into the cleared mammary fat pads of syngeneic mice for an assessment of their growth behavior and tumor potentials. Keratinized nodules, numerous in DMBA-treated, pituitary isograft-bearing BALB/cCrgl mice, produced primarily ductal outgrowth in control mice and very few tumors (7%) 56 weeks after transplantation. Such dysplasias transplanted into mice bearing pituitary isografts exhibited lobuloalveolar development and produced a higher incidence of tumors (32%). Hyperplastic alveolar nodules (HAN), though relatively rare in DMBA-treated BALB/cCrgl mice, produced lobuloalveolar outgrowth in control mice and had a 100% tumor incidence. Four HAN outgrowth lines were developed by serial transplantation of samples of the nodule outgrowths. The tumor potentials of these nodule lines in intact controls and ovariectomized mice was determined over several transplant generations. The tumor potentials of two of the three nodule lines were decreased in the absence of ovarian hormones. However, the growth of 23 mammary tumors derived from these nodule lines and of nine derived from in situ primary tumors was unaffected by the absence of the ovary. These results, along with those published previously, suggest that mammary tumors in chemical carcinogen-treated mice arise from several precursor populations. These preneoplastic populations comprise both alveolar and ductal hyperplasias.


An experimental animal model for the study of human scirrhous carcinoma. Scirrhous-like carcinomas were induced by the inoculation of cloned mouse mammary carcinoma cells into the cleared fat pads of BALB/c mice. The inoculated cells induced palpable tumors in 100% of the animals within 60 days. The epithelial cells in vivo grew in rows and clusters and were attached by short developing desmosomes. Microvilli and organelles were scarce; intracisternal type A particles were observed in all the epithelial cells. Carcinoma cells invaded the dermis and muscle. The stroma was formed by morphologically normal fibroblasts and large masses of collagen. No virus particles were observed budding from the epithelial cell surface or in the stromal fibroblasts. The similarities between the scirrhous-like carcinoma in BLAB/c mice and the juman scirrhous carcinoma suggest this animal system as a model for the study of the human disease.


Sequential hepatic histologic and histochemical changes produced by diethylnitrosamine in the rhesus monkey. Six young adult male rhesus monkeys were given diethylnitrosamine ip for 3-5 years. Liver biospies were done monthly. After 6 months, biopsy specimens showed individual hepatocytes and small foci of hepatocytes that were intensely positive for glycogen. During the second and later years, larger foci of such cells developed. In sections stained with hematoxylin and eosin, the glycogen-containing hepatocytes generally appeared unusually clear. Some hepatocytes, however, had eosinophilic or basophilic cytoplasm. Nuclear enlargement and atypic developed, particularly outside the foci. The hepatocytes within most foci were uniform in their histochemical features: glycogen was elevated, glucose-6-phosphatase was decreased, and ATPase activity was present not only along the bile canalicular surface but also along the enire cell membrane. After 3-5 years, neoplastic nodules and hepatocarcinomas developed in 5 of 6 animals. Two nodules and particularly the heptocarcinomas differed from the foci in one of more histochemical parameters. The findings suggested that the glycogen-containing, histochemically altered cells of the foci in one or more histochemical parameters. The findings suggested that the glycogen-containing, histochemically altered cells of the foci may be the first step in the development of neoplasia; further steps toward malignancy appeared to be frequently associated with additional alterations, such as loss of sinusoidal ATPase and re-formation of glucose-6-phosphatase.


Transport, phosphorylation, and toxicity of a tricyclic nucleoside in cultured Novikoff rat hepatoma cells and other cell lines and relase of its monophosphate by the cells. 1,4,5,6,8-Pentaazaacenaphthylene-3-amino-1,5-dihydro-5-methyl-(5-14C)-1-beta-D-ribofuranysly (NSC-154020), a tricyclic 7-deazapurine nucleoside (TCN), was rapidly incorporated into the acid-soluble pool by cultured Novikoff rat hepatoma cells, mouse L-cells, HeLa cells, and HEp-2 cells, but little incorporation into nucleic acids occurred. More than 90% of the intracellular radioactivity was associated with the monophosphate (MP) of the substrate concentration followed normal Michaelis-Menten kinetics. Comparison of the kinetic constants for the uptake of adenosine and the TCN and of the inhibition of their uptake by each other suggests that both were transported by the same system (Michaelis constant approximately 6-10 muM) and with about the same efficiency. The TCN was also phosphorylated in a cellfree extract containing adenosine kinase activity at about the same rate as was adenosine, but not further phosphorylation of the analogue MP occurred. No significant deamination or degradation of the adenosine analogue to its base and ribose-1-phosphate was observed. TCN inhibited the replication of all four types of cells propagated in suspension culture; however, Novikoff cells were several times more sensitive than were the other three cell types, despite the finding that the TCN-MP, probably the main toxic principle, accumulated to about the same concentration in cells of all four lines. Complete inhibition of replication of Novikoff cells were several times more sensitive than were the other three cell types, despite the finding that the TCN-MP, probably the main toxic principle, accumulated to about the same concentration in cells of all four lines. Complete inhibition of replication of Novikoff cells and cell death occurred at concentration as low as 15 muM TCN. At these concentrations, TCN, within 2 hours of its addition, completely inhibited the incorporation of [14C]formate int0 nucleotides and nucleic acids of Novikoff cells. An inhibition of the denovo synthesis of purine and pyrimidines, however, was not the only toxic effect of the TCN since high concentrations of uridine, adenine, guanine, and hypoxanthine, either alone or combined, failed to prevent the inhibition of cell replication by TCN. Also, between 1 and 3 hours of treatment, 70-80% of the Novikoff cells fragmented into four to eight vesicles per cell. These fragments were impermeable to trypan blue, still exhibited some metabolic activity such as the phosphorylation of AMP and TCN, but failed to replicate when the drug was removed. No similar fragmentation was observed with the other cell lines. Novikoff and L-cells rapidly released TCN-MP into the culture fluid. After 4 hours of incubation, 70-100% of the total radioactivity in the medium was associated with the MP. Only a little TCN-MP was released from HeLa and HEp-2 cells. A TCN-resistant mutant of Novikoff cells failed to phosphorylate the analogue and was deficient in adenosine kinase.


Reduction of rat liver carcinogenicity of 4-nitrosomorpholine by alpha-deuterium substitution. Groups of 30 males Sprague-Dawley rats were given 4-nitrosomorpholine-3,3,5,5-d in their drinking water at concentrations of 0.35 and 0.07 X 10(-3) M for 30 weeks. Two similar groups of rats were simultaneously given unlabeled 4-nitrosomorpholine (NM) at the same malar concentrations; all animals were observed throughout their lives. Those receiving the alpha-deuterium-labeled compound had significantly fewer liver tumors than did the corresponding animals receiving the unlabeled compound. The difference in potency appeared to be at least fivefold, a magnitude consistent with a primary kinetic isotope effect on the carcinogenic action of NM. Thus breakage of a bond linking a hydrogen (deuterium) atom with carbon adjacent to the nitrosamino function may be involved in a rate-limiting step of carcinogenesis by NM.


Carcinogenicity test of two unstaurated derivatives of N-nitrosopiperdine in Sprague-Dawley rats. Two unsaturated derivatives of N-nitrosopiperidine (NP), N-nitroso-1,2,3,6-tetrahydropyridine (NTP) and N-nitrosoguvacoline (NGC), were administered to Sprague-Dawley rats as solutions in drinking water at concentrations equimolar (0.88X10(-3) M) with those used to test NP. NTP gave rise to hepatocellular tumors in contrast to the esophageal or olfactory tumors that were induced by NP. NGS did not induce tumors under these test conditions.


Hepatocarcinogenicity of estragole (1-allyl-4-methoxybenzene) and 1'-hydroxyestragole in the mouse and mutagenicity of 1'-acetoxyestragole in bacteria. Approximately 20% of a dose of estragole, a naturally occurring flavoring agent, was excreted in the urine of outbred male CD -1 mice as a conjuage (presumably the glucuronide) of 1'-hydroxyestragole, Estragole and its 1'-hydroxy metabolite caused significant increases in the incidences of hepatocellular carcinomas in male CD-1 mice that received the compounds by sc injection at 1-22 days of age. Estragole induced hepatocellular carcinomas by 15 months in 23 and 39% of the mice that received total doses of 4.4 and 5.2 mumoles, respectively, and lived to an age of 12 months or more. Of the 12-month survivors given a total dose of 4.4 mumoles of 1'-hydroxyestragole, 70% developed hepatocellular carcinomas; the incidence in mice that received only the vehicle (trioctanoin) was 12%. Multiple tumors ocurred in 5, 28, 64, and 0%, respectively, of the mice in each of these 4 groups. Of the mice given a total dose of 4.4 mumoles of 1'-hydroxysafrole, 59developed hepatocellular carcinomas; 39% of the mice bore multiple liver tumors. As previsously demonstrated for 1'-acetoxysafrole, 1'-acetoxyestragole and 1'-acetoxy-1-allyl-4-methoxynaphthalene reacted nonenzymatically with guanosine and inosine to form adducts. These electrophilic esters were strongly mutagenic for the Salmonella typhimurium missense mutant TA100. 1'-Acetoxyallybenzene had little or no activity in either of these tests. Attempts to demonstrate liver-mediated mutagenicty for 1'-hydroxysafrole and 1'-hydroxyestragole in the bacterial test system were unsuccessful.


Endogenous oncornaviruses in chemically induced transformation. II. Effect of virus production in vivo. C3H/HeJ and AKR/J mice differed in their susceptibility to 3-methylcholantrhene (MCA)-induced sarcomagenesis (86% incidence of sarcomas in C3H by 18 wk; 5% incidence in AKR by 18 wk) and in the production of endogenous murine leukemia virus (MuLV) (AKR produced greater than 10(5) plaque-forming units/ml tail extract in XC test; C3H did not produce detectable virus.) A genetic corss between C3H and AKR mice was examined to determine the relationship of virus production to oncogenesis by MCA. Mice of the (C3H X AKR)F X C3H backcross were typed for the production of infectious MuLV by tail biospy and then inoculated with MCA. Of the backcross mice, 81% produced high titers of ecotropic MuLV; the remaining 19% did not contain detectable infectious MuLV. The virus-producing and non-virus-producing backcross mice were equally sensitive and highly susceptible to MCA-induced sarcomagenesis. Tumors of all virus-positive mice contained infectious MuLV. Some tumors (54%) of virus-negative mice also contained infectious MuLV; this indicated the induction of endogenous MuLV in the tumors of these mice. We concluded that the overt production of MuLV in mice of this backcross did not function in the sensitivity of the mice to sarcoma induction by MCA. Furthermore, the presence of virus in some chemically induced tumors was due to an induction pehnomenon independent of the primary oncogenic event.


Promotion of tumor growth in vivo by antimacrophage agents. Various attempts were made to assess the role of the mononuclear phagocyte system in tumor resistance of rats in vivo. The growth of sc inoculated, weakly immunogenic, carcinogen-induced, syngeneic tumor cells was modestly reduced by ip injection and, more impressively, by local injection of peptone-induced, activated, nonimmune macrophages. A single iv injection of silica particles or carrageenan on the day of sc tumor cell inoculation greatly enhanced tumor growth. When these agents had been given a few days before or after tumor cell inoculation, the tumor-promoting efficiency was distintly diminished or even cancelled. The enhancing effects of silica and carrageenan on tumor growth were nulified by the macrophage-stabilizing agent, poly-2-vinylpyridine N-oxide, To assess the in vivo consequences of silica administration, various cellular, biochemical, and functional macrophage parameters were determined at different intervals. Results indicated the complexity of events elicited after the mononuclear phagoycte system was damaged, which made the interpretation of such results difficult.


Combination chemotherapy of L1210 leukemia with platinum compounds and cyclophosphamide plus other selected antineoplastic agents. Six antitumor platinum compounds were used in combination with cyclophosphamide plut 1 of 7 other antitumor drugs for treatment of L1210 leukemia in B6D2F [C57BL/6 X DMA/2) F] mice. Data obtained from each three-agent regimen were compared with those obtained after administration of each compound alone and each appropriate two-agent combination. No cure (greater than 60-day survival) was obtained with any compound used alone. Combination of cyclophosphamide with a platinum compound (Pt+CY) yielded a collective cure rate of 193/420, and the addition of a third cure rate to 290/420 (P less than 0.001). Certain regimens produced 100% cure rates. The most effective drugs when used in combination with PT+CY were cytosine arabinoside, 5-fluorouracil, hydroxyurea, and Yoshi-864. Adriamycin, methotrexate, and vincristine were less effective at the doses used. Toxicity, as evidenced by maximum weight loss, was slightly greater with the three-agent combinations than with the Pt+CY regimens.


Effect of adrenergic agonists on the cyclic AMP level in the liver and heart from normal and hypothyroid rats. In order to clarify the mechanism of the different sensitivity of the adrenoceptors between normal and hypothyroid rats, cyclic AMP levels in the liver and heart were measured after the administration of phenylephrine, isoproterenol, epinephrine and methoxamine. Cyclic AMP increased in all cases, but the extent of its increment in the heart was much less than that in the liver. Phentolamine and propranolol showed only a partial inhibition of cyclic AMP elevation by the agonists in the liver from normal and hypothyroid rat. On the other hand, propranolol blocked completely the effect of the agonists on the heart from both groups. It was also observed that cyclic AMP increased in adrenalectomized rats after the injection of the adrenergic agonists. The basal activity of protein kinase in hypothyroid status was slightly lower than that in the normal, but this enzyme was stimulated in the presence of cyclic AMP in vitro. These results suggest that the function of beta-adrenoceptor remained normal even in hypothyroidism and responded well to isoproterenol and epinephrine. It is also indicated that the increased sensitivity of alpha-adrenoceptor to phenylephrine in the hy pothyroid atria previously observed is probably in part independent of the mechanism mediated by cyclic AMP.


[Differential diagnostic problems resulting from massive doses of vitamin D and suspected pituitary dwarfism in the father of a child with vitamin-D-resistant rachitis (author's transl)]. A 1.8-year-old female child with retarded statomotor development and growth was reported. Radiologically and chemically, a rachitis was found which clearly improved following administration of 600,000 U. vitamin D3. Examination of the father who was thought to be suffering from pituitary dwarfism revealed hypophosphatemia and radiologic signs of osteomalacia. The diagnosis of a hypophosphatemic vitamin-D-resistant rachitis in this child could only be established with certainty during the course of the following months.


[Clinical and diagnostical aspects of acute cytomegalovirus infections in previously healthy adults (author's transl)]. The clinical symptoms of floride Cytomegalovirus infections of 18 previously healthy adults are reported. Fever was generally observed. In most cases myocarditis or hepatitis could be identified. Two of our patients died of on severe myocarditis. In 3 patients Guillain-Barré-syndrome was observed. Only in few of our cases the clinical picture was characterized by mononucleosis with systemic swelling of lymphonodes and thrombocytopenia or by pneumonia. Other symptoms were found quite seldom. The diagnosis of cytomegalovirus infection could be ascertained by: 1. detection of cytomegalovirus IgM antibody titers of greater 1:64, 2. a four fold increase in the titer of complement fixing antibodies, 3. Virus isolation from urine.


Properties of calf endothelial cells in culture. Calf aorta endothelial cells, obtained by collagenase treatment of vessels from freshly killed animals, were cultured in medium 199 supplemented with fetal bovine serum, amino acids, and vitamins. The material released from the vessel wall after collagenase treatment consists of clumps of cells which quickly attach to the dish and spread to form confluent islands of cells. These islands of cells coalesce to form a confluent monolayer in 6 to 8 days. Cultures labeled with [3H]thymidine show an increase in labeling commencing 3 days after initial culture. Cells within monolayers do not overgrow one another and retain their epithelioid appearance after subculture. Identification of endothelial cells was based on culture morphology and by the presence of factor VIII antigen as localized by indirect immunofluorescence microscopy. These data indicate that large numbers of endothelial cells (5 to 7 x 10(6) cells per aorta) can be obtained and maintained in culture.


Cyclic nucleotides, thioldisulfide status of proteins, and cellular control processes. It is shown that cyclic nucleotides can have a variety of effects on cell division, cell shape, cell adhesion, and cell movement, depending on the cells selected and the conditions under which they are used. For example, while CHO cells elongate under the influence of exogenous dibutyryl CAMP, Y-1 adrenal tumor cells round up and polyoma-transformed 3T3 cells show no change in shape. The totality of experience with cyclic nucleotides suggests that where they have been used by cells as control elements involving the four processes listed above, they are superimposed on basic cellular processes that progress in their absence--that is, they must be acting indirectly. In attempting to understand the inhibitory action of methyl xanthines on egg development, we were forced to abandon the idea that they acted through cyclic nucleotides. We found that methyl xanthines inhibited the activation of glutathione reductase and that glutathione oxidizing agents act as mitotic inhibitors. Further, we found that tubulin polymerizability, NAD-kinase activity, and a mitotic apparatus associated Ca+2-ATP-ase were all inhibited by oxidation of some of their sulfhydryls and were activated by reduction of the resulting disulfides. These results are discussed in terms of reported cycles and activations of glutathione reductase (GR) in cells and reports that mixed disulfides of glutathione and proteins can act as substrates for GR. Using the fact that a CAMP-dependent protein kinase has been reported to be activated by glutathione, we have suggested potential sites where sulfhydryl control processes and cyclic nucleotide control processes and cyclic nucleotide control processes may interact in certain restricted cases.


Distribution pattern of radioactive labelled lipiodol-UF following intralymphatic application for therapy. Endolymphatic radiotherapy with 4 mCi32P tri-n-octylphosphate and 1 mCi 131 I triolein LIPIODOL UF has been performed in 75 patients suffering from malignant melanoma of the lower extremity. On the average, 13.3% of the radioactive substance remains in the syringes and connecting tubes. In most patients the radioactive material available for therapeutic irradiation is further reduced due to contamination of operation sheets and swabs (mean: 15.3%). There is, however, still sufficient radioactivity remaining for effective internal irradiation of the lymph nodes. The average radiation dose absorbed by the lymphatic tissue is 90.998 rad. The method is limited by the hazard of radiation damage to the lungs. Almost 80% of these patients had detectable concentrations of radioactivity in the lung fields. The average radiation dose was found to be 299 rad. So far radiation induced fibrosis has bot been observed in this series.


Inhibition of neuromuscular transmission by prostaglandin E1 in the circular muscle of the guinea-pig vas deferens. Pressure increases elicited by contractions of the circular muscle of the isolated guinea-pig vas deferens in response to nerve stimulation were recorded. In contrast to longitudinal muscle which contracted in response to 1--50 pulses, circular muscle responded only to longer trains of pulses (10--500) at a frequency of 10 Hz. Atropine (1.4 muM) caused a slight depression of responses to 100 shocks. Phentolamine at a concentration of 2.6 muM failed to inhibit the response to stimulation, but a higher concentration (53 muM) caused a definite blockade. Guanethidine (25 muM) strongly reduced the responses. With a stimulus train of 100 pulses no inhibition by prostaglandin E1 (PGE1) (0.028 muM) could be demonstrated; however, at a lower number of shocks (20--50) a clearcut depression was observed. The lower the number of pulses the more marked was the depression. The observation that PGE1 failed to block the contractions evoked by noradrenaline (59 muM) suggests a presynaptic inhibitory action of the prostaglandin. It is suggested that noradrenaline is the transmitter in both muscle coats of the guinea-pig vas deferens and that the neuroeffector junctions are sensitive to the effect of PGE1.


The jugular bulb: its anatomic and clinical considerations in contemporary otology. Advances in neurotologic diagnosis and surgery has pointed to the need for a complete reassessment of the relationship and significance of the jugular bulb and sigmoid sinus is discussed and selected temporal bone sections presented to illustrate the varying locations of the jugualr bulb. Considerations in differential diagnosis are presented and examples of surgical procedures are discussed wherein the jugular bulb was encountered and the methods of management are detailed in these instances.


Lesions of the vagus nerve: diagnosis, treatment and rehabilitation. Lesions of the vagus nervous system have devastating effects on the vital functions of coughing, swallowing and phonation. A clear understanding of the vagal correlative anatomy and physiology leads to a practical organization of upper respiratory and digestive tract lesions. A reasoned and thoughtful treatment plan evolves that leads the patient to comprehensive treatment and rehabilitation.


[Electron transport chain of the Candida mycoderma mutant lacking cytochromes b and a+a3]. Two electrontransport chains--"oxygen" and "peroxidase"--were found to operate in the mutant of Candida mycoderma lacking cytochromes b and a+a3; the terminal acceptors of electrons are oxygen and hydrogen peroxide, respectively. The "oxygen" chain lacks cytochromes but contains pyridine nucleotides and flavoproteins; it is inhibited by rotenone and benzhydroxamic acid. The "peroxidase" chain consists of pyridine nucleotides, cytochrome c, and cytochrome c peroxidase; it is inhibited by cyanide.


[Characteristics of the respiratory chain and the oxidative phosphorylation system of mitochondria in the flavinogenic Eremothecium ashbyii strain]. Tightly coupled mitochondria were isolated from cells of the flavinogenic strain of Eremothecium ashbyii collected during the logarithmic and stationary growth phases. The composition of the respiratory chain and characteristics of the energy coupling system are described. The mitochondria show a wide spectrum of metabolic activity and oxidize Krebs cycle compenents and exogenous NADH. The terminal segment of the respiratory chain is represented by a typical cytochrome system. The mitochondria of the ascomycete collected during the logarithmic growth phase are characterized by a relatively high content of cytochromes b and c, a high rate of oxidation of NAD-dependent substrates, the presence of lower homologues of ubiquinone, UQ6 and UQ7, and extremely high sensitivity of respiration to the action of antimycin A, low content of a component sensitive to rotenone, contrasting with the operation of all three sites of phosphorylation. Transition to the stationary growth phase is accompanied with a decrease in the rate of oxidation of all substrates studied and a declined effectiveness of oxidative phosphorylation. The data obtained are discussed in relation to the ability of the cells for "overproduction" of flavins.


[Phospholipase C of fungi and staphylococci]. The activity of phospholipase was studied in the cultural broth and cell extract of 112 strains of fungi and yeasts. The endoenzyme was detected in 19 strains of mycelial fungi, the exoenzyme was found in Mucor hiemalis 50 and Aspergillus niger 117. Phospholipase C of M. hiemalis was purified and compared to phospholipase of staphylococci. The values of Km are 8.9 and 1.07 mM, respectively, for the fungal and staphylococcal enzymes.


[Effect of calcium hypochlorite on Bacillus anthracoides spores]. The sublethal dose of calcium hypochlorite (CH) of 0.2--0.3 mg/ml active chlorine did not cause, after 5 min, morphological changes in the spores of Bacillus anthracoides which could be detected by phase contrast microscopy, or a decrease in the content of dipicolinic acid (DPA) in the spores. Further cultivation of the spores treated with the sublethal dose of CH om MPA resulted in a delay of changes which were typical of normal germination process (swelling, loss of light refraction, decrease in DPA content). The action of the lethal dose of CH (0.2--0.3 mg/ml active chlorine during 1.5 hr or 5.6 mg/ml active chlorine during 1 hr) causes a decrease in light refraction, changes in the dimensions of spores, and a decrease in the content of DPA in the spores by a factor of 4--5. A sharp decrease in the content of DPA in the spores may characterize not only their germination but also their death caused by lethal doses of the chlorine containing disinfectant.


[Paul-Bunnell negative glandular fever (author's transl)]. Cytomegalovirus infections may occur either as congenital and neonatal diseases respectively or postnatally in adults in connection with conditions of antibody or immunologic deficiencies. Reports of postnatally acquired cytomegalovirus infections in apparently healthy adults are scarce. A case of an acquired cytomegalo-virus infection in a 23-year old male patient who never was ill before is reported. Clinical and laboratory features showed a typical course of a Paul-Bunnell negative glandular fever. Diagnosis could be established by detection of cytomegalovirus IgM serum levels by means of immunofluorescence technique.


Genetic, endocrine, and viral aspects of AKR leukemogenesis. In the AKR strain of mice a high incidence of spontaneous lymphoid leukemia develops before 12 months of age. Genetic, viral, and endocrine factors interact during development to produce the syndrome. Many of the deficiencies of AKR mice are associated with gene action in chromosome 17, at or near the major histocompatibility locus. It is suggested that low steroid levels and high thyroxin levels during an early period of development are an essential part of the syndrome. Specifically, induction of a polymerase enzyme and regulation of the extent of its action by hormones are postulated to favor appearance of C-type particles and accumulation of a population of undifferentiated lymphoid stem cells. Immunodeficiency and leukemic transformation result.


Provocation poliomyelitis and entry of poliovirus to the CNS. It is proposed that entry to the CNS by poliovirus occurs from multiple sites which are seeded during viremia. At each site, an inflammatory reaction allows entry to peripheral nerves. Differences in incubation times are related to the length of nerve to be followed. Virus reaches the CNS sequentially but does not in natural poliomyelitis spread within the CNS. Provoking agents are identified as immunogenic and their action explained. Details of paralysis among Cutter vaccinees are explained by this model, as well as the effects of multiple inoculation in monkeys. Clinical symptoms in the preparalytic stage of poliomyelitis are consistent with this model.


Inhibition of EB virus transformation of non-adherent human lymphocytes by co-cultivation with adult fibroblasts. Transformation of a special population of non-adherent human lymphocytes by EB virus (EBV) is reversibly inhibited by co-cultivation on adult human fibroblasts. Neither fluid from adult fibroblast cultures nor extracts of fibroblasts inhibited such transformation, and the growth of already transformed lymphocytes was not inhibited on adult fibroblasts. The EBV-associated nuclear antigen (EBNA) was detectable in inhibited cultures, with a maximum level of about 15% at 14 days after which it decreased gradually. Reversal of inhibition at 28 days, by either addition of phytohaemagglutinin or by removal of the lymphocytes from the adult fibroblasts, resulted in a prompt increase in the percentage of EBNA-positive cells and typical lymphoblastoid outgrowth. Transformation of EBV-infected non-adherent lymphocytes could be inhibited by the addition of adult fibroblasts up to 2-4 days after infection. The results indicate that, in the EBV-infection of non-adherent lymphocytes on adult fibroblasts, a block resulting in inhibition of transformation occurs between the production of EBNA and the onset of autonomous proliferation of the infected lymphocytes.


XC cell fusion by murine leukemia viruses: fusion from without. Concentrated murine leukemia virus (MuLV) or MuLV producing cells induce XC cell fusion within an hour leading to syncytia formation. While MuLV inactivated by UV irradiation, beta-propiolactone or hydroxylamine treatment still caused cell fusion, Bromelin- or trypsin treated MuLV was no longer able to fuse XC cells. Though sonicated MuLV induced no XC cell fusion, it interfered with cell fusion as caused by untreated MuLV. XC cells infected by diluted MuLV of a titer lower than 1 X 10(5) PFU/ml formed no syncytia although they produced MuLV. The cell fusion mechanism is discussed.


Neutralizing serum IgM antibodies in infections with Herpes simplex virus hominis. Forty-two patients with herpes simplex virus infections were tested for neutralizing serum IgM antibodies. A technique in which serum antibody fluorescein staining was combined with sucrose gradient centrifugation facilitated the isolation of the serum IgM fraction for the use in neutralization tests. In nearly all cases with primary infection, especially those presenting heavy clinical signs (encephalitis/meningitis) the IgM tests were positive. In one case we could detect the IgM antibodies for 11 weeks after the onset of the illness, in another case in cerebrospinal fluid samples for 6 weeks. In localized herpes infections, which were mostly due to reactivations, serum IgM antibodies could only rarely be demonstrated. Among the serologic tests used in this study (NT, CFT, IFT, ACIFT), only the CFT beside the NT (with certain reservations) can be applied for subtyping HSV serum antibodies.


Recurrent genital Herpes simplex virus (HSV) infection of guinea pigs. Intravaginal infection of guinea pigs with HSV type 2 induced lesions resembling genital herpes in humans. Chronic latent infection with spontaneously recurring genital lesions developed in animals surviving the primary disease. Clinical observations and virological studies during the acute and chronic infection are reported.


Serum modulation of in vitro cellular immune reactivity to Epstein-Barr virus-associated antigen. The reactivity of peripheral blood leukocytes to Epstein-Barr virus-associated antigen was assessed employing two in vitro tests, leukocyte adherence inhibition (LAI) and leukocyte migration inhibition (LMI). In both tests, autologous serum from some normal subjects was shown to confer strong reactivity on weak or non-responder cells. Reactivity to purified protein derivative of tuberculin (PPD) assessed in parallel was shown to be unaffected by autologous serum. Employing the LAI system, variations in the effects of autologous serum samples collected on consecutive days was shown to be dependent on the reactivity of the leukocytes themselves, rather than changes in the activity or concentration of factors present in the serum.


The location of the feedback-specific region with the pyrimidine-3 locus of Neurospora crassa. The purimidine-3 locus of Neurospora crassa specifies two enzyme activities, pyrimidine-specific carbamyl phosphate synthetase (CPSpyr) and aspartate transcarbamylase (ATC). ATC is translationally distal. CPSpyr, but not ATC, is subject to feedback inhibition by uridine triphosphate (UTP). To investigate the location of the feedback-specific region within the locus, inhibition of a number of pyr-3 alleles by UTP was investigated. All CPS+ ATC- polar alleles, revertants of CPS- ATC- polar alleles, and 5-fluorouracil-resistant mutants had normal UTP response. The location of the feedback-specific region is in or close to the CPS-specific region.


An ESR study of the postsynatpic membrane acetylcholinesterase of Torpedo marmorata electric organ. The effect of the cholinergic activator, phenyltrimethylammonium, on the ESR spectra of spin-labeled membrane bound acetylcholinesterase was studied; a reduction of maximal hyperfine splitting of the anisotropic ESR spectrum by 2 G was observed. The influence of phenyltrimethylammonium was prevented by the two cholinergic blocking agents d-tubocurarine and alpha-cobratoxine. The present results indicate that the conformation change of the esteratic site of membrane acetylcholinesterase is triggered by the binding of phenyltrimethylammonium to the cholinoreceptor site.


Hormonal control of fat cells adenylate cyclase. Fat cells were preincubated for 2 h in the presence and absence of growth hormone (GH) and Dexamethasone (Dex) before the addition of increasing concentrations of either epinephrine, theophylline or glucagon and final incubation of the cells for an additional 5 minutes. GH and Dex increased by 85%, 28% and 72%, respectively, the cAMP levels reached in the sole presence of 10(-5)M epinephrine, 10(-2)M theophylline or 5 X 10(-5)M glucagon. An adenylate cyclase particulate preparation shows that epinephrine decreases Km from 2mM to 0.6 MM and increases Vmax and the strength of interaction value (n) from 0.91 to 1.75.


[Therapy of myasthenia (author's transl)]. The types of symptoms of myasthenia (Ossermann), specific problems of myasthenia therapy and the fundamental principles of treatment with cholinesterase inhibitors are discussed. In addition to the improved techniques in the observation room, the consistent performance of an immunosuppressive therapy usually leads to improvement of the prognosis of severe myasthenia. The principles of cortisone and Imurek therapy are commented on and the indication for thymectomy are discussed.


Methyl methanesulfonate mutagenesis of synchronized Chlamydomonas. Methyl methanesulfonate (MMS) mutagenesis of Chlamydomonas reinhardtii at different stages of the synchronous cell-cycle revealed the following results. (1) Induction of phenotypically distinct Mendelian (nuclear), str-50 and non-Mendelian (chloroplast) str-500P, streptomycin resistant mutants was relatively high during the first portion of the cell-cycle when chloroplast DNA replication is known to occur. (2) A second and more pronounced interval of enhanced Mendelian, str-50 mutant induction was observed near the middle of the cell-cycle when the initial stages of nuclear DNA replication occur. Induction of non-Mendelian, str-500P mutants was inconsistent during this period. (3) The incidence of mutants from a second phenotypically distinct class of non-Mendelian streptomycin-resistant mutants (str-500D) was not increased over control levels at any stage of the cell-cycle examined. It is concluded that MMS, like N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), may not be the most suitable general mutagen for this alga because its enhanced mutagenesis of cells in the nuclear S phase could result in multiple closely linked mutations.


Studies of factors influencing the mutagenicity of EMS in mice. Three different routes of administration of ethyl methanesulphonate (EMS) (i.p., oral, i.t.) were compared for their relative efficiencies in the induction of dominant lethal effects. Included in the comparisons between oral and i.p. injections, was a preliminary study into the existence of strain differences in sensitivity to EMS between C3D2 F1 hybrid mice and strain DBA/2J mice. No route of administration dependent  effects were found between oral and i.p. injections regardless of the test animal used. I.t. injections of EMS did not induce dominant lethal effects. One treatment related strain difference was observed.


Quantitative evaluation of Epstein-Barr-virus-infected mononuclear peripheral blood leukocytes in infectious mononucleosis. We devised a quantitative assay for Epstein-Barr-virus-infected mononuclear leukocytes (virocytes) to determine their prevalence in the blood of patients with acute-phase and convalescent-phase infectious mononucleosis and in healthy Epstein-Barr-virus-seropositive controls. Mononuclear peripheral blood leukocyte suspensions were tested for virus-determined cytoproliferative activity by cocultivation with human cord-cell indicator cultures. The highest levels of virocytes among circulating mononuclear leukocytes were found in the early acute phase of infectious mononucleosis (up to 0.05 per cent). Virocytemia decreased to levels comparable with those of healthy controls (less than 0.00001 per cent) by the third month after onset of infectious mononucleosis. These findings provide a quantitative profile of the course of the infection at cellular level and support existing evidence of the efficiency of immune control mechanisms in limiting Epstein-Barr-virus infection during the course of infectious mononucleosis.


Transformation by murine and feline sarcoma viruses specifically blocks binding of epidermal growth factor to cells. Normal cells in culture have membrane receptors for epidermal growth factor (EGF); EGF stimulates cells to divide by binding to these receptors. Cells transformed by murine and feline sarcoma viruses rapidly lose the ability to bind EGF, whereas cells transformed by the DNA tumour viruses, polyoma and SV40, or infected with non-transforming RNA tumour viruses have normal levels of functional EGF receptors. The results suggest that a product of the sarcoma virus genome specifically changes cell EGF receptors; the sarcoma gene product may, then, be functionally related to EGF.


Alterations of dopaminergic neurotransmission after chronic morphine treatment: pre- and postjunctional studies in striatal tissue. Repeated morphine administration reversed the acute effects of morphine in rats, e.g. catalepsy and akinesia, and induced symptoms suggesting an activation of dopaminergic mechanisms. In morphine-withdrawn rats, the potency or intrinsic activity of dopamine in stimulating the synthesis of cyclic AMP in striatal homogenates was not significantly altered. However, the K+-induced release of 14C-dopamine from striatal slices of morphine-withdrawn rats was significantly increased, compared with that from striatal slices of saline-treated controls. The results suggest that chronic morphine administration to rats increases the dopaminergic neurotransmission in brain by a pre-synaptic (prejunctional) mechanism, probably reflecting some kind of adaptation to the acute morphine action, which decreases the dopaminergic neurotransmission. The nigro-straital dopaminergic system, therefore, seems to be a good model to study acute morphine actions and mechanisms of morphine dependence at the cellular level.


Influence of papaverine derivatives on phosphodiesterase activity, cyclic 3',5'-AMP levels and relaxing effect on rabbit ileum. The correlations between the relaxing effect of papaverine derivatives, inhibition of low Km-phosphodiesterase (cAMP-PDE = EC 3.1.4.17) activity and cyclic 3',5'-AMP (cAMP) levels in isolated rabbit ileum were investigated. There was a strong correlation between the relaxing effect, inhibition of PDE activity and cAMP content for eupaverine, ethylpapaverine and papaverine. Eupaverine was the most effective relaxing agent (I50 = 7.5 muM) and the most potent inhibitor of PDE activity (Ki = 0.6 muM), followed by ethylpapaverine (I50 = 10 muM;Ki 0.8 muM) and papaverine (I50 = 20 muM;Ki = 2 muM). In contrast, there was a strong relaxing effect (I50 = 6 muM) but only slight inhibition of PDE activity (Ki = 350 muM) by tetrahydropapaveroline (THP). The adenylate cyclase stimulating effect of THP which was shown by others is most likely the reason for comparatively higher cAMP levels, which were found to be elevated about seven times over basal levels of 0.35 nmoles/g wet weight, and effective relaxation. Relaxation could be induced by exogenously added cAMP (I50 = 45 muM) and dibutyryl-cAMP (I50 = 450 muM). Our results support the assumption that smooth muscle relaxation in rabbit ileum is mediated by cAMP. Some of these observations have been published in abstract form (Schulz and Berndt, 1972).


Dopaminergic stimulants and cyclic nucleotides in mouse brain. Effects of dopaminergic antagonists, cholinolytics, and GABA agonists. The effects of dopaminergic stimulants on the cyclic GMP content in the medial forebrain and the cerebellum were studied in mice pretreated with dopaminergic antagonists, cholinolytics and agents enhancing GABAergic transmission. Low doses of butyrophenones (haloperidol and spiroperidol) inhibited the rise in cyclic GMP levels and the stereotyped behaviour induced by amphetamine, but were without effect on the same biochemical and behavioural changes elicited by apomorphine. Higher doses effectively blocked the rise in cyclic GMP levels and the stereotyped behaviour elicited by both drugs. These findings suggest that low doses of the dopaminergic antagonists may predominantly act by interfering with the release of dopamine from presynaptic stores, while high doses may act by blockade of the postsynaptic dopaminergic receptor. The rise in cerebellar cyclic GMP levels elicited by dopaminergic stimulants appears not to involve cholinergic transmission, since atropine did not block the effects of the dopaminergic stimulants. Enhancement of GABAergic transmission by diazepam or aminooxyacetic acid antagonized the rise in cerebellar cyclic GMP content induced by the dopaminergic stimulants, but was without effect on the cyclic GMP content in the medial forebrain. Cyclic AMP levels were not affected by any of the drugs in both parts of the brain.


Effects of methacholine, histamine and atropine on pulmonary guanosine-3', 5'-monophosphate levels in hypersensitive mice. Pulmonary levels of cGMP and cAMP in mice sensitized to methacholine and histamine with B. pertussis were examined to determine whether sensitization could be the result of an alteration in the metabolism of these cyclic nucleotides. The results presented show that in sensitized mice, methacholine raised cGMP to levels that were about double those produced without sensitization. In analogous experiments, histamine raised cGMP by approximately 100% in sensitized mice without producing significant increases in nonsensitized groups. Atropine completely blocked the cGMP rises produced by methacholine but did not eliminate those produced by histamine, thus indicating that cholinergic, but not the histaminergic elevation of cGMP involves activation of muscarinic receptors. The influence of pertussis on cAMP appeared to be opposite in direction from cGMP, i.e., a small but significant drop in cAMP levels was found following methacholine administration to sensitized, but not to nonsensitized mice. It was concluded that pertussis sensitization increases the responsiveness of the pulmonary guanylate cyclase-cGMP system to methacholine and histamine, and that the altered patterns of cGMP accumulation may contribute to the biochemical mechanism of sensitization.


Comparative studies on isolated rat hepatocytes and AS-30D hepatoma cells with leucocidin from Pseudomonas aeruginosa. Leucocidin, one of the cytotoxic principles of Pseudomonas aeruginosa induces potassium loss and swelling in isolated hepatocytes and in AS-30D ascites hepatoma cells in a dose and time dependent manner. Hepatoma cells are more sensitive than normal hepatocytes. As shown by scanning electron microscopy the volume increase of both types of cells is accompanied by disappearance of microvilli. In contrast to phalloidin poisoning no protrusions were formed when the cells were exposed to leucocidin under isotonic conditions.


Some pharmacological properties of the false cholinergic transmitter acetylpyrrolidinecholine and its precursor pyrrolidinecholine. The acetylchline analogue acetylpyrrolidinecholine as well as the choline analogue pyrrolidinecholine were synthesized and the cholinergic properties of both substances were investigated on the guinea-pig ileum, rat blood pressure and frog rectus abdominis muscle. Acetylpyrrolidinecholine was 3-5 times less potent than acetylcholine on the three preparations tested. The dose-response curves to acetylpyrrolidinecholine were shifted to the right in a parallel manner by atropine and (+)-tubocurarine. The dissociation constants for atropine and (+)-tubocurarine obtained with acetylpyrrolidinecholine as agonist were not different from those obtained with acetylcholine. This indicates that acetylpyrrolidinecholine specifically stimulates muscarine and nicotine receptors. Eserine potentiated the effects of acetylcholine more than those of acetylpyrrolidinecholine. Pyrrolidinecholine was only a weak agonist on the guinea-pig ileum. It caused a rise of rat blood pressure in doses higher than 10 mumol per rat. Neuromuscular transmission of the phrenic nerve-diaphragm preparation of the rat was not impaired during a 150 min incubation period with 1 mM pyrrolidinecholine. It is suggested that the possible formation and release of acetylpyrrolidinecholine as a false cholinergic transmitter does not modify neuromuscular transmission in skeletal muscle.


In vivo blocking of SV40 virus-induced antitumor resistance by Syrian hamster sera from early stages of primary SV40 carcinogenesis. The blocking of specific SV40-induced resistance of hamsters by the sera collected from 12 individual hamsters infected with SV40 when newborn was studied at different periods of primary virus-induced carcinogenesis. The serum samples were collected at the following periods of primary carcinogenesis: during the latent period (60 days after virus inoculation), at the day of primary tumor appearance, and 19-36 and 45-57 days after primary tumor appearance. For detection of the blocking activity of the collected sera the cells of transplantable SV40 test-tumor were pretreated in vitro with these sera and with control normal hamster sera, and then used for challenge in vivo in SV40-immunized and normal hamsters. With the use of such method, as a rule, no blocking activity of the serum samples collected at any time after the tumor appearance was observed. However, the sera obtained from 7 out of 12 of these hamsters during the latent period significantly decreased the resistance index of animals challenged in transplantation test with the serum pretreated tumor cells. Possible explanations of these findings are discussed.


Varicella-zoster antibodies in patients with Hodgkin's disease. In an endeavour to find an explanation for th raised incidence of herpes zoster in hemoblastoses, serum samples of 50 patients with histologically confirmed Hodgkin's disease and 80 control sera of normal persons of similar age groups were tested for the presenchods of indirect hemagglutination (IH), complement fixation (CF), and immunodiffusion (ID). The geometric mean titer of IH antibodies and the rate of positive ID results were significantly higher in the patients' group. The geometric mean titers of CF antibodies did not differ significantly in the two groups. A certain correlation between serological results and the histological type of Hodgkin's disease was found, but it was not statistically significant. No other correlation of serological results with clinical data was ascertained.


Histochemistry of oxidoreductases, enzymatic polymorphism and anaplasia of neuroectodermal tumors. Oxidoreductases were studied histochemically in 162 cases of neuroectodermal tumors. In order of decreasing activity in the cytoplasma these enzymes could be arranged as follows: NADH diaphorase, lactate dehydrogenase, NADPH diaphorase, glutamate dehydrogenase, glucose-6-phosphate dehydrogenase, isocitrate dehydrogenase, succinate dehydrogenase, malate dehydrogenase. The weak activity of Krebs cycle enzymes and the relatively strong activity of other oxidoreductases, particularly of lactate dehydrogenase, permits to conclude that glycolysis prevails over oxidative processes in neuroectodermal tumor cells. But this should not be interpreted as a decrease of the Krebs cycle enzymes in astrocytoma and oligodendroglioma cells as compared with their parent cells because the latter themselves display a weak activity of these enzymes. A real decrease of Krebs cycle enzyme activity was established only for tumors, the parent cells of which are characterized by a strong (in choroid-papillomas) or moderate (in ependymomas) activity of these enzymes. Many neuroectodermal tumors, in particular those of astrocytic origin, demonstrate a certain correlation between the amount of cytoplasm and oxidoreductase activity. This results in enzymatic polymorphism of the tumor tissue. A certain similarity was established of the oxidoreductase activity in tumor cells and in reactive hypertophic astrocytes. This indicates that both tumor cells and reactive astrocytes may in certain conditions utilize similar mechanisms of increased metabolism. The oxidoreductase activity correlates not with the grade of anaplasia but with different directions of anaplasia reflected in different variants of neuroectodermal tumors. The concept "anaplasia" includes not only certain degrees of dedifferentiation of tumor cells but, as it has been shown histochemically, also an increase of metabolic processes in the tumor cell cytoplasma.


A clinico-pathological study on the correlation of immunosuppression and multiple primary malignant neoplasmas. At present it is obvious that the incidence of second malignancies in patients with malignant diseases increases after prolonged treatment with immunosuppressive, or antineoplastic agents. The occurrence of such additional malignant diseases was analysed in our five-years autopsy material. During this period 7670 autopsies were performed. Malignant diseases were observed in 1707 cases (22.1%) and among them in 58 cases were proved multiple primary malignant neoplasms (3.3%). The average time between the occurrence of initial and second tumors was 29 months. The frequency of second tumors in patients with leukemias (mainly chronic lymphocytic leukemia) were four times higher than in patients with tumors of epithelial origin. Hepatocarcinomas arose in cirrhotic livers and astrocytomas were often followed by new malignancies. In consequence of successfully applied surgical, radiological and combined immunosuppressive, antineoplastic therapy the survival of cancer patients lengthened, so among different side-effects of the used therapy the oncogenesis cannot be left out of account. The danger of subsequent malignant tumors makes it imperative that immunosuppression should only be applied when strictly indicated.


The levels of NAD and NADH in blood of patients with cancer. The levels of nicotinamide adenine dinucleotide (NAD) and NADH in blood of patients with cancer in various stages of development as well as their modification after glucose administration were determined in 188 respectively 77 cases. NAD was significantly decreased in patients with cervix and breast carcinoma but unaltered in patients with metastatic cancers (and lung carcinoma) comparatively with healthy subjects. At 48 hours after i.v. administration of glucose, NAD increased significantly in patients with cervix carcinoma but was unaltered in patients with metastatic cancers (an lung carcinoma). NADH in both cases has given little conclusive results.


Nonspecific immunosuppression and expression of avian myeloblastosis virus (BAI strain A). Chickens were treated with cyclophosphamide in order to induce nonspecific immunosuppression. Treated and untreated animals were injected with avian myeloblastosis virus (AMV) or myeloblasts at the age when a pronounced resistance to the disease is observed. Chickens treated with cyclophosphamide and then challenged with AMV developed acute myeloblastic leukemia in 70 percent. Similarly treated chickens transplanted with fresh AMV producing myeloblasts exhibited 30 percent incidence of myeloblastosis. In contrast, the control animals without treatment showed no myeloblastosis either after myeloblasts application or AMV injection. These results have shown that nonspecific immunosuppression by cyclophosphamide treatment strongly affects the expression of AMV in age-resistant chickens.


Infection of human embryo cells with avian sarcoma virus B77 in vitro. Human embryo cells were infected with avian sarcoma virus B77--Hu(B77). The virus genome was detected in the Hu(B77) cells during subsequent cell passages in the uncloned cells as well as in single-cell clones. Despite the presence of the virus genome in cells, the growth properties did not differ from uninfected cells. The Hu(B77) cells did not reveal the transformed phenotype in vitro.


An in vitro study of the oncogenic effects of two variants of avian sarcoma virus B77 on rat embryonal fibroblasts. Two variants of avian sarcoma virus B77 have been tested for their heteroinductive oncogenic in vitro effects on rat embryonal cells. Variant 22-B77V belonging to subgroup B, had been cultured for a long time exclusively on its original host, i.e. on chicks, before being used in the study. This variant of B77V yielded negative results in repeated experiments on rat cells. Variant 55-B77V belonging to subgroup C and which had been made to pass through rat cells, caused a malignant transformation in the latter. The resultant type of this interaction were virus-producing tumorous cells (designated LWF B55). This study present some of the basic characteristics of these cells.


Growth characteristics and tumorigenicity of long-term cultures of the rat RBA myelogenous leukemia. A continuous cell line, RBA-BC, has been derived from the buffy coat of heparinized blood of a Sprague--Dawley rat with developed RBA myeloblastosis. Separations of myeloblasts from the RBA-BC cell line was accomplished by velocity sedimentation in Verografin density gradients. The cells were monitored during long-term culture by generation time analysis. Cytochemical studies revealed the myelogenous type of RBA leukemia.


Possible role of dopamine in diethylstilbestrol-elicited accumulation of cyclic AMP in incubated male rat hypothalamus. Pimozide and haloperidol, the dopaminergic antagonists, have been shown to effectively block diethylstilbestrol (DES)-elicited [3H] adenosine 3', 5'-monophosphate (cAMP) accumulation in incubated hypothalami from immature male rats. Several noradrenergic antagonists (i.e. propranolol, MJ-1999 and phenoxybenzamine) failed to reduce [3H]cAMP levels significantly. Pimozide was found to be more specific than haloperidol as it did not effect norepinephrine (NE)-elicited formation of [3H]cAMP, while the latter blocker caused a 48% reduction of the stimulation by NE. Pimozide was also more potent than haloperidol in inhibiting DES-stimulated [3H]cAMP formation. It can be inferred from these data that as a result of DES-receptor interaction dopamine (DA) is released and in turn activates DA-sensitive adenylate cyclase in the hypothalamus.


The effects of cholinomimetic drugs and atropine on ACTH release. Acetylcholine (1 and 50 mug), carbamylcholine (1 and 10 mug), and oxotremorine (10 mug) were infused into the 3rd ventricle of rats with deafferented medial basal hypothalamus (MBH); infusions failed to stimulate ACTH release as shown by the plasma corticosterone level. Implants of an atropine-fluorescein mixture (200-250 mug) inhibited the stress-induced rise of plasma corticosterone only when dye from the implant stained large portions of the median eminence and the basal hypothalamus. Atropine implants near the electrodes inhibited the rise in plasma corticosterone usually produced by electrical stimulation in the anterior hypothalamus. Atropine crystals or a 2% atropine sulphate solution placed on the median eminence blocked conduction of action potential in the hypothalamo neurohypophyseal and tubero-infundibular pathways. The evidence will be discussed for and against the participation of a cholinergic synapse (in the MBH) that activates ACTH release after stressful neural stimuli.


The temporal and periodic organization of REM eye movements in mental retardation. Time trends and periodic cycles in REM sleep eye movements were examined in 6 functional and 6 mongoloid adolescent retardates. Both groups of subjects showed approximately equal percentages of linear trends, quadratic trends, linear and quadratic trends, and absence of trend. The eye movement time-series were subject to an orthogonal spectral analysis. In both groups of subjects, peak spectral estimates generally occurred at 10.6- or 21.3-min periods. Following this, the eye movements of the longest REM periods (and one long period of pre-sleep wakefulness) were subject to a least-squares spectral analysis. This analysis yielded estimates ranging from 22 to 44 min. Eye movements lacked the ultradian organization previously reported for eye movements of normal subjects. These results imply that the ultradian organization of REM sleep and the ultradian organization of eye movements may be independent.


Extensive calcification in a tumor of the glomus jugulare. Extensive calcification in the intracranial portion of a glomus jugulare tumor is reported. This finding has not been reported previously in the literature. The patient probably was afflicted with this process for many years and presented with a subarachnoid hemorrhage. The combination of this tumor with subarachnoid hemorrhage is rare.


[Controlled trial with ACTH and corticosteroids in patients with cholestatic viral hepatitis]. Comparison was made between synthetic ACTH (tetracosactide), betamethasone and a normal paradigm (dietary management with glucose solutions, detoxicants and vitamins) over a period at least of 21 days in 27 patients with cholostatic viral hepatitis. Assessment of the clinical and laboratory parameters by means of both covariance and multivariance analysis showed that hormonal therapy offered no significant advantages with respect to the basic treatment protocol.


Serum 17 alpha-hydroxyprogesterone in patients with gestational trophoblastic neoplasms. Serum 17alpha-hydroxprogesterone (17-OHP), progesterone (P), and human chorionic gonadotropin (hCG) levels were measured by specific radioimmunoassay in 19 patients undergoing laparoscopy or laparotomy with either unevacuated molar pregnancy or nonmetastatic gestational trophoblastic neoplasms (GTN), in 10 normal pregnant patients at equivalent gestational age (7-21 weeks), and in 4 patients with metastaic GTN following hysterectomy and bilateral salpingo-oophorectomy. All patients with theca lutein cysts had significantly elevated serum 17-OHP levels compared to those in 1) normal pregnancy, 2) patients with GTN and normal-size ovaries, 3) patients with metastatic GTN in the absence of ovaries (P less than 0.02). Levels of serum 17-OHP but not P correlated with the degree of ovarian enlargement (r = 0.87, P less than 0.05). Serum P concentrations in patients with theca lutein cysts, although higher than the levels in cases of GTN with normal-size ovaries, were not significantly different from the levels in normal pregnancy (P greater than 0.05). Serum hCG levels in patients with theca lutein cysts, though higher than the normal pregnancy levels (P less than 0.05), were not significantly different from those in cases of GTN with normal-size ovaries and GTN without ovaries (P greater than 0.05). Under the conditions studied, no correlation was observed between serum hCG and P levels in our cases of GTN. Increased serum 17-OHP level in a patient with GTN suggests the presence of theca lutein cysts.


Effect of treatment with estrogen conjugates on endogenous plasma steroids. Plasma dehydroenpiandrosterone sulfate (DS), androstenedione (A2), estrone plus estradiol (E1+2) and estriol (E3) were measured in women treated with conjugated estrogens and controls. Plasma DS, A2, and E1+2 levels were higher in the premenopausal controls than in the untreated postmenopausal women; E3 levels were below the sensitivity of the assay. Estrogen conjugates, 1.25 mg or 2.5 mg daily, increased plasma E1+2 to levels similar to those in premenopausal untreated women, and E3 became readily assayable. A 0.625-mg dose increased the level of E3 without affecting E1+2; consequently, the plasma E3/E1+2 ratio was elevated. It is concluded that at the lowest dose 16 alpha-hydroxylase activity, perhaps by substrate induction, is sufficient for the exogenous esogenous estrogens to be largely metabolized to E3. Estrogen conjugates had no detectable effect on plasma DS or A2.


Synergistic bacterial gangrene following abdominal hysterectomy. A case of synergistic bacterial gangrene occurring after abdominal hysterectomy in a 43-year-old patient is presented. After multiple debridement procedures and porcine grafts, the patient responded satisfactorily and eventually recovered.


Rare tumors of the orbit during childhood. Multiform glioblastoma, melanoma, cystic lymphangioma, hemangioendothelioma: 67ga scanning and anatomico-pathological aspects. Four cases of orbital tumours observed in pediatric subjects aged 16 months, 2 1/2, 51/2, and 8 years are presented. Stress is laid on the rarity of these tumors: multiform glioblastoma, melanoma, cystic lymphangioma and hemangioendothelioma. The 67Ga citrate-scanning investigations and the anatomopathological findings are described in detail.


Objective signs of inferior alveolar nerve anesthesia in exodontia. Objective determination of the adequacy of inferior alveolar nerve anesthesia is difficult. A dependable objective test based upon clinical observation of hundreds of patients during extraction of mandibular teeth, is presented. Following routine inferior alveolar nerve block anesthesia, time is allowed for subjective signs of "tingling" of the lower lip. Fully aware that the patient may respond to pressure in the absence of pain, the tooth to be extracted is tested. Gradual pressure of increasing intensity is applied with extracting forceps for 3 seconds, then suddenly released, providing a sudden stimulus. If the patient immediately moves or "jumps," this response indicates inadequate anesthesia. If the patient does not respond to the sudden release of pressure, that tooth can be removed painlessly even though the patient may still respond to pressure.


[Guidelines for the treatment of facial-nerve injuries]. The electroneuronographic registration of the summating potentials of the face muscles has been used to assess the prognosis in 72 traumatic facial nerve lesions. The following guide-lines have evolved from the experience gained: (1) a conservative attitude is indicated if the degeneration of the motor fibus is less than 90% 6 days following onset of the palsy and (2) a surgical revision of the facial nerve is indicated if the degeneration of the motor fibres reaches more than 90% during the first 6 days following onset of the palsy. The clinical differentiation in early and delayed traumatic facial palsy cannot be used as a prognostic sign. The amount and rapidity of degeneration of the motor fibres has more prognostic value than the latency between trauma and onset of the palsy. The importance of electroneuronography for the prognosis of iatrogenic facial nerve lesions as well as the value of a careful follow-up of regeneration in facial nerve lesions with preserved continuity of the nerve are discussed.


Corticosterone in serum of adrenalectomized male rats. Following total adrenalectomy of adult male rats, the corticosterone level in the sera declines in the first and second postoperative day from 4.79+/-1.66 mug to 0.56+/-0.24 per 100 ml. On the third postoperative day the value increases to 1.7+/-0.67 mug/100 ml and attains again control level on the 14th p.o. day (5.4+/-3.3 mug/100 ml). Stress, but not 10 i.u. of ACTH causes a further increase of the corticosterone content in the serum reaching 14.3+/-3.5 mug/100 ml, this value representing a much lower amount of hormone in comparison to stressed control animals (36.35+/-2.4 mug/ml). In adrenalectomized and castrated animals no corticosterone could be detected 14 days p.o.; nor do stress or ACTH cause any change.


Lobular carcinoma of the breast in situ and infiltrating. Lobular carcinoma in situ, while histologically delineated almost 35 years ago, is still being diagnosed and reported with differing qualitative and quantitative criteria. It is important to recognize this lesion because of its frequent bilaterality and risk of developing infiltrating carcinoma in the affected breast. Because of the apparent morphologic identity of the cell of ALH and the cell of LCIS, because both are associated with some increased risk of developing carcinoma, and because both have an increased propensity to be found simultaneously with carcinoma--especially lobular carcinoma--we believe these lesions should be looked upon as basically identical. Beginning with Ewing's astute intuition nearly 60 years ago18 and culminating with the 1967 report of McDivitt and others,50 pathologists and surgeons have gradually become aware of the long-term risk associated with this nonobligatorily, premalignant lesion (LCIS). As further studies have become available, it is clear that a large majority of women with lobular carcinoma in situ or atypical lobular hyperplasia retaining the involved breast will not develop infiltrating carcinoma. However, there is an increased risk of roughly 1 percent per year. Because of the increased risk, we believe that a meticulous long-term followup is mandatory, and suggest a physical examination every two to three months for the first few years supplemented with yearly mammography (as is now being done at Memorial Hospital for patients with atypical breast lesions6). With such a followup, we believe the mortality should fall well below 5 percent in a 20-year followup, and believe this is a risk that may be acceptable to the patient and her surgeon. The boundaries of the morphologic changes acceptable as ILC have been difficult for many pathologists (including ourselves) to delineate. This is borne out by widely varying incidence figures and is due, at least in part, to the frequent admixture of ILC with poorly differentiated duct carcinoma as well as problems in deciding how large and pleomorphic the cell nuclei may be, and still be designated as lobular in origin. We believe that about 4 to 6 percent of all breast carcinomas are infiltrating lobular, and incidence rates widely divergent from this should be scrutinized carefully. Prognostically, ILC behaves similarly to poorly differentiated duct carcinomas except for some increased risk of subsequent contralateral carcinoma. Future study of lobular disease should be directed toward its histogenesis, long-term risk, and the relative success of different treatment modalities. Hormonal studies and cell culture with chromosomal analysis might shed some light on the mechanism of what may be postmenopausal regression of LCIS.


Light microscopy of selected viral diseases (morphology of viral inclusion bodies). A significant number of viruses produce intracellular changes, inclusion bodies, which do have specific and consistent features important to their identification. The inclusions must fulfill the designated criteria for the specific virus or a member of that group. The specific features of viral inclusions are consistent fixative-dependent artifacts: uniformity and familiarity of staining quality are important to the consistency of recognition and identification. Bouin's or Zenker's-acetic acid are the fixatives of choice, and a well-done hematoxylin and eosin stain offers the greatest uniformity of quality. Viral inclusions are not static objects in a cell and recognition of the range of development is often more important than the appearance of a single inclusion. Generally, most viruses within a group cause identical intracellular reactions and produce similar inclusion bodies, thus permitting the pathologist an accurate prediction of the etiologic agent in clinically undiagnosed or unsuspected viral disease. By correlating viral inclusion identification, clinical history, and physical findings the anatomic pathologist can make the diagnosis of many viral diseases rapidly, inexpensively, and with a high degree of accuracy when compared to the complex, expensive, and time consuming procedures of animal inoculation, tissue culture, or serology.


The mechanism of urinary concentration in nephrogenic diabetes insipidus. The authors have evaluated urinary adenosine 3',5'-monophosphate (cyclic AMP) excretion and renal function during Pitressin administration, hypertonic saline administration, and water deprivation in two siblings with vasopressin-resistant diabetes insipidus and in normal control subjects. After vasopressin administration normal subjects experienced a 2-fold rise in urinary cyclic AMP excretion from 3.2 +/- 0.7 to 5.6 +/- 1.3 nmol/min (P less than 0.001) whereas cyclic AMP excretion was unchanged in both patients (patient AC 4.4 +/- 0.9 to 4.3 +/- 2.1; patient TC 2.2 +/- 0.9 to 2.6 +/- 0.9 nmol/min) with nephrogenic diabetes insipidus (NDI). Urinary cyclic AMP excretion was measured during infusion of 2.5% saline, after vasopressim administration, and after water deprivation. Cyclic AMP excretion was not different from control values in the NDI patients during any of the experimental conditions. Furthermore, there was no difference in cyclic AMP excretion when periods of dilute urine excretion (patient AC 4.5 +/- 1.1; patient TC 2.1 +/- 0.8 nmol/min) were compared with periods when urine concentration was greater than that of plasma (AC 3.5 +/- 1.3; TC 1.8 +/- 0.9 nmol/min). Both subjects responded to parathyroid hormone infusion with a 2-fold increase in urinary cyclic AMP excretion. Excretion of concentrated urine was paralleled by a marked decrease in urine flow to less than 1 ml/min/m2. During periods of hypotonic urine excretion (Uosm/Posm less than 1.0) average glomerular filtration rate (GFR) in patient AC was 67.0 +/- 3.0 ml/minm2 whereas in patient TC it was 70.1 +/- 8.1 ml/min/m2. When each patient was excreting a hypertonic urine (Uosm/Posm greater than 1.0) after fluid deprivation their GFR had decreased significantly (P = 0.001) to 31.6 +/- 8.9 and 33.3 +/- 10.3 ml/min/m2, respectively. Ability of these two subjects with NDI to concentrate their urine to Uosm/Posm greater than 1.0 in the absence of an increase in urinary cyclic AMP but associated with a decrease in GFR to 50% normal indicates that urinary concentration was effected by a reduction in GFR rather than a partial response to antidiuretic hormone (ADH). Their ability to concentrate their urine during periods of modest volume depletion would protect them from progressing to more severe stages of dehydration and result in the relatively benign course of their disease. It is feasible that in patients previously reported to have had clinically "partial" NDI this mechanism may have been operative.


The renin-angiotensin system and sodium excretion during gestation. The renin-angiotensin system shows a marked increase in activity in normal pregnancy, with the major changes occurring in the first trimester. Evidence is presented to show that there is a physiologically inactive prorenin present in the plasma during pregnancy that achieves highest levels in the first half  of pregnancy. The significance of this "acid-activated" renin has yet to be determined. Plasma angiotensin II levels are elevated throughout normal pregnancy, and a significant positive relationship has been established between maternal venous and cord venous levels at delivery. Recent studies on plasma AII levels in pregnancy hypertension have shown a positive correlation between diastolic blood pressure and plasma AII, and there is evidence that the high circulating levels of angiotensin may result in a feed-back suppression of renal renin release. Studies on the fetal circulation at delivery have also shown high levels of AII in the cord venous blood of infants born to mothers with pregnancy hypertension. Despite the increased level of activity of the renin-angiotensin system in normal pregnancy, sodium homeostasis is maintained. The possible reasons for the increased activity are discussed briefly.


Control of ACh sensitivity in temporarily unconnected ("decentralized") segments of diaphragm-muscle fibres of the rat. 1. After a local lesion of the diaphragm muscle, which produced a segment of innervated muscle fibres connected with an intact nerve-free segment by a region of crushed muscle fibres, the sensitivity to ACh, the presence of tetrodotoxin resistant action potentials (AP) and the transmission of AP along the muscle fibres were studied. 2. Three days after local injury of the diaphragm muscle ACh sensitivity and TTX resistance appeared in the crushed and nerve-free segments between the place of injury and the tendineous attachment. 5-7 days after injury transmission of action potentials through the damaged to the undamaged ("decentralized"), nerve-free part of the fibres is restored. High ACh sensitivity and TTX resistance of the latter segment, however, are completely lost only 20 days after the local injury. During this period contractility of the muscle remains practically unchanged. Enzymatic activities (SDH, ATPase and phosphorylase) of the damaged part were lost 3 days after crushing and recovered slowly between 7-10 days of regeneration of the diaphragm muscle fibres. 3. The experiments suggest that during regeneration of damaged muscle fibres supersensitivity to ACh remains high inspite of normal AP activity and that intracellular mechanisms may be involved in the induction and disappearance of ACh hypersensitivity.


A deoxyadenylate kinase activity associated with polynucleotide phosphorylase from Micrococcus luteus. We report here the presence of two enzymatic activities associated with highly purified preparations of polynucleotide phosphorylase from Micrococcus luteus. The first, a nuclease activity, which is not separated from the phosphorylase on hydroxylapatite, may be due to substitution of H2O for phosphate in the phosphorolysis reaction. The second activity, a deoxyadenylate kinase, the bulk of which is not resolved from the phosphorylase using gel filtration, sucrose density gradient centrifugation, DEAE-Sephadex, or hydroxylapatite chromatography, may represent a new activity of polynucleotide phosphorylase or be due to an enzyme which is tightly bound to the phosphorylase. Several properties of the kinase are described and its possible significance with respect to the overall enzyme mechanism is discussed.


Evolutionary trends in 18S ribosomal RNA nucleotide sequences of rat, mouse, hamster and man. The large T1 ribonuclease fragments of 18S ribosomal RNA from four mammalian species, rat, mouse, hamster and man, were compared by two-dimensional homochromatography fingerprinting. The nucleotide sequences of the large T1 ribonuclease fragments, polypyrimidines and polypurines which were different among the four mammalian species were determined and compared. The method used for determining nucleotide sequences utilizes 32p-labeling of oligonucleotides at their 5'-termini by polynucleotide kinase, partial digestion by ribonucleases and analysis of labeled spots by homochromatography-fingerprinting. Several examples of point mutations were detected. It was of interest that the 18S rRNA of Chinese hamster has more oligonucleotide sequences in common with those of man that rat or mouse.


Partial characterization of an endonuclease activity which appears in nuclease free T4 polynucleotide kinase. A nuclease activity has been found to appear in preparations of T4 induced polynucleotide kinase which had originally been nuclease free. The nuclease introduced random nicks into T7 DNA suggesting that it was an endonuclease. Destabilization of the kinase molecule by osmotic shock or by the removal of reducing agents, ATP or salts was shown to stimulate the endonuclease appearance. The molecular weight was found to be 32,000 +/- 10% by gel filtration on G100 Sephadex. The nuclease was active over a wide pH range from pH 5.0 to pH 9.2 in a number of buffer systems and required MgCl2 and reducing agent for maximum activity. Sodium azide did not affect the nuclease appearance.


A model for replication of the ends of linear chromosomes. Linear chromosomes possessing internal repeats of their terminal sequences can form intramolecular crossed-strand exchanges that allow replication of the chromosome ends. Evidence is discussed that such a mechanism may be utilized during replication of herpes simplex virus DNA and during replication of macronuclear DNA from the hypotrichous ciliate Oxytricha.