Possible role of the 72,000 dalton DNA-binding protein in regulation of adenovirus type 5 early gene expression. Relative abundances of early virus RNA species in the cytoplasm of cells infected with wild-type adenovirus type 5 (WT Ad5) and a temperature-sensitive "early" mutant, H5ts125 (ts125), were compared by hybridization kinetics using separated strands of HindIII restriction endonuclease fragments of Ad5 DNA. 1-beta-D-Arabinofuranosylcytosine (ara-C) was used to limit transcription to early virus genes in cells infected by WT virus. At 40.5 degrees C, a restrictive temperature for ts125, three to seven times as much virus RNA from all four early regions of the genome accumulated in the cytoplasm of cells infected by the mutant as accumulated in cells infected by WT. At 32 degrees C, no such difference in the relative abundances of cytoplasmic virus RNA was observed. The capacity to synthesize a 72,000-dalton (72K) virus polypeptide, presumably the single-stranded DNA-binding protein that is defective in ts125 at restrictive temperatures, was compared in cells infected at 40.5 degrees C in the presence of ara-C with the mutant or WT Ad5. The rate of 72K polypeptide synthesis, measured by sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis of [35S]methionine-labeled polypeptides and autoradiography, was greater at 15 h after infection in ts125-infected cells than in cells infected by WT. A time course experiment showed that the rate of synthesis of the 72K polypeptide increased continuously in ts125-infected cells during the first 15 h of infection, relative to the rate in WT-infected cells. These data are consistent with the hypothesis that Ad5 early gene expression is modulated by the product of an early gene, the 72K DNA-binding protein.


Inhibition of transcription by immunoglobulins directed against the ribonucleoprotein of homotypic and heterotypic vesicular stomatitis viruses. Specific antisera were raised by immunization of rabbits with purified nucleocapsids containing only RNA and N protein (ribonucleoprotein, RNP) obtained from vesicular stomatitis (VS) virions of the Indiana (VSInd) and the New Jersey (VSNJ) serotypes. The specificity of anti-RNPInd serum was demonstrated by selective precipitation of homotypic RNPInd devoid of L and NS proteins; anti-RNPInd serum also selectively precipitated soluble N protein present in cytoplasm of infected cells, but co-precipitated a limited amount of contaminating soluble NS protein. Immunoglobulins prepared from each homotypic antiserum markedly inhibited in vitro transcription of VSInd and VSNJ virions. Anti-RNPInd and anti-RNPNJ immunoglobulins also exhibited cross-reactivity by inhibiting transcription of heterotypic virions, but only to a much lesser degree than in the homotypic reaction. Anti-RNPInd immunoglobulin did not inhibit transcription of the antigenically unrelated Chandipura rhabdovirus, but anti-RNPNJ immunoglobulin did to a very limited extent. The transcription inhibitory activity of anti-RNPInd immunoglobulin was not dependent on RNP immunoprecipitation activity, which could be diluted out well before loss of antitranscriptase activity. Anti-RNPind immunoglobulin appeared to exert its effect on transcription by blocking elongation rather than initiation or reinitiation of RNA transcripts.


In vitro transcription of mouse mammary tumor virus RNA into DNA. The in vitro transcription of the B-type mouse mammary tumor virus RNA into DNA is similar to that obtained with certain C-type viruses: synthesis can be initiated on an RNA primer molecule approximately the size of a tRNA, and a unique linkage (rA-dC) exists between this primer and the DNA product.


In vitro termination of adenovirus DNA synthesis by a soluble replication complex. The double-stranded DNA from a soluble DNA replication complex that was labeled with deoxyribonucleoside triphosphates and completed in vitro was digested with EcoRI, Sma I, and Hpa I restriction endonucleases. All regions of the adenovirus type 2 genome were labeled in vitro, but restriction fragments derived from the ends of the DNA molecules were relatively more highly labeled than those derived from internal regions. The in vitro endogenous DNA polymerase reaction also exhibited strand-specific labeling near the molecular ends, in that restriciton fragments from the left end were labeled predominantly in the r strand and fragments from the right end were labeled predominantly in the l strand.


Laetrile toxicity studies in dogs.

Laetrile toxicity studies in dogs. Dogs were fed laetrile and fresh, sweet almonds under various conditions. The doses of laetrile were similar to those prescribed for patients with cancer and ranged on a basis of gram to square meter from an equivalent of the oral dose for man to five times this dose. Six of the ten dogs died of cyanide poisoning. One dog recovered, and three dogs, at the time of sacrifice, demonstrated various levels of neurologic impairment, ranging from difficulty in walking to coma. These studies demonstrate that oral laetrile is highly toxic when taken with some common table foods. We predict that there will be an increased incidence of cyanide poisoning in man as laetrile becomes more readily available.


Diabetes mellitus following rodenticide ingestion in man. Ketotic, insulin-requiring diabetes mellitus and a severe peripheral neuropathy developed in a previously healthy 25-year-old man several days after he attempted suicide with rat poison containing N-3-pyridylmethyl N'-p-nitrophenyl urea. Study of islet-cell function ten months after ingestion showed a reduced disappearance rate of intravenous glucose and depressed C-peptide response to intravenous glucose when compared with a normal control but no impairment of glucagon release after intravenous arginine stimulation. Nerve conduction studies demonstrated severe sensory and mild motor neuropathy. Quadriceps capillary basement membrane thickness was in the diabetic range. Because at least 15 similar occurrences have been reported to the manufacturer, this agent appears to be diabetogenic in man, probably causing beta-cell destruction. Niacinamide, which can prevent glucose intolerance in both streptozocin- and alloxan-treated animals and prevents death in rats given this rodenticide, may be a useful antidote.


Hepatic tumors, surgical treatment and its results. During the period from March 1954 to June 1976, 489 cases of hepatic tumors were experienced at the Department of Surgery, National Taiwan University Hospital. The pathological classification are: Hemangioma 5; Cystic hamartoma 2; Hemangioendothelioma 2; Non-parasitic localized cyst 5; Polycystic liver 9; Tuberculoma 1; Actinomycosis 1; Hyperplasia 1; Adenoma 1; Primary liver sarcoma 2; Primary liver cancer 442; and Metastatic liver cancer 18. Of the 489 hepatic tumors, two cases were treated with excision, 148 cases with hepatic lobectomy and nine cases of polycystic liver were treated with fenestration. Benign hepatic tumors are all well up to eight years after the operation, 14 cases of primary liver cancer survived five to 17 years after hepatic lobectomy.


Tumor embolism in the right atrium after hepatic artery ligation for hepatoma. A case of tumor embolism in the right atrium after hepatic artery ligation for hepatoma was reported. Patient was a 55 year old male complaining of upper abdominal pain. Preoperative angiogram and scintigram revealed multinodular hepatomas in the right and middle his condition gradually deteriorated thereafter and sudden hypotension occurred on the 22nd postoperative day with a fatal sequela. At autopsy, the orifice of the right atrium was impacted by a tumor embolus. The cause of death seemed to be attributable to this embolus liberated from the hepatic veins. Accordingly, we emphasize the necessity of careful angiographic evaluation not only of the hepatic inflow but of the outflow tract in performing hepatic artery ligation as a treatment of nonresectable hepatoma.


[Microcirculatory hemostasis and its changes in cardiovascular diseases]. The indices of microcirculatory hemostasis were studied in 955 persons, 200 of whom were healthy while 755 had diseases of the heart and vessels. The results were compared with blood fibrinolytic and coagulation activity and with the functional condition of the basophil-mast cell apparatus. A reverse trend in the functional shifts of microcirculatory and plasmo-coagulative hemostasis in the reaction of emergency adaptation was established. Delayed formation and increased resistance of platelet aggregates with simultaneous decrease in the number of platelets in circulation were revealed in cardiovascular diseases during a remission, which reflects intensified expenditure of the platelets or their aggregation energy in the microcirculatory channel. The administration of exogenous heparin, just like the period of basophil degranulation in stresses of various origin, is attended with distinct platelet hyperfunction which allows small doses of heparin to be used for the prevention of diapedetic hemorrhages in hypertensive disease.


[Diagnosis and treatment of ocular myasthenia (author's transl)]. After discussion of the modern concepts of pathophysiology of ocular myasthenia the ocular symptoms such as ptosis and eye muscle palsies are discussed. As important diagnostic sign the Simpson lid fatigue test before and after application of Tensilon is described. For diagnosis of myasthenic eye muscle palsies electrooculography has a special significance especially in connection with the application of Edrophonium, which normalizes myasthenic hypometric saccades and transforms them even in hypermetric saccades. In doubtful cases of eye muscle palsies the electromyogram of the affected muscle in connection with the Edrophonium-test is extremely valuable. With regard to modern treatment apart from cholinesterase inhibitors (Pyridostigmine, Neostigmine) thymectomy, the application of corticosteroids, ACTH and especially also immune suppressive drugs (Imurel etc.) is discussed. Of great significance in ocular myasthenia is the local application of cholinesterase inhibitors like Eserine, Prostigmin or Phospholine Iodide.


[Regulation of the human erythropoietic proliferation pool by the microenvironment (author's transl)]. The Effects of Various Substances in the Culture Microenvironment on the Proliferation of Erythroblasts in vitro were Studied and Evaluated by differential cell and mitosis counting in normal and pathologic bone marrow. The mitotic frequency and maturation correlated directly with the erythropoietin content of the medium and were potentiated on addition of either folic acid, etiocholanolon or cAMP. However, erythroblast proliferation was stimulated independently of erythropoietin concentration when either cobalt, and androgen or an anabolic substance was added to the medium. An iron-deficient medium prevented the maturation of erythroblasts to reticulocytes, thereby rendering erythropoiesis ineffective. The existence of an erythrocyte chalone ineffective. The existence of an erythrocyte chalone or an erythropoietin inhibitor could not be deduced from our experiments since the transformation of pluripotent to erythropoetin-sensitive stem cells is not included. A stimulation of the granulopoietic proliferation pool occurred when serum from patients with either polycythemia vera or after acute blood loss was added to the medium.


Pox infection in white rats.

Pox infection in white rats. 2 pox outbreaks among white rats in a breeding colony are described. The infection occurred in 3 different forms: pulmonary, dermal and mixed. Apparently healthy animals appeared to be virus carriers. The virus isolated belonged to the genus Orthopoxvirus of Poxviridae family, and was very close to cowpox virus. It differed from reference strains of cowpox virus in having a lower ceiling temperature and a higher pathogenicity for white rats.


Effects of mercury on lysosomal protein digestion in the kidney proximal tubule. The effect of mercury on renal lysosomal protein digestion was studied after administration of mercury in vitro and in vivo. Mercuric chloride or methylmercury chloride was added in vitro to lysosomal enzymes isolated from normal rats, and subsequently, digestion experiments were carried out using 125I-labeled lysozyme or cytochrome c as substrate proteins. Both mercury compounds produced a concentration-dependent inhibition of the degradation of the proteins, mercuric chloride being the strongest inhibitor. Mercuric chloride was also administered to rats in vivo for 5 to 8 months. Renal lysosomal enzymes from these animals also had a decreased ability to digest for the two substrate proteins. Furthermore, the digestion of lysozyme intravenously injected into mercury-intoxicated rats was decreased in renal cortical slices incubated in vitro. Electron microscope autoradiography showed that intravenously injected labeled lysozyme was located primarily over lysosomes in proximal tubule cells 1 hour after injection in both control animals and mercury-intoxicated rats. These results suggest a decreased catabolism of low molecular weight proteins in the kidney during chronic mercury intoxication.


Levels of plasma cyclic AMP and insulin in cardiac surgery. Cyclic AMP is a common second messenger for a variety of hormones such as catecholamine, glucagon, and growth hormone, which are affected by cardiac surgery. Changes in plasma cyclic AMP level may thus reflect an altered hormonal milieu. The effects of open-heart surgery on plasma cyclic AMP and its relation with serum insulin were studied in 33 adult patients who underwent cardiac surgery with cardiopulmonary bypass. Plasma cyclic AMP levels were markedly elevated during cardiopulmonary bypass and returned toward normal within several days after the operation. The serum insulin concentration remained low, and no positive correlation was found with plasma cyclic AMP level. The responses were similar in patients who had aorta-coronary bypass grafts and those who had valve replacements.


[Fibrous histiocytomas of the nose and paranasal sinuses (author's transl)]. Fibrous histiocytomas of the nose and paranasal sinuses are very unusual. Two cases of these tumors are reported. Prognostic histological features and biological behaviour are discussed.


Visualization of primate high density lipoproteins isolated by density gradient ultracentrifugation. High density lipoproteins (HDL) isolated by density gradient ultracentrifugation from the plasma of monkeys ingesting semipurified diets are barely visible. This creates difficulty in separating HDL from other lower density lipoproteins following centrifugation and necessitates collecting lared quantities of background density solution to insure complete recovery of the HDL fraction. A simple procedure is described involving the addition of beta-carotene to nonhuman primate plasma prior to centrifugation which results in the delineation of HDL as a discrete yellow-orange band without affecting certain physical properties of HDL or interfering with standard lipid and protein assays.


[Morphological and histochemical changes in the rat liver in chronic furfural poisoning]. The experiment was carried out on rats, which were divided into three experimental and one control groups. The experimental animals were intraperitoneally injected with furfural in the dose of 58 mg/kg body weight for 30 days. In the liver samples obtained at autopsy, apart from routine staining with hematoxylin and eosin, estimation of the activity of the following enzymes was made: succinic dehydrogenase. NADH-tetrazol reductase, lactic dehydrogenase, glucose-6-phosphate, adenosine-triphosphatase, Ca-formol, glucose-6-phosphatase and acid phosphatase. Glycogen content was also evaluated. A temporary decrease in the activity of reactions for the enzymes of tissue respiration, an increase in the activity of glucose-6-phosphatase with a simultaneous decrease of glycogen content, activation of intracellular digestive processes, and inhibition of active transport through biological membranes were found in animals intoxicated with furfural.


Effect of starvation on hepatic glycogen metabolism and glucose homeostasis. The effects of starvation on the hepatic glycogen synthase and phosphporylase systems were sequentially assessed in fed and 24-120-hr-fasted rats. Enzymic changes before and after glucose were correlated with simultaneous measurements of hepatic cyclic AMP and glycogen concentrations and glucose, insulin, and glucagon concentrations in the portal vein plasma. Fasting caused parallel changes in plasma glucose and hepatic glycogen concentrations with decreases by 24 hr and subsequent increases, which correlated with increases in hepatic synthase l and decreases in phosphorylase activites. Hepatic cyclic AMP levels increased as 24-48 hr, decreased below fed levels at 96 hr, and increased again at 120 hr. Fasting caused progressive impairment of glucose disposal, decreased basal and postglucose insulin concentrations, and decreased basal glucagon levels at 48-72 hr. Hepatic synthase l increments following glucose were exaggerated in 48-120-hr-fasted rats, although consistent phosphorylase decrements were seen only in fed rats. There was no clearcut relationship between synthase activation and phosphorylase inactivation following glucose in fed or fasted rats.


Thermolabile repression of cephalosporinase synthesis in Citrobacter freundii. An unusual regulatory system of cephalosporinase synthesis in Citrobacter freundii has been found. When the bacteria are grown at 20 C, the cephalosporinase is synthesized as a typical inducible enzyme and benzylpenicillin acts as an effective inducer. The enzyme, however, is synthesized in the absence of the inducer at growth temperatures above 25 C. when the growth temperature is shifted from 20 C to 37 C, the induction of enzyme synthesis is observed after about one half of the organism doubling time, but it does not occur in the presence of chloramphenicol. The reverse control mutants, the enzyme constitutive synthesis of which is markedly depressed by benzylpenicillin, were isolated from the C. freundii wild strain. The possibility that the enzyme synthesis is governed by a regulatory system analogous to the its mutant of the lac operon in Escherichia coli was suggested.


Rapidly progressive fatal silicosis in a young man. A case is reported of rapidly progressive silicosis in a 30-year-old male who developed symptoms two years after first exposure and died 30 months later. The causative agent was silica powder. This case serves to highlight the fact that this potentially dangerous material is currently being used in industry and that adequate precautions are not always taken.


[Combination or monotherapy of hyperlipoproteinemia typus IIb, IV, V with clofibrate and m-inositolnicotinate or clofibrinic acid (author's transl)]. Clofibrate and m-Inositolnicotinate in a daily dosage of 1,5 g Clofibrate and 1,2 g m-Inositolnicotinate during long term treatment effected a good triglyceride fall in all three lipoprotein fractions, especially in VLDL. The changing of the hyperlipoproteinemia phenotypus IV to the phenotypus IIb or IIa under monotherapy with Clofibrate or clofibrinic acid could not be avoided by the combination of Clofibrate with m-Inositolnicotinate. Approximately every fourth hyperlipoproteinemia phenotypus IV or V, treated with a combination therapy, had an increase of beta-Cholesterol in the pathological range of more than 210 mg/dl. The combination of Clofibrate with nicotinic acid is not of greater value than the monotherapy with Clofibrate (Atromid S). A combination of Clofibrate and nicotinic acid is useful when pure nicotinic acid (3 to 4g/daily) or its esters, for example m-Inositol-nicotinate (4 to 6g/daily) or beta-Pyridylcarbinol (800 to 1200 mg/daily) are administered in therapeutic doses. With the increased dosage of nicotinic acid, one must expect a large "drop out" of these appropriate hyperlipoproteinemia patients because of the side-effects resulting from this particular treatment with nicotinic acid.


RD cells in the laboratory diagnosis of enteroviruses. It is shown that, in addition to the already known replication of Coxsackie A viruses, 16 out of 18 different types of ECHO viruses tested as well as all three types of polioviruses can also be grown in RD cells with the development of a pronounced CPE. One of the exceptions, ECHO virus type 16, can be propagated in either HEL or HFDK cells and the other, ECHO virus type 22, in Vero cells. These four permanent or semi-permanent cell cultures thus prove very useful for the isolation of most of the diagnostically important enteroviruses as well as for their serological identification by neutralization in a micro-titer system described in this paper.


[Hepatic disease in childhood and cytomegalovirus infection (author's transl)]. 135 children with hepatitis and 179 patients with hepatomegaly of undetermined etiology were investigated for complement fixing antibodies to cytomegalovirus. 69 patients with hepatitis (51%) and 74 patients with hepatomegaly (41%) had a titer of 1:4 or above as compared with a control group of children without hepatic pathology in whom positive response occurred in only 20 resp. 18%. The mean titer found in the sero-positive patients was also significantly higher than that in the control group. In contrast, the incidence of positive sera in newborns with hyperbilirubinemia of unknown etiology did not differ significantly from that in a healthy control collective. Of 57 patients with liver disease, 20 excreted cytomegalovirus which was isolated 55 times from 103 urine specimens. These patients included 9 with hepatitis and 11 with hepatomegaly. Of 129 children without any indication of liver disease, cytomegalovirus could be demonstrated in only 3. Our results point toward a causal connection between the presence of cytomegalovirus and the development of hepatic disease in childhood.


EMS-induced dominant lethal dose response curve in DBA/1J male mice. An ethyl methanesulfonate (EMS) induced dominant lethal dose response experiment was conducted in strain DBA/1J male mice. Two methods of scoring for dominant lethals, the classic method (dissecting females at mid-pregnancy) and an alternative method (inspection of uterine scars after litters were weaned) were compared. Results indicate that strain DBA/1J has a similar sensitivity to EMS-induced dominant lethals as has been previously reported for other mouse genotypes. Of the two methods used to score dominant lethals, the classic method is more sensitive.


Induction of dominant lethals with ethyl methane-sulfonate in male germ cells of mulberry silkwork, Bombyx mori l. Sensitivity of male germ cells in the mulberry silkworm, Bombyx mori L., to ethyl methanesulfonate (EMS) was determined by treating newly emerged 5th- instar larvae, and 2-day- and 7-day-old pupae with 3 concentrations, 0.05, 0.1 and 0.15%, of the mutagen. The frequency of dominant-lethal mutations induced by EMS treatment was used as the parameter for the study. Spermatids and spermatozoa were markedly sensitive to EMS. Statistical analysis confirmed that differences in respect of percentage of egg hatch among the 3 different treatments as well as the interactions between the 3 factors, e.g. stages, hatchability and EMS treatment, were highly significant.


Inhibition of angiotensin-converting enzyme for diagnosis of renal-artery stenosis. To determine its utility as an aid in diagnosis of renovascular hypertension, we administered nonapeptide converting-enzyme inhibitor (CEI) (which inhibits conversion of angiotensin I to angiotensin II) (0.25 mg per kilogram) to 14 unselected hypertensive patients undergoing bilateral renal-vein catheterization. In seven (Group I) predominantly unilateral disease was discovered by angiography (renal-artery stenosis in six and hydronephrosis in one); in the remaining seven (Group II) no rennal-artery abnormality was found. In Group I, mean (+/- S.E.) ratio of involved to uninvolved renal-vein plasma renin activity (PRA) increased from 2.94 +/- 0.91 before to 8.36 +/- 2.94 after CEI (P less than 0.01). In Group II, the ratio (of the initially higher to the lower side) was 1.99 +/- 0.49 before and 1.17 +/- 0.07 after CEI (P greater 0.02). Post-CEI PRA was predicted by pretreatment PRA. Mean blood pressure fell in both groups after CEI, and the decrement was predicted by pre-CEI PRA. These data suggest that CEI can be of use at the time of renal-vein catheterization, serving to increase diagnostic accuracy by increasing the difference in PRA between the two sides when there is unilateral disease.


Cushing's disease. Selective trans-sphenoidal resection of pituitary microadenomas. We undertook trans-sphenoidal microsurgical pituitary exploration in 20 consecutive patients with Cushing's disease, eight of whom had normal sellar polytomography. Pituitary adenomas were selectively resected in 17 and histologically confirmed in 14. In one patient total hypophysectomy revealed a 1.5-mm basophilic adenoma, and in two patients vascular anomalies prevented sellar exploration. Hypercortisolism was corrected in 17 patients (i.e., in 16 of the 17 undergoing selective tumor removal and in the one with total hypophysectomy). Panhypopituitarism occurred only in this patient, and transient diabetes insipidus occurred in five. Most patients became glucocorticoid deficient and required replacement therapy. We conclude that pituitary tumors are present in the great majority of patient with Cushing's disease, even in the absence of demonstrable tomographic changes in the sella turcica, and that selective removal corrects hypercortisolism with little morbidity.


Differences in pathogenicity and antigenicity among hog cholera virus strains. Using antiserum against a particular strain of bovine viral diarrhea virus, the strains of hog cholera virus were divided into two groups, H and B, on the basis of the difference in the degree of neutralization. Group H consisted of strains reacting poorly in neutralization, and group B Consisted of strains reacting well with bovine viral diarrhea antiserum. Most of the strains of group H induced a typical clinical form of hog cholera in experimentally infected pigs. Inoculation of pigs with a strain of group B, however, resulted in a chronic type of illness. When immunized with bovine viral diarrhea virus, pigs succumbed to challenge with group H virus after showing clinical signs of hog cholera, but survived challenge with group B virus without manifesting any clinical sign.


Pathological changes in chickens inoculated with reticuloendotheliosis-virus-contaminated Marek's disease vaccine. A disease characterized by delayed growth, anemia, abnormal feathers, and leg paralysis occurred among chickens inoculated with Marek's disease vaccine over a period from spring to fall in 1974. These chickens were recognized among flocks inoculated with the vaccine produced by two vaccine makers. The affected ones were examined pathologically. Gross examination revealed a slight enlargement of peripheral nerves and atrophy of the spleen, thymus, and bursa of Fabricius. Histopathologically, the peripheral nerves had a mild cell infiltration of lymphoid and plasma cells, edema, degeneration of nerve fibers with Schwann's cell proliferation. Perivascular cuffings consisting mainly of lymphoid cells were seen in the brain and spinal cord. Atrophic changes displayed by prominent reduction of lymphocytes were recognized in the spleen, thymus, and bursa of Fabricius. Etiological examination suggested that most of the chickens examined might have been infected with reticuloendotheliosis virus and not with Marek's disease virus. The pathological changes observed in the peripheral nerves and central nervous system, however, were not distinguishable from those of Marek's disease.


Somatosensory-evoked potentials elecited by bilateral stimulation of the median nerve and its clinical application. Somatosensory evoked potentials (SEPs) elicited by bilaterally simultaneous median nerve stimulation (BS) were studied in 28 normal subjects and in 20 patients with various cerebral lesions. In normals, SEP'S evoked by BS were symmetric between the homologous areas of the two hemispheres and resembled the response contralateral to unilateral stimulation (US). In patients with a cerebral lesion, BS responses were not only asymmetric but often differed in wave form from the response of the affected hemisphere to contralateral US. These alterations were attributed to the cumulative interactions of the anomalous contralateral and ipsilateral components. The additional use of BS made it possible to demonstrate SEP abnormalities that might have been regarded as normal or equivocal if only the US method had been used. The neurophysiologic mechanisms of the origin of SEP are discussed.


Serum antibodies to cytomegalovirus in myasthenia gravis: effects of thymectomy and steroids. The etiology of the immunologic abnormalities in myasthenia gravis (MG) remains unknown. Evidence for persistent viral infection was sought by determining serum antibody titers to several enveloped RNA and DNA viruses. Compared to healthy controls matched for age, sex, geography and socioeconomic status, patients with MG were more likely to have elevated titers of complement-fixing antibody to cytomegalovirus (CMV). Patients with MG not treated with thymectomy or steroids had elevated CMV titers, whereas thymectomized or steroid-treated patients did not; the differences were statistically significant (p less than 0.01). These results are consistent with the hypothesis that there is persistent viral antigenic stimulation in the myasthenic thymus, arising from viral protein incorporation into epithelioid-myoid cell surface membranes and subsequent induction of an antibody to these acetylcholine receptor (AChR)-bearing thymic cells. This antibody then cross-reacts with AChR at the neuromuscular junction.


Metastatic oat cell carcinoma simulating liver cirrhosis. A case of oat cell bronchial carcinoma that clinically simulated liver cirrhosis is reported. Post mortem findings included the very unusual intrasinusoidal pattern of liver metastases.


Effects of ritodrine and isoxsuprine with and without dexamethasone during late pregnancy. beta-Adrenergic agents are used to inhibit preterm labor and glucocorticoids to accelerate fetal pulmonary maturation. A study was designed to investigate the metabolic effects of intravenous infusion of ritodrine (150 to 100 microgram/min) or isoxsuprine (200 to 150 microgram/min) in a series of 28 patients with gestations of 28 to 40 weeks, with and without concomitant dexamethasone therapy. Ritodrine was more potent than isoxsuprine in increasing the circulating levels of cyclic AMP, glucose, insulin, and triglycerides. The diabetogenic effect of both ritodrine and isoxsuprine was so slight that it did not have any clinical significance in women with normal glucose tolerance. The results were similar when these beta-adrenergic tocolytics were given to women concomitantly with intramuscular dexamethasone therapy, although dexamethasone appeared to minimally impair carbohydrate metabolism. Both ritodrine and isoxsuprine caused a significant fall in serum iron and potassium, and this effect was unaltered by dexamethasone. Serial serum potassium levels should be obtained during long-term infusion of beta-mimetics.


Arrhenoblastoma during pregnancy.

Arrhenoblastoma during pregnancy. Arrhenoblastoma is a rare complication of pregnancy. A case of fatal nonvirilizing arrhenoblastoma in a pregnant adolescent, the third such case to appear in the literature, is discussed. Fifteen cases of arrhenoblastoma in pregnancy that have appeared in the recent literature are reviewed; 87% of these tumors were virilizing. When virilization is absent the diagnosis is difficult. Among the more common presenting symptoms are weight loss, nausea and vomiting, abdominal pain, and a palpable abdominal mass. Arrhenoblastoma occurring during pregnancy has a 31% maternal mortality, 44% rate of malignancy, and a 50% perinatal mortality.


Therapy of small cell carcinoma of the lung with hexamethylmelamine. Hexamethylmelamine (HXM) is one of the substituted melamines derived from cyanuric chloride [1]. Previous studies with HXM have shown activity against small cell carcinoma of the lung, when it was given as a single agent [2, 3]. Because of relatively mild bone marrow toxicity, HXM was subsequently given in combination with other drugs and/or radiation therapy, in order to improve the therapeutic results. In this paper, our experience with HXM in treatment of small cell carcinoma of the lung is summarised.


Methyl (5-(2-thienylcarbonyl)-1H-benzimidazole-2-yl) carbamate, (R17934), a synthetic microtubule inhibitor, prevents malignant invasion in vitro. The effect of malignant invasion of methyl (5-(2-thienylcarbonyl)-1H-benzimidazole-2-yl) carbamate, (R17934), a microtubule inhibitor, was examined in organotypical co-cultures of mouse sarcoma virus transformed cells (MO4) and embryonic chick tissues. In contrast with control cultures, 1 microgram/ml R17934 prevented invasion of MO4 cells into the host tissue. Since mitostatic doses of 5-fluorouracil did not inhibit invasion, we presumed that the anti-invasive properties of R17934 were correlated with a loss of microtubule mediated directional cell migration.


[Effect of adrenocorticotropic hormone on nucleic acid and protein synthesis and content in the brain of rat embryos in the early stages of neurogenesis]. The injection of adrenocorticotropic hormone (ACTH) of prolonged effect at the doses of 4 and 10 M. U. to the intact rats from the 11th till the 15th day of pregnancy resulted in the twofold increase of protein content in the brain and its decrease in the liver of 15 days old embryos, as compared with the control ones. The content of DNA, RNA and proteins in the placenta of experimental animals increased as well. The rate of incorporation of 3H-thymidine in the liver DNA and 14C-leucine in the liver and brain acid-soluble protein decreased within small intervals of time following the treatment. The total radioactivity of proteins in the liver, brain and placenta calculated per DNA unit was similar to the control one whereas the specific radioactivity of total protein in the liver of experimental embryos was higher than in the control.


[A study of pancreatic insulinomas by electron microscopy. 9 cases (author's transl)]. Nine hypoglycaemia-inducing pancreatic tumours were studied by electron microscopy. In 8 of these tumours, it was possible to identify, within the cell cytoplasm, secretory granules with a "paracrystalline" content, identical in appearance to the granules of the B cell of the normal human pancreas. Thus electron microscopy would appear to be a reliable and sensitive method for the morphological identification of these endocrine tumours of the pancreas.


Lung cancer: the pathologist's role in management. The prognosis of lung cancer is poor, although there is hope for improvement based on early detection through cytologic screening and the use of newer treatment protocols. Although there are difficulties inherent in classifying tumors by type on the basis of very small samples of lesions that are not necessarily homogeneous, the pathologist should use a standard classification that sets forth definite criteria for the differentiated types. Among the common tumors, a small, well-differentiated epidermoid carcinoma with no evidence of lymph node involvement has the most favorable prognosis, and a small-cell anaplastic or oat cell carcinoma the worst. Results of therapy for tumors of other types show much variation. Future studies of lung cancer should include careful clinical staging and, when resection is done, pathologic staging. More study of the immunomorphology of lymph nodes is needed, as this may provide highly useful prognostic information.


Extrapulmonary manifestations of tumors of the lung. Lung tumors, among other, secrete substances which are identical to or which mimic the activities of conventional hormones. These ectopic tumor secretions appear to be anarchistic and beyond the control of mechanisms regulating normal endocrine physiology. Despite the relative inefficiency of these tumors as endocrine organs, production of sufficient quantities of the substance, exceeding those from normal sources, results in recognizable ectopic endocrinopathies. The association of certain tumor cell types with specific hormonal patterns (Cushing syndrome with oat cell; hypercalcemia with squamous cell) is considered consistent with the concept of genetic derepression.


Benign and malignant liver neoplasms: some new etiologic associations. Several agents have recently been implicated in the development of benign and malignant liver neoplasms. Recognition of their possible role by the primary care physician may prevent serious consequences to patients exposed to these agents, which include oral contraceptives, androgens, hepatitis virus, and vinyl chloride.


Perianal Paget's disease.

Perianal Paget's disease. Paget's disease involves the ducts of apocrine glands and the overlying epidermis and is considered to be locally neoplastic. It is capable of developing into invasive carcinoma and in this event the prognosis is poor. The perianal skin may be affected by Paget's disease. This is a rare disorder and the four patients who have been seen with this disease at St Mark's Hospital since 1930 are described.


Effect of infectious bursal disease on the severity of Eimeria tenella infections in broiler chicks. A study was initiated to determine whether prior exposure to infectious bursal disease virus (IBDV) influenced the susceptibility of young broiler chicks to Eimeria tentella infections. When one-day-old chicks infected with IBDV were subsequently challenged with E. tenella at 7 days of age, they suffered significantly higher mortality and lesion scores than their hatchmates which were not exposed to IBDV. Initial exposure to IBDV also had an effect on the development of E. tenella induced hemorrhaging. Hemorrhaging commenced late on the third day following E. tenella inoculation in chicks infected with both IBDV and E. tenella, while bleeding did not occur in the E. tenella alone group until late on the fourth day post-coccidial challenge. No significant differences in weight gains at 7 or 14 days after E. tenella infection were observed between chicks receiving the double infection and those given only E. tenella.


The turnover rate of lipoprotein of d less than 1.006 from plasma of laying turkey hens. The turnover rate of plasma "lower density fraction" (LDF, d less than 1.006) lipoprotein was determined in two laying turkey hens. LDF was labeled by injecting 1-14C-palmitate which was incorporated into the triglyceride (TG) of circulating LDF. The turnover rates were determined from concentration and half life (t1/2) of the ultracentrifugally-isolated LDF. These values were 22 mu equiv. of TG fatty acid (TGFA) per ml. plasma and 431 min. The turnover rate of LDF was 17.57 mili equiv. TGFA/day or 5.0 g. of TGFA/day as sterate. It was estimated that 3.2 g. of LDF-TGFA/day were removed as egg yolk.


The influence of crude cottonseed oil in the feed on the blood and egg yolk lipoproteins of laying hens. Lipovitellin, very low density lipoproteins (VLDL), low density lipoproteins (LDL), high density lipoproteins (HDL), and proteins of d greater than 1.20 were isolated from blood plasma and egg yolks obtained from hens fed a normal diet or one containing 2.5% of crude cottonseed oil. The amounts and compositions of each fraction were determined. Hen blood plasma and egg yolk VLDL and LDL obtained from hens fed a normal diet contained similar levels of lipid, and the fatty acid compositions of those lipids were, for the most part, similar. The percentages of VLDL and LDL in total lipoproteins were similar for plasma and egg yolk obtained from hens fed the normal diet. Separation of VLDL from LDL was not clear-cut in eggs from hens fed the diet which contained crude cottonseed oil. Lipovitellin, as it is isolated from egg yolk, did not appear to be present in any appreciable amount in hen blood plasma. Hen plasma appeared to contain about 10% of a liproprotein d greater than 1.20, the lipid of which was similar in fatty acid composition to that of lipovitellin except for palmitic and oleic acids. Lipids of all the different lipoproteins isolated from plasma and egg yolks of hens fed diets which contained 2.5% of crude cottonseed oil contained more stearic acid and less palmitoleic and oleic acids than did those from normal hens. The increased content of stearic acid increased the density of the lipoproteins so that a larger proportion of the lipoproteins were in the LDL and a smaller proportion were in the VLDL than in lipoproteins from normal plasma and eggs.


Interrelationship of dietary vitamin D3 with zinc and iron in young turkeys. Three experiments were conducted with Large White turkeys to four weeks of age. The interrelationship of zinc and vitamin D3 was studied in two experiments. Iron and vitamin D3 were involved in one experiment. One group of ten male and one group of ten female poults were randomly assigned to each treatment within an experiment. All experiments involved a factorial arrangement of two dietary variables, D3 and either zinc or iron. For example, all possible combinations of four zinc levels; 31, 46, 76 and 106 p.p.m. and three vitamin D3 levels; 600, 1200, and 3600 I.C. units/kg. were fed in the second zinc experiment. Significant (P less than 0.05) weight gain differences occurred among both zinc and D3 levels in this experiment. Four week body weights were 356, 436, 459 and 444 g., respectively, for Zn levels; and 409, 410 and 452 g., respectively, for D3 levels. Significant (P less than 0.05) interactions occurred between dietary zinc and D3 levels, and between iron and D3 levels relative to body weight gain, Vitamin D3 tended to increase tibia zinc levels in poults receiving higher zinc levels; the opposite occurred with lower levels of the element. Zinc level in feces was decreased with vitamin D3. Hemoglobin level was not influenced by D3 but was increased by iron.


Genetic resistance to lymphoid tumor transplants. Day-old chicks of Ottawa strains 4 and 5 and Cornell strains K and S, with or without maternal antibody against antigens related to Marek's disease (MD), were challenged with serial dilutions of MD tumor transplants (JMV-L or JMV-H) or transmissible lymphoid tumor (TLT) of Olson in three consecutive experiments. Thirty chicks were tested in most combinations of strain, dilution and antibody status for a total of approximately 2000 chicks per experiment. Two subsequent experiments, with a total of more than 1900 chickens, investigated the influence of age at challenge on the resistance of strains 4, K and S to JMV-L and the objective of the sixth experiment was to determine the chromosomal sex of the tumor cells. In terms of mortality up to 16 or 17 days after inoculation, the response to challenge of day-old chicks with JMV-L did not appear to be influenced by the presence of antibody against MD related antigens but differed among strains of chickens. The estimated dose (LD50) that would kill 50% of a challenged population of Strain 4 was approximately 10,000 live JMV-L cells, while the corresponding estimates for strains 5 and K were approximately 600 cells and for strain S it was 50 cells. JMV-H and TLT were highly lethal to all four strains and all estimates of LD50 at 16 or 17 days post challenge were less than 10 cells. Resistance to JMV-L increased rapidly with age at challenge. Strains 4 and K were approaching complete resistance by 7 days of age and the susceptible strain S, when challenged at 14 days, was more resistant than day-old chicks of strains 4 and K. The chromosomal sex of the JMV-L, JMV-H and TLT tumor cells was female and this marker was used to confirm their transplantability.


Effect of polychlorinated biphenyls (Aroclor 1254) on liver gluconeogenic enzyme activities in embryonic and growing chickens. Fertile Nick Chick eggs were injected with 0.05, 0.1, 0.5 and 5 p.p.m. of PCB (Aroclor 1254) on day zero and were incubated until the 14th day of incubation. Nick Chick cockerels, one day old, were fed different levels of PCB in their diet for two weeks. The levels of PCB used were 0.1, 1, 5, 100 and 200 p.p.m. Key gluconeogenic enzyme activities were measured in the liver of both experimental groups. In the liver of chick embryos and growing chickens the decrease of the activity of all key gluconeogenic enzymes, with the exception of G6P-ase in embyronic liver, and FDP-ase in that of growing chickens, was estimated. The inhibited gluconeogenesis may account for the embryonic death in eggs contaminated with PCB's.


Immune response in progressor and regressor strains of chickens at specific intervals after a primary challenge with Rous sarcoma virus. A progressor line and a regressor line of chickens were compared for development of immune response, after an initial inoculation of Rous sarcoma virus in the wing-web. Quantitative measures of immunity were determined for the two contrasting lines of chickens at specific intervals after the initial challenge. It was found that subsequent challenges, with either Rous sarcoma virus or tumor homogenate, in the opposite wing-web were met by an early immune response from the regressor line. This early response was absent in the progressor line. However, both lines showed a similar immune response after a time interval of 15 days from the initial challenge. The genes for regression exert this early immune response which is the key to spontaneous regression of Rous sarcomas.


Comparison of tracheal histopathology and scanning electron microscopy of infectious laryngotracheitis. Samples of trachea were examined with light microscope, fluorescent microscope, and scanning electron microscopy (SEM). Negative controls and infectious laryngotracheitis (ILT) positive samples were compared. Histopathology illustrated lesions characteristic fo ILT. Lesions seen with SEM suggested surface changes including ciliary disruption. luminal debris, epithelial slough, crevices, hemorrhage and exudate. Fluorescence was noted in infected tracheas throughout both trials using fluorescent antibody (FA) technique.


Age-influence variations in the levels of cholesterol and ATPase activity in the testes of prepuberal chicks. Changes in the levels of total, free and esterified cholesterol and ATPase activity were measured in male prepuberal Nigerian fowl. Results showed that in all cases, the testicular levels of these parameters increased to peak levels and decreased thereafter. Total and free cholesterol levels were maximal at 12 weeks while the esterification of cholesterol was maintained from 14 weeks through to the 16th week. The results of ATPase activity corroborated these findings. The data suggest that peak androgen formation occurred in these chicks between 14th--16th weeks of age. As evidenced by the higher percentages of esterified cholesterol at all the ages studied, it is suggested that the Nigerian male fowl is an early-maturing specie.


The infectivity of fluid from regressing Rous sarcomas. Fluid was withdrawn from the site of regressing Rous sarcomas in chickens and inoculated into the wing-webs of untreated chickens from three strains of chickens with divergent degrees of resistance to Rous sarcomas. The transfer of fluid initiated progressive tumors in all three strains of chickens. The infectivity of the fluid was apparently due to tumor cells and not to virus.


Intestinal calcium binding protein levels in the one month old turkey poult. Intestinal calcium binding protein CaBP) was studied in one month old poults which had been fed a vitamin D3-deficient diet unsupplemented or with 399, 600, 900 or 1200 I.U. vitamin D3 added per kg. diet. CaBP levels increased with vitamin D3 supplementation up to 600 I.U. but showed no further increase with higher dietary vitamin D3. Poults supplemented with 600 to 1200 I.U. vitamin D3/kg. diet had an average of 1.03 mg. CaBP/g. of duodenal mucosa cells collected. The calcium binding activity of these samples was approximately 0.9% per mg. protein and was highly correlated with CaBP concentration (r=0.94).


Immunization of chickens against adenovirus infection. Chickens were vaccinated with monovalent and polyvalent adenovirus vaccines using three different serotypes, types 1, 2, and 3. No cross-protection was elicited by the heterologous serotypes but satisfactory protection from lesions and the shedding of challenge virus was induced by the homologous serotypes in the monovalent and polyvalent vaccines. The virus neutralizing (VN) titers were occasionally lower where the polyvalent vaccines were used when compared to those from chickens given the monovalent vaccines. This suggested possible interference though protection did exist against the appearance of lesions. Where chickens were vaccinated at an early age and in the presence of congenital homologous VN antibodies, protection against the effects of infection was satisfactory whereas the VN antibody titers were low or insignificant by the assay methods employed. The vaccine virus was not shed beyond 28 days after vaccination. No physical effects of vaccination were noted in any of the chickens during the post-vaccination period.


Effect of infectious bursal disease virus on the immunological responsiveness of the chicken. The immunological response of chickens infected at 28 days of age with infectious bursal disease virus (IBDV) was examined. Cellular immunity was studied by the graft-versus-host reaction and delayed type hypersensitivity skin reactions to tuberculin. Antibody responses to sheep red blood cells (S-RBC's) were tested by a direct hemagglutination test. Circulating levels of immunoglobulins (Ig's) IgM, IgG and IgA were monitored by the radial immunodiffusion test. The results of the graft-versus-host reaction and delayed type hypersensitivity reactions were similar for both the infected and control chicks. IBDV significantly reduced the primary antibody response to S-RBC's, but had no marked effect on the secondary antibody response. Chicks infected with IBDV had normal levels of serum IgM and IgG, but significantly lower levels of IgA when compared to uninfected control birds.


The enzymic preparation of diphosphoinositides. A procedure for the preparation of diphosphoinositides is described. Triphosphoinositides isolated from bovine brain are hydrolysed by the triphosphoinositide phosphatase (EC 3.1.3.36) from Crithidia fasciculata in the presence of MgC12 and cetyltrimethyl-ammonium bromide. The diphosphoinositides produced are not degraded further and can be recovered from the reaction mixture in greater than 80% yield. The product is chromatographically pure and has the same structure (1-phosphatidylinositol 4-phosphate) as naturally occurring diphosphoinositides.


Organization and expression of early genes of simian virus 40. The early region of simian virus 40 codes for at least two immunologically related polypeptides: large-T and small-t, with apparent molecular weights of 90,000-100,000 and 15,000-20,000, respectively. Because small-t shares methionine-containing tryptic peptides with large-T, the two polypeptides are probably coded, in part, by a common nucleotide sequence. To locate the coding sequences for large-T and small-t in the DNA, the production of these proteins was examined after infection of CV-1 cells with wild-type and deletion mutants of simian virus 40. We found that a deletion at the distal portion of the early region alters the structure of large-T but not of small-t; but deletions within the region between map coordinates 0.59 and 0.55 result in an alteration or absence of small-t and a normal large-T. These findings have been rationalized by a model that proposes the existence of two early mRNAs, one coding for large-T and the other for small-t. Both mRNAs span virtually the entire early region; but the mRNA coding for large-T lacks the nucleotide sequence between map coordinates 0.59 and 0.54. We suggest that small-t is translated from the larger of the two mRNAs, beginning at or near its 5' end and terminating at a termination codon at about map coordinate 0.54. Larger-T, on the other hand, is translated from the shorter mRNA, beginning at the same initiator codon, and, because of the deletion of the terminator codon at 0.54, translation proceeds to the terminator codon at or near map position 0.18.


Spin labeling of a cysteine residue of the Escherichia coli outer membrane lipoprotein in its membrane environment. A method was developed to attach a spin label to a specific site on the structural lipoprotein of the Escherichia coli outer membrane in situ. This method takes advantage of the fact that the outer membrane of wild-type E. coli contains few residues reactive towards sulfhydryl reagents. A mutant E. coli strain has been isolated [Suzuki, H., Nishimura, Y., Iketani, H., Campisi, J., Hirashima, A., Inouye, M. & Hirota, Y. (1976) J. Bacteriol. 127, 1494-1501] in which the second position from the carboxy terminus of the lipoprotein is changed from arginine into a cysteine residue. The membrane fraction of this mutant was treated with N-(1-oxyl-2,2,5,5-tetramethylpyrrolidinyl)maleimide in the presence of EDTA and 2-mercaptoethanol. Spin label was found to be preferentially incorporated into the lipoprotein. The spectrum of the spin-labeled membrane shows two components, both arising from spin label at the same site near the carboxy terminus. The strongly immobilized component has a maximum hyperfine splitting value of 53 G, and the weakly immobilized component, 37 G. A fraction of the lipoprotein is covalently bound to the peptidoglycan layer through its carboxy-terminal lysine; the spectrum of the isolated bound form of the lipoprotein was identical to that of the free form. When the matrix protein, the other major outer membrane protein, was removed by mutation, the spectrum of the lipoprotein was altered, suggesting that these two proteins are closely associated.


Similarity of nucleotide sequences around the origin of DNA replication in mouse polyoma virus and simian virus 40. The nucleotide sequence around the origin of replication in DNA of mouse polyoma virus was determined by 32P labeling of the 3' terminus of the Hap II-5/Alu I-1 DNA fragment, with the use of DNA polymerase. The result coincided with our previous report on the 32P labeling, with the use of polynucleotide kinase, of the 5' terminus of the Hap II-5/Hha I-1 DNA fragment, which corresponds to the large part of the present fragment, Hap II-5/Alu I-1. A symmetrical (A+T)-rich region containing a five-A stretch (or a five-T stretch) was flanked by two small regions with a 2-fold rotational axis of symmetry. On comparison of the sequence near the replication origin of polyoma DNA with that in the corresponding region of simian virus 40 DNA, which was included in the EcoRI-G fragment sequenced by Weissman's group (Subramanian K.N., Dahr, R. & Weissman, S. M. (1977) J. Biol. Chem. 252, 355--367), a considerable similarity was detected. Several possible common sequences for important biological activities such as the starting of DNA replication and RNA synthesis were suggested.


Isotopic (18O) shift in 31P nuclear magnetic resonance applied to a study of enzyme-catalyzed phosphate--phosphate exchange and phosphate (oxygen)--water exchange reactions. An isotopic shift of the (31)P nuclear magnetic resonance due to (18)O bonded to phosphorus of 0.0206 ppm has been observed in inorganic orthophosphate and adenine nucleotides. Thus, the separation between the resonances of (31)P(18)O(4) and (31)P(16)O(4) at 145.7 MHz is 12 Hz and, in a randomized sample containing approximately 50% (18)O, all five (16)O-(18)O species are resolved and separated from each other by 3 Hz. Not only does this yield the (18)O/(16)O ratio of the phosphate but, more important, the (18)O-labeled phosphate in effect can serve as a double label in following phosphate reactions, for oxygen in all cases and for phosphorus, provided the oxygen does not exchange with solvent water. Thus, it becomes possible to follow labeled phosphorus or labeled oxygen continuously as reactions proceed. Rate studies involving (i) phosphorus and (ii) oxygen are illustrated by continuous monitoring of the exchange reactions between (i) the beta phosphate of ADP and inorganic phosphate catalyzed by polynucleotide phosphorylase and (ii) inorganic orthophosphate and water catalyzed by yeast inorganic pyrophosphatase. In the ADP-P(i) exchange, the P(i) ((18)O(4)) yielded an alpha P((16)O(3) (18)O) and a beta P((18)O(4)), proving that bond cleavage occurs between the alpha P and the alpha-beta bridge oxygen. Among the many additional potential uses of this labeling technique and its spectroscopic observation are: (i) different labeling of each phosphate group of ATP, (ii) to follow rate of transfer of (18)O from a nonphosphate compound such as a carboxylic acid to a phosphate compound, and (iii) to follow the rate of scrambling (for example, of the beta-gamma bridge oxygen of ATP to nonbridge beta P positions) and simultaneously the rate of exchange of the gamma P nonbridge oxygens with solvent water in various ATPase reactions.


Specific regulation by steroid hormones of the amount of type I cyclic AMP-dependent protein kinase holoenzyme. The total amounts of type I and type II cytoplasmic cyclic AMP-dependent protein kinase activities were measured in various tissues of intact rats and rats subjected to castration, hypophysectomy, or adrenalectomy. After castration, the total amount of type I activity decreased rapidly in classifically steroid-responsive tissues such as the ventral prostate and levator ani muscle and less rapidly in the liver. After hypophysectomy and adrenalectomy, type I activity in the liver decreased to the same extent as after castration. Type I activity could be maintained in the ventral prostate and levator ani muscle at control levels by the daily injection of dihydrotestosterone. Furthermore, after post-castration regression of the prostate for 3 days, three daily subcutaneous injections of dihydrotestosterone resulted in a complete restoration of type I activity toe the intact level. The amount of type II activity was not altered by any of the experimental ablations. This study provides evidence linking steroid action to the ability of steroid-responsive tissues to maintain a substantial activity of type I cyclic AMP-dependent protein kinase.


Uracil incorporation: a source of pulse-labeled DNA fragments in the replication of the Escherichia coli chromosome. Uracil is incorporated into newly synthesized DNA by mutants of Escherichia coli with reduced levels of dUTPase (dUTP nucleotidohydrolase; EC 3.6.1.23). Excision-repair of the incorporated uracil results in the generation of labeled DNA fragments that appear after brief pulses with [(3)H]thymidine [Tye, B-K., Nyman, P.-D., Lehman, I. R., Hochhauser, S. & Weiss, B. (1977) Proc. Natl. Acad. Sci. USA 74, 154-157]. Uracil is also incorporated into the newly synthesized DNA of strains of E. coli that contain normal levels of dUTPase. DNA fragments generated by the postreplication excision-repair of uracil may therefore contribute to the pool of nascent DNA (Okazaki) fragments that normally appear in wild-type strains. Discontinuous DNA replication has been examined in the absence of uracil excision by comparing Okazaki fragments in strains that are defective in DNA polymerase I (polA(-)) and polA(-) strains that are also defective in uracil N-glycosidase, an enzyme required for the excision-repair of uracil in DNA (polA(-)ung(-)). Little or no difference was detected in the level of Okazaki fragments in the polA(-) strain as compared with the polA(-)ung(-) strain. Thus, the uracil-induced cleavage of DNA cannot be the sole mechanism for the generation of Okazaki fragments. Mutants that are defective both in dUTPase and in uracil N-glycosidase incorporate uracil into their DNA with a high frequency (up to 1 per 100 nucleotides). These uracil residues, once incorporated, persist in the DNA without an adverse affect on the growth of the cells.


DNA intermediates at the Escherichia coli replication fork: effect of dUTP. We have directly tested the hypothesis that elevated levels of dUTP cause the formation of small DNA fragments at the replication fork of Escherichia coli. Addition of increasing levels of dUTP to lysates on cellophane discs results in an increasing number of strand scissions in the newly replicated DNA. Lysates of strains defective in dUTPase produce many more scissions at the same level of dUTP. The size distribution of Okazaki pieces obtained in vivo can be reconstituted in vitro on cellophane discs if appropriate levels of dUTP are present. Although uracil excision leads to the apparent production of Okazaki pieces from both daughter strands, DNA synthesis is actually asymmetric under these conditions. De novo chain initiation events occur on only one strand. It is suggested that asymmetry of synthesis in vivo may be masked by uracil excision and other postreplication processing mechanisms.


In vivo phosphorylation of a synthetic peptide substrate of cyclic AMP-dependent protein kinase. A model synthetic peptide substrate of the cyclic AMP-dependent protein kinase (ATP:protein phosphotransferase; EC 2.7.1.37), Leu-Arg-Arg-Ala-Ser-Leu-Gly, closely resembling the local phosphorylation site sequence in porcine hepatic pyruvate kinase, was shown to be phosphorylated in vivo after microinjection into Xenopus oocytes. This result demonstrates that the microinjection technique, utilizing a synthetic peptide substrate, or possibly a synthetic substrate analog inhibitor [Kemp, B. E., Benjamini, E. & Krebs, E. G. (1976) Proc. Natl. Acad. Sci. USA 73, 1038--1042], can be used to study protein phosphorylation-dephosphorylation reactions in living oocytes. This follows, since it is clear that the injected peptide was accessible to the cellular compartment containing the protein kinase.


Evidence for the presence of two nonidentical subunits in NAD-dependent isocitrate dehydrogenase of pig heart. The NAD-dependent isocitrate dehydrogenase [threo-D(S)-isocitrate:NAD(+) oxidoreductase (decarboxylating); EC 1.1.1.41] from pig heart is a multisubunit enzyme with a molecular weight of approximately 340,000. Electrophoresis of the enzyme in 10% polyacrylamide gels containing sodium dodecyl sulfate reveals two discrete bands with molecular weights of 41,000 and 39,000. The two bands exhibit approximately equal intensity when stained with Coomassie Blue, Amido Black, and Bromophenol Blue, suggesting that these polypeptide chains are present in equimolar quantities in the native enzyme. The same two-band pattern is observed when the sulfhydryl groups of the enzyme are blocked by alkylation with iodoacetate prior to electrophoresis, indicating that sulfhydryl oxidation is not responsible for the observed heterogeneity. Each of the subunits appears as a single band when eluted from the gel and again subjected to electrophoresis under the same conditions. Isocitrate dehydrogenase contains a total of 41 lysine and arginine residues per average subunit of 40,000 daltons. The observation of approximately 80 peptides upon paper chromatography and high voltage electrophoresis of tryptic digests of the enzyme is consistent with the existence of two distinct polypeptide chains. Dansylation yields two NH(2)-terminal amino acid derivatives: dansyl-phenylalanine and dansyl-alanine. It is concluded that the NAD-specific isocitrate dehydrogenase is composed of equal numbers of two nonidentical subunits.


Relationship between dibutyryl cyclic AMP and microtubule organization in contracting heart muscle cells. Rhythmic and synchronous contractions of interconnecting myocyte cultures prepared from fetal rat hearts are arrested upon the addition of 0.2--2.0 mM N6, O2'-dibuturyladenosine 3':5'-cyclic monophosphate (Bt2cAMP). The contractions arrested by Bt2cAMP are restored either by diluting the Bt2cAMP from the media or by adding colchicine. While colchicine restores Bt2cAMP-arrested myocyte contractions at concentrations as low as 1.0 micron, the inactive isomer lumicolchicine shows no effect. Morphologically, Bt2cAMP treatment of myocyte cultures results in the appearance of numerous elongated cellular processes not present in control cultures. Ultrastructural examination indicates that in Bt2cAMP-treated cells the intracellular distribution of microtubules is altered such that these organelles appear to accumulate in parallel arrays. In cells not treated with Bt2cAMP, the microtubules appear randomly oriented, while in cells treated with only colchicine, intact microtubules are not observed. The relationship between microtubules an heart cell contraction is discussed.


Carcinogens enhance survival of UV-irradiated simian virus 40 in treated monkey kidney cells: induction of a recovery pathway? Treatment of monkey kidney cells with low doses of carcinogen enhances the survival of UV-irradiated simian virus 40 (SV40). This is true for compounds with UV-like effects (metabolites of aflatoxin B1, N-acetoxyacetylaminofluorene) and compounds with x-ray-like effects (methyl methanesulfonate, ethyl methanesulfonate). This phenomenon resembles the UV-reactivation of viruses in eukaryotic cells. The carcinogen-induced enhancement of the survival of UV-irradiated SV40 is correlated with the inhibition of host-cell DNA synthesis, suggesting that the inhibition is an inducing agent. An enhancement of UV-irradiated SV40 survival is also obtained in cells treated with hydroxyurea or cycloheximide for long enough that there is still inhibition of host DNA synthesis during the early stage of SV40 infection. We hypothesize that treatment of host cells with carcinogens induces a new recovery pathway that facilitates the replication of damaged DNA, bypassing the lesions and resulting in the enhanced survival of UV-irradiated SV40. This inducible process might represent the expression of "SOS repair" functions in eukaryotic cells analogous to the previously demonstrated induction of SOS repair in bacteria after UV or carcinogen treatment.


Role of contact inhibition in the regulation of receptor-mediated uptake of low density lipoprotein in cultured vascular endothelial cells. Bovine vascular endothelial cells during logarithmic growth bind, internalize, and degrade low density lipoprotein (LDL) via a receptor-mediated pathway. However, contact-inhibited (confluent) monolayers bind but do not internalize LDL. This is in contrast to aortic smooth muscle cells or endothelial cells that have lost the property of contact inhibition. These cells internalize and degrade LDL at both high and low cell densities. The LDL receptors of smooth muscle and sparse endothelial cells down-regulate in response to LDL. In contrast, normal endothelial cells at confluency show little response. When contact inhibition in endothelial monolayers was locally released by wounding, and LDL was present, only cells released from contact inhibition accumulated LDL cholesterol. In smooth muscle cells under the same conditions, the entire culture interiorized lipid. It thus appears that in endothelial cells, unlike smooth muscle cells, contact inhibition is the major factor regulating cellular uptake of LDL cholesteryl ester. Reversal of contact inhibition by wounding provides a mechanism by which the endothelium could be the primary initiator of the atherosclerotic plaque.


Multiple forms of sarc gene proteins from Rous sarcoma virus RNA. In a previous study we were able to identify two proteins of 25,000 and 18,000 daltons that were made from RNA of transforming virions of Rous sarcoma virus (RSV) and that were missing from the translation products of a transformation-defective deletion mutant of RSV. In the present study we have separated RSV virion RNA on sucrose gradients and have determined that the two putative sarc gene products are synthesized as doublets from an mRNA of approximately 18 S. There also appear to be several other sizes of virion mRNA that direct the synthesis of other viral proteins. These data are discussed in terms of the structure of the RSV genome. In addition to the 25,000- and 18,000-dalton doublets, there also is a 60,000-dalton protein whose synthesis is directed by 18S viral RNA from transforming virion of RSV. Peptide mapping has shown that the 60,000- and 25,000-dalton doublet are structurally related. In addition, the use of two-dimensional gel electrophoresis has allowed us to resolve both bands of the 25,000-dalton doublet into several differently charged species.


Structural flexibility of isozyme variants: genetic variants in Drosophila disguised by cofactor and subunit binding. Wild populations of Drosophila mojavensis exhibit considerable conformational variation in the NAD+-free form of alcohol dehydrogenase (alcohol:NAD+ oxidoreductase; EC 1.1.1.1). The variation appears genetic, as it does not occur within an inbred strain. The NAD+-bound form of alcohol dehydrogenase, present in the same individuals, does not exhibit the variation, suggesting that the binding of NAD+ acts to stabilize conformation. Such cofactor binding to enzymes may thus conceal considerable variation. A similar effect is suggested for binding of esterase subunits.


Analysis of human tumors and human malignant cell lines for BK virus-specific DNA sequences. Most humans in the United States have been infected with BK virus (BKV), a human papovavirus. Because BKV has oncogenic properties, we have investigated whether it may be a cause of human cancer. Basic principles of tumor virology imply that BKV-induced tumors should contain BKV DNA sequences. Therefore, we assayed (by molecular hybridization) DNA from human tumors and malignant cell lines for BKV DNA, using BKV [(32)P]DNA as probe. The BKV [(32)P]DNA was labeled in vitro (nick translation) to specific activities of 1 to 2 x 10(8) cpm/mug. The BKV DNA used to prepare our probes had the properties expected of authentic BKV genomes, including density of superhelical DNA, sedimentation velocity in alkaline and neutral sucrose gradients, production of one fragment by endonuclease EcoRI cleavage and four fragments by endonuclease Hin II + III cleavage and reassociation properties. From these studies we conclude that our BKV probes hybridized well, and represented bona fide BKV DNA. Using three different BKV [(32)P]DNA probes, i.e., from three distinct plaque isolates, we have analyzed DNA from BKV-transformed cells, normal human tissues, and a large number of human tumors. All human DNAs (cell lines, normal tissues, tumors) hybridized 5% with BKV DNA. Hybridization analysis of BKV-transformed hamster cell DNA indicated 5-6 copies of at least 88% of the BKV genome per cell. No BKV DNA sequences were detected (above the normal 5% hybridization to all human DNAs) in the following normal human tissues: 10 kidney (BKV is usually isolated from urine), 3 spleen, 13 lung, 23 colon, 2 rectum, 1 ileum, and 1 skin. No BKV-specific DNA was found in 166 tumors, including 5 carcinomas (Ca) of stomach, 3 Ca small intestine, 26 Ca colon, 9 Ca rectum, 31 Ca lung, 9 adenocarcinomas and 5 oat cell carcinomas of lung, 17 melanomas, 5 Ca prostate, 4 Ca bladder, 6 Wilms tumors, 4 hypernephromas, 15 Ca kidney, 7 brain tumors, 5 Hodgkin lymphomas, 10 lymphomas (immunosuppressed patients have a high incidence of lymphomas), 2 reticulum cell sarcomas (spleen), and 3 skin tumors. We have also analyzed 7 human malignant cell lines (melanoma, lung, rhabdomyosarcoma, and glioblastomas), including several clones of a lung melanoma line; no BKV DNA sequences were detected. Because our probes could detect one copy of BKV DNA if only 10% of the cells were tumor cells, our results are very strong evidence that the tumors we analyzed did not have a BKV etiology. The tumors we tested represent about 50% of all cancers in the United States; there is no evidence that BKV is involved in the etiology of these types of tumors.


Evidence for X-linkage of human phosphoribosylpyrophosphate synthetase. The mode of genetic transmission of human phosphoribosylpyrophosphate synthetase (ribosephosphate pyrophosphokinase; ATP:D-ribose-5-phosphate pyrophosphotransferase; EC 2.7.6.1) was studied in fibroblasts cultured from members of a family with a structurally and electrophoretically altered phosphoribosylpyrophosphate synthetase that has increased activity per enzyme molecule. Enzyme activity in fibroblast lysates from the daughter of an affected male patient was intermediate to the activities in lysates from her father (and her affected paternal uncle) and from her mother and other normal individuals. Two bands of enzyme activity corresponding to normal and mutant phosphoribosylpyrophosphate synthetases were found in fibroblast lysates from the daughter after cellulose acetate strip electrophoresis. In contrast, only mutant enzyme was detectable in lysates derived from the male patients. Fibroblasts cloned from the daughter contained two phenotypically distinct (normal and mutant) populations of cells with respect to phosphoribosylpyrophosphate synthetase activity and electrophoretic mobility. These studies support assignment of the structural gene for human phosphoribosylpyrophosphate synthetase to the X-chromosome. No evidence for the presence of the normal enzyme was found in erythrocyte or lymphocyte lysates or in partially purified erythrocyte enzyme preparations from the heterozygous daughter, suggesting either nonrandom X-chromosome inactivation in precursors of these cells or selection against hematopoietic cells bearing the normal enzyme after random X-chromosome inactivation.


Characterization and mapping of RNase T1-resistant oligonucleotides derived from the genomes of Akv and MCF murine leukemia viruses. T1 RNA fingerprints of the genomes of Akv-1 and Akv-2 C-type viruses are indistinguishable and oligonucleotide maps of these viruses are probably the same. Akv-1 and -2 share 55--75% of their large T1-resistant oligonucleotides with four MCF viruses isolated from AKR mice or from NIH Swiss mice that inherit either the Akv-1 or Akv-2 virus-inducing locus of AKR. The majority of Akv oligonucleotides missing from T1 fingerprints of MCFs and the majority of oligonucleotides unique to MCF viruses are clustered and  lie at corresponding positions in the 3' half of the oligonucleotide maps of Akv and MCF viruses. The RNA sequences present in different MCF isolates but not present in Akv-viruses are related. These results are consistent with a recombinational origin of MCF viruses, as proposed by Hartley and Rowe and their collaborators.


Transcription of a defective polyoma virus genome. The circular genome of the cloned defective polyoma virus D-50 consists of tandemly repeated copies of the DNA sequence between 67 and 84 units on the wild-type polyoma virus DNA map. Each repeated copy thus contains the origin of viral DNA replication, which is located at about 71 map units. Viral RNA was synthesized in vitro using viral transcription complexes extracted late (30 hr) after infection from mouse cells co-infected with D-50 and helper wild-type virus. Both wild-type and D-50 DNA molecules were active as templates for in vitro transcription. Approximately 84% of the RNA transcribed in vitro from wild-type DNA was complementary to the L DNA strand. This is normal for wild-type transcription late after infection. By contrast, at least 90% of the RNA transcribed from D-50 DNA molecules was complementary to the E DNA strand. After normalization of the data to account for the observed molar ratio of D-50 DNA repeated sequences to unit length wild-type DNA, we estimate that transcription of the E DNA strand of each D-50 repeated unit is about 1.4 times as efficient as transcription of the wild-type E DNA strand. Transcription of the D-50 L DNA strand, however, is only 0.03 times as efficient as transcription of the wild-type L DNA strand. The implications of these results concerning the nature and location of promoter sequences in polyoma DNA are discussed.


Tumor antigen(s) in cell productively infected by wild-type polyoma virus and mutant NG-18. When isolated by means of an anti-polyoma tumor (T) antiserum, the major product from mouse cells productively infected by wild-type polyoma virus is a polypeptide of 100,000 apparent molecular weight as judged by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. In cells infected by NG-18, an hr-t mutant carrying a deletion of about 150 base pairs in the early region of the viral DNA, a T antigen species appears that comigrates with that of the wild-type virus. Comparisons of peptides after partial proteolysis reveal no differences between mutant and wild-type products. Both wild-type and mutant 100,000 products can be labeled in vivo with [(32)P]orthophosphate. An independent and more reliable estimate of the molecular weight of this protein using guanidine/Sepharose chromatography yields a value of 81,000 for both mutant and wild-type species. The apparent identity of wild-type and mutant products indicates that the deletion in NG-18 lies outside of the region encoding this major T antigen species. Immunoprecipitates from wild-type infected cells shows four bands in addition to the "100,000" band; these have apparent molecular weights of 63,000, 56,000, 36,000, and 22,000 by sodium dodecyl sulfate/polyacrylamide gel electrophoresis; the 56,000 and 36,000 species are phosphorylated. All four of these lower molecular weight bands are absent or drastically reduced in the immunoprecipitates from NG-18-infected cells.


Gonadotropin binding and stimulation of steroidogenesis in Leydig tumor cells. Testicular tumors are generally characterized by a loss of responsiveness to gonadotropins. The M5480 Leydig cell tumor is unusual, if not unique, in that it responds to human choriogonadotropin and to lutropin via increased steroidogenesis. This report describes the identification of two variants of the original M5480 tumor that have altered steroid output both in the basal state and in response to human choriogonadotropin. One of the tumors produces mainly progesterone, which is stimulated by the choriogonadotropin; the other tumor produces about equal amounts of progesterone and testosterone, and the secretion of both is stimulated by the choriogonadotropin. The dissociation constant describing the interaction between Leydig tumor cells and (125)I-labeled human choriogonadotropin is between 3 and 5x10(-11) M. This agrees with values reported for normal Leydig cells, although the tumor cells appear to have fewer receptors. The differences noted in the two tumors and normal Leydig cells may have arisen from alterations in gene regulation, or in mutations, involving one or more enzymes in the pathway in which progesterone is converted to testosterone. Under the experimental conditions used, all the tumors studied (seven generations) responded to the choriogonadotropin both in binding and in the resultant stimulation of steroidogenesis. This property, together with the characteristic that a homogeneous cell population can be obtained without enzymatic treatment, should qualify the M5480 Leydig cell tumor(s) as a model system for further studies on the mechanism of action of gonadotropin, on hormone receptors, and on hormonally responsive tumors.


Nutritional agents which affect metabolic zinc status. As the number of factors which affect zinc absorption and homeostasis increase, it becomes more apparent that the relative zinc status of the population cannot be estimated from the analyzed zinc content of foods. The environment of the small intestine at the time of absorption can only be estimated from the diet fed. Even then since both endogenous and exogenous factors determine the zinc absorbed, the precision of these estimations should allow for large individual variation.


Central catecholamine and peripheral noradrenaline depletion: effects on one-way trace-conditioning. The present study evaluated the differential contributions of the central and peripheral catecholaminergic systems in aversive learning using trace-conditioning procedure. Twenty-seven male Wistar rats were randomly assigned to one of the three groups: central drug and peripheral saline (CDPS) injections, central saline and peripheral drug (CSPD) injections, and central saline and peripheral saline (CSPS) injections. Results showed that: (1) the rate of acquisition and the overall avoidance responses for the drug treated groups was significantly poorer than the control group; (2) there was no significant difference in performance between the two drug treated groups; and, (3) neither central nor peripheral catecholamine depletion had a significant effect on the secretion of plasma corticosterone. The findings failed to confirm the hypotheses (1) that central CA depletion is more detrimental than peripheral NA depletion, (2) that plasma 11-OHCS plays a mediating role in the acquisition of avoidance and escape responses in rats depleted of catecholaminergic systems.


On the mechanism of renin release by restraint stress in rats. Restraint causes an increase in plasma renin activity (PRA) which is not affected by pretreatment with dl-propranolo (1 mg/kg IP) or sotalol (15 mg/kg IP). These doses of beta-adrenergic blocking agents are effective in suppressing the stimulation of PRA by isoproterenol. Large doses of dl-propranolol (10 mg/kg IP) and d-propranolol (5 mg/kg IP) attenuate the restraint-induced PRA increase. Adrenal demedullectomy does not affect the PRA response to restraint. Renal denervation blunts the PRA rise due to restraint, but not to direct stimulation by the beta-adrenergic agonist, isoproterenol. It is concluded that the increase in PRA during restraint stress in rats is not solely dependent on an intact renal sympathetic innervation. A significant portion of this stress-induced PRA increase appears to involve a non-adrenergic mechanism.


Morphine-induced tail erection: site of action. Acute ablation techniques were used to localize morphine-induced tail erection (MITE) within the central nervous system of mice. Morphine produced no elevation of tails in mice whose spinal cord had been transected at the lower thoracic or lumbar levels. Decortication and high-level precollicular decerebration did not prevent MITE while morphine caused no tail response at all in low-level inferior collicular decerebrate mice. Lesions in various portions of the mesencephalon revealed that the degree of MITE was closely related to the size of the lesions of the central gray matter. The larger the lesion, the smaller the degree of tail elevation. MITE could not be elicited in mice when the mesencephalic central gray matter had been completely destroyed. Results indicate that morphine acts on the mesencephalic central gray matter producing tail erection and the pathway responsible for the reaction descends from the midbrain downward to the spinal cord.


Cortical EEG power spectra associated with sleep-awake behavior in the rat. Power spectral analyses were used to study cortical EEG activities during sleep-awake behavior in the rat. EEG spectra, both long-time and sequential short-time, derived from EEG during the states of wakefulness, sleep, and REM sleep were qualitatively and quantitatively different. The degree of inter- and intrasubject variability between these spectra was minimal. This experimental model with the rat should allow quantitative delineation of cortical EEG changes produced by psychotropic drugs.


Baseline sleep-wake patterns in the pointer dog. The 24-hr electrographic patterns (EEG, EOG, EMG) of six normal pointer dogs were recorded in a laboratory setting. Two states of sleep (slow-wave and rapid eye movement) and wakefulness (alert and drowsy) were identified. The total recording period comprised 44% of alert wakefulness, 21% of the drowsy state, while slow-wave sleep occupied 23% and REM sleep 12% of the time. The mean length of a REM sleep episode averaged 6 min and the mean REM sleep cycle was 20 min. The mean polycyclic sleep-wake cycle was 83 min. Sleep episodes averaged 45 min and the mean waking episode was 38 min. There was an average of two REM sleep episodes per sleep-wake cycle. The dog has a propensity to sleep over a 16-hr interval from 1300 to 0500 but the most sleep occurred between 2100 and 0400 hr during darkness.


Sleep induced by intestinal stimulation in cats. The influence of afferent impulses of intestinal origin on the sleep stages was studied in fed and starved cats. Low-frequency electrical stimulation of the mucosal surface in a small intestinal fistula reduced the latency of sleep onset. The number of slow wave sleep episodes decreased, but their mean duration increased during stimulation. Conversely, the number of paradoxical sleep episodes increased, but their mean duration was not significantly modified by the intestinal stimulation. The role of viscerosensory events in the control of sleep is discussed in relationship to these results.


Two preliminary studies on sleep and psychotherapy. Two preliminary studies were conducted to assess the effects of an intensive outpatient psychotherapy, Feeling Therapy, on sleep. This therapy was chosen because of its demonstrated ability to affect its patients' dreams. In the first study a newly entering female patient was recorded across the first three weeks of intensive daily therapy. In contrast to two control subjects recorded across a similar time period, she demonstrated low REM times and short REM latencies on the average, and considerably greater variability in nearly every parameter. In the second study, two patients were recorded across three days (the middle of which was the day of a therapy session) first when new in therapy and then again after two and one-half years of therapy. It was found that when new in therapy both subjects spent nights of significantly altered sleep the day of the therapy session. One subject showed no REM sleep whatsoever while the other showed a 10 min REM latency and low REM time. The significance of these findings and the direction of future research is discussed.


Failure of interocular transfer in the pigeon (Columba livia). Pigeons were trained on a modified jumping stand to discriminate a circle from a triangle using only one eye. Testing with the naive eye revealed no evidence of interocular transfer. This failure of transfer was also observed under more stringent testing using reversal learning. The results are discussed in terms of a selective attention model for panoramic vision.


Ulcer reduction by non-nutritive bulk in pylorus ligated rats. The effects of solid non-nutritive diet on pylorus-ligation ulcers were tested. An experimental group of 16 rats received a bulky mixture of silica and methylcellulose during 48 hr prior to ligation. Control rats were food deprived for the same period of time. A multivariate analysis of variance was applied to nine measures of the study. It was found that rumenal ulceration and total acidity were lower in experimental animals than in controls. This finding confirms a previous observation indicating that the physical property of diet has antiulcerogenic effects.


[Phasic activity in rats].

[Phasic activity in rats]. At the central level, in the rat, phasic activity has been recorded during paradoxical sleep and in acute conditions after injection of reserpine or parachlorophenylalanine. At the external level, during paradoxical sleep, the extraocular muscles lateral rectus, superior rectus and superior oblique are activated in both plastic and tonic manners. The muscles of the whiskers are also activated; these muscular activations are more often than not synchronous with the eye movements (80%). The time distribution of these ocular movements is homogenous. Reserpine induces phasic muscular activations of the extraocular muscles.


Ulnar artery thrombosis.

Ulnar artery thrombosis. Forty-six patients with ulnar artery thrombosis were evaluated. A comparison of the results was made between those patients who were treated by sympathectomy and those in whom we attempted to reestablish the blood flow. A reverse vein graft was often used in the latter group. Based on the results of this study, we believe the preferred method of treatment for ulnar artery thrombosis is an attempt at the reestablishment of blood flow.


A psychophysiological investigation of the long-term effects of clozapine upon sleep patterns of normal young adults. A 25-night single-blind cross over design was employed to determine the long-term effects of clozapine on the sleep patterns of six normal young adults. Subjects received 12.50 mg placebo on the first and last five nights, whereas on the intermediate 15 nights 12.5 mg clozapine was administered. The subjects slept in the laboratory on the third and fourth nights to obtain baseline recordings, and on the eight, twelfth, sixteenth, and twentieth nights to determine the effects of clozapine on sleep variables. Recordings on nights 21 and 25 were used to assess withdrawal effects. Percentage stage 1 sleep and indices of body movements during sleep were significantly reduced, suggesting that clozapine may have sleep-inducing properties. There were no significant rebound of stage REM sleep during drug withdrawal despite a small but significant reduction in stage REM during drug administration. Numerous side effects, indicative of sleepiness, were reported on the mornings following drug administration, and there was evidence of a rapid tolerance to clozapine. These findings may limit the efficacy of clozapine as an hypnotic agent over an extended period of time. Further research on insomniac subjects is therefore indicated.


Screening of a family for chemodectoma.

Screening of a family for chemodectoma. 59 relatives with a family history of hereditary chemodectoma have been successfully screened by means of radionuclide scintiangiography. The findings showed a close correlation with selective roentgenangiography. 3 asymptomatic patients with chemodectoma were revealed by these methods without causing discomfort to the group involved.


Diagnosis of juvenile angiofibroma by computed tomography. Computed tomography (CT) accurately localized juvenile angiofibromata in 3 patients. The expanded pterygopalatine fossa and canal were visualized by CT in all three cases. Because of the hemorrhagic tendency of these tumors, a noninvasive modality such as CT is especially valuable in planning therapy.


99mTc-pyrophosphate imaging in acute pericarditis: a clinical and experimental study. Fifteen patients with clinical and electrocardiographic features of acute pericarditis underwent myocardial scintigraphy using 99mTc-pyrophosphate. All had normal images. In 5 additional patients with acute pericarditis and evidence of ischemic heart disease, 99mTc-pyrophosphate images showed focal abnormalities in 2 patients and equivocal findings in 2. Serial myocardial radionuclide images were obtained 2 to 18 days after induction of pericarditis in 8 dogs; all images were normal. No stainable tissue calcium was demonstrated histochemically in the pericardium or myocardium of these dogs. Our results suggest that 99mTc-PYP myocardial radionuclide images are normal in acute pericarditis in the absence of ischemic heart disease.


Thallium-201 distribution in the thyroid: relationship to thyroidal trapping function. Experiments were undertaken to determine the behavior of thallium-201 in the thyroid in relation to the iodide-trapping mechanism in rats. Iodide trapping was compared to thyroidal thallium-201 trapping 30 minutes after intravenous injection. Thallium-201 uptake followed iodide uptake in control rats. TSH- and T3-treated rats but did not decrease in the same manner as iodide concentration in perchlorate-treated rats. These studies suggest that thallium-201 distribution in the thyroid may reflect the cationic side of the iodide pump or possible thyroidal blood flow.


Dibutryl cyclic adenosine monophosphate: effect on radiosensitivity of tumors and normal tissues in mice. The effect of dibutryl cyclic adenosine monophosphate (db-cAMP, 30 mg/kg), injected into mice before irradiation, on the radiation sensitivity of various tissues was investigated. Survival of proliferating hair follicles, small gut stem cells, fibrosarcoma (FSa) micrometastases in the lungs, and two different mammary carcinomas, as assayed by the TCD50 method, were studied. Intraperitoneal injection of db-cAMP before irradiation resulted in higher survival of hair follicles irradiated on the third day after plucking. Stem cells in the small gut showed increased survival if irradiated 4 hours after injection. There was no effect on the survival of FSa micrometastases by preinjection and the TCD50/120 days of the mammary carcinomas was not altered.


The isolation of adenoviruses from goats affected with peste des petits ruminants in Nigeria. Mucosal scrapings from the large intestine of two goats that had died from peste des petits ruminants (PPR) in separate outbreaks in Nigeria were examined for viruses. A mixed viral infection of PPR virus (morbillivirus) and adenovirus was confirmed in both goats. The adenoviruses did not conform to any of the ovine and bovine serotypes recognised; the two isolates were considered different serotypes. It is concluded that, although the role of adenoviruses in the epizootiology of PPR in Nigeria is difficult to appraise, they are probably commensals. This is believed to be the first report of the isolation of adenoviruses from goats.


Isolation of avian infectious bronchitis virus from experimentally infected chickens. Virus was recovered from the faeces of chickens infected at three or four weeks of age for more than 20 weeks after infection with commercial vaccines or with the T strain of avian infectious bronchitis virus (IBV). Virus was not recovered from the trachea, liver, spleen, bursa or kidneys of T strain infected birds longer than 29 days after infection at which point IBV was recovered from the bursa of a single infected bird. In a subsequent experiment IBV was recovered from the caecal lymph nodes and faeces of one of five birds 14 weeks after infection with a commercial vaccine but no virus was isolated from the trachea, kidneys, duodenum, bursa, ovaries or testes of any of the five birds at this time.


Subclinical pneumonia associated with an experimental adenovirus infection in the domestic fowl and the effect of concurrent infectious laryngotracheitis virus. A CELO-type adenovirus (AV) isolated from fowls with respiratory disease was inoculated experimentally into the tracheas of young birds. No symptoms referable to respiratory infection were evident. Post mortem examination between days 2 and 5 after inoculation revealed pneumonia involving up to 30 per cent of the surface of the lungs. Histologically, a focal to diffuse interstitial lymphocytic infiltration and bronchiolar degeneration were present. Concurrent infections with a mild strain of infectious laryngotracheitis virus (ILT) failed to enhance the pathogenicity of either the AV or ILT infections.


Lack of maedi viral related RNA in pulmonary carcinoma of sheep (jaagsiekte). Molecular hybridisation with radioactively labelled DNA complementary to the RNA of the maedi virus was used to probe for homologous RNA in the polysome fraction of pulmonary carcinomas (jaagiekte) of Awassi sheep. No sequence homology was detected, which suggests that maedi (or visna) virus is not implicated in the aetiology of pulmonary carcinoma of sheep.


Serological studies of mucosal disease virus in England and Wales. A survey of the cattle population in England and Wales showed that about 62 per cent of animals had neutralising (SN) antibody to BVD-MD virus in their serum. Fewer seropositive individuals were detected by immunodiffusion (53 per cent), and complement fixation (CF) tests (34 per cent). The prevalence of antibody was not affected by herd size, but some variation was found between various regions of the country, and the proportion of animals seropositive rose markedly with age. The immunodiffusion test was a specific but relatively insensitive method of detecting antibody compared with the SN test. The data suggest that the CF test may detect a different, short-lived antibody. For diagnostic serology, the SN test was superior to the other methods. No antibody to the virus was detected in human or equine sera but about 7.3 per cent of sheep sampled had serum antibody to the virus in the immunodiffusion test.


Increased serum gastrin concentration in hemodialysis patients treated with 1-alpha-hydroxy vitamin D. Serum gastrin and serum calcium concentrations were measured in 9 hemodialysis patients treated with 1-alpha-hydroxy vitamin D3 and in 15 control patients. Serum calcium and serum gastrin concentrations were significantly increased after 1 and after 5 months of treatment with 1-alpha-hydroxy vitamin D3. It is concluded that serum gastrin concentration is sensitive to a rise in calcium concentration within physiological range in patients with chronic renal failure.


A controlled comparison of corticotropin and hydrocortisone in the treatment of severe proctocolitis. A randomized controlled trial has been made of corticotropin (ACTH) 80 units given intramuscularly daily and hydrocortisone 400 mg infused intravenously over 24 hours, in the treatment of in-patients with severe proctocolitis. Seven patients were treated with ACTH and nine with hydrocortisone over an initial seven days. Three patients in the ACTH group and six in the hydrocortisone group improved and were discharged on medical treatment. Four patients treated with ACTH and two with hydrocortisone required urgent colectomy during this admission. Plasma cortisol levels were measured, but there was no correlation between these and the observed therapeutic response. Sideeffects were troublesome in both groups but were more marked in those treated with hydrocortisone. It is concluded that ACTH is not intrinsically superior to hydrocortisone in these doses, in the treatment of severe proctocolitis.