Proteins that affect DNA conformation.

Proteins that affect DNA conformation. In eucaryotic cells virtually all of the DNA is complex with proteins to form a unit fiber approximately 100 A in diameter. Chromatin is formed by the higher order coiling of the unit fiber. In procaryotic cells, as exemplified by E. coli, the actual structure of the chromosome is less clear (218), but the discovery of the DNA gyrase raises the possibility that the DNA helix in the cell is maintained in an underwound state. It may be important to consider these structural features of DNA in future biochemical studies on replication, transcription, and recombination. The recent discoveries of the DNA swivelases, the DNA gyrase, and the DNA unwinding enzymes considerably increase our knowledge of DNA biochemistry. As more is learned about these enzymes and their interaction with DNA, the prospects for understanding the details of DNA transcription, DNA recombination, and particularly DNA replication appear to be good.


[Comparative analysis of the action of an interferon inducer in in vitro and in vivo systems]. Investigation of interferon inductor (poly IC) in vitro (chick embryon fibroblasts) and in vivo (mice) showed that the main parameters of the preparation effect, i. e. induction of interferon and development of resistance to viruses are as a whole quite comparable. The phenomenon of hyporeactivity in the both systems was reproduced on repeated use of the inductor. At the same time significant differences in the stimulating effect of DEAE-dextran were registered. The merits and demerits of the in vitro system for using in studies on antiviral drugs of the above type are discussed.


Antiviral effect of commercial juices and beverages. Nineteen commercial juices or beverages were tested for inactivation of poliovirus type 1. Grape and apple juices and tea were particularly antiviral. Although antiviral in aqueous solution, ascorbic acid was ineffective after addition to juices.


Mechanism of nicotine-induced release of norepinephrine from adrenergic nerve endings: is generation and propagation of impulses necessary? The mechanism of release of 3H-norepinephrine (3H-NE) from adrenergic nerve endings by nicotine was investigated. Tetrodotoxin (5 X 10(-7) g/ml) abolished the contractile response of adrenergically innervated tissues, namely, rat isolated vas deferens, cat splenic strips, guinea-pig atria to transmural stimulation but not to nicotinic agents, and it did not affect the release of 3H-NE by nicotine from rat isolated vas deferens. Incubation of tissues in low Na+ (23 mM) and high K+ (50 mM) medium, which is known to inhibit propagation of nerve impulses, did not affect the nicotine-induced outflow of 3H-NE. These observations support the contention that the generation of action potential in the postganglionic sympathetic nerve fibres is not the cause of release of norepinephrine by nicotinic agents.


Late onset of fatal cytomegalovirus infection after renal transplantation. Primary or reactivation infection? Cytomegaloviurs (CMV) infections are a recognized problem in the first six months after renal transplantation. Studies have suggested that primary infections produce symptomatic disease, whereas reactivation infections are usually asymptomatic. Two patients are described who developed fatal CMV infections in the second year after transplantation. Both patients had typical CMV disease with fever, pneumonitis, and hepatitis. Results of serologic studies in one patient were characteristic of primary infection, with seroconversion at the time of disease and appearance of specific IgM antibodies.  The other patient had a similar antibody response at the time of his illness, but serial antibody tests showed that he had had a transient seroconversion earlier, in the second month after transplanation, that was not associated with clinical symptoms. These patients indicate that CMV infection must be considered in the differential diagnosis of serious febrile illnesses even in the late posttransplantation period and may occur either as the result of primary or reactivation infection. Serologic studies at the time of illness may not allow distinction between the types of infection.


Endorphins in psychiatry: an overview and a hypothesis. This article presents an overview of the biochemistry, pharmacology, and physiology of endogenous opioid peptides (endorphins). Clinical psychopharmacology of exogenous opiate agonists and antagonists is reviewed. The evidence presented in the review is compatible with a hypothesis that the level of functional endorphins may be related to psychological events, with a normal level needed for psychological homeostasis. One corollary of this hypothesis is that the level of opioids in the brains of the mentally ill may be disturbed. Therapeutic implications of this hypothesis are considered.


Peptides and psychoneuroendocrinology. A perspective. Research indicates that brain peptides exert both behavioral and endocrinologic effects in humans and animals. This review summarizes the best known behavioral actions of four endogenous peptides: luteinizing hormone-releasing hormone (LHRH), adrenocorticotrophic hormone (ACTH), vasopressin, and angiotensin. The hypothalamic-releasing hormones play a role in modulating pituitary-end organ systems. Behavioral disorders may, in the future, be susceptible to formulation in terms of changes in brain peptides. Peptide research in psychiatry may be approached in several ways.


[Clinical endocrine investigations in patients with pituitary adenoma (author's transl)]. LH-RH tests of hypothalamo-pituitary gonadotropinfunction were performed in 26 patients with pituitary adenomata before and after prolactin suppression and/or surgical treatment. Various degrees of galactorrhea together with menstrual disorders were the key symptoms in all patients. There was a significant association of elevated prolactin levels with impaired LH-responses. Seven patients have undergone transspenoidal dissection of the pituitary adenoma and in five of them the pituitary LH-response reverted to normal (R2) after surgery. Nine of 15 amenorrhoic patients had a recurrence of menstrual cyclicity, partly biphasic within 3 months and a normalized LH-response after Bromocriptin therapy. 19 of our patients were desirous of pregnancy. Five pregnancies were confirmed within 4 months of treatment.


Species difference in the secretory response to dopamine in the pancreas of dogs, cats, rabbits and rats. The effects of dopamine and L-dopa on the exocrine secretion of the pancreas were compared in rats, cats, rabbits and dogs. Dopamine and its precursor, L-dopa, induced a secretin-like stimulation in dogs, while their effects were significantly less in rats. The dopaminergic receptor stimulus was involved in the exocrine secretion of the pancreas in dogs, whereas in rats, the beta-adrenergic receptor was involved. Spontaneous secretion in the resting state was present in rabbits but absent in cats. This spontaneous secretion in rabbits could be definitely inhibited by either dopamine or L-dopa. The secretin-induced secretion in both rabbits and cats was markedly inhibited by either dopamine or L-dopa. Species difference in the secretory response to dopamine in the pancreas of dogs, cats, rabbits and rats may be ascribed to the difference in the incorporation of exogenous L-dopa and dopamine in the exocrine cells of the pancreas.


Morphological and morphometric studies on the terminal boutons on the neurosecretory cells of the rat paraventricular nucleus. Ultrathin sections of the male rat hypothalamus, including the paraventricular nucleus prepared by perfusion fixation, were observed with a transmission electron microscope. The neurosecretory cells were classified into four types based on the difference in the structures of cell organelles and in the number and appearance of secretory granules present. This was reported elsewhere (Yukitake et al. Cell Tis. Res. 177: 1-8, 1977). Morphological and morphometric analyses on the terminal boutons abutting on the surface of the neursecretory cells were performed. The boutons were classified into three types: first type, having only small clear vesicles; second type, having large cored vesicles; and third type, having closely packed small vesicles. The incidence of the occurrence of each type of terminal bouton as well as the ratios of the surface of the perikarya of the neurosecretory cells covered by the terminal boutons were almost constant in all types of neurosecretory cells. As the stage of the secretory cycle proceeded, the numerical densities of the small synaptic vesicles in the terminal boutons clearly decreased. These results were consistent with the idea that each type of neurosecretory cell represented a different stage in the cell specific secretory cycle rather than a difference in neuron grouping, and that the terminal boutons probably transmitted some information regulating the secretory function of the cells.


GABA in pancreatic islets.

GABA in pancreatic islets. A high concentration of gamma-aminobutyric acid (GABA) and a high activity of glutamate decarboxylase (GAD), enzyme for GABA synthesis, were demonstrated in the rat pancreatic islets using a recently developed assay method combining the GABase system with the enzymatic NADP-NADPH cyclin method. Both GABA concentration and GAD activity in the pancreatic islets were comparable to those in the central nervous system (CNS). An insulinoma consisting mainly of B cells possessed a high concentration of GABA and a high activity of GAD. On the other hand, in the pancreatic islets of rats after treatment with streptozotocin, a specific attacker of B cells, GABA was negligible in amount. The occurrence of GABA in the pancreatic islets, at least B cells, apparently supports the view of Fujita that the islet cells are paraneurons. Presence of exogenous GABA does not influence the glucose- and amino acid-induced insulin release. The uptake of 3H-GABA into the pancreatic islet was small in amount as compared with that into the brain tissue such as the substantia nigra. These observations suggest that the different compartmentation of GABA in the islets from that in the CNS.


Early signs of oral and inhalative cadmium uptake in rats. Female wistar rats, 170--190 g, were exposed for 90 days to cadmium oxide aerosols containing 25 and 50 microgram Cd/m3 and for 63 days to 100 microgram Cd/m3. Simultaneously female wistar rats, 170--190 g, were fed 25, 50, and 100 ppm cadmium in drinking water for 90 days. After inhalation and ingestion of the metal, there were comparable kidney cadmium levels, but higher liver and blood levels after oral uptake. Coincident with the higher blood cadmium concentrations, proteinuria was observed only after oral administration. Likewise, there was a significant decrease of serum iron after ingestion and no lowering of the serum iron after inhalation of the metal. The inhalation led to a marked dose dependent weight increase of the lungs, which was followed by an impairment of gas exchange. Obviously, after inhalative cadmium uptake of 90 days pulmonary changes precede renal damage.


Stimulation of antibodies to Epstein-Barr virus (EBV) in acute viral infections. Acute and convalescent sera from 77 patients with serologically confirmed influenza, measles and adenovirus infections and from 36 healthy controls were tested for the level of antibodies to Epstein-Barr (EB) virus. In  the three groups of patients significantly higher titers of antibodies to EB viral capsid antigen (VCA) were found as compared to the controls. In 19 patients twofold or higher rise in antibody titers between the first and second blood sample was demonstrated. It is suggested that in patients with influenza, measles or adenovirus infections, involvement of lymphocytes leads to reactivation of EB virus and antibody formation is stimulated.


Gel double immunodiffusion studies with sex human rhinoviruses. Rabbits were immunized on six occasions during a seven month period with fluorocarbon and sucrose denstiy gradient purified preparations of human rhinovirus types 1A,2,3,4,9, OR 14. Sera collected 1 week after the final immunization formed 1 or 2 preciptin lines when reacted by immunodiffusion against fluorocarbon purified preparations of each homologous immunizing virus. Heterologous preciptin cross reactions were detected between: RV1A antigen and rabbit sera to RV2, RV9 and RV14; RV2 antigen and sera to RV1A; RV14 antigen and sera to RV3. The heterologous "group" or C-antigenic nature of the cross reactions was suggested by the merging of heterologous preciptin lines formed against rabbit sera with "group" antibody preciptin lines formed against human sera and by the location of heterologous reaction close to the antigen well. In addition, the presence of C-antigenic particles in the fluorocarbon treated RV2 preparation was suggested by the demonstration that a subpopulation of viral particles migrated to a pH of 4.4 by isoelectric focusing.


Cell cycle-dependent multiplication of avian adenoviruses in chicken embryo fibroblasts. Propagation of CELO virus employing confluent monolayers of chicken enbryo fibroblasts (CEF) yielded virus titers one to two logs lower than those from confluent chicken kidney (CK) cells. An enhancement of virus production in CEF as measured by plaque formation was obtainedby infectng cultures in the growing non confluent state. Measurements of 3H-thymidine incorporation revealed a positive correlation between the DNA synthesis of CEF cultures at the time of inoculation and the amount of progeny virus, whereas  in the CK-CELO-system no such relation was observed. Requirement of replicative fibroblasts for CELO multiplication was also demonstrated by comparison of virus replication in synchronized stationary and serum stimulated CEF cells. In stationary CEF cells arrested in the G1 phase of the cell replication cycle by serum deprivation and infected withe CELO virs, no cytopathic effect could be observed, and only very low amounts of virus were produced. But 24 hours after release of these cells for growth by serum stimulation a logarithmic rate of virus multiplication and a complete CPE occurred. Infection of synchronized CEF cultures at different stages of the cell cycle revealed that CELO multiplication was correlated with the S phase of the infected cell. In synchronized CELO infected CEF cultures viral DNA synthesis started 12 to 14 hours after growth stimulation when cells were near the end of the S phase. In contrast, no viral DNA synthesis could be measured in growth arrested CELO infected CEF cells, when cellular DNA synthesis was low. Therefore not only production of infectious virus but also viral DNA synthesis is correlated with events during the S phase of the infected CEF cell.


Effect of treatment with certain flavonoids on Mengo virus-induced encephalitis in mice. Among the four flavonoids tested quercetin and morin proved to be significantly effective against lethal Mengo virus-induced encephalitis in mice when the drugs were adminsitered orally (p.o.). With subcutaneous (s.c.) administration all four drugs failed to prevent mortality in the infected mice. Quercetin produced maximum protective response in intraperitoneally (i.p.) or intranasally (i.nas.) infected mice when administered twice daily at doses of 20 mg/kg for a period of not less than four days. Single injections of the full daily dose of drug failed to prevent deaths in mice. Treatment must be begun at the time of, or prior to, virus inoculation. Delayed initiation of treatment was ineffective in preventing mortality.


[Submicroscopical study of the granule-containing cells of bronchial carcinoids]. The submicroscopical structure of three bronchial carcinoids was studied. In all the cases, in the cytoplasm of tumor cells secretory granules formed from cisterns of the Golgi apparatus were found. Two types of granules were observed. The first ones were from 150 to 250 nm in size, the second--from 400 to 600 nm. Granules less than 500 nm in size comprise about 90% and cannot be observed by light microscopy because of the low resolving power of the microscope. It is suggested that the secretory product is eliminated by diffusion through the granular and then plasma membrane. The granules are assumed to be heterogeneous in their chemical composition. Secretion of other compounds in addition to serotonin is presumed. The ultrastructural similarity of the tumor cells and Kulchitsky cells is no proof of theif histogenetic relationship.


Epstein-Barr virus-associated antigens in nasopharyngeal carcinoma. Thirty-five sera from American patients with nasopharyngeal carcinoma were examined for Epstein-Barr virus (EBV)-associated antigens and compared with 85 sera from patients with other head and neck cancers, 80 sera from patients with lymphoma, and 47 sera from healthy control subjects. There was a definite correlation between the presence of nasopharyngeal carcinoma and the level of antibody titers to EBV. In particular, two tests that detected antibody to early antigen and antibody to viral capsid antigen in the serum IgA fraction were highly specific for the presence of nasopharyngeal carcinoma. There was a significant decrease in these antibody titers with clinical remission of the disease in treated patients with nasopharyngeal carcinoma. Clinically, these tests should have important application in the management and follow-up of patients with nasopharyngeal carcinoma.


Cellular mixed tumors of the salivary glands. Mixed tumors are unusual in that they possess both epithelial and mesenchymal elements. The proportions of the two elements vary greatly; the lesions with a pronounced preponderance of epithelial tissue have been called cellular mixed tumors. This increase in epithelial cellularity often results in misdiagnosis or in concern that the tumor may be more aggressive than the ordinary mixed tumor--possibly even malignant. We have reviewed 1,095 consecutive parotid tumors that were removed at the Mayo Clinic, Rochester, Minn, from 1950 through 1970. Ninety-three of the lesions were diagnosed initially as cellular mixed tumors, and slides were available for review. Of these 93, 43 fulfilled our criteria of having greater than 80% of each tumor composed of the packed epithelial cells. All tumors had a corresponding reduction in the mesenchymal portion. The records of the 43 patients were studied to determine the clinical behavior of these neoplasms, particularly with regard to recurrence, metastasis, and malignant transformation. Our findings confirmed our clinical suspicions that some of these cellular mixed tumors, notably those that showed histopathologic evidence of an increased rate of mitotic activity, can and do act in a more aggressive manner.


Synovial sarcoma of the neck associated with previous head and neck radiation therapy. Synovial sarcoma is a rare neoplasm that uncommonly arises in the neck. Fourteen years after facial and neck radiation therapy for acne, synovial sarcoma of the neck developed in a young man. Possible radiation-induced benign and malignant neoplasms that arise in the head and neck region, either of thyroid or extrathyroid origin, remain a continuing medical problem.


Macrophages and resistance to tumours: influence of agents affecting macrophages and delayed-type hypersensitivity on resistance to tumours inducing concomitant immunity. Early, specific concomitant immunity to each of four tumours was inhibited by treatment with silica or carrageenan. Late, non-specific concomitant immunity was, with one exception, not inhibited by these agents. Treatment of non-immune mice with silica at certain critical periods before challenge promoted the growth of four of six syngeneic methylcholanthrene-induced tumours in their feet. Treatment with carrageenan was much less effective. Early and late concomitant immunity were inhibited by one or more agents inhibiting delayed-type hypersensitivity: irradiation, niridazole and reserpine. Irradiation of non-immune mice did not effect the growth of tumours in their feet. Treatment of non-immune mice with niridazole or reserpine actually inhibited the growth of some tumours. It is suggested that (a) mice offer some natural resistance to tumour growth, macrophages perhaps being effectors; (b) some tumour isografts may survive only if an inflammatory reaction occurs; (c) mechanisms akin to those of delayed-type hypersensitivity operate in the expression of concomitant immunity; (d) macrophages are important in early, specific concomitant immunity, but perhaps less so in the late non-specific phase.


A testosterone-secreting adrenal cortical adenoma. A virilizing, gonadotrophin-responsive adrenal cortical adenoma was removed from a 54-year-old woman. Following removal of the tumour, which contained and secreted androgens and estradiol, serum gonadotrophins rose to menopausal levels and the patient experienced menopausal symptoms. Pre-operative attempts to localize the source of androgens were unsuccessful.


Pathogenesis of fowlpox in laying hens.

Pathogenesis of fowlpox in laying hens. Egg production dropped after hens were inoculated with fowlpox virus originally isolated from a natural mild infection. The drop started from the 2nd week and continued to the 5th week after inoculation. All birds developed focal lesions at the site of inoculation, and some developed secondary lesions. The drop in individual birds was related to the severity of lesions. Birds challenged with fowlpox virus fifteen months after vaccination with pigeon pox vaccine were susceptible.


Poxvirus infection in a spectacled Amazon parrot (Amazona albifrons). Avian pox was determined to be the cause of death of a spectacled Amazon parrot (Amazona albifrons). Intracytoplasmic inclusion bodies were visualized by light microscopy in esophageal and bronchial epithelial lesions. Electron microscopy revealed pox-virus virions in the inclusions.


Reexpression of HPRT activity following cell fusion with polyethylene glycol. Polyethylene glycol-1000 (PEG-1000) induced fusion of HPRT (E.C. 2.4.2.8) deficient Chinese hamster cells with alpha-galactosidase A (E.C. 2.3.1.22) deficient cells from a patient with Fabry's disease yielded hybrids which contained both human and hamster HPRT, G6PD (E.C. 1.1.1.49), and APRT (E.C. 2.4.2.7) and Chinese hamster alpha-galactosidase B. Thus PEG-1000 mediated somatic cell fusion led to reexpression of Chinese hamster HPRT. It did not restore the expression of human alpha-galactosidase. Since PEG-1000 treatment of HPRT- Chinese hamster cells in the absence of human cells yielded no HPRT+ cells, it is concluded that the element responsible for the restoration of rodent HPRT was contributed by the human cells and not by the agent employed to promote fusion.


Demyelinating peripheral neuropathy in a guinea hen associated with subacute lead intoxication. In mammalian species lead acts primarily as a neurotoxin. Lead poisoning is common in avian species but lesions in the nervous system have not been associated with it. This paper describes a peripheral neuropathy with fragmentation and segmental demyelination of fibers in the sciatic nerves of an adult guinea hen (Numida meleagris) which died of lead poisoning.


Effect of avian adeno-associated virus on pathogenicity of Tipton virus in chicks. The avian adeno-associated virus (A-AV) reduced the pathogenicity of an adenovirus infection in vivo. Groups of chicks were infected with Tipton virus alone or in combination with high or low doses of A-AV. In both trials, the associated virus delayed and reduced chick mortality. This effect was dose-dependent and significant at the higher dose level.


Cigarette smoking and HDL cholesterol: the Framingham offspring study. High density lipoproteins were measured on fasting blood samples from 4107 men and women. Control for reported alcohol consumption and subscapular skinfold thickness using multiple regression analysis allowed an examination of the relationship between cigarette smoking and HDL cholesterol. Cigarette smoking was found to be associated with an average difference in HDL cholesterol of about 4 mg/dl in men and 6 mg/dl in women. Furthermore, when heavy alcohol drinkers were eliminated a significant negative association between number of cigarettes smoked and HDL cholesterol was demonstrable in both men and women. There was no evidence that former cigarette smokers, with the exception of those who switched to cigars or pipes or had quit less than one year, had lower HDL levels. Cigar or pipe smokers who had never smoked cigarettes had alcohol- and skinfold-adjusted HDL cholesterol comparable to the non-smoker. These observations indicate another possible link between inhaled tobacco smoke and the atherosclerotic process and suggest the need for further studies and experiments that might clarify the mutual relationship of HDL cholesterol, cigarette smoking and the atherosclerotic process.


Pharmacological profile of BR-931, a new hypolipidemic agent that increases high-density lipoproteins. BR-931 [4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio-(N-beta-hydroxyethyl)-acetamide], a new hypolipidemic agent of low toxicity, was evaluated in several tests of lipolysis and hyperlipidemia in rats, and in the cholesterol-induced atherosclerosis in rabbits. Significant hypolipidemic activity was observed in rats with doses of the agent at 12.5--50 mg/kg. In the Triton-induced hyperlipidemia, 50 mg BR-931 per kg was equieffective as 200 mg of clofibrate (CPIB) per kg. In contrast with CPIB, BR-931 exerted a powerful antilipolytic activity against epinephrine, ACTH, nicotine and cold exposure. BR-931 was particularly effective in diet-induced hyperlipidemias. Ethanol lipemia was totally prevented by the agent at 100 mg/kg. With Nath's diet, doses as low as 25 mg/kg significantly reduced hypercholesterolemia and hypertriglyceridemia. In these last two tests, the distribution of lipoprotein cholesterol was also determined. CPIB did not affect HDL cholesterol levels that had been decreased by the diets; in contrast, BR-931, already at doses of 50 mg/kg, brought the HDL/total cholesterol ratio back toward normal. A significant HDL cholesterol increase, together with some reduction of atheromatosis, was also observed in cholesterol-fed rabbits. BR-931,  a potent inducer of liver peroxisones and of mitochondrial carmitine acetyltransferase, appears to be a hypolipidemic agent of high efficacy and low toxicity for the clinical treatment of hyperlipidemias and atherosclerosis.


Studies on the metabolism of the lipoprotein Lp (a) in man. Lp(a) was isolated and labeled by reductive alkylation. Radioactivity only entered the protein moiety of the lipoprotein. No change in the immunological or physicochemical properties of Lp(a) was noted after the radiomethylation. After incubation of labeled Lp(a) with whole serum for 24 h in vitro, more than 99% of the radioactivity of the incubated sample was found in Lp(a). In 4 subjects Lp(a) was injected intravenously. A linear decline of the specific activity of Lp(a) in the serum was found when it was plotted semilogarithmically against time. Half-lives of Lp(a) in the serum were 35, 38, 53 and 58 h. In one subject, the "soluble" and the "insoluble" apoproteins of Lp(a) showed the same half-life as the whole Lp(a) molecule. This suggests that no exchange of Lp(a) apoproteins with lipoproteins of other density classes took place. At different times after the injection of Lp(a), 3--8% of the radioactivity of the serum was found in Lp B, and less than 2% of the radioactivity was detectable in VLDL and the fraction having a density of greater than 1.110 g/ml.


Acute effect of dietary therapy of type IV hyperlipoproteinaemia on the serum and lipoprotein concentrations and relative composition. In a detailed study the acute effect of diet and hospital admission on the plasma and lipoprotein lipid concentrations and composition was studied in 28 patients with type IV hyperlipoproteinaemia. Within 6 days there were significant falls in the mean serum cholesterol and triglycerides, very low density lipoprotein (VLDL) cholesterol and triglycerides and in high density lipoprotein triglycerides. These changes were accompanied by a significant rise in the mean low density lipoprotein (LDL) cholesterol concentrations. Using multiple regession models highly significant predictions of the change in VLDL triglyceride (R2 = 0.71) and LDL cholesterol (R2 = 0.47) were obtained utilising the pre-treatment lipoprotein levels as independant variables. Since elevated LDL cholesterol concentations are associated with atherosclerotic disease such models may have important therapeutic applications.


Lipoprotein composition in diabetes mellitus. Lipoprotein cholesterol and triglyceride levels have been determined in normal and diabetic Pima Indian women aged 20-35, HDL cholesterol levels were lower, LDL cholesterol levels were higher, and the ratio of HDL cholesterol/LDL cholesterol, a reflection of lipoprotein cholesterol distribution, was lower in the diabetics compared to the normals. VLDL triglyceride levels were also elevated in the diabetics. An analysis of lipoprotein composition suggested that these changes primarily reflect changes in numbers of particles, since lipid composition and lipid/protein ratios were similar in lipoproteins isolated from normals and diabetics. The ratio of ester/free cholesterol in LDL and HDL was lower in normal Pima Indians than in a comparable group of Caucasians, although plasma LCAT activity was not significantly different. The data indicate that diabetes may be associated with shifts in distribution of LDL and HDL, as well as with increases in VLDL.


Serum lipoprotein abnormalities in renal allograft recipients. Detailed lipid composition of serum and isolated lipoprotein fractions in male and female transplant recipients has been determined. Results have been compared with appropriate controls and a number of abnormalities have become apparent: (i) Serum total cholesterol, triglyceride and phospholipid levels were significantly elevated in transplant recipients. (ii) All lipid classes VLDL were significantly increased in both male and female patients. (iii) LDL-total cholesterol, triglyceride and phospholipid were significantly raised in female transplant patients, but this was true only for LDL-triglyceride in male patients. (iv) In female patients, HDL-total cholesterol and -triglyceride were significantly elevated, while in male patients, HDL-total cholesterol, but not HDL-triglyceride, was significantly greater than in controls. (v) The ratio of esterified to free cholesterol was significantly reduced in HDL of female transplant recipients. Results from correlation analysis suggest a relationship between some of the lipid abnormalities and renal function in female patients, while in male patients only immunosuppressive therapy is implicated.


Study of cultured skin fibroblasts from patients with and without ischemic heart disease. Metabolism of low density lipoprotein and cholesterol ester, synthesis of cellular lipids and effect of chloroquine on accumulation of cholesterol ester. The aim of the present study was to determine whether skin fibroblasts derived from patients with ischemic heart disease (IHD), which could not be related to accepted risk factors, would show a metabolic abnormality with respect to lipid or lipoprotein metabolism. Male patients 30-52 years old suffering from IHD were subdivided into two groups: those in whom IHD was not associated with risk factors such as hypertension, hyperlipoproteinemia, diabetes or smoking (group I); and those in whom heavy smoking was the only major risk factor recognized (group II). The controls were patients with angiographically normal coronary arteries (group III). Skin fibroblasts obtained from these patients were cultured and investigated with respect to metabolism of low density lipoprotein (LDL), synthesis of cellular lipids and induction of cholesterol ester accumulation in the presence of chloroquine, an inhibitor of lysosomal hydrolases. After 24 h incubation, the uptake and degradation of LDL protein in cells from patients of group II was significantly higher than in the controls, group III, but not different from those of group I. Hydrolysis of [3H] cholesterol linoleate, and incorporation of [3H] oleic acid into total lipids and into cholesterol esters was similar in cell cultures of the 3 groups studied. After exposure to chloroquine and LDL, the cells from the different donors accumulated cholesterol ester to a similar extent. Thus, whereas no significant difference was encountered in the lipid and lipoprotein metabolism in cells of patients with IHD without risk factors and controls, some increase in LDL metabolism was seen in cells from patients with IHD and with a history of smoking. It remains to be determined whether this increase was causally related to smoking.


Dyslipoproteinemia in patients with ischemic cerebro-vascular disease: a study of stroke before the age of 55. Serum lipoproteins were determined 8-12 weeks after the onset of ischemic cerebro-vascular disease (ICD) in 61 patients, 38 males and 23 females, before the age of 55. The results were compared with those of a matched control material. The diagnosis was based on clinical findings, CSF spectrophotometry, computer tomography, and angiography. Hyperlipoproteinemia was no common finding in these young and middle-aged patients with ICD. The normal mean total serum cholesterol concentration was the result of a slight increase in VLDL cholesterol and a concomitant HDL cholesterol reduction. In men, the HDL cholesterol concentration was lower than expected for any VLDL-TG concentration. The mean value of the HDL cholesterol concentration in the patients was 18% lower than in the control group. On agarose electrophoresis the lipoprotein variants "late prebeta", "sinking prebeta" and "rapid beta" lipoproteins could be demonstrated in the same frequency as in controls. There was no significant correlation between the degree of atherosclerosis, estimated by angiography, and any serum lipoprotein fraction. Several recent studies have stressed the importance of a low HDL concentration as an independent risk factor for atherosclerosis. The decreased HDL cholesterol levels found in the present material require further attention to the possible beneficial role of HDL in ICD.


Experimental hyper-beta-lipoproteinemia and its amelioration by a novel hypolipidemic agent. Experimental models for hyper-beta-lipoproteinemia were established in rats and the effects of certain hypolipidemic drugs were studied with these models. In the hyperlipemia induced in rats by feeding a high cholesterol diet, Y-9738 [ethyl 2(4-chlorophenyl)-5-ethoxy-4-oxazoleacetate] produced a dose-dependent reduction of serum cholesterol: such hypolipidemic activity was estimated to be about 7 times as great as that of clofibrate. On the other hand, clofibrate induced hepatomegaly at 100 mg/kg, whereas Y-9738 did not at this dosage, which is about 10 times the effective dose. Hyperlipemia induced by high cholesterol and thiouracil was characterized by increased beta-lipoprotein (heparin-calcium and disc electrophoresis). In this model, Y-9738 showed a dose-dependent lowering effect on beta-lipoprotein cholesterol with a marked decrease in the beta/alpha lipoprotein ratio. A tendency was noted for alpha-lipoprotein to be increased. In contrast, clofibrate exerted no effect on this hyper-beta-lipoproteinemia. These results suggest that the above models may be of value in exploring hyper-beta-lipoproteinemia and that Y-9738 may be more useful than clofibrate in the therapy of hyperlipemia.


Response kinetics of host and experimental solid tumour after adriamycin. The effects of adriamycin (Adr) on the solid-tumour model, hepatoma 3924A, and on critical organs of the host, were determined at intervals after single injections of 60 mg/m2 of the agent. A reduced rate of tumour growth was evident 4 days after treatment, continued to Day 11, and then returned to rates comparable to control values. On Day 11 tumour volumes of treated animals were 38% of control. During the period of reduced growth, 3H-TdR incorporation into tumour DNA and percentage labelled tumour cells were less than control values. DNA concentration in tumour was not affected by drug treatment, which differs from observations made in other studies employing 5-fluorouracil (FU). No evidence of significantly increased necrosis or fibrosis of the tumour was found after Adr. The Adr treatment caused loss of 60% of the tibial marrow by Day 4, as measured by total DNA content. Marrow DNA recovered to control levels between Days 7 and 11. Incorporation of 3H-TdR into heart DNA was reduced more than 40% during the first week after Adr treatment; enhanced incorporation was observed on Day 11, and control levels were attained by Day 17. No significant pathological evidence of cardiac toxicity was found 2-21 days after Adr but degeneration of myocardial cells and oedema was prominent at 14 weeks.


Effects of sensitizers on cell respiration: 1. Factors influencing the effects of hypoxic cell radiosensitizers on oxygen utilization of tumour and cultured mammalian cells. We have investigated the effects of 24 nitrocompounds, differing in their half-reduction potential, on the respiration in vitro of Ehrlich ascites tumour cells and cultured V79 lung cells. Many drugs with redox potentials more positive than--0.35 stimulated oxygen utilization in the presence of glucose. Glucose had little effect on the inhibition of respiration by drugs with oxidation-reduction potentials more negative than--0.38. Nitrocompounds that inhibited Ehrilch cell respiration in the presence of glucose, also inhibited intracellular reduction of ferricytochrome (c+c1). Drugs that stimulated oxygen utilization also stimuated intracellular (c " c1). Drugs that stimulated oxygen utilization also stimulated intracellular reduction of ferricytochrome (c + c1). A correlation between drug oxidation-reduction potential and stimulation of oxygen utilization in KCN-inhibited cells was found.


The neurotoxicity of misonidazole and its relationship to dose, half-life and concentration in the serum. Misonidazole has been given to a total of 87 patients. Neurotoxicity has occurred--convulsions with the higher doses and peripheral neuropathy with the lowern ones. The data from 34 patients receiving 4--7 doses has been analysed. Peripheral neuropathy is related to the total tissue exposure. By monitoring serum levels and controlling the total dose given, convulsions are avoidable and peripheral neuropathy restricted to a low and acceptable level.


[Acute intestinal amebiasis treated with aminosidine]. A clinical-therapeutic assay carried out at the "William Soler" pediatric teaching hospital in children admitted due to an acute diarrheic syndrome and treated with aminosidine sulfate is presented. Histolytic ameba is still an element to be considered in the origin of gastroenteritis. The therapeutic action of aminosidine is evaluated.


Peripheral neuropathy related to misonidazole: incidence and pathology. The human tolerance to multiple dosages of misonidazole (Ro-07-0582) was studied in 28 patients with different types of malignant neoplasias. The mean total dose for this group of patients was 16.2 g. The main toxicity was peripheral neuropathy with an overall incidence of 35%. This neuropathy occurred more frequently and with greater severity when the drug was administered 3 times a week and when patients received total doses of over 18 g. The best tolerated schedule appears to be once or twice a week up to total dosages of 18 g or less (approximately 11 g/m2). Electron microscopy of a sural nerve biopsy from an affected patient revealed residual of previous distal axonal degeneration, with some segmental demyelination and remyelination, which affected both large and small myelinated nerve fibres.


Further clinical experiences of a phase I study with the hypoxic cell radiosensitizer misonidazole. Since April 1976 we have performed clinical investigations with multiple doses of the hypoxic cell radiosensitizer misonidazole in 21 patients. A significant side effect of the drug was the development of peripheral sensory neuropathies in 13 patients (8 mild, 5 severe) and of a transient acute organic psychosyndrome in 2 of the 5 patients with a severe polyneuropathy. The severity of the polyneuropathies is related to the total dose of misonidazole and the overall time of drug administration. Treatment schedules designed to obtain the desired sensitizing effect without neurological side effects are under investigation.


The optimum regime for the administration of misonidazole and the establishment of multi-centre clinical trials. The amount of misonidazole which may be given to one patient is limited because of the risk of neurotoxicity. The drug may be given with every treatment of a multi-fraction course of radiotherapy or only with some of the treatments. The number of radiotherapy treatments can be reduced or multiple treatments given after each dose of the drug. Experience in 19 patients given daily doses of misonidazole is reported. This is practical regime to be considered for clinical trials of this hypoxic cell sensitizer.


Induction of some features of glial differentiation in primary cultures of human gliomas by treatment with dibutyrl cyclic AMP. Explants from 18 gliomas were cultured for periods up to 7 weeks and studied by light microscopy scanning and transmission electron microscopy. Well-differentiated tumor tissue gave rise to early outgrowths of stellate cells showing process orientation. Poorly-differentiated tissue produced a more haphazard out-growth of pleomorphic cells with few processes and flattened pseudopodia. Mean circadian cell displacement was several times greater in poorly-differentiated cells, but was significantly and reversibly reduced by treatment and dibutyryl cAMP (5 X 10(-4)M) for 48 h. Reduction in motility was directly correlated with a change in cell morphology to a more stellate form. Well-differentiated cells had a smooth surface with ruffling restricted to the ends of processes and highly orientated glial filament and microfilament bundles. The poorly-differentiated cell surface had a microvillous, blebbed appearance and ruffling regularly occurred around the edge of the cytoplasm. Glial filaments and microfilaments were fewer and less well orientated in the poorly-differentiated cells; sites of adhesion to the substratum were fewer than in well-differentiated cells. Treatment of malignant cultures with dibutyryl cAMP resulted in smoothing of the cell surface, retraction of processes into thin pseudopodia and the appearance of microfilament bundles within the cells. These features marked the apparent cyto-differentiation. However, there was a loss of attachment, disappearance of microtubules and loss of glial filaments in the cytoplasm which was not compatible with differentiation. Intracellular recordings of membrane potentials gave a significantly higher mean value for well-differentiated cells. The mean membrane potential and input resistance of poorly-differentiated cells was unchanged by the addition of dibutyryl cAMP. The results of this study suggest that some, but not all, features of mature glia are restored in malignant tumour cells by cAMP treatment.


Collagen prolyl hydroxylase levels in experimental paraquat poisoning. Collagen prolyl hydroxylase activity was measured in lung, liver, kidney and serum in rats killed on 7 successive days following a single toxic dose of paraquat. The enzyme activity in liver initially fell, then returned to normal, and no change was found in kidney. In contrast a 5-fold rise was observed in lung enzyme level, reaching of maximum on the 5th day. A parallel rise in serum activity was also found. The onset of these enzyme elevations corresponds to the early histological appearance of cellular proliferation in lung alveoli.


Primary mucinous carcinoma of the skin.

Primary mucinous carcinoma of the skin. Only twenty-seven cases of primary mucinous carcinoma of the skin have been reported. Three additional cases are included in this paper. The clinical presentation is distinctive; they are found most commonly in middle-aged, black males in the head and neck region as painless nodules, usually less than 3 cm in diameter. Previous reports have indicated the probable sweat gland origin of these tumours; one of our cases demonstrates a transition from a solid hidradenoma-like pattern to nests of tumour cells floating in mucin lakes, adding support for the sweat gland hypothesis. The clinical behaviour is relatively benign; late recurrences are common but metastases are rare. Only one case had widespread metastases. Although the histological appearance is distinctive, other primary sites first must be excluded. Local excision is the treatment of choice.


Translational recognition of the 5'-terminal 7-methylguanosine of globin messenger RNA as a function of ionic strength. The translation of rabbit globin mRNA in cell-free systems derived from either wheat germ or rabbit reticulocyte was studied in the presence of various analogues of the methylated 5' terminus (cap) as a function of ionic strength. Inhibition by these analogues was strongly enhanced by increasing concentrations of KCl, K(OAc), Na(OAc), or NH4(OAc). At appropriate concentrations of K(OAc), both cell-free systems were equally sensitive to inhibition by m7GTP. At 50 mM K(OAc), the reticulocyte system was not sensitive to m7GMP or m7GTP, but at higher concentrations up to 200 mM K(OAc), both nucleotides caused strong inhibition. The compound in m7G5'ppp5'Am was inhibitory at all concentrations of K(OAc) ranging from 50 to 200 mM, although more strongly so at the higher concentrations. Over the same range of nucleotide concentrations, the compounds GMP, GTP, and G5'ppp5'Am were not inhibitors. The mobility on sodium dodecyl sulfate-polyacrylamide electrophoresis of the translation product was that of globin at all K(OAc) concentrations in the presence of m7GTP. Globin mRNA from which the terminal m7GTP group had been removed by chemical treatment (periodate-cyclohexylamine-alkaline phosphatase) or enzymatic treatment (tobacco acid pyrophosphatase-alkaline phosphatase)  was translated less efficiently than untreated globin mRNA at higher K(OAc) concentrations, but retained appreciable activity at low K(OAc) concentrations.


Effect of specific lysine modification on the reduction of cytochrome c by succinate-cytochrome c reductase. The reduction of cytochrome c by succinate-cytochrome c reductase was studied at very low cytochrome c concentrations where the reaction between cytochrome c1 and cytochrome c was rate limiting. The rate constant for the reaction was found to be independent of ionic strength up to 0.1 M chloride, and to decrease rapidly at higher ionic strength, suggesting that the interaction between cytochrome c1 and cytochrome c was primarily electrostatic. The reaction rates of cytochrome c derivatives modified at single lysine residues to form trifluoroacetylated or trifluoromethylphenylcarbamylated cytochromes c were studied to determine the role of individual lysines in the reaction. None of the modifications affected the reaction at low ionic strength, but at higher ionic strength the reaction rate was substantially decreased by modification of those lysines surrounding the heme crevice, lysine-8, -13, -27, -72, and -79. Modification of lysine-22, -25, -55, -99, and -100 had no effect on the rate. These results indicate that the binding site on cytochrome c for cytochrome c1 overlaps considerably with that for cytochrome oxidase, suggesting that cytochrome c might undergo some type of rotational diffusion during the electron-transport process.


Modified nucleotides in T1 RNase oligonucleotides of 18S ribosomal RNA of the Novikoff hepatoma. The primary structure of 18S rRNA of the Novikoff hepatoma cells was investigated. Regardless of whether the primary sequence of 18S rRNA is finally determined by RNA sequencing methods or DNA sequencing methods, it is important to identify numbers and types of the modified nucleotides and accordingly the present study was designed to localize the modified regions in T1 RNase derived oligonucleotide. Modified nucleotides found in 66 different oligonucleotide sequences included 2 m62A, 1 m6A, 1 m7G, 1m1cap3psi, 7 Cm, 13 Am, 9 Gm, 11 Um, and 38 psi residues. A number of these modified nucleotides are now placed in defined sequences of T1 RNase oligonucleotides which are now being searched for in larger fragments derived from partial T1 RNase digests of 18S rRNA. Improved homochromatography fingerprinting (Choi et al. (1976) Cancer Res. 36, 4301) of T1 RNase derived oligonucleotides provided a distinctive pattern for 18S rRNA of Novikoff hepatoma ascites cells. The 116 spots obtained by homochromatography contain 176 oligonucleotide sequences.


Comparative studies on the 7-iodo and 7-fluoro derivatives of N-acetoxy-N-2-acetylaminofluorene: binding sites on DNA and conformational change of modified deoxytrinucleotides. GMP and native DNA were reacted with 7-iodo and 7-fluoro derivatives of N-acetoxy-N-2-acetylaminofluorene. It was shown that the 7-halogeno derivatives react on C-8 of guanine. Furthermore the respective amount of arylamidation (covalent linkage on the C-8 of guanine) and arylation (covalent linkage on 2-NH2 groups of C3 of guanine) addition products was determined in both native and denatured DNA-[14C]AAIF. Two G containing deoxytrinucleotides modified by either AAFF or AAIF were studied comparatively by means of circular dichroism, and as a function of several parameters known to affect the conformation of the deoxytrinucleotides. The induced optical activity in fluorofluorene ring seemed to be very sensible to the conformational changes of the deoxytrinucleotides. On the other hand, the AAIF residue exhibit a lower induced optical activity which remained unchanged when the deoxytrinucleotides conformation was affected. The results presented in this paper led us to conclude that the AAFF and AAIF modified deoxytrinucleotides adopt a conformation which nicely fits with the insertion-denaturation and outside-binding model, respectively.


Ferricytochrome c oxidation of cobaltocytochrome c. Comparison of experiments with electron-transfer theories. Electron transfer from cobaltocytochrome c to ferricytochrome c has been studied by stopped-flow kinetics. The second-order rate constant at pH 7.0, 0.1 ionic strenght, 0.2 M phosphate, and 25 degrees C is 8.3 x 103 M-1 s-1. The activation parameters obtained from measurements made between 20 and 50 degrees C are deltaHnot equal to = 2.3 kcal mol-1 and deltaSnot equal to = -33 eu. The rate constant is not significantly dependent on ionic strength; it is also relatively independent of pH between the pK values for conformation transitions. The rate diminishes at pH greater than 12. The self-exchange reaction of cobalt cytochrome c was investigated with pulsed Fourier transform 1H NMR. The rate is too slow on the 1H NMR scale; it is estimated to be less than 133 M-1 s-1. These results together with the self-exchange rates of iron cytochrome c [Gupta, R.K., Koenig, S. H., and Redfield, A. G. (1972), J. Magn. Reson. 7, 66] were analyzed by theories of Jortner and Hopfield. The theories predict the self-exchange of Cocyt c to be too slow for 1H NMR determination. The rate constant calculated by the nonadiabatic multiphonon electron-tunneling theory for the Fecyt c-Fecyt c+ and Cocyt c-Fecyt c+ electron transfers are in good agreement with experiments.


Characterization of phosphatidylthreonine in polyoma virus transformed fibroblasts. A threonine phospholipid in polyoma virus-transformed hamster embryo fibroblasts has been characterized as phosphatidylthreonine. The identification has been made by chemical and enzymatic hydrolysis. Acid hydrolysis of the phospholipid produces free threonine. Mild alcoholysis produces a water-soluble derivative having the properties of glycerophosphorylthreonine. Hydrolysis with phospholipase C produces phosphorylthreonine which on prolonged acid hydrolysis yields threonine. Phosphatidylthreonine in the cell is more accessible to reaction with fluorodinitrobenzene than is phosphatidylserine. Phosphatidylthreonine also has been found as a major aminophospholipid in two other polyoma-transformed hamster cell lines and in the BHK-21/C13 line including the PVT-3 and TS-3 lines. the latter derived from BHK cells. Only a trace amount of phosphatidylthreonine occurs in normal liver, kidney and spleen of the adult mouse, in normal liver and kidney of the adult hamster, in whole mouse and hamster embryos, and in mouse 3T3 cells and SV40-transformed 3T3 cells.


Electron capture at the iron-oxygen centre in single crystals of oxymyoglobin studied by electron spin resonance spectroscopy. Exposure of single crystals of oxy-myoglobin to 60Co gamma-rays at 77 K results in electron addition to the Fe-O2 units. The resulting ESR spectrum has been analysed to give the principal values and directions for the g-tensor of this unit. The results suggest that the dioxygen ligand is strongly tilted, the direction of tilt being close to one of the bisectors of the N-Fe-N bond angles. Possible reasons for this orientation are discussed.


Electron addition to the active site of Cancer magister haemocyanins. An ESR study of cu(II) centres after gamma-radiolysis. Exposure of aqueous glasses of Cancer magister haemocyanin to 60Co gamma-rays at 77 K results in a novel paramagnetic centre with ESR features showing hyperfine coupling to one strongly coupled 63/65 Cu nucleus and possibly one weakly interacting 63/65 Cu nucleus. Addition of electron scavengers showed that this centre is formed by electron addition. It is suggested that addition occurs at Cu2+-O2-Cu2+ units to give Cu2+-O2Cu+ centres. If this is correct, then electron-transfer between the two copper ions is slow or non-existent, possibly indicating that they are inequivalent. The centre is unstable, the signals being lost irreversibly on heating to approx. 270 K. The g parallel and A parallel (63/65Cu) data place this centre into the type I classification.


The reaction of mercaptans with tyrosinases and hemocyanins. 1. Titration of Neurospora tyrosinase with 2-mercaptoethanol shows that the increase of absorbance at 700 nm is directly correlated to the loss of enzymatic activity. Approximately 2 mol of 2-mercaptoethanol per mole of protein are needed for full development of the green, enzymatically inactive complex. The increase of absorbance at 700 nm is also proportional to the intensity of the EPR signal and the amount of non-covalently bound 2-[35S] mercaptoethanol to the enzyme. The maximal EPR intensity reaches 70% of the protein concentration and at most 0.7--0.8 mol of 2-[35S] mercaptoethanol is bound per mol of enzyme. 2. Stopped-flow measurements show that in the reaction between 2-mercaptoethanol and Neurospora tyrosinase a raction intermediate with a strong absorption band at 360 nm is formed in an apparent second-order reaction. This intermediate displays no EPR-detectable signals. The intermediate decays in a similar complex fashion as the absorption band at 700 nm is formed. 3. The reaction of Neurospora tyrosinase with a variety of sulfhydryl compounds was also investigated. In most cases green coloured, enzymatically inactive complexes are formed displaying slightly different EPR signals. However, with cysteine and cysteamine violet coloured, enzymatically inactive complexes are formed which show rather different EPR signals. The integrated EPR intensities amount to 40--70% of the protein concentration. Based on simulations of 9 and 35 GHz spectra all observed EPR spectra can be represented as true S = 1/2 systems. The cysteamine complex can be interpreted as arising from a mixed valence Cu2+ . Cu+ complex. The 2-mercaptoethanol spectra can, however, arise from sulphur radicals. 4. Treatment of Agaricus bispora tyrosinase and Cancer pagures hemocyanin with 2-mercaptoethanol results in green-coloured, EPR detectable complexes similar to the one found with Neurospora tyrosinase. No such complexes are formed when hemocyanins from Helix pomatia and Panulirus interruptus were treated with this reagent.


A calorimetric comparison of the interaction of sodium dodecyl sulfate with cytochrome c and erythrocyte glycoproteins. The interactions of sodium dodecyl sulfate with cytochrome c and erythrocyte glycoproteins have been studied by the method of titration calorimetry. It was found that the initial addition of sodium dodecyl sulfate to cytochrome c caused an endothermic unfolding of the protein, detectable by circular dichroism (CD). This was followed by the exothermic binding of sodium dodecyl sulfate to the protein, without further CD-detectable conformational changes. In contrast, sodium dodecyl sulfate bound directly to the erythrocyte glycoproteins in an exothermic reaction without any accompanying CD-detectable conformation changes. This indicates that the glycoproteins solubilized in aqueous media have exposed hydrophobic regions which can interact directly with this detergent. The enthalpy changes and stoichiometries of binding are reported.


[Binding of spin-labeled palmitic acid by bovine serum albumin]. Investigation of the binding of spin-labeled palmitic acid and its esters with bovine serum albumin is presented. Both probes are shown to bind strictly to strongly and loosely binding centers of protein. The quantitative study allows to determine the binding constant and number of protein binding sites. It is found that the total binding constant for esterified derivative is several times as much as that of palmitic acid.


Long-lasting effect of single ecdysone injections in newborn rats. Single injections of ecdysone to newborn rats were followed by increases in the weights of the adrenals and decreases in the weights of the thymuses in the adults. No change was registered in body weights and in the weights of the gonads. The plasma androsterone levels seemed to decrease; however, the interindividual differences proved to be too great to draw definite conclusions.


The conversion of aryl sulfate ester salts to alkyl aryl derivatives suitable for analysis by electron impact mass spectrometry. A series of monosubstituted (CH3, CH3O, Cl Br or NO2 at the O, m and p position) phenyl sulfate ester salts were derivatized to form n-propyl aryl sulfate diesters. The derivatization was accomplished by reacting aryl sulfate ester salts, AgClO4 and n-propyl iodide in SO2 at--40 degree C. The mass spectra of all the n-propyl aryl sulfate esters showed an intense molecular ion and intense diagnostic peaks at M--42 ([aryl--OSO3H]+) and M--122 ([ARYL--OH]+). The utility of this procedure for derivatizing selected sulfate ester conjugated xenobiotics and steroids was demonstrated.


[Photoinduced peroxidase reduction. Reaction with indolylacetic acid]. Kinetics of photoinduced reaction of HRP reduction by indolil acetic acid and dependence of the reaction rate and fluorescence of indoliol acetic acid on quenchers (I, Cst concentration was investigated. It was demonstrated that both I- and Cst action on the reaction differed from that on fluorescence. So the most probable reaction pathway is that through metastable triplet state of the substrate.


[Enzymes of tissue respiration and oxidative phosphorylation in the myocardium after neurogenic damage to it]. Three-hour electrical stimulation of the aortic arch in the rabbit myocardium is followed by a marked increase of the succinate dehydrogenase activity, by a sharp fall of the cytochrome oxidase activity and by uncoupling of the oxidation and phosphorylation processes. It is suggested that neurogenic myocardial damage is accompanied by discoordination in the energy processes regulation with uncoupling of the oxidative phosphorylation and by a decrease of the macroergic compounds synthesis.


[Effect of perturbing agents on the dynamic properties of immunoglobulin fragments]. The effect of NaCl, (NH4)2SO4, sodium dodecylsulphate and temperature on conformational properties of light-chain dimers and the Fab-fragments of immunoglobulins G has been investigated using the spin-label method. All these agents despite their different nature, induce qualitatively the same shift in the equilibrium between the A- and B-states of spin label towards the state of higher microviscosity A. The effect of 1.2 M NaCl and 0.75 M (NH4)2SO4 increase the stability of IgG in relation to temperature, as a result of this shift. The data obtained are interpreted in terms of the dynamic model of protein behaviour in water suggested earlier. In accordance with the model the action of the agents perturbing the conformation of protein and the ordered structure of water in the non-polar cavities of protein, reduce the life-time of this cavities in the water accessible state.


[Hepatic adenosine-3',5'-monophosphate metabolism in newborn rats irradiated during the period of organogenesis]. The adenylate cylcase (AC) and phosphodiesterase (PDE) activities, as well as the 3'5'-AMP (cAMP) fund were studied in the liver of newborn rats, intact, and after irradiation on the 9th day of the embryonic development. A decrease of AC and PDE activities was noted with a dose of 50 r. The stationary level of cAMP in the tissue remained unchanged. The adrenaline-stimulated AC activity only tended to decrease in case of irradiation. As suggested, during critical periods of development, in the presence of the hormone inductor in the liver of irradiated rats, conditions can be created for an increase of the cAMP pool.


[Spin glass type structures in chloroplasts]. The nature of discovered in [2] magnetic anisotropy of ESR signal with g approximately 3 appearing in chloroplasts at helium temperatures has been studied and clarified. It was found that a signal with these properties can arise only when dark adapted chloroplasts are cooled down to temperatures lower than 23 K in an external magnetic field (Ho greater than 2.300 Oe). The results obtained indicate that the system studied belongs to the type of "spin glasses", i. e., of the magnetically diluted structures with cooperative magnetic interaction between centres. These cooperative properties can be most distinctly observed at definite temperature and magnetic field strength values.


Platelet release reaction and intracellular cGMP. Enchanced cAMP concentrations inhibit the aggregation and release reaction of isolated human platelets and platelet-rich plasma to all known inducing agents. An opposing role for cGMP in this phenomenon has been proposed by some but not by others, and the function of cGMP in this secretory process is unclear. To further elucidate the role of cGMP in the release reaction, the effect of increased concentrations of this cyclic nucleotide on 14C-serotonin release was evaluated utilizing isolated human platelets and highly purified human thrombin or commercially available bovine thrombin. Several recently described stimulators of guanylate cyclase, including sodium nitroprusside, sodium azide, nitrosoquanidines, and ascorbic acid, were found to markedly augment platelet cGMP levels. Enhanced platelet cGMP concentrations produced by these drugs or by the exogenous addition of cGMP and its analogues neither caused these cells to secrete nor modulated the thrombin-induced serotonin release reaction. The inhibition of serotonin release by increased cAMP concentrations was not counteracted by increased cGMP levels. Platelet cGMP concentrations were unaltered by thrombin. These data indicate that cGMP is not an obligatory signal or a modulator of the thrombin-induced platelet release reaction.


[Contribution of immunohistochemistry to the cytological study of the adenohypophysis]. In the first chapter, the initial studies using immuno-histochemical methods for the cytological analysis of the adenohypophysis are presented; the different trends of the actual groups of research workers are summarized. In the second chapter are described, with details utillsation, some ser viceable techniques for experimental or anatomo-pathological studies of the pituitary gland; immunization of animals against hypophysial hormones, application to the study of the hypophysis (conditions of fixation, analysis of the subsequent times of histological techniques, immunohistochemical reactions [immuno-fluorescence and immuno-enzymatic reactions], controls of these reactions, applications on semi-thin and ultra-thin sections). In the third charpter are studied the results of this immunohistochemical analysis of the mammalian hypophysis: description of different cell types first in the anterior lobe then in the intermediate lobe; these observations are discussed. A special subdivision concerns the study of the human pituitary gland in normal and pathological conditions, in adults and foetuses. In the fourth chapter the results obtained in the study of the adenohypophysis of non mammalian Vertebrates: Fish, Amphibians, Reptiles, and Birds, are grouped. An extensive bibliography concludes this review (references until 1977). Illustrations, furnished by several french workers, are adapted to the different descriptions.


Treatment of oat cell carcinoma of the lung: complete remissions, acceptable complications, and improved survival. Oat cell lung cancer is a common disease which is usually disseminated by the time it is diagnosed. Treatment with cyclic combination chemotherapy (cyclophosphamide, doxorubicin, vincristine) administered concurrently with radiotherapy to the chest lesion and subsequent prophylactic brain irradiation was investigated in 36 patients with oat-cell carcinoma of the lung. Complete remissions occurred in 26 of the patients (15 of the 16 with limited-stage disease and 11 of the 20 with extensive-stage disease). Symptomatic improvement occurred in all patients. Twelve of the 16 patients with limited disease remained well and free of disease for over a year. The results were equivalent to those of a similar though more intensive regimen, but the toxicity was much less (there were no treatment-related deaths). Transient granulocytopenia with the risk of infection was the most serious complication. Survival and quality of life have been improved for all patients, particularly those with limited disease, who have previously responded poorly to treatment.


Recurrent genital herpes among women: symptomatic v. asymptomatic viral shedding. To investigate whether asymptomatic shedding of herpes simplex virus occurs in women with recurrent genital herpes, six women with documented disease were followed up twice weekly with viral cultures and pelvic examination. During the study period of 1330 patient days and 452 patient visits, 26 episodes of genital herpesvirus infection were recorded. Twenty-three (88%) episodes were accompanied by signs and symptoms. Three (14%) of the culture-positive recurrences were asymptomatic. In one episode an asymptomatic lesion was noted, and in two instances viral shedding from the vulva or cervix occurred in the absence of visible herpetic lesions. Three of the six women had evidence of recurrent cervical herpesvirus infection. No relationships between menstrual cycle and sexual activity and the onset of recurrence were noted.


Characterization of the retrograde transport of nerve growth factor (NGF) using high specific activity [125I] NGF. The process of the retrograde transport of nerve growth factor (NGF) has been recharacterized using a high specific activity preparation of[125I]NGF. Most of the general conclusions reached in the previous studies of Hendry, Thoenen and co-workers have been confirmed. However, significant quantitative differences were noted. Intraocular (anterior eye chamber) administration of[125I]NGF (less than 10 ng) resulted in accumulation in the superior cervical ganglia beginning at about 4 h. The ratio of radioactivity in the ipsilateral contralateral ganglia was 15--30:1. Maximal accumulation was seen at about 12h in the hamster and 16 h in rats. This pattern was quite different from that seen in other tissues. The uptake system from the eye of the rat was saturable (half-maximal at 15 ng) with maximal accumulation of 35--40 pg/ganglion. Systemic administration of[125I]NGF (200 ng) to adult rats resulted in no accumulation in SGG or celiac ganglion prior to 3 h, with subsequent rapid accumulation by 6 h and a rapid fall in radioactivity after 12 h. A similar time course was seen in 5-day-old rats, although the time curve was shifted slightly toward shorter time. The radioactivity in ganglia co-migrated with native NGF by SDS gell electrophoresis. Cytochrome c of comparable specific activity was not transported, and NGF did not stimulate the uptake and transport of cytochrome c. The retrograde transport of[125I]NGF was inhibited by the co-administration of biologically active, but not inactive, oxidized derivatives of NGF. By any route of administration, a significant percentage of the transported[125I]NGF was found in a purified nuclear fraction of the ganglia. Coupled with previous observations of specific nuclear NGF receptors in embryonic chick and sympathetic ganglia, this suggests that, after internalization and retrograde transport, NGF may directly act on the nucleus to produce at least some of its effects on the responsive cell.


Direct mapping of beta-adrenergic receptors in the rat central nervous system by a novel fluorescent beta-blocker. DL-N-(2-Hydroxy-3-napthyloxypropyl)-N'-dansylethylenediamine, dansyl analogue of propranolol (DAPN) is a novel fluorescent beta-adrenergic antagonist with high affinity to beta-receptors. The distribution pattern of DAPN fluorescence was studied in the rat central nervous system subsequent to its intravenous administration to living rats. DAPN distinctly labels specific regions and cells in the central nervous system (CNS). Highly dense DAPN fluorescence was observed in the pyramidal cell layer of the hippocampus, the granule cell layer of the dentate gyrus, the basal layers of the piriform cortex and the neocortex, the cerebellar Purkinje cell layer, and the spinal a-motoneurons. Pretreatment of control rats with DL-and L-propranolol markedly decreased the intensity and density of DAPN fluorescence in the tissue sections, whereas prior administration of D-propranolol had almost no effect. Pretreatment with large doses of reserpine did not alter the pattern of DAPN fluorescence. These findings were identical to those observed with another fluorescent beta-blocker, 9-aminoacridino-propranolol (9-AAP). Our data suggest that fluorescent beta-adrenergic antagonists may be used in vivo for the direct probing of the beta-receptors within the mammalian CNS.


Binding assay for opioid peptides with neuroblastoma x glioma hybrid cells: specificity of the receptor site. A sensitive and rapid radioreceptor assay has been developed to monitor opioid peptides in column effluents. It is based on competitive binding to NG 108-15 cells using [3H-Tyr]Leu-enkephalin as the displaced ligand. The specificity of binding to the cells of naturally occurring opioid peptides and synthetic analogs has been shown to be similar to that found with synaptic plasma membranes.


Electrophysiological demonstration of an excitatory subthalamonigral pathway in the rat. The subthalamonigral projection was studied with extracellular techniques in the rat. Orthodromic and antidromic stimulations of this pathway were performed in different sets of experiments in order to exclude the possible activation of passing fibers. A majority of the recorded cells in the subthalamic nucleus (STN) were antidromically activated by nigral stimulation. It was furthermore shown that the subthalamofugal fibers are excitatory to cells located in the pars compacta as well as in the pars reticulata of the substantia nigra. The possible existence of another descending STN pathway is also discussed.


Participation of limbic-hypothalamic structures in circadian rhythm of slow wave sleep and paradoxical sleep in the rat. The effect of brain lesion or surgical isolation of the neural circuit on SWS and PS circadian rhythm have been studied in female rats under a 14/10 light-dark schedule. Cortical EEG'S AND DORSAL NECK EMG were used to monitor SWS, PS and alertness in female rats. Intact and operated controls showed regular 4-5-day vaginal cycles and nocturnal sleep rhythm, but the night PS value on proestrus was lower than in other cycles. Following septal lesion, MPO roof cut, vaginal cycles and SWS rhythm were regularly maintained; however, the PS appearance at night, except during proestrus, increased (night PS peak). These results were similar to those for pinealectomized or ovariectomized female rats. A frontal cut of the MBH produced persistent estrus and disturbed both SWS and PS circadian rhythm. The suprachiasmatic-lesioned rats showed persistent estrus and disrupted SWS rhythm, but regularly maintained the circadian PS rhythm. The vaginal cycles and SWS rhythm in the fornical-transected rats were regularly maintained, but the PS rhythm was disturbed during diestrus and showed ultradian rhythm. From these results, it is suggested that the pineal hormone and the gonadal feedback mechanisms may be involved in the night PS peak and this mechanism may involve the septal- and amygdaloid-hypothalamic systems. A different neural mechanism exist for SWS and PS circadian rhythm; SWS rhythm involves the suprachiasmatic-basal hypothalamic system and PS circadian rhythm is related, in part, to the hippocampal-hypothalamic system.


The clearing of excess potassium from extracellular space in spinal cord and cerebral cortex. The relative importance of active and passive transport processes in the clearing of potassium released from active neurons was estimated Extracellular potassium activity [K+]0 was measured with ion-selective microelectrodes in the sensory area of the neocortex and in lumbosacral spinal cord of cats. Transient elevation of [K+]0 was evoked in cortex by stimulation of VPL and in spinal cord by stimulation of afferent nerves. The rate with which excess [K+]0 was cleared was either feebly or not at all influenced by variation of the intensity and frequency of stimulation. The half-decay times of [K+]0 were however prolonged when the duration of stimulus trains was increased. Only small differences were seen in the rate of decay of [K+]0 transients recorded at different locations within the gray matter; the shortest half-decay times occurred where K+ responses were largest. The different profiles of distribution of delta [K+]0 in response to stimulation of the cortical surface and of VPL nucleus were mapped. As in spinal cord also in cortex the distribution of the evoked sustained shifts of electric potential mirrored the distribution of [K+]0 transients. The rate at which K+ could diffuse out of volume sources similar in magnitude to the volumes of distribution of [K+]0 responses in gray matter were calculated. The observed half-decay times of [K+]0 transients were more than a hundred times shorter than those calculated for diffusion either in spinal cord or in cortex. Intravenous administration of digitoxigenin was shown to retard the clearing of [K+]0 and caused an elevation of the unstimulated [K+]0 baseline. Seizures were frequently induced by digitoxigenin when the [K+]0 baseline was only slightly elevated, and the occurrence of seizures was not associated with a definable threshold level of [K+]0. It is concluded that active reuptake is the principal mechanism of the clearing of [K+]0 released by neurons. Redistribution of K+ by diffusion must have been negligible under the conditions of these experiments, but may be more important when only a few neurons release K+ amongst many inactive cells. Considerations of a glial transport network are probably inconsequential for theories of the generation of seizures.


Lateral geniculate cell responses to electrical stimulation of the retina. Electrical stimulation of the retina evokes at the optic tract level rhythmic bursts of activity whose temporal structure is predictable from the polarity of the stimulation and the receptive field type. The reaction of lateral geniculate units to this input was studied in fast and slow relay cells as well as in interneurons. The results revealed that fast relaty cells presented a response whose temporal structure remained essentially unmodified in comparison to that observed at the optic tract level: that both anodal and cathodal polarities produced rythmic pattern of excitation the latency of which depended upon receptive field type and polarity applied. In slow relay cells and interneurons responses with equal latencies could be evoked for both polarities. Following cortical depression with 3 M KCl the latency of first bursts was unaffected in relay cells, while about one third of interneurons showed a temporal pattern which was similar to that recorded at the optic tract level after the treatment. This suggests that both ON and OFF retinal networks converge upon one geniculate slow P cell and interneuron, whereas fast relay cells are mostly driven by one of the two systems. Furthermore this convergence may be achieved through visual cortex in some units.


Electrophysiological changes induced by paradoxical sleep deprivation and lithium chloride poisoning in rats. In an attempt to analyze the disruption of conditioned taste aversion (CTA) caused by pre-acquisition paradoxical sleep deprivation (PSD), the effect of poisoning (0.15 M LiCl, 4% body weight) on the sleep--wakefulness pattern was studied in 12 rats with chronically implanted electrodes. The polygraphic recording showed that poisoning reduced the total sleep time in the subsequent 3 h from 57 to 42% and REM sleep from 6 to 2%. The lithium chloride effect was still more pronounced after 24-h REM sleep deprivation: the total sleep time decreased from 67 to 46% and REM sleep from 13 to 2%. The change of sleep--wakefulness pattern, most pronounced during the first hour of poisoning, gradually returned to normal but REM sleep was significantly reduced even 3 h after poisoning. It is concluded that 24-h PSD does not alleviate the subsequent poisoning. On the other hand, the lithium chloride induced reduction of post-acquisition REM sleep many enhance the learning impairment caused by pre-acquisition PSD.


Intracellular horseradish peroxidase injection for correlation of light and electron microscopic anatomy with synaptic physiology of cultured mouse spinal cord neurons. Synaptic interactions between spinal cord neurons grown in dissociated cell culture were studied electrophysiologically, and presynaptic cells were subsequently injected by intracellular iontophoresis with horseradish peroxidase (HRP). Following histochemical processing, injected cells were filled with dense reaction product which facilitated the light and electron microscopic identification of the individual physiologically typed neurons. This technique applied to neurons in monolayer culture allowed the visualization of complex intercellular relationships in essentially two dimensions. The number and distribution of morphologically defined synaptic contacts was determined for correlation with individual evoked postsynaptic potentials. HRP-filling of inhibitory and excitatory neurons revealed differences with respect to cellular geometry, axonal projection, and the number, location and ultrastructure of synaptic contacts.


Enzymatic and morphological properties of primary rat brain astrocyte cultures, and enzyme development in vivo. The development of (Na+ + K+) ATPase, carbonic anhydrase and HCO3--stimulated ATPase activity was studied in developing rat brain in vivo, and in primary astrocyte cultures from 1--3-day-old rat brain as a function of increasing cell growth. The primary cultures showed an increase in all the above enzyme activities during cell growth, with time courses which were qualitatively similar to their development in vivo. Cell cultures grown separately from the cerebellum plus brain stem regions showed greater carbonic anhydrase activity than cerebral cultures over the entire 4-week growth period, corresponding to development of this activity in these same regions in vivo, HCO3-stimulated ATPase activity was slightly greater in cerebellar cultures and (Na+ + K+) ATPase activity was greater in cerebral cultures up to the second week of growth, resembling development of the same enzyme activities in vivo. C6 glioma and neuroblastoma cells showed no and 10-fold lower carbonic anhydrase activities respectively, compared to the primary astrocyte cultures. Addition of 1 mM N6-2'-O-dibutyryladenosine-3',5'-monophosphate (DBcAMP) in the presence of serum caused marked formation of cellular processes and increased carbonic anhydrase and (Na+ + K+) ATPase activity. Maximum effects were found 2 h after addition of 1 mM DBcAMP and thereafter declined. In the absence of serum such effects persisted for at least 24 h. Electron microscope studies showed large numbers of microtubule (approximately 20 nm diameter) and filamentous structures (less than or equal to 10 nm diameter) in the cytoplasm, which showed changes in distribution in cells treated with DBcAMP. This study suggests that the increase in ATPase and carbonic anhydrase activities in rat brain with increasing age may be in part a reflection of proliferation and development of astroglia cells. Together with the morphological data, it also provides additional evidence that primary cultures derived from neonatal rats may closely resemble developing astroglia in vivo.


Prolidase activity in rat brain; developmental, regional and subcellular distribution. We determined the regional and subcellular distribution of prolidase in rat brain and its changes with development. The most rapid changes in enzyme activity occurred perinatally, with a maximum level of activity 2 days before birth and a minimum 1 day after birth. Of the 7 regions examined, cerebellum had the highest enzyme level, followed by medulla. The lowest levels were found in the hypothalamus in the adult and in the midbrain in the young. Polidase was mainly soluble; over 55% was recovered in the S2fraction, and the rest was released from the particulate fractions by hypotonic shock. Brains of male rats contained a slightly higher level of the enzyme.


Inhibition of enzyme induction in Pseudomonas aeruginosa by exogenous nucleotides. Alkylsufatase induction in resting cell suspensions of P. aeruginosa was inhibited by exogenously supplied adenosine or by ATP (2mM). Adenine phosphate had no effect while AMP or ADP caused a slight stimulation of induction. The inhibitory effect of ATP required the presence of added Mg2+, was not reversed by cyclic-AMP (2mM), and was independent of the nature of the inducer. Of a number of other nucleoside triphosphates tested, only UTP (2mM) acted as an inhibitor of induction. These nucleotides at external concentrations of 6mM also inhibited alkysulfatase induction in actively growing cells.


Congenital syndactyly: a reappraisal.

Congenital syndactyly: a reappraisal. Digital syndactyly is a common congenital anomaly and is associated with other anomalies affecting the digits, the hand, the arm or even the entire body. In the past it has been considered a single entity. However, recent information provided by experimental embryology, studies of morphogenesis and other sources, suggests that there are at least two distinct entities: primary syndactyly due to interference with the sequence of events that normally culminate in the division of digits into discrete parts and secondary syndactyly due to readhesion of adjacent digits as a result of close contact between raw surfaces. Primary syndactylyl may arise de novo, or it may occur as a manifestation of a mutation phenomenon or as a genetically controlled syndrome. Secondary syndactyly is the result of mechanical adhesion of adjacent parts involved in a general reparative or healing process. The initial insult leading to amputation of a portion of the digits may or may not be genetically controlled; adhesion is fortuitous. Management of syndactyly is determined by the complexity of the malformation. For simple cutaneous syndactyly the surgeon should make a zigzag incision and provide a rectangular, proximally based flap for the floor of the web, usually before the child is 2 years of age. A full- or split-thickness skin graft should be applied to the defects. For complex deformities involving several digits and associated with postural or osseous malformation, several operative procedures may be required. The surgeon must be vigilant to note and correct the sequelae resulting from recurrent contractures and imbalances associated with growth.


Combination modality therapy in lung cancer: a survival study showing beneficial results of AMCOF (adriamycin, methotrexate, cyclophosphamide, oncovin and 5-fluorouracil). Thirty-seven patients with advanced or recurrent lung cancer were randomized to cytoxan (CTX) alone, COMF (CTX, oncovin, methotrecate and 5-FU) or AMCOF (adriamycin, methotrexate, CTX, oncovin and 5-FU) after receiving radiation therapy to primary and bulky tumor sites. Median survival was 3 months for CTX, 6 months for COMF and 14 months for AMCOF. Analysis of those with cell (small cell) carcinoma showed median survival of 8.5 months. Oat cell cases treated with CTX survived 5 months (8 patients) with COMF 7.5 months (15 patients) and with AMCOF 13 months (14 patients). The median survival of those with adenocarcinoma or epidermoid carcinoma treated with CTX survived 3 months, with COMF 6 months and with AMCOF 15.5 months. Toxicity was moderate though no life-theatening toxicity developed in spite of the protocol design of escalation to achieve some degree of hematologic toxicity in all patients.


Intraaterial hepatic infusion and intravenous adriamycin for treatment of hepatocellular carcinoma: a clinical and pharmacology report. Four patients received intraarterial (ia) hepatic infusion and 10 received intravenous (iv) adriamycin for hepatocellular carcinoma. Four of each group are evaluable. The remaining 6 patients died within 14 days of intravenous therapy and are, therefore, considered nonevaluable. Patients received 2 to 9 courses of adriamycin every 3 weeks. One half of each group of evaluable patients had partial responses (pr). The group had pr for 22.5 weeks (range 8 to 37). The iv group had pr 27.2 weeks (range: 16 to 38.5). Mean survival was 21 weeks for nonresponders, and 43 weeks for responders. Intraarterial infusion did not protect patients from adriamycin toxicity. Cardiac and liver toxicity were not seen, but marrow and gastrointestinal toxicity developed at 1.2 X 10(-7)M adriamycin serum level. Adriamycin disappearance curves after ia and iv therapy were similar for similar bilirubin levels, and prolonged with hyperbilirubinemia. Ascites fluid did not accumulate detectable adriamycin. Pharmacokinetics are described in this report.


The effect of estradiol-17-phenylpropionate and estradiol benzoate on N-nitrosomorpholine-induced liver carcinogenesis in ovariectomized female rats. The influence of synthetic estrogens on the N-nitrosomorpholine (NNM)-induced liver carcinogenesis in ovariectomized young adult female rats was investigated and compared to rats which received only the carcinogen or estrogens. Estrogens when chronically administered after the cessation of carcinogen treatment increased the carcinogenic effect of NNM. In such conditioned animals the number of nodules per number of rats was 23/31, that of hepato-cellular carcinomas 9/31, whereas in animals which received only the hepato-carcinogen the incidence of nodules and carcinomas in liver was respectively 11/31 and 3/31. Higher incidence of benign and malignant tumors in other organs was also observed in these animals. Rats which received a single dose of estrogens simultaneously with NNM developed slightly fewer tumors in liver and in other organs. Since under my experimental conditions the long-term treatment with synthetic estrogens alone did not induce any focus, nodule or hepatocellular carcinoma in the liver. I suggest that the estrogens were acting rather as tumor promotors than true initiators of liver carcinogenesis.


Mucinous carcinoma of anal duct origin presenting clinically as a vaginal cyst. The vast majority of vaginal submucosal cysts are benign lesions. Primary malignant tumors of the vagina are infrequent and most are mucosal lesions. A case is described in which an unusual neoplasm, anal duct carcinoma, presented clinically as a vaginal lesion. The importance of considering anal neoplasia in the differential diagnosis of cystic vaginal lesions is noted.


Histiocytic tumors of the larynx: a clinicopathological study with review of the literature. A case of primary malignant fibrous histiocytoma of the larynx occurring in a 58-year-old man is described. This neoplasm is extremely rare in the larynx and the case reported (the second described by me) is the fifth so far reported in the world literature. The patient died after 19 months from first surgical treatment and the autopsy confirmed the histological diagnosis. The cases of histiocytic tumors of the larynx previously reported in the literature are re-examined and reclassified. Biological behavior and the therapy of the tumor, as well as differential diagnosis from other neoplasms, are discussed.


Malignant fibrous histiocytoma (malignant fibrous xanthoma: xanthosarcoma) of bone. Seven cases of malignant fibrous histiocytoma of bone are documented. In addition to typical histologic features such as the presence of a storiform pattern, histiocytic and fibrocytic cell types and severe cellular atypism, in five cases there were areas in which osteoclast-type giant cells were prominent and the lesion resembled a giant cell tumor. Three of the seven patients developed pulmonary metastases within two years of amputation, one developed extensive local recurrence including soft tissue and vascular invasion, and the other three are without evidence of disease but have been followed for only short periods.


Lobular neoplasia (so-called lobular carcinoma in situ) of the breast. In a review and reclassification of 5,560 benign epithelial lesions of the breast entered in the files of the Laboratory of Surgical Pathology at Columbia, we found 211 examples of the type of lobular proliferation occurring alone without co-existing infiltrating carcinoma, which we prefer to call lobular neoplasia, but which is generally referred to as noninfiltrating lobular carcinoma in situ. We regard this lesion as a separate distinctive pathological-clinical entity. These 211 cases are studied from a number of parameters, including the ages of the patients, the breast affected, the length of the follow-up, the interval between the initial diagnosis and the frank carcinoma which eventually developed in 17.1 percent of the patients. The relationship of microscopic qualitative and quantitative variations in the lobular neoplasia to subsequent carcinoma was studied; the variations were not found to have any value in predicting subsequent carcinoma. This study is unique in that we have data as to the frequency of a family history of carcinoma in a mother or sister, and also as to the occurrence of gross cystic disease in our patients with lobular neoplasia. We have determined the ratio between the observed and expected numbers of patients developing carcinoma in the several possible combinations of these three factors which predispose to carcinoma. We report that the predisposition is cumulative: in patients in whom all three predisposing factors were present the ratio of observed to expected risk of carcinoma was 13:8. We do not recommend mastectomy for lobular neoplasia, but only systematic follow-up by palpation of the patients' breasts every four months.


Appearance of intestinal type of tumor cells in hepatoma tissue induced by 3'-methyl-4-dimethylaminoazobenzene. Carcinoma tissues induced by 3'-methyl-4-dimethylaminoazobenzene were investigated both morphologically and biochemically. The most prominent histological pattern was an undifferentiated carcinomatous one. While this type of carcinoma, histologically, appeared to be due to a uniform population of cells, electron microscopic examination revealed that the carcinoma tissue was composed of many types of cells including cells that contained either the brush border or the mucous droplets seen in goblet cells. In addition, tumor cells that contain serotonin-like granules were noticed. An electrophoretogram of alkaline phosphatase in the tissue extract of this type of carcinoma revealed distinctly the presence of its intestinal isozyme. These findings evidently show that carcinoma induced by 3'-methyl-4-dimethylaminoazobenzene includes in addition to the cells differentiated toward hepatocytes or cholangiolar cells, those differentiated toward intestinal epithelial cells.


Lysosomal enzymes in macrophages of colonic tumors induced in rats by 1,2-dimethylhydrazine dihydrochloride. Ten weekly doses of dimethylhydrazine (30 mg/kg) were given to rats to induce colonic tumors. Histochemical and electron cytochemical studies revealed a distinct pattern of lysosomal acid phosphatase and beta-glucuronidase activity in macrophages in the stroma of these neoplasms. A dramatic increase in the number of acid phosphatase-rich macrophages was present in adenomas when compared to that in normal colonic mucosa. Fewer numbers of these cells were seen in well-differentiated adenocarcinomas, and they were barely detectable in highly invasive mucinous adenocarcinomas. It is postulated that these macrophages may play a role in preventing the invasion of adenomatous neoplasms into the submucosa. Application of histochemical techniques to study macrophage lysosomal enzymes may prove a useful diagnostic tool in differentiation of human colonic tumors for prognostic evaluation.


Aggregation of platelets and cell membrane vesiculation by rat cells transformed in vitro by Rous sarcoma virus. Primary rat embryo cells and established normal rat kidney cells transformed in vitro by Rous sarcoma virus induced the aggregation of rat platelets in vitro. The aggregating activity was shown to be specific for the transformed cells and was absent in the normal parent cells. The aggregation reaction is accompanied by the release of serotonin from the platelets. Further analysis and purification of this activity from the transformed cells demonstrated that the activity is shed from the cells growing in culture and is associated with membrane vesicles of heterogenous size. The normal cells also produced vesicles in culture; however, the level of vesicle productio was less than that from transformed cells, and the platelet aggregation and serotonin release activities were greatly reduced or absent in these vesicles.


In vitro effects of lipoprotein-associated cytotoxic factor on rat prostate adenocarcinoma cells. As assayed by the colony inhibition technique, sera from rats at an advanced stage of pregnancy were cytotoxic to a number of tumorigenic and "nontransformed" cell lines. The cytotoxic effect is not species specific. Primary fetal rat kidney cells were not susceptible to this cytotoxic effect. The level of demonstrable cytotoxicity in sera rose gradually as gestation advanced; it peaked at 48 hr before parturition and disappeared thereafter. Sera from control nonpregnant rats were not cytotoxic. The cytotoxic factor(s) in sera was demonstrable only in the very low-density lipoprotein (VLDL) (hydrated density less than 1.006 g/ml) fraction. Reconstitution studies suggest that: (a) inhibitors which usually mask the cytotoxic actitivity of VLDL in whole serum, are present; and (b) pregnancy initiates enhanced VLDL synthesis, which exceeds the masking effect. The possible role of this cytotoxic VLDL in host defense against neoplasia is discussed.


Effect of phorbol myristate acetate on cyclic nucleotide levels in mouse epidermis. Basal levels of cyclic adenosine 3':5'-monophosphate and cyclic guanosine 3':5'-monophosphate were determined in mouse epidermis in vivo after a single topical treatment with the tumor promoter phorbol myristate acetate. No changes in cyclic adenosine 3':5'-monophosphate levels were found from 0 to 72 hr after treatment. A twofold increase in cyclic guanosine 3':5'-monophosphate was found 36 hr after treatment. This increase had subsided by 72 hr. The effect of phorbol myristate acetate on DNA, RNA, and protein synthesis in the epidermis of Ha/ICR mice was also measured.


Cultural, morphological, cell membrane, enzymatic, and neoplastic properties of cell lines derived from a Hodgkin's disease lymph node. A neoplastic cell line (designated HuT11) has been established in continuous culture from an involved lymph node of a patient with Stage IIA Hodgkin's disease of the mixed cellularity type. The HuT11 line has been morphologically heterogeneous, consisting of mononucleate lymphoid-like cells, polygonal epithelioid cells, and mono-, bi-, and multinucleate giant cells. Four clones initiated from isolated binucleate giant cells of the HuT11 line also have been successfully established as continuous cell lines. The cloned lines have been morphologically distinct and more homogeneous, although typical giant cells have consistently appeared throughout the long-term culture of each. The HuT11 lines have grown as monolayers in McCoy's Medium 5A supplemented with 10% fetal calf serum, with generation times of 12 to 14 hr and high saturation densities. Cytogenetic studies showed that early and later passages of HuT11 cells were aneuploid, and all cell lines were successfully heterotransplanted in the hamster cheek pouch. Repeated indirect immunofluorescence examinations have shown each cell line to be negative for Epstein-Barr virus nuclear antigen. Indirect immunofluorescence tests in which monospecific immunoglobulins were used revealed positive membrane reactions for the gamma (heavy)-chain and kappa (light)-chain of human immunoglobulin G in approximately 20% of viable cells in each line; however, direct immunofluorescence with anti-human immunoglobulin G F(ab')2 reagent failed to confirm these reactions. Rosette tests for B- and T-lymphocyte and macrophage membrane receptors yielded negative results. All cell lines were strongly phagocytic for latex particles and neutral red dye. Cytochemical stains of the monolayers revealed abundant esterase, fluoride-resistant nonspecific esterase, acid phosphatase, and leucine aminopeptidase activities, while lysozyme assays were negative. Although some properties of the HuT11 lines have suggested a macrophage derivation, an undifferentiated lymphoid cell origin of the Hodgkin's neoplastic cell remains a possibility.