Fast, rate-sensitive corticosteroid negative feedback during stress. Characteristics of the fast, rate-sensitive, negative-feedback regulation of adrenocorticotropin secretion during stress was quantitatively analyzed using rats anesthetized with pentobarbital sodium. Various levels of plasma corticosterone were achieved during morning hours by infusing corticosterone solutions of different concentrations. Blood was sampled serially from the carotid artery. An increase in plasma corticosterone concentration 15 min after intravenous, pulsed injection of histamine (230 microgram) during saline intravenous infusion was defined as the "control response". When plasma corticosterone was rising during corticosterone infusion, the response to histamine stimulus was distinctly inhibited (fast, rate-sensitive feedback inhibition), whereas such an inhibition was not observed when plasma corticosterone levels were not rising, regardless of the absolute level. The critical rate of rise of plasma corticosterone, at or above which the fast rate-sensitive feedback was manifested, was 4-6 microgram/100 ml per min. When three graded doses of histamine were injected while plasma corticosterone levels were increasing at a rate of 6 microgram/100 ml per min, the absolute value of the inhibition observed was independent of the administered dose of the stressor. A hypothetical model for the mechanism of this feedback inhibition, based on the assumption that the hormone effect was proportional to the rate of formation of hormone-receptor complex, satisfied the quantitative characteristics of the inhibition experimentally observed in this study.


Angiotensin II infusion increases vasopressin, ACTH, and 11-hydroxycorticosteroid secretion. The effects of intravenous infusion of Asp1. Ile5-angiotensin II on blood pressure, plasma vasopressin, ACTH and 11-hydroxycorticosteroid levels and on plasma renin activity were studied in five trained, conscious dogs. The dogs were prepared with bilateral carotid loops. Infusion of angiotensin II at rates of 5, 10, and 20 ng/kg.min raised its plasma concentration from 23 +/- 7 to 48 +/- 8, 125 +/- 8, and 187 +/- 21 pg/ml, respectively. The lowest rate of infusion was mildly pressor, the two higher rates more so. All rates of infusion promptly increased vasopressin levels and depressed renin levels. The two higher rates also stimulated ACTH, although with a latency of 30-45 min. Since the rates of infusion of angiotensin II employed produced plasma levels within the physiological range, it is suggested that peripherally generated angiotensin II may play an important role in the regulation of vasopressin, and ACTH secretion.


A clinicopathologic study of steroid-related liver tumors. A registry of liver tumors was started in late 1973 in an attempt to assess the relationship of these tumors to oral contraceptives or to other environmental factors. This report is concerned with the pathological aspects and the possible pathogenesis of the first 101 tumors accessioned. There were 44 instances of focal nodular hyperplasia, 40 adenomas, four unclassified but probably benign tumors, and 13 hepatocellular carcinomas. Eighty-one patients took oral contraceptives; six were associated with pregnancy; three had taken estrogens for long periods of time; one had a thecoma; four never took sex steroids; and in five the history was unknown. Tumor rupture and intrahepatic hemorrhage were frequent complications. It is possible that the vascular lesions associated with focal nodular hyperplasia could play a part in their pathogenesis as well as with rupture. Foci of adenomatous hyperplasia may be related to the development of adenomas. The association of these tumors with sex steroids could be coincidental. The fact that none of the patients had cirrhosis of liver fibrosis, and that androgenic anabolic steroid therapy has been associated with hepatocellular carcinomas in males, suggests that further study of the problem is necessary.


Osteosarcoma of bone and its important recognizable varieties. Osteosarcoma of bone is a recognizable entity if the histopathologist designates tumors as such when their malignatn cells produce osteoid substance even if only in small foci. Such definition distinguishes this lesion from other sarcomas that arise in bone, especially chondrosarcoma and fibrosarcoma. There is a general tendency to consider that osteosarcomas represent a stereotyped form of disease for which new modalities of treatment can be applied and assessed. The question of whether a given osseous lesion is actually malignant and not a benign neoplasm or even a reactive non-neoplastic condition simulating a malignant tumor may be difficult for the histopathologist. Pathologists without considerable experience in the diagnosis of bone tumors find this question especially vexing. The establishment of a valid diagnosis of osteosarcoma introduces the additional problem that the 11 varieties considered in this paper may pose significant recognizable variations in the clinical capability of the disease. It is apparent that the physician must recognize the known clinicopathologic and prognostic factors of these subtypes in his assessment of the overall problem.


Pulmonary alveolar cell carcinoma. Fine structural and in vitro study of a case and critical review of this entity. Alveolar cell carcinoma is an unusual pulmonary neoplasm composed of cells with the ultrastructural features of granular pneumocytes. In the past this lesion has been grouped with peripheral well-differentiated mucin-secreting adenocarcinomas under the heading of bronchiolo-alveolar cell carcinoma. Because the existence of alveolar cell carcinoma as a distinct entity has been disputed, we examined a case ultrastructurally and confirmed that the neoplastic cells exhibit granular pneumocyte differentiation (lamellar bodies and abundant surface microvilli). In addition, we have growth this tumor in organ culture for up to 5 months and have documented the persistence of granular pneumocyte differentiation during this period. The accumulated evidence from human and animal studies that the granular pneumocyte may undergo neoplastic transformation is briefly reviewed. We conclude that alveolar cell carcinoma is a distinct entity; however, the elucidation of its natural history must await series that separate this lesion from other pulmonary carcinomas.


[The determination of age and sex from measure of the humerus (author's transl)]. On a total of seventy left humeri of persons in their third to ninth decades of life thirty different bone dimensions were determined macroscoically and checked for age dependence. Strong dependence on age was shown only by the weight and volume of the humerus as well as by the areal and thickness dimensins of the center of the shaft. These were used to set up multidimensional linear regression functions by which it is possible for the age of persons past thirty to be estimated with defined accuracy. Also calculated were discriminant functions which allow the sex of a person to be identified to be determined from three dimensions of the left humeral head, and these are the size of the caput at the cartilage-to-bone interface, the transverse and vertical diameters of the caput humeri or head of the humerus. Of the 49 cases studied only three were found to be in the region of overlap of the dimensions used for calculation. Consequently, the formula would not allow unknown humeri or skeletons, respectively, to be identified as belonging to either male or female persons in 6 percent of the cases only. The results of additional examinations made of 198 right humeri were less accurate and showed a decrease of the sexual dimorphism of bone with age.


[Determination of free crystalline silica in atmospheric dust]. The method for the determination of free crystalline silica (quarts), as previously described by two of the authors, has been employed on atmosphere dust of unconfined spaces. The matrix effect of atmospheric dust has been investigated in winter and in summer. The limits in the use of this method have been studied considering also what stated by law code n. 615. The results obtained on the outdoor atmospheric dust, samples from industrial as well as suburban areas, are reported. The results are evaluated in relation to the metereological records.


Humoral and cellular immune response during the growth of an SV40 induced tumour in hamsters. Hamster fibroblasts transformed in vivo by the SV40 virus (TSV5Cl2) induce tumours when injected into adult hamsters and antibodies present in sera of tumour-bearing animals are used to demonstrate the presence of the antigens specific for the viral transformation. These tumours are a very useful tool in studying the evolution of the immune response during the tumour growth. A systematic histological examination of the spleen, thymus and lymph nodes was undertaken and the results thus obtained were correlated with a parallel study of the cells of the peritumoral region, the thymus and the spleen by immunofluorescence using antisera of different specificities. We were able to show that the plasma cells which make up the early peritumoral reaction and the lymphoblasts found later in spleen and thymus both synthesize antibodies directed against virus induced antigens. Circulating antibody directed against the same antigens is first detected at/or about the time that the peritumoral plasma cell reaction disappears and increases progressively as tumour weight increases.


Immunoprophylaxis of experimental cytomegalovirus infection. Experimental MCMV infection simulates human disease. Since human CMV has no experimental animal, the murine model is useful to study mechanism of immunity to CMV and immunoprophylaxis of CMV infection. The live attenuated MCMV, obtained by serial passages in MEF, was effective in immunization of mice against fatal MCMV infection. Every parameter of the MCMV infection was nullified by the prior immunization. The protective immunity was cell-mediated and independent of antibody production. The sensitized T lymphocyte interfered with virus replication in vitro and in vivo. Efficacy and possible risk of a human CMV vaccine were discussed with the murine model.


Factors concerning the immunity to poxvirus infection. Two kinds of early antigens were shown in vaccinia-infected cells, one heat (56 degrees, 30 min)-stable (ES antigen) and the other heat-labile (EL antigen). Both antigens do not induce circulating neutralizing antibody in rabbits, but resistance to intradermal inoculation of active virus was observed when animals were immunized with either antigen in combination with Freund's complete adjuvant. Vaccinia-specific haemagglutinin (VHA) associated with infected cell membrane was purified and its antigenicity was also studied. Rabbits showed a good antibody response to VHA and some animals also showed neutralizing antibody, though in a small amount, to either intracellular (ICV) or extracellular (EVC) virus. Rabbits immunized with membrane antigens obtained from either VHA-positive or negative virus in combination with Freund's adjuvant showed skin restance to challenge with active virus. It was suggested that antibody against ECV might be responsible for the resistance.


Skeletal maturity of the hand and wrist in Japanese children in Sapporo by the TW2 method. Skeletal maturity of Japanese children aged 6--18 years in Sapporo was compared with that of Japanese children in Toko, as well as with that of British children, by the TW2 method. In Sapporo children, the mean skeletal age was in advance of the chronological age after ages 12 years in boys and 10 years in girls. The ages at which the skeletal age preceded the chronological age occurred earlier in the children in Tokyo than in Sapporo. The hand and wrist bones completed maturation about the same age in both Japanese and British children. Somatometric and radiological results agreed exactly.


A new placental protein test for the presence and identification of trophoblastic tumors. Trophoblast-specific beta1-glycoprotein (TSG) is detected in serum of patients with trophoblastic and nontrophoblastic tumors by means of immunodiffusion in agar (IDF), immunoradioautography in agar (IRA), and double-antibody radioimmunoassay (RIA), The identification of TSG is carried out by monospecific rabbit antisera. By IRA, TSG was found in serum of patients with trophoblastic tumors both before treatment (82.4%) and after surgery and/or chemotherapy (37.2%). All so-called falsepositives were associated with teratocarcinoma of the testicle. By means of RIA, TSG was also discovered in serum of patients with a variety of malignancies. In total, the TSG level was elevated in 15% of 114 cases of nontrophoblastic malignancies; as a rule, from 3 to 12 ng/ml. TSG was not detected in normal adult blood sera and in different pathological conditions.


[Quantitative amino acid makeup and the characteristics of the amino groups of ristomycin and the products of its partial acid hydrolysis]. The quantitative amino acid composition of ristomycin A, a glycopeptide antibiotic, peptides I-IV (from partial acid hydrolysis of the antibiotic) and their dinitrophenylic derivatives was determined. It was shown that both ristomycin and free peptides I-IV contained one residue of ristomycinic acid and one residue of actinoidinic acid, diamino-dicarbonic amino acids of the glycylphenolic type. Peptides I-IV had close molecular weights, i.e. 1100-1200 and differed from each other in the gradually increasing numbers of NH2- and COON- groups, from one in peptide I to four in peptide IV. The quantitative amino acid analysis of the dinitrophenylic derivatives of ristomycin and peptides I-IV showed that the free NH2-group in peptide I belonged to ristomycinic acid, the same as in the antibiotic, while in peptides III-IV at least one of the free NH2-groups belonged to ristomycinic acid and the other belonged to actinoidinic acid.


[Experimental study of the antiviral properties of antibiotic 6734-21]. The effect of antibiotic 6734-21 on the viruses of variolovaccine, Herpes simplex, influenza and classical avian plague was studied on various experimental models. Antibiotic 6734-21 inhibited development of the variolovaccine virus in the tissue culture, in chick embryos, in rabbits with variolovaccine infection, as well as the development of the viruses of Herpes simplex, Aueski, and Newcastle diseases in the tissue culture. It had a virulicidic effect on the viruses of variolovaccine, influenza and classical avian plague.


Large-scale production and concentration of infectious Epstein-Barr virus. Infectious Epstein-Barr virus (EBV) was produced from suspension cultures of P3HR-1 cells. A protocol for the scaled-up production and concentration of the virus was developed. Virus from the culture fluid was concentrated by continuous-flow pelletization or continuous flow with banding in sucrose. EBV prepared by pelletization yielded 1.7 X 10(4) infectious units/ml (100 X concentration) and less than 3.4 X  10(7) EVB particles/ml (1,000 X concentration), whereas EBV prepared by banding yielded 4.6 X 10(3) infectious units/ml (100 X concentration) aand 1.3 X 10(8) EBV particles 1 ml (1,000 X concentration). The majority of the virus particles observed were "empty" membrane-associated particles. A statistical test of the correlation between infectivity and virus particle count was made.


Minimal breast cancer: a clinical appraisal. Eighty-five patients with a diagnosis of minimal breast cancer were evaluated. The predominant lesion was intraductal carcinoma, and axillary metastases occurred in association with minimal breast cancer in seven of 96 cases. One death occurred due to minimal breast cancer. Bilateral mammary carcinoma was evident in 24% and bilateral minimal breast cancer in 13% of the patients. The component lesions of minimal breast cancer have varied biologic activity, but prognosis is good with a variety of operations. The multifocal nature of minimal breast cancer and the potential for metastases should be recognized. Therapy should include removal of the entire mammary parenchyma and low axillary nodes. The high incidence of bilateral malignancy supports elective contralateral biopsy at the time of therapy for minimal breast cancer.


Evaluation of cardiac ischemia by NADH fluroescence photography. A direct, noninvasive method of assessing the oxidation-reduction potential of an intramyocardial respiratory chain component is described. The technique is based on the differences in spectral properties between the oxidized and reduced forms of nicotinamide adenine dinucleotide (NADH). The tissue surface fluorescence from intracellular NADH may be measured and documented photographically. Noose occlusion of a coronary artery produced detectable NADH fluorescence in 15 seconds in the subtended ischemic epicardium. This fluorescence of reduced pyridine nucleotide resolved following 60 seconds of reperfusion of the ischemic myocardium. The reduction of epicardial NADH with ischemia is a rapid and reversible process. A subsequent noose reocclusion resulted in a reproducible pattern of fluorescence. The technique of NADH fluorescence photography appears superior to current methods of assessing tissue oxygen supply:demand.


The beta-adrenergic activity of some monosubstituted phenethanolamines. A series of monosubstituted phenethanolamines with N-isopropyl and N-tert-butyl substituents has been prepared. A detailed pharmacological study has been made of the beta-adrenergic activity of these materials as the nitrogen substituent and the nature and position of the phenyl substituent were changed, and their selectivity has been determined for beta1- and beta2-adrenoceptors in guinea-pig and cat tissues.


A comparative study of the beta-adrenoceptors of guinea-pig, rabbit and rat trachea. The sensitivity to isoprenaline of beta inhibitory adrenoceptors of tracheal chain preparations from guinea-pig, rabbit and rat was compared. For this purpose the tracheal chain preparations were contracted either with KCl or pilocarpine. Isoprenaline (1.0 X 10(-7)M--6.4 X 10(-6)M) caused concentration-related relaxation of guinea-pig trachea, whereas the rabbit and the rat trachea remained insensitive even at very high concentrations of isoprenaline (2.5 X 10(-3)M). Phenoxybenzamine (2.6 X 10(-6)M) potentiated or unmasked the relaxant effects of isoprenaline respectively in the guinea-pig or the rabbit and the rat trachea contracted with KCl. Phentolamine was ineffective in potentiating or unmasking the relaxant effects of isoprenaline. The potentiating or unmasking effects of phenoxybenzamine were absent in reserpinized preparations and lidocaine treated preparations from both guinea-pig and rat. Normetanephrine (5.4 X 10(-7)M) potentiated responses to isoprenaline only in the guinea-pig trachea. Even a 10-fold higher concentration of normetanephrine was ineffective in the rabbit or the rat rachea.


The influence of light and different ATP concentrations on cell aggregation in cyclic AMP sensitive and insensitive species of the cellular slime molds. The influence of light and different concentrations of ATP on cell aggregation in cyclic AMP sensitive (Dictyostelium mucoroides, D. purpureum) and cyclic AMP insensitive species (Polysphondylium violaceum, P. pallidum, D. lacteum) of the cellular slime molds was observed in small and in large amoebal populations. Both light and ATP (optimal concentration: 10(-5) M) accelerated cell aggregation and increased the number of aggregating centers in large populations. For cyclic AMP sensitive species the effect of ATP in large populations was more pronounced than for the species that do not react to cyclic AMP. A possible explanation for the similar effect of light and ATP has been discussed.


Metabolic regulation in Pseudomonas oxalaticus OX1. Enzyme and coenzyme concentration changes during substrate transition experiments. Metabolic control associated with diauxic growth of Pseudomonas oxalaticus in batch cultures on mixtures of formate and oxalate was investigated by measuring intracellular enzyme and coenzyme concentrations and QO2 values during transition experiments from oxalate to formate and vice versa. In transition from oxalate to formate oxalyl-CoA reductase concentration declined after the exhaustion of oxalate and ribulose-1,5-diphosphate carboxylase and 14CO2 fixation appeared upon addition of formate. In the reciprocal transition, ribulose-1,5-diphosphate carboxylase and 14CO2 fixation rate declined sharply after formate exhaustion, and oxalyl-CoA reductase appeared only after addition of oxalate. The intracellular NAD and NADP concentrations measured in the same experiments are reported. At substrate exhaustion the proportion of NAD in the reduced form fell from 15-20% to 2%. On addition of formate to an oxalate-starved culture there was an immediate increase in the proportion of NADH to 50%; such an increase was not observed in the reverse experiment.


The alternate respiratory pathway of Candida albicans. Candida albicans contains a cryptic cyanide and antimycin A insensitive respiratory system. This alternate oxidase was found (i) at all growth rates from mu = 0.05 to 0.26 in a chemostat culture and (ii) in both mycelial and yeast forms of the organism. Neither chloramphenicol nor cycloheximide prevented the expression of the alternate oxidase. Salicylhydroxamic acid was a potent inhibitor of the cyanide insensitive respiration. The respiration of mitochondria grown in the presence of antimycin A was not inhibited by cyanide or antimycin A but was inhibited by salicylhydroxamic acid.


[Causes for species difference in acute toxicity of chlorocholine chloride]. Chlorocholine chloride (CCC) inhibits neuromuscular transduction of excitation and, consequently, leads to respiratory arrest in cases of acute intoxication. An account is given of the relationships between neuromuscularly blocking activity and acute toxicity of CCC. Several animal species and pharmacological models are used to produce evidence to the effect that CCC-caused inhibition of neuromuscular transmission of excitation is characterised by parameters typical of block due to depolarisation. The differentiated sensitivity of species to depolarising neuromuscular blockers is thought to be the decisive cause of species differences regarding acute toxicity of CCC. Conclusions are discussed which may be derived from the above findings regarding acute CCC toxicity to man and agricultural animal.


[Experimental infection of man using viral strains of bovine papular stomatitis, orf, pseudocowpox and milker's nodule]. An account is given of the close correlations that exist between virus strains of bovine papular stomatitis, orf, pseudocowpox, and milker's nodule. Reference is made to literature data on natural infection of man with the above virus strains. A report then is presented on experimental infection of human volunteers, using paravaccine birus. While fairly tough and elevated nodules, 4 mm to 5 mm in diameter, were produced on the probends' skin, no re-isolation of virus was achieved.


Inorganic cytoplasmic inclusions in alveolar macrophages. The role of cigarette smoking. A case of tobacco-associated pulmonary fibrosis, with the results of histological, ultrastructural, and spectrometric analysis is reported. Abnormalities of the alveolar macrophages, which are particularly affected by tobacco inhalation were found. The size of the macrophages was increased and many large, polymorphous inclusions, including fat vacuoles and granular deposits, which were either homogeneous or electron lucent vacuoles, were seen in the cytoplasm. A few laminar structures were observed. All of these lesions are frequently found in cigarette smokers. Still more interesting was the discovery of numerous fiber-, needle-, or laminar-like inclusions that varied in size from 0.2 to more than 2 mu. The digestions of the inclusions with potassium hydroxide confirmed the presence of various metals, such as sodium, magnesium, potassium, iron, sulfur, and especially, aluminum, and silicon; these last two elements correspond to the presence of kaolinite in the tissue, as has been previously described, and can be considered as evidence of the use of tobacco.


Malignant thymic tumor in an infant (malignant histiocytoma). A 2-month-old infant was diagnosed as having an invasive thymic tumor. Histologically and ultrastructurally, the tumor was composed of primitive mesenchymal cells, most likely malignant histiocytes. The child died at the age of 5 1/2 months with metastases to the bone and and liver. Review of the literature on thymic tumors and fibrous histiocytomas or malignant histiocytoma shows no similar case.


Phosphatase enzymes. Cytochemical study of pleomorphic adenoma and normal human salivary glands. Human parotid glands, submandibular glands, and pleomorphic adenomas were examined by electron microscopic histochemistry. All epithelial cells of the normal salivary glands showed plasma membrane adenosine triphosphatase (ATPase) and inosine diphosphatase (IDPase) activity. However, myoepithelial cells reacted most intensely. Pleomorphic adenomas showed epithelial cells within solid and ductal portions of the tumors that were variably reactive for both ATPase and IDPase. Histochemical examination of the epithelial cells in the myxoid portions of the tumors did not provide conclusive evidence as to the nature of their progenitor cells. Surface-associated phosphatases (alkaline phosphatase, ATPase, and IDPase) cannot be reliably used as histochemical markers of salivary gland myoepithelial cells. Therefore, morphological and phosphatase histochemical studies that intend to examine the role of myoepithelial cells in salivary gland neoplasms must be interpreted with care.


Giant cell tumors of tendon sheath. An electron microscopical study of 11 cases. Light and electron microscopy of 11 giant cell tumors of tendon sheath revealed a pleomorphic cell population in which the giant cells had similarities to osteoclasts, and the stromal cells had similarities to primitive mesenchymal cells, osteoblasts, fibroblasts, and histiocytes. I suggest that giant cell tumors of tendon sheath are derived from mesenchymal cells with partial osseous differentiation.


[Metastasis of breast cancer (according to findings of the pathologist of the I. M. Sechenov Moscow Medical Institute I during the 25 year period -- 1951-1975)]. The mammary gland cancer metastasizes most frequently into lymph nodes, the liver, bones, lungs and adrenal glands. Calculations of the combined coefficient of metastasizing showed the adenocarcinoma more frequently than other forms to metastasize into the lymph nodes of the chest, adrenals, and kidneys and less frequently into the liver; solid carcinoma with pseudoepidermoid metaplasia metastasizes into the brain more frequently than the other forms. The longest average survival after treatment (radical mastectomy, radiation therapy) was established in a more differentiated form of cancer - adenocarcinoma; the survival time after solid carcinoma and fibrous cancer is considerably shorter.


Corpora amylacea of the lumbar spinal cord and peripheral nervous system. Tissue from L-4 spinal cord, dorsal root ganglia, sural nerve, and intramuscular branches to the gastrocnemius was obtained during 35 random autopsies, embedded in paraffin, and stained with PAS and Holmes Alcian blue and studied for the incidence and distribution of corpora amylacea (CA). Intraaxonal CA in spinal grey matter were commonly found, but the incidence in root ganglia, sural, and intramuscular nerve was low. Clinically insignificant intraneuronal spinal grey CA were found in six of eight men past the age of 60 years. Corpora amylacea in spinal white matter were most common in the region of posterior root entry.


Herpes simplex virus types 1 and 2 in ocular disease. In a continuous series of 457 patients with presumed herpetic eye disease, virus isolation and typing revealed 154 patients with herpes simplex virus type 1 and three patients with type 2 infections. In 219 isolates that were examined for neutralization by specific antisera, for growth in human fibroblasts, and, in part, for temperature sensitivity, there were found substantial strain differences in addition to the type-specific characteristics. The clinical features of each of the three type 2 infections are described in detail. A suggestive correlation found between clinical courses and virus growth characteristics of the type 1 strains indicates that further virologic differentiation of these strains would be useful.


Iris metastasis from a bronchial carcinoid tumor. A 54-year-old man had a fleshy pink vascularized mass in the superonasal quadrant of the right iris. He had a history of an unchanging pulmonary nodule that was followed up for five years with routine chest roentgenograms. Fluorescein angiography demonstrated early filling of numerous fine blood vessels in the mass and diffuse late staining of the lesion. Results of a transcorneal radioactive phosphorus (32P) uptake test were positive. The clinical diagnosis was tapioca melanoma of the iris, with metastatic tumor as a second possibility. The tumor was excised by iridocyclectomy. Light microscopy revealed nests of tumor cells with oval hyperchromatic nuclei. Fontana stains for melanin were negative as were argentaffin and argyrophil strans. Electron microscopy demonstrated light and dark tumor cells containing numerous membrane-bound electron-dense neurosecretory granules. The diagnosis was iris metastasis from a probable bronchial carcinoid. The patient is alive and well years after excision of the iris mass.


[Pathophysiology and metabolism in osteogenesis imperfecta (author's transl)]. The tissue changes in osteogenesis imperfecta apparently lie in the collagen and in the glycosaminoglycans. The collagen shows structural development disturbance and incomplete calcification. The tissue glycosaminoglycans are increased. The basis probably lies in a disturbed ATP metabolism (Solomons and Millar, 1973). An increased pyrophosphate concentration explains the decreased calcification and the structural changes. The increased osteolysis could be explained by increased quantities of cAMP.


[Studies of the mucopolysaccharid metabolism in the juvenile angiofibroma (author's transl)]. The mucopolysaccharide content of juvenile angiofibromas is investigated using histochemical and ultrahistochemical methods (histochemical in ten cases, ultrahistochemical in four cases). The investigations result in typical histochemical features and show the localization of acid mucopolysaccharides at electronmicroscopical level. The endothelium of the vessels exhibits a strong PAS-reaction and also shows acid groups at the cell surfaces. The interstitial tissue demonstrates neutral perjodatreactive substances as well as acid mucopolysaccharides. The latter substances mainly reacted in perifibrillar position. Electronmicroscopically the perjodreactive material was localized in the cytoplasma of typical fibroblasts and their modulations resembling histiocytes. The acid groups of carboxylated and sulfated mucopolysaccharides of the intercellular substance could be determined as hyaluronic acides and chondroitinsulfates (Chondroitin-4 and Chondroitin-6 sulfate) using the critical-electrolyte-concentration method after Scott and Dorling. Ultrastructurally these acid groups are localized as cell coat covering the fibroblasts and between the collagen fibrils. This histochemical investigation also supports the thesis of an organoid growth pattern of juvenile angiofibroma.


Changes in ornithine decarboxylase activity and cyclic adenosine-3'-5'-monophosphate concentrations during the cell cycle of synchronized BHK cells. BHK cells were synchronized by excess thymidine treatment, which resulted in approximately 90% synchrony. The activity of ornithine decarboxylase (ODC), the rate-limiting enzyme in polyamine biosynthesis, elevated in early S phase, decreased in G2 + M and G1 phase and then increased during late G1 approximately second round of early S phase. The concentration of cyclic adenosine-3'-5'-monophosphate (cAMP) gradually decreased during S approximately G2 + M phase and then increased during late G1 approximately second round of early S phase, preceding that of ODC activity. The data suggest that ODC activity might be regulated by cellular cAMP level.


Requirements for stimulation of T cell responses against virus-infected cells: nature of ectromelia virus-infected cells capable of stimulating cytotoxic T cells in a secondary response in vitro. The nature of infected stimulator cells in the in vitro secondary cytotoxic T cell response to ectromelia infection was investigated. It was found that macrophages were better stimulator cells than spleen cells. B cells (Ig-positive cells) were superior to T cells (Ig-negative cells) both on a relative proportion and on a cell-to-cell basis. Concanavalin A and lipopolysaccharide-stimulated lymphocytes were also effective stimulator cells but appeared to be slightly inferior to spleen cells. Spleen cells depleted of Ia-positive cells were markedly inferior to normal spleen cells as stimulators. It was also found that primary and secondary cytotoxic T cells were largely Ia-negative. These findings are discussed in relation to the likely events during T cell responses to infection in vivo.


Investigations on the turnover of adrenocortical mitochondria. XI. Effects of dexamethasone on the half-life of mitochondria from the rat zona fasciculata. The effects of dexamethasone on the half-life of rat adrenocortical mitochondria were investigated by high resolution autoradiography and liquid scintillation spectroscopy. According to the method employed, the half-life averaged 11.27 and 10.46 days. Dexamethasone was found to decrease significantly this parameter (to about 5 days). Since dexamethasone-treated rats can be regarded as pharmacologically hypophysectomized animals, the data indicates that ACTH is involved in the maintenance of the half-life of adrenocortical mitochondria.


Possible mitochondrial involvement in mechanism of cytoplasmic male sterility in maize (Zea mays L.). The mechanism of cytoplasmic male sterility was investigated in maize by isolating mitochondria from seedlings and various anther stages and analyzing cytochrome oxidase and succinic dehydrogenase biochemically and electrophoretically. Sterile anthers exhibited a lack of biochemical activity and fewer isozymatic bands for cytochrome oxidase. No apparent differences were detected biochemically or electrophoretically between fertile and sterile anthers for succinate dehydrogenase.


Regulation of the neutral phosphatase in Chlamydomonas reinhardi: an immunogenetic study of wild-type and mutant strains. In Chlamydomonas reinhardi, the activity of the neutral phosphatase considerably increases when the cells are grown in the absence of inorganic phosphate (Pi). A comparative immunological study of cells grown on media containing Pi or not indicated that the neutral phosphatase was synthesized de novo. Ten mutants lacking the neutral phosphatase and distributed among three genetic loci (PD2, PD3, PD24) were investigated for their ability to produce cross-reacting material (CRM) antigenically related to the wild enzyme. All mutants were shown to form much less CRM than the wild-type strain. It is proposed that the three genes are involved in the regulation of neutral phosphatase synthesis.


Hydrolysis of synthetic pyrophosphoric esters by an isoenzyme of apyrase from Solanum tuberosum. A highly purified isoenzyme of apyrase obtained from potatoes (Solanum tuberosum var. Pimpernel) exhibits a low specificity for the organic moiety of synthetic pyro- and triphosphates. Methyl di- and tri-phosphates were hydrolysed at higher rates than ADP and ATP, but their Km values were also higher. Steric hindrance at the carbon atom linked to the pyrophosphate chain decreases both binding and maximum rate, whereas length or polarity of the organic chain do not have systematic effects. t-Butyl diphosphate, inorganic pyrophosphate, adenosine 5'-[alpha,beta-methylene]triphosphate and adenosine 5'-[beta,gamma-methylene]triphosphate are competitive inhibitors of the hydrolysis of ATP and ADP.


Mixed-valence cytochrome oxidase-formate complex. A steady-state intermediate. At neutral pH, formate binds to the haem a3 component of cytochrome c oxidase to give a complex that reacts differently from the non-liganded enzyme with reducing agents. Addition of sodium dithionite to the formate complex leads directly to the formation of the fully reduced species, whereas reduction with ascorbate/tetramethylenephenylene-diamine can lead to the production of a mixed-valence species. The stability of this mixed-valence form was studied, and the species appears to represent a 'steady-state' situation that is stable only in the presence of an excess of O2 and reducing equivalents. Characterization of the mixed-valence complex by electron paramagnetic resonance and magnetic circular dichroism reveals the presence of reduced low-spin haem a together with reduced detectable copper and high-spin ferric haem a3.


Chemotherapeutically active nitro compounds. 4. 5-Nitroimidazoles (Part I). More than 135 new 2-methyl-5-nitroimidazoles substituted in 1-position and 1-methyl-5-nitroimidazoles substituted in 2-position were investigated for their activity against various protozoan species, in particular Entamoeba histolytica in the golden hamster, Trichomonas fetus, Trypanosoma brucei and T. cruzi in the NMRI mouse. Among the nitroimidazoles substituted in the 1-position only two preparations exhibited a similar effect as metronidazole preparations exhibited a similar effect as metronidazole against T. fetus. In the class of the nitroimidazoles substituted in the 2-position 16 compounds were as effective as metronidazole, 19 showed an effect superior to metronidazole, 1 was as good as tinidazole and 2 exhibited an activity superior to tinidazole against T. fetus. Only few compounds displayed any amoebicidal activity. Of the mono and bis-hydrazones of 1-methyl-5-nitroimidazole-2-aldehyde substituted in the 2-position 3 compounds had an amoebicidal effect 2 to 8 times stronger than that of metronidazole. Only few representatives of the 1-methyl-5-nitroimidazoles substituted in the 2-position produced a useful trypanocidal effect when given in relatively high doses. The structure-activity relationship of 5-nitroimidazole derivatives has been discussed.


[Diagnosis of Rotavirus using viral RNA electrophoresis]. Rotavirus has emerged as a major agent of acute enteritis of infants. A rapid and simple technique has been developed for its detection, based on the visualization of the viral RNA electrophoretic pattern. This technique takes advantage of the large amount of rotavirus present in the stools and the high sensitivity for detection of RNA obtained with ethidium bromide.


[Existence of 2 types of Rotavirus associated with acute gastroenteritis in children]. Rotaviruses found in the stools of different infants, hospitalized in the Hospital del Niño del DIF, may be grouped into at least two types according to their RNA composition. The viral RNA composition was analyzed by electrophoresis in agarose gels. The different types can be distinguished by the mobility of the second larger of the 11 segments that make up the viral RNA.


Some effects of the aminoglycoside antibiotic amikacin on neuromuscular and autonomic transmission. The effects of the new aminoglycoside antibiotic amikacin on neurohumoral transmission were tested in the anaesthetized cat, and in mouse, rat and chick isolated nerve-muscle preparations. Amikacin had blocking actions on both autonomic and neuromuscular transmission. The autonomic effects were caused mainly by ganglion blockade and were reversed by calcium. The amikacin-induced neuromuscular blockade reculted from a decreased release of acetylcholine and a reduced postjunctional sensitivity. Intracellular recording from end-plates in the rat diaphragm demonstrated that amikacin had magnesium-like effects on acetylcholine release. The blockade was reversed completely by calcium, 4-aminopyridine and 3,4-diaminopyridine and partially by neostigmine. The neuromuscular effects of amikacin in vivo were augmented greatly after pretreatment with tubocurarine. It is concluded that care should be exercised if amikacin is administered during surgery in conjunction with tubocurarine.


Magnitude, dose-requirement and mode of development of tachyphylaxis to suxamethonium in man. Tachyphylaxis to suxamethonium infused at a constant rate was studied in 15 surgical patients under enflurane-nitrous oxide-oxygen anaesthesia. The compound electromyographic response of the adductor pollicis muscle to stimulation of the ulnar nerve was monitored. A short-lasting and a long-lasting nearly steady state were observed in Phase I and in Phase II, respectively, during both of which a constant infusion resulted in a constant block. Tachyphylaxis occurred during the transition of phases, beginning after infusion of 1.4 +/- 0.6 (SD) mg kg-1 of suxamethonium, and 36 +/- 14 (SD) minutes of exposure. Tachyphylaxis peaked after 2.6 +/- 1.3 (SD) mg kg-1, and 72 +/- 38 (SD) minutes of exposure. The maximum gain in neuromuscular transmission from the initial maintenance level of block as the result of tachyphylaxis was 53 +/- 24 (SD) % of the control. The results are thought to reconcile previously reported conflicting clinical observations on tachyphylaxis.


Attempt to demonstrate an in vivo effect of mianserin hydrochloride on erythrocyte Na+-K+-ATPase activity and cyclic AMP concentration. 1. There is evidence that erythrocyte Na+-K+-ATPase activity and erythrocyte cyclic AMP change on recovery from a depressive illness. Mianserin is a recently introduced antidepressant but its mode of action is unknown. The present study was therefore designed to investigate in vivo the effect of mianserin on erythrocyte Na+-K+-ATPase and cyclic AMP. 2. Biochemical estimations were made on blood from depressed patients before beginning either mianserin or matched placebo treatment, after 1 week, and again after 2 weeks' treatment. 3. Neither the erythrocyte Na+-K+-ATPase, nor the erythrocyte cyclic AMP concentration, changed significantly in either the mianserin- or the placebo-treated group. 4. The study sheds no light on the possible mechanism of action of mianserin.


Rheumatoid factor in sera of dermatitis herpetiformis patients. Serum specimens from 44 patients with dermatitis herpetiformis (DH) and 42 control patients were analyzed with solid-phase radioimmunoassay (RIA) developed to detect IgG and IgM class antibodies against herpes simplex virus (HSV) envelope antigen. They were also tested with different tests to detect rheumatoid factor (RF) as well as serum antibodies to gastric parietal cells, glomerular basement membrane, smooth muscle, mitochondria and nuclei. Total serum IgG, IgM and IgA quantification was performed concurrently. IgM class antibodies reacting in HSV envelope antigen RIA were found in 47.8% of the DH patients, but in none of the controls. The age distribution of the IgM positive patients was in accordance with the appearance of RF, representing elderly persons. The presence of RF in serum specimens was confirmed by removing the IgM reactivity with heat aggregated human gamma globulin adsorption and in 13 patients with positive serological tests for RF. No correlation was found with the appearance of RF and duration or severity of the disease or with the detection of RF and serum total IgM concentration. In DH patients antibodies to gastric parietal cells were found in 6 and to smooth muscle in 1 of 32 specimens tested.


Mechanisms of microtubule diassembly in vivo: studies in normal and chronic granulomatous disease leucocytes. Microtubule assembly in human polmorphonuclear leucocytes is a dynamic process that can be initiated by binding of the plant lectin Concanavalin A to surface receptors. Colchicine inhibits lectin-induced microtubule assembly and promotes the movement of Concanavalin A into surface caps. Inhibition of microtubule assembly and enhanced Concanavalin A cap formation also follow treatment of normal leucocytes with two specific glutathione-oxidizing agents, 'diamide' and tertiary butylhydroperoxide. Our objective here was to determine if microtubule inhibition is mediated via glutathione disulphide or via hydrogen peroxide that is generated in Concanavalin A-treated leucocytes and may accumulate when cells are depleted of reduced glutathione. We show that exogenous hydrogen peroxide induces Concanavalin A capping on normal polymorphonuclear leucocytes but only at concentrations that simultaneously oxidize glutathione. We also show that 'diamide' and tertiary butylhydroperoxide cause Concanavalin A cap formation in leucocytes from patients with chronic granulomatous disease. These cells cannot generate significant amounts of superoxide or hydrogen peroxide. Thus, it seems likely that the reversible inhibition of microtubule assembly and function caused by glutathione oxidants requires only increased glutathione disulphide and is not dependent on subsequent accumulation of other metabolites.


Impaired neutrophil function and myeloperoxidase deficiency in myeloid metaplasia. The ability of neutrophils to phagocytose and kill Candida guilliermondii was investigated in 12 patients with myeloid metaplasia (MM). Following ingestion there was a considerable impairment in the ability of MM neutrophils to kill and digest Candida which was not explained by the very mild impairment in phagocytosis. Quantitative myeloperoxidase measurement revealed an overall deficiency of this enzyme in MM neutrophils and a highly significant correlation between low myeloperoxidase levels and impaired candidacidal activity. Neutrophils from patients with myeloid metaplasia show a pattern of defective microbial killing, high alkaline phosphatase activity and low myeloperoxidase activty which is similar to that seen in severe infections and distinct from chronic granulocytic leukaemia. The cells of one patient with particularly low myeloperoxidase and defective microbial killing were further studied both cytochemically and by electron microscopy. The azurophilic granules of his neutrophils were present in normal numbers and contained normal amounts of acid phosphatase but they lacked myeloperoxidase.


Association of Pseudomonas cytochrome oxidase with liposomes. Purified Pseudomonas cytochrome oxidase has been associated with asolectin liposomes by two different methods. Firstly, the enzyme was attached to liposomic membranes by adding it to a cholate-phospholipid dispersion and subsequently dialyzing the detergent out of suspension. In the second case the enzyme was adsorbed on the preformed liposomes when added to them after the dialysis. A stimulation of the cytochrome oxidase activity approximately twenty-fold was observed by the first method. In contrast, the activation was absent in the second type of preparation, indicating that interaction between the enzyme and phospholipids is very different in the two types of vesicles. The cholate-dialysis method for reconstitution of protein-phospholipid vesicles seems to lead to rather heterogenous preparations. These can be further fractionated, not only according to their size but also to the protein/phospholipid ratio, by gel chromatography.


Phospholipid dependence of UDP-glucuronyltransferase. Very extensive hydrolysis of phospholipids with pure Bacillus cereus phospholipase C at 5 degrees C greatly inhibited the maximum demonstrable rate of glucuronidation of p-nitrophenol by UDPglucuronyltransferase in guinea pig liver microsomes. Lysophosphatidylcholine restored much of the inhibited activity but non-phospholipid surfactants or hydrolysis of diglycerides failed to reactivate. Phospholipid depletion likewise inhibited o-aminophenol glucuronidation and phospholipids restored activity. It is concluded that glucuronyltransferase specifically requires phospholipids for optimal activity. It seems unlikely that these phospholipids only serve to dissolve aglycones, or that they are direct physiological regulators of the transferase. Instead, a permissive role is ascribed to phospholipids, allowing glucuronyltransferase to be regulated by other means.


A model for ganglioside behaviour in cell membranes. Gangliosides from beef brain have been spin-labeled using two different attaching groups and employed to investigate the physical nature of ganglioside behaviour in membranes. Results obtained using EPR spectroscopy indicate that, in phosphatidylcholine bilayers at physiological pH, ganglioside oligosaccharide chains are quite mobile and show a measurable tendency towards cooperative interaction amongst themselves. We suggest that the source of this interaction is the formation of H-bonds between sugar residues in adjacent ganglioside molecules. We present evidence that physiological (extracellular fluid) levels of Ca2+ and Mg2+ lead to cross-linking and condensing of ganglioside headgroups by complexing sialic acid carboxyl residues. Ganglioside headgroup interactions are not very sensitive to changes in the buffer ionic strength, suggesting that ionic interactions are of minor importance. We have found no measurable tendency for headgroup carbohydrate to penetrate hydrophobic regions of lipid bilayers. EPR spectroscopy was also used to follow the interaction of spin-labeled gangliosides with the glycoprotein, glycophorin, and with intact BHK cells. We suggest that these carbohydrate-based interactions should contribute significantly to the properties of the eucaryotic cell glycocalyx. We predict that laterally mobile carbohydrate-bearing components of cell surface will show a tendency to cluster about complex glycoprotein arrays, especially if the species involved bear accessible carboxylic acid functions.


Regulation of de novo purine biosynthesis in normal and 8-azaguanine-resistant Chinese hamster cells. Azaguanine-resistant mutants of Chinese hamster ovary cells were isolated following mutagenesis with ICR-17OG. Of the eight mutant isolates examined, only one, Ag-5 had detectable hypoxanthine(guanine)phosphoribosyltransferase activity. Under normal conditions of growth, de novo purine biosynthesis in the mutants was not significantly different from wild type. However, when the cultures were starved for glutamine over a 2 h period before measuring 5'phosphoribosyl formylglycinamide (a relative measure of de novo purine biosynthesis), elevated levels of 5'-phosphoribosyl formylglycinamide accumulated in some of the mutants, and decreased levels in wild type and Ag-5. The level of purine biosynthesis could be related to the levels of glutamine in the pregrowth medium. The rate of purine biosynthesis correlated with 5-phosphoribosyl pyrophosphate levels, which were enhanced in the mutant (Ag-C) following the starvation period. No alterations were found in levels of 5-phosphoribosyl pyrophosphate synthetase or glutamine synthetase. The extent of feedback inhibition was normal in both mutant and wild type cells. These data suggest that the hypoxanthine (guanine) phosphoribosyltransferase locus is a regulatory gene.


Phosphorylation of calf thymus RNA polymerase II by nuclear cyclic 3',5'-AMP-independent protein kinase. Nucleoplasmic RNA polymerase II (nucleosidetriphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) from calfthymus is phosphorylated by homologous cyclic AMP-independent protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37). Polyacrylamide gel electrophoresis of the 32P-labeled RNA polymerase II under non-denaturing conditions revealed that both forms of the enzyme were phosphorylated. Polyacrylamide gel electrophoresis of the 32P-labeled RNA polymerase II under denaturing conditions showed that the 25 000 dalton subunit was the phosphate acceptor subunit. Partial acid hydrolysis of the 32P-labeled RNA polymerase II followed by ion-exchange chromatography revealed serine and threonine as the [32P]phosphate acceptor amino acids. Phosphorylation of the RNA polymerase II was accompanied by a stimulation of enzymatic activity and was dependent upon the presence of ATP.


Electrophoretic behaviour of yeast mitochondrial translation products. We have studied the mobility of yeast mitochondrial translation products during electrophoresis on polyacrylamide gels of different composition and found that these polypeptides can be divided into two groups. One, to which subunit II of cytochrome c oxidase belongs, behaves normal as all water-soluble reference proteins. The other, to which cytochrome b and subunits I and III of cytochrome c oxidase belong, shows a free electrophoretic mobility about twice as fast as the first group. Conditions have been found to separate cytochrome c1 from cytochrome b.


Inhibition of malate dehydrogenase by thyroxine and structurally related compounds. The inhibition of pig heart mitochondrial malate dehydrogenase (L-malate: NAD+ oxidoreductase, EC 1.1.1.37) by the thyroxine and structurally related compounds was studied to resolve a longstanding question about the exact nature of the inhibition. Thyroxine, in freshly prepared solution, was found to be a "pure" competitive inhibitor relative to the nucleotide cofactor. Upon standing in diffuse daylight, solutions of thyroxine showed increased ability to inhibit the enzyme, presumably as a result of oxidation of enzyme sulfhydryl groups by free iodine that is released photochemically. This behavior probably accounts for earlier reports of irreversible inactivation by thyroxine. Comment is made on the implications of these findings to the mechanism of thyroid hormmone action.


Increased activity of cyclic AMP phosphodiesterase from frozen-thawed rat liver. A role of lysosomal protease in enzyme activation. The activity of cyclic AMP phosphodiesterase (3':5'-cyclic-nucleotide 5'-nucleotidohydrolase, EC 3.1.4.17) in 105 000 X g supernatant fraction from frozen-thawed rat liver was 2.5 times higher than the corresponding preparation from fresh liver. This increased activity of frozen liver enzyme was accompanied by a decreased sensitivity of the enzyme to known activators such as alpha-tocopheryl phosphate and trypsin. Neither membrane-bound cyclic AMP phosphodiesterase, nor supernatant cyclic GMP phosphodiesterase increased in frozen liver preparation. It is unlikely that the activator protein of phosphodiesterase participated in the observed change of enzyme activity. Among rat tissues so far tested, the increased level of cyclic AMP phosphodiesterase was noted only in tissues rich in lysosome content. In the recombination experiment where phosphodiesterase from fresh liver was incubated with lysosomal fraction, stimulation of the enzyme activity was observed with a concomitant loss of sensitivity to above-mentioned activators. Since the stimulation by lysosomal fraction was effectively inhibited by cathepsin B1 inhibitors, leupeptin and antipain, it was deduced cathepsin-B1 (EC 3.4.12.3) type protease(s) was the main causative of activating the cyclic AMP phosphodiesterase. The freezing-thawing process of rat liver made the lysosomal membrane more permeable, and hence lysosomal proteases were released into soluble fraction during phosphodiesterase preparation. These results provide a warning not to use frozen liver for phosphodiesterase preparation, otherwise altered properties of the enzymes will be seen.


The catabolism of human and rat very low density lipoproteins by perfused rat hearts. The catabolism of human and rat 125I-labelled very low density lipoproteins (VLDL) was compared by perfusing the lipoproteins through beating rat hearts. Triacylglycerol was removed from the VLDL to a greater extent than the protein moiety, leaving remnants containing relatively more apo-B and less apo-C. The change in apo-C content of the remnants correlated with the loss of triacylglycerol. The extent of removal of triacylglycerol from the rat and human VLDL was similar and in most cases appeared to saturate the heart lipoprotein lipase. The remnants were slightly smaller in size than the VLDL, and included particles which appeared to be partially emptied. In addition to remnants of d less than 1.019 g/ml, iodinated lipoproteins derived from rat and human VLDL were recovered at d 1.019-1.063 and 1.063-1.21 g/ml. The former contained largely cholesterol and cholesteryl esters, while phospholipids were the dominant lipid in the latter. An average of 40% of the 125I-labelled apoprotein lost from the VLDL was associated with the perfused hearts. Very little d 1.019-1.063 g/ml lipoprotein was produced from low (physiological) concentrations of rat VLDL, most of the lipoprotein being removed by the heart. However, lipoproteins of density 1.019-1.063 g/ml were formed from human VLDL at all concentrations in the perfusate, as well as from higher concentrations of the rat VLDL. Agarose gel filtration of lipoproteins following heart perfusion with human VLDL revealed large aggregates containing particles which resemble low density lipoproteins (LDL) in electron microscopic appearance and apoprotein composition, since they contain largely apo-B. These data suggest that at normal concentrations rat VLDL are almost completely catabolised and taken up by the heart without the formation of LDL, while LDL is produced from human VLDL at all concentrations.


Catabolism of very low density lipoproteins in the rabbit. Effect of changing composition and pool size. To determine the metabolic mechanism of hypercholesterolemia in rabbits produced by feeding cholesterol-rich diets, control and hypercholesterolemic rabbits were injected with I-labelled very low density lipoproteins (VLDL, d 1.006 g/ml) from control and/or hypercholesterolemic donors. Apolipoprotein B in VLDL decayed biphasically. The first phase occurred much more rapid than the second. 95% of the VLDL apolipoprotein B was catabolized via the first phase (t1/2 = 0.55 +/- 0.19 h) in normal rabbit with the immediate appearance of this radioactivity in intermediate density lipoproteins (IDL, d 1.006-1.025 g/ml) and low density lipoproteins (LDL, d 1.025-1.063 g/ml). The apolipoproteins C and E at the same time were transferred to high density lipoproteins where they decayed biphasically. The apolipoprotein B from hypercholesterolemic VLDL in the normal recipient disappeared at a similar rate as from normal VLDL via phase I; however, it was incompletely converted to IDL and LDL. Apolipoprotein B from normal VLDL in cholesterol-fed rabbits disappeared at a normal rate via phase I, but only 82% was catabolized by this phase. Hypercholesterolemic VLDL injected into the hypercholesterolemic recipient was less rapidly catabolized via phase I (T1/2 = 2.5 +/- 0.89 H) and only a small fraction was converted to IDL and LDL.


Interactions of native and modified human low density lipoproteins with human skin fibroblasts. 125I-labeled low density lipoprotein (LDL) covalently bonded to Sepharose beads was not degraded by normal human fibroblasts nor did it trigger inhibition of sterol synthesis. The Sepharose beads loaded with LDL bound very tightly to the surface both of normal fibroblasts and fibroblasts from a subject with homozygous familial hypercholesterolemia; control Sepharose beads (activated sites covered with glycine) did not adhere to either cell type. LDL was extracted by a modification of the method of Gustafson (Gustafson, A. (1965) J. Lipid Res. 6, 512-517), so as to remove essentially all cholesterol, cholesterol ester and triglyceride. This modified LDL was bound, internalized and degraded as well as or better than native LDL. However, it failed to suppress sterol synthesis. These results provide additional evidence that the sterol moiety of the LDL is the key component affecting sterol synthesis. They also imply that the neutral lipids of LDL play a minor role in the binding of LDL to cell membranes and that the apoprotein rather than molecular size and shape is the critical factor.


Biochemical composition and heterogeneity of heparan sulfates isolated from AH-130 ascites hepatoma cells and fluid. The glycosaminoglycan composition of AH-130 ascites hepatoma cells and fluid were examined using enzymatic digestion, electrophoresis, and sequential partition fractionation. The cell-associated glycosaminoglycans were found to consist of 93% heparan sulfate, with the remainder consisting primarily of chondroitin sulfate. The glycosaminoglycans isolated from the ascitic fluid were found to consist of 58% heparan sulfate, 26% hyaluronic acid and 16% chondroitin sulfate. Dermatan sulfate was not detected in either cells or fluid. The heparan sulfate isolated from AH-130 cells in low-sulfate and highly heterogeneous with respect to biochemical composition. Fractions isolated by partition fractionation varied from 0.14 mol sulfate/mol uronic acid to 0.6 mol sulfate/mol uronic acid. Of the total sulfate 70--80% is N-sulfate in the former and 50% in the latter. Electrophoresis in 0.1 M HCl showed a highly heterogeneous material with mobility between that of hyaluronic acid and beef lung heparan sulfate. The heparan sulfate isolated from the fluid was similar to that isolated from the cells but was, however, somewhat more homogeneous with respect to charge.


Two types of cyclic GMP binding site associated with the cyclic AMP-dependent protein kinase from lymphocytes. Low- and high-affinity binding sites for cyclic GMP were found to be associated with the cyclic AMP-dependent protein kinase (ATP: protein phosphotransferase, EC 2.7.1.37) from human tonsillar lymphocytes, but neither of them was identical with the cyclic AMP binding site. The enzyme activated by cyclic GMP phosphorylated the same site of calf thymus H2b histone as the cyclic AMP activated enzyme; however, more complex kinetics of activation were found with cyclic GMP. Two classes of cyclic GMP binding site were demonstrated by kinetic analysis of cyclic [3H]GMP binding in the enzyme preparations eluted by 0.1 M potassium phosphate (pH 7.0) from DEAE cellulose. The high-affinity cyclic GMP binding site (Kd about 4 . 10(-8) M) belonged to some complex form of the protein kinase, as evidenced by the mutual inhibition of cyclic AMP binding and high affinity cyclic GMP binding. However, the high-affinity cyclic GMP binding site disappeared on Sephadex G-100 gel chromatography of the enzyme preparation, whereas the cyclic AMP binding activity was recovered quantitively as separate fractions. The low-affinity cyclic GMP binding site (Kd 2--5 . 10(-6) M) was demonstrated by the inhibitory effect of 10(-5) M cyclic GMP on cyclic AMP binding in each cyclic AMP binding fraction obtained by gel chromatography. However, cyclic AMP did not inhibit the binding of cyclic GMP to the low-affinity binding site.


Conformation of L-cystathionine, a carba analog of cystine, and stereochemistry of hormone-receptor interactions. The conformation of L-cystathionine, a carba analog of L-cystine, has been studied in the solid state using X-ray diffraction techniques. Crystal of L-cystathionine are tetragonal, space group P41 with cell constants a = 6.691(1) A, c = 21.998(3) A and Z = 4. From diffractometer data to the limit of 2theta = 162 degrees for Cukalpha, the structure was refined using full-matrix least-squares to an R value of 0.061. L-Cystathionine is isostructural chemically to L-cystine and its crystal structure is isomorphous to tetragonal L-cystine (Chaney, M.O. and Steinrauf, L.K. (1974) Acta Crystallogr. 1330, 711--716). The crystal structure of L-cystathionine is disordered, leading to two slightly differing conformers of L-cystathionine (each with half occupancy) with same helical sense but running in opposite directions and occupying the locations of L-cystine molecules in tetragonal L-cystine structure. Their conformational similarity, even when no sterical constraints such as cyclization are present, offers an explanation of the activities of the carba analogs of neuro-hypophysial hormones in terms of the structural integrity of the disulfide-like bridges.


Noninactivation of thyrotropin by cultured porcine thyroid cells. Freshly isolated porcine thyroid cells were cultured in the presence of highly purified porcine thyrotropin. Cells associate into follicles between the second and tenth day of culture and later form a monolayer. The biological and immunological activity of thyrotropin was measured daily in the media. Thyrotropin concentration and biological activity remained unchanged from the onset of the culture up to day 14. Limiting factors influencing thyroglobulin biosynthesis do not appear before day 13. The loss of follicular organization at day 10 cannot be explained by thyrotropin degradation in the medium. Considering the number of receptors per cell and the half life of the thyrotropin . receptor complex in the two dissociation compartments previously demonstrated, it appears in terms of both biological activity and affinity for the receptors that the thyrotropin molecules released from the first compartment do not differ from native molecules. It can be calculated that at least 31% of the molecules released from the second compartment are not inactivated. Thus, it is probable that the catabolism of thyrotropin on the receptor, or near the receptor site, does not play an important role in the regulation of thyroid cell function in vitro.


Mitogenic andadjuvant activities of polysaccharides from the cellular slime mold, Dictyostelium discoideum NC-4. Cell surface and extracellular polysaccharide fractions obtained from Dictyostelium discoideum NC-4 cultured in bacteria-free medium showed strong B-cell mitogenic activities. Upon periodate treatment of the extra-cellular polysaccharide fraction this activity completely disappeared. The extracellular polysaccharide fraction could also enhance the antibody response in vitro against sheep red blood cells.


Beta-adrenergic receptors and adenylate cyclase in hypertrophic and hyperplastic rat salivary glands. Isoproterenol induces both the secretion of protein and the stimulation of DNA synthesis and growth in rat salivary glands. The specific binding of the labelled beta-adrenergic antagonist [3H]dihydroalprenolol has been used to measure the number of beta-adrenergic receptors in rat parotid glands during isoproterenol-induced growth. Isoproterenol-enlarged glands display no change in the specific binding capacity per gland for [3H]-dihydroalprenolol compared with normal tissue. Catecholamine sensitive adenylate cyclase activity varies independently of the number of specific [3H]dihydroalprenolol binding sites during isoproterenol-induced growth. Previously-described di-ferences in optimal isoproterenol doses which produce protein secretion and stimulation of DNA synthesis may reflect different responses to various rates of receptor occupancy, or may be due to the presence of more than one type of beta-adrenergic receptor.


[Inhibition of the yeast respiratory system by Zn-protoporphyrin and effect of photolysis of this substance]. We have shown earlier that yeast cells grown in synthetic mediums supplemented with Zn++ accumulate large amounts of Zn-protoporphyrin within their mitochondria. This accumulation is accompanied by an inhibition of respiration (3). This study deals with the effect of light on the respiratory inhibition and the release of respiratory control which are observed if Zn-protoporphyrin is added to isolated mitochondria which are initially devoid of this pigment. In addition, we have studied the effect of light on the respiratory inhibition exerted by Zn-protoporphyrin accumulated in vivo. The following results were obtained: 1) The light-induced destruction of Zn-protoporphrin which had been added in vitro to Zn-protoporphyrin-free mitochondria significantly inhibits respiration and phosphorylation. Under these conditions, the extent of the inhibitions increases with the concentration of the added Zn-protoporphyrin and the duration of illumination. 2) Accumulation of Zn-protoporphyrin within the cells causes an inhibition of the respiratory activities and the activities of succinate-cytochrome c reductase and NADH-cytochrome c reductase of the mitochondria. Illumination of the isolated mitochondria from Zn-protoporphyrin-containing cells enhances the inhibition of these activities. No light-induced inhibition of these activities is observed with mitochondria from cells devoid of Zn-protoporphyrin.


[Origin and fixation of pyrophosphates in the parietal bones of young rats]. Calvaria from 6 to 13-day-old rats mineralize and bind pyrophosphates which are transferred from nucleotides, as shown by : 1 -- the increase of pyrophosphates in young rat calvaria incubated with nucleoside triphosphate, especially ATP ; 2 -- the more important increase of pyrophosphate content when ATP is renewed in the incubation medium ; and 3 -- binding of [32P] pyrophosphate [beta 32P] ATP. The effect of preheating of calvaria on 32P binding from [beta32P]ATP led us to assume that two systems might be involved in pyrophosphate fixation : a heat-labile, non-specific, phosphatase system, and a heat-stable pyrophosphotransferase system. cAMP increases the pyrophosphate content of calvaria incubated with ATP : that effect may result from either an inhibition of the phosphatase system, or an activation of the pyrophosphotransferase system.


[Registration of the Opaque-2 mutation by the EPR method]. The ESR method shows that variation of lysin concentration in the maiz enzyme resulting from the Opaque-2 mutation correlates with the number of paramagnetic centres in seed fractions. Increase of the lysin concentration brings about a quick rise of the ESR amplitude in the enzyme fraction of aleyrone layer and a decrease of the ESR amplitude in the seed germ.


[Model of a neuron-regulator of the effectiveness of synaptic transmission]. In diad and triad synapses of vertebrate retina the transmitter released from one and the same presynaptic membrane can act simultaneously on two (or even three) postsynaptic neurones. The model of such a diad synapse is proposed, in which the positive electrical feed-back is caried out between one of postsynaptic and presynaptic neurones. The feed-back appears because of potential drop across the longitudinal resistance of intercellular gap (RIII) near the activated synapse (see [1]). It is shown that if the input resistance of this postsynaptic neurone (N3) is low (low R3 in fig. 1,), it can operate as a regulator of effectiveness of synaptic transmission between the presynaptic (N1) and the second postsynaptic (N2) neurones. The effectiveness of synpatic transmission (e. g. the steepness of transfer function) is the more, the higher the membrane potential in the neurone acting as a regulator (fig. 2), and the higher the resistance of intercellular gap (RIII) (fig. 2). The model reproduces the effect of polarization of horizontal cells (by means of light or current) on the membrane potential in bipolars of the turtle and fish retina (fig. 3). The analysis of the model also shows that one of the functions of horizontal cells, as regulators of synaptic transmission between photoreceptors and bipolars, may be the detection of small objects and borders of image against uniform background.


[Mechanism of action of cyclic AMP-dependent histone kinase. Substrate specificity of the catalytic enzyme unit]. Interaction of several nucleotide derivates with homogenous catalytic subunit of cyclo-AMP-dependent histone kinase from pig brain is studied. Inhibition constants of these compounds are calculated, and the affinity of inhibitors to the enzyme active site is evaluated. The nature of heterocyclic base is found to be the main contribution into binding with substrate. The enzyme specificity with respect to a number of bivalent metal ions is studied, and Mg2+ is demonstrated to be the only efficient enzyme activator. It is shown by means of stationary kinetics that histone kinase-catalysed phosphotransferase reaction has a "ping-pong"-like mechanism.


[Effect of a single ethanol injection on lipid and lipoprotein synthesis in rat liver]. A single ethanol injection results in the increase of mono-, di- and tri-glicerides synthesis in rat liver, and also of the synthesis of apoprotein of very low density lipoproteins, their formation and secretion. Different uptake of pools of 14C-acetyl CoA, synthesized from injected 14C-acetate, and 3H-acetyl CoA, synthesized through metabolic pathways of 3H-leucine, indicates the compartmentalization of acetyl CoA in the synthesis of saturated and unsaturated fatty acids. 3H-acetyl CoA is more intensively used in the synthesis of unsaturated fatty acids than 14C-acetyl CoA synthesized from acetate. Ethanol increases the uptake of acetyl CoA, synthesized from acetate, for the synthesis of all the lipids, probably, for the expense of the increased synthesis of endogenous acetate in metabolic transformation of ethanol.


[Bioenergetics and proton-electron membrane systems]. Measurements of potential difference on energy producing subcellular particles by the methods of penetrating ions and of association of these particles with the planar membrane are described as well as some other studies of cell electrical energetics in which the author took part.


[Separation of catalytic and regulatory subunits of pigeon breast muscle protein kinase]. The separation of catalytic cAMP-independent and regulatory cAMP-binding subunits of pigeon breast muscle protein kinase was performed after the treatment of the holoenzyme by cAMP. The molecular weight of the homogeneous catalytic subunit is determined as 30,000. When catalytic and regulatory subunits were mixed, the decrease of phosphotransferase activity was observed.


[Conformation of cytochrome c, adsorbed on different substrates]. Visible spectra of cytochrome c adsorbed on solid supports and on the same supports covered with monolayers of lecithin and cholesterol were studied. It was shown that absorption spectra of high spin cytochrome c and pK of transition from low to high spin form depend on the nature of adsorbent surface.


REM sleep and the concept of vigilance.

REM sleep and the concept of vigilance. Several writers, notably Montague Ullman and Frederick Snyder, conceive of REM sleep and the associated state of vivid dreaming as periods of vigilance. In Ullman's conceptualization, the emphasis is on dreaming in humans, whereas Snyder is concerned with REM sleep (activated sleep) in subhuman organisms. In this paper several objections to the sentinel and vigilance theories are raised, and a modified concept of vigilance, linking it with learning and memory, is put forward.


Twenty-four-hour recording in REM-narcoleptics with special reference to nocturnal sleep disruption. Twenty narcoleptic patients and ten age-matched normals were polygraphically monitored for 58 consecutive hours. All subjects were on regimented sleep (hours between 2230 and 0700). Group A (11 patients and 10 normals) had enforced wakefulness during the day whereas Group B (9 patients) were permitted to sleep (mean = 2 1/2 hr.). On day 2, all subjects were permitted to sleep for 15-min periods every 2 hr. In narcoloptics, sleep recordings demonstrated a reduction of sleep latency, an increase of stage 1, and a decrease in stages 3 and 4 compared to normals, but total REM time and percentage of REM sleep were similar. Groups A and B showed no difference in the incidence of nocturnal awakenings. REM cyclic periodicity was larger in narcoleptics who also demonstrated a REM-sleep fragmentation. This fragmentation became more pronounced as time passed, with several shifts from REM to wakefulness and stage 1. Narcoleptics present REM onset sleep period but also show an inability to remain in REM sleep.


Paradoxical sleep and depth perception.

Paradoxical sleep and depth perception. It has been hypothesised that one possible function of paradoxical (REM) sleep is the maintenance of facilitation of co-ordinated eye movements. A prediction from this hypothesis is that binocular depth perception will be more accurate at the end of periods of paradoxical sleep than at the beginning. The results from previous studies are conflicting. Using two groups of eight healthy male volunteers in a two factor repeated measures design, it was found that for a period of paradoxical sleep in the second half of the night only, there was an improvement in binocular depth perception accuracy between the beginning and end of paradoxical sleep. The accuracy at the end of the paradoxical sleep was not significantly different to that on going to bed or on awakening in the morning; the effect was due to a large decrease in accuracy at the start of the REM period. There was no effect of paradoxical sleep on binocular depth perception in the early part of the night. Monocular depth perception accuracy was unaffected by paradoxical sleep.


Serum thymic factor and the neuromuscular transmission. The effect of Serum Thymic Factor (FTS) on neuromuscular transmission has been studied in vivo by electromyographic assay. As no modification of muscle tetanus pattern was detected, the involvement of this factor in the pathology of thymus disorders leading to myasthenia is unlikely.


Possible involvement of natural killer cells in bone marrow graft rejection. In the present study comparison between Natural killer (NK) cells and cells responsible for rejection of bone marrow grafts was made. Both cell populations were found to be inhibited by cyclophosphamide, silica, carrageenan, and C. parvum. The reactivity of both cell populations occurred late in life and was not expressed in infant mice. Mice tolerant to bone marrow grafts and, therefore, accepting parental marrow transplants showed also decreased NK cell reactivities. These common features between NK cells and bone marrow effector cells suggest but not prove that these cells could represent the same cell population.


[ATPase activity of the smooth and rough microsomal subfractions of the heart and liver in rabbits with a short-term microcirculatory disorder]. Short-time disturbances of the microcirculation in rabbits had a different influence on the ATP-ase activity of smooth and rough microsomes of the liver and heart. Mg+2ATP-ase activity in the liver decreased in both microsome fractions; as in the heart, the enzyme activity increased only in the smooth (light) microsomes.


[Adenosine deaminase activity in hepatomas of varying degrees of malignancy]. In highly malignant Gelshtein 22A hepatcma and ascites Ehrlich carcinoma adenosine deaminase activity was found to be reduced 3-fold as compared with that of the normal mouse liver. In less malignant hepatomas adenosine deaminase activity drops only by 20%. A certain reduction of adenosine deaminase activity was also noted in the liver to tumour-bearing mice.


[Cultivation of L strain mouse fibroblasts on silica gel plates]. A method for cultivation of mice fibroblasts, strain L, using silicagel slides, prepared with "KSK" silicagel, was developed Sterile, free of organic compounds, silicagel slides prepared for Petri dishes or test tubes can be successfully used as a solid medium promoting normal development of the culture.


Enhancement by oxotremorine of acetylcholine release from the rat phrenic nerve. Oxotremorine (10.5 micron) produced a paralytic effect on twitch responses of rat diaphragm in vitro to direct and indirect stimulation. 2 The paralytic effect of oxotremorine was absent when the diaphragm was stimulated directly in the presence of hemicholinium-3 (0.42 mM), at a time when twitch responses to indirect stimulation ceased completely. 3 Oxotremorine, at two different pharmacologically active doses, strikingly increased the resting as well as electrically evoked release of acetylcholine into the bathing fluid from the phrenic nerve-diaphragm preparation. 4 This presynaptic effect of oxotremorine may explain its pharmacological effects at the cholinergic synapses studied so far.


Responses of the pituitary-adrenal system of the pig to environmental changes and drugs. 1 The reactivity of the pituitary-adrenal axis of the young pig was tested for its suitability as a sensitive index for any discomfort that might be experienced under certain conditions of intensive husbandry.2 In a thermoneutral environment, most undisturbed piglets showed only slight variations in the plasma concentrations of adrenocorticotrophic hormone (ACTH) and corticosteroids.3 Stimuli such as exposure to ambient temperatures of +40 degrees C or -5 degrees C were required to cause large rises in the plasma concentrations of ACTH and corticosteroids.4 Apparently milder stimuli, such as change of environment, slight frustration or changes in ambient temperatures between +5 degrees C and +30 degrees C only rarely caused a significant rise in plasma corticosteroids. Thus changes in plasma corticosteroid concentrations are not a sensitive index for the reaction of a piglet to its environment.5 Increases in plasma ACTH concentrations occurred faster than those of the corticosteroids, were larger when expressed as a percentage of the basal values and occurred following relatively small disturbances such as omission of the reward in an operant behaviour test when corticosteroid changes were often not detectable. Thus rises in plasma ACTH might be a useful indication that a given situation is disturbing to a pig. The reaction of plasma ACTH concentrations to chronic irritations as they might occur in intensive husbandry remains to be investigated.6 Azaperone (2 mg/kg i.m.), a drug which is used as a sedative in pigs, caused a rise of about 50% in plasma corticosteroid concentrations. It did not diminish the large steroid output seen when the animals were exposed to high and low ambient temperatures.


Identification of functionally related neural assemblies. Present-day techniques of multiple-electrode together with computer-aided separation of impulses arising from  different neurons permit the simultaneous recording of nerve-impulse timings in sets of neurons exceeding 20 in number. This in turn makes it feasible to search for functional groups of neurons, defined as subsets that tend to fire in near simultaneity significantly more often than would independent neurons at corresponding mean rates. A statistical technique is described that permits  the detection and identification of such functional groups. The method is accretional, based on identification of associated neurons through interative application of a significance test on multiple coincidences of neuronal firings within an observational window. Examples of the operation of the method and indications as to its sensitivity are furnished through computer simulations of neural networks. The entire algorithm may be used as a screening technique to select smaller groups of neurons for cross-correlational and related finer-grained temporal analyses, or it may be used in its own right to detect and characterize functional groups that are not distinguishable by other statistical procedures.


Accumulation of cyclic adenosine 3',5'-monophosphate in human cerebellar cortex slices: effect of monoamine receptor agonists and antagonists. Noradrenaline, isoproterenol, adrenaline or clonidine stimulate cyclic AMP formation in human cerebellar cortex 15-20 fold and serotonin or dopamine 3-fold. The increase by noradrenaline or clonidine is dose-dependent. Propranolol completely blocks the increase induced by noradrenaline or isoproterenol but not that by clonidine. Phentolamine was ineffective in blocking cyclic AMP formation induced by noradrenaline, isoproterenol, adrenaline or clonidine. Chlorpromazine completely blocked the increase induced by noradrenaline, isoproterenol, clonidine, serotonin or dopamine.


[Nociceptive afferent blockade by percutaneous peripheral stimulation in the cat]. Experiments have been performed in order to study the effects of percutaneous peripheral stimulation (PCPS) both on the transmission of messages elicited by recruiting sensory units of the tooth pulp at the thalamic Centrum Medianum Level and on the jaw opening reflex (JOR). Both evoked potentials and JOR were inhibited by stimuli applied to the limbs by means of percutaneous (needle) electrodes. Observed inhibitory effects were not immediate: there was a latency period and progressive induction of these phenomena. The site of the inhibition is still unknown, nevertheless, the demonstration that PCPS was able to inhibit both evoked potentials in Centrum Medianum and JOR support the hypothesis that the analgesic effects may be due to descending inhibition blocking transmission of nociceptive information through the spinal cord.


[The effect of nalorphine on analgesia induced by peripheral stimulation]. The blockade of effects induced by percutaneous peripheral stimulation were abolished by injection of an opiate antagonist as nalorphine. Our results lead to the hypothesis that central and peripheral stimulations act by the same mechanism in producing blockade of noxious impulses. One may suggest that peripheral stimulations induce release of endogenous morphine-like substances which in turn give descending inhibition.


[Effect of differentiation of GABA levels in various cell cultures]. The effect of factors inducing somatic differentiation on cellular GABA level has been investigated. C6 glial cells from a rat astrocytoma present a significantly higher GABA level than the other glial or neuronal cells studied. A significant decrease in GABA levels in most cases parallels the somatic differentiation induced either by withdrawal of fetal serum, or by addition of dibutyryl cyclic AMP to the culture medium.


[Enhancement of compensatory renal hypertrophy by beta-1-24 corticotropin in the rat]. In the rat, the administration of beta1-24-corticotrophin during 7 days following an uninephrectomy enhances significantly the compensatory hypertrophy of the remaining kidney. There is no increase in renal compensatory hypertrophy when ACTH is injected to previously adrenalectomized rats. This action of ACTH could be related to the diabetes mellitus induced by this hormone or to an increase in sodium reabsorption by the tubular epithelial cells.


Bordetella pertussis in the National Capital Region: prevalent serotype and immunization status of patients. Over a 2-year period 67 strains of Bordetella pertussis were identified in 231 single specimens of nasopharyngeal secretions submitted from patients suspected to have whooping cough in the National Capital Region; 89.5% of the identifications were made by culture. Serotype 1,3 was predominant. At least 75% of the patients with bacteriologically confirmed whooping cough had not been fully immunized. There was no evidence that adenoviruses or other viruses played any important etiologic role in the 204 cases of whooping cough or whooping cough syndrome studied virologically.


Bronchiolar carcinoma: report of 11 cases and review of the literature. Eleven cases of bronchiolar carcinoma seen between 1970 and 1975 are reviewed. Roentgenographic manifestations at the time of initial examination included single peripheral nodules, multiple nodules, irregular cavities and a persistent infiltrative form. Microscopic tissue examinations showed two patterns of tumor growth: a papillary form resembling tall columnar cells extending along but not invading the alveolar septal walls and a cribriform pattern represented by inflammatory thickening of the alveolar septal walls and pleomorphic cellular proliferation within luminal spaces. The papillary form was associated with the cavitary lesions. Long term survival is dependent on early recognition of the single peripheral nodule, cavity or small persistent infiltrative form managed by a thoracotomy and lobectomy.


Acute leukemia complicating treatment of glioblastoma multiforme. A five-year-old girl developed acute myelomonocytic leukemia after fifteen months of intensive chemotherapy and irradiation for glioblastoma multiforme. The leukemia became manifest while the patient was in a remarkable remission brought about by treatment with high-dose methotrexate with citrovorum rescue. This is the first reported association of these disorders in the same patient. It is possible that the leukemia was induced by the treatment, since both radiation and the chemotherapeutic drugs used have been shown to be leukemogenic in some circumstances. The patient developed leukemia in a setting of relatively normal peripheral blood counts, having had very little myelosuppression from her treatment.


Oat cell carcinoma (apudoma) of the esophagus: a case report. An oat cell carcinoma occurring in the esophagus of a 62-year-old man is described. A strong argyrophilia of the oat cells together with the presence electron microscopically of neurosecretory type granules in their cytoplasm suggests a derivation of this tumor from the cells of the APUD series, and the tumor is diagnosed as apudoma. An aberrant columnar epithelium on the esophageal surface is presumed to be the source of its origin. A rapid postoperative course of the patient despite a rather circumscribed growth pattern of the primary tumor appears to characterize the clinical aspect of this case. An apparent accumulation of the cases with esophageal apudoma in Japan is discussed.


Cyclic nucleotide concentrations in 1,2-dimethylhydrazine induced rat colon adenocarcinoma. The intracellular concentrations of adenosine 3',5'-cyclic monophosphate (cAMP) in colonic tumors induced in adult male Holtzman rats by 1,2-dimethylhydrazine (DMH) were found to be 1/2 the concentration found in normal large bowel tissue. Intracellular concentrations of guanosine 3'-5'-cyclic monophosphate (cGMP) in the neoplastic cells were twice the normal colon level. The concentrations of these two cyclic nucleotides were relatively constant throughout the normal colon. Thus, the anomalous tumor cyclic nucleotide concentrations are attributed to the specific cell population of the lesion and not to the site of development within the colon.


Globin messenger RNA content in hepatomas: a test of retrogenesis. The globin mRNA content of foetal, neonatal and normal adult liver and of the transplantable Morris hepatoma 5123C and host liver of animals carrying the tumour was measured using molecular hybridization of total nucleic acid extracts from these tissues with a complementary DNA copy of globin mRNA from rabbit reticulocytes. The purpose of these experiments was to determine, as a test at the molecular level of the retrogenesis hypothesis, if the gene for foetal globin is activated in neoplastic hepatic tissue or induced in host liver. These experiments did not detect any foetal globin mRNA in the hepatoma or host liver nucleic acid extracts in spite of the increase in haemopoietic activity induced in host animals as a consequence of tumour bearing.


Equivalent expression of endogenous murine leukemia virus-related genes in C3H/10T1/2 cells and chemically transformed derivative cells. The possibility that chemical carcinogens may induce enhanced expression of endogenous C-type RNA tumor virus genes in the absence of intact virus particle production has been partially tested in a model system. Thie was accomplished by measuring the abundance and diversity of murine leukemia virus-related RNA sequences associated with the polyribosome fraction of nontransformed C3H/10T1/2 clone 8 cells and a 3-methylcholanthrene-transformed derivative clone. Although both clones are virus nonproducers, they were found to contain significant amounts of polyadenylate-containing murine leukemia virus-related RNA sequences; however, both the types and quantities of such sequences appear indistinguishable in both clones. These results suggest that expression of the corresponding gene sequences into RNA is not related to the maintenance of the transformed state in these chemically transformed cells.


A comparison of methods for the isolation of carcinoembryonic antigen. Carcinoembryonic antigen (CEA) was isolated from a human tumor with 0.02 M sodium phosphate containing 0.14 M NaCl (pH 7.0) (saline) and further studied after treatment with perchloric acid or 8 M urea. Preparations CEA obtained from saline homogenates and both methods of treatment were characterized by isoelectric focusing and gel filtration. Perchloric acid treatment resulted in an approximate 10-fold decrease in protein and a significant loss of CEA as compared to the saline- and urea-treated material. Isoelectric focusing revealed that the resultant CEA subpopulations were dependent upon the method of isolation. Urea- and saline-treated material showed complex isoelectric patterns that were quantitatively dissimilar. Perchloric acid-treated material showed a comparatively simple isoelectric pattern that was not significantly affected by electrofocusing in the presence of urea. Gel filtration on ACA 34 of the CEA obtained from each method of isolation resulted in two peaks of activity. The first peak corresponded to the void volume of the column, and the second peak coeluted with commercially available purified 125I-labeled CEA. Centrifugation of the peaks obtained resulted in a significantly greater loss of CEA from the void peak of each isolation procedure. The amount of CEA lost from the void peaks following centrifugation differed with each method of isolation and suggested the presence of aggregates.


Differential inhibition of the rejoining of X-ray-induced DNA strand breaks in normal and transformed human fibroblasts treated with 1,3-bis(2-chloroethyl)-1-nitrosourea in vitro. The effects of 1,3-bis(2-chloroethyl)-1-nitrosourea on the rejoining of X-ray-induced DNA strand breaks were examined in normal human fibroblasts (WI-38) and a simian virus 40-transformed derivative (VA-13) with the use of alkaline sucrose sedimentation. 1,3-Bis(2-chloroethyl)-1-nitrosourea was capable of partially inhibiting repair of X-ray-produced DNA strand breaks in both cell types when the drug was added to the culture medium immediately after X-irradiation. However, when 1,3-bis(2-chloroethyl)-1-nitrosourea exposure preceded X-ray by 1 hr, DNA repair was inhibited to a much greater extent than it was when 1,3-bis(2-chloroethyl)-1-nitrosourea followed X-ray. The inhibition of DNA repair by 1,3-bis(2-chloroethyl)-1-nitrosourea appeared to be complete in the transformed VA-13 cells, while only partial inhibition of repair was observed in the normal WI-38 cells.


Activation of C-type virus during chemically induced leukemogenesis in mice. Repeated percutaneous applications of 7,12-dimethylbenz(a)anthracene on weaning DBA/2 and ST/a mice induced 100% leukemias with short latency periods. Endogenous C-type viruses were activated during the treatment as evidenced by (a) increased expression of the murine leukemia virus major core protein, p30, in blood and spleens and (b) increased frequency of detection of ecotropic virus by cocultivation of the splenocytes with SC-1 cells. The treatment did not affect p30 expression in several nonlymphoid organs, and detection of xenotropic viruses in the splenocytes was decreased. Virus expression did not correlate with the progression of disease in that (a) high p30 levels were generally found in mice with relatively low spleen weights and (b) p30 levels had no obvious connection to survival of the individual. 7,12-Dimethylbenz(a)anthracene treatment had little influence on p30 expression in spleens and blood from C3H and BALB/c mice, which are less sensitive to 7,12-dimethylbenz(a)anthracene-induced leukemogenesis. The results indicate an association of C-type virus activation with chemical induction of leukemia but do not necessarily imply an etiological role of the virus in the disease.


Cyclic adenosine 3':5'-monophosphate and methotrexate transport in L1210 cells. Treatment of L1210 cells with glucose, 1-methyl-3-isobutylxanthine, theophylline, and prostaglandins E1 and E2 was used to increase to varying degrees the endogenous level of cyclic adenosine 3':5'-monophosphate (cAMP). In an inverse manner these agents decreased the rate of methotrexate (MTX) transport into the cells. Azide, conversely, lowered the cAMP level and enhanced MTX transport. Quantitative information about this relationship was obtained by plotting the reciprocal rates of MTX transport as a function of cAMP levels. These data suggest that MTX transport is regulated by the level of cAMP.


Oxidative phosphorylation and aspects of calcium metabolism in myocardia of hypercholesterolaemic swine with moderate coronary atherosclerosis. Aspects of myocardial oxidative phosphorylation and Ca2+ metabolism were studied in a swine model in which coronary atherosclerosis was induced by a combination of denudation of the endothelium of the coronary arteries plus 7--11 months of feeding a high fat--high cholesterol diet. By microscopy, a moderate amount of coronary atherosclerosis was present at the time of sacrifice, and 2 of the 14 swine hearts had old myocardial infarcts. Myocardial mitochondria from grossly normal areas showed partial uncoupling and decreased state 3 O2 uptake with 3 of 4 substrates tested. In addition, Ca2+ stimulated mitochondrial respiration was decreased in the atherosclerotic swine. In the sarcoplasmic reticulum Ca2+ uptake under conditions of heavy loading was greater in the atherosclerotic swine than in control animals. The degree of atherosclerosis was not great enough to suggest that persistent myocardial ischaemia was present. Possibly coronary artery spasm induced an intermittent ischaemia resulting in the metabolic abnormalities observed, or the changes may have been brought about by the effects of the high fat--high cholesterol diet on subcellular membranes.