Electron spin polarization in photosynthesis and the mechanism of electron transfer in photosystem I. Experimental observations. Transient electron paramagnetic resonance (EPR) methods are used to examine the spin populations of the light-induced radicals produced in spinach chloroplasts, photosystem I particles, and Chlorella pyrenoidosa. We observe both emission and enhanced absorption within the hyperfine structure of the EPR spectrum of P700+, the photooxidized reaction-center chlorophyll radical (Signal I). By using flow gradients or magnetic fields to orient the chloroplasts in the Zeeman field, we are able to influence both the magnitude and sign of the spin polarization. Identification of the polarized radical and P700+ is consistent with the effects of inhibitors, excitation light intensity and wavelength, redox potential, and fractionation of the membranes. The EPR signal of the polarized P700+ radical displays a 30% narrower line width than P700+ after spin relaxation. This suggests a magnetic interaction between P700+ and its reduced (paramagnetic) acceptor, which leads to a collapse of the P700+ hyperfine structure. Narrowing of the spectrum is evident only in the spectrum of polarized P700+, because prompt electron transfer rapidly separates the radical pair. Evidence of cross-relaxation between the adjacent radicals suggests the existence of an exchange interaction. The results indicate that polarization is produced by a radical pair mechanism between P700+ and the reduced primary acceptor of photosystem I. The orientation dependence of the spin polarization of P700+ is due to the g-tensor anisotropy of the acceptor radical to which it is exchange-coupled. The EPR spectrum of P700+ is virtually isotropic once the adjacent acceptor radical has passed the photoionized electron to a later, more remote acceptor molecule. This interpretation implies that the acceptor radical has g-tensor anisotropy significantly greater than the width of the hyperfine field on P700+ and that the acceptor is oriented with its smallest g-tensor axis along the normal to the thylakoid membranes. Both the ferredoxin-like iron-sulfur centers and the X- species observed directly by EPR at low temperatures have g-tensor anisotropy large enough to produce the observed spin polarization; however, studies on oriented chloroplasts show that the bound ferredoxin centers do not have this orientation of their g tensors. In contrast, X- is aligned with its smallest g-tensor axis predominantly normal to the plane of the thylakoid membranes. This is the same orientation predicted for the acceptor radical based on analysis of the spin polarization of P700+, and indicates that the species responsible for the anisotropy of the polarized P700+ spectrum is probably X-. The dark EPR Signal II is shown to possess anisotropic hyperfine structure (and possibly g-tensor anisotropy), which serves as a good indicator of the extent of membrane alignment.


[Concentration of free fatty acids in muscle following administration of corticotropin and hydrocortisone]. Experiments were conducted on rats; the gas chromatographic method was applied to the study of the free fatty acids content in the gastrocnemius 30 minutes after the intraperitoneal injection of adrenocorticotropic hormone (ACTH)--1 Unit per 100 g and hydrocortisone acetate--1 mg per 100 gm of body weight. It was shown that in the resting muscles ACTH increased the content of stearic acid, whereas hydrocortisone--of both stearic oleic acids. The changes in the content of other free fatty acids were insignificant. During the short-term activity the content of stearic acid in the regimen of single rhythmic contractions in the gastrocnemius of intact rats increased. In experiments with ACTH and hydrocortisone this elevation was much less and not significant. ACTH and hydrocortisone stimulated the stearic acid consumption by the muscles during the activity.


[Post-radiation disorders in the cyclic AMP system of mouse spleen and thymus lymphoid cells]. Early alterations in the enzyme activities controlling cyclic adenosine-3',5'-monophosphate (cAMP) metabolism after the irradiation (800 rad) of mice were found in the lymphoid cells of the spleen and thymus. Postradiation disturbances in these activities' ration induced alterations in cAMP steady-state concentrations in the cell. It was also demonstrated that irradiation reduced lymphoid cell ability to accumulate cAMP in response to isoproterenol administration.


[Effect of colchamine on Ehrlich ascitic carcinoma cells showing synchronized cell division]. The author studied the 24-hour changes in the number of normal and colchamine mitoses in the cells of Ehrlich's ascites carcinoma in mice after the injection of colchamine argainst the background of partial synchronization of cell division, obtained as a result of preliminary injection of dibutyryl cyclic 3',5'-AMP, and also in mice after the injection of colchamine alone or dibutyryl cyclic 3',5'-AMP. As shown, synchronization of cell division in the tumour led to the 2,6-fold increase in the number of tumour cells blocked by colchamine and also to the accelerated arrest of colchamine mitoses.


[Effect of thyroliberin (thyrotropin-releasing-hormone) on secretion of thyrotropic hormone and prolacting in a monolayer cultrue of rat adenohypophysis]. Cells of the adenohypophysis in the primary 5--8 day monolayer culture responded to the administration of the thyrotropin-releasing-hormone (TRH) by a rapid dose-dependent release of thyrotropic hormone (TTH) and prolactin into the culture medium. This effect is independent of the serum content in the nutrient medium. Thyroxin, the thyroid gland hormone, blocks the stimulating action of TRH with respect to the TTH secretion, but not to prolactin. The blocking effect of thyroxin is apparently expressed not on the cell membranes, but in the cytoplasm, distal to cAMP, by way of the hormonal signal transmission.


[Comparative study of several structuro-functional parameters of the liver and adrenal cortex in mice of different genetic strains under physiologically normal conditions]. In adult male mice CBA and C57BL there were revealed differences in the morphometric characteristics of the subcellular organization of hepatocytes, as well as in the indices of functional capacity of the adrenal cortex and the rate of metabolism of steroid hormones in the liver. In animals of the strains under study the levels of steroid hormones production by the adrenal cortex and the rate of gross metabolism of steroid hormones in the liver were characterized by inverse relationship. Structural-functional indices of the adrenal cortex and the liver are supposed to cause different liver responses in these strains of mice to the influence of the alterating factors.


Response of blood vessels to sympathetic nerve stimulation. The frequency response curves (FRC) of isolated blood vessels differ from each other not only in their initial slopes, but in their maxima, and their intercept on the frequency axis. Within the physiological range, there is a linear relation between response and frequency. The FRC of most vessels with junctional innervation are similar, varying only somewhat with innervation density. Other factors found to influence the FRC are: variation in innervation distribution, in the extent of myogenic propragation, and large differences in the sensitivity of the alpha-adrenergic receptor. The effectiveness of the transmitter increases with frequency rise. The maximum effective radius of the transmitter released from one varicosity is only several microns. In vessels with light to moderate innervation density at low frequencies, there is probably little overlap of transmitter effect from adjacent varicosities even at the outermost layers of smooth muscle cells. There is a disparity between neurogenic response and that which might be expected from the direct action of the transmitter. It is proposed that a local limited myogenic extension of excitation may extend the local action of a quantum of transmitter.


Modes of vasoconstrictor and vasodilator neurotransmission. Vasoconstrictor and vasodilator neuroeffector transmission occurs in a variety of modes. Models are presented depicting vascular segments under the direct control of a single or multiple innervation, or an indirect influence via secondary release of a vasoactive substance. The neuroeffector relationship in the rabbit portal vein is described to illustrate the coexistence of several modes of transmission. ATP or its congener is released from the nonadrenergic vasodilator nerves in this vein, possibly as the transmitter. Further, a similar substance is released from the adrenergic vasoconstrictor nerves. It possibly acts directly on smooth muscle cells to relax them, and on the nerve terminals to inhibit the adrenergic transmitter release. Evidence suggests that such purine-mediated feedback mechanism may also occur at many other adrenergic and nonadrenergic neuroeffector synapses.


Relationship between noradrenaline-induced depolarization and contraction in vascular smooth muscle. In strips of rabbit main pulmonary artery, full dose-response curves of noradrenaline for the mechanical and electrical responses of the vascular smooth muscle cells were obtained under normal conditions and after increasing and decreasing the membrane potential with strychnine and tetraethylammonium, respectively. The results suggest that the magnitude of the noradrenaline-induced contraction is related to the height of the membrane potential and not to the degree of depolarization produced by the adrenergic transmitter.


Involvement of calcium in cyclic nucleotide metabolism in human vascular smooth muscle. When cyclic nucleotide phosphodiesterase was purified from isolated smooth muscle layer of human aorta by DEAE-cellulose column chromatography, separated cyclic GMP phosphodiesterase activity was markedly stimulated in the presence of 10-20 micrometer of Ca2+ by a protein modulator which has similar physicochemical properties to troponin C. Synthetic compound, N-(6-aminohexl)-5-chloro-1-naphthalensulfonamide, which produced relaxations of arteries contracted by prostaglandin F2alpha or KCl was found to inhibit selectively this Ca2+-dependent cyclic GMP phosphodiesterase. This compound produced inhibition of superprecipitation of myosin B system obtained from mouse skeletal muscle and also inhibited adenosine triphosphatase activity of myosin B. Our data suggest that calcium is involved through a protein modulator in cyclic nucleotide metabolism of vascular smooth muscle and that the calcium-dependent protein modulator probably participates in the regulation of contractile response of vascular smooth muscle by affecting actomyosin ATPase activity.


[Hormonal factors in the prediction of hormonedependency of breast cancer]. Biological diagnostic of hormonedependency of breast tumors is important for the choice of treatment. Modifications of the metabolism of steroids, oestrogens, androgens, and glucocorticoids allowed a first approach. More recently, investigations have been made on breast tissue receptors for hormones that are involved in the growth of the normal gland. It is actually suggested that one could predict the hormonedependency of a tumor when it contains both receptors for oestradiol and progesterone.


Serum concentrations of 25-hydroxy vitamin D in Europeans and Asians after oral vitamin D3. Serum concentrations of 25-hydroxy vitamin D (25-OHD3) were measured in seven Asians of Indian extraction and eight Europeans before and at intervals after taking 1 mg vitamin D3 by mouth. In all subjects the concentrations rose in the 24 hours after ingestion. There was little change over the next nine days in the concentrations in the Europeans but those in the Asians continued to rise until about day 10. Subsequent rates of fall in 25-OHD3 were similar in the two groups. Our observations suggest that the low serum concentrations of 25-OHD3 found in Asians are not caused by either impaired intestinal absorption of vitamin D or rapid clearance of 25-ODH3 from the plasma.


Outbreak of Guillain-Barré syndrome associated with water pollution. Sixteen cases of Guillain-Barré syndrome occurred in the third week of a diarrhoea epidemic caused by water pollution in EL-Sult, Jordan. Of 30 000 people exposed to polluted water, 5000 developed diarrhoea, 74 typhoid, and 30 infectious hepatitis. Thirteen of the 16 patients with Guillain-Barré syndrome had been mildly affected by diarrhoea 8-24 days before the onset of peripheral neuropathy. Paralysis progressed rapidly, reaching a peak in one to five days, and recovery began three to seven days after the start of the most severe symptoms. All but four patients had recovered completely after one year. Rapid progress of paralysis and a delayed interval between maximum weakness and start of recovery were both associated with poor prognosis.


Sterol ordering effects and permeability regulation in phosphatidylcholine bilayers. A comparison of ESR spin-probe data from oriented multilamellae and dispersions. The effects of a series of sterols on molecular order and motion in bilayers formed from egg lecithin and dicetylphosphate were examined and correlations between order and data on permeability to 22Na+ were sought. Electron spin resonance spectra were observed for probes intercalated both in multilamellar dispersions where the effects of motion and orientation are difficult to separate, and in planar multibilayers where the degree of molecular order may be measured even in the presence of slow probe motion. It was concluded from the planar multi-bilayer data that sterols which increase the degree of ordering of lipid molecules decrease 22Na+ permeability, and that sterols which have the opposite effect on order increase permeability. All the sterols tested lead to decreased rates of motion of the probes. This effect obscures the correlation between order and permeability using data from dispersions.


The purification and characterization of a DNA nicking-closing enzyme from Bacillus megaterium. Although several eucaryote DNA nicking--closing (N--C) enzymes have been characterized, only the Escherichia coli enzyme has been extensively studied amongst procaryotes. The latter enzyme is distinctly different from the eucaryotic enzymes and we have therefore purified the N--C enzyme from Bacillus megaterium to determine if procaryotes form a distinctive class of N--C enzymes. The purified B. megaterium N--C enzyme has a molecular weight of 120,000, only partly relaxes negative supercoils, does not affect positive supercoils, requires Mg2+, and is inhibited by 0.2 M KCl. The enzyme is also inhibited by 1 mM nalidixic or oxolinic acids but unaffected by novobiocin. A crude N--C enzyme preparation from Micrococcus luteus shows very similar properties.


DNA nicking--closing activity from salmon testis. Salmon testis is a good source of DNA nicking--closing (N--C) enzyme, as expected for rapidly proliferating cells. Partial purification was obtained but it resulted in an unstable form of N--C enzyme. Cruder fractions are useful for relaxing supercoiled DNA under conditions where other N--C enzymes are inactive. The enzyme is very tolerant of salt concentration, with activity still detectable at 0.6 M NaCl. It is also active at low temperatures with reasonable activity at 0 degrees C. However, at temperatures greater than 25 degrees C it is rapidly inactivated. Otherwise, its properties are similar to other eucaryotic N--C enzymes.


Density and protein profiles of myelin from two regions of young and adult rat CNS. Myelin, isolated from forebrain and spinal cord of young and adult rats, was distributed by zonal centrifugation on linear (0.32--1.00 M) sucrose gradients in a bell-shaped mode. The peak position of forebrain myelin shifted from the density of 0.58 M sucrose in young animals to that of 0.67 M sucrose in adult rats, while in spinal cord no such pronounced shift was noticed (approximately 0.58 M sucrose). Morphologically, the preparations appeared very similar across the density ranges. Specific activities of acetylcholinesterase were substantially below the total homogenates, while those of 2', 3'-cyclic nucleotide 3'-phosphohydrolase were higher in all fractions, except in the light myelin subfractions from adult spinal cord. Basic proteins decreased from the light to the heavier fractions; higher molecular weight proteins increased, together with proteolipid protein, which in spinal cord reached a plateau and in forebrain decreased towards the heavy side. The ratio of the small basic protein/large basic protein showed higher values in the light myelin subfractions in the regions and ages examined, pointing to a higher degree of maturation.


A new look at pituitary adenomas: structure elucidating function. Cases of seven different types of surgically resected pituitary adenoma are described. Included are tumours secreting prolactin or growth hormone or both, and nonfunctioning tumours--undifferentiated and oncocytic tumours, and one tumour with cells of the adrenocorticotropin-melanocyte-stimulating hormone type. The final interpretation of a case of pituitary adenoma should include an assessment of thorough morphologic studies, using not only routine staining and light microscopy but also immunostaining and electron microscopy, to complement the biochemical, radiologic and clinical evaluation.


Effect of a chronic tryptophan dietary deficiency on the rat's sleep-wake cycle. The sleep-wake cycle of 12 tryptophan dietary deficient rats and their non-deficient paired controls were observed for a 8:00 a.m. to 8:00 p.m. period. EEG, EMG and body activity were continuously monitored on polygraphic recordings throughout the 12 hr observation period. The results indicate no significant difference between the tryptophan deficient and sufficient animals in time spent awake, slow-wave or parodoxical sleep. There was a non-significant trend among the tryptophan deficient animals to be less active and spend more time in both slow-wave and paradoxical sleep, which is in contrast to an expected insomnia effect. The results do not support the suggested relationship between reduced serotonin levels and the occurrence of insomnia, questioning the serotonergic theory of sleep.


Contribution of angiography to the diagnosis, staging and assessment of radiation and chemotherapy of solid abdominal malignancies in children. Sequential angiographic studies were done in six children to stage and assess the results of radiation and/or chemotherapy of solid abdominal malignancies: one bilateral Wilms' tumor, two neuroblastomas, two hepatoblastomas and one hepatocarcinoma. Angiography was of value in demonstrating the tumor, its location, extent and vascular characteristics, as well as its regression and recurrence. Wilms' tumor and neuroblastoma responded and well to radiation and chemotherapy with substantial decrease in tumor size and regression or disappearance of tumor neovasculature. Resceted tumors revealed this to be due to tumor necrosis, hemorrhage and/or cystic degeneration. Hepatoblastoma and hepatocarcinoma did not respond as well to chemotherapy, with only mild decrease in size and neovasculature of the tumor.


Radiation therapy in the treatment of malignant salivary gland tumors. A retrospective analysis of 52 patients with malignant salivary gland tumors is reported. Seventeen patients received early postoperative radiation therapy and 16 (94%) were free of local or regional disease 2-14 years following initiation of therapy, although 14 were considered at high risk of developing local recurrence. Two subjects (12%) developed distant metastases and 14 (82%) were completely disease-free. Survival and disease-free status of patients treated for recurrent or inoperable disease were much worse with two of 13 disease-free at 45 and 168 months respectively. Various workers have reported recurrence rates after surgery along at 25-38% and over 50% for many histological types. On the basis of this report early postoperative radiation therapy is recommended to reduce the risk of postsurgical recurrence. Prognostic trends relating to both histological type and location of primary disease are discussed.


Malignant transformation of esophageal columnar epithelium. This report describes two patients with esophageal columnar epithelium (Barrett's esophagus) in which microinvasive adenocarcinoma developed. Case 1 had multiple foci of carcinoma in situ contiguous with epithelium resembling gastric and intestinal mucosa. Case 2 had signet-ring type adenocarcinoma. Surveillance for malignant transformation in columnar esophageal epithelium should be routinely performed, and because of its focal nature, multiple biopsies and cytologic examination should be carried out. The presence of carcinoma in situ should lead to consideration of excision of the affected esophagus.


Malignant fibrous histiocytoma and pleomorphic sarcoma in association with medullary bone infarcts. A malignant fibrous histiocytoma arose in the proimal tibia of a 40-year-old man who had multiple, symmetrically distributed, medullary bone infarcts of unknown etiology, involving the distal femora and the tibiae. Despite amputation and chemotherapy, widespread metastases developed and death occurred 19 months after surgery. A polemorphic sarcoma, probably representing an anaplastic malignant fibrous histiocytoma, arose in association with a single medullary infarct in the proximal humerus of a 33-year-old woman. She remains well without evidence of disease five years after treatment by radical radiotherapy followed by shoulder disarticulation. Sarcoma arising in association with bone infarction is a rare entity. Sixteen cases reported in the medical literature, including our own, are reviewed. The sarcomas arose in the tibia in nine cases, the femur in six cases and the humerus in one case. The patients were usually older individuals and 13 of the 16 were male. All but two had multiple bone infarctions. Four of the patients had caisson disease, three had what is probably an hereditary bone dysplasia, one had sickle cell disease and eight had infarcts of unknown etiology. Most patients have had a rapidly fatal outcome. Thirteen of the sarcomas have been fibrosarcomas or malignant fibrous histiocytomas, both of which are rare primary bone tumors. Analysis of the published cases of bone infarction-related sarcomas suggests that the risk of developing a sarcoma is greatest in infarcts with large medullary components.


Malignant clear cell acrospiroma.

Malignant clear cell acrospiroma. The histologic features of a malignant clear cell acrospiroma were those of infiltrative local growth, frequent mitoses and angiolymphatic invasion. The histochemical and ultrastructural findings were similar to those reported for benign clear cell acrospriomas. Amputation of the leg and regional node dissection were required for clinical control. Evaluation of this case and review of the literature suggests that the malignant clear cell acrospiroma often behaves in an aggressive manner and frequently metastasizes. As a consequence, thereapeutic strategies should be appropriately planned.


Immunohistochemical demonstration of alpha-1-antitrypsin in the islet cells of human pancreas. Alpha-1-antitrypsin was demonstrated both by immunofluorescence and immunoperoxidase techniques in the peripheral islet cells of 15 normal human pancreases, processed freshly and after embedding in paraffin. Normal human liver obtained from the same patients was negative. The immunohistochemical reactions were stronger in frozen sections than in paraffin material. However, immunoperoxidase staining on paraffin sections permitted a more precise localization of positive cells. The specificity of the immunohistochemical reactions was confirmed by applying various control tests including a absorption of the specific antisera with purified alpha-1-antitrypsin, inhibition and blocking tests. Further examination of pancreatic tissue for the presence of various immunoglobulins, alpha-1-lipoproteins, alpha-1-acid glycoproteins and ceruloplasmin were negative. These findings suggest that the pancreatic islets may be an extra source of alpha-1-antitrypsin.


Cytological observations on the blood and hemopoietic tissue in the crab, Callinectes sapidus. I. The fine structure of hemocytes from intermolt animals. A fine structural analysis of hemocytes recovered from the pericardial sinus of the blue crab (Callinectes sapidus) demonstrates that hyaline cells, intermediate cells, and granulocytes are present in the hemolymph. A detailed consideration of their shared and differing cytological features suggests that the various blood cells represent different stages of cytogenesis along a common or single path of cellular differentiation. The potential functional complexity of the hemocytes is attested to by the occurrence within their cytoplasm of three and/or possibly four morphologically distinct types of inclusion bodies, and other membrane-bounded compartments (e.g., lysosome-like structures, autophagic vacuoles, peroxisome-like bodies) whose function is unknown. Preliminary examination of the well-developed Golgi organelle in the intermediate cells and granulocytes indicates its probable participation in the elaboration of the inclusion bodies; however, the details of its role in granulogenesis and in the known plurifunctional capacities of these cells remain obscure.


Glycogenolysis in amphibian liver organ cultures: effects of isoprenaline, cyclic-AMP, calcium, and the ionophores A23187 and Br-X537A. Isoprenaline, db-cAMP and ionophore Br-X537A induced glycogenolysis and glucose release from A. means liver organ cultures in the presence or absence of medium Ca2+. Ionophore A23187 had similar effects, but only in the presence of external Ca2+.


Changes in the content of cyclic AMP and cyclic GMP in eggs of Mactra solidissima during the second cleavage cycle. The levels of cyclic AMP and cyclic GMP during the second cleavage cycle of eggs of Mactra solidissima have been measured. Cyclic GMP levels vary reciprocally with cyclic AMP levels, the cyclic GMP level increasing up to 4 to 5-fold and the cyclic AMP level falling to 50 percent at the onset of mitosis.


Phenotypic modification of SV40-transformed hamster lymphoid cells in vivo. Explants of simian virus 40 (SV40)-induced lymphoid tumors yield SV40-T-antigen-positive derivatives that differ from GD248 lymphocytes propagated in suspension culture, (or in vivo), in the following respects: polygonal shape, adhesion to culture substrates in vitro, phagocytic capacity, lack of immunoglobulin and a chromosome complement at least twice that of GD248 lymphocytes. When GD-248 lymphocytes are propagated as suspension in vitro, no such adherent variants can be detected. However, sequential in vivo passage of GD248 lymphocytes obtained from the suspension-culture lines also yield adherent cell lines upon explanation in vitro. Injection of adherent cells into hamsters produces tumors with histological features of reticulum cell sarcoma.


Inhibition of internuclear transport of SV40-induced T antigen in heterokaryons. Internuclear migration of tumour specific nuclear T antigen has been analysed in SV40-induced H-50 tumour and chick erythrocyte heterokaryons. Thirty hours after cell fusion the incorporated and enlarged erythrocyte nuclei were invariably T antigen-positive. When treated with colchicine at 10(-4) or 10(-7) M concentration, the erythrocyte nuclei incorporated into the heterokaryons did not swell and remained T antigen-negative. The results strongly suggest the involvement of a colchicine sensitive contractile protein matrix in the internuclear transport of T antigen and other proteins.


Cyclic GMP regulation and responses of Polysphondylium violaceum to chemoattractants. In cells of the cellular slime mold Polysphondylium violaceum an attractant, which is released during the aggregation stage, causes a transient rise of the cyclic GMP concentration. Cells of this organism develop in shaken suspensions after they have finished growth. Cell development is not accompanied by an increase in the EDTA stability of cell adhesion. Both the developmental regulation and the specificity of chemotactic responses is reflected in the light scattering patterns recorded in cell suspensions: Folic acid causes a strong response in early preaggregation cells and the Polysphondylium attractant does the same in aggregation competent cells, whereas cyclic AMP is inactive in both stages.


Cyclic-AMP stimulated calcium influx into aggregating cells of Dictyostelium discoideum. Within about 10 seconds after stimulation of Dictyostelium discoideum cells with cyclic AMP an increased rate of 45Ca influx was observed. Part of the cellular calcium reappeared in the extra-cellular medium between 1 and 3 minutes after stimulation. No effect of 5'AMP on calcium distribution was found. The transient calcium influx is discussed in connection with chemotaxis and other cyclic-AMP induced responses.


Endomyocardial disease and eosinophilia. Report of a case. While an association between blood eosinophilia and endomyocardial disease has been recognized, the role of the eosinophil in the pathogenesis of the cardiac lesions remains uncertain. In a 69-year-old-man with large cell carcinoma of the lung, marked eosinophilia was stimulated by and progressed with the course of the neoplasm which was producing an eosinophil chemotactic factor. Peripheral blood eosinophils were vacuolated and degranulated while those in the bone marrow were morphologically normal. Clinical evidence of cardiac dysfunction developed one month prior to death. At autopsy, 12 months after the onset of symptoms, endomyocardial disease was present. There were numerous eosinophils in the damaged myocardium and surrounding the pulmonary neoplasm. In patients with endomyocardial disease and eosinophilia, the eosinophil may be directly cardiotoxic or a primary mediator of cardiac damage; therapeutic attempts to reduce the number of eosinophils might be benefit.


Enzyme immunoassay for human apolipoprotein B, the major protein moiety in low-density- and very-low-density lipoproteins. We used enzyme immunoassay to measure apolipoprotein B concentration in human plasma. Pure lipoprotein B was isolated from serum samples of fasting normolipidemic subjects by sequential preparative ultracentrifugation and coated to a polystyrene tube surface by adsorption. Human serum samples and rabbit antiserum to human apolipoprotein B were incubated with the solid-phase lipoprotein B. Soluble antigen competed with solid-phase antigen for binding to antibodies. After washing, peroxidase-labeled sheep antibodies against rabbit immunoglobulins were added, and after further washing the bound label was assayed. This provided a direct measurement of the soluble antigen. The best technical conditions for the assay were determined. The minimum detectable concentration was 1 microgram per assay. The enzyme immunoassay yielded values that compare favorably with those obtained by radial immunodiffusion (r = 0.84) and by rocket immunoelectrophoresis (r = 0.80). The assay offers several advantages over existing techniques: sensitivity, specificity, simplicity, ane non-use of radioisotopes.


Adenylate kinase inhibition by adenosine 5'-monophosphate and fluoride in the determination of creatine kinase activity. The current methods for the determination of creatine kinase (EC 2.7.3.2) activity are derived from Oliver's method, in which AMP is used to decrease interference by adenylate kinase (EC 2.7.4.3). Recently, Szasz et al. and Rosano et al. described methods in which diadenosine pentaphosphate and fluoride, respectively, are used to reduce this interference. However, diadenosine pentaphosphate does not sufficiently inhibit such activity of hepatic origin, while fluoride alone can only inhibit it at concentrations at which the fluoride tends to precipitate as MgF2. Finally, Szasz et al., the Committee on Enzymes of the Scandinavian Society for Clinical Chemistry and Clinical Physiology, and the German Society for Clinical Chemistry have proposed methods in which both AMP and diadenosine pentaphosphate are used to inhibit adenylate kinase. We have found that by using low concentrations of AMP and fluoride together, we can greatly diminish this interference without significant loss of creatine kinase activity and with no precipitation of MgF2.


Nephropathia epidemica: immunohistochemical study of pathogenesis. Renal immunohistochemistry was studied in 21 patients with Nephropathia epidemica, a benign hemorrhagic fever endemic in northern Europe. Between days 4 and 25 after the onset of fever, scanty but distinct deposits containing immunoglobulins and C3 were detected in glomeruli and tubules. The occurrence of deposits was verified with electron microscopy. It is concluded that an immune complex mechanism plays a part in the pathogenesis of this disease. It is also evident that the febrile period coincides with the presence of immune complexes in the circulation.


Gastric calcification detected in vivo by 99mTc-pyrophosphate imaging. A 48-year-old white man with Hodgkin's disease was referred for bone imaging. There was intense accumulation of the bone imaging agent in the stomach and in the region of the upper lobe of the right lung. Subsequent biopsy and histological examination revealed that there was amorphous calcifications in the mucosal layer of the stomach. Possible causes for this calcification are discussed.


Uptake by bone of pyrophosphate, diphosphonates and their technetium derivatives. 1. The uptake of inorganic pyrophosphate (PPi) from blood to bone was investigated in the rat in vivo. 2. PPi is taken up by the bone, where it appears both as PPi and as inorganic orthophosphate (Pi). The latter is due at least partly to local hydrolysis. 3. The fraction of injected PPi taken up by bone, measured as total PPi, was in the same range as that of technetium-tin-PPi, diphosphonates, technetium-tin-ethane-1-hydroxy-1,1-diphosphonate and Pi, but lower than that of calcium. 4. The plasma half-life of PPi is in the same order of magnitude as that of technetium-tin-PPi, diphosphonates, technetium-tin-ethane-1-hydroxy-1,1-diphosphonate, Pi and calcium. 5. PPi, diphosphonates and their technetium complexes are only partly ultrafiltrable in plasma. 6. It appears that the technetium complexes behave in a similar fashion to free PPi or diphosphonate.


The effect of alpha-adrenoreceptor stimulation on the airways of normal and asthmatic man. 1. The bronchial response to alpha-adrenoreceptor stimulation has been investigated in normal and asthmatic subjects with a specific alpha-receptor agonist, methoxamine hydrochloride, after atropine and beta-adrenergic blockade. 2. No significant changes in forced expiratory volume in 1 s (FEV1.0) were seen in normal subjects. 3. Methoxamine inhalation caused mild symptomatic wheezing and significant falls in FEV1.0 in asthmatic subjects. No change in FEV1.0 occurred after inhalation of distilled water as control. 4. It therefore appears that methoxamine causes bronchoconstriction in asthmatic subjects through its alpha-adrenergic-stimulating properties, since we were not able to detect any beta-receptor-blocking activity of methoxamine on the airways.


Impact of aftercare services on recidivism of mental hospital patients. The subjects in this study were 129 patients released within a 2-year period from a large state hospital to a three-county area. Patient information on demographic, inhospital, and posthospital variables and the extent of participation in an aftercare program was collected from existing records. The relationship between the community support system available to the patient and recidivism was determined. Three criteria of recidivism were used: readmission within 1 year following discharge, number of days in the community within 1 year following discharge, and number of days to first readmission. No single predictor was adequate in predicting hospital readmission. When the effects of confounding variables were eliminated using multiple-regression techniques, the amount of aftercare received was an important factor in predicting recidivism. However, current situational factors appeared to be more important predictors of recidivism than receipt of aftercare services.


Development of an aftercare program in a nonmetropolitan area. This article describes the development of a nontraditional aftercare program set in a predominantly rural area. The program was initiated by a citizen's board concerned with the lack of follow-up services provided to patients discharged from a distant state hospital. The planning format as conceived by representatives of this board is outlined and the steps involved in operationalizing specific services and a research program are described. Lastly, a number of limitations of the program and future directions are enumerated.


Effects of Yunnan Bai Yao on blood platelets: an ultrastructural study. The effects of Yunnan Bai Yao on blood platelets were studied at an ultrastructural level. This herbal drug is found to cause the release of platelet constituents which accounts for its hemostatic effects. The fine structural changes of platelets in response to this drug are also described.


Diagnosis of equine neurologic problems. This is a review of the more common diseases of the spinal cord and various areas of the brain of horses. The results of a two and one-half year study of spinal cord disease are emphasized. After a description of the lesion the salient clinical signs are described and the features that differentiate them from other similar diseases. In the seminar, films of case and slides of lesions will be shown to document these diseases.


Clinicopathological manifestations of selected neoplasms. This report provides a general overview of the pathobiology of neoplasia, and an update on the clinicopathological manifestations of lymphosarcoma, mastocytoma, histiocytoma, melanoma, sarcoid and circumanal gland tumors in domestic animals. Neoplasia represents a continuum of events from reversible hyperplasia to irreversible and pathological changes in tissue growth patterns. In some instances the causes of this disease process have been identified, but the etiology of the majority of naturally occurring neoplasms remain unknown. Surgical excision is the preferred treatment for tumors, but is often more beneficial when combined with chemotherapy, radiotherapy or immunotherapy. The successful diagnosis and management of neoplastic disease in domestic animals necessitates a thorough awareness of the clinical presentation and biologic behavior of specific tumors on the part of veterinarians who provide health care for these species.


The diagnostic laboratory and its relationship to practitioners. Diagnostic laboratory professional staff. Diagnostic services provided to practitioners include necropsy, histopathology, bacteriology, virology, parasitology, immunopathology, clinical pathology and toxicology. Services are also available at the college in avian and aquatic diseases. The laboratory expects to continually upgrade its services through the addition of appropriate tests, re-evaluation of existing tests, development of methods for more convenient specimen collection and shipment, increased responsiveness to veterinarians' needs, epidemiological investigations, and continuing education.


The canine contagious respiratory disease complex (kennel cough). Several infectious agents are involved in the kennel cough complex in dogs. They include canine parainfluenza virus (SV5), canine adenovirus 2, Bordetella bronchiseptica, and possibly several mycoplasma species. The importance of each of these agents in the disease syndrome is discussed as well as possible prevention or treatment.


Pituitary--adrenal responsiveness of rat mothers to noxious stimuli and stimuli produced by pups. Studies on the response of female rats to various stresses during the course of lactation showed a marked reduction in pituitary-adrenal activity. Maximum suppression coincided with the period of maximum lactation, about 14 days after parturition. Both pituitary corticotropin and the release of corticosterone were significantly reduced. Females showing maternal behaviour but not lactating did not exhibit this buffering of the stress response. Another aspect of pituitary-adrenal activity has been studied in lactating female rats. Females, although they have a buffered stress response, responded differently, in terms of pituitary-adrenal activity, to stimuli emitted by the pups: pups which are exposed to noxious stimuli elicit a much greater pituitary-adrenal response in lactating females than do pups which are merely handled. Separation from the pups does not elicit a pituitary-adrenal response. This difference in response is modified if the infants are malnourished and is not observed in virgin animals that are not lactating.


Purine biosynthesis in mutant mammalian cells. De novo purine biosynthesis has been studied in lymphocyte cell lines established from Lesch-Nyhan patients deficient in hypoxanthine-guanine phosphoribosyltransferase (HGPRT), in in vitro differentiating erythroleukaemic cell lines cloned from cells charactistic of virus-induced murine leukaemia, and in mutant hamster cells deficient in amidophosphoribosyltransferase. The relationship between cellular phosphoribosylpyrophosphate (PP-ribose-P) metabolism and the activity of the enzymes which catalyse the early steps of de novo purine biosynthesis has been explored. It was found that hamster cells deficient in amidophosphoribosyltransferase did not accumulate PP-ribose-P as do HGPRT-deficient cells. In these model systems, an accelerated rate of de novo purine biosynthesis tended to be associated with an increase in cellular PP-ribose-P cotent, but decreases in this rate results from the reduction in the activity of amidophosphoribosyltransferase. Regulation of ammonia-dependent de novo purine biosynthesis was similar to that of glutamine-dependent purine biosynthesis.


Superactivity of phosphoribosylpyrophosphate synthetase, due to feedback resistance, causing purine overproduction and gout. A mutant feedback-resistant, physiologically superactive, phosphoribosylpyrophosphate (PP-ribose-P) synthetase was found in a family with purine overproduction, gout and uric acid lithiasis. In haemolysates and cultured fibroblasts from the propositus, the mutant enzyme exhibited resistance to feedback inhibition by normal cell constituents, such as ADP and GDP; normal affinity to substrates and to activator Pi was demonstrated in the haemolysate. In both erythrocytes and cultured fibroblasts, the superactivity of the mutant enzyme was manifest in increased PP-ribose-P content and availability for nucleotide synthesis, leading to an acceleration of the rate of purine synthesis de novo in the fibroblasts. The enzyme abnormality and the resulting increase in PP-ribose-P content and generation were demonstrated in the erythrocytes of one of the propositus' two siblings who was similarly affected but not in the propositus' father, his second brother and four sons, who were all clinically and biochemically normal, nor in the erythrocytes of the clinically normal hyperuricosuric mother. However, cultured fibroblasts from her skin exhibited variability in PP-ribose-P content and availability and in the rate of purine synthesis de novo, these parameters being increased in most cultures. The mother's fibroblast cultures were found to contain two cell populations, one with normal and the other with mutant PP-ribose-P synthetase, indicating an X-linked pattern of inheritance of the synthetase superactivity in this gouty family.


Purine metabolism and control of cell proliferation. Exposure of normal lymphocytes to phytohaemagglutinin or other lectin mitogens results in increased concentrations of 5-phosphoribosyl-1-pyrophosphate (PP-ribose-P) within minutes. Subsequently, synthesis of purine nucleotides by both the de novo and the salvage pathways is facilitated. This change is prevented by proliferation-inhibiting concentrations of exogenous adenosine. The capacity of lymphocytes to metabolize both adenine and adenosine is increased several-fold by incubation with phytohaemagglutinin but the specific activities of the respective first-step enzymes are not significantly altered. These results suggest that the relatively low quantity of PP-ribose-P available in normal lymphocytes is a major factor limiting the synthesis of purine nucleotides and may be important for the maintenance of the quiescent state. Increased availability of PP-ribose-P may also be associated with proliferative activation of fibroblast-like cells: chick embryo fibroblast cultures released from density-dependent inhibition of growth by insulin, trypsin or serum rapidly increase the rate of adenine incorporation into nucleotides. Chick embryo fibroblasts transformed by Rous sarcoma virus, but not cells infected with the respective non-transforming leukosis virus, show PP-ribose-P concentrations higher than those observed in normal cells.


Microbial models and regulatory elements in the control of purine metabolism. Bacterial systems have been used to identify and characterize the organization of the genetic units and the regulatory elements that control purine metabolism. An analysis of 13 genes that control the biosynthesis of AMP and GMP has revealed three multigenic operons. These show properties of gene contiguity, promoter sites, coordinate expression and polarity effects. The unit controlling the formation of IMP is one operon (pur JHD) consisting of three genes which together control the formation of phosphoribosylglycinamide synthetase (EC 6.3.4.13), an early enzyme in the biosynthetic pathway, and a terminal bifunctional complex (IMP cyclohydrolase--formyltransferase). Regulatory mutants were isolated and characterized by several methods including the use of a unique fusion of two unrelated operons. Both operator constitutive and repressor type (purR) mutations have been identified. The purR product functions in the common control of several genetically distinct enzymes that participate before the formation of IMP. Plasmid DNA enriched for the purE operon has been isolated and used in the study of the role of nucleotide effectors in the binding of repressor-like proteins. AMP but not GMP is needed for binding, and purR mutants are deficient in the binding substance. Mutants with differential blocks in the salvage and interconverting reactions have been used to characterize the regulatory elements of the formation and the activity of guanosine kinase, GMP reductase (EC 1.6.6.8), and purine nucleoside phosphorylase (EC 2.4.2.1). Two structural gene products (purF) and (purG) have been implicated as possible regulatory elements for the use of guanosine, and a role for glutamine in the induction of GMP reductase has been revealed.


The effect of adenosine on lymphoid cell proliferation and antibody formation. The discovery of an association of certain primary defects in human purine metabolism with immunodeficiency disease has served to focus attention on the possible role of purine compounds in the functional activity of lymphoic cells. Considerable evidence has accumulated of the need within the intact organism for purine and pyrimidine compounds supplied by the liver as a supplementary nutritional requirement for the growth of the rapidly proliferating tissues of certain organ systems. Likewise, lymphoid cells cultured in vitro show evidence of an enhancement of indices of cellular proliferation and of antibody synthesis when exogenous adenosine is added to the medium. These functions are inhibited by high concentrations of adenosine and there is some evidence that T-cell proliferation shows a greater sensitivity to inhibition by adenosine than B-cells. These observations may be significant in relation to the known defects in human purine metabolism and their mechanism for producing immunodeficiency.


Purine and pyrimidine metabolism: pathways, pitfalls and perturbations. The conceptual framework which underlies many studies of purine and pyrimidine metabolism in intact cells has been critically evaluated. The model that is implicit in many such studies is the single, partially purified enzyme. This paper gives examples both of instances in which the extrapolation of results of enzymes studies to intact cells has been successful and of instances in which enzymes behave differently in the intact cell than in cell extracts. Pitfalls in the extrapolation of results of enzyme studies to intact cells concern (a) metabolic pathways, (b) intracellular enzyme activities, (c) enzyme regulation, and (d) intracellular metabolite concentrations. Examples are also given of situations in which perturbations in one aspect of purine or pyrimidine metabolism lead to changes in other aspects, often distant in the network of reactions.


Effect of bilateral superior cervical ganglionectomy (SCG) on pituitary adrenal function in the male albino rat. The effect of sympathetic denervation (bilateral superior cervical ganglionectomy) on several aspects of pituitary adrenal function was studied in 250-300g rats. Group I=Sham-op, Group II=superior cervical ganglionectomy. (1) There was no significant difference between these groups in the plasma corticosterone (B) response to ether or immobilization stress or to ACTH administration. (2) Circadian periodicity of plasma B was determined by sequential (q.4h) tail vein sampling under LD (lights on 0800) off 2000) and under DL (lights on 2000, off 0800). Both groups showed similar peak and trough concentrations. Phase shifting occurred in both groups at day 11. (3) Compensatory adrenal hypertrophy of similar magnitude was present in both groups 10 days following unilateral adrenalectomy. These studies indicate that sympathetic input from the superior cervical ganglia is not involved in stress induced, circadian or some feedback aspects of pituitary adrenal function.


Prolactin receptors in mammary tumors of GR mice. Prolactin receptors have been identified in estrone-progesterone induced mammary tumors from GR mice. 125I-labeled ovine prolactin binding to tumor homogenates reached a steady state in 12 hours at 22 degrees and was specific for prolactin. Prolactin receptors were highest (16 fmoles/mg protein) in primary, hormone-dependent tumors and declined progressively in transplanted hormone-dependent and transplanted hormone-responsive tumors. In autonomous tumors, binding was approximately 5% of that found in primary tumors. Scatchard analysis of binding to selected tumors indicated that the observed decrease in bound hormone was due to a loss in the number of receptor sites; binding affinity was unaltered (kd approximately 1 X 10(-10) M). Since receptors for estrogen and progesterone as well as those for prolactin decline in a concerted manner with the transition to autonomy, autonomous growth may result from a loss of receptors or an increase in the relative proportion of autonomous cells present in the tumor.


Postnatal development of gonadotropin binding sites and cAMP synthesis in ovaries and estradiol plasma levels in estrogenized or androgenized female rats. The binding of 125I-HCG by ovarian homogenates was studied in adolescent rats injected with 100 microgram estradiol dipropionate or 50 microgram testosterone propionate at the age of three days. Estrogenized and androgenized female rats showed a decrease of gonadotropin binding in ovaries at the 25th postnatal day. After the day 40, however, there was an increase in gonadotropin binding capacity above that in control animals. The changes of specific binding of 125I-HCG by ovaries of rats neonatally treated with sex hormones did not correlate with either the responsiveness of cAMP synthesis in ovaries to LH or with plasma levels of estradiol-17beta in postnatal development.


The long search for the causes of congenital malformations in mammals. The history of teratology is reviewed. Methods available for causing malformations in mammals by subtle changes in the environment during pregnancy are discussed. A table is provided in which the commoner teratogens are listed. Methods are described by means of which the activity of some common teratogens may be opposed. The occurrence at Battle Creek, Michigan, USA, of a chemical agent dangerous to health, life and development of all mammals is described. This gives an indication that injury and death to all mammals, as in the Sevaso disaster, including teratogenesis may be expected to be a fairly frequent occurrence in the years to come.


Translation of ascites and mengovirus RNA in fractionated cell-free systems from uninfected and mengovirus-infected Ehrlich-ascites-tumor cells. We have prepared homologous, fractionated, cell-free translational systems from uninfected and mengovirus-infected Ehrlich ascites tumor cells in order to determine what alterations occur following virus infection in the translational machinery of the host cell. Two major differences distinguish the system developed from infected cells. First, it has a 40% lower rate of protein synthesis, primarily a consequence of the rate of chain elongation, which is depressed to 60 amino acids/min from 90 amino acids/min in the system from uninfected cells. Second, at supraoptimal concentrations of Mg2+ and K+ the system from virus-infected cells supports the translation of mengovirus RNA but not host mRNA. These differences between the two systems may reflect specific changes which are responsible for the selective translation of mengovirus RNA in the infected cell. In both systems the optimal concentrations of polyamines, monovalent and divalent cations, mRNA, and ribosomal subunits are the same for the translation of either host or viral RNA. This uniformity is useful in experiments, designed to investigate the selective translation of viral RNA, where various components of the two systems are interchanged.


Selective translation of mengovirus RNA over Host mRNA in homologous, fractionated, cell-free translational systems from Ehrlich-ascites-tumor cells. The selective translation of viral RNA in mengovirus-infected Ehrlich ascites tumor cells was investigated using fractionated translational systems whose macromolecular components were derived entirely from uninfected or virus-infected cells. Both systems translate host mRNA from uninfected cells, host mRNA from virus-infected cells, and mengovirus RNA. In competition experiments, where viral RNA and host mRNA were translated together in systems from uninfected cells, the relative amounts of virus-specific and host-specific proteins synthesized were proportional to the relative concentrations of the RNA templates. In systems whose components were obtained from virus-infected cells, mengovirus RNA was preferentially translated. 70% of the selectivity found in the translational systems derived from infected cells was due to the initiation factor fraction, the remaining 30% to components of the pH 5 enzyme fraction. In addition, host mRNA isolated after virus infection is translated in vitro to a lower extent in the presence of mengovirus RNA than is host mRNA from uninfected cells.


Mechanism of activation of protein kinase I from rabbit skeletal muscle. Mapping of the cAMP site by spin-labeled cyclic nucleotides. Binding of adenosine 3':5'-monophosphate (cAMP) to protein kinase (type I) from rabbit skeletal muscle has been investigated using spin-labeled cAMP derivatives. Different compounds were synthesized with the spin label attached by spacer chains of different length at different positions on the adenine base. Immobilization of the spin label, determined by comparing the electron-spin resonance spectra recorded in the presence of the kinase with those of the free ligand in solutions of different viscosities, gave information about the geometry of the cAMP site. Strong immobilization of the N-6 substituents up to a spacer length of seven atoms indicates a rather deep cleft of the cAMP site. The depth of this cleft differs, however, when the spin label is attached to the different positions at the adenine (N-6, C-2 and C-8). Whereas the N-6 derivatives indicate a rather deep site, the C-2 derivatives reveal a significantly smaller depth and C-8 substituents (syn conformation) obviously occupy a very shallow surface with almost no immobilation. In addition the binding affinities of the spin-labeled cAMP derivatives have been determined, together with those of a series of (diamagnetic) C-2 derivatives bearing hydrophobic alkyl chains of different length. The latter results helped to clarify the differences between the regions near to C-2 and N-6, respectively, of the cAMP site. N-6 spin-labeled derivatives have also been investigated in the presence of ATP and protein kinase. These results are interpreted as indicative of a conformational change at the cAMP site upon formation of the holoenzyme, due to binding of ATP, leaving cAMP less strongly immobilized.


Pyrophosphate-stimulated uptake of calcium into the germlings of Phytophthora infestans. The uptake of 1 micrometer calcium into 6-h-old germination tubes of the fungus Phytophthora infestans follows Michaelis-Menten kinetics with a Km of 33 +/- 4 micrometer and a V of 0.3 nmol.min-1.(5 x 10(4) cells)-1. Uptake is inhibited by ruthenium red and lanthanum (both at 1 micrometer) and by the proton conductors 2,4-dinitrophenol (1 mM) and carbonylcyanide m-chlorophenylhydrazone and carbonylcyanide p-trifluoromethoxyphenylhydrazone (1--10 micrometer) and also by sodium azide. These data suggest that calcium uptake is dependent on energy and on a carrier. Calcium uptake is stimulated by pyrophosphate but not by ATP, orthophosphate, or polyphosphate. This stimulation is prevented by proton conductors or by incubation at 0 degrees C.


Circular dichroic properties and conformation of thionicotinamide dinucleotides. The spectroscopic properties of the 3-thioamide analogues of coenzymes NAD and NADH (sNAD and sNADH) have been investigated in order to obtain information about their conformational properties. In particular, ultraviolet absorption and circular dichroism properties of solutions in phosphate buffer pH 7 and ethanol were studied. Also equimolar mixtures of AMP and sNMN(H), obtained by cleaving the coenzymes with phosphodiesterase, were investigated using the same solvents. The appearance of a couplet around 260 nm, which is not present for the mixture of sNMN and AMP, suggests a stacking interaction of the two aromatic moieties in sNAD. This conclusion is further substantiated by a hyperchroism of the ultraviolet absorption band in the 260-nm region in both sNAD and sNADH. The comparison of the ultraviolet and circular dichroic properties of intact and cleaved coenzymes in the different solvent systems makes it possible to single out the bands which are more sensitive to conformation changes (i.e. to open-stacking equilibrium) and those which are sensitive to solvent effects only.


Circular dichroic properties and conformation of thionicotinamide dinucleotides bound to horse-liver alcohol dehydrogenase. The interaction between horse liver alcohol dehydrogenase and the oxidized and reduced forms of the 3-thionicotinamide--adenine dinucleotide coenzyme analogues (sNAD and sNADH) has been investigated by ultraviolet absorption, fluorescence and circular dichroism. The fluorescence of sNADH is enhanced when bound to the enzyme, and the protein fluorescence is quenched by both sNADH (60--65%) and sNAD (65%). The possible origin of the larger quenching produced by sNAD with respect to that of NAD is discussed. Coenzyme dissociation constants have been determined by monitoring the quenching of the protein fluorescence, and some kinetic consequences of these dissociation constants are discussed. The dichroic properties of various enzyme complexes have been investigated, and are discussed in terms of conformational changes and environmental changes during coenzyme binding. The conformation of sNAD bound to the enzyme in the presence of trifluorethanol and the conformation of sNADH bound to the enzyme in the presence of isobutyramide have been analyzed in particular detail. Also the circular dichroic spectrum of the apoenzyme is discussed in terms of contributions of the aromatic amino acid residues in the highly resolved 240--310-nm region and in terms of helix content in the 220-nm region.


Fluorescence properties of reduced thionicotinamide--adenine dinucleotide and of its complex with octopine dehydrogenase. Reduced 3-thionicotinamide--adenine dinucleotide (sNADH) is shown to be fluorescent, with an emission maximum at 510 nm when excited in the region of the absorption maximum (398 nm), and with a very low quantum yield, (3.4 +/- 0.5) x 10(-4). The interaction between sNADH and octopine dehydrogenase was investigated by ultraviolet-difference spectroscopy and fluorescence. Some surprising fluorescence features were found when sNADH was bound to the enzyme in the presence of D-octopine, as follows. (a) There is an unusually high enhancement of the dinucleotide fluorescence (by at least a factor of 100) attended by a 40-nm blue shift of the emission maximum. (b) The protein fluorescence is quenched almost completely. (c) The bound coenzyme analog undergoes a photoreaction, which proceeds differently from that occurring the free form. These features appear to be unique to the octopine.sNADH complex, as for example they are not present when sNADH is bound to horse liver alcohol dehydrogenase, or when NADH is bound to octopine dehydrogenase. The possible origin of these fluorescence features is discussed. Binding and kinetic studies were carried out with sNAD and sNADH. It was found that sNAD neither binds nor acts kinetically as a coenzyme. sNADH exhibits relatively good binding, with Km and Ki values close to those of the natural coenzyme, but the turnover number is 460 times smaller than that with NADH. The kinetic consequences of these findings are discussed. The sNADH dissociation constants were determined as a function of temperature, and appear to be practically temperature-independent in the range 10--40 degrees C. It seems thus, in agreement with previous studies, that the interaction between octopine dehydrogenase and coenzymes proceeds athermically, regardless of the structure, affinity, and chemical reactivity of the coenzyme. The possible biological and chemical meaning of this finding is discussed.


Fatty acid beta-oxidation system in microbodies of n-alkane-grown Candida tropicalis. Localization of fatty acid beta-oxidation system in microbodies of Candida tropicalis cells growing on n-alkanes was studied. Microbodies isolated from the yeast cells showed palmitate-dependent activities of NAD reduction, acetyl-CoA formation and oxygen consumption. When sodium azide, an inhibitor of catalase, was added to the system, palmitate-dependent formation of hydrogen peroxide was observed. Stoichiometric study revealed that two moles of NAD were reduced per one mole of oxygen consumed in the absence of sodium azide and the presence of the inhibitor doubled the oxygen consumption by microbodies without an appreciable change in NAD reduction. These results indicate that the yeast microbodies contain beta-oxidation system of fatty acid, and that catalase located in the organelles participates in the degradation of hydrogen peroxide to be formed at the step of dehydrogenation of acyl-CoA.


Comparison of viable and nonviable tumor uptake of Sc-46, Mn-54, Zn-65, In-111 and Au-195 with Ga-67 citrate in a hepatoma model. The tissue distribution of Sc-46, Mn-54, Zn-65, In-111 and Au-195 were studied in a rat hepatoma model at various time intervals over a 96 h period. The tumor localizing properties of these isotopes were evaluated by examining their incorporation and clearance from viable and nonviable tumor tissue and determining the critical tissue ratios formed with blood and muscle. In general, the results showed greater uptake in viable than nonviable tumor tissue at early time periods (4-24 h). By 96 h, however, the activity remaining in the nonviable tumor tissue exceeded the quantity in viable tumor tissue. This trend was previously noted for Ga-67. When compared with Ga-67, only Mn-54 among the isotopes studied showed remarkably higher viable tumor/blood ratios (4-24 h, 45:1-83:1 respectively). Manganese-54 also showed highly significant accumulation in cardiac muscle with a heart/blood ratio at 4 h superior to comparable values previously reported for Cs-137 and Tl-201. It is suggested that tumor and heart imaging may be feasible utilizing radioactive manganese (Mn-51 or Mn-52) with the new positron imaging systems.


Triple radioisotope technique in etiologic evaluation of space-occupying lesions of the liver. The findings from 43 patients with histologically proven focal liver disease examined by triple isotope tracer study are presented. The method consisted in a three step technique, the first step being an isotope scan after injection of 99mTc-sulfur colloid and 131I rose bengal. As a second step dynamic hepatic scintiangiography was carried out with 99mTc human serum albumin in order to assess the vascularity of a lesion detected on the initial scan. Thirdly, the patient was administered 67Ga to evaluate further the pathologic process. According to our results this triple radioisotope technique provides the clinician with valuable information on the possible etiology of focal liver disease.


Cyclic AMP in cereborspinal fluid of rats: effects of electroconvulsive shock. The concentration of cyclic adenosine 3',5'-monophosphate (cAMP) was measured in cisternal cerebrospinal fluid (CSF) of male and female rats after electroconvulsive shock. cAMP levels were found to be increased compared to sham-treated controls for several hours following electroconvulsive shock with maximal increases of 46.8% in male and 17.2% in female rats. Old rats (300--360 g body weight) showed higher baseline concentrations than younger rats (180--250 g). In both groups female rats tended to have higher cAMP levels than male rats.


Cholestyramine and ileal by-pass in the treatment of familial hypercholesterolaemia. A comparison was made of the therapeutic effectiveness of cholestyramine and an ileal by-pass operation as hypocholesterolaemic measures in thirteen patients with familial xanthomatotic type II hypercholesterolaemia. Serum cholesterol and faecal steroids were measured before and at the end of a 10 day course of cholestyramine (32 g/day), and subsequently after an ileal by-pass operation. The mean decrease in serum cholesterol caused by cholestyramine (-17%) was significantly less than that caused by the surgical procedure (-33%). The increase in faecal steroid excretion, mainly as bile acids, was lower with cholestyramine (1.261 g/day) than after the ileal exclusion (2.176 g/day) and a positive correlation was found between the decrease in serum cholesterol and the increase in the faecal elimination of steroids of cholesterol origin. However, even though the increase in faecal steroids with cholestyramine was positively correlated with that resulting from ileal by-pass, the correlation between the corresponding changes in serum cholesterol level was not significant. The findings indicate that ileal by-pass decreases serum cholesterol and increases faecal elimination of cholesterol more effectively than cholestyramine, and that the decrease in the serum cholesterol level induced by ileal exclusion is not consistently predictable by the serum cholesterol response to the preceding cholestyramine treatment.


Serum immunoreactive prolyl hydroxylase in liver disease. The concentration of serum immunoreactive prolyl hydroxylase (SIRPH) was measured in thirty patients with chronic active hepatitis, thirteen with primary biliary cirrhosis, four with alcoholic or idiopathic cirrhosis, and four with acute hepatitis; the values were compared with those in twenty-three control subjects. Increases in SIRPH were found in all the groups with liver diseases, individual values being highest in primary biliary cirrhosis in which about two-thirds of patients had values more than two standard deviations above the mean value in the control subjects. No correlation was found between SIRPH and other tests of liver function or some routine laboratory tests. SIRPH may reflect some hitherto unknown of unmeasured process in the diseased hepatic cells.


Arterial and coronary sinus catecholamines and cyclic-AMP during dynamic supine exercise in patients with chest pain. Arterial and coronary sinus catecholamine concentrations were measured during dynamic exercise in patients to assess the sympathetic response. Arterial concentrations increased from 1.77 nmol/1 (SEM = 0.53, n = 7) (control) to 2.95 nmol/1 (SEM = 0.65, n = 7) during exercise (0.05 greater than P greater than 0.01) and coronary sinus concentrations from 2.78 nmol/1 (SEM = 0.53, n = 7) (control) to 4.43 nmol/1 (SEM = 0.71, n = 7) (0.05 greater than P greater than 0.01). Resting, and exercise, arterial-coronary sinus differences in catecholamine concentrations were not statistically significant. In some patients, higher catecholamine concentrations occurred post-exercise than during exercise. The coronary sinus-arterial difference in catecholamine concentration during exercise was greatest in the one patient who developed angina pectoris. Cyclic-AMP concentrations were also measured, but these did not change significantly, consistent with the predominantly noradrenaline response to exercise.


Effect of a human serum thymic factor on hydrocortisone-treated thymocytes. A human serum thymic factors (SF) stimulated cyclic adenosine-3' ,5'-monophosphate (cAMP) synthesis in normal mouse thymocytes. Such stimulation was no longer observed when thymocytes were depleted of hydrocortisone (HC)-sensitive cells. It was concluded that SF selectively stimulate cAMP in HC-sensitive cells. Furthermore, incubation of thymocytes with SF enlarged the population of HC-resistant thymocytes. These results suggest that SF might act on HC-sensitive thymocytes increasing their cellular cAMP level and inducing their transformation in HC-resistant cells.


The opiate receptor binding interactions of 3H-methionine enkephalin, an opioid peptide. 3H-Methionine enkephalin binds stereospecifically with high affinity to opiate receptors in rat brain membranes. Equilibrium experiments indicate two distinct dissociation constants with KD values of 1.8 and 5.8 nM respectively. 3H-Methionine enkephalin associates and dissociates from the opiate receptor with 8--10 fold slower kinetics than 3H-opiates. Though several opiates have similar affinities for sites labeled by 3H-methionine enkephalin, 3H-dihydromorphine and 3H-naloxone, some opiates such as morphine, dihydromorphine and oxymorphone are only one tenth as potent in competing for 3H-methionine enkephalin as for 3H-dihydromorphine and 3H-naloxone binding. As with other opiate agonists, 5--10 mM sodium selectively decreases the binding of 3H-methionine enkephalin. At 26 degrees C, 0.1--1.0 mM manganese but not magnesium or calcium increases the binding of 3H-methionine enkephalin, while at 0 degrees C manganese decreases the binding of methionine enkephalin.


Behavioural despair in rats: a new model sensitive to antidepressant treatments. Rats when forced to swim in a cylinder from which they cannot escape will, after an initial period of vigorous activity, adopt a  characteristic immobile posture which can be readily identified. Immobility was reduced by various clinically effective antidepressant drugs at doses which otherwise decreased spontaneous motor activity in an open field. Antidepressants could thus be distinguished from psychostimulants which decreased immobility at doses which increased general activity. Anxiolytic compounds did not affect immobility whereas major tranquilisers enhanced it. Immobility was also reduced by electroconvulsive shock, REM sleep deprivation and "enrichment" of the environment. It was concluded that immobility reflects a state of lowered mood in the rat which is selectively sensitive to antidepressant treatments. Positive findings with atypical antidepressant drugs such as iprindole and mianserin suggest that the method may be capable of discovering new antidepressants hitherto undetectable with classical pharmacological tests.


Neurochemical and behavioral effects of clonidine and related imidazolines: interaction with alpha-adrenoceptors. Clonidine and other imidazolines, including phentolamine, inhibited the norepinephrine-elicited accumulation of cyclic AMP in brain slices via blockade or postsynpatic alpha-adrenoceptors. Clonidine (0.0125--0.050 mg/kg, i.p.) antagonized the increase in locomotor activity induced by amphetamine in a dose-dependent manner. It is suggested that the common ability of clonidine and phentolamine to antagonize norepinephrine-stimulated accumulation of cyclic AMP provides a possible explanation for the similar behavioral effects of these compounds, including their antagonism of the locomotor effects of amphetamine.


An assessment of the effect of hormone and neurotransmitters on adenine and guanine derivatives simultaneously in rat brain cortical slices. A A 2-dimensional thin-layer method has been developed for the separation on cellulose of adenine and guanine derivatives. Using incubated rat cerebral cortex slices it was shown that noradrenaline and acetylcholine stimulated cAMP and cGMP production respectively but glutamate and gamma-aminobutyric acid stimulated production of both cyclic nucleotides.


Neutrophils are mediators of antiviral immunity. The paper presents evidence that polymorphonuclear neutrophils upon stimulation with herpesvirus-induced antigens release a material inhibitory to virus infection. The material does not appear to be identical to type I or II interferon.


Effect of carbenoxolone on phosphodiesterase and prostaglandin synthetase activities. Carbenoxolone inhibited in vitro cAMP and cGMP phosphodiesterases in a concentration-dependent and noncompetitive manner. Prostaglandin synthetase activity of rabbit kidney medulla was slightly stimulated by carbenoxolone 0.1--0.5 mM, but inhibited by higher concentrations.


Ergoline derivatives with oral, prolonged, alpha-adrenolytic activity. The alpha-adrenolytic activity of 1-methyl-10-methoxydihydrolysergol 2-(3,5-dimethyl)pyrrolcarboxylate (XV, formula II) is, both in vitro and parenterally, quite similar to that of dihydroergotamine. By oral route the new compound is more active, and longer lasting than dihydroergotamine.


[Research on substances with antiviral activity. VIII. Benzindanoncarboxylic acids]. New derivatives (esters, amides, nitriles, amidines, thiosemicarbazones and guanylhydrazones) of 1H-benz[e]indene-2,3-dihydro-5-methyl-1-oxo-3-carboxylic acid and 1H-benz[e]indene-2,3-dihydro-7-methyl-3-oxo-1-carboxylic acid were prepared and tested for antiviral activities in vitro against vaccinia virus, HID stock and parainfluenza type 3 virus, HA-I/CR-8 stock. Four compounds (II g), (II i), (III b), (III g), showed weak activity in vitro against vaccinia virus.


Peripheral nerves in early experimental diabetes: expansion of the endoneurial space as a cause of increased water content. The aim of the present study was to examine whether the nerve water content and the Schwann cell cytoplasm are increased in early experimental diabetics, as suggested in the sorbitol theory. The sciatic nerves of streptozotocin diabetic rats were found to have an increased wet weight. The amount of Schwann cell cytoplasm was reduced by 30%. The increased wet weight was paralleled by enlargement of the cross sectional area of the nerve which was explained by an expansion of the endoneurial space. The findings indicate the existence of endoneurial oedema and are in part in conflict with the sorbitol theory. Extension of the space surrounding the nerve fibres may explain the increased resistance to ischaemia in diabetic patients.


Activation of human adenylate cyclase in the upper gastrointestinal tract by vasoactive intestinal polypeptide. The effects of various polypeptide hormones known to inhibit gastric acid secretion were tested on the adenylate cyclase system in human gastric and duodenal mucosal homogenates. Glucagon and secretin failed to stimulate the enzyme system in the stomach. The latter hormone produced a small but significant activation of the duodenal cyclase. The vasoactive intestinal polypeptide (VIP), however, induced a dose-dependent increase of enzyme activity throughout the stomach and the duodenum. Maximal effects (1.8 to 3.0-fold increase) were observed at a VIP-concentration of about 10 microgram per ml. Because the entire physiological role of VIP in gastric function has not been defined, ipt cannot be discerned whether the VIP-stimulated adenylate cyclase is linked to inhibition of gastric acid secretion or to another as yet unrecognized effect of this hormone in human gastric function.


Abrogation of BCG-contact induced tumour inhibition by silica: implications for the mechanism of action. The outgrowth of tumours from inocula of syngeneic rat tumour cells injected in admixture with BCG (Glaxo strain) is suppressed, the tumour cells apparently being destroyed in the milieu of the granulomatous reaction to the mycobacteria. The possible candidacy of macrophages as the major "effector cell" component was examined in experiments conducted in vivo using silica, a selective macrophage toxin. Intraperitoneal (i.p.) administration of this agent abolished contact-induced inhibition of BCG against three transplanted rat sarcomas, although for two of them the abrogatory effect could not be achieved without prior in vitro cultivation of the tumours. It is suggested that i.p. silica not only destroys macrophages within the cavity but accomplishes systemic depletion to the extent that granulomata contain insufficient macrophages for tumour rejection. This deficit may be compensated to a variable extent by the presence of host macrophages within the initial bacterial: tumour cell inoculum, but in the absence of these cells, total abrogation of the inhibitory effect of BCG is the invariable outcome in silica-treated hosts.


Inhibition of VIP-stimulated intestinal secretion and cyclic AMP production by somatostatin in the rat. The effect of somatostatin on colonic secretion induced by 10(-8) M vasoactive intestinal peptide (VIP), 10(-2) M theophylline, and 2 X 10(-3) M dibutyryl cyclic AMP was studied in muscle-stripped everted open rat colon sacs. The secretory response to VIP, measured as the decrease in net absorptive flow rate (microliters 30 min-1 mg-1 of dry weight), was maximal and equalled the responses to theophylline or dibutyryl cyclic AMP. Somatostatin (10(-5) M) blocked completely the secretory response to VIP but only partially the secretory response to theophylline or dibutyryl cyclic AMP. This difference in the extent of inhibition suggested that somatostatin exerted an inhibitory effect both before and after the point of generation of intracellular cyclic AMP. In order to test the hypothesis that one component of the action of somatostatin involved inhibition of the production of cyclic AMP, measurements of this nucleotide were made in isolated rat colon cells. Control levels of cyclic AMP measured by radioimmunoassay (12.6 +/- 1.6 pmoles per 10(6) cells) were not affected by 10(-5) M somatostatin. VIP (5 X 10(-8) M) increased cyclic AMP levels 2-fold (P less than 0.01) and this increase was blocked by somatostatin. The results indicated that somatostatin inhibits colonic secretion by exerting effects at two sites: one site lies at, and another beyond, the point of generation of intracellular cyclic AMP.


Mechanism of action of C. parvum on a solid, subcutaneous mouse tumour. The effects of i.v. C. parvum on the growth of a.s.c. inoculum of the Lewis lung carcinoma were studied in normal C57 B1 mice, and in those in which separate components of the immune response were impaired. C. parvum given either at the same time as tumour inoculation or when the carcinoma was 1 cm in diameter, and fully vascularised, reduced tumour growth. Macrophages were impaired by silica (Si), cortisone acetate (CA) or trypan blue (TB), and T cells by thymectomy and sublethal irradiation (TXR) or antilymphocyte serum (ALS). TB did not affect tumour size, whereas Si, CA, TXR, or ALS (initiated before tumour inoculation) reduced it. When C. parvum was given in combination with TB, or ALS (initiated after tumour inoculation), its antitumour effects were unchanged. C. parvum given after ALS (before tumour), TXR, Si, or CA had no further effect on tumour growth. We have shown that C. parvum inhibits the growth of inoculated Lewis tumour. However, it has not been possible to clearly define in vivo the immunological mechanisms involved.


Vagal stimulation and its role in eliciting gastrin but not glucagon release from the isolated perfused dog stomach. Electrical stimulation (10 V, 10 Hz, 3 min) of both dorsal and ventral vagal trunks of the isolated canine stomach perfused with whole blood induced strong gastric contractions, transient release of cyclic GMP and marked release of gastrin. No gastric-glucagon release was elicited either at 'normal' (4.8 +/- 0.1 mmol/l) or at low (1.5 +/- 0.1 mmol/l) concentrations of blood glucose. It is concluded that, in conditions effective for the stimulation of gastrin release, electrical stimulation of the vagus nerves does not stimulate glucagon release from the isolated perfused dog stomach. Thus one of the well-accepted mechanisms controlling pancreatic-glucagon secretion, vagal stimulation, is ineffective on gastric-glucagon release.


Plasma cyclic AMP levels during a secretin-caerulein pancreatic function test in liver and pancreatic disease. Plasma cyclic AMP levels were determined during a 40 minute secretin infusion (1 Cl.U kg-1h-1) followed by a 40 minute combined secretin (1 Cl.U kg-1h-1) caerulein (75 ng kg-1h-1) infusion. In nine healthy subjects, both secretin alone and secretin in combination with caerulein did not affect plasma cyclic AMP levels. The same was observed in six patients with chronic pancreatitis. By contrast, in patients suffering from liver disease (nine cases) or extrahepatic cholestasis (six cases), secretin elicited large increases in plasma cyclic AMP concentration; the mean values attained being, respectively, seven and four times higher than before the infusion. On the other hand, increases in plasma cyclic AMP 10 minutes after a bolus injection of glucagon (1 mg) were four times lower in the liver disease group as compared to the controls. The results reported here suggest that the liver plays a major role in the degradation of plasma cyclic AMP produced by target tissues responding to secretin, and in the release of cyclic AMP under glucagon. Liver disease reduce the capacity of the liver to clear cyclic AMP from the blood. The pancreas does not contribute significantly to the cyclic AMP in the blood.


Lymphoid cell lines: in vitro cell markers in correlation to tumorigenicity in nude mice. Tumorigenicity of lymphoid cell lines of different origin upon xenotransplantation (s.c. and i.m.) in cell concentrations of 1 X 10(6) cell/inoculum into nude mice was correlated to karyotype, presence of the 14 q + marker, EBV reactivity and immunological markers. Lymphoblastoid cell lines (LCL), lacking the 14 q + marker failed to produce tumors independent upon diploidy or aneuploidy. Lymphoma-type lines of Burkitt lymphoma, lymphosarcoma and Hodgkins disease-origin, genotypically aneuploid, and expressing the 14 q + marker were tumorogenic in nude mice, when inoculated in the same cell quantities where LCL failed to form tumors. Tumorgrowth in nude mice was independent upon the presence of EBV in the inoculated cells.


[Acupuncture as after care following injuries of the upper extremities]. Acupuncture is able to improve restriction of motion in general and in the smaller joints of the upper extremity, disturbances in circulation, posttraumatic swelling, reversible nerve lesions and ulcers following burns or pressure of a plaster. Localisation of the points for acupuncture in the different conditions and the possibilities of their combinations are described. Acupuncture is performed bilaterally at symmetrical points, unilaterally in pareses, contralaterally in amputees and in cases with dressings. During ear acupuncture the analgesic effect occurs more quickly, but has a shorter duration of action; in paralyses it was without success. The course of the treatment was controlled by clinical examination and measurement of the range of motion.


[Embryology and etiology of the Poland and Amazone syndromes]. In 1841 A. POLAND described a rare complex of malformations in the male, characterized a unilateral pectoral muscle defect combined with ipsilateral symbrachydaktyly. If in the female a POLAND Syndrome is associated with ipsilateral hypo- or aplasia of the breast as a leading feature, we propose to name this entity "Amazone Syndrome". Altogether 5 POLAND Syndromes and 40 "Amazone Syndromes" were seen in our department. For the latter a morphological grading into four groups was performed for clinical purposes. The malformation complex of the poland syndrome develops between the 2nd and 5th week of gestation as a result of damage to an upper extremity bud. It is from this extremity-"Anlage" that the muscles of the shoulder girdle and the lateral thoracic wall (with common innervation by the brachial plexus) arise. In early embryonal life the lacteal ridge lies in close proximity to the exrremity bud. The "Amazone Syndrome" can be explained by simultaneous damage to an extremity bud and the lacteal ridge. Its aetiology is not clear. Lack of embryonal material, heredity, and teratogenic damage by medication are discussed. In our patients there was a strong suspicion of unwanted pregnancies with failed induced interruptions. On the other hand some siblings had various other congenital deformities. In conclusion we may state that any inhibition of the normal growth of the upper extremity bud may lead to the POLAND Syndrome, while a simultaneous damage to the extremity bud and the close-by lacteal mound may result in the "Amazone Syndrome.


[Repeated contrast cisternography after previous use of an oily contrast medium (author's transl)]. Twenty-four control cisternograms were carried out at two to 22 months after lumbar injection of 1.0 to 1.5 ml. Duroliopaque; 20 of these were successfully performed without a fresh lumbar puncture. In most cases, the amount of contrast medium available was less than at the original examination, but was sufficient for definite exclusion of a space-occupying lesion. In four cases only the contrast medium could not be mobilised or manoeuvred into the cisterns and a fresh lumbar puncture was required for further contrast injection.


[In-flow of contrast into the liver--a rare complication during lymphography (author's transl)]. A lymphogram is described in which Lipiodol was found to have reached the liver from pelvic lymphatics. The common iliac and para-aortic lymph nodes had been invaded by tumour from a squamous carcinoma and this had produced complete lymphatic obstruction of the normal lymph paths. In addition, there was occlusion of the bifurcation of the iliac veins due to tumour, a finding also described by other authors. The causes for the finding of Lipiodol in the liver are discussed. The pathway by which the Lipiodol had reached the liver from the pelvic lymphatics, and the reaction of the liver parenchyma to the contrast medium, are considered.


[The use of computer tomography for radiation therapy of cerebral tumours (author's transl)]. The application of computer tomography should be extended for indicating the need and choice of radiotherapy for cerebral tumours, and for observing the effects of treatment. In 26 patients it has been shown that CT is superior to conventional neuro-radiological methods in showing indications and in planning of radiation therapy. Forms of treatment can be suitably adapted where the histological type of the tumour is known. In most cerebral tumours, the beneficial effect of irradiation has been evident in our patients. Our criteria were based on the concept of "clinical malignancy", no attempt was made at a histological classification of the tumours.


[Cytodiagnosis of the radiologically "occult" and the "in situ" lung neoplasm]. This paper reports on six patients, in whom a so-called "carcinoma in situ" or a radiologically "occult" carcinoma of the lung was diagnosed by means of cytological sputum examinations. On the basis of these cases, the problem of early localization of lung carcinomas is discussed. The cytological diagnosis of an occult lung carcinoma is practically always the result of an incidental sputum examination. Nevertheless, the value of "mass screenings" is doubtful, whereas systematic sputum examinations of so-called high risk patients seem so much the more indicated.


[Aspects of long-term therapy using long-term neuroleptics -- experiences and viewpoints related to practice]. Practical aspects of long-term neuroleptic therapy are discussed in the light of the author's own experience and observations; reference is made to the relevant literature. The author looks at the terms "sociogenic" and "somatogenic" in the context of treatment of schizophrenia and finds them to be not so antithetic as has traditionally been supposed. He describes the advantages and disadvantages of long-term neuroleptic therapy, both oral and injectable. He analyses the starting of a rational long-term therapy in the light of various factors. He examines the effective spectra of the traditional long-term neuroleptica and finds little justification for the different specific indications claimed for the various drugs. On the example of selected patients suitable for this therapy he considers the causes of the disorders and the primary personality and early development of the subjects, discussing the nature of the illness from the nosological and clinical point of view, the social and economic situations, the type of follow-up, and the physician-patient relationship. Emphasis is laid on the significance of side-effects and of individual reactions to long-term neuroleptics. The questions how, when and where a patient should be put onto long-term medication are treated, with a critical comparison of high and traditional doses, and a recommendation that duration and stoppage of therapy be decided on the basis of individual case factors. Special attention is paid to the so-called pharmacogenic depressions. Other indications of long-term neuroleptic therapy are mentioned. The author considers the integration of pharmaco-therapy into a complex rehabilitation program to be essential; without this there can be no progress in the treatment of schizophrenia.


[Clinicopathological study of diffuse carcinoma of stomach (author's transl)]. The biological behavior of ulcer type gastric carcinoma was studied on 114 cases of diffuse carcinoma (Borrmann's 4 type) and 262 cases of early like advanced carcinoma (including superficial spreading type). In both types of gastric carcinoma, the age distribution, location, ulcer with cancer focus and prognosis differed greatly. The early like carcinoma was speculated to have advanced maintaining the groos findings of early gastric carcinoma, and its location and associated ulcer were the same as the early ulcer type of carcinoma. The prognosis of this type of carcinoma was good, showing a figure of 70% in 3 year survival rate. On the other hand, diffuse carcinoma demonstrated diffuse extensive infiltration of tumor cells along the gastric wall, resulting in poor prognosis with a 3 year survival rate of almost 0%. Histologically, diffuse type of carcinoma showed lymphatic infiltration of tumor cells, and this is probably the main reason for the diffuse infiltration in this type of carcinoma. Diffuse carcinoma is, therefore, considered to be one special type of carcinoma having different biological behavior compared with the other ulcer type of carcinoma, and diffuse carcinoma is not the terminal stage of early like advanced carcinoma. There are three stages in diffuse carcinoma: 1. Infiltrative stage: wide spread infiltration of cancer cells through lymphatic channels (lymphangiosis carcinomatosa) 2. Edematous stage: soluble collagen appearing in gastric wall 3. Sclerosing stage: soluble collagen changing into insoluble collagen leading to marked thickening and stiffness of the gastric wall. This is the end stage of gastric diffuse carcinoma. It is difficult to explain that the marked fibrosis of gastric wall is a result to stromal reaction from tumor cell infiltration, since extensive fibrosis is found in areas without tumor cells and stiffness of the gastric wall occurs in a too short period of time. The production of abundunt soluble collagen is probably related to cancer cells.


Urinary and plasma cyclic AMP levels during short term starvation in obese man: response to glucagon stimulation. Glucagon is known to elevate the intracellular concentration of cyclic AMP in the hepatocyte. The increase in intracellular cyclic AMP is reflected by an increase in the plasma concentration of the nucleotide. Intravenous glucagon stimulation was performed on six obese non-diabetic human subjects before and after a three day fast. All patients responded to starvation by a lowering of plasma immunoreactive insulin and blood glucose. Whereas the plasma immunoreactive glucagon concentration increased over the three day period, the plasma and urinary cyclic AMP did not significantly change. Intravenous glucagon promoted qualitatively similar increases in the blood glucose and plasma concentrations of insulin and cyclic AMP before and after three days starvation.


Effect of treadmill exercise on plasma and urinary cyclic adenosine 3'5'-monophosphate. The effect of treadmill exercise on plasma and urinary cyclic adenosine 3'5' monophosphate levels (cyclic AMP) was studied in twelve healthy subjects. Plasma cyclic AMP levels were found to be markedly elevated without significant changes in urinary cyclic AMP or cyclic AMP/creatine ratio. Most likely altered plasma glucagon and catecholamine levels were responsible for these changes.


Plasma renin activity and urinary aldosterone in Cushing's syndrome. The renin-angiotensin-aldosterone system has been evaluated in 19 patients with Cushing's syndrome due to bilateral adrenal hyperplasia and in 2 patients with unilateral adenoma. In the first group urinary aldosterone was within the normal limits with a mean of 8.3 +/- 1.86 microgram/24 h. Aldosterone excretion did not change significantly after furosemide administration, ACTH infusion or dexamethasone. Upright PRA was suppressed in 9/16 patients with a mean of 4.9 +/- 1.85 ng/ml/3 h and showed only a slight response to furosemide. Dexamethasone alone did not produce any change. Both aldosterone and PRA were to some extent stimulated by an association of dexamethasone and furosemide. In the 2 patients with adenoma, aldosterone excretion was also normal, but PRA was very elevated. From our data it is concluded that in Cushing's syndrome due to bilateral hyperplasia, PRA and aldosterone excretion are partially suppressed. From our results on plasma deoxycorticosterone and corticosterone concentration it seems unlikely that these mineralocorticoids are the major cause of this phenomenon. However, it may not be excluded that other yet unidentified hormones could play some role in the pathogenesis of hypertension and renin suppression in Cushing's syndrome.


A program for continuing care: implementation and outcome. The authors used the records of the continuing-care program at a large public hospital in Los Angeles to determine the success of referrals for aftercare. A total of 552 patients discharged during an eight-month period were referred for continuing care; 418 of the patients kept their first appointment. Patients were most likely to keep their first appointment if it was set up for within three days of release from the hospital and if it was confirmed before discharge from the hospital. If an appointment could not be arranged within three days, a phone call to the patient improved the referral completion rate.


Tumor immunity: relevance of animal models to man. The conclusions that can be drawn from results of animal experiments often provide important insights into the mechanisms of human disease and the rationale for effective therapy. A few years ago, animal models of tumor immunity revealed that both specific and nonimmune specific stimulation of the immune system could effectively prevent or cure cancer. Extensive clinical studies have not convincingly demonstrated that the results with the animal models used are reproducible in humans. New animal models more closely reflecting the properties of human cancer are needed to determine the applicability of cancer immunotherapy to man.


The importance of the ratio between effector and target cells for detection of serum blocking of tumor lymphocytolysis. Sera from tumor-bearong guinea pigs were tested for their capacity to influence the magnitude of lymphocytolysis of tumor cells in vitro as measured by 51Cr release. Thirty-three sera were tested in 124 experiments in which the effector to target cell ratio (E:T) was 200:1. Significant blocking of cell-mediated lysis occurred in 62% of the experiments, whereas significant potentiation occurred in 7%. Six sera were found which showed significant blocking in some experiments and significant potentiation in other experiments. One of these sera was further evaluated by varying E:T, using lymphocytes from three immune donors. For each of the three populations of immune lymphocytes, significant blocking was demostrable only at one of three E:T tested. Blocking was observed only with an E:T of 200:1 in two cases. In a third case, blocking was observed at an E:T of 50:1, whereas an E:T of 200:1 gave rise to significant potentiation. The effect of these sera on lymphocytolysis in vitro is clearly dependent upon the E:T ratio.