[Influence of active vitamine D3 on bones (author's transl)].

Influence of 1 alpha-OH-D3, an active vitamine D supplement, on bone cells was studied, histologically and electro-microscopically observing the vitamine D objective bone cell after treatment with the drug in a dose of 50 microgram/kg/day. Light-microscopic findings revealed that osteogenesis gradually became dominant after transient osteoporosis, leading to a disproportional state of the bone remodelling. In electromicroscopy, no morphological changes were observed, though the number of osteoclasts increased five days after treatment. Hypofunctional findings, however, appeared seven and ten days after treatment. On the other hand, findings of distinguishable osteogenesis, e.g. well-developed golgi complex (G), endoplasmic reticulum (ER), mitochondria (M), were observed in the osteoblast and osteocyte, defining the strong influence of vitamine D in a large dose on the osteoblast-osteocyte system. Especially, the osteoblast seemed to be influenced earlier and to a greater degree. We concluded that 1 alpha-OH-D3 in doses of 0.02 and 0.08 microgram/kg/day hardly caused any morphological changes in the bone cell in our experiment.


Detection by proton nuclear magnetic resonance of elevated lactate concentration in serums from patients with malignant tumors.

Proton nuclear magnetic resonance (1H-NMR) spectra of the serum specimens from patients with malignant tumors were compared with those from presumably healthy persons. We found that 87% of serum specimens from the patients yielded a common proton signal, which was ascribed to the methyl protons of lactic acid; whereas only 9% of serum specimens from the healthy persons tested gave this signal. On the basis of these results we concluded that the lactate level in the serum can be used as a criterion for the diagnosis of cancer in humans and that the determination of lactate concentration in the serum is easily performed by means of 1H-NMR.


Preoperative estimation of operative risk in liver surgery, with special reference to functional reserve of the remnant liver following major hepatic resection.

Hepatic functional reserve was evaluated in 76 patients with known liver, biliary tract or pancreas diseases using kinetic analysis of removal of indocyanine green (ICG) with special reference to maximal removal rate (Rmax). In surgery other than hepatectomy, if ICG Rmax is below 0.4 mg/kg/min the operative risk should be considered high. In hepatic surgery, even if ICG Rmax is above 0.4 mg/kg/min the operative risk was high and it should be required to be above 1.0 mg/kg/min for extended lobectomy or hepatectomy. Furthermore, prior to hepatectomy the functional reserve of the remnant liver was estimated from an effective liver volume rate, calculated from the rate of uptake of radioisotope as measured by on-line computer system, and ICG Rmax. The functional reserve of the remnant liver was compared with the operative results, such as morbidity or mortality for each patient. When ICG Rmax of the remnant liver was below 0.4 mg/kg/min the prognosis was poor in hepatic surgery, because of there were three postoperative death and seven complications such as ascites or liver failure in the ten cases. On the other hand, when above 0.4 mg/kg/min, the prognosis was good without any postoperative death or complications in the twelve cases of hepatic surgery, as the completely same results for general surgery, in which it is as well when ICG Rmax of whole liver is above 0.4 mg/kg/min.


Isolation and immunological characterization of an iron-regulated, transformation-sensitive cell surface protein of normal rat kidney cells.

We have analyzed the surfacr proteins of cultured normal rat kidney (NRK) cells and virus-transformed NRK cells subjected to iron deprivation. Such a treatment specifically induces two transformation-sensitive plasma membrane-associated glycoproteins with a subunit molecular weight of 160,000 (160 K) and 130,000 (130 K) daltons in NRK cells. In these cells the 160 K glycoprotein is readily available to lactoperoxidase-mediated iodination, and the 130 K is apparently inaccessible to iodination. Major differences were revealed when iodinated membrane proteins of normal and virus-transformed cells subjected to iron deprivation were compared. In Kirsten sarcoma virus-transformed NRK cells the 160 K glycoprotein was weakly labeled. In two clones of simian virus 40-transformed NRK cells the 160 K glycoprotein was weakly labeled or not at all. The 130 K glycoprotein was inaccessible to iodination in all virus-transformed cell lines. The 160 K and 130 K glycoproteins were isolated from plasma membranes of NRK cells using preparative SDS gel electrophoresis. Antibodies generated against these glycoproteins stained the external surfaces of NRK cells and induced antigen redistribution. Evidence presented suggests that 160 K and 130 K are plasma membrane-associated procollagen molecules. A possible interaction of these proteins with transferrin is also described. The data suggest that these proteins may have an important role in the sequence of events leading to transformation.


Fibronectin and proteoglycans as determinants of cell-substratum adhesion.

When normal or SV40-transformed Balb/c 3T3 cells are treated with the Ca++-specific chelator EGTA, they round up and pull away from their footpad adhesion sites to the serum-coated tissue culture substrate, as shown by scanning electron microscope studies. Elastic membranous retraction fibers break upon culture agitation, leaving adhesion sites as substrate-attached material (SAM) (Cells leave "footprints" of substrate adhesion sites during movement by a very similar process.) SAM contains 1-2% of the cell's total protein and phospholipid content and 5-10% of its glucosamine-radiolabeled polysaccharide, most of which is glycosaminoglycan (GAG). By one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis, there is considerable enrichment in SAM for specific GAGs; for the glycoprotein fibronectin; and for the cytoskeletal proteins actin, myosin, and the subunit protein of the 10 nm-diameter filaments. Fibrillar fibronectin of cellular origin and substratum-bound fibronectin of serum origin (cold-insoluble globulin, CIg) have been visualized by immunofluorescence microscopy. The GAG composition in SAM has been examined under different cellular growth and attachment conditions. Heparan sulfate content correlates with glycopeptide content (derived from glycoprotein). Newly attaching cells deposit SAM with principally heparan sulfate and fibronectin and little of the other GAGs. Hyaluronate and chrondroitin proteoglycans are coordinately deposited in SAM as cells begin spreading and movement over the substrate. Cells attaching to serum-coated or CIg-coated substrates deposited SAM with identical compositions. The proteoglycan nature of the GAGs in SAM has been examined, as well as the ability of proteoglycans to form two classes of reversibly dissociable "supramolecular complexes" - one class with heparan sulfate and glycopeptide-containing material and the second with hyaluronate-chondroitin complexes. Enzymatic digestion of "intact" SAM with trypsin or testicular hyaluronidase indicates that (1) only a small portion of long-term radiolabeled fibronectin and cyto-skeletal protein is bound to the substrate via hyaluronate or chondroitin classes of GAG; (2) most of the fibronectin, cytoskeletal protein and heparan sulfate coordinately resist solubilization; and (3) newly synthesized fibronectin, which is metabolically labile in SAM, is linked to SAM by hyaluronate- and/or chondroitin-dependent binding. All of our studies indicate that heparan sulfate is a direct mediator of adhesion of cells to the substrate, possibly by binding to both cell-surface fibronectin and substrate-bound CIg in the serum coating; hyaluronate-chondroitin complexes in SAM appear to be most important in motility of cells by binding and labilizing fibronectin at the periphery of footpad adhesions, with subsequent cytoskeletal disorganization.


Cholesterol levels and plasma membrane fluidity in 3T3 and SV101-3T3 cells.

Polyene antibiotics such as filipin selectively inhibit wheat germ agglutinin-induced agglutination of transformed and malignant cells compared to normal cells (Hatten ME, Burger MM: Biochemistry 18: 739, 1979). Since filipin binds specifically to cholesterol, we measured cholesterol levels in 3T3 cells and SV101-3T3 cells. SV101-3T3 cells contained 50-100% more cholesterol per cell than 3T3 cells. Both cell types were starved for cholesterol by growth in lipid-depleted medium plus 25-hydroxycholesterol. The cholesterol level of SV101-3T3 cells decreased by 30-50%, while the level in 3T3 cells remained constant. Filipin-stained SV101-3T3 cells revealed bright patches of filipin under fluorescence microscopy. These patches were absent in 3T3 cells and in SV101-3T3 and 3T3 cells starved for cholesterol. We selectively labeled plasma membranes of these cells with a spin label analog of phosphatidylcholine. The spin label indicated differences in plasma membrane fluidity that may be related to the different cholesterol levels in 3T3 and SV101-3T3 cells.


Tumorigenicity of revertant from an SV40-transformed line.

A syndrome of in vitro properties correlates with the tumorigenicity of SV40-transformed rodent cells. These properties are: plasminogen activator production, loss of large actin cables, and anchorage-independent growth. An established rat fibroblast line, its SV40 transformant, several T-antigen negative revertants, and a spontaneous retransformant isolated from one of the revertants were analyzed in vivo for their tumorigenicity and in vitro for the syndrome. The two transformed lines were highly tumorigenic, and had clearly abnormal in vitro properties. The parental rat line was weakly tumorigenic in nude mice and demonstrated a slightly transformed response in the in vitro assays. The revertants were completely nontumorigenic. Expression of the in vitro syndrome was not uniform for all revertants; however, most cell lines maintained the correlation of the syndrome and tumorigenicity.


Alteration of insulin binding and cytoskeletal organization in cultured fibroblasts by tertiary amine local anesthetics.

Tertiary amine local anesthetics cause a time- and dose-dependent, reversible increase in insulin binding sites in cultured chick embryo fibroblasts. Incubation of fibroblasts with 0.2 mM dibucaine for 3 h at 37 degrees C results in a twofold to threefold increase in insulin binding, with an increase in average number of binding sites (Ka = 3.0 X 10(7) M-1) from 9 X 10(3) per cell. Trypsin or ethyleneglycoltetraacetic acid (EGTA) alone increases insulin binding twofold to threefold, but fails to further increase 125I-insulin binding in cells pretreated with dibucaine. Transformation of chick embryo fibroblasts with Rous sarcoma virus causes a threefold to fivefold increase in insulin binding, which is not further increased by incubation with dibucaine. As demonstrated by transmission electron microscopy, dibucaine and trypsin also induce changes in the cytoskeleton of chick embryo fibroblasts, characterized by disorganization and disappearance of microfilament and microtubule bundles. These alterations are accompanied by gross morphologic changes, including rounding of cells and appearance of numerous ruffles and blebs on the cell surface. These observations are consistent with the hypothesis that expression of surface receptors in cultured chick embryo fibroblasts is related to the organization and disorganization of cytoskeletal structures.


Modulation of cell surface iron transferrin receptors by cellular density and state of activation.

This report describes investigations of plasma membrane transferring receptors on a variety of lymphoid cell lines and normal peripheral blood lymphocytes during activation and cell growth cycles. Transformed lymphoid cell lines have as many as 1,000 times the number of receptors found on normal resting lymphocytes. The number of iron transferrin receptors on continuous cell lines as well as normal human fibroblasts is down-regulated during the transition from log-phase growth to stationary plateau growth. When normal lymphocytes are transformed by mixed lymphocyte culture or mitogens, they rapidly express a 50-fold increase in the number of transferrin binding sites. This appearance of iron transferrin receptors anticipates nuclear changes during cell activation and subsequent mitosis of normal cells.


Polyoma T (tumor) antigen species in abortively and stably transformed cells.

Stable neoplastic transformation of cells by polyoma virus requires the particpation of two viral genes, designated ts-a and hr-t. The effects of mutations in these two genes on the patterns of T-antigen synthesis during productive infection have been previously described: ts-a mutants are affected in the "large" (100K) nuclear T antigen, and hr-t mutants are affected in the "middle" (36K, 56K, 63K) and "small" (22K) T antigens. The latter are associated predominantly with the plasma membrane (56K) and cytosol fractions, respectively. Here we examine the expression of the various forms of polyoma T antigen in nonproductive infection (abortive transformation) as well as in stably transformed cell lines of different species. The results on abortive transformation are essentially the same as those described above for productive infection. In stably transformed cells, the middle and small T antigens are seen to various extents. The large T antigen, however, is often absent or present below the level of detection. Clones lacking the large T antigen are found most often among mouse transformants, but are also seen among rat transformants. Retention of the 100K species in transformed cells therefore appears to be, at least in part, an inverse function of the level of permissivity of the host toward productive viral infection. These findings indicate that the induction of the transformed phenotype in both abortively and stably transformed cells generally does not require the large T antigen, but rather the products of the hr-t gene.


Effects of the tumor promoter TPA on the induction of DNA synthesis in normal and RSV-transformed rat fibroblasts.

Induction of DNA synthesis by the tumor promoter tetradecanoyl phorbol acetate (TPA) was studied in a line of cultured rat fibroblasts (Rat-1) and their Rous sarcoma virus-transformed derivative (Rat-1 (RSV)). Following serum deprivation for 54 h to achieve quiescence, semiconservative DNA replication was measured by incubation of cells in BrdUrd and FdUrd after serum stimulation in the presence or absence of TPA. Optimal concentrations of TPA (0.1--0.5 microgram/ml) in serum-free medium induced a small increase (10--15%) in the amount of DNA made over a 30-h period in both Rat-1 and Rat-1 (RSV) cells. When Rat-1 cells were stimulated by a 4-h serum pulse, 30% of the DNA was replicated by 30 h. If the serum pulse was followed by TPA addition, 70% DNA replication was observed. If the serum pulse was preceded by TPA addition, the onset of DNA synthesis was delayed by several houses, but stimulation of DNA synthesis occurred. In contrast, the Rat-1 (RSV) cells did not show an increased in DNA synthesis induced by TPA in similar protocols, but the serum-induced onset of DNA synthesis was delayed by several hours in the presence of TPA. Therefore, TPA acts as a co-inducer of DNA synthesis in the Rat-1 but not in the Rat-1 (RSV) cells. The parent alcohol, phorbol, was inactive in Rat-1 cells, but delayed the onset of DNA synthesis in the Rat-1 (RSV) cells. We conclude that the co-inducing and delaying activities of TPA on DNA synthesis appear to be distinct and to act a different points in the G1 phase of the cell cycle.


Structural and functional alterations in the surface of vascular endothelial cells associated with the formation of a confluent cell monolayer and with the withdrawal of fibroblast growth factor.

Vascular endothelial cells cultured in the presence of fibroblast growth factor (FGF) divide actively when seeded at low or clonal cell densities and upon reaching confluence adopt a morphologic appearance and differentiated properties similar to those of the vascular endothelium in vivo. In this review, we present some of our recent observations regarding the characteristics (both structural and functional) of these endothelial cells and the role of FGF in controlling their proliferation and normal differentiation. At confluence the endothelial cells form a monolayer of closely apposed and nondividing cells that have a nonthrombogenic apical surface and can no longer internalize bound ligands such as low-density lipoprotein (LDL). The adoption of these properties is correlated and possibly causally related to changes in the cell surface such as the appearance of a 60,000 molecular weight protein (CSP-60); the disappearance of fibronectin from the apical cell surface and its concomitant accumulation in the basal lamina; and a restriction of the lateral mobility of various cell surface receptor sites. In contrast, endothelial cells that are maintained in the absence of FGF undergo within three passages alterations that are incompatible with their in vivo morphologic appearance and physiologic behavior. They grow at confluence on top of each other and hence can no longer adopt both the structural (CSP-60, cell surface polarity) and functional (barrier function, nonthrombogenicity) attributes of differentiated endothelial cells. Since these characteristics can be reacquired in response to readdition of FGF, in addition to being a mitogen FGF may also be involved in controlling the differentiation and phenotypic expression of the vascular endothelium.


Regulation of dome formation in differentiated epithelial cell cultures.

Rat mammary (Rama 25) and dog kidney (MDCK) epithelial cell cultures formed 'domes' of cells due to fluid accumulation in focal regions between the culture dish and the cell monolayer. Addition of ouabain caused collapse of domes, suggesting that transport functions were required for maintenance of domes. Dome formation in both epithelial cell lines was stimulated by a broad spectrum of known inducers of erythroid differentiation in Fried erythroleukemia cells. Among these inducers were: 1) polar solvents such as dimethyl-sulfoxide, dimethylformamide, and hexamethylene bisacetamide; 2) purines such as hypoxanthine, inosine, and adenosine; 3) low-molecular-weight fatty acids such as n-butyrate; and 4) conditions expected to elevate levels of cyclic AMP. In the latter group were activators of adenylate cyclase such as cholera toxin and prostaglandin E 1; cyclic AMP phosphodiesterase inhibitors such as theophylline and 1-methyl-3-isobutylxanthine; and analogs of cyclic AMP. Induction of domes occurred 15--30 h after addition of inducer to the culture medium. Induction by chemicals was serum-dependent and required protein synthesis but not DNA synthesis. Induced dome formation was reversible after removal of inducer, requiring the continuous presence of inducer. Reversal was also observed after either either removal of serum or addition of inhibitors of protein synthesis. These results suggest that hypothesis that domes arise in these epithelial cultures by a process that is similar to cell differentiation and is influenced by cyclic AMP.


[Lysozyme activity in the saliva of industrial workers exposed to fluorine compounds].

The authors performed investigations in Phosphoric Fertilizers Works in Gdańsk to find out whether or not the work environment contaminated by fluorine compounds affects the activity of lysozome--the protective enzyme of saliva. In some in vitro experiments lysozome activity in exposed workers' saliva was found to be significantly lowered, as opposed to that in unexposed workers. In vivo experiments did not indicate any effects of work conditions on the activity of this enzyme. The obtained results, in vivo and in vitro, do not demonstrate any clear effects of work environment in the Phosphoric Fertilizers Works in Gdańsk, upon lysozome activity in workers' saliva.


Strain characteristics and features of ocular infection of herpes simplex virus type 1 isolates.

Herpes simplex virus type 1 isolates from 63 patients with herpetic keratoconjunctivitis were investigated. For the purpose of detecting strain differences neurovirulence was determined in mice, and viral DNA's were analyzed by digestion with EcoRI and Hind III restriction endonucleases. Neurovirulence differed by a factor up to 10(5) between individual strains and proved to be independent of the ratio between infectious and noninfectious particles in the stocks used. The endonuclease cleavage patterns revealed differences of the viral DNA structure which permitted us to distinguish seven clusters of strains. Correlations between neurovirulence and DNA markers could not be established nor could correlations be found between these markers and features of disease. The main reason for not finding a relationship in the latter case may be attributed to the significant role of host factors in the course of herpetic diseases.


Infection and persistence of varicella-zoster virus in lymphoblastoid Raji cell line.

Infection of Raji cells by varicella-zoster virus (VZV) resulted in permissive infection with establishment of a persistently infected lymphoblastoid cell line. VZV antigens of the membrane and nuclear type, as detected by the indirect immunofluorescence membrane antigen (IFAMA) and anticomplement immunofluorescence (ACIF) tests, were observed. Minute amounts of infectious virus were detected by co-cultivation of VZV-infected Raji cells (Raji-VZV), with permissive human embryo fibroblasts (HEF). The virus isolated was found to be similar to the parent strain. Transient induction of Epstein-Barr viral capsid antigen (EB-VCA) was also observed. The persistently infected Raji-VZV cell line, when free of EB-VCA, was found suitable for measuring antibodies to varicella-zoster virus. The possible interaction in the infected Raji cells between EBV, which is implicated in human malignancy, and VZV which belongs also to the herpes group of viruses, is discussed.


Muscle and nerve changes induced by perhexiline maleate in man and mice.

Perhexiline maleate therapy may produce a polyneuropathy as well as other side effects in man. The Schwann-cell origin of the neuropathy is demonstrated by morphologic studies. Ultrastructural studies of muscle and nerve biopsies showed the presence of intracytoplasmic inclusions, primarily in Schwann cells, muscle fibers, and endothelial cells. These inclusions were polymorphic and varied from one cell type to another. They were frequently associated with calcium deposits, and their structure suggested a lysosomal origin. It was possible experimentally to reproduce similar inclusions in mice. Furthermore, numerous tubular aggregates were observed in intoxicated mouse muscle fibers.


Cyclic 3',5'-adenosine monophosphate phosphodiesterase (cAMP PDE) and cyclic 3',5'-guanosine monophosphate phosphodiesterase (cGMP PDE) in microvessels isolated from bovine cortex.

It was established that microvessels of a bovine cortex exhibit significant cyclic 3',5'-adenosine monophosphate phosphodiesterase (cAMP PDE) and cyclic 3',5'-guanosine monophosphate phosphodiesterase (cGMP PDE) activities. These activities are dependent on the presence of Mg2+. Absence of Ca2+ was virtually without effect. When both Mg2+ and Ca2+ were absent, PDE activities increased compared with activities observed in the absence of Mg2+. Xanthines (caffeine, theobromine, and theophylline) were better inhibitors of cAMP PDE than of cGMP PDE. Imidazole, in very high concentration (1 X 10(-2) M) only, exhibited PDE stimulatory activity at high concentrations of both substrates. Otherwise, it exhibited PDE-inhibitory properties.


Animal models of alcoholic neuropathy: morphologic, electrophysiologic, and biochemical findings.

A chronic high alcohol intake was induced in rats through the use of two procedures: the schedule-induced polydipsia technique and the liquid diet technique. Rats consumed 11-12 g of ethanol per kilogram body weight per day for 16 to 18 weeks. Morphologic evidence of a mild distal axonal neuropathy in the ventral caudal nerve was proposed. The red blood cell transketolase levels were normal, indicating that the rats were not deficient in thiamine and suggesting that the axonal degeneration was due to the direct toxic effect of alcohol. Axonal transport studies demonstrated a significant increase in the amount of acetylcholinesterase transported in an orthograde direction in the sciatic nerves of alcohol-exposed rats, and indicated no change in the transport of choline acetyltransferase or in the specific binding of colchicine by neurotubulin.


Studies on the glycoprotein component of (Na+-k+) atpase from guinea pig brain.

In this study an attempt was made to elucidate the possible mechanism of the brain microsomal (Na+-K+)ATPase inhibition based on the assumption that glycoprotein part of the enzyme is exposed on the outer membrane surface. In our experiments the modification with concanavalin A of sugar end groups exposed by neuraminidase treatment resulted in a significant decrease of the brain (Na+-K+)ATPase activity. The percentage of the enzyme inhibition by concanavalin A binding to the neuraminidase-treated preparation corresponds to the amount of liberated sialic acids. The modification of the glycoprotein part of the brain (Na+-K+)ATPase complex by neuraminidase and concanavalin A treatments did not affect K+-nitrophenylphosphatase activity.


beta-Adrenergic receptors of frog erythrocytes. Biochemical sequelae following stimulation with isoproterenol.

Following persistent stimulation of beta-adrenergic receptors of frog erythrocytes with (-)-isoproterenol, the cyclic adenosine 3',5'-monophosphate-dependent protein kinase (cAMP-dependent protein kinase) (EC 2.7.1.37) was activated for several hours. This activation outlasted the duration of the increase of cAMP content. Following a persistant stimulation of beta-adrenergic receptors with isoproterenol, the phosphorylation of selective membrane proteins was increased. This increase in phosphorylation lasted longer than 4 hr but less than 12 hr. Between 2 and 4 hr after receptor stimulation the loss of beta-adrenergic receptor from plasma membrane was maximal, and the phosphorylation of two membrane proteins characterized by molecular weights of 60,000 and 38,000 daltons was selectively enhanced. In addition we found that isolated erythrocytes are capable of synthesizing RNA and polypeptides and that incubation with (-)-isoproterenol indices a long-term delayed increase of the synthesis of erythrocyte proteins. This increase in the synthesis of proteins appears to require new RNA synthesis. Thus the possibility can be entertained that this delayed increase in protein synthesis participates in the new synthesis of receptor and is operative in the termination of beta-adrenergic receptor subsensitivity elicited by a persistent stimulation with (-)-isoproterenol.


Phosphorus 31 NMR of neuroblastoma clonal lines. Effect of cell confluency state and dibutyryl cyclic AMP.

Phosphorus nuclear magnetic resonance is used to study changes in the levels of the major phosphate-containing intermediary metabolites concomitant with induced cellular differentiation in the N-18 and C-46 neuroblastoma clonal lines. The study reveals differences between the 31P-NMR profiles of the two clonal lines and also striking differences attendant to dibutyryl cAMP-mediated morphological differentiation in the N-18 clone. Phosphorus-31 NMR would appear to provide a new technique with which to study genetic differentiation.


Lacrimal gland and fossa lesions: an approach to diagnosis and management.

Lesions of the fossa of the lacrimal gland present special problems in diagnosis and management. Clinical recognition and differentiation of benign mixed cell lacrimal gland tumors from other inflammatory and neoplastic lesions of the lacrimal gland fossa are feasible and should provide the basis for the approach to proper management. Lesions conforming to the clinical picture of benign mixed cell lacrimal gland tumors need not undergo incisional biopsy, but require en bloc excision with adjacent tissues through a lateral orbitotomy to ensure complete excision and to prevent late recurrence. Incisional biopsy is indicated and should not be delayed in suspected infectious and noninfectious inflammatory lesions that have not responded rapidly to medical therapy, and in other lesions suspected of being neoplastic and not amenable to total local excision. An expedient approach to these lesions is essential if the prognosis of epithelial lacrimal gland neoplasms is to be improved.


[Angioimmunoblastic lymphadenopathy: ultrastructural study of lymph nodes. Possible relationship with Epstein-Barr Virus (EBV) (author's transl)].

Angioimmunoblastic lymphadenopathy (AIL) has been clinically and histopathologically described by several authors since 1972, but only a few papers have analyzed the morphological aspects of the lymph nodes as observed by using electron microscopy. In the past three years, we happened to see in the Curie Institute two typical cases of AIL, both of whose clinical and immunological data are briefly reported in this paper. We describe some peculiar patterns observed on the lymph nodes during the acute phase of the disease, by examination of semithin sections and electron-microscopic preparations. As have many others, we found strong diffuse B-lymphocyte proliferation, but we must emphasize that many plasmocytes had an obvious defect in cell maturation, but no monomorphous defect. We noted that the predominant postcapillary venules had hypertrophied endothelial and perithelial cells, and multilayering basal lamina. We found some unusual features such as lymphocyte and/or plasmocyte phagocytosis by macrophages, or the atypical aspect of eosinophilic granulations. Finally, in the cytoplasm of a few transformed lymphocytes we detected some viral-like particles resembling herpes virus. Whether this last finding may have been related to the pathogeny of AIL could not be resolved by using morphological techniques. In the second part of this paper, we discuss the etiology of the disease, taking in account our clinical, immunological, and morphological observations. The possible relation of AIL to infectious mononucleosis, and the hypothetical role of Epstein-Barr virus are discussed.


Psychologic correlates of serum indicators of stress in man: a longitudinal study.

Correlations between serum uric acid, cortisol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol and psychometric indices of stress--namely, anxiety, hostility, and depression--were investigated in 20 students over a 2 1/2 month academic quarter. There were three peak periods of stress, with two occurring during midterm and final exams. The observed changes in serum cortisol were highly correlated with changes in anxiety and depression. Serum uric acid was significantly depressed during the first half of the quarter, but returned to control levels after midterm exams. Serum cholesterol and the LDL cholesterol subfraction were significantly elevated above control levels and remained elevated throughout the quarter after the first day of classes. Absolute levels of HDL cholesterol varied little throughout the quarter, however, the ratio of HDL cholesterol/total cholesterol was significantly depressed twice in the quarter, each time following approximately 10 days after the peak period of stress.


Yielding ability of erect- and narrow-leaved rice mutant in heavy manuring and dense planting culture.

A dwarf rice mutant was induced through the chronic exposure of 60Co gamma-rays. This mutant was crossed with a heavy-panicle type variety, and semi-dwarf plants permitting a more efficient distribution of light within the canopy were obtained. They along with current commercial varieties and the parent mutant were assessed for yield components, the yielding ability at three levels of nitrogen fertilizer by varying plant density and several other agronomic traits. The ssmidwarf rice possessed significantly higher rates of net photosynthetic CO2 uptake compared with that of current commercial varieties, and it was improved in yield return by increasing nitrogen application. Manner of display of leaves could be manipulated by induced mutations.


Activity of human hippocampal formation and amygdala neurons during sleep.

Fine wire microelectrodes were implanted for diagnostic purposes in 17 patients suffering from psychomotor epilepsy. Single- and multiunit activity during waking and natural nocturnal slow wave sleep and REM sleep was recorded in the hippocampus (n = 42), hippocampal gyrus (n = 53), and amygdala (n = 32). The firing rates of hippocampal gyrus units usually decreased during slow wave sleep and then increased to levels equal to or above waking during REM. In contrast, the firing rates of hippocampal neurons generally increased during slow wave sleep and fell to very low levels during REM. The amygdala presented a more mixed pattern. Since the projection from the hippocampal gyrus to hippocampus is excitatory, their opposite patterns during sleep suggest that the tonic firing patterns of HC neurons may be mainly the result of subcortical afferents.


Opioid effects on computer-derived sleep and EEG parameters in nondependent human addicts.

After one adaptation night, intramuscular doses of methadone (7.5, 15, 30 mg/70 kg), morphine (10 or 20 mg/70 kg), or placebo were given to seven male nondependent opiate addicts at weekly intervals in a randomized cross-over design. After three adaptation nights, heroin (3, 6, 12 mg/70 kg) was compared with morphine and placebo by means of a similar design in seven other subjects. Using electroencephalogram (EEG) bisector analysis, tape recordings of sleep were analyzed for two beta, three alpha, three theta, and two delta EEG patterns, as well as for detections of sleep spindles, K-complexes, eye movements, body movements, average electromyogram (EMG), and calculation of seven sleep-waking stages. All three opioids produce a dose-related arousal: they increase EMG and EEG measures of muscle activity, as well as body movements and EEG alpha, while decreasing EEG theta and spindling. These opioids also increase measures of waking state and decrease measures of spindle sleep and REM sleep. Although the 1974 version of the EEG bisector analysis is not exactly comparable to visual analysis, in this design it defined significant drug effects on sleep and EEG. Distinctive bisector analysis patterns are positively correlated with each sleep--waking stage.


Sentence memory and sleep: a pilot study.

In four consecutive weekly sessions 12 subjects, aged 20--25 years, listened to a sentence stimulus before the onset of sleep and were asked to reproduce it after an awakening provoked during stage 2 or REM sleep of the first cycle. Recall of the sentence stimuli heard before sleep was affected by their semantic acceptability, but was unaffected by the sleep type (REM/NREM) or by the length of waking preceding sleep. The differing retention intervals involved for the recall tests after REM and NREM sleep may, however, have masked any effect of the former on recall.


Phasic events of REM sleep: phenomenology of middle ear muscle activity and periorbital integrated potentials in the same normal population.

Both periorbital integrated potentials (PIPs) and middle ear muscle activity (MEMAs) were recorded in a sample of normal male veterans. Activity measures were constructed for each REM period and for the 10 min preceding and following each REM period. PIPs and MEMAs displayed high levels of internight reliability. PIPs were more abundant than MEMAs; in REM the average PIP rate was almost 8 times the average MEMA rate. Within REM, the proportion of MEMAs and PIPs with concurrent eye movement activity was very similar, 66 and 70%, respectively. Both PIPs and MEMAs had higher rates within REM than within NREM. NREM PIP activity was similar both before and after the REM period, whereas NREM MEMA activity was higher before the REM period than after the REM period. Within REM, the distribution of PIPs and MEMAs was similar for REM periods exceeding 30 min. Within-night trends could nor be established for MEMAs due to limitations of the recording technique; PIPs, however, were generally highest in the middle of the night but still higher at the end of the night than at the beginning. Correlations with psychometric test data showed PIP rate in REM to be related to the psychoticism scale of the Minnesota Multiphasic Personality Inventory and MEMA rate in REM to be inversely related to the Barron Ego Strength scale. No associations were found between the psychometric data and measures of PIP and MEMA leakage from REM into NREM. The potential for pathological intrusion of REM phasic events into the waking stage might be conceptualized in terms of a general ungating of activity with PIP and MEMA rates increasing in both NREM and REM alike.


Effect of thyroxine on sleep in the rat.

The relationships between daily sleep time and metabolic rate both in phylogeny and ontogeny have suggested that sleep functions in the regulation of an animal's energy expenditure. Smaller animals may sleep more to conserve more energy. The present study investigated whether, in individual animals, sleep increased in response to the administration of thyroxine, which increased metabolic rate. The sleep, motor activity, food intake, and weights of rats were sampled over a base-line period of 2--4 weeks, during 2--6 weeks of daily administration of thyroxine (150 or 300 microgram/kg), and during a recovery period of 2--4 weeks. In addition to comparisons among base-line, drug, and recovery periods, comparisons were also made with control animals recorded at the same time. Thyroxine produced the increases in food intake and decreases in weight gain which usually accompany increases in metabolic rate. In spite of this evidence of major systemic adjustments in energy input (increased food intake) and energy output (decreased growth) in response to the increased energy expenditure of a raised metabolic rate, there was no change in total sleep time. Therefore, the results do not support the theory that sleep adapts to prevailing energy demands. Although the rate of cycling through the sleep stages is also related to metabolic rate both in phylogeny and ontogeny, in this study thyroxine did not change the period of paradoxical sleep rhythm.


Home and laboratory dreams: four empirical studies and a conceptual reevaluation.

Four empirical studies were presented on how the setting in which dreams are collected may influence the reporting and content of dreams of children on REM awakenings. In these studies it was shown that (1) with constant dream-sampling procedures, home and laboratory did not differ in the degree to which dream reports could be elicited from 4/5 year old children; (2) the rate of truly spontaneous reporting of dreams at home by 6/7 year olds is so minimal as to render these dreams an unfit base for generalization to typical dream life at these ages; (3) with constant dream sampling procedures, home and laboratory reports were not significantly different in content for 10/11 year olds; (4) results typically observed in uncontrolled home versus laboratory studies are replicable at the child level (ages 12/13), but probably reflect biased recall at home rather than effects of REM-monitoring procedures in the laboratory. The extant literature on home versus laboratory dream differences was reviewed, and it was concluded that even for normative dream studies, the REM-monitoring procedure is the method of choice.


The actioculographic monitor of sleep.

The actioculographic monitor (AOGM) is a newly developed, flexible system for sleep recording using three criteria: eye movements, body movements, and submental electromyogram. The signals are recorded on a four-channel Medilog casette tape and displayed by a mingograph at very low speed. The record is then scored after certain criteria for stages wake, nonrapid eye movement, and rapid eye movement sleep. High statistical correlations were obtained when scoring of the AOGM was compared to scoring of standard polygraphic sleep recordings.


Mitogen-induced differentiation of human CLL lymphocytes to antibody-secreting cells.

Peripheral blood lymphocytes from nine patients with chronic lymphatic leukaemia (CLL) have been stimulated by photohaemagglutinin, pokeweed mitogen and Epstein-Barr virus. Our results provide proof that the leukaemic B cells in CLL are capable of responding to mitogenic signals by producing and secreting antibodies, transforming into blast cells, and, probably, increasing their rate of DNA synthesis.


ACTH-adrenal system and exogenous steroids.

The results obtained from a research on 100 patients of both sexes affected with bronchial asthma, chronic obstructive bronchopathy, rheumatoid arthritis, haematological disorders and some other pathological forms, all depending on steroid therapy, are reported in this paper. The treatment was carried out using a new chronopluricorticoid (Dutimelan 8 15). The clinical efficiency is comparable to that obtained by the traditional steroid treatment with the advantage that symptoms of hypercorticism were absent or insignificant. The authors believe that such excellent results are to be attributed to the particular chronopharmacological characteristics of this preparation.


The preservation of medically important fungi.

Details are given of a method, first described in 1962, for preservation of fungi in anhydrous silica gel. It is shown that the procedure is suitable for use with dermatophytes, yeasts and other fungi of medical importance and that they may be preserved for periods of 5 years or more.


Increased mutation rates in doubly viral transformed Chinese hamster cells.

An untransformed parental Chinese hamster cell line, one polyoma transformed clone, and two clones containing both SV40 and polyoma DNA were tested for the appearance of variants resistant to 6-thioguanine or oubain. The frequency of such variants was found to be highest in doubly transformed cells. The mutation rate at these loci was correlated with the level of transformation. The mutants observed were stable and had low reversion frequencies with no gross cytogenetic changes. These results establish a quantitative basis  for the evaluation of phenotypic variability observed in transformed cell populations.


Expression of late viral functions in SV40-infected rat-monkey somatic cell hybrids.

Somatic cell hybrids between a rat hepatoma cell line and an SV40-transformed African green monkey kidney cell line (T22 TK-) have been isolated and their response to infection by SV40 virus characterized. The parental rat cells are nonpermissive to productive infection by SV40, while the parental monkey cells, which are T-antigen positive and capsid antigen negative, are permissive to productive infection. Hybrid clones, which had segregated monkey chromosomes, were infected with SV40 virus, and some clones were found to be negative for SV40 capsid antigen production while other clones were positive. Capsid antigen-positive clones production while other clones were positive. Capsid antigen-positive clones produced little or no form I SV40 DNA, no infectious virus, and were not capable of rescuing SV40 from SV40-transformed mouse cells. Such hybrids, which produce late viral proteins in the absence of mature progeny viral DNA may be useful in the study of control of SV40 RNA transcription.


X chromosome inactivaton and SV40 transformation of mammalian cells.

Five embryonic mouse cultures and one human fibroblast culture were transformed with SV40. The cultures were studied cytologically to see if the normal pattern of sex chromosome replication was maintained in SV40 transformed cells. Characteristic late replication patterns were observed for both the X and Y chromosomes, and there was no evidence for loss of the inactive X chromosome, even in cells with 4 or more X chromosomes. The human line was heterozygous at two X-linked loci and a clonal analysis showed that the expression of X-linked genes was not affected by SV40 transformation.


Studies on cell transformation.

Seven different transformation stigmata of the transformed CHO cell line, including morphological characteristics, growth behavior, cell membrane biochemical properties, and failure of fibronectin deposition, are reversed by addition of cAMP derivatives to the medium. Simultaneously the microtubular pattern changes from a sparse, relatively random set to an orderly arrangement of tubules largely parallel to each other and to the long axis of the resulting fibroblastic cell. Agents like colcemid and cytochalasin B, respectively disorganizing microtubular and particular microfilamentous structures, prevent at least certain aspects of the reverse transformation reaction induced by cAMP in interphase cells. It is proposed that malignant transformation can be effected by damage to the microtubular and microfilamentous structures which changes cell constitution and behavior in two ways: (1) chromosomal instability is introduced which promotes continuous selection for variants better able to resist environmental signals to limit reproduction and (2) a variety of metabolic defects in biochemical processes such as specific membrane functions are introduced which may alter the growth responses of the cell. This picture offers a reasonable explanation for a number of aspects of normal and malignant cell behavior.


Biphasic effects of ethanol on behavioral and electrophysiological parameters can be explained by functional organization of neural networks.

The biphasic effect of ethanol on the CNS is usually explained by assuming that excitatory and depressant effects are mediated by different mechanisms. The present alternative explanation states that ethanol depresses the activity of all neurons to the same extent, but that the biphasic effect can result from the functional organization of the neural network concerned. Current experimental evidence is in accordance with both explanations.


Oscillatory potentials in the electroretinogram.

This paper presents an analog circuit model of the electroretinogram b-wave and subsequent oscillations postulated to result from the depolarization of the retinal glial cells produced by potassium. The system is described by a second-order equation which oscillates under conditions of light adaptation. With dark adaptation the oscillations undergo attenuation. Light falling on the retina ultimately liberates potassium ions and this ionic current is considered to be the system input. A first approximation is an exponentially decaying current which varies depending upon light background, flash intensity and various drug manipulations. This model is a reasonable fit to the electroretinogram over considerable light background levels and is suggested as a useful tool in the study of various pharmacological effects on retinal electrophysiology.


Uveitis associated with sarcoidosis and angiotensin converting enzyme.

Sarcoid uveitis is usually a presumptive diagnosis based on the simultaneous presence of uveitis and clinicoradiographic or histologic findings of sarcoidosis. The elevation of serum ACE in some patients with granulomatous uveitis is strongly presumptive of sarcoid uveitis even in the absence of these findings. Serum ACE may prove to be a useful indicator of ocular sarcoid in the absence of otherwise undetectable systemic disease.


Experimental intraocular malignancy: the effect of intracameral perfusion.

Transplantable Brown-Pearce carcinoma was adapted successfully in the rabbit anterior chamber. Regression of tumor growth was attained on tri-weekly perfusion of the AC with 10 micromolar of methotrexate. Tumor cyclic nucleotide phosphodiesterase (PDE) and protein activator were found to be markedly depressed during the course of chemotherapy and the PDE cAMP/cGMP ratio was similarly altered. Corroborative light and electron-microscopic studies showed specific alterations of intracellular organelles in relation to MTX and tumor cell death. These findings suggest that metabolic pathways of cyclic nucleotides are important biochemical modulators of neoplastic cells. The method of intraocular perfusion precludes systemic toxic effects and avoids compromising the animals' immunocompetence.


[Production and study of the immunogenic properties of a bivalent inactivated vaccine against mucosal disease (bovine viral diarrhea and infectious rhinotracheitis)].

Bivalent inactivated vaccine against mucous disease (MD) and infectious rhinotracheitis (IR) in cattle was produced from cell cultural MD and IR virus suspensions. The vaccine was concentrated on aluminium hydroxide, inactivated by ethanol and is without residual virus. Saponine in final 1:1500 dilution is added as supplementary adjuvant. Immunogeneity of the vaccine was tested on 10-month-old calves, which had shown full resistance against experimental infection with virulent strains of both viruses. Testing on calves for harmlessness by use of a five-fold higher vaccine dose indicated complete tolerance of the vaccine. The prophylactic effect of the vaccine applied in practical work to directly threatened with immediate MD and IR infection cows, including pregnant ones, consisted in reduced number of cases of abortion, of inborn malformations, in lower neonatal calf death-rate, etc. No disturbances were observed following two-fold vaccination of the animals, a fact proving its harmlessness. The positive results of the studied vaccine allow its further application in the combined prophylaxis of MD and IR in calf fattening and breeding complexes.


[Chronic progressive interstitial pneumonia in sheep (adenovirus pneumonia)].

Adenoviral infection was established in imported specialized meat strains of sheep Ile de France and Romni marsh. The disease followed a course typical of chronic interstitial progressive pneumonia with reduced tonus, loss of body weight, and high lethality. The disease was serologically proven by the discovery of specific complement binding antibodies in dynamics against the adenovirus. Titers varied from 1 : 2 to 1 : 32. The infestation was reproduced on sheep and lambs by tracheal infection with ultrafiltrate of changed lung tissue and bronchial lymph nodes. Morphologically the infection manifests itself by considerable hypeplastic processes in the lungs and less pronounced ones in the liver and kidneys. The typical changes indicated are accumulation of lymphoid cells and histocytes in the interstitial tissue, as well as adenomatous expansion of alveolar and bronchial epithelium. Considerable cell polymorphism is observed in the alveoles along with presence of gigantic cells and cell syncytia of desquamous epithelial cells, alveolar macrophages and intranuclear inclusions in some of these cells. Complications of Past. Haemolytica with developing catharalsuppurative pneumonia were observed in some cases.


[Dovenix and Acedist residues in the tissues and organs of guinea pigs and their effects on biochemical blood indices].

It was established that the antihelminth preparations dovenix and acedist are deposited in all inner organs and muscle tissues of laboratory animals (guinea-pigs) treated once with doses 0.1 cm by injection and 100 mg per os. Residual quantities of the named antihelminth preparations were found on the 6th to 57th day post treatment in the liver, lung, heart, kidneys, spleen and muscles. On the 61st day no residual quantities of the preparations were assessed in the muscles and inner organs. The hematological investigations and proteinogramms of experimental animals treated with dovenix and acedist do not reveal significant changes. In guinea-pigs treated with acedist a certain reduction of the beta- and gamma-globulin fractions is observed, while in those treated with dovenix the beta-globulin fractions are reduced.


[Resistance of the Aujeszky's disease virus to gamma ray irradiation].

Investigations on the resistance of Aujeszky's disease virus to gamma irradiation from a 60Co source were performed. The resistance of the virus was tested in vitro in a fluid medium of 50% Henk's balanced saline solution, 50% Eagle's medium MEM and a supplement of 10% normal calf serum as well as its resistance on artificially virus infected pig skin and greasy wool of sheep. It is established that virus inactivation in the fluid medium occurs following 1.6 Mrad gamma irradiation. The irradiated with lower than 1.6 Mrad gamma doses live virus particles form smaller than and with irregular oval shape and dentated plates as compared to the non-irradiated control virus. 1.5 Mrad gamma rays are needed to inactivate the virus on pig skin and greasy wheep's wool. The presence of lanoline in greasy sheep's wool, due to the alkaline pH (9--13) it produces, has an effect on the vitality of the virus both in the control and in the experimental samples.


Studies on the substrate specificity of the DNA methylase activity from Escherichia coli K-12.

A partially purified extract of DNA methylases from E. coli K-12 containing DNA-adenine as well as DNA-cytosine methylase activities has been examined with respect to different DNA species as substrates. The results show that the natural content of 6-MAP) in the applied DNA represses the DNA-adenine methylase activity. On the other hand, 5-MC, already present in the substrate does not influence the activity of the DNA-cytosine methylase. DNA from Micrococcus radiodurans, which is completely free of methylated bases served as comparison. Since netropsin preferentially binds to AT-rich regions of DNA, the influence of this oligopeptide antibiotic on the methylation of DNA was investigated. As expected the antibiotic predominantly inhibits adenine methylation of DNA. The degree of inhibition depends on the molar ratio of netropsin to DNA phosphate.


Effects of drugs that influence eucaryotic motile processes on motility of Cystobacter fuscus (Myxobacterales).

Cytochalasin B, dbcAMP and EGTA were found to inhibit the motility of Cystobacter fuscus completely whereas colchicine and cAMP only reduced velocity of movement.


[Use of gen5 and gen6 ts-mutants of phage T3 as indicators of inhibitors of DNA synthesis].

In an enzyme-specific drug screening system nalidixic acid and 3'-FTdR, inhibitors of DNA synthesis, both reduce the growth of wild type and temperature-sensitive point mutants of phage T3 with different efficiencies. The wild type shows the strongest sensitivity against the drugs, while an exonuclease mutant is the most insensitive variant. The DNA polymerase mutants exhibit an intermediate degree of inhibition. The anthracycline antibiotics violamycin BI and adriblastin which preferentially inhibit RNA synthesis show the same degree of inhibition for all mutants. This is true also for the RNA synthesis inhibitor lambdamycin, which is identical with chartreusin. The protein synthesis inhibitors chloramphenicol and o-phenanthroline, a chelating agent, impair all mutants to the same extent. Our data confirm the hypothesis that structural variants of essential viral enzymes, when compared with the wild type should reveal different sensitivities against specific inhibitors and show that this T3 system could be used for the indication of specific inhibitors of DNA synthesis.


Corticotroph cells of the human pituitary in old age.

This study was undertaken, utilizing immunocytologic methods and in particular the immunoperoxidase technique, to evaluate corticotroph cell morphology and distribution in the pituitary glands of aged subjects dying of various diseases. Early histologic studies did not allow for reliable identification of the different adenohypophysial cell types. Corticotroph cell distribution within the gland is described as well as its morphological characteristics. Our findings substantiate conclusions based on biochemical investigations that corticotroph cells remain unchanged and function normally in aged subjects.


Molecular cloning of the restriction fragments derived from double EcoRI/PstI digestion of the calf satellite I DNA and their restriction analysis.

Calf satellite I DNA digested with EcoRI and PstI gives three fragments 643, 621 and 83 bp long. Two of them, the 643 and 621 bp were cloned using pBR 322 vector and analyzed by means of the MspI and HaeIII restriction enzymes.


Expression of aspartokinase, dihydrodipicolinic acid synthase and homoserine dehydrogenase during growth of carrot cell suspension cultures on lysine- and threonine-supplemented media.

Reduction in the amounts of activity of the first enzyme, aspartokinase (EC 2.7.2.4) and two branch-point enzymes, dihydrodipicolinic acid synthase (EC 4.2.1.52) and homoserine dehydrogenase (EC 1.1.1.3), located in the pathway for the synthesis of aspartate-family amino acids, occurred when cell suspension cultures of Daucus carota L. var. Danvers were grown in media containing 2 mM threonine or 2 mM lysine, endproducts of the pathway. Activity of the lysine-sensitive form of aspartokinase was decreased when cells were grown in medium containing lysine and the activity of the threonine-sensitive form was decreased when cells were grown in medium containing threonine. Activity of the branch-point enzyme leading to threonine synthesis, homoserine dehydrogenase, was decreased up to 70% in specific activity (units/mg protein) and relative activity (units/g fresh weight) when cells were grown in media containing lysine or threonine. Threonine had no effect on the relative activity of dihydrodipicolinic acid synthase, but decreased its specific activity. Lysine decreased the relative activity of the synthase by up to 40%, but had little effect on its specific activity. The decreased activities of the enzymes were apparently not due to binding of the inhibitory amino acids to the enzymes since homogenization of cells in buffer with 2 mM lysine and threonine did not decrease the measurable enzyme activities. These and other results presented suggest that both forms of the aspartokinase activity and homoserine dehydrogenase activity can be altered by supplementing the growth medium with lysine or threonine.


Permeability of plasma membrane vesicle to ouabain and Mg2+ as a factor determining rate of binding of ouabain to Na+ and K+ dependent ATPase.

Na+, K+-ATPase of the plasma membrane isolated from sheep kidney medulla exhibits functional asymmetry for the cardiac glycoside ouabain. In this vesicular membrane preparation the rate of binding of ouabain was slow (time constant greater than 60 min) when the vesicles were incubated in the presence of isotonic sucrose. Upon treatment of the preparation with hypoosmotic shock or phospholipase A the initial rate of ouabain binding was enhanced at least 3 fold. In equilibrium a concentration of the ouabain-enzyme-complex was obtained which was about twofold that of the untreated vesicles. This result suggests two types of ouabain binding sites with an approximate stoichiometry of 1 to 1. The stoichiometry seems to be maintained at high concentrations of ouabain where binding curves show a biphasic time course. Additional information about heterogeneity of binding sites comes through experiments in which the vesicles were treated with Mg2+ prior to the addition of ouabain. A minor fraction of the binding sites were occupied by ouabain only after longtime incubation with Mg2+.


II. On the construction of a phylogenetic tree.

For the construction of a phylogenetic tree in algorithm is described. This, allows the correction of the original data and the proper selection, in each step of the process, of the nearest neighbours of a common ancestor.


On the action of diethyldithiocarbamate as inhibitor of copper-zinc superoxide dismutase.

The rate constants of the reactions between pulse radiolytically produced superoxide radicals and the Cu(II) chelate of diethyldithiocarbamate were determined at pH 7.0. It was found that diethyldithiocarbamate forms a copper complex, which as no dismutating activity. The removal of the protein bound copper in superoxide dismutase by diethyldithiocarbamate yields the same effect as coordination of the copper in the enzyme.


[Characterization of mitochondria of Aspergillus niger from mycelium growing in surface and submerged culture conditions (author's transl)].

This paper deals with studies on tightly coupled mitochondria present in the active citric acid producing mycelium of Aspergillus niger growing in surface and submerged culture. A special homogenizer had been used for rapid extraction of mitochondria. Observation in the electron microscope indicated that some of the isolated mitochondria were probably damaged during preparation. Nevertheless, the crude mitochondrial fraction was capable of coupling phosphorylation to the oxidation of three different substrates tested viz., succinate, citrate and NADH. It was found that yield of mitochondria was greater in submerged mycelium than in the surface mycelium of A. niger.


[The resistance of a maternally-derived immunity of baby mice against mousepox--experiments regarding a special possibility of immunization with orthopox viruses (author's transl)].

In the scope of research concerning new proplylactic possibilities against mousepox we tested the resistance of a passive immunity of baby mice whose mothers had been immunized before or during pregnancy several times with either the vaccinia virus strains MVA, CVB or the so-called attenuated ectromelia virus strain "Hampstead" respectively. The vaccines CVB and "Hampstead" were found to cause the production of haemagglutination-inhibiting antibodies only. A passively-transmitted immunity of baby mice was proven to be effective against an intraperitoneal challenge-infection with a virulent ectromelia virus strain for as long a period as 3 weeks after birth, if their mothers had been vaccinated with either of the following: 10(2,0) and 10(4,0) KID50 of the "Hampstead" strain, or one of the vaccinia virus strains in a dosage 100-times as large. The vaccination did not lead to an intrauterine infection of the fetuses. After 224 tissue culture passages in mouse fibroblast cells (line 3T6), THE "Hampstead" strain was found te be still virulent for baby mice; however, female adult mice which had been intraperitoneally inoculated with 10(2,0) and then 10(4,0) KID50 two weeks later demonstrated no signs of mousepox. The possibility for using this ectromelia virus strain instead of vaccinia virus for active immunization against mousepox of young animals under the protection of a homologous passive immunity is discussed.


[Sleep stages and growth hormone in manganese poisoning. Effects of L dopa].

Sleep characteristics are studied in 7 patients with sequelae of manganese intoxication and in 8 controls. A reduction in the length of the REM stage was observed, which could be prolonged by L Dopa administration, but without achieving values comparable to the controls. No changes were noted in the non-REM stage of sleep. These results are similar to the ones obtained in Parkinson's disease, suggesting clinical and probably biochemical similarities between the 2 conditions. The blood concentrations of growth hormone during sleep in the manganic patients was lower than in the control group, and no significant variations were noted after L Dopa administration.


Cerebral embolism in nonbacterial thrombotic endocarditis associated with carcinoma. A clinico-pathological study.

The clinical and pathological features of 24 patients with cerebral emboli complicating 66% of our cases of nonbacterial thrombotic endocarditis (NBTE) associated with carcinoma are reviewed. Twelve patients were admitted for a cerebrovascular accident (CVA) while 4 patients developed a CVA during hospitalization. Transient ischemic attacks preceded the CVA in 3 patients. More often the CVA took the form of a single sudden accident. Cerebral infarcts however were generally multiple and hemorrhagic and varied in size and age. In 4 patients large softenings were directly responsible for death. 8.6% of cerebral embolisms were caused by NBTE and in 10 patients cerebral embolization was the first symptom of a carcinoma. The frequency of NBTE in ovarian carcinoma even in the absence of metastases may motivate a more aggressive approach towards unexplained cerebral embolism.


Chorioretinitis of the newborn with herpes simplex virus type 1. Report of a case.

A disseminated herpes simplex virus (HSV) infection involving the central nervous system and accompanied by chrioretinitis in a a 3-week-old girl is described. The aetiologic diagnosis was established on the basis of vrius isolated from skin vesicles, and a significant rise in complement fixing antibodies to HSV type 1. The mode of transmission of the virus to the infant apparently was direct contact with an oral lesion in the mother, that was present at the time of delivery. The patient survived but became blind and microcephalic, with severe neurological sequelae. The virus isolated was identified as HSV type 1, which is an infrequent finding in herpetic chorioretinitis of the newborn.


In vitro production of steroid cataract in bovine lens. Part II: measurement of sodium-potassium adenosine triphosphatase activity.

Fresh calf lesnses incubated in nutritive media containing dexamethasone phosphate or ouabain in concentrations ranging from 1 X 10(-4) M to 1 X 10(-8) M developed cortical opacification and showed significant inhibition of Na(+)-K(+) ATP'ase activity. Over a 3-day incubation period the decrease in Na+-K+ ATP'ase activity correlated well with the observed decrease in light transmission. The degree of enzyme inhibition and decrease in light transmission varied directly with the concentration of dexamethasone phosphate and ouabain, with significant changes observed at 'physiologic' and 'pharmacologic' concentrations of these agents. Lenses incubated for 4 days in dexamethasone phosphate or ouabain showed substantial increases in water content as well as an increase in Na+ and a decrease in K+ concentration. These data suggest that inhibition of the cation pump may play a significant role in the formation of steroid cataract in vitro.


Effect of starvation on tissue and serum gluconeogenic enzymes, alkaline phosphatase and tissue glycogen in the freshwater catfish, Heteropneustes fossilis (Bloch).

The influence of starvation has been studied on tissue and serum G-6Pase F-D-Pase and alkaline phosphatase activities and on the muscle and liver glycogen content of the freshwater catfish H. fossilis (Bloch). A marked increase in G-6Pase and F-D-Pase activities and a fall in the muscle and liver glycogen content recorded during 40 day starvation. The rise in gluconeogenic enzymes during starvation may be due to glucocorticoid stimulation. Alkaline phosphatase activity was found to decline markedly during starvation. The decline in enzyme activity is attributed to some factors like a fall in the rate of synthesis caused by lowered metabolic demands and to electrolyte imbalance caused by tissue overhydration. The fall in glycogen content may be related to the starved condition of the fish. Elevation in glycogen content and alkaline phosphatase activity and a fall in gluconeogenic enzymes were noted when feeding had been resumed.


Micro-arousals during nocturnal sleep.

In 8 young adult human subjects EEG- and polygraphic characteristics of transient shifts towards arousal (micro-arousal, MA) have been studied during sleep under five different experimental conditions in 40 night sessions. Out of the five applied experimental situations, two (psychostimulant application and sensory stimulation) resulted in a shift of the balance between the systems of sleep and arousal towards an increased activity of the arousal system, while an other condition (rebound following partial sleep deprivation) led to an opposite change to a rise in "sleep pressure". An inverse correlation has been found between the frequency of MA and the depth of sleep, a finding consistently observed in every subject and in every experimental situation. During the process of sleep periodic changes in the dispersity of MA could be seen; the number of MA-s decreased and increased according to the descending and ascending slope of the sleep cycles. During the ascending slope of cycles there was a coupling between the occurence of MA-s and the changes of phases. Increases in the level of activation and in sleep pressure did not influence the occurrence of MA-s. Increasing the tone of the arousal system in chemical way, or by means of enhancing the phasic sensory input resulted in a reduction of the difference between the number of MA on the descending and ascending slopes of cycles. During the phases of sleep, the spontaneous occurrence of MA-s went parallel with the possibility to evoke MA-s by sensory stimuli. These data show that MA is a regular phenomenon of nocturnal sleep; MA manifests itself as a result of phasic functioning of the reticular arousal system and plays a role in the organization of those periods of the sleep cycle, which tend toward arousal. It is suggested that MA-phenomenon is considered a standard measure of sleep and that it could represent an indicator of the function of the arousal system controlled by external or internal mechanisms during sleep.


Effect on renal function and renin secretion of noradrenaline infused into the renal artery.

Effect of noradrenaline on renal function and renin secretion was studied during infusion into the renal artery of anaesthetized dogs. Experiments were performed with or without alpha or beta receptor blockade. Noradrenaline infusion resulted in a significant elevation of renin secretion associated with marked vasoconstriction. Urine flow rate, the filtered and excreted amounts of sodium were diminished due to the decreased GFR. Alpha receptor blockade suppressed renin secretion in the presence of changes in renal haemodynamics. The simultaneous infusion of noradrenaline enhanced renin release without affecting renal haemodynamics or reducing Na-excretion. Following simultaneous inhibition of alpha and beta receptors renin secretion dropped markedly; there were no further changes in either renin secretion or renal haemodynamics upon the simultaneous administration of noradrenaline. Based on the present findings it is suggested that renin secretion is controlled by both alpha and beta receptors. Beta receptor simulation exerts a direct action, whereas alpha stimulation appears to be mediated in part by indirect mechanisms such as renal haemodynamics.


Effect of KCN on the motility of Treponema pallidium.

Virulent Treponema pallidum has been shown to survive in KCN-containing artificial medium. Oxygen uptake being sensitive to cyanide, the observation indicates that treponemes do not have a cytochrome oxidase system. KCN had a killing effect only at a 15--30 mM final concentration. The data show that the Budapest strain of T. pallidum is an anaerobic organism.


Functional heterogeneity and T cell-dependent activation of macrophages from murine sarcoma virus (MSV)-induced tumors.

In studies on the functional activity of macrophages isolated from murine sarcoma virus (MSV)-induced tumors, we have found that these cells may suppress immune responses as well as act as effector cells against the tumor. Previously, we reported that macrophages from the tumor could inhibit the antitumor response by suppressing proliferation-dependent immune functions. Here, we demonstrate that macrophages can also suppress the production of migration inhibition factor (MIF) and macrophage activation factor (MAF), two lymphocyte activities that are independent of cell proliferation. Conversely, we and others have found that macrophages from the tumor can exert an antitumor, cytolytic effect. In this study, using 1 g velocity sedimentation separation techniques, we have been able to identify 2-3 subpopulations of cytolytic macrophages in regressing tumors but in progressing tumors, only the smallest subpopulation of macrophages was active. T cells appeared to be required for activation of macrophages within the tumor, since MSV tumors induced in athymic, nude mice did not contain cytolytic macrophages.


Mechanism of resistance of mice to syngeneic methylcholanthrene induced fibrosarcomas.

Mice become resistant to challenge with certain fibrosarcomas when bearing a tumor graft (concomitant immunity) or after injection of a low, non-tumorigenic dose of cells (sinecomitant immunity). Resistance to footpad challenge was depressed or abolished by treatment with carrageenan, niridazole or reserpine, or by sublethal irradiation, all of which also depressed delayed-type hypersensitivity (DTH) reactions. Immune lymphocytes initiating tumor-suppressive reactions in the feet of non-immune mice were Thy-1+, Ly-1+, Ly-2- and Ly-3-. Injection of tumor cells into the peritoneal cavities of immune mice specifically elicited an influx of macrophages. There was evidence of macrophage stimulation in tumor-immune mice. In vitro, anti-tumor effector cells lacking individual tumor specificity could be detected among the resident peritoneal cells of tumor-immune mice and among peritoneal exudate cells of non-immune mice. The expression of acquired resistance to some tumors may involve reactions akin to DTH in which a specific reaction triggers an accumulation of nonspecific effectors.


Human placental adenosine kinase: purification and characterization.

Human placental adenosine kinase has thus been purified 3600-fold and characterized with respect to molecular weight, substrate specificity, divalent cation requirements, pH optimum, isoelectric pH, and kinetic properties. These data contribute to the information currently available about the regulation of adenosine metabolism, information critical for an understanding of the biological properties of adenosine.


The use of amniotic membrane in acute massive full-thickness loss of the abdominal wall from clostridial myonecrosis.

A patient who developed clostridial infection of the abdominal wall following laparotomy for small bowel perforation is described. Immediate debridement resulted in the loss of the entire abdominal wall. Dressing of the exposed viscera during the period of stabilization and prior to reconstruction was achieved using amniotic membranes.


[Study of the effectiveness of morantel tartrate (Exhelm II) in bovine gastrointestinal strongylosis in Ivory Coast].

A study of the seasonal incidence of the strongyles of the bovine digestive tract, under natural climatic conditions (rainfall, humidity and temperature) and the use of Morantel Tartrate (Exhelm II, Pfizer, France) for its control was conducted, in the North of the Ivory Coast during 1976-1977. Young calves belonging to the local breeds Baoulé, N'Dama, and cross bred Zebus-taurins employed for this study were located in the East, West, North and Centre of the Korhogo area. On the basis of the results obtained it could be concluded that: the infestation rate calculated on the basis of egg output, increases during the rainy season, between April and October and decreases during the dry season namely between November and March/April; the animals treated twice or three times with Exhelm II expelled considerably less E.P.G., in comparison to that of the control group. It is suggested that this drug may be used at the dosage rate of 7,5 mg per kg live body weight under field condition at least twice in a year, at the beginning and at the end of the rainy season. A third treatment using this same anthelmintic during the rainy season does not seem to be more profitable than two treatments. As such this third treatment should not however be considered as being essential. N'Dama and Baoule of that region have mixt infection of strongyles comprising of Trichchostrongylus 38%, Oesophastomum 30%, Cooperia 17%, Bunostomum 16%, Haemonchus 15%, Hematodirus 2% and Ostertagia 0,2%.


Response of susceptible versus immune chicks to killed, live-modified, and wild infectious bursal disease virus vaccines.

Modified infectious bursal disease virus (IBDV) administered ocularly to either susceptible or passively immune chicks did not induce protection against bursal atrophy by wild IBDV, while intramuscular (IM) or intrabursal (IB) injection protected susceptible chickens. No protection was obtained in passively immune chickens vaccinated IM or IB. Susceptible and passively immune chickens vaccinated with a single dose of killed-virus suspension (KVS) did not become resistant to wild IBDV challenge. A killed-virus multiple emulsion (KVME) induced partial protection (3/5) against challenge 28 days after subcutaneous vaccination of three-day-old susceptible chickens. At 35 and 42 days old, all susceptible chickens were protected by KVME. Protection by passive antibodies was observed in unvaccinated, KVS-, and KVME-vaccinated chickens challenged at 10 and 17 days of age. Protection was only partial in passively immune chickens vaccinated with KVME after passive antibodies were no longer protective to unvaccinated and KVS-vaccinated chicks. Modified IBDV was detected two days post-IB inoculation only in the bursa of Fabricius of susceptible chickens, while wild IBDV was found also in the thymus, spleen, and blood. No evidence of virus was found in immune chickens inoculated IB with modified IBDV. The growth of wild IBDV was limited to the bursa of Fabricius in immune chickens. Furthermore, the intensity of fluorescence, as well as the number of positive bursa cells, was low when compared with fluorescence in the bursa of susceptible chickens infected with wild IBDV.


The prevention of experimentally induced necrotic enteritis in chickens by avoparcin.

Four groups of about seventy 2-week-old broiler chickens were challenged with a Clostridium perfringens Type A isolate. Inclusion of avoparcin at 20 ppm in feed prevented necrotic enteritis in one group, but 10 ppm was only marginally effective. Bacitracin at 110 ppm also prevented the disease. Necrotic enteritis was successfully reproduced in untreated control birds.


Marek's disease tumor-associated surface antigen (MATSA) in resistant versus susceptible chickens.

The appearance of MATSA-bearing cells in the spleens of genetically susceptible (P-line) and resistant (N-line) chickens followed similar patterns at 4 and 6 days postinoculation (PI), with 4 to 10% of the cells positive in indirect fluorescent-antibody tests. Levels of MATSA-positive cells at 10, 14, and 21 days PI continued at 6-8% in P-line birds but dropped from about 8% to below 1 or 2% in N-line birds. This pattern was seen also in virus-isolation rates from the same spleen samples. Viral internal antigens (VIA) were seen equally and with decreasing incidence in both groups at 4-10 days PI. VIA were not detected at 14 days but reappeared at 21 days, with higher levels in P-lines than inN-lines. It was concluded that genetic resistance to Marek's disease is not related to events which result in production of the putative tumor antigen, MATSA, but rather to host-controlled factors which terminate those events.


Loss of cell-associated Marek's disease vaccine titer during thawing, reconstitution, and use.

Tests of the effects of mismanagement practices observed in hatcheries showed that very subtle deviations from the explicit directions accompanying Marek's vaccine can cause dramatic losses of vaccine potency. Such mismanagement was found to result in 14 to 97% loss of vaccine titer. It appears that procedures used in hatcheries for the handling of cell-associated Marek's vaccine should be closely monitored on a continuous basis to prevent a gradual drift from correct handling.


Characterization of avian reoviruses isolated from the synovia and breast blister.

Reoviruses Texas, S 1133, UMI 203, and WVU 2937 induced swelling of the foot pad and inflammatory changes in the synovial membranes when inoculated in the foot pad of 2-week-old gnotobiotic chicks. From differences in virus neutralization as measured with geometric mean (R) value, all four viruses are subtypes of a single serotype. The cell-associated and cell-released virus growth curves were similar, with a lag phase of about 15 hours and a log phase of 15 to 21 hours. Viral RNA synthesis reached a peak in 5 hours and was active at 14 hours but not at 18 hours. In 90 minutes at 60 C the titer of each virus had decreased about 4 logs.


Thiourea derivatives as specific inhibitors of picorna viruses.

30 compounds with antipicorna virus activity were selected from 173 N,N'-disubstituted thiourea derivatives. The spectrum of antiviral activity was determined in vitro using entero, FMD, rhino and EMC viruses. Structure-activity relationships were studied. Several compounds produced marked activity against coxsackie viruses A and B infection of mice and FMD infection in mice and guinea pigs. N-Phenyl-N'-3-hydroxyphenyl-thiourea (PTU-23) inhibited poliovirus production by more than 99% without influencing the FL host cell. EMC virus was readily inhibited by PTU-23 in Kreb-II cells. Virus RNA synthesis was significantly reduced. Under the effect of PTU-23 the amount of 37S ssRNA extracted from EMC virus infected cells was considerably more reduced than that of the 20S ds RNA. PTU-23 did not influence the activity of virus-induced RNA polymerase in a cell-free system.


Duck plague: a carrier state in waterfowl.

Healthy waterfowl were found to be carriers of duck plague (DP) virus. Black ducks (Anas rubripes) and Canada geese (Branta canadensis) surviving a natural outbreak of DP at Coloma, Wisconsin, in 1973 yielded DP virus in cloacal swabs taken four years postinfection. Experimental infection of previously unexposed mallard ducks (Anas platyrhynochos) with the Coloma strain of DP virus CO-WI (73) also produced cloacal virus shedding for up to four years after infection. A second DP virus strain, LA-SD (73) from the Lake Andes, South Dakota, epornitic, was detected from cloacal swabs of pintail ducks (Anas acuta), gadwall ducks (Anas strepera), wood ducks (Aix sponsa), and Canada geese infected experimentally one year before. The frequency of swabs positive for DP virus varied between individuals within each of the tested species. The amount of detectable DP virus shed was about 100 plaqueforming units of virus percloacal swab. Oral erosions were present in all species tested except Canada geese and gadwall ducks. Erosions occurred at the openings of the sublingual salivary gland ducts. DP virus was isolated from erosions. All ducks with lesions proved to shed DP virus, although not necessarily at the time they had the lesion.


Characterization of Immunosuppression in chickens by infectious bursal disease virus.

Chickens were infected with infectious bursal disease (IBD) virus in ovo or at different times posthatching to 6 weeks of age. The B- and T-cell responses in the lymphoid tissues and blood were examined sequentially to 8 weeks of age by using indirect immunofluorescence. The proportion of B-cells was consistently lower in infected birds than in controls, especially in chicks infected as embryos or at 1 day old. The proportion of T-cells increased following these early infections but was slightly lower in spleen and blood of birds infected at 1, 4, and-6 weeks of age. Serum IgM levels dropped significantly after infection, regardless of the time of infection. IgG levels decreased following early infection but increased after infection at 1 week old or more. The results strongly suggest that B-cells are the target for IBD virus infection.


1H NMR study on the interaction of cupric ion with ribonuclease A.

The reassignment of the 1H NMR C-2 histidine signals of the bovine pancreatic ribonuclease A has required a revision of the 1H NMR data on the role of the different histidines in their interaction with the Cu2+. The results of our measurements carried out at p2H 5.5 and 7.0 reduce the importance of His-12 as main site of interaction. At p2H 5.5 a very strong binding site involves His-119, while a weaker one contains certainly His-105. On the contrary, at p2H 7.0 the histidines 105 and 119 seem to possess binding constants of the same order of magnitude and in addition they provide stronger ligands for the Cu2+ than His-12. The comparison with X-ray data in the crystal shows numerous analogies. Finally, preliminary results on the competitive inhibition effect between the Cu2+ and 2',3'-cytidine monophosphoric acid are discussed.


Interaction of infectious bursal disease and coccidiosis in layer replacement chickens.

During a floor-pen study on anticoccidial programs for layer-replacement pullets, there was a natural outbreak of infectious bursal disease (IBD). The interaction of coccidiosis and IBD was characterized by: excessive deaths from coccidiosis; and apparent failure of some anticoccidial drug treatments. Immunity to coccidiosis was not blocked by the IBD infection, although immunity was less in some treatments than would be expected with the degree of coccidiosis exposure.


Isolation of the lymphocytosis promoting factor-haemagglutinin of Bordetella pertussis by affinity chromatography.

The lymphocytosis promoting factor-haemagglutinin of Bordetella pertussis was isolated from solutions obtained after cell disintegration by a novel affinity chromatographic method using an adsorbent composed of human haptoglobin covalently attached to a Sepharose 4B matrix. The haemagglutinin was bound to the adsorbent at pH 6.5 and eluted by a stepwise change to a pH 10 buffer. A 300--600-fold purification of the haemagglutinin was achieved by this one-step process. The chemical and biological properties of the haemagglutinin isolated by affinity chromatography were found to be similar to those of the protein isolated by other workers from culture supernatants. The affinity chromatographic method was found to be specific for the purification of the lymphocytosis promoting factor-haemagglutinin and no purification of the fimbrial-haemagglutinin of Bordetella pertussis was achieved by the method.


Cobalt-substituted hemoglobin Zürich (alpha 2 beta 263His leads Arg). Oxygen equilibria and EPR spectra.

Cobalt hemoglobin Zürich (alpha 2 beta 263His leads to Arg) has been successfully reconstituted from the apohemoglobin Zürich and cobaltous protoporphyrin IX. The oxygen affinity of cobalt hemoglobin Zurich, as well as that of iron hemoglobin Zürich, were measured in the absence and presence of organic phosphate and Cl-. The overall oxygen affinity of cobalt hemoglobin Zürich was found to be higher and the cooperativity as measured by the n value was smaller than those of cobalt hemoglobin A. Organic phosphate and Cl- affect the oxygen equilibrium properties of cobalt hemoglobin Zürich in a manner similar to that of cobalt hemoglobin A, but to a lesser extant than cobalt hemoglobin A. The EPR spectrum of oxy cobalt hemoglobin Zürich is less sensitive to the replacement of the buffer system from H2O to 2H2O, indicating that the hydrogen bond between the distal amino acid residue and the bound oxygen is not formed in the abnormal beta subunits. The deoxy EPR spectrum of cobalt hemoglobin Zürich is similar to that of deoxy cobalt hemoglobin A, suggesting that the deoxy cobalt hemoglobin Zürich is predominantly in the deoxy quaternary structure (T state).


[Kinetic parameters of Na,K-ATPase in the gills of crabs adapted to freshened sea water].

The dependence of Na,K-ATPase activity on concentrations of ATP, Na+, K+, Mg2+ and ouabain in the membrane preparations of crab gills was studied. The first group of crabs was adapted to freshened (25%) and the second one--to normal (100%) sea water. A 40-day adaptation of crabs to the freshened sea water results in an increase of maximal activity of Na,K-ATPase, but does not affect the enzyme affinity for ATP, Na+, K+, Mg2+ and ouabain, as well as its cooperative properties. It is assumed that adaptation of crabs to freshened sea water is accompanied by an accumulation of Na, K-ATPase in the epithelial cell membranes or crab gills without causing any qualitative changes of the enzyme.


[Reconstitution of electrogenic function of pyrophosphatase isolated from Rhodospirillum rubrum membranes].

The membrane vesicles (proteoliposomes) have been reconstituted from phospholipids and inorganic pyrophosphatase (EC 3.6.1.1) isolated from Rhodospirillum rubrum chromatophores. An addition of inorganic pyrophosphate (PPi) causes a Mg2+-dependent formation of a transmembrane electric potential difference and an uptake of penetrating tetraphenylborate anions by the proteoliposomes. Thus, isolated pyrophosphatase, being incorporated into the phospholipid membrane, functions as a MgPPi-dependent protein generator of the electric current.


Sleep learning during stage 2 and REM sleep.

Pairs of subjects were presented with a 20-item picture series at bedtime. In the latter part of the night, a tape-recorded series of 10 words, the verbal equivalent of half the original series of pictures, was repeated 10 times during either Stage 2 or REM sleep. Morning recall and recognition for repeated words was found to be facilitated following repetition during Stage 2 sleep, but relatively unaffected following repetition during REM sleep. However, adjusting for recall, the number of additional words elicited through recognition was found to be significantly greater for REM repeated words than for Stage 2 repeated words. It was suggested that retrieval limitations, perhaps as a result of REM state dependency, rather than storage inhibition may be the main locus of the initial recall failure. By comparison, Stage 2 sleep would seem to present both a lower barrier to memory storage and retrieval compatibility with wakefulness.


Receptors for angiotensin: a critical analysis.

Angiotensin exerts numerous contractile and secretory effects by activating specific receptors. Recent pharmacological findings obtained with this peptide in various laboratories are analyzed, using the order of potency of agonists and the affinity of competitive antagonists as criteria for the classification of receptors for angiotensin in several systems. The analysis is restricted to experiments in which biological effects have been measured. Desensitization (the third criterion for classification of receptors) is discussed and a new protocol is proposed for its utilization. The analysis reveals that receptors for angiotensin in intestinal and vascular smooth muscles, in the heart, and in the vas deferens are all of the same type, while the receptors mediating the release of catecholamines from the adrenal medulla and those subserving the steroidogenic action on the adrenal cortex remain still unidentified. The recently proposed role of ATI as mediator of renin in the adrenal medulla is not substantiated by pharmacological findings with decapeptide antagonists. Moreover, the utilization of ATI as an agonist to determine the order of potency of angiotensins and the use of SQ 20881 as an inhibitor of the converting enzyme have shown serious limitations and should be reconsidered. The hypothetical role of ATIII as mediator of the renin-angiotensin system in the adrenal cortex, at least in other species than the rat, appears to be supported by the high affinity of heptapeptide antagonists for the adrenocortical receptor. However, these antagonists have generally been compared with [Sar1,Ala8]-ATII, A compound which is definitely inadequate for evaluating the affinities of octapeptides in the adrenal cortex. Therefore most of the data supporting the role of ATIII in this system have to be carefully reconsidered. Analogues of ATII are proposed for using as agonists and as antagonists instead of the natural angiotensins (for determining the order of potency of agonists) and instead of [Sar1,Ala8]-ATII (for measuring the affinities of competitive antagonists).


Permeation of sodium through calcium channels of an insect muscle membrane.

The contribution of Na ions to the electrically excited response was studied in the muscle fibres of mealworm larvae, Tenebrio molitor, using microelectrode techniques. When Ca ions were omitted from the external solution, no action potential could be elicited. However, addition of Na ions to Ca-free medium rendered the fibre excitable again. The amplitude of these action potentials increased with a slope of about 40 mV for a 10-fold elevation of external Na concentrations. Tetrodotoxin had no effect on the initiation of the spike, and Co ions completely suppressed it. Therefore, it seems likely that a Ca-channel, which is utilized by both Na and Ca ions, is the sole factor responsible for the action potential in the mealworm larval muscle fibre membrane.


Purification of soluble hepatocellular carcinoma extracts.

When soluble hepatocellular carcinoma (HCC) extract prepared by 3 M KCl solution was passed through a column of Sephadex G-200, four fractions (designated as I, II, III, IV) could be obtained from the elution profile. By leukocyte migration inhibition assay, the active component of soluble HCC extract which represented tumor-associated antigen was shown to be contained in fractions I and II.


Sulfatide synthesis by neural cell lines.

Several neural cell lines were examined for their ability to synthesize sulfatide and 2',3'-cyclic nucleotide phosphohydrolase, biochemical components characteristic of myelin. The mouse glioma G26 and the rat schwannoma TRM6B actively produced sulfatide, while the rat glioma C6 was inactive, supporting the probable oligodendroglial origin of the G26. In contrast, the C6 cell line had a high level of 2'-3'-cyclic nucleotide phosphohydrolase activity, while TRM6B showed 30% and the G26 75% lower activities. Thus, these two activities appear to be independently regulated.


Effect of prostaglandins on milk ejection.

Prostaglandins (PGs) of type F2 alpha, E1, and E2 have been reported both, to inhibit or to facilitate posterior pituitary oxytocin release in lactating animals and women, and to suppress or to stimulate the mammary myoepithelium. Prostaglandin-induced milk ejection in women and cows has been attributed to central oxytocin release, but no oxytocin blood levels were determined. Moreover, for lactating cows, sows, rabbits, guinea pigs, and rats a direct PG effect on the mammary myoepithelium resulting in milk ejection has been suggested. On the other hand, PGs were found to antagonize the milk-ejection response to oxytocin in rabbits and rats. The mechanisms involved in PG synergism or antagonism of oxytocin-induced milk ejection are not understood. Studies in lactating rats showed that blood pressure active PG doses of F2 alpha, E1, and E2 largely inhibited the intramammary pressure response to oxytocin. Whereas the oxytocin-antagonistic action of PGF2 alpha was not affected by adrenergic blockers (phenoxybenzamine, propranolol), the anti-oxytocin effects of PGE1 and E2 were eliminated after alpha-receptor blockade while the activity of oxytocin increased. Under beta-receptor or alpha- plus beta-receptor blockade, the oxytocin-inhibitory effects of PGE1 and E2 were almost abolished. Mechanisms of PG-induced inhibition of the oxytocin response may involve mammary vascular changes and/or alterations in myoepithelial activity of cyclic adenosine-3,5-monophosphate (c-AMP), cyclic guanosine-3,5-monophosphate (c-GMP), and phosphodiesterase (PDE). It seems unlikely that PGs bring about significant posterior pituitary oxytocin release in rats.


[Diabetic polyneuropathy. 4. Synopsis of electroneurographic findings in diabetics].

Sensory conduction velocity of the median nerve, motor conduction velocity of both median and tibial nerves, and corresponding distal laterncies are sufficient parameters to establish the diagnosis of polyneuropathy almost with certainty. Considering these six parameters yielded in detection of peripheral nerve dysfunction in 22% of diabetic patients who were free from clinical signs of polyneuropathy. Electroneurographical findings in 340 out of 677 patients with diabetes mellitus were interpreted as evidence of segmental demyelination. Within this group there was the majority of patients with clinical signs of polyneuropathy and with subclinical signs of peripheral nerve dysfunction. There existed a positive correlation between signs of nerve dysfunction with angiopathy, age and duration of the disease. A second group consisting of 243 diabetics with signs of incipient segmental demyelination with or without signs of axonaal degeneration mainly included juvenile patients with a short duration of the disease and with a low frequency of angiopathy.


Painful ophthalmoplegia: the Tolosa-Hunt syndrome.

Painful ophthalmoplegia is characterized by unilateral involvement of the IIIrd, IVth and VIth cranial nerves, as well as supra- and retro-orbital pain, i.e. participation of the Vth cranial nerve. The pain is relieved within 48-72 h with steroid therapy. The paresis of the eye muscles in various combinations usually subsides gradually from within a few weeks to several months. The etiology is unknown. The few pathological examinations reported in the literature showed an unspecific inflammatory granulation tissue around the intracavernous portion of the carotid artery and on the dura mater in the vicinity of the cavernous sinus. Carotid arteriography may show stationary waves of this artery and narrowing of its intracavernous portion. With orbital phlebography the occlusion of the supraorbital vein and obstruction of the cavernous sinus are sometimes demonstrable. The syndrome is well defined and its etiology still unknown.


Electroneurographic investigations of misonidazole polyneuropathy.

13 patients with malignant tumors were treated by the radiosensitizer misonidazole (Ro 07-0582), total dosage 20-29 g. The electrophysiological investigations showed (1) an early increase of distal latency, the motor nerve conduction velocity (NCV) of the peroneal nerve and the NCV of the sural nerve remaining normal or only signlty reduced, and in a few cases a marked reduction of the compound action potential or of the nerve action potential (NAP), indicating a primary axonal neuropathy; (2) greater changes in the parameters of sensory nerves (n. suralis) than of motor nerves; (3) the distal latency is a good indicator of subclinical neuropathies; (4) the electrophysiological parameters showed a normalization 6 months after the end of therapy. The mechanisms possibly responsible for the misonidazole neuropathy are discussed.