Synthesis and stability of arylsulfatase A and B in fibroblasts from multiple sulfatase deficiency

Fibroblasts from patients with multiple sulfatase deficiency were analyzed for activities of arylsulfatase A and B, iduronate 2-sulfatase and sulfamatase. A group of patients (group I) severely deficient in all sulfatases (residual activities less than or equal to 10% of control) were differentiated from patients (group II) with residual sulfatase activities of up to 90% of control. The synthesis and stability of arylsulfatase A and B were determined in pulse-chase labelling experiments. The apparent rate of synthesis of arylsulfatase A and B varied from 30% to normal in both fibroblasts from group I and II multiple sulfatase deficiency. In group I the molecular activity of the arylsulfatase A and B was more than 10-fold lower than in control fibroblasts. In group II the molecular activity of the arylsulfatase A was twofold to threefold lower and that of arylsulfatase B half of normal. In fibroblasts of both groups the stability of arylsulfatase A polypeptides was significantly diminished. For arylsulfatase B the instability was restricted to the mature 47000-Mr polypeptide and was variable within both groups. These results demonstrate that multiple sulfatase deficiency is a heterogeneous disorder, in which the primary defects can impair both the catalytic properties and the stability of sulfatases.     

Gamma-radiolysis of N2O-saturated formate solutions. A chain reaction

In the radiolysis of aqueous formate-containing solutions a chain reaction (i, ii) proceeds in the presence of N2O. CO2-. + N2O + H2O----CO2 + N2 + .OH + OH- (i) .OH + HCO2-.----CO2-. + H2O (ii) The chain length depends on the dose rate and the N2O concentration but not on the formate concentration. Typically, G(CO2) approximately 140 molecules (100 eV)-1 is found, with an equivalent amount of N2, at a dose rate of 3 X 10(-3) Gy s-1. The rate constant for the rate-determining step in this chain reaction has been calculated at k(i) = 1600 dm3 mol-1 s-1. The possible relevance of this chain reaction in radiation biological studies is briefly discussed.     

Production of molecular hydrogen with immobilized cells ofRhodospirillum rubrum

Summary Cells ofRhodospirillum rubrum have been immobilized in various gels and tested for photobiological hydrogen production. Agar proved to be the best immobilizing agent with respect to production rates as well as stability. Agar immobilized cells were also superior compared to liquid suspension cultures. Growth conditions of the cells prior to immobilization, e.g. cell age, light intensity or nutrient composition, were of primary importance for the activity in the later immobilized state. A reactor with agar immobilized cells has been operated successfully over 3000 h with a loss of the activity of about 60%. Mean rates for hydrogen production for immobilized cells in this work during the first 60 to 70 hours after immobilization were in the range of 18 to 34 μl H₂ mg⁻¹ d.w. h⁻¹ and thus by a factor of up to 2 higher than liquid cultures under the same conditions. Maximal rates of hydrogen production (57 μl H₂ ml⁻¹ immobilized cell suspension) were reached in agar gel beads with cells immobilized after 70 h growth in liquid culture in the light and a cell density of 1.0 mg ml⁻¹, 70 h after immobilization.     

Cell surface influenza haemagglutinin can mediate infection by other animal viruses.

We have used filter-grown Madin-Darby canine kidney (MDCK) cells to explore the mechanism by which influenza virus facilitates secondary virus infection. Vesicular stomatitis virus (VSV) and Semliki Forest virus (SFV) infect only through the basolateral surface of these polarized epithelial cells and not through the apical surface. Prior infection with influenza virus rendered the cell susceptible to infection by VSV or SFV through either surface. The presence of both a permissive and a restrictive surface for virus entry in the same cell allowed us to determine how the influenza infection enhanced the subsequent infection of a second virus. Biochemical and morphological evidence showed that influenza haemagglutinin on the apical surface serves as a receptor for the superinfecting virus by binding to its sialic acid-bearing envelope proteins. Influenza virus also facilitates secondary virus infection in non-epithelial cells; baby hamster kidney cells (BHK-21), which are normally resistant to infection by the coronavirus (mouse hepatitis virus MHV-A59), could be infected via the haemagglutinin-sialic acid interaction. Facilitation of secondary virus infection requires only the sialic acid-binding properties of the haemagglutinin since the uncleaved haemagglutinin could also mediate virus entry.     

The development of the human spleen

Summary Splenic tissue of human fetuses from the 14^th to the 24^th week of gestation (menstrual age) were investigated by light- and electron microscopy to describe the development of the red and white pulp in close relationship to the differentiation of the vascular tree. Special interest is focussed on the differentiation of the T-cell- and the B-cell regions and their specific stationary cells.The preliminary stage, here called the primary vascular reticulum, lasts up to the 14^th gestational week (gw). Numerous erythrocytes, normoblasts and macrophages are seen among a network of mesenchymal cells and argyrophilic fibers. Hematopoiesis, especially erythropoiesis, can be recognized.The characteristic organ structure becomes established during the subsequent transformation stage of the fetal spleen, beginning with the 15^th gw. Splenic lobules begin to form during the 15^th to 17^th gw. They consist of a central artery, surrounded by a sheath of lightly stained stationary cells which resemble myofibroblasts. At the periphery of these lobules the red pulp forms. Initially mobile cells are distributed throughout the reticulum. Soon they begin to accumulate in the venous sinuses, which develop from lacunae among the reticular network and come into contact with the venous system. The endothelial wall of these sinuses remains discontinuous, confirming the theory of the open vascularization of the spleen. The development of the larger veins is correlated with the differentiation of the splenic trabeculae.The development of the white pulp is correlated with the stage of lymphoid colonization within the spleen, beginning around the 18^th gw. An accumulation of lymphocytes around the central arteries can be recognized during the 19^th and 20^th gw. These lymphoid cells show morphological and immunohistochemical characteristics of T-precursor cells. Within the now assembling periarterial lymphoid sheath (PALS) a few precursors of interdigitating cells (IDC) are recognizable, giving evidence for the differentiation of the T-cell region.Around the 23^rd gw the assemblage of primary follicles is discernible at the periphery of the PALS. Precursors of the follicular dendritic reticulum cell (FDRC), the specific stationary cell of the B-cell region, have been recognized. This observation leads to the conclusion that the small primary follicles represent the beginning formation of the B-cell region.The significance of the vascular system for the differentiation of the specific splenic organization is discussed.This investigation was supported by grants from the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 111)The authors appreciate the contribution of human fetal material from Dr. von Hollweg and Dr. Körner from the Hospital Heidberg, Hamburg, and the excellent technical assistance of Mrs. H. Hansen, Mrs. I. Knauer, Mrs. M.v. Kolszynski, Mrs. J. Quitzau, Mrs. H. Siebke and Mrs. H. Waluk     

Metabolic,physiological and kinetic aspects of the alcoholic fermentation of whey permeate by Kluyveromyces fragilis NRRL 665 and Kluyveromyces lactis NCYC 571

Optimum growth conditions for the fermentation of non-concentrated whey permeate by Kluyveromyces fragilis NRRL 665 have been defined. Use of 3.75 g yeast extract l^?1, a growth temperature of 38°C and a pH of 4.0 allowed a maximum productivity of 5.23 g ethanol l^?1 h^?1 in continuous culture with a yield 91% of theoretical. Complete batch fermentation of permeate with 100 g lactose l^?1 was possible with a maximum specific growth rate of 0.276 h^?1 without any change in ethanol yield. Fermentation of concentrated permeate resulted, however, in a general decrease of specific substrate consumption rate, demonstrated by the inability to completely convert an initial 90 or 150 g lactose l^?1 in continuous culture, even at dilution rates as low as 0.05 and 0.08 h^?1, respectively. The decrease could be related to substrate inhibition, to an increase in osmotic pressure caused by lactose and salts, and to ethanol inhibition of both alcohol and biomass yield. The decrease in specific productivity could be counterbalanced by use of high cell density cultures, obtained by cell recycle of K. fragilis. Fermentation of a non-concentrated permeáte at a dilution rate of 1 h^?1 resulted in a productivity of 22 g l^?1 h^?1 at 22 g ethanol l^?1. Cell recycle using flocculating Kluyveromyces lactis NCYC 571 was also tested. With this strain a productivity of 9.3 g l^?1 h^?1 at 45 g product l^?1 was attained at a dilution rate of 0.2 h^?1, with an initial lactose concentration of 95 g l^?1.     

Differentiation patterns in the Scutellaria albida group (Lamiaceae) in the Aegean area

An experimental investigation of the mainly white-flowered Scutellaria albida group in the Aegean area was carried out. Eight populations representing maximum morphological variation as well as geographic separation were chosen for crosses between populations. Strong sterility barriers were observed in the Cretean endemic species, S. sieberi Benth., which also deviated morphologically and is probably an ancient relic. Strong crossing barriers were also found in plants from Euboea, and, in spite of little morphological differentiation from S. albida L. s. str., they are treated as a separate species, S. goulimyi Rech. fil. Crossing barriers of intermediate strength were present in morphologically distinct material from N Sporades, which is also treated as a separate species, S. sporadum Bothmer sp. nov. All other populations showed high compatibility in crosses among themselves, viz. populations from Bulgaria, Thrace, Andros, Naxos, and Rodhos. These were treated as the same species, S. albida , having large local as well as clinal variation. Three subspecies are retained: ssp. albida in the north; ssp. perhispida (Bornm.) Bothmer in E Macedonia, Thrace and on the islands; and the mainly purple-flowered, ssp. vacillans (Rech. fil.) Bothmer, endemic to the peninsula of Athos. Evolutionary pathways and phytogeographical patterns are discussed.
S. sporadum is mainly inbreeding and, in S. sieberi , indications of a self-incompatibility system were found. The other taxa have a more versatile system.     

Internalization of blocking antibodies against mannose-6-phosphate specific receptors.

Antibodies against mannose-6-phosphate specific receptors inhibit the receptor-dependent endocytosis of exogenous lysosomal enzymes as well as the sorting of endogenous lysosomal enzymes. This inhibition was correlated with an apparent loss of the receptors. We report here that treatment of cells with the antibody results in the formation of receptor-antibody complexes that are not extracted by the procedure used for the solubilization of receptors prior to immunoprecipitation and detection of the receptor. The apparent loss of receptors is observed with both native antibody and the F(ab)2 fragments, but not with Fab fragments. In contrast the transport of lysosomal enzymes is inhibited by all three forms of the antibody. The inhibition is ascribed to masking by the antibody of the enzyme-binding site in the receptor. The inhibition of the sorting of endogenous lysosomal enzymes by antibodies added to the medium indicates that the mannose-6-phosphate specific receptors at the sorting site are in dynamic equilibrium with those at the cell surface. The receptor-antibody complexes formed at the cell surface appear to cycle between the cell surface and intracellular membranes. A fraction of the internalized antibodies dissociates from the receptors and is degraded after transfer into lysosomes. Complexing with Fab increases the concentration of the receptor in the lysosomes and decreases 2- to 3-fold the half-life of the receptor.     

Effects of dehydration on the vasopressin response to immersion

Nine healthy volunteers underwent three experimental procedures in random order. The protocols were 4 h of thermal dehydration followed by 2 h of head-out water immersion, 4 h of thermal dehydration followed by 2 h of chair rest, and 6 h of rest in the supine position. Four hours of heat exposure (50 degrees C) resulted in a body weight loss of approximately 3.5%. Plasma osmolality rose by approximately 5 mosmol/kg, mean arterial pressure (MAP) decreased from 85 to 78 mmHg, and body temperature increased from 36.8 to 38.6 degrees C. As a consequence of the combined action of hypertonicity, hypovolemia, hypotension, and hyperthermia, plasma arginine vasopressin (AVP) increased from 2.1 to 8.1 pg/ml after 4 h thermal dehydration. Changes in body weight, plasma osmolality, body temperature, and MAP were similar after either a subsequent 2 h of water immersion or 2 h of chair rest. However, during chair rest plasma AVP remained elevated (8.4 pg/ml), whereas during immersion plasma AVP decreased from 8.1 to 4.7 pg/ml. This was probably due to the central hypervolemia induced by immersion. Our results support the hypothesis that central hypervolemia rather than hypotonicity is the primary stimulus for AVP suppression during water immersion in dehydrated subjects. During the early immersion period hypoosmolality might contribute to the AVP suppression.