Development of the resting sporangia ofSynchytrium endobioticum,the causal agent of potato wart disease

Summary An ultrastructural study of the development of the resting sporangium ofSynchytrium endobioticum (Schilb.) Perc. infecting potato cells is presented. The resting sporangium is found to have a single large, centrally placed nucleus with a prominent nucleolus through its entirein situ development. The cytoplasmic organization of the resting sporangium is further characterized by numerous membrane-bound lipid bodies and osmiophilic bodies. The latter have a characteristic sieve-like appearance, probably because certain storage components have been extracted during preparation for electron microscopy. Because of the similar location and appearance of these osmiophilic bodies it is suggested that they are identical to what has earlier (based on light microscopy) been described as chromatin granules; and the ultrastructural studies presented here show that nucleolar discharge which was described from light microscopic observations as leading to chromatin granules in the cytoplasm, and finally forming the nuclei of the zoospores (bally 1912,curtis 1921,percival 1910) simply does not occur.The appearance of dense fibrillar-like structures on the sporangial surface at an early stage of resting sporangium development ultrastructurally distinguishes the resting sporangium from the zoosporangium. The development of the layered portion of the thick sporangial wall is shown to be due to the fusion of vacuoles containing pre-made wall fibrils with the cell membrane. It is suggested that the inner compact wall layer which is essentially substructureless is formed by the membrane itself.The characteristic wings of the matureS. endobioticum resting sporangium originate from the potato host cell wall. Remnants of host cell organelles in the outermost layer of the resting sporangium wall show that degradation of the host cell cytoplasm contributes to wall formation of the parasite.     

The neurohypophysis of the foetal monkey

Summary The neurohypophysis of foetal macaque monkeys has been studied by optical and electron microscopy. Abundant elementary neurosecretory granules are present in the infundibular process by the middle third of gestation. Most of these are of variable electron density, and surrounded by a membrane larger than the granule, so that they appear haloed. A few fibres contain membrane-bounded electron-dense granules which show no halo. Inclusions of synaptic vesicle size are rare. The infundibular stem contains a few fibres with typical inclusions smaller than 1000 Å in diameter; the latter resemble inclusions in fibres of the median eminence.Dedicated to Professor W. Bargmann on his 60th birthday.     

Ultrastructure of light cells in the dolphin thyroid

Summary The ultrastructural appearance of the light cells is described in the thyroid gland of the common (Pacific) dolphinDelphinus bairdi. The appearances of active light cells are discussed in relation to the production of thyrocalcitonin and the special osteological characteristics of marine mammals. The presence of nerve processes possibly associated with light cells is discussed.Also known as parafollicular cells (Nonidez, 1932) and C cells (Pearse, 1966).     

Sequence analysis ofp-hydroxyphenyl-O-β-d-xyloside initiated and radio-iodinated dermatan sulfate from skin fibroblasts

To generate xyloside-primed dermatan sulfate suitable for sequence analysis, skin fibroblasts were incubated withp-hydroxyphenyl--d-xylopyranoside and [³H]galactose, and free [³H]glycosaminoglycan chains were isolated from the culture medium by ion exchange and gel chromatography. After¹²⁵I labelling of their reducing-terminal hydroxyphenyl groups, chains were subjected to various chemical and enzymatic degradations, both partial and complete, followed by gradient polyacrylamide gel electrophoresis and autoradiographic identification of fragments extending from the labelled reducing-end to the point of cleavage. Results of periodate oxidation-alkaline scission indicated that the xylose moiety remained unsubstituted at C-2/C-3; exhaustive treatment with chondroitin AC-I lyase afforded the fragment HexA-Gal-Gal-Xyl-R (R = radio-iodinated hydroxyphenyl group), and complete degradations with chondroitin ABC lyase as well as testicular hyaluronidase yielded the fragments HexA/HexA-GalNAc-GlcA-Gal-Gal-Xyl-R with or without sulfate on theN-acetylgalactosamine. Partial digestions with testicular hyaluronidase or chondroitin B lyase indicated that glucuronic acid was common in the first three repeats after the linkage region and that iduronic acid could occupy any position thereafter. Hence, there were no indications of a repeated, periodic appearance of the clustered GlcA-GalNAc repeats which was previously observed in proteoglycan derived dermatan sulfate [Fransson L-Å, Havsmark B, Silverberg I (1990)Biochem J 269:381–8], suggesting a role for the protein part in controlling the formation of particular copolymeric features during glycosaminoglycan assembly.Abbreviations GAG glycosaminoglycans - CS chondroitin sulfate - DS dermatan sulfate - Ser serine - Xyl d-xylose - Gal d-galactose - GlcA d-glucuronic acid - IdoA l-iduronic acid - GalNAc N-acetyl-d-galactosamine - GlcNAc N-acetyl-d-glucosamine - HexA 4-deoxy-l-threo-hex-4-enopyranosyluronic acid - HO-Phe p-hydroxyphenyl group - HO-Phe-Xyl p-hydroxyphenyl-O--d-xylopyranoside - O₂N-Phe-Xyl p-nitrophenyl--d-xylopyranoside - OSO₃ ester sulfate - PAGE polyacrylamide gel electrophoresis - HPLC high performance liquid chromatography - FPLC fast performance liquid chromatography - LC standard liquid chromatography     

Lectin histochemistry of complex carbohydrates in human cervix

Summary Complex carbohydrates in the human cervix were studied histochemically using lectins, conjugated to horseradish peroxidase and correlated procedures. Stratified squamous epithelium of the exocervix and columnar epithelium of the endocervix in some, but not all specimens showed staining for terminal -N-acetyl-d-galactosamine, -d-galactose, -d-galactose and -l-fucose. the staining for -N-acetylgalactosamine and -galactose, the terminal sugars in blood group A and B antigens respectively, corresponded to a large extent with ABO blood type. One exception was the lack of staining for terminal -N-acetylgalactosamine in endocervical secretions in three of nine blood type A patients. A second exception was the staining for terminal -galactose in endocervical secretions in about half of blood type O and A specimens. The type and amount of glycoprotein formed by endocervical columnar cells differed according to location in superficial compared with deep portions of the glands and according to location at the junction with exocervix compared with the more internal regions. Staining of endothelial cells for blood group A and B antigens was confined to subjects of blood type A and B respectively, although three of nine type A specimens showed no lectin reactivity for group, A antigen. Endothelial cells evidenced affinity forUlex europeus I agglutinin demonstrative of fucose in all specimens. Mast cells disclosed lectin affinity consistent with the presence of terminal or internal mannose orN-acetylglucosamine residues. Two blood type O specimens were examined with conjugated lectins at the ultrastructural level. Secretory granules stained for content of terminal -galactose, -galactose and fucose. These results support and concur with biochemical studies of complex carbohydrates in human cervical tissues. They reveal, in addition, the location of the blood group antigens in the human exocervix and endocervix and the marked heterogeneity among endocervical columnar cells in glycoprotein production.     

Effects of α-galactosidase digestion on lectin staining in human pancreas

Summary Effects of -galactosidase (from green coffee beans) digestion on lectin staining were examined in formalin-fixed, paraffin-embedded human pancreatic tissues from individuals of blood-group B and AB. Digestion with the enzyme resulted in almost complete loss of Griffonia simplicifolia agglutinin I-B₄(GSAI-B₄) staining in the acinar cells with concomitant appearance of Ulex europaeus agglutinin-I(UEA-I) staining in the corresponding cells. In addition, reactivity with soybean agglutinin(SBA) was also imparted by the enzyme digestion in GSAI-B₄ positive acinar cells. -Galactosidase digestion following -galactosidase digestion neither reduced the reactivity with SBA nor induced the reactivity with Griffonia simplicifolia agglutinin-II(GSA-II) in GSAI-B₄ positive cells, while in UEA-I positive cells, both reduction of SBA reactivity and appearance of GSA-II reactivity occurred after simple -galactosidase digestion as well as sequential digestion with - and -galactosidase. However, when -l-fucosidase digestion procedure was inserted between - and -galactosidase digestion, UEA-I staining imparted by -galactosidase digestion was markedly decreased in intensity and GSA-II reactivity was appeared in GSAI-B₄ positive acinar cells. Furthermore, after sequential digestion with -galactosidase and fucosidase, reactivity with peanut agglutinin(PNA) was revealed in GSAI-B₄ positive acinar cells as well as UEA-I positive cells in secretors. In non-secretors, strong PNA staining was usually observed in the acinar cells throughout the glands without enzyme digestion. These results confirmed that the -galactosidase induced GSA-II reactivity and the fucosidase induced PNA reactivity are due to precursors of different kinds of blood-group determinants and suggest that at least two kinds of B antigen determinants, i.e. Gal(1-3)[Fuc(1-2)]Gal(1-3,4)GlcNac and Gal(1-3)-[Fuc(1-2)]Gal(1-3)GalNAc are produced in GSAI-B₄ positive acinar cells. The synthesis of the latter type of B antigen is assumed to be controlled under the secretory gene in human pancreas.Abbreviation GalNAc N-acetyl-d-galactosamine - Gal d-galactose - GlcNAc N-acetyl-d-glucosamine - Fuc l-fucose - NeuNAc N-acetylneuraminic acid (sialic acid)     

Location by fluorescence microscopy of glycosidases and a xylanase in the anaerobic gut fungi Caecomyces communis,Neocallimastix frontalis,and Piromyces rhizinflata

-d-Glucosidase, -d-fucosidase -d-xylosidase, and -cellobiopyranosidase activities in Caecomyces communis, Neocallimastix frontalis, and Piromyces rhizinflata, located with fluorescent conjugates, occur throughout the whole thallus as from zoospore germination and disappear before sporulation. -d-Galactosidase and -l-arabinopyranosidase activities are low or nonexistent. A xylanase, detected by indirect immunofluorescence, was observed at the surface of the vegetative cells, vesicles, or rhizoids. Cross-reactions prove the existence of analogies in structure among the enzymes of these anaerobic gut fungi.     

Deletion mapping of galactose mutants with transducing lambda phages

Summary The present mapping studies in the gal region of E. coli are a continuation of experiments reported previously (Pfeifer and Oellermann, 1967).By mapping of 238 gal mutants against 145 deletions of the gal region carried by dg phages the kinase gene could be divided into 14 and the transferase gene into 19 deletion groups¹. Results of quantitative transduction experiments strongly suggest that there are preferred endpoints for the deletions.     

Elektronenmikroskopische Untersuchungen am Stirnorgan von Anuren

The ultrastructure of the frontal organ (pineal end-vesicle, Stirnorgan) of Rana temporaria L. and Rana esculenta L. is similar to the submicroscopic organization of the retina and other photosensitive organs. There are five different cell types in the frontal organ: sensory (receptor) cells, ependymal cells, ganglion cells, glial cells and epithelial cells. The ependymal cells may be secretory. There is no evidence for a typical pigment epithelium. The sensory cells have inner and outer segments. The inner segments contain numerous mitochondria, a Golgi complex, filaments, lipid droplets, two centrioles and a fibrillar apparatus (within the connecting piece). The mitochondria are very abundant in the Ellipsoid and Ersatzellipsoid areas (Holmgren) of the inner segment. The outer segment consists of about 60 to 110 discs formed by infoldings of the cell membrane. Most of the sensory cells are cone-like, but there are some elements with rod-like structures. Plexiform areas of the frontal organ contain terminations of the receptor cells, and processes of the nerve cells and glia cells. Synaptic structures have been determined within these areas. Non-medullated and medullated nerve fibers with adjacent glial satellites are observed in the pineal nerve (Nervus pinealis). The anatomical findings are described in detail and discussed in respect to the physiological results of Dodt and Heerd (1962) in Rana temporaria and Rana esculenta.

---

---

Durchgeführt mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Design of superactive and selective integrin receptor antagonists containing the RGD sequence

Summary Integrins play a major role in cell-cell and cell-matrix interactions. The majority of the different types of integrins recognize the tripeptide sequence arginine-glycine-aspartic acid (RGD). To explore the spatial requirements of the pharmacophore for receptor selectivity and high activity, a new procedure, spatial screening, was used. The procedure is based on the experience that the conformation of small cyclic peptides is mainly determined by the chirality of the amino acids (and glycine or proline). For example, cyclic pentapeptides with one d and four l amino acids prefer a II'/ conformation. The sequence RGDFV was shifted around this spatial II'/ template by synthesis of five peptides in which one of the amino acids was used in d-configuration. It turned out that cyclo(-RGDfV-) is a selective inhibitor for the _v3 integrin, which is strongly expressed in cancer cells. Systematic variations with different turn mimetics, retro-inverso structures, modified peptide bonds and sugar amino acids are discussed.     

On the gas vacuoles of the halobacteria

Summary The cells of Halobacterium sp., strain 5, contain a large number of highly refractile bodies of the type which Petter (1932) suggested were gas-filled vacuoles. The present studies support Petter's contention, but the evidence for the exact chemical nature of the vacuole content is still indirect. It is not carbon dioxide or oxygen, but might possibly be nitrogen. Strain 5 loses spontaneously and with a high frequency the ability to make the vacuoles.When vacuolated cells are subjected to pressure, the vacuoles disappear, but can recover upon aeration. Oxygen and the organic constituents of the growth medium stimulate the recovery, whereas 2.4-dinitrophenol inhibits it. A procedure is described for the isolation of the vacuoles. The vacuoles are bounded by a membrane which reveals itself in electron micrographs of thin sections as a 1-layered structure about 30 Å thick.Dedicated to Prof. C. B. van Niel on the occasion of his 70th birthday.     

Maturation of black spruce somatic embryos. Part I. Effect of l-glutamine on the number and germinability of somatic embryos

Different concentrations of l-glutamine and different nitrogen sources in the medium were compared during maturation of black spruce (Picea mariana (Mill.) B.S.P.) somatic embryos. l-glutamine can be used as the sole nitrogen source for the maturation of Picea mariana somatic embryos at 2 to 3 gl^-1. A significantly lower number of somatic embryos was obtained on a medium prepared with only inorganic nitrogen. Compared with a medium supplement to inorganic nitrogen resulted in a twofold increase in the number of embryos for six genotypes. The nitrogen source and concentration in the maturation medium significantly affected the germination sensus stricto of somatic embryos (radicle appearance), but not their development into plantlets; at the time of epicotyl appearance, an effect of the nitrogen source was no longer found. A comparison of the development of somatic embryos into plantlets from seven genotypes showed that the genotype had more effect in terms of epicotyl appearance and in conversion rate than the nitrogen source present in the maturation medium.Abbreviations HLM-1 half-Litvays's medium with 10 M 2,4-D and 5 M BA - i only inorganic nitrogen in the medium - i+1 gG inorganic nitrogen plus 1 g l^-1 glutamine in the medium - SMM standard maturation medium - 2.5gG only 2.5 g l^-1 glutamine in the medium     

Die dünnen und die dicken Muskelfasern des Zwerchfells der Ratte

Zusammenfassung Einleitend wird auf die vielfältigen, z.T. sich widersprechenden Resultate über die gefelderten und fibrillären Muskelfasern der Wirbeltiere hingewiesen.In den Befunden wird gezeigt, daß in den dünnen Muskelfasern des Zwerchfells der Ratte in I gelegene Mitochondrienroste durch longitudinale Verbindungen zu einem Gerüst verbunden sind. Die Mitochondrien der dicken Fasern beschränken sich auf unverbundene Netze in den isotropen Schichten der Sarkomere. Der prozentuale Anteil der Mitochondrien am Gesamtfaservolumen ist bei den dicken Fasern bis zur Hälfte geringer als bei den dünnen. Außerdem erhalten sie weniger Glykogen und kein Fett in Tropfenform.Anhand der Abstände der Myosinfäden wird der Mindestabstand der in Z quadratisch angeordneten Aktinfilamente berechnet.Die unterschiedlichen Formen des T-Systems in kontrahiertem und gedehntem Zustand der Faser sind Anlaß, die an Membranmodellen gewonnenen Vorstellungen über die Membraneigenschaften auf die Triaden zu übertragen.Die verschiedenen Anteile des interfibrillären Raumes werden zu den sie umgebenden Strukturen in funktionelle Beziehung gesetzt.

---

Summary Existing information on mammalian muscle fibers with Fibrillenstruktur and of those with Felderstruktur (Krüger) is in part contradictory. The present study reveals in the thin fibers (Felderstruktur) of the rat diaphragm, mitochondrial grids at the level of each I-disc. These are joined by means of longitudinal connections to form a continuous three-dimensional mitochondrial framework. In the thick fibers (Fibrillenstruktur) the mitochondria form only discontinuous nets or sieves in each isotropic layer of the sarcomere. In thick fibers the amount of mitochondrial material relative to total fiber volume is only about 1/2 of that in thin fibers. Furthermore, the thick fibers contain less glycogen and no lipid in the form of droplets. The T-systems differ in appearance in the contracted and in the expanded state of the fiber. This suggests the extension of the concepts on properties of membranes derived from membrane models to include the triads. The various components of the interfibrillar space are brought into functional relationship to the acjacent structures.

---

---

Herrn Professor Dr. W. Bargmann zum 60. Geburtstag gewidmet.     

Effects of divalent cations on toad end-plate channels

Summary Miniature end-plate currents (MEPCs) and acetylcholine-induced current fluctuations were recorded in voltageclamped, glycerol-treated toad sartorius muscle fibers in control solution and in solutions with added divalent cations. In isosmotic solutions containing 20mm Ca or Mg, MEPCs had time constants of decay ( _D ) which were about 30% slower than normal. In isotonic Ca solutions (Na-free), greater increases in both _D and channel lifetime were seen; the null potential was –34 mV, and single-channel conductance decreased to approximately 5 pS. Zn or Ni, at concentrations of 0.1–5mm, were much more effective in increasing _D than Ca or Mg, although they did not greatly affect channel conductance. The normal temperature and voltage sensitivity of was not significantly altered by any of the added divalent cations. Surface potential shifts arising from screening of membrane fixed charge by divalent cations cannot entirely explain the observed increases in , especially when taken together with changes in channel conductance.     

Induction of extracellular arabinases on monomeric substrates in Aspergillus niger

The induction of extracellular arabinases by pentose sugars and polyols generated by the metabolic pathway of l-arabinose and d-xylose catabolism in Aspergillus niger was investigated. Induction occurred with l-arabinose and l-arabitol but not with d-xylose or xylitol. l-arabitol in particular was found to be a good inducer for -l-arabinofuranosidase and endo-arabinase activities. Western blotting analysis showed both -l-arabinofuranosidase A and B to be present. No induction was observed using d-arabitol. Unlike the wild type A. niger N402 strain, the A. niger xylulose kinase negative mutant N572 also showed induction of -l-arabinofuranosidases A and B and endo-arabinase activity on d-xylose and xylitol. This is due to metabolic conversion of these compounds leading to the accumulation of both xylitol and l-arabitol in this mutant, the latter of which then acts as inducer. The induction of the two -l-arabinofuranosidases and endo-arabinase is under the control of two regulatory systems namely pathway specific induction and carbon catabolite repression. Under derepressing conditions in the wild type only -l-arabinofuranosidase B could be detected by Western blotting analysis. This indicates that -l-arabinofuranosidase B is of importance in the initiation of specific induction of the various arabinose activities in A. niger grown on arabinose containing structural polysaccharides.Abbreviations PNA p-nitrophenyl--l-arabinofuranoside     

Properties of Aspergillus japonicus β-fructofuranosidase immobilized on porous silica

-Fructofuranosidase from Aspergillus japonicus MU-2, which produces fructo-oligosaccharides (1-kestose: O--D-fructofuranosyl-(2 1)--D-fructofuranosyl -D-glucopyranoside); and nystose: O--D-fructofuranosyl-(2 1)--D-fructofuranosyl-(2 1)--D-fructofuranosyl -D-glucopyranoside) from sucrose, was immobilized, covalently with glutaraldehyde onto alkylamine porous silica, at high efficiency (64%). Optimum pore diameter of porous silica for immobilization of the enzyme was 91.7 nm. After immobilization, the enzyme's stabilities to temperature, metal ions and proteolysis were improved, while its optimum pH and temperature were unchanged. The highest efficiency of continuous production of fructo-oligosaccharides (more than 60%), using a column packed with the immobilized enzyme, was obtained at 40% to 50% (w/v) sucrose. The half-life of the column during long-term continuous operation at 55°C was 29 days.     

Capillarization,mitochondrial densities,oxygen diffusion distances and innervation of red and white muscle of the lizard Dipsosaurus dorsalis

Summary The white and red regions of the iliofibularis muscle of the lizard Dipsosaurus dorsalis were analyzed using histologic and morphometric analysis. These regions are composed of fast glycolytic (FG) and both fast oxidative, glycolytic (FOG) and tonic fibers, respectively. Endplate morphology and number of endplates per fiber were estimated from fibers from both areas. Capillary volume densities of the red and white regions were quantified from transverse sections. Mitochondrial volume of fibers from the red and white regions were estimated from electron micrographs.All fibers from the white region of the iliofibularis possessed a single, well defined endplate, as did most red region fibers. The remaining red fibers (28±5%) possessed an average of 14.7±3 endplates each, distributed along the entire length of the fiber at intervals of approximately 1124 m.Red fibers possessed twice the mitochondrial volume of white fibers (7.6±0.4%, red; 3.8±0.3%, white). Mitochondria were distributed uniformly through the fibers from both regions. Capillary anisotropy was low ( = 1.018) in both regions. Capillary densities of the red region (629±35 mm^-2) were much greater than those of the corresponding White region (73±8 mm^-2).The data indicate that capillary densities, mitochondrial volumes and theoretical diffusion distances correlate well with the oxidative capacity of lizard muscle fibers. Tonic fibrs of this species appear oxidative and therefore metabolically capable of functioning during locomotion. The similar mitochondrial volumes and capillary densities of reptilian and mammalian muscles suggest that the greater oxidative capacity of mammalian muscle is due in part to possession of more oxidatively active mitochondria rather than to possession of more mitochondria per se.     

Gibberellins play a role in the interaction between imidazole fungicides and cytokinins in Araceae

Imidazole fungicides such as imazalil, prochloraz, and triflurnizole and the triazole growth retardant paclobutrazol promote the shoot-inducing effect of exogenous cytokinins in Araceae, such as Spathiphyllum floribundum Schott and Anthurium andreanum Schott. The mechanism of their action could partially be based on the inhibition of gibberellic acid (GA) biosynthesis, because administration of GA₃ inhibits the phenomenon completely in S. floribundum. Not only is the suppression of GA biosynthesis involved, but also the metabolism of endogenous cytokinins is significantly altered. Although the balance between isopentenyladenine, zeatin, dihydrozeatin, and their derivatives was shifted to distinguished directions by administration of BA and/or imazalil and/or GA₃, no correlation between these changes in metabolic pathways and the number of shoots could be found. The metabolism of BA was not significantly altered by adding imazalil to the micropropagation medium of S. floribundum.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - [9R-5P]DHZ 9--d-ribofuranosyl-dihydrozeatin-monophosphate - [9R-5P]iP 6-isopentenyl-9--d-ribofuranosyladenine-monophosphate - [9R-5P]Z 9--d-ribofuranosyl-zeatin-monophosphate - [9G]BA 6-benzyl-9--d-glucopyranosyladenine - [9G]DHZ 9--d-glucopyranosyl-dihydrozeatin - [9G]iP 6-isopentenyl-9--d-glucopyranosyladenine - [9G]Z 9--d-glucopyranosyl-zeatin - [9R]BA 6-benzyl-9--d-ribofuranosyladenine - [9R]DHZ 9--d-ribofuranosyl-dihydrozeatin - [9R]iP 6-isopentenyl-9--d-ribofuranosyladenine - [9R]Z 9--d-ribofuranosyl-zeatin - BA 6-benzyladenine - DHZ dihydrozeatin - ES⁺ LC-MS/MS HPLC coupled Electrospray Tandem Mass Spectrometry - f.m. fresh mass - mT 6-(3-hydroxybenzyl)adenine - IMA imazalil - iP isopentenyladenine - NAA 1-naphthalene acetic acid - NFT Nutrient Film Technique - (OG)[9R]DHZ O--glucopyranosyl-9--d-ribofuranosyl-dihydrozeatin - (OG)[9R]Z O--d-glucopyranosyl-9--d-ribofuranosyl-zeatin - (OG)DHZ O--d-glucopyranosyl-dihydrozeatin - (OG)Z O--d-glucopyranosyl-zeatin - PAR Photosynthetic Active Radiation - PBZ paclobutrazol - PRO prochloraz - TDZ thidiazuron - TRI triflurnizole - Z zeatin     

Action of phenazine methyl sulfate,inhibitors, and uncouplers on the light-induced proton transport by cells ofRhodospirillum rubrum

Summary Suspensions of log phase cells ofRhodospirillum rubrum at pH 5.5 show a light-induced decrease in the pH of the medium which is reversed during the subsequent dark period. The velocity and magnitude of the pH change were the same whether the cells were bubbled with air, CO₂-free air or N₂ during experimentation. The pH response is temperature dependent. Phenazine methyl sulfate (PMS) at concentrations above 0.05mm stimulates the light-induced pH change. PMS at 1mm gives a 2-fold increase in the initial rate upon illumination and a 1.5-fold increase in the total change in pH after 2 min of illumination. The inhibition of the proton transport by 10 g/ml antimycin A or 20 m 2-n-heptyl-4-hydroxyquinoline-N-oxide can be partially relieved by PMS. However, inhibition of the light-induced proton transport with 0.5mm 2,4-dinitrophenol or 3 m carbonylcyanide-m-chlorophenylhydrazone (CCCP) cannot be overcome by addition of PMS. Valinomycin, at a concentration of 3 m, caused a slight stimulation of the light-induced proton transport in the presence of 200mm KCl. The inhibition of proton transport by 3 m CCCP was partially relieved with 3 m valinomycin in the presence of 200mm KCl, but the antibiotic was without effect when the cells were suspended in 200mm NaCl. The results are discussed in terms of current theories of the action of PMS, antimycin A, valinomycin, and uncouplers on the light-induced electron flow and photophosphorylation inR. rubrum.     

Purification and characterization of α(2-6)-sialyltransferase from human liver

A Gal1-4GlcNAc (2-6)-sialyltransferase from human liver was purified 34 340-fold with 18% yield by dye chromatography on Cibacron Blue F3GA and cation exchange FPLC. The enzyme preparation was free of other sialyltransferases. It did not contain CMP-NeuAc hydrolase, protease, or sialidase activity, and was stable at –20°C for at least eight months. The donor substrate specificity was examined with CMP-NeuAc analogues modified at C-5 or C-9 of theN-acetylneuraminic acid moiety. Affinity of the human enzyme for parent CMP-NeuAc and each CMP-NeuAc analogue was substantially higher than the corresponding Gal1-4GlcNAc (2-6)-sialyltransferase from rat liver.Abbreviations FPLC fast protein liquid chromatography - NeuAc 5-N-acetyl-d-neuraminic acid - 9-amino-NeuAc 5-acetamido-9-amino-3,5,9-trideoxy-d-glycero-2-nonulosonic acid - 9-acetamido-NeuAc 5,9-diacetamido-3,5,9-trideoxy-d-glycero--d-2-nonulosonic acid - 9-benzamido-NeuAc 5-acetamido-9-benzamido-3,5,9-trideoxy-d-glycero--d-galacto-2-nonulosonic acid - 9-fluoresceinyl-NeuAc 9-fluoresceinylthioureido-NeuAc - 5-formyl-Neu 5-formyl--d-neuraminic acid - 5-aminoacetyl-Neu 5-aminoacetyl--d-neuraminic acid - CMP-NeuAc cytidine-5-monophospho-N-acetylneuraminic acid - G_M1 Gal1-3GalNAc1-4(NeuAc2-3)Gal1-4Glc-ceramide - ST sialyltransferase - DTE 1,4-dithioerythritol Enzyme: Gal1-4GlcNAc (2-6)-sialyltransferase, EC 2.4.99.1.