Protracted release of the LHRH agonist avorelin (MF 6001) from two depot formulations in dogs and men

Avorelin is a new superagonist of naturalluteinizing-hormone-releasing-hormone. Avorelin hasbeen formulated in high molecular weight polylactic glycolic acid to afford protracted andcontinuous release of the peptide from subcutaneousimplants. Two different formulations (10 and 15 mg)were tested first in dogs and then in men during aclinical phase II trial. Chemical castration wasmaintained for at least 6 months in dogs withboth formulations. A similar duration of activity(approximately 6 months) was observed in men.     

Nonreductive release of O-linked oligosaccharides from mucin glycoproteins for structure/function assignments as neoglycolipids: application in the detection of novel ligands for E-selectin

The neoglycolipid technology comprises several microproceduresinvolving the generation of lipid-linked oligosaccharide probesfor carbohydrate recognition studies in conjunction with oligosaccharidesequence determination by mass spectrometry. Although applicableto any desired oilgosaccharides, procedures are greatly facilitatedif the ohgosaccharides are nonreduced, as conjugation is byreductive amination of a reducing end aldehyde to a phosphatidylethanolamine.Using bovine submaxillary mucin as a model for release of O-glycansin the reducing state, and based on yields of neoglycolipidsand side-products from "peeling" reactions and degradation,aqueous ethylamine 70% w/v at 22°C for 48 h has been selectedin preference to other conditions, triethylainine, sodium hydroxide,and bydrazine. The integrity of the main acidic and neutraloligosaccharides released under these conditions, di- to octasaccharides,was established by analyses of free oligosaccharides by liquidsecondary ion mass spectrometry (LSIMS) and of the derived neoglycolipidsby TLCLSIMS; the repertoire compared favorably with that ofthe oligosaccharide alditols generated by conventional reductivealkaline borohydride treatment. More forcing conditions of ethylamine70% w/v at 65°C for 6 h were required to release oligosaccharidesfrom porcine gastric mucin; di- to nonasaccharides were obtainedof which about one-third had an intact core GalNAc. Relativeto yields after reductive alkaline hydrolysis, the overall yieldsfor these two glycoproteins were 20% and 40–50% for acidicand neutral oligosaccharides, respectively. Among O-glycansreleased from an ovarian cystadenoma glycoprotein using ethylamine,three variants of the sulfated Le^a/x sequences were identifiedas ligands for the endothelial adhesion molecule E-selectin,one of which is based on the unusual backbone Gal-3/4GlcNAc-3Gal-3Gal. mucins O-linked oligosaccharides TLC-LSIMS neoglycolipids E-selectin     

诺卡氏菌对油酸的羟基化作用

诺卡氏菌(Nocardia sp．N13)休止细胞对油酸(顾-9-十八碳烯酸)进行羟基化作用。用休止细胞转化时最适温度为30℃,最适pH为6．5,当菌体(干菌)与底物的最适比例为20 ：1对,对油酸的转化率为83．4％,不同的表面活性剂影响转化的效果。N13休止细胞在磷酸缓冲液中重复转化3次仍可保持50％左右的转化率。红外光谱、质谱、棱磁共振鉴定产物为10－羟基硬脂酸。     

Effects of actinomycin D,cycloheximide and kinetin on ribonuclease and beta-fructofuranosidase in water stressed tomato cotyledons

Three days old excised tomato cotyledons were subjected to mannitol induced water stress in the presence of actinomycin D and cycloheximide. Within a few hours, in the presence of actinomycin D but not cycloheximide, water stress induced increase in ribonuclease activities and decrease in beta-fructofuranosidase activities. The water stress action in the presence of actinomycin D was reversible by addition of kinetin. It was postulated that water stress had some immediate fundamental action on the protein synthesis sites at the ribosomes.     

Molecular forms of acetylcholinesterase present in the white and grey matter of pig brain

Extraction of the white matter of pig brain with EDTA, lysolecithin or Triton X-100 gave poor yields of soluble acetylcholinesterase although these agents had proved effective at solubilizing the enzyme in the grey matter. This finding, together with the observation that the strong detergent sodium deoxycholate, was needed to solubilize the enzyme, shows that it is more difficult to remove acetylcholinesterase from the white matter of brain than from the grey. This could mean that the enzyme in the white matter is more firmly bound to the membrane than the enzyme in the grey matter.The difference in binding of the enzyme from the two regions of the brain is also reflected in the affinity chromatography experiments which showed a lower recovery for the acetylcholinesterase of white matter compared with the enzyme from grey matter.Starch-block electrophoresis of acetylcholinesterase showed a single negatively charged peak of activity for both the naturally soluble and the deoxycholate solubilized preparations. The presence of only one form on electrophoresis suggests that the molecular species of acetylcholinesterase do not arise from differences in charge.Sucrose density gradient centrifugation of the two preparations from white matter gave a single peak of activity with a sedimentation constant of about 10 S. This corresponds closely to the major species of molecular weight 260,000 detected by gradient gel electrophoresis. Other forms detected in both enzyme preparations by gradient gel electrophoresis were species with molecular weights of 660,000, 180,000, 130,000 and 115,000. The significance of these species in terms of the formation of oligomers is discussed.A comparison was made with the corresponding preparations of acetylcholinesterase from the grey matter and the results showed that acetylcholinesterase from the white and grey matter of pig brain were very similar. The exception to this was the species with a molecular weight of 68,000 which was present in the grey but not the white matter of pig brain.     

Chromatic properties of interneurons in the optic lobes of the bee

Summary The chromatic properties of single units in the optic medulla and lobula of the worker bee were examined. This paper describes the spectral sensitivity, S () and the receptive fields of broad band units, ie. those neurons which receive qualitatively similar inputs from 2 or 3 colour receptor types.The simplest broad band unit responds with sustained excitation or inhibition to light of all colours. Intracellular staining has identified the sustained excitatory unit as the Y8 cell of the proximal medulla.More complex broad band units may receive a variety of colour inputs which sum with different weighting factors or the colour inputs may have different temporal patterning.Receptive fields tend to be large (diameter greater than 60°). The simplest broad band units show homogenous receptive fields which are uniform for all colours. More complex receptive fields contain different areas where different colours evoke an optimal response. No centre-sourround spatial antagonism was found.This work was supported by DFG grant no. Me 365/4 while J.K. was on an Alexander v. Humboldt Stipendium.We thank Joy Nelson for assistance with histology.     

Use of the spot, plate and suspension test systems for the detection of the mutagenicity of environmental agents and chemical carcinogens in Neurospora crassa.

N23 and N24, selected from hundreds of ad-3 mutants, have been used as testers for the spot, plate and suspension tests in Neurospora crassa. These two testers are highly sensitive to mutagens and are revertible by a specific group of chemicals. N23 can be reverted from an adenine-dependence to adenine-independence by agents which cause base-pair substitutions whereas N24 can be reverted by frameshift mutagens. Studies described here show that spot, plate and suspension tests using testers N23 and N24 are satisfactory substitutes for the ad-3 forward-mutation system for screening the mutagenic activity of environmental agents and chemical carcinogens in N. crassa.     

Regulation of Noradrenaline Release from Rat Occipital Cortex Tissue Chops by α2-Adrenergic Agonists

Noradrenaline (NA) and the alpha 2-adrenergic agonists clonidine, BHT-920, and UK 14304-18 inhibit potassium-evoked release of [3H]NA from rat occipital cortex tissue chops with similar potencies. NA (10(-5) M) was most effective as up to 85% inhibition could be observed compared with 75%, 55%, and 35% for UK 14304-18, clonidine, and BHT-920, respectively, all at 10(-5) M. Potassium-evoked release was enhanced by both forskolin (10(-5) M) and 1 mM dibutyryl cyclic AMP. Pretreatment of tissue chops with 1 mM dibutyryl cyclic AMP in the presence of 3-isobutyl-1-methylxanthine partially reversed the alpha 2-adrenergic agonist inhibition of NA release. No reversal of inhibition was observed following pretreatment with 10(-5) M forskolin. The effects of clonidine, BHT-920, UK-14308-18, and NA on cyclic AMP formation stimulated by (a) forskolin, (b) isoprenaline, (c) adenosine, (d) potassium, and (e) NA were examined. Only cAMP formation stimulated by NA was inhibited by these alpha 2-adrenergic agonists. These results suggest that only a small fraction of adenylate cyclase in rat occipital cortex is coupled to alpha 2-adrenergic receptors. These results are discussed in relation to recent findings that several alpha 2-adrenergic receptor subtypes occur, not all of which are coupled to the inhibition of adenylate cyclase, and that alpha 2-adrenergic receptors inhibit NA release in rat occipital cortex by a mechanism that does not involve decreasing cyclic AMP levels.     

Specificity of lung surfactant protein SP-A for both the carbohydrate and the lipid moieties of certain neutral glycolipids.

Binding specificity of the major surfactant protein SP-A from human and dog lung has been investigated. Radiobinding experiments have shown that both proteins bind in a Ca(2+)-dependent manner to galactose, mannose, fucose, and glucose linked to bovine serum albumin. These results are in accord with a previous study in which monosaccharides were linked to agarose (Haagsman, H. P., Hawgood, S., Sargeant, T., Buckley, D., White, R. T., Drickamer, K., and Benson, B. J. (1987) J. Biol. Chem. 262, 13877-13880). Chromatogram overlays in conjunction with in situ liquid secondary ion mass spectrometry (TLC-LSIMS) of several purified glycosphingolipids and neoglycolipids as well as binding assays with glycolipids immobilized on plastic wells, demonstrate recognition of galactose (human and dog SP-A), glucose, and lactose (human SP-A) in association with specific lipids. In addition, the occurrence of several neutral and acidic glycosphingolipids in human and rat extracellular surfactants and rat alveolar type II cells is described. Selected components among the neutral glycolipids are bound by radiolabeled human SP-A; these are identified by TLC-LSIMS as predominantly ceramide mono- and disaccharides (human surfactant) and ceramide tri- and tetrasaccharides (rat surfactant and type II cells). A recombinant carbohydrate recognition domain (CRD) of human SP-A inhibits the binding of human SP-A to galactosyl ceramide and to galactose- and mannose-bovine serum albumin, indicating that the CRD is directly involved in the binding of SP-A to these ligands. These results provide evidence for a novel type of binding specificity for proteins that have Ca(2+)-dependent CRDs and raise the possibility that glycosphingolipids are endogenous ligands for SP-A.