Synthesis and perspectives of new adjuvants and carrier systems for potential application for synthetic vaccines.
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Adjuvants, Immunologic↗
Biomedical subjects
G Jung
Publications and source records attributed to G Jung.
[Rotavirus and malabsorption. Immunofluorescence microscopy studies of small intestine specimens].
Only 20% of the total of diarrheas in childhood are caused by bacteria . Compared to it, very often the causes for such diseases are viruses, especially rotavirus. The virus - localisation is known to be in enterocytes of the duodenal or jejunal mucosa. So it is probable that a destruction of villous structure linked with diminishing of enzyme activities-especially of lactase--can occur. 53 mucosa specimens were investigated from children by means of fluorescence microscopy. Cryo-cut-slices (8-10 mu) were fixed in acetone and marked with self-prepared rotavirus-hyperimmuneserum (against SA11-virus) and examined using the microscope Fluoval. The distribution of fluorescence along the villi or flat surface was semiquantitatively evaluated. All the investigated children - aged 1-15 years - had been suspected for malabsorption. In 23 of them rotavirus antigen was detected and partly a low activity of disaccharidases, especially of lactase, but also there were simultaneously 7 cases of confirmed coeliac disease in this group. So we conclude: In some patients the rotavirus infection is a complication of malabsorption syndromes; or in other cases it is an asymptomatic accidental finding in the small intestine mucosa.
Adolescent↗
[Quantitative method of measuring platelet ATP released in vitro in the presence of various aggregating agents].
A new method for quantitative study of the release of ATP during platelet aggregation is described. The measures have been done by bioluminescence with ADP, collagen, arachidonic acid as inducing agents in 20 normal subjects. Different characteristics of quality of the method have been evaluated to define it as a "routine" test. The results, expressed in moles (M) of ATP by 1 X 10(6) platelets, are different according to the various inducing agents. In association with the study of the platelet aggregation this method should allow to have a better understanding of the anomalies of platelet functions in different diseases, and, may be, to define states of platelet "activation" as done by the study of the release of beta-thromboglobulin and of platelet factor 4.
Adenosine Diphosphate↗
[Temporary malabsorption following persistent rotavirus infection of the intestinal tract in children].
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Adolescent↗
Solvent-dependent structural features of the membrane active peptide trichotoxin A40 as reflected in its dielectric dispersion.
The dielectric dispersion of trichotoxin A40 in solvents of diverse lipophilicity (i.e. n-octanol, dioxane and mixtures of these) has been measured between 100 kHz and 50 MHz. In pure octanol also the weight average molecular weight and the concentration dependence of the circular dichroism spectrum were determined. An analysis of the data leads to the conclusion that in octanol an appreciable amount of the solute exists as single monomeric particles which are about 30 A long and 12.5 A in diameter. There is a dipole moment parallel to the long axis. In addition the evidence points at the presence of small aggregates (mainly dimers and trimers) formed by some kind of head-to-tail association. Evidently dioxane added to octanol favors a different structure of the trichotoxin monomer. It is apparently involved in another aggregation process resulting in aggregates of large size but comparatively low dipole moments.
Anti-Bacterial Agents↗
Quantitative determination of O-(beta-hydroxyethyl)-rutosides in serum by high-performance liquid chromatography.
A procedure for the quantitative determination of O-hydroxyethylated rutosides by high-performance liquid chromatography is described, which can be used for the detection of these modified flavonoids in human serum. Serum samples are processed by the addition of acetone, which removes most of the proteins. After passing the supernatant through a microcolumn of Amberlite XAD-2 and washing with water, the hydroxyethylated rutosides are eluted with methanol. The eluate is concentrated in vacuo. The methanolic solution of the residue is chromatographed on RP-8 columns using UV and fluorescence detectors. The mono- to tetrahydroxyethylated constituents and their corresponding aglycones could be separated with a step gradient, starting with a solvent system of water-methanol-acetic acid (70:30:6) followed by a mixture of water-ethanol-acetic acid (70:30:6). Alternatively, the rutosides can be separated by a linear gradient of water-acetonitrile. An almost linear calibration curve and about 80% recovery are obtained. A detection limit of 1 mg/l is achieved. Pharmacokinetic studies in human volunteers are described.
Chromatography, High Pressure Liquid↗
Modifications of plasma enzyme activities after severe head injury; evaluation of prognosis using multivariate methods.
The aim of this study was to analyse the plasma variations of four enzymatic activities (lactate dehydrogenase, aldolase, creatine kinase, aspartate aminotransferase) in 134 patients suffering from severe head injury. Enzymatic activities were assayed daily for 3 days after the trauma. Means of the four enzymatic activities were significantly different according to their evolution (death or survival), except for creatine kinase, 48 h after the trauma. Multivariate analysis indicated that lactate dehydrogenase and aldolase levels were useful in order to discriminate between potential survivors and non-survivors. The value of multivariate analysis in head traumatology is discussed.
Analysis of Variance↗
Melittin and a chemically modified trichotoxin form alamethicin-type multi-state pores.
The bee venom constituent, melittin, is structurally and functionally related to alamethicin. By forming solvent-free planar bilayers of small area (approx. 100 microns 2) on the tip of fire-polished glass pipettes we could observe single melittin pores in these membranes. An increase in the applied voltage induced further non-integral conductance levels. This indicates that melittin forms multi-level pores similar to those formed by alamethicin. Trichotoxin A40, an antibiotic analogue of alamethicin, also induces a voltage-dependent bilayer conductivity, but no stable pore states are resolved. However, chemical modification of the C-terminal molecule part by introduction of a dansyl group leads to a steeper voltage-dependence and pore state stabilization. Comparing structure and activity of several natural and synthetic amphiphilic polypeptides, we conclude that a lipophilic, N-terminal alpha-helical part of adequate length (dipole moment) and a large enough hydrophilic, C-terminal region are sufficient prerequisites for voltage-dependent formation of multi-state pores.
Alamethicin↗
Organizational forms of actin in 13762 ascites mammary tumor cell microvilli.
The organization of microvillus actin and its associated proteins have been investigated in sublines of mammary ascites tumors (MAT) with mobile (MAT-B1) and immobile (MAT-C1) cell surface receptors. Microvilli isolated from these sublines differ in morphology (branched for MAT-C1 versus unbranched for MAT-B1) and the presence of a 58,000-dalton polypeptide (58K). 58K is found associated with MAT-C1 microvilli, microvillar cytoskeletons obtained by nonionic detergent extractions, and microvillar membranes prepared under conditions which depolymerize actin microfilaments. By extraction with actin-stabilizing buffers (isotonic Triton-Mg-ATP) microvillar actin can be fractionated into four forms. About 40% of the actin is sedimented at low speed (7,500g, 15 min). The pellets contain microfilaments; actin and alpha-actinin are the predominant proteins. High-speed pellets from these low-speed supernates contain about 10% of the actin as a transmembrane complex with a cell surface glycoprotein (cytoskeleton-associated glycoprotein, [CAG] 75-80,000 daltons) in MAT-B1 cells or with CAG and 58K in MAT-C1 cells. Transmembrane complexes can be purified from MAT-B1 and MAT-C1 microvillar membranes in Triton-containing buffer by gel filtration or sucrose density gradient centrifugation. The presence of only CAG and actin in the MAT-B1 transmembrane complex strongly suggests the direct interaction of actin and a cell surface component. The high-speed supernates contain soluble actin. By gel filtration or rate-zonal sucrose density gradient centrifugation about 30% of the microvillar actin is found as small oligomers and about 10% as G-actin in this extraction buffer. We suggest that the actin-containing transmembrane complexes may serve as membrane-association sites for oligomeric actin segments and microfilaments and as initiation sites for actin polymerization.
Actins↗
Identification of a cytoskeleton-associated glycoprotein from isolated microvilli of a mammary ascites tumor.
Microvilli isolated from MAT-C1 13762 ascites tumor cells were extracted with Triton X-100 in phosphate-buffered saline (PBS) to yield cytoskeletal residues. Analysis of the residues by two-dimensional isoelectric focusing-dodecyl sulfate electrophoresis and silver staining suggested that one of the major components is a glycoprotein (CAG). Neuraminidase treatments and glucosamine labeling demonstrated that CAG is a glycoprotein, and lactoperoxidase iodination showed its presence at the microvillar surface. DNase treatments and myosin affinity analysis suggested an association between CAG and the microvillar microfilaments. Thus, CAG has the properties expected of a transmembrane-linking molecule connecting the cell surface to the cytoskeleton.
Animals↗
Synthetic analogues of the N-terminal lipid part of bacterial lipoprotein are B-lymphocyte mitogens in vitro and in vivo.
Two analogues of the N-terminal part of bacterial lipoprotein, S-[2,3-bis(palmitoyloxy)-(2RS)propyl]-N-palmitoyl-(R)-cysteine methyl ester ("tripalmitoyl cysteine") and S [-2,3-bis(palmitoyloxy)-(2RS)-propyl]-N-palmitoyl-(R)-cysteinyl-(S)-seryl-(S)-seryl-(S)-asparaginyl-(S)-alanine ("tripalmitoyl pentapeptide") were synthesized and tested for mitogenic activity. The compounds were potent mitogens towards mouse spleen cell cultures, as measured by 3H-thymidine incorporation and by hemolytic plaque assays. This activity was not dependent on the presence of serum. Tripalmitoyl pentapeptide had little, if any, effect on thymocytes. When injected intravenously into Balb/c mice, the synthetic compounds induced splenomegaly and polyclonal B-cell activation, the latter was evident from an increase in the number of plaque-forming cells against trinitrophenylated sheep red blood cells. Thus, a synthetic fragment of a bacterial surface component was shown to exhibit marked biological activity in vitro as well as in vivo.
Animals↗
Isolation of actin-containing transmembrane complexes from ascites adenocarcinoma sublines having mobile and immobile receptors.
The molecular nature of the cell surface-cytoskeleton interaction in microvilli isolated from ascites 13762 rat mammary adenocarcinoma sublines with immobile (MAT-C1) and mobile (MAT-B1) receptors was investigated by extraction and fractionation studies on the microvillar membranes. Extraction of membranes from MAT-C1 cells with Triton X-100-containing buffers gave insoluble residues showing three major components by NaDodSO(4)/polyacrylamide gel electrophoresis: actin, a 58,000-dalton polypeptide, and a cell surface glycoprotein of 75,000-80,000 daltons. The ratio of these components in Triton X-100-insoluble residues, as determined by scintillation counting of bands from gels of [(3)H]leucine-labeled microvillar membranes, approached equimolar, suggesting a specific complex of the components. The three components of the putative complex cosedimented on sucrose density gradients of Triton X-100/buffer-treated membranes. Gel filtration on Sepharose 2B gave a peak included in the column that contained only the glycoprotein, actin, and 58,000-dalton polypeptide by one-dimensional NaDodSO(4) electrophoresis and by two-dimensional isoelectric focusing/NaDodSO(4) electrophoresis. The glycoprotein-actin association could be disrupted only under strongly denaturing conditions. Complex prepared from MAT-B1 microvillar membranes by Sepharose 2B gel filtration in Triton X-100-containing buffers contained actin and the glycoprotein but no 58,000-dalton polypeptide. From these results we propose that the cell surface-cytoskeleton interactions in the 13762 tumor cell microvilli involve direct association of actin with the cell surface glycoprotein. We further suggest that the 58,000-dalton polypeptide stabilizes the association of this complex with the microfilaments in the MAT-C1 microvilli, thereby stabilizing the microvilli and restricting cell surface receptor mobility.
Actins↗
Synthesis of the mitogenic S-[2,3-bis(palmitoyloxy)propyl]-N-palmitoylpentapeptide from Escherichia coli lipoprotein.
The N-terminal pentapeptide of the lipoprotein from the outer membrane of Escherichia coli was obtained by coupling S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-N-palmitoyl-(R)-cysteine to O-tert-butylseryl-O-tert-butyl-seryl-asparaginyl-alanine tert-butyl ester followed by deprotection with trifluoroacetic acid. The tetrapeptide was built up from alanine tert-butyl ester with N-9-fluorenylmethyloxycarbonyl protected amino acids. S-[2,3-Bis(palmitoyloxy)propyl]-N-palmitoylcysteine was obtained from N,N'-dipalmitoylcystine di-tert-butyl ester via reduction to the thiol, and S-alkylation with racemic 3-bromo-1,2-propanediol followed by esterification with palmitic acid in the presence of dicyclohexylcarbodiimide/dimethylaminopyridine and deprotection with trifluoroacetic acid. The compounds were characterized unequivocally by 13C-NMR and mass spectra. The diastereomers of S-[2,3-bis(palmitoyloxy)propyl]-N-palmitoylcysteine tert-butyl ester with opposite configuration at the propyl-C-2 atom could be separated on a silica-gel column.
Escherichia coli↗
An economical assay for HDL phosphatidyl choline.
An economical enzymatic assay for HDL phosphatidyl choline is described, as adapted for the Cobas-Bio analyser (Hoffmann LaRoche). This method entails the enzymatic cleavage of phosphatidyl choline by phospholipase C from B. cereus, hydrolysis of phosphoryl choline and enzymatic determination of choline with choline oxidase by an enzymatic colour test. This method provides consistent values and is, by comparison to the enzymatic UV method (assaying choline with choline kinase in an optical test procedure), simpler to perform, more precise, and less expensive.
Alcohol Oxidoreductases↗
Iturin AL--a new long chain iturin a possessing an unusual high content of C16-beta-amino acids.
From a strain of Bacillus subtilis a new antifungal peptidolipid complex of the iturin group was isolated. This antibiotic complex contained six lipophilic beta-amino acids with 3-amino-14-methylpentanoic acid as the predominant component. Iturin AL contains: 2 D-Asp, 1 L-Asp, 1 L-Glu, 1 L-Pro, 1 L-Ser, 1 D-Tyr and a mixture of 2.9% iso-C14-beta-amino acid, 30.7% n-C14-beta-amino acid, 15% iso-C15-beta-amino acid, 9% anteiso-C15-beta-amino acid, 35.3% iso-C-16-beta-amino acid and 4.5% n-C16-beta-amino acid. The structures of the beta-amino acids were determined by combined GLC/MS. FAB mass spectroscopy revealed three M+H+ peaks (1,043, 1,057, 1,071). Iturin AL could be resolved into six components by HPLC whose structures confirm the high amount of long chain beta-amino acids.
Amino Acids↗
[Diagnosis and treatment of chronic inflammatory intestinal diseases. 2. Crohn disease].
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Child↗
[Plasma variations in severe head injuries: prognosis and post-traumatic surveillance. I. Enzymes].
Variations of four enzymatic activities (aldolase, aspartate aminotransferase, creatine kinase and lactate dehydrogenase) were followed in human plasma for four days after head injury. Univariate analysis showed that each plasma enzyme activity significantly differed as early as 72 h after head injury according to the clinical evolution (survival or death). Multivariate analysis performed with these four tests allowed us to correctly divide, in terms of survival or death, 75 to 91% of unselected patients (n = 280). Combining these four tests increased discriminant power in severe head injury. Most of the patients who were misclassified according to their biochemical data received phenobarbital for treatment. Valuable prognostic information may thus be obtained by daily determinations of four enzymatic activities.
Adolescent↗
Synthesis and biological activities of the tri-L-alanine derivative of isonicotinic acid hydrazide.
N-(tert-Butyloxycarbonyl) tri-L-alanine was coupled to the hydrazide function of isonicotinic acid hydrazide followed by cleavage of the amino protective group. The resulting dihydrochloride of the tri-L-alanine derivative of isonicotinic acid hydrazide was characterized by 13C-NMR. The minimal inhibitory concentration of isonicotinic acid hydrazide was not improved by the peptide derivative, and competition experiments with tri-L-alanine demonstrated that tri-L-alanyl-isonicotinic acid hydrazide did not use the peptide transport system. Isonicotinic acid hydrazide and its tri-peptide derivative possessed the same activity against pathogenic mycobacteria and did not antagonize each other. The relatively high stability of the tri-peptide derivative against peptidases of Mycobacterium fortuitum was discussed as being responsible for the significantly weaker activity against this atypical mycobacterium strain. With the exception of peptidases of hog intestinal mucose, the tri-L-alanyl derivative of isonicotinic acid hydrazide was stable to pronase and alpha-chymotrypsin when compared to other peptides.
Anti-Bacterial Agents↗