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Biomedical subjects

H Brückner

Publications and source records attributed to H Brückner.

At least 19 recordsLinked to original sources

Liquid chromatographic determination of D- and L-amino acids by derivatization with o-phthaldialdehyde and N-isobutyryl-L-cysteine. Applications with reference to the analysis of peptidic antibiotics, toxins, drugs and pharmaceutically used amino acids.

In order to evaluate and extend the applicability of an analytical method that enables the quantitative and simultaneous high-performance liquid chromatographic determination of D- and L-amino acids (DL-AAs) by automated precolumn derivatization with o-phthaldialdehyde together with the chiral thiol N-isobutyryl-L-cysteine [J. Chromatogr., 666 (1994) 259] selected natural and synthetic bioactive peptides, as well as pharmaceutically used formulations of AA, were investigated and the amounts of D- and L-AA determined by fluorescence detection. Peptides containing cys(e)ine were oxidized with performic acid prior to hydrolysis with 6 M HCl, and those containing Trp were hydrolyzed with 4 M methanesulfonic acid (24 h at 110 degrees C in both cases). Peptides analyzed were the peptide antibiotics bacitracin, gramicidins A and S, polymyxin B, metanicin C, the peptide toxin malformin A and the peptide drugs D-Arg-[Hyp3,Thi5,8,D-Phe7]-bradykinin, beta-casomorphin and alpha s1-exorphin. Further, the enantiomeric ratios of pharmaceutically used AA formulations containing racemic DL-Ser, DL-Asp and DL-Met were determined, and the AA drugs L-Asp and L-Trp were tested negatively for the presence of the respective D-enantiomers. In two aqueous formulations of L-AA used for parenteral nutrition, low amounts of D-AA (0.1-0.9% with respect to certain L-AA enantiomers and of totally 128 mg and 149 mg D-AAs per liter infusion solution) were determined.

Amino Acid Sequence

Role of endothelial cytoskeleton in high-permeability edema due to botulinum C2 toxin in perfused rabbit lungs.

The cytoskeleton of the endothelial cell has been suggested to regulate endothelial barrier function. We investigated the role of actin in the maintenance of pulmonary capillary integrity in perfused rabbit lungs. As a tool for selective perturbation of actin, we employed Clostridium botulinum C2 toxin, which is composed of a membrane translocation component (C2II) and a component (C2I) effecting ADP-ribosylation of nonmuscle G-actin. ADP-ribosylated actin no longer capable of polymerization but acts as a barbed end-capping protein, thereby effecting selective loss of the nonmuscle F-actin content. In buffer-perfused rabbit lungs, combined application of both toxin components (range 50 pg/ml-5 ng/ml C2I) resulted in a time- and dose-dependent increase in the capillary filtration coefficient (Kfc) with concomitant edema formation. Only 300:600 pg/ml C2I:II sufficed to induce a > 10-fold rise of Kfc values within 110 min. This severe lung permeability increase occurred in the absence of vasomotor responses and potassium release or lactate dehydrogenase release. Application of each single toxin component displayed markedly reduced efficacy. Similar to the C2 toxin effect, severe permeability increase without concomitant hemodynamic changes was evoked by cytochalasin D, known to possess F-actin-disrupting properties. Preloading of lung cells with phallacidin, which in opposition to C2 toxin decreases F-actin depolymerization, significantly reduced the C2 toxin-induced increase in vascular permeability. Electron microscopic examination of C2 toxin-poisoned lungs showed early, extensive endothelial cell attenuations, followed by disruptions of the endothelial layer and marked interstitial edema formation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Morphological alterations accompanying the effect of peptaibiotics, alpha-aminoisobutyric acid-rich secondary metabolites of filamentous fungi, on Culex pipiens larvae.

The effect of different representatives of the group of peptaibiotics, alpha-amino-isobutyric acid rich secondary metabolites of filamentous fungi, on Culex pipiens larvae was studied. Light and transmission electron microscopy techniques were used to localize the intracellular damage and to determine the target organells for the mode of action of peptaibols in mosquito larvae. Though different in insecticidal activity, all tested compounds induced the same type of tissue damage, which was characterized by heavy challenge of mitochondria followed by partial swelling, crystaeolysis and destruction of mitochondrial walls. It is concluded that the mode of action of peptaibols in mosquito larvae is mediated through the damage of mitochondria. The structure-mosquitocidal effect of these compounds, their potential mode of action and role in the natural fungal entomopathogenic process are briefly discussed.

Alamethicin

Ventilation-perfusion relationships in isolated blood-free perfused rabbit lungs.

The multiple inert gas elimination technique (MIGET) was applied to blood-free perfused isolated rabbit lungs. Commonly accepted criteria for reliability of the method were found to be fulfilled in this model. Ventilation-perfusion (VA/Q) distributions in isolated control lungs corresponded to those repeatedly detected under physiological conditions. In particular, a narrow unimodal dispersion of perfusate flow was observed: perfusion of low-VA/Q areas ranged below 1% and shunt flow approximately 2-3%; perfusion of high-VA/Q regions was not detected. Gas flow was characterized by narrow dispersion in the midrange-VA/Q areas. Application of a low level of PEEP (1 cmH2O) reduced shunt flow to less than 1%, and low-VA/Q areas were no longer noted. By using this PEEP-level, stable gas exchange conditions were maintained for greater than 5 h of extracorporeal perfusion. Graded embolization with small air bubbles caused a typical rightward shift (to higher VA/Q ratios) of mean ventilation, associated with the appearance of high-VA/Q regions and an increase in dead space ventilation. Mean perfusion was shifted leftward, and shunt flow was approximately doubled. Whole lung lavage with saline for washout of surfactant evoked a progressive manifold increase in shunt flow, accompanied by a moderate rise of perfusate flow to low-VA/Q areas. We conclude that the MIGET can be applied to isolated blood-free perfused rabbit lungs for assessment of gas exchange and that typical patterns of VA/Q mismatch are reproduced in this model.

Animals

Automated enantioseparation of amino acids by derivatization with o-phthaldialdehyde and n-acylated cysteines.

The enantioseparation of standard mixtures composed of protein DL-amino acids was performed by reversed-phase high-performance liquid chromatography of the corresponding diastereomeric isoindolyl derivatives, formed by automated precolumn derivatization with o-phthaldialdehyde (OPA) and a series of N-acyl-L-cysteines(Acyl-Cys). A photodiode-array detector, operating at 338 nm, was used for detection. In order to evaluate systematically the influence of the structures of the acyl group in the chiral thiol reagents, a series of novel N-acyl-L-cysteines was synthesized [acyl = n-butyryl, isobutyryl (i-But), pivaloyl, benzoyl) and the chromatographic behaviour of the diastereomers formed was compared with those of already known reagents, N-acetyl-L-cysteine and N-ter.-butyloxycarbonyl-L-cysteine. All Acyl-Cys derivatives of DL-amino acids were resolved. In particular, i-But-Cys gave the highest resolutions for most of the amino acid enantiomers in comparison with the other Acyl-Cys. Investigation of yoghurt using OPA-acetyl-Cys demonstrated the applicability of the method to a complex food matrix and the occurrence of D-Asp, D-Glu and D-Ala in this dairy product.

Amino Acids

Retention behaviour of paracelsin peptides on reversed-phase silicas with varying n-alkyl chain length and ligand density.

As part of further investigations on the characterization of the ligand-induced conformational stabilization of peptides, two series of n-alkyldimethylsilyl bonded silicas have been prepared. In series A the n-alkyl chain length, n, of the bonded phase was varied between 1 and 20 carbon atoms at a constant ligand density. In series B the ligand density, alpha exp, was gradually changed from 0 to 4.1 mumol/m2 on a C1, C4, C6, C8 and C18 bonded phase. The retention behaviour of four peptides of the paracelsin family were examined under isocratic conditions, using a ternary mobile phase of water-methanol-acetonitrile (22:39:39, v/v/v). Plots of k' versus n showed pronounced maxima between n = 2 and 4 carbon atoms, followed by a decrease by a factor of 3 at n = 5, whereas above 5 carbon atoms only a slight increase in k' was observed. The selectivity behaviour of the paracelsins A-D can be mainly rationalized by interaction of the amphiphathic polypeptide 3.6(13) (alpha)-helix with the hydrophobic ligand and protrusion of the key amino acid residues at positions 6 and 9 in the sequence. However, from experiments with a polystyrene stationary phase it is evident that hydrophobic interactions and different partition coefficients also contribute to the resolution of the paracelsin peptides. Furthermore, Van 't Hoff plots confirm significant free energy changes associated with retention. These observations provide the basis for evaluating the enthalpic and entropic changes associated with peptide interactions with n-alkyl silicas.

Amino Acid Sequence

[Development of the dental arch and the palate from a prosthodontic point of view].

The replacement of missing teeth by a fixed restoration in the juvenile dentition presents various problems. One of them is that the growth and development of the entire masticatory organ continues until around the 18th year of age. It was the goal of a longitudinal study to find out when a fixed tooth replacement can be used in an adolescent without impairing the final development of the jaw bones. For this reason the transverse and sagittal dimensions of the dental arches of male adolescents aged 14 to 18 years were measured every year, and the palatal surfaces were measured in the first and last year of the study. The values determined for the dental arches remained constant. The mean values and deviations revealed no significant differences. From a prosthodontic point of view the development of the dental arches and the palate may be considered as concluded.

Adolescent

[The performance of weaned piglets after a five week weaning period in relation to the weaning weight, the feed composition and the use of an effective ergotropic].

In a three-factorial experiment with 216 piglets of the same age (33 +/- 4 days) but a different weight (weaning weight from 5.5 to 9 kg) a conventional piglet rearing feed containing 20% crude protein (standard), a high-energy diet and a low-protein diet (16% crude protein) were tested. In three weaning weight classes the three diets were fed without or with Bisergon [2-N-(2-hydroxyethyl)-carbamoyl-3-methylquinoxaline-1.4-dioxide] over a period of 21 days. From the 22nd to the 71st day after weaning all the piglets received the standard diet without ergotropic. Among the factors studied the Bisergon supplement had the biggest effect. The additional weight gain varied from 9% in case of heavy piglets fed the high energy diet to 150% in case of light piglets fed the low protein diet. The ergotropic improved feed conversion by 22% and lowered the frequency of gastrointestinal diseases requiring treatment by 28%. At the end of the rearing period the weight of the piglets initially fed with Bisergon (means = 34.6 kg) was significantly higher than that of the control animals (means = 31.6 kg). The weaning weight and the diets influenced the final weight especially in the groups without ergotropic. In this case the piglets fed with the high-energy diet gained 30% more and those fed with the low-protein diet 40% less compared with the standard group. The average live weight gain of the light piglets (weaning weight 6.1 kg) was 30% lower than that of the heavier piglets (weaning weight 8.5 kg). The frequency of treatment against diarrhoea of the heavy piglets was by one third lower than that of the light animals. The piglets fed with the low-protein diet appeared to show a limitation of gastrointestinal diseases, but these animals also had the lowest feed intake. Independent of the Bisergon supplementation the weaning weight clearly influenced the live weight after the 10-week experiment. The initially better development of the animals started with the high-energy diet was compensated. In contrast to this, the piglets weighing 6.1 and 7.2 kg fed with the low-protein diet could not compensate the retarded development in comparison with the piglets fed with the standard or the high-energy diet. This experiment confirms the enormous improvement of the performance and of the health status of weaned piglets caused by the ergotropic Bisergon.(ABSTRACT TRUNCATED AT 400 WORDS)

Animal Feed

Determination of alpha-alkyl-alpha-amino acids and alpha-amino alcohols by chiral-phase capillary gas chromatography and reverse-phase high-performance liquid chromatography.

The enantiomeric resolution by fused-silica capillary gas-liquid chromatography (GLC) of non-protein DL-alpha-alkyl-alpha-amino acids of the structure H2NCR1R2COOH (R1 = alkyl, R2 = alkyl, alkaryl) was investigated by using chiral [L-valine-tert.-butylamide, linked to a statistical polymer of dimethylsiloxane and (2-carboxypropyl)methylsiloxane, Chirasil-L-Val, and XE-60-S-Val-S-alpha-phenylethylamide] and non-chiral (methylphenylcyanopropylvinylpolysiloxane, CP-Sil-19 stationary phases. To evaluate the resolution coefficients, N-acylamino acid n-propyl esters (acyl = acetyl, propionyl, trifluoroacetyl, pentafluoropropionyl, heptafluorobutyryl) and diastereomeric esters with S(-)-2-methyl-1-butanol, S(+)-2-butanol and S(+)-2-octanol were used. Although alpha-alkyl-alpha-amino acids in general gave lower resolution coefficients than the enantiomers of protein amino acids, most alpha-alkyl-alpha-amino acids could be resolved by using suitable derivatization procedures and, preferably, isothermal conditions. In addition, a number of DL-alpha-alkyl-alpha-amino acids could be separated by ligand-exchange chromatography (L-hydroxyproline/Cu2+) by both thin-layer chromatography (Chiralplate) and high-performance liquid chromatography (HPLC) (Nucleosil Chiral-1). Further, a standard mixture composed of fifteen alpha-amino acids and eleven alpha-amino alcohols could be completely separated by C18 HPLC after derivatization with o-phthaldialdehyde-2-mercaptoethanol (OPA-2-ME). The time and temperature dependences of the relative fluorescence of the adducts were investigated kinetically.

Amino Acids

Crystal structure of L-Pro-L-Leu-Aib-Aib-L-Glu-L-Valol, the C-terminal hexapeptide fragment of trichotoxin.

The crystal structure of L-prolyl-L-leucyl-alpha-aminoisobutyryl-alpha- aminoiso-butyryl-alpha-L-glutamyl-L-valinol (L-Pro-L-Leu-Aib-Aib-L-Glu-L-Valol), the C-terminal hexapeptide fragment of trichotoxin, has been determined by X-ray crystallography. The hexapeptide forms a right-handed 3(10)-helix consisting of two 10-atom hydrogen-bonded beta-turns of type III and one beta-turn of type I. Two of the intramolecular hydrogen-bonds are particularly weak, thus suggesting that the title compound may adopt non-helical conformations in solution, as observed by circular dichroism. In the crystal the molecules are hydrogen-bonded head-to-tail, forming infinitely long helical columns.

Amino Acid Sequence

Crystal structure of Z-Aib-Aib-Aib-Ala-Ala-Aib-OtBu, a hexapeptide fragment of trichotoxin.

The crystal structure of Z-Aib-Aib-Aib-Ala-Ala-Aib-OtBu, an end-protected hexapeptide with a sequence corresponding to residues 7-12 of several trichotoxin A-50 sequence analogues has been determined by X-ray crystallography. The hexapeptide adopts a right-handed 3(10)-helical conformation consisting of four consecutive beta-turns of type III. The helix is stabilized by four intramolecular hydrogen bonds. In the crystal the molecules are connected head to tail with intermolecular hydrogen bonding interactions among translationally related molecules thus forming infinitely long helical columns. The column-column interactions in the crystal are hydrophobic and occur predominantly between antiparallel directed columns.

Anti-Bacterial Agents

[Neurogenic intermittent claudication--occurrence and causes].

The topic dealt with is about frequency and causes of the neural claudication intermittens among patients of our neurological department as a specific clinical form of lumbosacral disorder syndromes. Concluding special attention is given to diagnostical and therapeutical problems.

Aged

Trichotoxin A40. Purification by counter-current distribution and sequencing of isolated fragments.

The isolation of the membrane-modifying polypeptide antibiotics from the mycelium of Trichoderma viride 5242 was optimized via extraction with dichloromethane and chromatography on Sephadex LH-20. The components trichotoxin A40 and A50 were separated from each other and purified by multiplicative counter-current distribution. The sequence of proteinase-resistant trichotoxin A40 was determined by combined gas chromatography and mass spectrometry of three isolated N-acetylated dodecapeptides and two N-prolylhexapeptides obtained after selective trifluoroacetolysis. Including amino acid exchanges due to natural microheterogeneity, the sequence is Ac-Aib-Gly(LAla)-Aib-LLeu-Aib-LGln-Aib-Aib-Aib(LAla )-LAla-Aib-Aib-LPro-LLeu -Aib-DIva(Aib)-LGlu-LValol. In contrast to the eicosapeptide alamethicin, trichotoxin A40 contains only 18 residues, with a higher proportion of alpha-aminoisobutyric acid (Aib), C-terminal L-valinol (Vol), one D-isovaline (Iva) and no proline at the N-terminal part.

Amino Acid Sequence

[Breast reconstruction following radical mastectomy with myocutaneous latissimus dorsi flap].

Reconstruction of the female breast following radical mastectomy is bond to some criteria of cancer. Various methods are possible. At present the best method is the amplification of the skin cover by a latissimus-dorsi-myocutaneous flap in combination with augmentation by prosthesis. The other breast is adapted to size and level of the reconstructed breast. A quarter to half a year later the latter will be completed by substitution of the areola. This not too difficult surgical procedure yields good results and relieves the women from the psychical burden of being mutilated.

Breast