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Biomedical subjects

W Rapp

Publications and source records attributed to W Rapp.

At least 19 recordsLinked to original sources
PubMed1998

A new hybrid resin for stepwise screening of peptide libraries combined with single bead Edman sequencing.

A library system was developed for the discovery of bioactive peptides. Library synthesis and peptide sequencing was performed on a solid support while the screening for bioactivity was done with peptides in solution. The peptides were synthesized by split and mix, one-bead-one-peptide library synthesis, using a Tentagel S-NH2 solid support with a loading of approximately 100 pmol/bead. The major part of the peptide was connected to the support by a single acid-labile linker and a minor part of the peptide was acid-stabile attached to the polymer. The percentage of acid-stabile attached peptides could easily be controlled during modification of the amino functionalities of the resin at the start of the process. The cleavage rate of the acid-labile attached peptide from the resin depends on the composition of the cleavage mixture. When cleavage conditions were carefully controlled, a three-step partial cleavage protocol allowed for convergent bioactivity screening on peptide libraries using only one type of acid-labile linker. The partial cleavage and convergent screening procedure was repeated three times, after which the bead containing the bioactive peptide was sequenced. As such a bead still contained acid-stabile attached peptide, the Edman sequencing was straightforward and repetitive yields were excellent because the immobilized peptide was not washed out.

CD4-Positive T-Lymphocytes↗
PubMed1998

Coloured peptides: synthesis, properties and use in preparation of peptide sub-library kits.

Several methods were developed for the solid-phase synthesis (SPPS) of coloured peptides and peptide libraries. At first a bifunctional red compound, 4-(4-(N-ethyl-N-(3-(tert-butyloxycarbonyl)aminopropyl)amino)phenylazo)be nzoic acid (Boc-EPAB), was coupled with chloromethyl resin to obtain a new solid support suitable for SPPS using Boc chemistry. Peptides synthesized on this coloured resin had the chromophore at their C-termini. N-terminally coloured peptides were synthesized on a traditional solid support, coupled with chromophoric carboxylic acid before cleavage. A model pentapeptide, Phe-Ala-Val-Leu-Gly, and its ten derivatives were synthesized and their properties studied. It was found that the presence of chromophores decreases the water solubility of peptides. However, insertion of solubilizing tags (penta-lysine sequences or polyoxyethyl chains) into the molecule of any coloured derivative resulted in enhancement of the solubility. The RP-HPLC hydrophobicity indexes (phi0) of the coloured peptides were also determined because phi0 values are closely related to their water solubility. A coloured pentapeptide library was synthesized using the portioning-mixing method. Each component of this library contained the red azo dye (EPAB) and the penta-lysine tag. Before the last coupling step the samples were not mixed. All of the 19 sub-libraries obtained after cleavage were readily soluble in water, giving intense red solutions. The effect of chromophore (EPAB) and/or penta-lysine solubilizing tag on the biological activity was also studied. Potencies of the bovine neurotensin 8-13 fragment and its different coloured and penta-lysine derivatives were compared in isolated longitudinal muscle strips of guinea pig ileum. It was shown that the hexapeptide with penta-lysine tag had almost the same activity as the 8-13 fragment itself. The activity of the EPAB-derivative was found to be rather low. However, the presence of the solubilizing tag in the coloured hexapeptide compensated the negative effect of the chromophore.

Aminobenzoates↗
PubMed1998

Changes in reflex excitability following isometric contraction in humans.

Enhancement of muscle stretch following isometric contraction has been thought to occur as a result of inhibitory reflex mechanisms. Experiments with electrical stimulation (H-reflex) have demonstrated maximal H-reflex suppression during force relaxation followed by gradual recovery over the following 20 s. There has been considerable speculation as to whether electrical and mechanical stimulation elicit similar response behaviour. The present study examined postisometric reflex modulation following both stimulation modalities. In ten subjects dorsiflexion stimuli varying in speed and amplitude were applied after 30% and 60% maximal voluntary contraction (MVC). Modulation of the mechanically and electrically evoked responses following isometric plantarflexion was investigated. Reflex responses following both stimulation modalities were depressed during the course of force relaxation. A rather fast recovery was observed in mechanical stimulation. Postisometric response modulation was neither altered by the amount of isometric plantarflexion, nor by the amplitude of the applied stretch stimulus. With increasing velocity of the applied dorsiflexion, however, the shape of the reflex modulation persisted, but the magnitude of the responses was significantly enhanced. In electrical stimulation, however, recovery was delayed. It is suggested that postisometric reflex modulation is due to presynaptic inhibition. Moreover, possible peripheral mechanisms resulting from alpha-gamma-coactivation may also affect the stretch receptor itself because of inherent stiffness properties. The latter possibility particularly would explain the differences between mechanical and electrical stimulus modalities. With respect to practical implications, the very fast recovery (< 400 ms) of the stretch responses to control values strongly contradicts the interpretation that after isometric precontraction, suppression of reflex activity might be used for more efficient stretching of the tendomuscle system.

Adult↗
PubMed1994

Lipopeptide-polyoxyethylene conjugates as mitogens and adjuvants.

Two lipopeptide analogues of the Escherichia coli lipoprotein rendered water-soluble by polyoxyethylene were tested for mitogenicity in vitro in murine and human B lymphocytes and for adjuvant activity in vivo in mice. These highly amphiphilic lipopeptides retained the biological activity other lipopeptides usually exerted which supports the hypothesis of specific interactions of lipopeptides with membranes of reactive cells. The activation of human B lymphocytes by these lipopeptides was much less pronounced compared to that of murine cells. However, given in combination with anti-CD40 antibodies plus interleukin-4, human B lymphocytes could synergistically be stimulated to proliferate. As an adjuvant, the polyoxyethylene linked lipopeptides were almost as potent as Freund's adjuvants and other basic lipopeptides. Being water-soluble, these novel analogues are easy to apply and they are suitable for field studies as adjuvants when sonication can not usually be provided.

Adjuvants, Immunologic↗
PubMed1993

Recovery of stretch reflex responses following mechanical stimulation.

The recovery behaviour of mechanically evoked stretch responses was investigated. Stimuli which promoted identical dorsiflexing movements around the ankle joint were applied to ten subjects in two positions, seated and upright. The experimental sets comprised single as well as double dorsiflexing displacements. In the latter the stimuli were elicited for durations of either 100, 200 or 400 ms. Stretch responses following the first displacements were related to the stretch velocity but not to the amplitude. The responses of the plantar flexors following the second mechanical dorsiflexion were reduced with respect to the delay time between the first and second displacement. In addition, the magnitudes of these responses depended on the functional task: the stretch responses recovered much faster in the standing position when the triceps surae muscle was only slightly activated, whereas in the relaxed sitting position the reflexes remained suppressed. Both reciprocal inhibition, as well as the time course of the reformation of intrafusal cross-bridge links, may help to explain the depression of the monosynaptic stretch reflex.

Adult↗
PubMed1993

Orthotic devices in functional treatment of ankle sprain. Stabilizing effects during real movements.

Various orthotic devices generally employed for therapy of ankle sprain were tested under functional conditions. The experimental setup comprised inversion stimuli of 20 and 30 degrees (13 subjects) while standing on a tiltplatform as well as running movements of 8 and 12 km.h-1 on a treadmill (12 subjects). Ankle joint displacements were registered together with the surface EMGs from mm. peroneus long., tibialis ant., gastrocnemius med. and vastus med. In an additional study the relation between achilles tendon angle and talar tilt was determined under static conditions. Ten subjects with chronical instability were examined both in uni- and bipedal stance with the foot inverted to 20, 30 and 40 degrees. None of the tested devices could reduce achilles tendon angle movements completely neither during a sudden inversion nor during running. However, there were significant differences among the tested devices in the dorsal-plantar and in the eversion-inversion plane. In bipedal stance talar tilts were observed only when the foot was inverted more than 30 degrees. In unipedal stance these tilts were clearly reduced or disappeared totally. It is concluded that for functional rehabilitation, loading of the ankle joint is desirable in order to increase joint stability. The central importance of the treatment by orthotic devices may be regarded in an optimal passive joint stabilization combined with a correct fitting in order to preserve proprioceptive neuromuscular function for an active muscular stabilization.

Achilles Tendon↗
PubMed1992

Behaviour of triceps surae muscle-tendon complex in different jump conditions.

The force-length relationship of the human muscle-tendon complex (MTC) of the triceps surae and the achilles tendon was investigated in various stretch load conditions. Six male subjects performed various vertical jumps with maximal effort: squat jumps (SJ), counter movement jumps (CMJ) and drop jumps (DJ) from a height of 24 cm, 40 cm and 56 cm. The force-length relationship was calculated from the signals of the components of the ground reaction forces and the kinematic data obtained from the high-speed film records. Surface electromyograms (EMG) of the soleus, gastrocnemius and tibialis anterior muscles were also recorded. The force-length diagrams showed individually high sensitivity to the imposed stretch load. In conditions with relatively low stretch load requirements there was a counter-clockwise direction observable, indicating that the energy absorbed during the eccentric, or lengthening phase was lower than the energy delivered during the concentric, or shortening phase. In high load conditions this relationship was reversed indicating a negative energy balance. The EMG-length diagrams of SJ and CMJ consisted of an initial isometric loading of the muscle, followed by a shortening phase with only slightly reduced EMG amplitudes. In DJ, however, the diagrams showed an initial lengthening of the MTC with fairly constant activation amplitudes. After 40 ms an isometric loading of the muscle, lasting for approximately 80 ms, was followed by a shortening phase. It was concluded that segmental stretch reflex activation represented the predominant activation process during the isometric loading phase, to meet the adequate stiffness properties of the MTC.

Achilles Tendon↗
PubMed1992

Acute and chronic effects of low dose almitrine bismesylate in the treatment of chronic bronchitis and emphysema.

OBJECTIVES: Study of the acute and chronic effects of low-dose almitrine therapy in stable hypoxaemic patients with chronic bronchitis and emphysema. METHODS: A low daily dose of 75 mg almitrine bismesylate was administered for six months in 23 patients with chronic bronchitis and emphysema. Nine patients (group 1) were placed on oral almitrine bismesylate 25 mg t.i.d. after they had received a single intravenous dose of 60 mg almitrine three months earlier. Fourteen additional patients, seven receiving almitrine (group 2) and seven placebo (group 3) were randomized for a 6 month double-blind evaluation of both acute and chronic effects of 75 mg almitrine on pulmonary gas exchange and on pulmonary haemodynamics. All patients were followed-up with regular measurements of blood gases, body plethysmography and with evaluation of peripheral nerve function. RESULTS: Acute effects of almitrine were a significant increase in arterial oxygen tension by 14 mmHg after intravenous (p < 0.001) and by 15 mmHg after oral administration (p < 0.001), amelioration of hypercapnia, a slight transient increase in mean pulmonary artery pressure from 26 +/- 7 to 29 +/- 6 mmHg (NS) and a decrease of shunt due to improvement in ventilation/perfusion mismatching. In contrast, no acute changes in blood gases and pulmonary pressures were seen in the placebo group. A combination of almitrine with oxygen (8-10 L/min) was most effective in amelioration of hypoxaemia and shunt. With chronic almitrine therapy, the improvements in gas exchange persisted without elevation of pulmonary artery pressure (26 +/- 8 mmHg), whereas a negative trend in change of blood gases and pulmonary artery pressure occurred in the placebo treated group (NS). No significant changes in external ventilation, other spirometric parameters or adverse effects concerning peripheral nerve function were seen after almitrine or placebo treatment. The elimination of almitrine was fitted to a three compartment model and the terminal half-life in the patient population was found to be 32 +/- 29 days after intravenous dosing. CONCLUSION: Acute and six-month almitrine bismesylate therapy at a low daily dose of 75 mg is found to be safe, even in severely compromised patients, with regard to pulmonary haemodynamics and peripheral nerve function. The agent is beneficial to pulmonary gas exchange, with reduction of hypercapnia, of intrapulmonary shunt and also with regard to sustained elevation of arterial oxygen tension. A combination with inhaled oxygen seems especially efficacious.

Aged↗
PubMed1992

Radiation therapy of para-aortic lymph nodes in gynaecologic cancers: techniques, results and complications.

From 1965 to 1986 the para-aortic lymph nodes of 141 patients with gynaecologic cancers were irradiated directly after initial treatment of the primary site was completed. Another 32 patients only received para-aortic radiation treatment when para-aortic metastases became clinically apparent some time later. Radiation therapy was performed using a biaxial-four-segmental-rotating-field-technique. Crude survival of the group with initial para-aortic radiation treatment is far better than of the second group. Only five cases of severe complications due to para-aortic treatment were reported. In order to analyze the reason for such a low number of complications, CT-treatment planning for a typical patient was performed for different methods as they have been described in the literature, also referring to treatment data such as success, total dose and dose-distribution, thus influencing the complication rate. Obviously, the biaxial-four-segmental-rotating-field-technique is a method allowing to deposit a certain dose into the target volume without considerably damaging tissues in the vicinity.

Adolescent↗
PubMed1991

Cleavage kinetics and anchor linked intermediates in solid phase peptide amide synthesis.

Kinetics and cleavage conditions of peptide amide synthesis were studied using the anchor molecules 5-(4'-aminomethyl-3',5'-dimethoxyphenoxy)valeric acid (4-ADPV-OH) and 5-(2'-aminomethyl-3'-5'-dimethoxyphenoxy) valeric acid (2-ADPV-OH). Unexpectedly the anchor amide alanyl-4-ADPV-NH2 was isolated and characterized as an intermediate during the cleavage with trifluoroacetic acid (TFA) of alanyl-4-ADPV-alanyl-aminomethyl-polystyrene to yield the alanine amide. As a matter of fact the NH--CH alpha bond of the alanyl spacer has to be cleaved to form this intermediate. Using TFA-dichloromethane (1:9) alanyl-4-ADPV-NH2 was obtained as a cleavage product in 50% yield within 60 min, whereas the isomeric alanyl-2-ADPV-NH2 was formed more slowly under these mild conditions. At high TFA concentration no difference between the 2- and 4-ADPV anchor was observed in the rate of formation of the free alanine amide. The presence of tryptophan amide in the cleavage mixture resulted in an anchor alkylated tryptophan amide, which remains stable in acidic solution but disappears rapidly in the presence of the resin. A low TFA/high TFA cleavage procedure is recommended for peptide amid synthesis applying the ADPV anchor.

Alkylation↗
PubMed1981

Gastric marker proteins. Purification and immunohistological demonstration of the chief cell esterase.

An immunogenic acid stable esterase of gastric origin was purified from normal gastric juice. Specific antibodies were obtained by solid immunoadsorbents and were used for indirect immunoenzyme histology on gastric resection specimens. This esterase proved to be a cell specific marker for gastric chief cells. Different patterns of chief cell alterations in gastritis and cancer were observed.

Esterases↗
PubMed1980

Ectopic synthesis of pregnancy specific beta 1-glycoprotein (SP1) and placental specific tissue proteins (PP5, PP10, PP11, PP12) in nontrophoblastic malignant tumours. Possible markers in oncology.

By using an enzyme-bridge immunoperoxidase (PAP) technique, localization of so-called pregnancy-specific beta 1-glycoprotein (SP1) and placental-specific tissue proteins (PP5, PP10, PP11, PP12) was investigated in 19 cases of breast cancer, 24 cases of testicular malignant tumours, 12 cases of gastric cancer and some cases of other malignant tumours. These five glycoproteins isolated from human term placentae and characterized by Bohn are localised mainly in the cytoplasm, or nucleus of syncytiotrophoblast and appear to be specific for the trophoblast. But to a certain percentage these proteins could also be detected in the cytoplasma of malignant cells: In breast cancer, SP1 was present in 52.6% of cases, PP5 in 63.2%, PP10 in 68.4%, PP11 in 55.6% and PP12 in 31.6%. In testicular malignant tumours the detection rates of these proteins varied from 20.8 to 75.0% and in gastric cancer from 41.7 to 66.7%. In total 50% of the tumours tested were SP1-positive, 58.3% PP5-positive, 55.6% PP10-positive, 38.0% PP11-positive and 31.9% PP12-positive. In addition, it was found that mononuclear histiocytes showed a strong cytoplasmic staining for PP10 and PP12 and that a few other non-cancerous cells (i.e. striated cells in gastric metaplasia, gastric polyps etc.) stained for PP5, PP10, PP11 and/or PP12. All control sections were negative in malignant cells as well as in other tissue cells. This study confirms the reports that some kinds of malignant tumours produce SP1 and indicates that the ectopic production of proteins normally produced by the trophoblast is a common finding in malignant tumours. It furthermore suggests that such proteins may be useful as markers in monitoring patients with malignant diseases. The possible mechanisms of ectopic production of these inappropriate proteins in malignant tumours are discussed.

Breast Neoplasms↗