No evidence for the presence of beta-casomorphins in human plasma after ingestion of cows' milk or milk products.
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Animals↗
Biomedical subjects
A Henschen
Publications and source records attributed to A Henschen.
Effects of 1% chlorhexidine gel treatment on sympathetic adrenergic nerves in human buccal mucosa.
The effect of treatment with a commercially available 1% chlorhexidine gel (Hibitane Dental) in custom trays for 14 days (10 min daily) on the sympathetic adrenergic nerves in biopsies from human buccal mucosa was investigated. Chemical assay of endogenous noradrenaline concentration and immunocytochemical demonstration of tyrosine hydroxylase served as markers for structural and functional integrity of the adrenergic nerves. The mean concentration of noradrenaline in chlorhexidine treated tissues did not differ from the mean concentration in control. No morphologic or fluorescence intensity differences of the tyrosine hydroxylase immunoreactivity were found in the chlorhexidine treated tissues when compared to controls. These findings indicate that a relatively intense treatment with a commercially available 1% chlorhexidine gel does not affect the adrenergic innervation of human buccal mucosa.
Adrenergic Fibers↗
Enkephalin and capsaicin-resistant substance P-like immunoreactivities in intra-ocular grafts of different fetal spinal cord areas.
Fetal spinal cord tissue was grafted to the anterior chamber of the eye of adult rats in order to evaluate survival and distribution of substance P- and enkephalin-immunoreactive neurons. Capsaicin treatment was used to evaluate any possible contribution of host iris-derived substance P fibres to the innervation of the grafts and to check for capsaicin sensitivity of graft substance P-positive systems. Substance P- and enkephalin-immunoreactive nerve fibres were present in grafts of half-transverse segments of the spinal cord and were clearly co-distributed throughout the grafts. Areas with a high density of substance P- and enkephalin-positive fibres resembling substantia gelatinosa were seen. Grafts of the dorsal horn alone had a moderate to high density of substance P- and enkephalin-positive fibres, while ventral horn grafts contained a low amount of such fibres. Capsaicin eliminated the substance P innervation of the host iris and the dorsal root ganglion-derived substance P innervation of the host spinal cord, while sparing the intrinsic substance P innervation of both host spinal cord and spinal cord grafts. These experiments show that intra-ocular grafts of defined spinal cord areas express relatively organotypic amounts of substance P- and enkephalin-positive nerve fibres, and thus emphasize the importance of intrinsic genetic determinants for spinal cord development.
Animals↗
Isolation and amino-acid sequence analysis of human sperm protamines P1 and P2. Occurrence of two forms of protamine P2.
The two protamines of human sperm cell nuclei, P1 and P2, were isolated in pure form after extraction with 6M guanidine/5% mercaptoethanol and alkylation with vinyl pyridine by reversed-phase high-performance liquid chromatography. The amino-acid sequence of protamine P1 was determined by analysing the intact protein and the fragments obtained by cyanogen bromide cleavage. Out of the 50 amino-acid residues 24 are arginines and 6 are cysteines. The sequence of protamine P2 was determined by analysing the intact protein and the fragments resulting from cleavage with endoproteinase Lys-C and thermolysin. Protamine P2 was found to occur in two forms which only differ in their N-terminal regions. The form P2' is three amino-acid residues longer at the N-terminus than the form P2''. Out of the 57 amino-acid residues in the longer form 27 are arginines and 5 are cysteines. Human protamine P1 is highly homologous with the protamines isolated from bull, boar, ram and mouse sperm cells, but human protamine P2 shows a novel type of structure, although also here the dominant amino acids are arginine and cysteine.
Amino Acid Sequence↗
Human Ile-Ser-bradykinin, identical with rat T-kinin, is a major permeability factor in ovarian carcinoma ascites.
Ascites from patients with metastatic ovarian carcinoma contains high amounts of an activity that increases vascular permeability, as easily detected by a rat skin test. Ascites was fractionated by gel permeation and reversed-phase high-performance liquid chromatographies. The fractions were analysed for permeability-increasing activity. In this way a peptide was isolated and identified as Ile-Ser-bradykinin by sequence and amino-acid analyses. It is identical with T-kinin which has previously been detected as a product of an acute-phase protein, T-kininogen, in rats but never in human material. The so far identified human kininogens, i.e. high- and low-molecular mass kininogens, can only release Met-Lys-bradykinin or its degradations products, as Ile-Ser-bradykinin is not a part of their structure. However, the present results provide evidence that the permeability factor Ile-Ser-bradykinin under certain conditions can be produced in considerable amounts also by human tissues.
Animals↗
[An evaluation of the information contained in the amino acid sequences of fibrinogen].
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Amino Acid Sequence↗
Characterization of fibrinogen Milano I: amino acid exchange gamma 330 Asp----Val impairs fibrin polymerization.
An abnormal fibrinogen was found in two asymptomatic members (father and daughter) of the same family, originating from northern Italy. Routine coagulation studies revealed prolonged thrombin and reptilase clotting times. Plasma fibrinogen levels, as determined by a functional assay, were markedly diminished, whereas the heat precipitation method indicated normal fibrinogen values. On the basis of these findings, a tentative diagnosis of dysfibrinogenemia was made, and according to the accepted nomenclature, this fibrinogen variant was called "fibrinogen Milano l." The time course of fibrinopeptide A and B release from fibrinogen Milano l was normal, but the aggregation of fibrin monomers was delayed. Two-dimensional electrophoresis of reduced variant fibrinogen chains showed a defective gamma-chain with increased cathodic mobility. An abnormal electrophoretic mobility was observed also for the gamma-chain remnants of fibrinogen fragments D1 and D2 derived from fibrinogen Milano l, whereas the charge anomaly was lost after a further digestion by plasmin to D3, suggesting that the structure abnormality of this variant is situated in the region gamma 303-356. An abnormal peptide was isolated after cyanogen bromide cleavage of intact fibrinogen Milano l. This fragment spans from position gamma 311 to gamma 336. Amino acid analysis of the abnormal peptide showed the presence of valine and a diminished content of aspartic acid. Sequence analysis demonstrated an amino acid exchange Asp----Val in the gamma-chain at position 330.
Adult↗
On the evolution of mammalian kininogens.
A model for the evolution of mammalian kininogens has been constructed which is based on sequence homology. This model suggests that several gene elements coding for the major constituents of the kininogens, i.e. the heavy chain, the light chain and the kinin segment, arose independently from their progenitors. The heavy chain of the kininogens known to execute their inhibitory function against cysteine proteinases has evolved by gene triplication from an ancestor which is closely related to the present-day form of the low molecular weight cysteine proteinase inhibitor cystatin. Part of the light chain of the mammalian kininogens which contains a unique region extremely rich in histidine and glycine has descended by gene multiplication from smaller units, possibly even from the dipeptide Gly-His. At present, the mode and timepoint of fusion of the various gene elements coding for the various structural constituents of the mammalian kininogens is not known.
Amino Acid Sequence↗
The amino acid sequence of the light chain of human high-molecular-mass kininogen.
The complete amino acid sequence of the light chain of human high-molecular-mass kininogen has been determined. The peptide chain contains 255 amino acid residues. The half-cystine, which forms the disulfide bridge to the heavy chain, was identified in position 225. Nine carbohydrate attachment sites were found. All carbohydrate side chains are O-glycosidically linked. Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. The histidine-rich region may have arisen by gene multiplication during evolution.
Amino Acid Sequence↗
Primary structure of elongation factor 2 around the site of ADP-ribosylation is highly conserved from archaebacteria to eukaryotes.
Elongation factor 2 (EF-2) from eukaryotes and archaebacteria can be ADP-ribosylated by diphtheria toxin (DT) [(1977) Annu. Rev. Biochem. 46, 69-94; (1980) Nature 287, 250-251]. The primary structure of the ADP-ribose accepting region in EFs from the archaebacteria Thermoplasma acidophilum Halobacterium cutirubrum and Methanococcus vannielli was determined in order to elucidate the degree of conservation compared with 4 previously established eukaryotic sequences [(1971) FEBS Lett. 103, 253-255]. Within a 9-residue sequence including the site of ADP-ribosylation 5 positions were found to be occupied by the same amino acid in all the archaebacterial and eukaryotic factors studied. There were more differences among the 3 archaebacterial sequences than among the 4 eukaryotic ones.
Adenosine Diphosphate Ribose↗
Limited proteolysis of human low-molecular-mass kininogen by tissue kallikrein. Isolation and characterization of the heavy and the light chains.
The limited proteolysis of human low-molecular-mass kininogen by kallikrein from tissue sources has been studied. Porcine pancreatic kallikrein applied in catalytic amounts split the kininogen molecule (apparent mass 68 kDa) with the release of lysyl-bradykinin (1 kDa). This generated a nicked kininogen molecule with a heavy chain and light chain interconnected via disulfide bridging. Following reductive cleavage of the disulfide bonds, the heavy chain of apparent mass 62 kDa was isolated by preparative sodium dodecyl sulfate electrophoresis, and the light chain of 5 kDa by reversed-phase high-performance liquid chromatography. The light chain was found to be composed of 38 amino acids with a single half-cystine residue. Amino-terminal sequence analysis revealed that the light chain is derived from the carboxy terminus of the kininogen molecule [Lottspeich et al. (1984) Eur. J. Biochem. 142, 227-232]. Immunological characterization of the isolated L chain indicated that it harbours antigenic site(s) unique for low-Mr kininogen as well as sites common to high-Mr and low-Mr kininogen.
Amino Acids↗
Novel opioid peptides derived from mitochondrial cytochrome b: cytochrophins.
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Animals↗
Thrombin-induced fibrinopeptide release from a fibrinogen variant (fibrinogen Sydney I) with an Aalpha Arg-16----His substitution.
Fibrinogen, purified from a recently identified case of dysfibrinogenaemia, fibrinogen Sydney I, was shown by thrombin digestion, high-performance liquid chromatography (HPLC) and amino acid analysis to be a heterozygous case of an A alpha Arg-16----His substitution. Kinetic studies have been carried out on the thrombin-induced release of fibrinopeptide A (FPA), fibrinopeptide B (FPB) and the variant peptide [His16]FPA. When thrombin was added to fibrinogen Sydney I at a concentration of 0.2 U/ml release of FPA was rapid and there was a 79-fold reduced rate of release of [His16]FPA, but the rate of release of FPB was not appreciably reduced. In contrast, at lower thrombin concentrations the rate of FPB release was reduced in proportion to the rate of total FPA release, supporting the view that release of fibrinopeptides is a sequential process. The second-order kinetic constant kcat/Km for hydrolysis of the abnormal A alpha chain by thrombin was calculated from Lineweaver-Burk plots to be 16-30-fold less than that for the normal A alpha chain. Molecular modelling studies, using a refined model of the trypsin-pancreatic-trypsin-inhibitor complex have been used to suggest how the histidine at the P1 site can be accommodated within the enzyme hydrophobic active-site pocket.
Amino Acids↗
Amino acid sequence of the ;pathogenesis-related' leaf protein p14 from viroid-infected tomato reveals a new type of structurally unfamiliar proteins.
We have established the complete sequence of the 130 amino acid residues of the pathogenesis-related (PR) protein p14 accumulating in tomato leaves infected with the viroid of the spindle tuber disease of potato (PSTV) and partial sequences of the PR protein 1a which accumulates in tobacco mosaic virus-infected tobacco leaves. Both PR proteins are closely related to each other. However, no homology could be found between the sequence of p14 and any of the 3061 published protein sequences compiled in the protein sequence database at present. Thus, p14 represents not only the first completely sequenced PR plant protein but also a new type of structurally unfamiliar proteins whose biological function in the diseased plant remains to be elucidated.
Journal Article↗
Combined grafts of the ventral tegmental area and nucleus accumbens in oculo. Histochemical and electrophysiological characterization.
Dissection techniques and optimal donor stages have been established for constructing an isolated intraocular model of the ventral tegmental area (VTA)-accumbens system using intraocular sequential grafting. Single grafts including accumbens and VTA respectively survived and developed many organotypic features when taken from 15-17 day fetuses. Falck-Hillarp fluorescence histochemistry showed dopamine neurons and terminals in single VTA grafts, no or almost no catecholamine fibers in single accumbens grafts, and a well-developed VTA-accumbens dopamine pathway in combined grafts where cell bodies in the VTA part provided the accumbens part with a rich terminal network. A similar distribution was found using immunohistochemistry with antibodies directed against tyrosine hydroxylase. CCK-like immunoreactivity had a distribution that mimicked that of the catecholamine-containing system. Enkephalin-like immunoreactivity was found both in single VTA and in single accumbens pieces as well as in both parts of the double grafts. Cells with long-duration action potentials typical of dopamine neurons discharged at approximately 8 Hz in single VTA grafts and below 1 Hz in the VTA part of VTA-accumbens double grafts. Cells in the accumbens portion of double grafts had shorter action potential durations and fired at 10-20 Hz. Haloperidol increased discharge frequency in VTA neurons with long action potential durations while apomorphine reduced discharge markedly. Antidromic activation of putative dopamine neurons in the VTA part was obtained by electrical stimulation of the accumbens part. The indirect dopamine agonist + 3-methyl-phencyclidine slowed firing rates of neurons in the accumbens part of double grafts. Taken together, the histochemical and the electrophysiological data show that the intraocular VTA-accumbens system retains several of its normal structural and functional characteristics. It is proposed that the isolated VTA-accumbens projection can be used as a model to study the cellular mechanism of action of stimulant and opiate drugs of abuse.
Animals↗
Spinal cord grafts in oculo: survival, growth, histological organization and electrophysiological characteristics.
Fetal spinal cord tissue was grafted to the anterior chamber of the eye of adult recipients. Transverse segments from the cervical and high thoracic levels were divided in halves which were grafted directly or further divided into ventral horn and dorsal horn parts before grafting. Survival and intraocular growth was monitored through the cornea. Grafts from E14 to E16 grew to final sizes several times the initial size. The final size of E17 grafts was approximately similar to the initial size, while the final size of E18 and E19 grafts was considerably smaller than the size at grafting. All grafts were well vascularized from the host iris. Grafts from younger donors contained several neurons typical of spinal cord including alpha-moto-neuron-type cells. Cells were found in clusters in gray matter areas surrounded by white matter. Extracellular recording revealed many spontaneously active cells. Several had high sustained discharge (10-25 Hz) and large amplitudes. Many cells could be excited by stimulation of the graft surface via activation of local afferents. It is concluded that the capacity of fetal spinal cord tissue to survive grafting to the eye chamber is inversely related to the donor age. Before E17, large grafts retaining several morphological and electrophysiological characteristics of spinal cord are obtained. The intraocular spinal cord graft provides a useful model for studies of spinal cord development and, using co-grafting techniques, a model for spinal cord regeneration and functional connectivity.
Animals↗
Intraocular grafts of nucleus raphe dorsalis provide cografted spinal cord with a serotonergic innervation.
Nucleus raphe dorsalis was sequentially cografted with spinal cord to the anterior chamber of the eye of adult rats. After maturation in oculo, the double grafts were examined histologically and immunohistologically utilizing an antiserum to serotonin (5HT). No 5HT-like immunoreactivity could be detected in grafts of single spinal cord or in irides of nongrafted eyes. In single raphe grafts, positive cell bodies and a dense terminal plexus were found. 5HT-positive fibers also penetrated into the irides forming dense networks in the walls of blood vessels and elsewhere in the irides. When spinal cord was cografted to maturated nucleus raphe dorsalis grafts, the raphe graft provided the spinal cord graft with a serotonergic innervation. Interestingly, when the grafting procedure was done in reversed order, i.e., the spinal cord was grafted first and left to mature before immature raphe grafts were added to the eye, the average density of 5HT-positive nerve terminals in the spinal cord part was clearly higher. It is concluded that immature as well as mature isolated spinal cord tissue can become innervated by adjacent central 5HT-neurons. Thus isolated replicas of descending serotonergic pathways to the spinal cord, suitable for electrophysiological analysis, are obtained in oculo.
Animals↗
Acidic acrosin inhibitors from bull seminal plasma. Structural differences.
The three acidic acrosin inhibitors of bull seminal plasma, BUSI I A, BUSI I B1 and BUSI I B2 were compared by thin-layer chromatographic and high-performance liquid chromatographic fingerprint analyses of the tryptic digests prepared from their S-carboxymethylated derivatives. It was found that the inhibitors differ only in their N-terminal regions. The inhibitor BUSI I B1 has a blocked N-terminus due to a pyroglutamic-acid residue. This residue is substituted by glutamic acid in BUSI I B2. The third inhibitor, BUSI I A, is four residues shorter at the N-terminus than the two other inhibitors. A high-performance liquid chromatography-based method for the separation of the three inhibitor variants was developed.
Acrosin↗