PubMed Health⌕ Search

Biomedical subjects

J Kopp

Publications and source records attributed to J Kopp.

At least 55 records · Page 3Linked to original sources

Structure and mechanism of the amphibolic enzyme D-ribulose-5-phosphate 3-epimerase from potato chloroplasts.

Ribulose-5-phosphate 3-epimerase (EC 5.1.3.1) catalyzes the interconversion of ribulose-5-phosphate and xylulose-5-phosphate in the Calvin cycle and in the oxidative pentose phosphate pathway. The enzyme from potato chloroplasts was expressed in Escherichia coli, isolated and crystallized. The crystal structure was elucidated by multiple isomorphous replacement and refined at 2.3 A resolution. The enzyme is a homohexamer with D3 symmetry. The subunit chain fold is a (beta alpha)8-barrel. A sequence comparison with homologous epimerases outlined the active center and indicated that all members of this family are likely to share the same catalytic mechanism. The substrate could be modeled by putting its phosphate onto the observed sulfate position and its epimerized C3 atom between two carboxylates that participate in an extensive hydrogen bonding system. A mutation confirmed the crucial role of one of these carboxylates. The geometry together with the conservation pattern suggests that the negative charge of the putative cis-ene-diolate intermediate is stabilized by the transient induced dipoles of a methionine sulfur "cushion", which is proton-free and therefore prevents isomerization instead of epimerization.

Amino Acid Sequence↗

HZE dosimetry in space using plastic track detectors.

Plastic nuclear track detectors were used to measure the contribution of High charge Z and energy E (HZE) particles to the radiation exposure of manned space missions. Results from numerous space missions in the orbit planned for the International Space Station are compared. The measurements cover the declining phase of the last solar cycle during the past 7 years and various shielding conditions inside the US Space Shuttle and the Russian MIR-station.

Collodion↗

Galanin and NPY, two peptides with multiple putative roles in the nervous system.

In the present brief overview we summarize results from several studies focusing on two neuropeptides, galanin and neuropeptide Y (NPY) in discrete neuronal systems, where they coexist with classic transmitters. On the basis of studies in different animal models we propose that these peptides may be involved in regulation of certain CNS functions and that drugs acting on their receptors may be of use in new therapeutic strategies. At the spinal level galanin and NPY are regulated in DRG neurons by nerve injury and in dorsal horn neurons by inflammation. It is possible that this leads to attenuation of pain sensitivity. Moreover, both peptides may exert trophic effects, for example to enhance regeneration. In the hypothalamic arcuate nucleus NPY and its receptors are part of the feeding circuitry, and we suggest that derangement of these NPY neurons may at least in part underlay the lethal phenotype of anorectic mice, which die 22 days postnatally after showing decreased food intake and growth retardation. Expression of NPY and NPY receptors is changed in the hippocampus of mice comparatively early after prion inoculation, indicating that this peptide system is affected in this spongiform degenerative disease in a region of importance for learning and memory. Finally, galanin is co-localized with classic monoamine transmitters in two central systems, the dorsal raphe serotonin neurons and the locus coeruleus noradrenergic neurons. In both cases galanin causes hyperpolarization (at high concentrations) and prolongs monoamine-induced outward currents (at low concentrations), thus modulating activity in two systems of importance for many brain functions including mood regulation. It may therefore be interesting to analyse to what extent drugs affecting galaninergic transmission also may be efficient in the treatment of, for example, depression.

Animals↗

Active dosimetry on recent space flights.

The radiation exposure inside the spacecraft in low earth orbit was investigated with a telescope based on two silicon planar detectors during three NASA shuttle-to-MIR missions (inclination 51.6 deg, altitude about 380 km). Count and dose rate profiles were measured, as well as separate linear energy transfer (LET) spectra, for the galactic cosmic rays (GCR) and the trapped radiation encountered in the South Atlantic Anomaly (SAA). Effective quality factors are deduced from the converted LET spectra (in water) in the range 0.1-120 keV micrometer-1 according to ICRP 60. Measured mission averaged dose rates in silicon are in the range 98-108 microGy d-1 and 137-178 microGy d-1 for the GCR and SAA contributions, respectively. The deduced effective quality factors are 2.95-3.29 (GCR) and 1.18-1.25 (SAA), resulting in mission averaged dose equivalent rates of 631-716 microSv d-1 for the comparable three missions.

Atlantic Ocean↗

[The study of new bio-membrane-bred epidermal cell grafts].

OBJECTIVE: The human epidermal cells were bred on a kind of bio-membrane, the bio-brane, in engineering a kind of new epidermal substitute, the bio-membrane bred cell graft. METHODS: Fresh and frozen grafts of biomembrane bred epidermal cells were transplanted into the full-thickness wounds of nude mice and those received simple Bio-brane were served as control. The wounds of the two groups were observed daily and biopsy was taken on the 3, 5, 7, 10, 21 and 35 days respectively. RESULTS: Epidermal cells could be cultured in vitro on the bio-membrane reaching the sub-saturated state of 60 to 70 percents. The bio-membrane after being grafted the epidermal cells continued to proliferate and differentiate to form a layer of new epidermis. There was no difference between the fresh and the frozen bio-membranes. CONCLUSION: Bio-membrane bred with epidermal cells could be a kind of ideal epidermal substitute.

Animals↗

Immunotherapy of B-cell lymphoma with CD3x19 bispecific antibodies: costimulation via CD28 prevents "veto" apoptosis of antibody-targeted cytotoxic T cells.

Bispecific antibodies (CD3x19) against the CD3epsilon-chain of the T-cell-receptor/CD3 complex and the CD19 antigen on B cells can target polyclonal, nontumor-specific T cells to B lymphoma cells. This induces T-cell activation, and generation of cytotoxic T cells (CTLs). These polyclonal CTLs, targeted by the CD3x19 bispecific antibodies, can lyse CD19(+) B-lymphoma cells. In a xenotransplant model in severe combined immunodeficiency deficient (SCID) mice, we and others observed that CD28 triggering is required for efficient elimination of B-lymphoma cells and cure from the tumor in addition to CD3x19 administration. We also showed that the activation and targeting of CTLs to the target cell by signal one alone, ie, the CD3x19 mab, induces T-cell death by apoptosis. In blocking experiments we showed that this "veto" apoptosis is mediated by the CD95/Fas ligand. Addition of anti-CD28 (signal 2) renders the T cells resistant for veto apoptosis both in vitro and in vivo. We therefore conclude that the role of costimulation in immunotherapy with bispecific antibodies or other T-cell-based immune strategies is not only to facilitate T-cell activation but also to prevent T-cell deletion by apoptosis.

Animals↗

Neuropeptide Y: some viewpoints on a multifaceted peptide in the normal and diseased nervous system.

Using immunohistochemical and in situ hybridization methodologies the localization of neuropeptide tyrosine (NPY) and two of its receptors, the Y1- and the Y2-receptor (R), has been analysed in various tissues in normal animals and animals subjected to different experimental procedures as well as animals with a genetic and an acquired disease. (1) Dorsal root ganglion (DRG) neurons are discussed with special focus on the effect of peripheral nerve injury. In normal DRG neurons NPY cannot be detected, whereas Y1-R mRNA and Y1-R-like immunoreactivity (LI) are strongly expressed. The Y1-Rs decorate the membrane of the cell soma and are not transported peripherally into the axonal branches. Y2-R mRNA levels are low. After axotomy there is a marked increase in NPY, a decrease in Y1-Rs and an increase in Y2-Rs. The Y2-R is transported centrifugally. These findings suggest that NPY-ergic mechanisms participate in the adaptive changes of sensory neurons in response to injury. (2) Using specific antibodies the cellular and subcellular localization of the Y1-R protein have been analysed in cerebral blood vessels. The results demonstrate high concentrations of receptors in smooth muscle cells around pial arterioles with lower numbers in large vessels on the basal surface of the brain. In many regions the receptors 'disappear' after the arterioles have entered the brain tissue. At the ultrastructural level the receptors are found both on the endothelial and peripheral side of the muscle cells as well as laterally, where muscle cells oppose each other. The receptor protein is often associated with small vesicles. No NPY-positive nerve fibers were found around the Y1-R-rich arterioles, but they were only seen around the arteries with low Y1-R levels. The Y1-R-rich arterioles were, however, seen close to numerous NPY-positive fibers originating from central interneurons. These findings raise the possibility that centrally originating NPY can influence cerebral blood flow, possibly by stimulating NPY-Rs on the peripheral side of the muscle cells. However, also blood borne NPY, released under special conditions, such as stress from sympathetic nerves and the adrenal medulla and transported with blood, may stimulate receptors on the endothelial side of the smooth muscle cells. (3) In the arcuate nucleus Y1- and Y2-Rs are found, whereby the Y1-Rs are located in its ventro-medial portion and co-localized with POMC peptides, and the Y2-R in its ventromedial part, partly co-localized with NPY. NPY nerve endings makes synaptic contact with the POMC/Y1-R-positive neurons. In a mouse model for genetic anorexia very high levels of NPY were observed in arcuate neurons as compared to control mice. However, NPY mRNA levels were not different between the two groups. Taken together these findings are in good agreement with the view that NPY in the arcuate nucleus plays an important role in regulating feeding behaviour. (4) After intracerebral prion inoculation in mice an upregulation of NPY mRNA levels was observed in CA3 pyramidal neurons, and this effect was seen at a time point just before the first behavioural symptoms were manifested. At approximately the same time there was a dramatic decrease in Y2-R binding in strata oriens and radiatum of the CA1 region of the hippocampus, whereas in other regions no changes or much smaller changes were observed. Also, there was only a very slight decrease in Y2-R mRNA levels in CA3 neurons. It thus appears as if the prion disease prevents ligand binding to the Y2-R, perhaps by influencing traffic of receptor proteins, possibly at the level of cell membrane-associated caveolae, which have been implicated in the conversion of normal protein to scrapie protein. It is possible that these changes in NPY-ergic mechanisms may underlie some of the central symptoms associated with the prion disease. (ABSTRACT TRUNCATED)

Animals↗

Results of dosimetric measurements in space missions.

Detector packages consisting of plastic nuclear track detectors, nuclear emulsions, and theromoluminescence detectors were exposed at different locations inside the space laboratory Spacelab and at the astronauts' body and in different sections of the MIR space station. Total dose, particle fluence rate and linear energy transfer (LET) spectra of heavy ions, number of nuclear disintegrations and fast neutron fluence rates were determined of each exposure. The dose equivalent received by the Payload specialists (PSs) were calculated from the measurements, they range from 190 microSv d-1 to 770 microSv d-1. Finally, a preliminary investigation of results from a particle telescope of two silicon detectors, first used in the last BIORACK mission on STS 76, is reported.

Cosmic Radiation↗

Localization of neuropeptide Y Y1 receptors in cerebral blood vessels.

The localization of neuropeptide Y (NPY) Y1 receptor (R) -like immunoreactivity (LI) has been studied in cerebral arteries and arterioles of the rat by immunohistochemistry using fluorescence, confocal, and electron microscopy. High levels of Y1-R-LI were observed in smooth muscle cells (SMCs) in the small arterioles of the pial arterial network, especially on the basal surface of the brain, and low levels in the major basal cerebral arteries. The levels of Y1-R-LI varied strongly between adjacent SMCs. Y1-R-LI was associated with small endocytosis vesicles, mainly on the outer surface of the SMCs, but also on their endothelial side and often laterally at the interface between two SMCs. NPY-immunoreactive (Ir) nerve fibers could not be detected in association with the Y1-R-rich small arterioles but only around arteries with low Y1-R levels. A dense network of central NPY-Ir nerve fibers in the superficial layers of the brain was lying close to the strongly Y1-R-Ir small arterioles. The results indicate that NPY has a profound effect on small arterioles of the brain acting on Y1-Rs, both on the peripheral and luminal side of the SMCs. However, the source of the endogenous ligand, NPY, remains unclear. NPY released from central neurons may play a role, in addition to blood-borne NPY.

Animals↗

Neuropeptide Y1 receptors in the rat genital tract.

Using in situ hybridization and immunohistochemistry, the expression of type 1 neuropeptide Y (NPY) receptors (Y1-Rs) has been demonstrated in the rat genital tract. In the male Y1-R mRNA and Y1-R-like immunoreactivity (LI) were found in smooth muscles of predominantly arterioles and small arteries inside testis. Fibers showing NPY-LI could not be detected within testis but only in the tunica albuginea. These Y1-Rs are suggested to mediate vasoconstriction, possibly activated by NPY released from nerves in the tunica albuginea. In the female rat Y1-R mRNA, but not Y1-R-LI was found in vascular smooth muscles of arteries in the ovary and oviduct. In the oviduct Y1-R mRNA was also detected in the non-vascular smooth muscle layer. Fibers showing NPY-LI were found around blood vessels both in the ovary and oviduct. In the female genital tract also Y1-Rs may thus be involved in regulatory mechanisms mediating, for example, vasoconstriction.

Animals↗

Results of dosimetric measurements in space missions.

Detector packages consisting of thermoluminescence detectors (TLDs), nuclear emulsions and plastic nuclear track detectors were exposed in different locations inside spacecraft. The detector systems, which supplement each other in their registration characteristics, allow the recording of biologically relevant portions of the radiation field independently. Results are presented and compared with calculations. Dose equivalents for the astronauts have been calculated based on the measurements; they lie between 190 microSv d-1 and 860 microSv d-1.

Astronauts↗

Dosimetric mapping in BIORACK on IML2.

Seven detector packages consisting of plastic nuclear track detectors, nuclear emulsions and thermoluminescence dosimeters were exposed in different locations inside BIORACK during the IML2 mission. The detectors supplement each other in their registration characteristics and cover well the different contributions of the space radiations to the dose. In this report, results are given on total dose measurements, cosmic ray flux and neutron dose. Total doses differ by up to a factor of 1.5 and heavy ion fluxes by more than a factor of 6 in the different locations. The results are compared with those of previous missions. The mission equivalent dose for the astronauts was calculated from the measurements to be 3.8 mSv.

Cosmic Radiation↗

Dosimetry on the Spacelab missions IML1 and IML2, and D2 and on MIR.

Detector packages consisting of plastic nuclear track detectors, nuclear emulsions, and thermoluminescence detectors were exposed inside BIORACK during the Spacelab missions IML1 and IML2, in different sections of the MIR space station, and inside the Spacelab module at rack front panels or stowage lockers and in the Spacelab tunnel during D2. In addition, during D2, each Payload Specialist (PS) has worn three permanent detector packages; one at the neck; one at the waist; and one at the ankle. Total dose measurements, particle fluence rate and LET spectra, number of nuclear disintegrations and neutron dose from this exposure are given in this report. The results are compared to theoretical calculations and to previous missions results. The dose equivalent (total radiation exposure) received by the PSs were calculated from the measurements and range from 190 to 770 microSv d-1. Finally, a cursory investigation of results from a particle telescope from two silicon detectors, first used in the last BIORACK mission on STS76, is reported.

Atlantic Ocean↗

Hepatic expression of mature transforming growth factor beta 1 in transgenic mice results in multiple tissue lesions.

Aberrant expression of transforming growth factor beta 1 (TGF-beta 1) has been implicated in a number of disease processes, particularly those involving fibrotic and inflammatory lesions. To determine the in vivo effects of overexpression of TGF-beta 1 on the function and structure of hepatic as well as extrahepatic tissues, transgenic mice were generated containing a fusion gene (Alb/TGF-beta 1) consisting of modified porcine TGF-beta 1 cDNA under the control of the regulatory elements of the mouse albumin gene. Five transgenic lines were developed, all of which expressed the Alb/TGF-beta 1 transgene selectively in hepatocytes. The transgenic line 25 expressing the highest level of the transgene in the liver also had high (> 10-fold over control) plasma levels of TGF-beta 1. Hepatic fibrosis and apoptotic death of hepatocytes developed in all the transgenic lines but was more pronounced in line 25. The fibrotic process was characterized by deposition of collagen around individual hepatocytes and within the space of Disse in a radiating linear pattern. Several extrahepatic lesions developed in line 25, including glomerulonephritis and renal failure, arteritis and myocarditis, as well as atrophic changes in pancreas and testis. The results from this transgenic model strongly support the proposed etiological role for TGF-beta 1 in a variety of fibrotic and inflammatory disorders. The transgenic model may also provide an appropriate paradigm for testing therapeutic interventions aimed at neutralizing the detrimental effects of this important cytokine.

Animals↗

Localization of neuropeptide Y Y1 receptors in the rat nervous system with special reference to somatic receptors on small dorsal root ganglion neurons.

Immunohistochemical staining with an antiserum against the neuropeptide Y (NPY)-Y1 receptor (Y1-R) protein was shown in rat small dorsal root ganglion (DRG) neurons, which also were Y1-R mRNA-positive and calcitonin gene-related peptide (CGRP)-positive. The Y1-R-like immunoreactivity was almost exclusively located in the somatic plasmalemma and in the perinuclear region. Intracellular recording showed that the Y1 agonist [Leu31,Pro34]NPY evoked an outward current in small DRG neurons, suggesting a functional somatic Y1-R. No evidence for axonal transport of Y1-R protein was obtained after analysis of the dorsal horn for double staining with CGRP, after dorsal rhizotomy, or after compression of dorsal roots and the sciatic nerve. It is proposed that blood-borne NPY released from sympathetic nerves and adrenal medulla is the endogenous ligand for the Y1 receptors on the small DRG neurons. Y1-R-positive neurons were also seen in the dorsal horn of spinal cord, the hypothalamic arcuate nucleus, pyramidal cells in the cerebral cortex, Purkinje and basket cells in the cerebellar cortex, and in many other brain regions.

Animals↗

Cologne Burn Centre experiences with glycerol-preserved allogeneic skin: Part I: Clinical experiences and histological findings (overgraft and sandwich technique).

In an effort to improve the take of finely meshed autografts a modification of the sandwich technique, as first published by Alexander et al. (1981), was developed. In contrast to the techniques described by other authors, the wound bed is sealed with fibrin glue spray after excision of the burns. Widely meshed autografts are then covered with non-meshed (only scarcely sliced) glycerolized allograft sheets, being fixed with staples. Patients are placed on fluidized beds and are exposed without dressings from the fifth day onwards. Histologically and clinically, it can be assumed that part of the glycerolized allodermis is incorporated. During the weeks after transplantation, a creeping substitution of the allodermis by autologous tissue takes place. This would suggest a co-existence between glycerol-preserved hypo-allergenic allografts and auto-epidermis. Research on the definitive fate of allodermis in cases of sandwich grafting is continuing.

Adult↗

Cultured autologous keratinocytes in fibrin glue suspension, exclusively and combined with STS-allograft (preliminary clinical and histological report of a new technique).

The use of cultured epidermal cell sheets has become a recognized method for the coverage of extensive burns. The disadvantages are a long time-lag until the cells are available, the fragility and difficult handling of the grafts, an unpredictable 'take' and extremely high costs. In three patients with deep partial and full skin thickness burns we have applied cultured autologous keratinocytes suspended in fibrin glue. In two of these patients the keratinocyte culture in the fibrin matrix (KFGS) was overgrafted with allogeneic, glycerine-preserved split thickness cadaver skin. The area thus covered ranged from 3 to 15 per cent TBSA. Cultured grafts were available between 2.5 and 3 weeks. The non-confluent cells developed a continuous epithelial layer within the 4 days until the first dressing change. Histological examination showed a stratified neoepidermis. Clinically the new skin had satisfactory stability and mechanical quality. The epidermis of the allogeneic overgrafts desquamated within a few days without signs of inflammation, but there are indications that the STS-allograft dermis is at least partly integrated into the new skin and may serve as a scaffold for the grafted cell culture. The fibrin glue matrix seems to give sufficient adherence stability to keratinocytes that are grafted in an actively proliferating state. Further advantages are the easy repetition and application, as well as a reduction in operating time and costs in these severely injured patients.

Adult↗