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Biomedical subjects

K Werner

Publications and source records attributed to K Werner.

At least 19 recordsLinked to original sources

In vitro fixator rod loading after transforaminal compared to anterior lumbar interbody fusion.

BACKGROUND: Cages are commonly used to assist lumbar interbody fusion. They are implanted from various approaches. In many cases internal fixators are added to provide sufficient stability. However, how the rods of these fixators are loaded and whether the kind of approach affects these loads is still unknown. The aim of this in vitro study therefore was to determine the loads acting on fixator rods and cages after anterior compared to transforaminal lumbar interbody fusion. METHODS: Six intact human lumbar spine specimens (L1-5) were loaded in a spine tester with pure moments (+/-7.5 N m) in the frontal, sagittal and transverse plane. Loading was repeated, first, after the segments L2-3 and L4-5 were instrumented either with an anterior or a transforaminal lumbar interbody fusion cage "stand alone" and, second, after additional stabilisation with an internal fixator. The rods of the fixator and the four "corners" of the cages were instrumented with strain gauges. FINDINGS: The loads transmitted through the rods were highest in lateral bending. In this loading direction an axial distraction force of in median up to 140 N, an axial compression force of up to 100 N, and a resultant bending moment of up to 1.1 N m were measured in each rod. These loads tended to be lower for the anterior compared to the transforaminal approach. For comparison, the load applied was +/-7.5 N m. The axial strains recorded in the four "corners" of the cages considerably varied from one specimen to the other. Differences in cage strain between the two approaches could not be detected. INTERPRETATION: The loads acting on the rods of the fixator were small compared to the load that was applied. Thus, other structures such as the cages or the facet joints still play an important role in load transfer. The type of approach (anterior or transforaminal) had only little effect on the loading of the rods. This also applies to the local loading of the cages, which probably more depends on the fit between cage and endplates and on the local stiffness properties of the adjacent vertebral bodies.

Aged↗

The autonomic and behavioral profile of emotional dysregulation.

The authors describe a patient with focal brain atrophy and emotional lability characterized by episodes of excessive crying and laughing. The patient was selectively impaired in the production of voluntary complex facial movements and was unable to regulate her emotional behavior and autonomic reactivity. She also displayed increased behavioral and autonomic changes when explicitly trying to suppress her responses to emotional stimuli (compared with when not trying to regulate her responses). This pattern of deficits supports a selective deficit in voluntary emotional control.

Atrophy↗

Novel mechanism of H0 dibaryon production in proton-proton interactions from parton-based Gribov-Regge theory.

A novel mechanism of H0 and strangelet production in hadronic interactions within the Gribov-Regge approach is presented. In this approach the H0 is produced by the same mechanism as usual hadrons, namely, by disintegration of the remnant formed by the exchange of pomerons between the two protons. Rapidity and transverse momentum spectra of the observed hadrons are well described in this approach. In contrast to traditional distillation approaches, here the production of multiple (strange) quark bags does not require large baryon densities or a quark gluon plasma. We calculate the rapidity and transverse momentum distributions as well as the 4pi multiplicity of the H0 for sqrt[s]=17 GeV (Super Proton Synchrotron) and 200 GeV (Relativistic Heavy Ion Collider). In both cases the H0, if it exists, should be observable by the present experiments.

Journal Article↗

rym15 from the Japanese cultivar Chikurin Ibaraki 1 is a new barley mild mosaic virus (BaMMV) resistance gene mapped on chromosome 6H.

Breeding for resistant cultivars is the only way to prevent high yield loss in barley caused by the soil-borne barley mild mosaic virus (BaMMV) complex. We have characterized the BaMMV resistance of barley cv. Chikurin Ibaraki 1. Doubled haploid lines were obtained from the F(1) between the susceptible six-rowed winter barley cultivar, Plaisant, and Chikurin Ibaraki 1. Each line was tested for reaction to BaMMV by mechanical inoculation followed by DAS-ELISA. Of 44 microsatellites that covered the genome, 22 polymorphic markers were tested on one susceptible and one resistant bulk, each comprising 30 lines. Differential markers and additional microsatellite markers in the same region were then tested on the whole population. A bootstrap analysis was used to compute confidence intervals of distances and to test the orders of the resistance gene and the closest markers. A segregation of 84 resistant/98 susceptible lines fitted a 1:1 ratio (chi(2)=1.08, P=0.30), which corresponds to a single gene in this DH lines population. The resistance gene was flanked by two markers near the centromeric region of chromosome 6HS-Bmag0173, at 0.6+/-1.2 cM, and EBmac0874, at 5.8 +/- 3.4 cM. We propose to name this new resistance gene rym15. This resistance gene and associated markers will increase the possibilities to breed efficiently for new cultivars resistant to the barley mosaic disease.

Chromosome Mapping↗

Dissection of resistance to soil-borne yellow-mosaic-inducing viruses of barley (BaMMV, BaYMV, BaYMV-2) in a complex breeders' cross by means of SSRs and simultaneous mapping of BaYMV/BaYMV-2 resistance of var. 'Chikurin Ibaraki 1'.

Ninety-three F(1)-derived doubled haploid (DH) lines from a complex breeders' cross involving the Japanese genotype 'Chikurin Ibaraki 1', which is resistant to barley mild mosaic virus (BaMMV) and two strains of barley yellow mosaic virus (BaYMV and BaYMV-2), three susceptible varieties ('Hamu', 'Julia' and a breeding line) and cv. 'Carola', which carries rym4 conferring resistance to BaMMV and BaYMV, were analysed for resistance to BaMMV, BaYMV and BaYMV-2. The DH lines fell into four phenotypic classes. In addition to completely resistant and susceptible genotypes, DHs were observed which were either resistant to BaMMV and BaYMV or to BaYMV and BaYMV-2. For BaMMV and BaYMV-2 resistance, segregation ratios approaching 1r:1s were observed, suggesting the presence of single resistance genes. In contrast, the segregation ratio for BaYMV fits a 3r:1s segregation ratio, suggesting the presence of two independently inherited genes. From the genetic analysis, we conclude that a resistance locus effective against BaYMV and BaYMV-2 originates from Chikurin Ibaraki 1 and segregates independently from the Carola-derived rym4 resistance that is effective against BaYMV and BaMMV. The BaMMV resistance in Chikurin Ibaraki 1 has probably been lost during population development. This hypothesis was tested using a simple-sequence repeat (SSR) marker (Bmac29) linked to rym4. All BaMMV-resistant DH lines supported amplification of the rym4-resistance diagnostic allele. To identify the genetic location of the Chikurin Ibaraki 1-derived resistance against BaYMV/BaYMV-2, bulked DNA samples were constructed from the four resistance classes, and bulked segregant analysis was performed using a genome-wide collection of SSRs. Differentiating alleles were observed at two linked SSRs on chromosome 5H. The location of this BaYMV/BaYMV-2 resistance locus was confirmed and further resolved by linkage analysis on the whole population using a total of five linked SSRs.

Genes, Plant↗

Overpopulation of Omega; in pp collisions: a way to distinguish statistical hadronization from string dynamics.

The Omega/Omega ratio originating from string decays is predicted to be larger than unity in proton-proton interactions at SPS energies ( E(lab) = 160 GeV). The antiomega dominance increases with decreasing beam energy. This surprising behavior is caused by the combinatorics of quark-antiquark production in small and low-mass strings. Since this behavior is not found in a statistical description of hadron production in proton-proton collisions, it may serve as a key observable to probe the hadronization mechanism in such collisions.

Journal Article↗

Effects of hypergravity on the development of cell number and asymmetry in fish brain nuclei.

Larval cichlid fish (Oreochromis mossambicus) siblings were subjected to 3 g hypergravity (hg) and total darkness for 21 days during development and subsequently processed for conventional histology. Further siblings reared at 1 g and alternating light/dark (12h:12h) conditions served as controls. Cell number counts of the visual Nucleus isthmi (Ni) versus the vestibular Nucleus magnocellularis (Nm) revealed that in experimental animals total cell number was decreased in the Ni, possibly due to retarded growth as a result of the lack of visual input whereas no effect was observed in the Nm. Calculating the percentual asymmetry in cell number (i.e., right vs. the left side of the brain), no effects of hg/darkness were seen in the Ni, whereas asymmetry was slightly increased in the Nm. Since the asymmetry of inner ear otoliths is decreased under hg, this finding may indicate efferent vestibular action of the CNS on the level of the Nm by means of a feedback mechanism.

Adaptation, Physiological↗

Characterization and identification of cytochrome P450 metabolites of arachidonic acid released by human peritoneal macrophages obtained from the pouch of Douglas.

Cytochrome P450 metabolism of arachidonic acid (AA) was investigated in human peritoneal macrophages which play a central role in chronic pelvic diseases in women (for example in endometriosis). The formation of eicosanoids other than prostaglandins (PGs) by these cells is still unknown. In non-activated macrophages obtained from women in the reproductive age, the main [(3)H]-AA metabolites coeluted with epoxyeicosatrienoic acids, dihydroxyeicosatrienoic acids (DHETs) and hydroxyeicosatetraenoic acids (HETEs) in reverse-phase HPLC. After zymosan activation a shift to PGs pathway was observed. Treatment with low doses of 2,3,7,8-tetrachlorodibenzo- p -dioxin increased the formation of a metabolite coeluting with 5,6-DHET. By gas chromatography/mass spectrometry 5,6-DHET (after beta-naphthoflavone induction), and 14,15-DHET as well as 11,12-DHET (after AA stimulation) were identified as major epoxygenase metabolites, respectively. The enantioselective formation of 12(S)-HETE was demonstrated by chiral-phase HPLC. Our findings demonstrate that non-activated peritoneal macrophages produce substantial amounts of bioactive cytochrome P450 metabolites of AA.

Arachidonic Acid↗

Self-consistency requirement in high-energy nuclear scattering.

Practically all serious calculations of exclusive particle production in ultrarelativistic nuclear or hadronic interactions are performed in the framework of Gribov-Regge theory or the eikonalized parton model scheme. It is the purpose of this paper to point out serious inconsistencies in the above-mentioned approaches. We demonstrate that requiring theoretical self-consistency reduces the freedom in modeling high-energy nuclear scattering enormously, and we introduce a fully self-consistent formulation of the multiple-scattering scheme in the framework of a Gribov-Regge--type effective theory. In addition, we develop new computational techniques which allow for the first time a satisfactory solution of the problem in the sense that calculations of observable quantities can be done strictly within a self-consistent formalism.

Journal Article↗

The apical membrane of intestinal brush cells possesses a specialised, but species-specific, composition of glycoconjugates--on-section and in vivo lectin labelling in rats, guinea-pigs and mice.

Brush cells are specialised epithelial cells that are assumed to represent chemoreceptors of the digestive tract. They comprise a small population of the epithelial cells lining the intestine, possess a unique ultrastructure and, in many aspects, resemble the receptor cells of taste buds. To characterise glycoconjugates possibly involved in a sensory function, we investigated brush cells in the small intestine of three species using lectin histochemistry in confocal light and thin-section electron microscopy. Brush cells of rats were selectively labelled by the sialic acid-specific lectin Maackia amurensis agglutinin, those of guinea-pigs by the D-galactose-specific lectin Bandeiraea simplicifolia agglutinin, isolectin B4 and those of mice by the L-fucose-specific lectin Ulex europaeus agglutinin lectin I. Lectin binding sites were consistently located in the glycocalyx of the apical membrane and in that of cytoplasmic vesicles. In vivo lectin labelling revealed that the glycoconjugates of the apical membrane are accessible under physiological conditions, that brush cells do not endocytose and that they probably possess a high membrane turnover rate. The results show that specialisations exist in the composition of glycoconjugates forming the glycocalyx of brush cells in all species investigated. The presence of brush cell-specific glycoconjugates would be in accordance with the current hypothesis of a receptive function of brush cells. Differences in the specific glycosylation patterns among rats, guinea-pigs and mice indicate that species-specific adaptations exist.

Animals↗

Fish otolith growth in 1g and 3g depends on the gravity vector.

Size and asymmetry (size difference between the left and the right side) as well as calcium (Ca) content of inner ear otoliths of larval cichlid fish Oreochromis mossambicus were determined after a long-term stay at hypergravity conditions (3g; centrifuge). Both utricular and saccular otoliths (lapilli and sagittae, respectively) were significantly smaller after hyper-g exposure as compared to parallely raised 1g-control specimens and the absolute amount of otolith-Ca was diminished. The asymmetry of sagittae was significantly increased in the experimental animals, whereas the respective asymmetry concerning lapilli was markedly decreased. In the course of another experiment larvae were raised in aquarium hatch baskets, from which one was placed directly above aeration equipment which resulted in random water circulation shifting the fish around ("shifted" specimens). The lapillar asymmetry of the "stationary" specimens showed a highly significant increase during early development when larvae were forced to lay on their sides due to their prominent yolk-sacs. In later developmental stages, when they began to swim freely, a dramatic decrease in lapillar asymmetry was apparent. Taken together with own previous findings according to which otolith growth stops after vestibular nerve transaction, the results presented here suggest that the growth and the development of bilateral asymmetry of otoliths is guided by the environmental gravity vector, obviously involving a feedback loop between the brain and the inner ear.

Animals↗

Compartmental pharmacokinetics and tissue distribution of multilamellar liposomal nystatin in rabbits.

The plasma pharmacokinetics of multilamellar liposomal nystatin were studied in normal, catheterized rabbits after single and multiple daily intravenous administration of dosages of 2, 4, and 6 mg/kg of body weight, and drug levels in tissues were assessed after multiple dosing. Concentrations of liposomal nystatin were measured as those of nystatin by a validated high-performance liquid chromatography method, and plasma concentration data were fitted into a two-compartment open model. Across the investigated dosage range, liposomal nystatin demonstrated nonlinear kinetics with more than proportional increases in the AUC(0-24) and decreasing clearance, consistent with dose-dependent tissue distribution and/or a dose-dependent elimination process. After single-dose administration, the mean C(max) increased from 13.07 microg/ml at 2 mg/kg to 41.91 microg/ml at 6 mg/kg (P < 0.001); the AUC(0-24) changed from 11.65 to 67.44 microg. h/ml (P < 0.001), the V(d) changed from 0.205 to 0. 184 liters/kg (not significant), the CL(t) from 0.173 to 0.101 liters/kg. h (P < 0.05), and terminal half-life from 0.96 to 1.51 h (P < 0.05). There were no significant changes in pharmacokinetic parameters after multiple dosing over 14 days. Assessment of tissue concentrations of nystatin near peak plasma levels after multiple dosing over 15 days revealed preferential distribution to the lungs, liver, and spleen at that time point. Substantial levels were also found in the urine, raising the possibility that renal excretion may play a significant role in drug elimination. Liposomal nystatin administered to rabbits was well tolerated and displayed nonlinear pharmacokinetics, potentially therapeutic peak plasma concentrations, and substantial penetration into tissues. Pharmacokinetic parameters were very similar to those observed in patients, thus validating results derived from infection models in the rabbit and allowing inferences to be made about the treatment of invasive fungal infections in humans.

Animals↗

High-performance liquid chromatographic determination of liposomal nystatin in plasma and tissues for pharmacokinetic and tissue distribution studies.

A reliable reversed-phase high-performance liquid chromatographic method was developed for the determination of liposomal nystatin in plasma. Nystatin is extracted by 1:2 (v/v) liquid-liquid extraction with methanol. Separation is achieved by HPLC after direct injection on a muBondapak C18 analytical column with a mobile phase composed of 10 mM sodium phosphate, 1 mM EDTA, 30% methanol and 30% acetonitrile adjusted to pH 6. Detection is by ultraviolet absorbance at 305 nm. Quantitation is based on the sum of the peak area concentration of the two major isomers of nystatin, which elute at 7.5-8.5 and 9.5-10.5 min. The assay was linear over the concentration range of 0.05 to 50 microg/ml. The lower limit of quantitation was 0.05 microg/ml, sufficient for investigating the plasma pharmacokinetics of liposomal nystatin in preclinical studies. Accuracies and intra- and inter-day precision showed good reproducibility. With minor modifications, this method also was used for assaying nystatin in various non-plasma body fluids and tissues.

Acetonitriles↗

Novel phenotypes and developmental arrest in early embryo specific mutants of maize

Embryo specific (emb) mutants exhibit aberrant embryo development without deleterious effects on endosperm development. We have analyzed five emb mutants of maize, which, based on their developmental profiles can be divided into two groups: mutants arrested at early stages and mutants with novel phenotypes. The members of the first group resemble wild-type proembryos and never reach other developmental stages. In the second group the tube-shaped mutants emb*-8522 and emb*-8535 completely lack apical-basal differentiation, while in mutant emb*-8516 a second embryo-like structure arises from the suspensor. The five emb mutations analyzed are non-allelic and two of the mutations are very likely caused by insertion of the transposon mutator, opening the door for their molecular analysis.

Journal Article↗

The development of M cells in Peyer's patches is restricted to specialized dome-associated crypts.

It is controversial whether the membranous (M) cells of the Peyer's patches represent a separate cell line or develop from enterocytes under the influence of lymphocytes on the domes. To answer this question, the crypts that produce the dome epithelial cells were studied and the distribution of M cells over the domes was determined in mice. The Ulex europaeus agglutinin was used to detect M cells in mouse Peyer's patches. Confocal microscopy with lectin-gold labeling on ultrathin sections, scanning electron microscopy, and laminin immuno-histochemistry were combined to characterize the cellular composition and the structure of the dome-associated crypts and the dome epithelium. In addition, the sites of lymphocyte invasion into the dome epithelium were studied after removal of the epithelium using scanning electron microscopy. The domes of Peyer's patches were supplied with epithelial cells that derived from two types of crypt: specialized dome-associated crypts and ordinary crypts differing not only in shape, size, and cellular composition but also in the presence of M cell precursors. When epithelial cells derived from ordinary crypts entered the domes, they formed converging radial strips devoid of M cells. In contrast to the M cells, the sites where lymphocytes invaded the dome epithelium were not arranged in radial strips, but randomly distributed over the domes. M cell development is restricted to specialized dome-associated crypts. Only dome epithelial cells that derive from these specialized crypts differentiate into M cells. It is concluded that M cells represent a separate cell line that is induced in the dome-associated crypts by still unknown, probably diffusible lymphoid factors.

Animals↗

Use of a digital film scanner to enhance low-power bright field photomicrography.

Low-power bright field photomicrographs often suffer from insufficient sharpness, uneven illumination, and colour hues. Using a film scanner, commercially available and designed for digitizing 35-mm transparencies, we directly scanned microscopic slides that carried dye-labelled and stained sections. The digital images covered a field of up to 24 x 36 mm and revealed excellent sharpness, absolutely even illumination and superior colour reproduction as compared to conventional photomicrographs taken with binoculars, macro lenses, or microscopes. As the method requires neither specialized instrumentation nor expert knowledge of photomicrographic techniques, it reduces costs and saves time. The high-quality digital survey micrographs can easily be used for image processing, image analysis and morphometry. Thus, this new method is valuable not only for pathology, embryology, histochemistry, and the neurosciences, but also for the exchange of low-power micrographs via the internet and for computer media that are increasingly used in medical education.

Image Enhancement↗