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Biomedical subjects

S König

Publications and source records attributed to S König.

At least 19 recordsLinked to original sources

Use of repeated measures analysis for evaluation of genetic background of dairy cattle behavior in automatic milking systems.

Milking frequencies measured at official test days were used with repeated measurement analysis to reveal the environmental and genetic impact on the milking frequency of cows in automatic milking systems. Repeated measurements were 3 test-day observations per cow within days in milk (DIM) classes, with 1,216 cows in DIM class 1 (d 0 to 99), from 1,112 cows in DIM class 2 (d 100 to 199), and from 1,004 cows in DIM class 3 (d 200 to 299) kept in 15 farms. Selection criteria for models analyzing repeated measurements were Akaike and Schwarz Bayesian values, which favored the autoregressive [AR(1)] covariance structure over the compound symmetry model. Results from the AR(1) model indicated a significant impact of fixed herd and parity effects. Milking frequencies decreased with increasing parities and were greatest for first-parity cows. High daily milk yield was associated with higher milking frequencies. Heritabilities for milking frequency were 0.16, 0.19, and 0.22 in DIM classes 1, 2, and 3, respectively, from the AR(1) model. Higher heritabilities in the later stage of lactation were due to a substantial reduction of the residual variance. Genetic correlations between test-day milk yield and daily milking frequency were in the range of 0.46 to 0.57 for all DIM classes and between milking frequency and somatic cell score were near zero. For verification of results, milking frequencies of the same cows obtained from herd management programs were averaged within DIM classes. Heritabilities were slightly above the values from the AR(1) model. In conclusion, heritabilities for milking frequency in automatic milking systems are moderate enough to incorporate this behavioral trait in a combined breeding goal. The inevitable improvement of labor efficiency in dairy cattle farming demands such cows going easily and voluntarily in automatic milking systems.

Analysis of Variance↗

[Role of stem cells in adult hepatic regeneration].

Central to the successful surgically treatment numerous liver diseases is the ability of the organ to regenerate. The understanding of the process of self-renewal has both changed and progressed over the last few decades. For many years, the assumption was that the liver regenerates primarily through the division of mature liver cells. However, over the last few years there has been increasing evidence of the participation of stem cells. Intrahepatic stem cells, so-called oval cells, are activated under conditions of severe or chronic liver disease and originate from the canals of Hering. In addition, extrahepatic stem cells may migrate from the bone marrow into the liver when the regenerative capacity of the liver itself is depleted. It is not yet fully clear how the different stem cell populations interact with both each other and the mature liver cell population to achieve homeostatic cell and differentiation equilibrium in the diseased and/or regenerating organ. In any case, the outstanding growth potential of liver stem cells may become a clinically viable option in the field of cell transplantation.

Adult↗

Genetic relationships for dairy performance between large-scale and small-scale farm conditions.

Genotype by environment interaction can be detected via the estimation of genetic correlations between environments under an animal model based on data comprising genetic links between the strata. Genetic correlations were estimated for protein yield of Holstein cows within and across regions of Germany using REML under an animal model for lactation and test-day records. Subsets of the entire data were created, stratified by region or herd size within region, and comprised between 16,307 and 132,972 cows with first-lactation records. Substantial heterogeneity exists between regions in Western and Eastern Germany. In Western states, most farms are small, with typical herd sizes of 30 to 60 cows, whereas in Eastern states, mostly large herds with herd sizes of 500 to 2000 cows are common. The results show drastic differences for residual and permanent environmental variance components between Eastern and Western regions with increases of around 30% for Eastern regions. Additive genetic variances were of similar magnitude in both regions. Genetic correlations between Eastern and Western states were between 0.90 and 0.95 but dropped to 0.79 when data from an Eastern state were reduced to contain large herds only. The results indicate that differences in herd size account for more of the differences in genetic correlation than do geographic regional differences.

Animals↗

Metabolism of MDCK cells during cell growth and influenza virus production in large-scale microcarrier culture.

The production of equine influenza in Madin-Darby canine kidney (MDCK) cells in large-scale microcarrier culture is described with detailed on- and off-line analytical data during cell growth and virus replication. Metabolite concentration profiles for glucose, glutamine, lactate and ammonium are shown. Lactate and ammonium concentrations were always below inhibiting levels. Concentration profiles for essential and non-essential amino acids of the cell culture medium are discussed. During cell growth proline was released into the medium with a significant rate while two amino acids, serine and methionine were almost depleted. After infection, virus titer increased after a delay of 10-16 h whereas first changes in amino acid metabolism could be observed within 4h post-infection. Here, glutamate and aspartate increase correlated to virus release kinetics, indicating cell disruption and apoptosis. Starting with a moi of 0.025 resulted in a maximum virus yield of 2.4 log HA/100 microl at 44 h post-infection.

Amino Acids↗

Method for isolating tight-binding inhibitors of rat lens aldose reductase.

Numerous animal studies indicate that aldose reductase inhibitors (ARIs) are beneficial for the prevention or amelioration of diabetic complications such as neuropathy, nephropathy and the ocular complications of cataract, retinopathy and keratopathy. To aid in the identification of novel potent ARIs, we have previously developed a screening method that is based on the formation of a non-covalent ternary tight-binding enzyme-inhibitor-nucleotide (AR-ARI-NADPH) complex that can be isolated using YM-10 filter units. Here, we report a modification of this method that permits us to rapidly identify tight binding ARIs that are isolated by denaturation from AR-ARI-NADPH complexes that are free of possible contamination resulting from the reaction of methanol with the YM-10 filter units. For the development of this procedure, nine structurally diverse ARIs were mixed with purified recombinant rat lens aldose reductase (RLAR) bound with NADPH to form tight-binding RLAR-ARI-NADPH complexes. These complexes were purified by high pressure Sephadex 75 size exclusion chromatography using ammonium acetate buffer and the formation of each complex was confirmed by electrospray ionisation mass spectrometry (ESI-MS). Each of the complexes was then denatured with methanol, rechromatographed on the size exclusion column, and the identity of the bound ARIs was confirmed by ESI-MS. The apparent ARI binding with aldose reductase to form a tight binding ARI complex appeared proportional to their IC50 values. This procedure allows for the rapid identification of tight binding ARIs with apparent IC50s<0.1 microm.

Aldehyde Reductase↗

Myelinopathia centralis diffusa (vanishing white matter disease) in a four-year-old boy.

A four-year-old boy presented with moderate ataxia triggered by a minor head trauma several weeks ago. Discrepantly severe signal changes of cerebral white matter with almost CSF-isointense signal on all pulse sequences were detected at cranial MRI. Localized proton MR spectroscopy of cerebral white matter demonstrated an even decrease of all metabolites. Glycine was found elevated in CSF. This pattern of clinical history, MR imaging and spectroscopy features and elevated glycine in CSF is characteristic for a novel entity amongst the leukoencephalopathies of childhood. It was originally termed "myelinopathia centralis diffusa" and renamed "vanishing white matter disease" later.

Brain Diseases↗

Live-wires using path-graphs.

OBJECTIVES: This article discusses our new Path-Graph approach for the interactive Live-Wire segmentation method in 2D applied to pre-segmented data. Furthermore, we examine whether or not the Live-Lane extension provides advantages in combination with pre-segmentation. METHODS: We automatically over-segment the image data in a preprocessing step, using region growing with an automatic seed point generation. The Live-Wire algorithm is applied on this mosaic data by using the outlines of the homogeneous regions as the basis for graph building. We present a new definition of this underlying graph where the edges of the standard graphs are turning into vertices and the vertices of the new graph are defined by the edge connectivity in the standard graph. For better differentiation we name our new graph Path-Graph and the original defined graph Node-Graph. RESULTS: The quality evaluation is done by comparing our segmentation results with existing model data. We show that using the Path-Graph as basis for the Live-Wire algorithm instead of the Node-Graph allows for a finer segmentation. We achieve a reduction of incorrectly classified pixels by 20.66 per cent and a decrease of the mean boundary deviation by 11.61 per cent. Since savings on cost tree calculations are compensated by additional computation time required to compute the Live-Lanes, a performance loss of 2.41 per cent is measured. CONCLUSIONS: Our redefinition of the underlying graph increases the quality of the Live-Wire segmentation. The Live-Lane extension in combination with pre-segmentation is not justified for our data.

Algorithms↗

[Rehabilitation of extensor tendon injuries].

The extensor mechanism of the hand consists of different tendons, originating from extrinsic and intrinsic muscles of the hand and forearm. Because of their course near to skin and bone separated only by a thin layer of paratenon, they are very sensitive to injuries and adhesions. Treatment protocols have to be adapted according to the different zones. This paper explains the basic principles of rehabilitation in extensor tendon injuries.

Aftercare↗

Stability of bone grafting and placement of implants in the severely atrophic maxilla.

A severely atrophic maxilla can be restored by bone grafts to allow the insertion of implants. We present 30 consecutive patients treated with autogenous inlay and onlay bone grafts from the iliac crest to the floor of the maxillary sinus and the alveolar crest. A total of 200 implants were inserted 4-6 months after bone grafting. A mean vertical increase in bone thickness of 14mm was achieved. After a mean bone loss of 1.3mm during the first year after bone grafting only minimal resorption was observed during the second and third year. Seven implants failed to integrate and a further four implants were lost during follow-up.

Alveolar Bone Loss↗

Isolation of total-RNA from formalin-fixed rat retina.

The aim of this project was to establish a method for the purification of total-RNA from fixed rat-retina. Two different established methods were used for RNA purification, and successful isolation was verified with RT-PCR for amplification of beta-actin (two different product-lengths) and subsequent gel-electrophoresis. Total-RNA was successfully isolated from fixed rat-retina. The house keeping gene, beta-actin could be detected after fixing the retina either with 1% formalin or with 4% paraformaldehyde (PFA). Hexamer-primer based RT-PCR gave better results than the oligo-d(T)-primer based RT-PCR method. Both the 698 and 225 bp beta-actin-fragments could be successfully amplified, where amplification of the latter was more efficient. This approach shows that tissue fixation prior to RNA-isolation facilitates the rapid isolation of undamaged RNAs in tissues such as the retina, which are known to yield low levels of RNA and are vulnerable to RNases.

Animals↗

Design, formation and properties of tetrahedral M(4)L(4) and M(4)L(6) supramolecular clusters.

The rigid tris- and bis(catecholamide) ligands H(6)A, H(4)B and H(4)C form tetrahedral clusters of the type M(4)L(4) and M(4)L(6) through self-assembly reactions with tri- and tetravalent metal ions such as Ga(III), Fe(III), Ti(IV) and Sn(IV). General design principles for the synthesis of such clusters are presented with an emphasis on geometric requirements and kinetic and thermodynamic considerations. The solution and solid-state characterization of these complexes is presented, and their dynamic solution behavior is described. The tris-catecholamide H(6)A forms M(4)L(4) tetrahedra with Ga(III), Ti(IV), and Sn(IV); (Et(3)N)(8)[Ti(4)A(4)] crystallizes in R3(-)c (No. 167), with a = 22.6143(5) A, c = 106.038(2) A. The cluster is a racemic mixture of homoconfigurational tetrahedra (all Delta or all Lambda at the metal centers within a given cluster). Though the synthetic procedure for synthesis of the cluster is markedly metal-dependent, extensive electrospray mass spectrometry investigations show that the M(4)A(4) (M = Ga(III), Ti(IV), and Sn(IV)) clusters are remarkably stable once formed. Two approaches are presented for the formation of M(4)L(6) tetrahedral clusters. Of the bis(catecholamide) ligands, H(4)B forms an M(4)L(6) tetrahedron (M = Ga(III)) based on an "edge-on" design, while H(4)C forms an M(4)L(6) tetrahedron (M = Ga(III), Fe(III)) based on a "face-on" strategy. K(5)[Et(4)N](7)[Fe(4)C(6)] crystallizes in I43(-)d (No. 220) with a = 43.706(8) A. This M(4)L(6) tetrahedral cluster is also a racemic mixture of homoconfigurational tetrahedra and has a cavity large enough to encapsulate a molecule of Et(4)N(+). This host-guest interaction is maintained in solution as revealed by NMR investigations of the Ga(III) complex.

Algorithms↗

Effect of L-carnitine treatment for valproate-induced hepatotoxicity.

The authors analyzed the association of L-carnitine treatment with hepatic survival in 92 patients with severe, symptomatic, valproate-induced hepatotoxicity. Forty-eight percent of the 42 patients treated with L-carnitine survived, but only 10% of the 50 patients treated solely with aggressive supportive care survived (p < 0.001). Early intervention with IV rather than enteral L-carnitine was associated with the greatest hepatic survival. Specifically, all 10 patients who were diagnosed in <5 days and treated with IV L-carnitine survived. Most patients had features of chronic illness and most children appeared to be malnourished.

Adolescent↗

Examination of donor substrate conversion in yeast transketolase.

The cleavage of the donor substrate d-xylulose 5-phosphate by wild-type and H263A mutant yeast transketolase was studied using enzyme kinetics and circular dichroism spectroscopy. The enzymes are able to catalyze the cleavage of donor substrates, the first half-reaction, even in the absence of any acceptor substrate yielding d-glyceraldehyde 3-phosphate as measured in the coupled optical test according to Kochetov (Kochetov, G. A. (1982) Methods Enzymol. 90, 209-223) and compared with the H263A variant. Overall, the H263A mutant enzyme is less active than the wild-type. However, an increase in the rate constant of the release of the enzyme-bound glycolyl moiety was observed and related to a stabilization of the "active glycolaldehyde" (alpha-carbanion) by histidine 263. Chemically synthesized dl-(alpha,beta-dihydroxyethyl)thiamin diphosphate is bound to wild-type transketolase with an apparent K(D) of 4.3 +/- 0.8 microm (racemate) calculated from titration experiments using circular dichroism spectroscopy. Both enantiomers are cleaved by the enzyme at different rates. In contrast to the enzyme-generated alpha-carbanion of (alpha,beta-dihydroxyethyl)thiamin diphosphate formed by decarboxylation of hydroxylactylthiamin diphosphate after incubation of transketolase with beta-hydroxypyruvate, the synthesized dl-(alpha,beta-dihydroxyethyl)thiamin diphosphate did not work as donor substrate when erythrose 4-phosphate is used as acceptor substrate in the coupled enzymatic test according to Sprenger (Sprenger, G. A., Schörken, U., Sprenger, G., and Sahm, H. (1995) Eur. J. Biochem. 230, 525-532).

Amino Acid Substitution↗

[Teaching and learning in surgery--The Gottingen curriculum].

The study of medicine in Germany is in need of reform. Oversubscribed courses, the lack of practical reference, scarce patient contact and cancelled lessons define the educational landscape for many students. Since at present we cannot carry out global reforms, the quality of medical education is highly dependent on the commitment of those responsible in the individual institution. The aim of the revised surgical curriculum in Göttingen is to demonstrate how medical education with a high didactic level including multimedia teaching can be realised despite large student numbers. Duties both in patient care and education are coordinated through integrated organisation. Educational content and structure are made transparent by online guidebooks. In the surgical examination course and the practical course in surgery, patient-oriented tuition takes place consistently in small groups. The Teaching Studio provides additional aids such as interactive CD-ROMs, online research, textbooks, ultrasound equipment, examination and suturing dummies. Curriculum-relevant information, including the weekly case examples and instruction guides for clinical practical experience, is presented on an Internet website. Constant evaluation of the curriculum not only documents student motivation, but also serves to improve the educational concepts on a continuous basis. On completion of the practical course in surgery, 65% of participants maintain that they have reached the defined learning objectives "well" or "very well" (prior to reform only 17%). In the summer semester 2000, 46% of students gave top marks in the practical course in surgery with respect to tuition in patient contact. The results of the evaluations verify the positive feedback from the student body.

Clinical Competence↗

Children with epilepsy after withdrawal of anticonvulsive therapy: psychiatric behaviour and neuropsychological abilities.

UNLABELLED: Do children who remain in remission with epilepsy after withdrawal of anticonvulsive therapy still exhibit a different behaviour and performance-related abilities compared to healthy children? It was the aim of this study to investigate whether these children showed deviating psychiatric behaviour and/or neuropsychological function deficits as compared with a healthy control group and with children following occasional seizures which had never been treated. The study comprised 39 children with epilepsy after withdrawal of anticonvulsive therapy, 37 children with occasional seizures, and 39 matched healthy control children who were tested with regard to psychiatric and neuropsychological criteria. The patients were aged 6-15 years; 28 had a primarily generalised epilepsy, and 11 had a focal epilepsy. The patients remained free from seizures for more than 1 year. Psychiatric testing comprised the Child Behavior Checklist by Achenbach and Edelbrock and the Mannheim Parent Interview. Neuropsychological testing included non-verbal intelligence, visual perception, short-term memory and learning ability, arithmetics, movement coordination and motor skills, fine motor skills and verbal functions. After withdrawal from anticonvulsive therapy, the children with epilepsy did not exhibit any psychiatric disturbances, whereas those with occasional seizures frequently showed behaviour problems. After withdrawal from anticonvulsive therapy, no neuropsychological deficits were found in the children with epilepsy as compared to those with occasional seizures or to healthy children. Risk factors for poorer test results were first seizures occurring before the age of 3 years, delayed development, and neonatal problems. CONCLUSION: Children with epilepsy who remained without relapse after withdrawal from the anticonvulsive therapy did not show any psychiatric abnormalities and their performance was comparable to that of healthy control children, provided no risk factors were present.

Adolescent↗

Phosphorylation by PKA of a site unique to B-Raf kinase.

The Raf kinases serve as central intermediates to relay signals from Ras to ERK. Cell-specific effects of these signals on growth, differentiation and survival can be observed due to the recruitment of different isoenzymes of the Raf family. The in vitro phosphorylation of a site unique to B-Raf (Ser429) has been proposed to be responsible for the negative regulation of the isoenzyme by Akt. Using phosphopetide mapping and site-directed mutagenesis we showed that Ser429 is phosphorylated upon cAMP elevation in PC12 cells and proposed that PKA is a major kinase phosphorylating the B-Raf-specific site in vivo.

Amino Acid Sequence↗

Use of nonspecific cleavage products for protein sequence analysis as shown on calcyclin isolated from human granulocytes.

In this paper, analysis strategies developed for a sequencing problem concerning the identification of an S100 protein isolated from human granulocytes are discussed. The analysis of a trypsinized lyophilized sample suggested the presence of a number of peptides which are non-tryptic in origin. During purification of proteins from cell lysates nonspecific cleavage can be observed which may reflect biological processes and can become an unavoidable analytical problem. Current mass spectrometric software is evaluated for the analysis of nonspecific digests in this context. Matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), high-performance liquid chromatography (HPLC)-MS/MS, and selected ion monitoring (SIM)-MS/MS have been used for peptide analysis and in addition HPLC-MS was carried out for protein analysis leading to the detection of an N-terminal modification of the protein. The success of the study is mainly due to the careful investigation of nonspecific cleavage products. Data obtained from the routine mass spectrometric analysis of an in-gel-digest allowed the identification of this protein as S100 calcium-binding protein A6-calcyclin whose expression in granulocytes has not been described so far.

Amino Acid Sequence↗