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Biomedical subjects

A Schmid

Publications and source records attributed to A Schmid.

At least 91 records · Page 5Linked to original sources

Clonal analysis of Drosophila embryonic neuroblasts: neural cell types, axon projections and muscle targets.

An experimental analysis of neurogenesis requires a detailed understanding of wild-type neural development. Recent DiI cell lineage studies have begun to elucidate the family of neurons and glia produced by each Drosophila embryonic neural precursor (neuroblast). Here we use DiI labeling to extend and clarify previous studies, but our analysis differs from previous studies in four major features: we analyze and compare lineages of every known embryonic neuroblast; we use an in vivo landmark (engrailed-GFP) to increase the accuracy of neuroblast identification; we use confocal fluorescence and Nomarski microscopy to collect three-dimensional data in living embryos simultaneously for each DiI-labeled clone, the engrailed-GFP landmark, and the entire CNS and muscle target field (Nomarski images); and finally, we analyze clones very late in embryonic development, which reveals novel cell types and axon/dendrite complexity. We identify the parental neuroblasts for all the cell types of the embryonic CNS: motoneurons, intersegmental interneurons, local interneurons, glia and neurosecretory cells (whose origins had never been determined). We identify muscle contacts for every thoracic and abdominal motoneuron at stage 17. We define the parental neuroblasts for neurons or glia expressing well-known molecular markers or neurotransmitters. We correlate Drosophila cell lineage data with information derived from other insects. In addition, we make the following novel conclusions: (1) neuroblasts at similar dorsoventral positions, but not anteroposterior positions, often generate similar cell lineages, and (2) neuroblasts at similar dorsoventral positions often produce the same motoneuron subtype: ventral neuroblasts typically generate motoneurons with dorsal muscle targets, while dorsal neuroblasts produce motoneurons with ventral muscle targets. Lineage data and movies can be found at http://www.biologists. com/Development/movies/dev8623.html http://www.neuro.uoregon. edu/doelab/lineages/

Animals↗

Mitochondria-derived and extra-mitochondrial human type-1 porin are identical as revealed by amino acid sequencing and electrophysiological characterisation.

In mammalian cells porin channels are localised in both mitochondrial outer membranes and extra-mitochondrial membranes. We isolated mitochondria-derived porin of a human lymphoblastoid B cell line, determined its amino acid sequence and characterised its channel properties. Interestingly, the amino acid sequence of this porin preparation and, correspondingly, its electrophysiological characteristics in a reconstituted system were identical to those of 'Porin 31HL', the human type-1 porin purified from a crude membrane preparation of the same cell line using a different purification protocol. The results raise questions about targeting, insertion and orientation of human type-1 porin in different membranes.

Amino Acid Sequence↗

Characterization of cell volume-sensitive chloride currents in freshly prepared and cultured pancreatic acinar cells from early postnatal rats.

1. In freshly prepared and cultured exocrine pancreatic acinar cells from 5- to 7-day-old rats a chloride-selective membrane conductance could be activated by intracellular application of GTPgammaS (40-100 microM), by application of positive pressure (5 cmH2O) to the pipette interior or by challenging the cells with a hyposmolar bath solution. Hyperosmolar bath solutions inhibited the cell volume-sensitive chloride currents. 2. The anion permeability sequence of the cell volume-sensitive chloride conductance was I- > Cl- approximately Br- > F- > methanesulphonate- > glutamate-. I- had a higher permeability but lower conductance than Cl-. The permeability ratio for Pglutamate/PCl was 0.12. 3. The cell volume-sensitive chloride conductance showed outward rectification. Membrane depolarization to high positive voltages (>= +60 mV) caused a time-dependent decay in outward currents. 4. DIDS (4, 4'-diisothiocyanatostilbene-2,2'-disulphonic acid) and SITS (4-acetamido-4'-isothiocyanatostilbene-2,2'-disulphonic acid) reversibly inhibited the cell volume-sensitive chloride current in a voltage-dependent manner. NPPB (5-nitro-2-(3-phenylpropylamino)-benzoic acid), quinidine, quinine and tamoxifen caused voltage-independent current inhibition. 5. Combined fura-2 and whole-cell current measurements showed that activation of the cell volume-sensitive chloride current does not involve cytosolic Ca2+ signals. Furthermore, there is no evidence that Ca2+-activated chloride currents play a significant role in cultured pancreatic acinar cells from 5- to 7-day-old rats. 6. Polymerase chain reaction followed by DNA sequence analysis indicated the presence of mRNA homologous to the ClC-3 chloride channel in pancreatic tissue from 5-day-old rats.

Animals↗

Medium chain length alkane solvent-cell transfer rates in two-liquid phase, pseudomonas oleovorans cultures

The oxidation of medium chain length alkanes and alkenes (C6 to C12) by Pseudomonas oleovorans and related, biocatalytically active recombinant organisms, in two-liquid phase cultures can be used for the biochemical production of several interesting fine chemicals. The volumetric productivities that can be attained in two-liquid phase systems can be, in contrast to aqueous fermentations, limited by the transport of substrates from an apolar phase to the cells residing in the aqueous phase and by toxic effects of apolar solvents on microbial cells. We have assessed the impact of these possible limitations on attainable productivities in two-liquid phase fermentations operated with mcl-alkanes. Pseudomonas oleovorans grows well in two-liquid phase media containing a bulk n-octane phase as the sole carbon source. However, cells are also damaged, typically resulting in a cell lysis rate of about 0.08 to 0. 10 h-1. These rates could be lowered by 50 to 70% to 0.03 h-1 and substrate yields increased from 0.55 to 0.85 g g-1 by diluting octane in non-metabolizable long-chain hydrocarbon solvents. Transfer rates of medium chain length (mcl) alkanes from the apolar phase to the cells were determined by following growth and the rate at which carbon-containing metabolites accumulated in the different phases of the cultures. mcl-Alkane solvent-cell transfer rates of at least 79, 64, and 18 mmol per liter of aqueous medium per hour were determined for n-heptane, n-octane, and n-decane, respectively. Rates of up to 30 mmol L-1 h-1 were observed under octane-limiting conditions in systems where the apolar substrate was dissolved to concentrations below 3% (v/v) in hexadecene. Based on low power input experiments, we estimated the maximum obtainable mass transfer rates in large scale processes to be in the range of 13 mmol L-1 h-1 for decane and higher than 45 mmol L-1 h-1 for octane and heptane. The results indicate that high solvent to cell mass transfer rates and minimized cell damage will enable high production rates in two-liquid phase bioprocesses, justifying ongoing efforts to attain high densities of catalytically, highly active cells in such systems. Copyright 1998 John Wiley & Sons, Inc.

Journal Article↗

LOE 908 blocks delayed rectifier type potassium channels in PC12 cells and cortical neurons in culture.

The effects of (R,S)-(3,4-dihydro-6,7-dimethoxy-isoquinoline-1-yl)-2- phenyl-N,N-di-[2-(2,3,4-trimethoxyphenyl)ethyl]-acetamide (LOE 908) were studied on K+ currents in undifferentiated cells from a phaeochromocytoma cell line (PC12), in cortical neurons from rat in primary culture, in a rat blood lymphoma cell line (RBL-1) and in a kidney cell line (BHK21). In PC12 cells delayed rectifier K+ currents measured in the whole-cell mode of the patch clamp technique were almost completely blocked by 10 microM LOE 908. The IC50 value was 0.7 microM and the Hill coefficient 0.8. After washout of the inhibitor about 80% of the current recovered. In rat cortical neurons in primary culture LOE 908 inhibited tetraethylammonium (TEA, 10 mM)-sensitive delayed rectifying K+ currents (LOE 908: 1 microM, 61 +/- 25% inhibition; 10 microM 103 +/- 19% inhibition). In contrast to the inhibitory action of LOE 908 on delayed rectifying K+ currents, Ca(2+)-activated potassium currents in BHK21 cells were only inhibited by 25 +/- 5% (10 microM LOE 908, n = 5) and no effect of LOE 908 was found on inward-rectifying K+ currents in RBL-1 cells. We conclude that LOE 908 is a K+ channel blocker with selectivity for delayed outward rectifying K+ channels.

Acetamides↗

Free plasma catecholamines in spinal cord injured persons with different injury levels at rest and during exercise.

Spinal cord lesion leads to an interruption of pathways from brain to the peripheral sympathetic nervous system, which results in pathological changes in sympathetic innervation. Free epinephrine (E), norepinephrine (NE) and dopamine (DA) were measured in 30 tetraplegics (TETRA), 15 high-lesion paraplegics (T1 and T4, HPARA), 15 paraplegics with injuries between T5 and T10 (MPARA), 15 low-lesion paraplegics (below T10, LPARA) and 16 non-handicapped control persons (C) at rest, at 60 and 100% of maximal oxygen consumption during graded wheelchair ergometry (WCE). The TETRA showed significant lower E and NE levels at rest and only slight increases during physical exercise. The E and NE concentrations of the paraplegics with a lesion below T5 were significantly higher than those of the high-lesion paraplegics, as well as those of the control persons at every point in the study. All paraplegics and the control persons showed, at submaximal and maximal exercise, a significant increase in NE. Only a slight increase in E in HAPRA was shown. No differences were found at rest and during exercise in E and NE levels in the MPARA and LPARA. No significant differences were found in the dopamine concentration at rest or during exercise for any of the groups. In summary, different levels of lesion and the resulting interruption to sympathetic pathways in the spinal cord are decisive factors in the degree of impairment of sympathetic innervation in SCI persons. Tetraplegics show less preganglionic resting activity because of interruption of impulses from central centers and no considerable stimulation of the sympathetic nervous system during maximal exercise. Individuals with high paraplegia have a partial impairment of catecholamine release, especially of epinephrine, at rest and during exercise. Paraplegics with a lesion level below T5 showed an augmented basal and exercise-induced upper spinal thoracic sympathetic activity in comparison to control persons.

Adult↗

Exposure of the surgeon to radiation during surgery.

Exposure to radiation over many years increases the incidence of cataracts and promotes the development of carcinoma of the thyroid gland. A prospective study of 24 operative procedures involving minimal invasive techniques and fluoroscopic guidance was undertaken in order to measure the radiation exposure to the primary surgeon. The study was conducted during 8 K-wire osteosyntheses in fractures of the distal radius, 8 closed interlocking intramedullary nailings in fractures of the femur and 8 internal fixator procedures, with or without posterior autogenic transpedicular bone grafting, in fractures of the lumbar spine. Radiation was monitored with the use of high sensitive thermoluminescent dosimeters. Fluoroscopy was necessary during the procedures, with exposure times ranging from 55 s to 12 min 35 s. The radiation dose received per procedure ranged from 0.6-259.3 microSv and was well within the dose limits set by German law.

Fluoroscopy↗

Degradation of and intraosseous reactions to biodegradable poly-L-lactide screws: a study in minipigs.

In 20 minipigs an osteotomy of the lumbar spine was done at L3 and fixed with resorbable implants (plate and screws) made of poly-L-lactide. For 24 months postoperatively the cancellous bone surrounding the screws was examined histologically. Specimens from the body of lumbar spine L2 and L4, carrying the screws, were fixed in a series of ethanol immersions and then embedded in methylmethacrylate. Longitudinal and transverse sections were cut with a microtome at 4-5 microns and stained by the Masson-Goldner and Giemsa method for light microscopy. Our investigation confirmed a long degradation period of poly-L-lactide and little inflammatory reaction within the bone during the whole period of observation. For poly-L-lactide the degradation time exceeded the longest follow-up time in this study, 24 months. Polymorphonuclear granulocytes and mononuclear round cells were scarce in all follow-up groups. Osteolytic changes or the emergence of seroma was not observed. Textured bone was evident early close to a thin fibrous capsule surrounding the screws. The remodelling of the bone close to the screws ensues by the formation of osteons.

Absorbable Implants↗

Heart rate deflection compared to 4 mmol x l(-1) lactate threshold during incremental exercise and to lactate during steady-state exercise on an arm-cranking ergometer in paraplegic athletes.

The deflection point (DP) of the heart rate in relation to the work rate (WR) of 8 male endurance-trained paraplegics and 11 male physically active sports students was investigated during nonsteady-state incremental arm cranking ergometry (IT) and compared to the 4 mmol x l(-1) blood lactate concentration threshold and to blood lactate concentration in steady-state exercise (SST). Heart rate, and lactate concentration from capillary blood, were determined at rest, during IT and SST. The DP was calculated by linear regression analysis of the heart rate during IT. The SST consisted of three consecutive exercise intensities over a period of 8 min at exercise intensities of 10 W below, and at 10 W above the work rate at deflection point (WRDP). No difference was found between the paraplegics and non-handicapped subjects regarding heart rate and blood lactate concentration at rest and during exercise. A DP was established in all the paraplegics and in 72.7% of the non-handicapped subjects, but lactate accumulation was observed in 75% of the paraplegics and in 62.5% of the non-handicapped subjects at the lowest intensity of SST. In summary, endurance-trained paraplegics with an injury level below T5 showed heart rate and blood lactate concentration values comparable to non-handicapped subjects during IT. A linear increase at moderate exercise intensities and a levelling-off at higher to maximal intensities could be identified in all the paraplegics and in 72.7% of non-handicapped subjects. The determination of the anaerobic threshold by DP should be applied with caution, since no causal relationship of DP and the anaerobic threshold was found and the WRDP tended to overestimate threshold values.

Adult↗

Developments toward large-scale bacterial bioprocesses in the presence of bulk amounts of organic solvents.

Many pseudomonads and other bacteria can grow on aliphatic and aromatic hydrocarbons that occur in the environment. We are examining the potential of such organisms as biocatalysts for the oxidation of a variety of substituted aliphatic and aromatic compounds. To attain a high production rate of oxidation products via such biotransformations, we have focused on two-liquid phase culture systems. In these systems, cells are grown in liquid media consisting of an aqueous phase containing water-soluble growth substrates and droplets of a water-immicible organic solvent containing bioconversion substrates and products. For industrial applications of such two-liquid phase processes, several questions remain. What are the maximum rates at which apolar compounds can be transferred from the apolar phase to cells growing in the aqueous phase, i.e., what are the maximum space-time yields attainable in two-liquid phase fermentations under practical conditions? What does an efficient downstream processing of two-liquid phase medium involve? What safety regimes should be considered in working with flammable organic solvents? Can elevated pressure be used to increase oxygen transfer? Based on answers to these questions, we have recently developed a high-pressure, explosion-proof bioreactor system with Bioengineering AG (Wald, Switzerland), which will be installed in our pilot plant and used to explore two-liquid phase bioconversions at a pilot scale.

Bacteria↗

Endogenous B1 receptor mediated hypotension produced by contact system activation in the presence of endotoxemia.

Previous studies have shown that an intravenous infusion of dextran sulfate (DXS) causes arterial hypotension via release of bradykinin (BK) and stimulation of bradykinin B2 receptors in pigs. The bradykinin B1 receptor is not physiologically present but its expression can be induced by bacterial lipopolysaccharide (LPS). This study was designed to assess the relative roles of bradykinin B2 and B1 receptors in the hypotensive response produced by DXS in LPS-treated pigs. In LPS-treated pigs a continuous infusion of DXS produced a progressive drop in blood pressure that peaked at approximately 30 min after onset of the infusion and returned to baseline after another 30 min. In animals receiving the selective B2 receptor antagonist Hoe-140 a significant attenuation of the peak fall in blood pressure to DXS was observed. In pigs treated with Hoe-140 and the selective B1 receptor antagonist CP-0298 (Lys(0)-Leu(8)-des-Arg(9)-bradykinin) DXS infusion had no effect on blood pressure. This is the first demonstration in vivo that following activation of the contact system both B2 and B1 receptors are involved in the resulting hypotensive response. This would be consistent with the production of BK (which stimulates B2 receptors) that is subsequently converted to the biologically active metabolite des-Arg(9)-BK in sufficient concentrations to activate B 1 receptors. The significance of these observations to pathophysiology remains to be determined.

Adrenergic beta-Antagonists↗

Assessment of carotid wall motion and stiffness with tissue Doppler imaging.

In this study, the wall motion of the common carotid artery was characterized by measuring wall-motion velocity (Wv) with tissue Doppler imaging (TDI) in 78 male and female subjects (16-75 y) with no history of cardiovascular disease. The near and far arterial wall showed essentially different Wv patterns. To assess the vascular systolic distension, the Wv of the near and far arterial walls were measured and a Wv index was calculated by subtracting the far Wv from the near Wv. Aging was associated with a 2.0-2.5-fold decrease of peak Wv index. Corrected for carotid diameter and blood pressure, the peak Wv index and mean systolic acceleration to the peak Wv correlated highly with arterial distensibility (r = 0.81 resp. r = 0.75) and compliance (r = 0.73 resp. r = 0.68). This study demonstrates the feasibility of TDI in the characterization of wall motion patterns and in the assessment of common carotid artery stiffness.

Adolescent↗

Absence of Gcn5 HAT activity defines a novel state in the opening of chromatin at the PHO5 promoter in yeast.

Histone acetyltransferase (HAT) activity has been demonstrated for several transcriptional activators, formally connecting chromatin modification with gene regulation. However, no effect on chromatin has been demonstrated. We have investigated the role of the HAT Gcn5 at the nucleosomally regulated PHO5 promoter. Under conditions of constitutive submaximal activation (i.e., in the absence of the negative regulator Pho80), deletion of Gcn5 determines a novel randomized nucleosomal organization across the promoter and leads to a dramatic reduction in activity. Furthermore, mutation of amino acids critical for Gcn5 HAT activity is sufficient to generate this structure. This intermediate state in chromatin opening gives way to the fully open structure upon maximal induction (phosphate starvation), even in the absence of Gcn5. Thus, Gcn5 is shown to affect directly the remodeling of chromatin in vivo.

Acetyltransferases↗

Autoantibodies to C-reactive protein (CRP) and other acute-phase proteins in systemic autoimmune diseases.

Autoantibodies to CRP were reported previously in patients suffering from toxic oil syndrome. This syndrome resembles autoimmune diseases such as systemic lupus erythematosus (SLE) or systemic scleroderma. We therefore examined the prevalence of antibodies to CRP and other acute-phase proteins in autoimmune diseases, including SLE, subacute cutaneous lupus erythematosus (SCLE), systemic scleroderma (SSc), and primary biliary cirrhosis (PBC), as well as in bone marrow transplantation-induced chronic graft-versus-host disease and eosinophilia-myalgia syndrome. IgG antibodies to CRP were found in 78% of SLE and in 30% of SCLE patients, while 16% of patients with PBC were positive. In up to 45% of patients with SSc predominantly IgG antibodies to ceruloplasmin were detectable. Lack of systemic involvement as in discoid lupus erythematosus and localized scleroderma (morphea) correlated with low or absent antibody formation. However, no correlation was found between anti-acute-phase protein antibodies with liver disease or other organ involvement. Adsorption studies revealed that non-native epitopes on the CRP molecule, termed modified CRP, are the main target of antibodies. Chronic inflammatory tissue injury in systemic autoimmune disease might increase the presentation of cryptic epitopes of CRP to the threshold required for T cell activation.

Acute-Phase Proteins↗

[Injury risk of competitive, handicapped cross-country skiers in training nd competition].

Injuries caused by cross country skiing have been poorly investigated in handicapped athletes. The dynamic sliding shape of motion makes this sport to a suitable discipline for people with a deficit of locomotion. Visual handicapped people with a guide are able to improve their motoric skills, co-ordination, orientation and body self-consciousness in the track. Since handicapped athletes are performing in international competitions the training intensity to fulfill the requirements, but also the risk of overstrain induced injuries got increased, like in other high-performance sports. Our study examined injuries and overuse syndromes of the German National Team Ski Nordic during the Paralympics in Tignes/ Albertville (1992). Lillehammer (1994) and the training period in preparation for the Paralympics in Nagano (March 1998). The incidence and kind of injuries in the competitive handicapped cross country skier was comparable with non-handicapped athletes, but the injury pattern was different.

Adult↗

Cardiovascular differences between sedentary and wheelchair-trained subjects with paraplegia.

PURPOSE: Heart dimensions, left ventricular function, and internal dimensions of limb arteries, as well as physical fitness, were examined in sedentary male subjects with paraplegia (SP, N = 20), national elite male athletes with paraplegia (PA, N = 29), and untrained able-bodied males (AB, N = 30). METHODS: All subjects underwent two-dimensional echocardiography, duplex sonography of common femoral artery and subclavian artery at rest, and an incremental wheelchair ergometer exercise test. RESULTS: Heart volume in relation to body weight was not different in PA as compared with that in AB (11.5 +/- 1.6 vs 11.6 +/- 2.2 mL.kg-1; mean +/- SD), whereas SP showed significantly lower values (9.7 +/- 1.5 mL.kg-1). Left ventricular ejection fraction was similar in all subjects (59.9-60.8%). In relation to body surface area, subclavian artery cross-sectional area was significantly higher in PA compared with that in AB and SP, respectively (PA: 0.32 +/- 0.05, AB: 0.21 +/- 0.06, SP: 0.22 +/- 0.05 cm2/m2). The corresponding values for the common femoral artery were significantly lower in all subjects with paraplegia as compared with those in AB, whereas no difference was found between PA and SP (AB: 0.31 +/- 0.05, PA: 0.14 +/- 0.05, SP: 0.15 +/- 0.04 cm2/m2). Peak oxygen uptake (VO2peak) determined in the wheelchair ergometer exercise test was within the same range in PA and AB, but significantly (P < 0.05) lower in SP (PA: 34.5 +/- 4.3, AB: 31.5 +/- 4.1, SP: 23.9 +/- 3.8 mL.kg-1.min-1). CONCLUSIONS: In conclusion, cardiac dimensions and VO2peak of PA were larger than in SP but do not exceed those of AB. Intensive wheelchair training was associated with larger dimensions of the subclavian arteries in PA, whereas a hypotrophy of the common femoral artery was found in SP and PA compared with that in AB.

Adult↗

Towards a biocatalyst for (S)-styrene oxide production: characterization of the styrene degradation pathway of Pseudomonas sp. strain VLB120.

In order to design a biocatalyst for the production of optically pure styrene oxide, an important building block in organic synthesis, the metabolic pathway and molecular biology of styrene degradation in Pseudomonas sp. strain VLB120 was investigated. A 5.7-kb XhoI fragment, which contained on the same strand of DNA six genes involved in styrene degradation, was isolated from a gene library of this organism in Escherichia coli by screening for indigo formation. T7 RNA polymerase expression experiments indicated that this fragment coded for at least five complete polypeptides, StyRABCD, corresponding to five of the six genes. The first two genes encoded the potential carboxy-terminal part of a sensor, named StySc, and the complete response regulator StyR. Fusion of the putative styAp promoter to a lacZ reporter indicated that StySc and StyR together regulate expression of the structural genes at the transcriptional level. Expression of styScR also alleviated a block that prevented translation of styA mRNA when a heterologous promoter was used. The structural genes styA and styB produced a styrene monooxygenase that converted styrene to styrene oxide, which was then converted to phenylacetaldehyde by StyC. Sequence homology analysis of StyD indicated a probable function as a phenylacetaldehyde dehydrogenase. To assess the usefulness of the enzymes for the production of enantiomerically pure styrene oxide, we investigated the enantiospecificities of the reactions involved. Kinetic resolution of racemic styrene oxide by styrene oxide isomerase was studied with E. coli recombinants carrying styC, which converted styrene oxide at a very high rate but with only a slight preference for the S enantiomer. However, recombinants producing styrene monooxygenase catalyzed the formation of (S)-styrene oxide from inexpensive styrene with an excellent enantiomeric excess of more than 99% at rates up to 180 U g (dry weight) of cells-1.

Base Sequence↗