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K Mewes

Publications and source records attributed to K Mewes.

14 recordsLinked to original sources
PubMed1994

Enrichment of bovine X and Y spermatozoa by free-flow electrophoresis.

Conditions of separation of bovine spermatozoa according to their differences in electrophoretic mobility were studied using free-flow electrophoresis. Distribution of spermatozoa in the electric field depends on composition of buffer system, field intensities and temperature. Conditions are described for reproducible separation of anodically migrating spermatozoa into two distinct peaks both for cold-immobilized (10-13 degrees C) and for motile cells (25 degrees C). Electrophoresis with the buffer used at 25 degrees C and field intensities of 70-100 V/cm provided high percentages of motile spermatozoa (50-90%) in all fractions. However, the sperm distribution across the fractions was the result of superimposed passive electrophoretic motion towards the anode and the active galvanotactic migration towards the cathode. Separation of X- and Y-spermatozoa was verified by quantification of Y-spermatozoa by means of both in situ hybridization, using Y-specific DNA fragments, and the Y-specific fluorescence staining (f-body test). Y-spermatozoa could be enriched in anodic fractions (112 and 144% relative to control). X-spermatozoa were enriched in cathodic fractions (80 and 68% Y-spermatozoa, as compared to control).

Animals↗
PubMed1994

Primate rubromotoneuronal cells: parametric relations and contribution to wrist movement.

1. Fifty-nine rubromotoneuronal (RM) cells were identified in two rhesus monkeys on the basis of their postspike facilitation (PSpF) of rectified electromyographic (EMG) activity. These cells were studied in relation to a step tracking task requiring wrist movements between fixed target zones in flexion and extension. Movement away from a 0 position was opposed by spring-like loads (auxotonic). Additionally, nine cells were evaluated using an isometric task. Neuronal discharge could be divided into three basic components: background discharge in the absence of movement, phasic modulation during movement, and tonic modulation during sustained holding against external loads. 2. Four basic patterns of RM cell activity were observed in relation to ramp-and-hold wrist movements: phasic-tonic (44%), pure phasic (22%), pure tonic (2%), and unmodulated (24%). The discharge of unmodulated cells did not covary with movement parameters but, as with other RM cells, background discharge did increase in association with task performance. 3. The phasic discharge of RM cells led to the onset of target muscle EMG activity by an average of 89 +/- 82 ms (mean +/- SD, n = 104) in extensors and 88 +/- 74 ms (n = 30) in flexors. Target muscles are defined as ones showing PSpF of EMG activity. It was found that 94% of extensor and 87% of flexor RM cells discharged before or synchronous with the onset of target muscle EMG activity. 4. Thirty-one RM cells (53%) showed a tonic increase in cell activity during the static hold phase of the task. Twenty-three of these were tested for relations to static torque. Fifteen extension related cells and one flexion cell had significant, positive regression slopes for the relation between tonic discharge rate and static torque. The mean rate-torque slope for extension related cells was 160 Hz/Nm and 103 Hz/Nm for flexion related cells. These mean slopes are about one-third those of corticomotoneuronal (CM) cells. 5. Cell discharge rate was correlated with velocity and rate of change of torque (dT/dt) for 32 RM cells with a phasic component of discharge during movement. The peak increase in phasic discharge above tonic firing rate (PDI, peak dynamic index) was significantly correlated only with velocity in eight cells and only with dT/dt in five cells. The phasic discharge of four additional cells was correlated with both velocity and dT/dt, but for three of these cells, the correlation was stronger for velocity. The mean slope for the relation between velocity and PDI was 0.31 Hz.deg-1.s-1.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1994

Synthesis and structural elucidation of biliary excreted thioether derivatives of mitoxantrone.

1. Hplc-MS coupling has been used for the identification of thioether derivatives of the anticancer agent mitoxantrone in the bile of pig. 2. Three biologically relevant new thioether derivatives of mitoxantrone have been synthesized by a horseradish peroxidase-catalysed reaction. 3. The thioether derivatives have been characterized by means of ion-spray tandem mass spectrometry and nmr spectrometry including two-dimensional techniques. 4. The carbon-sulphur bond formation takes place at the hydroquinone moiety of the anthraquinone skeleton pointing to the importance of a tautomeric equilibrium between different species of the oxidized drug. 5. The occurrence of the synthesized compounds in biological systems suggests a metabolic pathway that may be relevant for the cytotoxicity of mitoxantrone (oxidative activation).

Animals↗
PubMed1994

Busulfan pharmacokinetics in bone marrow transplant patients: is drug monitoring warranted?

Pharmacokinetics were studied in relation to hepatic side-effects in 20 patients (19 adults aged 18-53 years and one child of 11 years) undergoing BMT after conditioning with 1 mg/kg busulfan (every 6 hours for 16 doses). Busulfan was quantitated in plasma samples at 10 time points within the 6 h dosing interval using HPLC before and after dose numbers 1, 2, 5, 13 and 14. For 13 patients data on all five doses are available; for the remaining seven patients three to four doses were studied. Mean maximum concentrations were 1512 ng/ml; mean trough levels for second and subsequent doses were 615 ng/ml. Maxima (Cmax) tended to be lower and times of maxima (Tmax) were later when busulfan was taken with a meal. Correlation of the area under the concentration versus time curve (AUC0-6h) between different doses was low within patients. In several patients problems with compartmental fitting of concentration data were observed mainly caused by the short dosing interval, which made estimates of T1/2 and model derived AUCs unstable. Three patients experienced hepatic veno-occlusive disease; kinetic parameters were not helpful in describing a particulate risk constellation for this subgroup. In our experience, the role of drug monitoring in this setting needs to be defined more clearly.

Adolescent↗
PubMed1991

Isolation and structure elucidation of urinary metabolites of mitoxantrone.

Three 13C-labeled 1,4-dihydroxy-5,8-bis(2-[(2-hydroxyethyl)amino]-ethyl)amino-9,10- anthracenedione dihydrochloride (mitoxantrone) isotopomers were synthesized to prove the proposed chemical structures of human urinary metabolites by means of nuclear magnetic resonance spectroscopy. After application of labeled mitoxantrone to an anesthetized pig, urine was collected by way of a vesicourethral catheter. The urinary metabolites were isolated by liquid chromatography using a new procedure developed for extraction of mitoxantrone metabolites. Structural elucidation by nuclear magnetic resonance spectroscopy and tandem mass spectrometry confirmed the proposed mono- and dicarboxylic acid structures of the metabolites. High-performance liquid chromatography of native pig urine showed an additional metabolite detected by its UV-visible absorption. The new metabolite was identified as a glucuronic acid conjugate of mitoxantrone by means of nuclear magnetic resonance spectroscopy and tandem mass spectrometry. Incubation with beta-glucuronidase under high-performance liquid chromatography control revealed mitoxantrone as the sole product. Quantitative high-performance liquid chromatography analyses showed that the new urinary metabolite represents the main biotransformational pathway of mitoxantrone in pigs, indicating significant interspecies variation in mitoxantrone biotransformation. Expressed in equivalents of mitoxantrone, the new metabolite amounts to 25% and 31%, respectively, of urinary excreted drug-related material. Extraction of patient urine using the same procedure led to the isolation of pure metabolite B. Tandem mass spectrometric data delivered definitive evidence for the structure of metabolite B as monocarboxylic acid of mitoxantrone.

Animals↗
PubMed1991

Facilitation and suppression of wrist and digit muscles from single rubromotoneuronal cells in the awake monkey.

1. The output effects of 214 cells in the magnocellular red nuclei of two rhesus monkeys (Macaca mulatta) were tested with spike-triggered averaging of electromyogram (EMG) activity from six forearm extensor and six flexor muscles. The monkeys performed an alternating wrist movement task (auxotonic paradigm) or generated wrist torque trajectories alternating between flexion and extension (isometric paradigm). 2. Sixty-five cells (30%) were identified as rubromotoneuronal (RM) cells on the basis of their postpike effects on forearm flexor and extensor muscles. Three major types of RM cell output organization were identified: 1) pure facilitation (28 cells), 2) reciprocal (18 cells), and 3) cofacilitation (16 cells). 3. RM cell output showed a strong preference for facilitation of extensor forearm muscles. This preference was reflected in the fact that 69% (43 of 62) of RM cells facilitated extensors exclusively or most strongly; 27% facilitated flexors exclusively or most strongly; and 5% facilitated flexors and extensors equally. Postspike facilitation (PSpF) was observed in 45% of the extensor muscles and 20% of the flexors tested. In contrast, postpike suppression (PSpS) was observed in 3% of the extensors and 7% of the flexors. 4. The mean number of extensors facilitated per RM cell was 3.1 (53% of tested) compared with 2.8 (51% of tested) flexors facilitated per cell. The extensor and flexor PSpS muscle field sizes were both 2.0 (35% of extensors and 36% of flexors tested). The mean number of muscles facilitated by cofacilitation cells was 5.8 (48%) per cell. No clear preference was found for facilitation of particular combinations of synergist muscles. 5. PSpF magnitude was assessed by measuring both the percent change of facilitation or suppression from baseline and the signal-to-noise ratio of effects. The overall average magnitudes of RM PSpF and PSpS were 4.1 +/- 2.0 and 4.0 +/- 2.3% change from baseline, respectively. The average magnitude of PSpF in flexors was not significantly different from that of extensors; neither was there a difference in the average magnitude of PSpS in flexors and extensors. 6. The mean onset latency of RM cell PSpS was greater than PSpF (9.2 +/- 3.0 vs. 5.7 +/- 1.8 ms; P less than or equal to 0.05). This can be attributed to an underlying minimal disynaptic linkage to motoneurons for suppression effects, whereas most PSpFs are probably mediated by underlying monosynaptic connections. The mean onset latency of flexor PSpFs was greater than that of extensors (6.4 +/- 2.3 vs. 5.4 +/- 1.5 ms; P less than or equal to 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1991

Effects on wrist and digit muscle activity from microstimuli applied at the sites of rubromotoneuronal cells in primates.

1. The purpose of this study was to use the techniques of spike- and stimulus-triggered averaging (SpTA and StTA, respectively) to examine the output organization of individual rubromotoneuronal (RM) cells in relation to clusters of neighboring cells. SpTA of electromyographic (EMG) activity in awake monkeys reveals the target muscles of an individual recorded neuron, whereas StTA reveals the target muscles of the neuronal aggregate activated by the stimulus. 2. Three questions were of particular interest. First, does the pattern of poststimulus facilitation (PStF) across forearm muscles match the pattern of postspike facilitation (PSpF)? Second, does the output of RM cell aggregates tested with StTA favor forearm extensor muscles, as reported for individual RM cells in the companion paper? Third, how do RM poststimulus effects compare with corticomotoneuronal (CM) poststimulus effects? 3. Microstimuli were applied at the sites of 37 RM cells, identified by SpTA of EMG activity in awake monkeys performing an alternating wrist movement task. 4. The magnitudes of PStF at 5, 10, and 20 microA were, respectively, 4.3, 10.1, and 13.7 times greater than PSpF of the same muscles, reflecting activation, by the stimulus, of multiple RM cells. RM cell PStF was weaker than CM PStF. 5. The onset latency of poststimulus suppression (PStS) exceeded that of PStF. For example, at 20 microA the difference was 2.6 ms, comparable with the difference between PSpF and postspike suppression (PSpS). 6. The patterns of poststimulus effects on forearm flexor and extensor muscles were categorized in the same manner as postspike effects. Three major patterns were observed: 1) pure facilitation, 2) reciprocal suppression, and 3) cofacilitation of extensors and flexors. 7. The profile of PStF across synergist muscles was broadly similar to that of PSpF. At 83% of sites, the muscle with the greatest PSpF was also the muscle with the greatest PStF. At 30% of sites (11 of 37), the set of muscles with PStF (muscle field) exactly matched the set with PSpF. Overall, the level of discrepancies in SpTAs versus StTAs computed at the same RM cell sites was 27%. Most of these discrepancies could be attributed to muscles with the weakest effects. 8. The fact that the poststimulus muscle fields at many RM cell sites matched the postspike fields at the same sites, even though the poststimulus effects were greater in magnitude and were mediated by more cells, suggests clustering of RM cells with similar target motoneuron pools.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1989

Control of forelimb muscle activity by populations of corticomotoneuronal and rubromotoneuronal cells.

We review and synthesize evidence on the activity of corticomotoneuronal (CM) and rubromotoneuronal (RM) cells and single motor units in forearm muscles in monkeys performing alternating wrist movements. The CM and RM cells were identified by post-spike facilitation of rectified forelimb EMG activity. RM cells facilitated more muscles per cell (mean: 3.0 of 6 synergist muscles) than CM cells (2.4/6). Both groups had "reciprocal" cells which also suppressed antagonists of their facilitated target muscles. Unlike CM cells, some RM cells cofacilitated flexor and extensor muscles (5.8 or 12 muscles). During performance of a standard ramp-and-hold force tracking task the firing patterns of CM and RM cells, as well as single motor units, fell into distinct response types. Each population had phasic-tonic and tonic cells. Unique to the CM population were cells whose discharge increased during the static hold period; unique to RM cells were bidirectionally responsive and unmodulated neurons. Many motor units showed decrementing discharge. To estimate the ensemble activities of these populations the response histograms of different cells were summed (with force ramps aligned) in proportion to the relative frequency of each cell type. The population response histogram of CM cells was phasic-tonic, consistent with the predominant response type. The population response of RM cells was also phasic-tonic, but showed a shallower phasic modulation relative to discharge that was sustained during both directions of movement. The population histogram of motor units of a muscle was proportional to the average of rectified multiunit EMG, and typically exhibited decrementing activity during the static hold. The effects of excitatory postsynaptic potentials (EPSPs) on firing probability of motoneurons previously documented in intracellular studies are combined with the mean firing rates in the population histograms and the known amplitudes of CM-EPSPs and RM-EPSPs to infer the relative contributions of the supraspinal cells to tonic discharge of active motoneurons. This analysis suggests that for intermediate levels of force, the CM cells would increment motoneuron discharge by about 9 impulses/second (i.p.s.) and RM cells by about 2.4 i.p.s. The analysis also reveals differences in the population activity of CM and RM cells compared to their target motoneurons, which may be due to other input cells and to recruitment properties of motoneurons.

Animals↗
PubMed1985

Implementation of a movement paradigm using the Commodore 64 microcomputer.

Implementation of an alternating movement paradigm for monkeys was achieved using an inexpensive but versatile microcomputer, the Commodore 64. During task performance, the computer monitors one of three user selectable input signals (e.g. joint position) and continuously displays this signal as a moving cursor on a video monitor along with a user positioned target box. Other user defined parameters include in-target holding time and reinforcement ratio. The system also provides two sound cues to signal entry of the cursor into the target box and successful completion of a trial. Extensive use is made of the computer's intrinsic hardware features for implementation of movement paradigm functions. Use of external components is limited to digitizing and interface hardware. A two part software package consisting of a BASIC and a machine language program performs all task and hardware related functions. Acquisition and display of analog input signals, display of target positions, and delivery of auditory cues and applesauce rewards are all controlled by the machine language program. All user defined parameters are specified from the BASIC menu program. The specific programs described in this paper should be applicable to the control of tasks requiring alternation of a behavioral parameter between two target zones.

Animals↗
PubMed1984

Soman induced changes in brain regional glucose use.

Soman, a potent central acetylcholine esterase inhibitor, has a greater impact on brain regional glucose use than other organophosphates, such as diisopropylfluorophosphate (DFP) or phospholinium iodide. At near-lethal doses soman induced explosive persistent seizures that were associated with a greater than fourfold increase of glucose use in many brain structures. Single near-lethal doses of soman lead to conspicuous neuronal damage and a marked reduction in brain activity, 1 to 3 days after exposure. When soman (2 X LD50) was given to TAB (an antidotal mixture of trimedoxime, atropine, and benactyzine ) pretreated rats, there was a greater than twofold reduction of glucose use in almost every brain region. We suggest that soman seizures are mediated via activation of muscarinic receptors; also, the substantia nigra has a key role in the initiation/propagation of seizures. Soman has in addition, a depressive effect on some brain components which appears not to involve muscarinic receptors. We suggest that the conspicuous pathology that follows a single, near-lethal dose of soman results from a depletion of energy flow along with an influx of Ca2+ which sets into motion a cascade of destructive reactions, such as activation of proteases.

Animals↗
PubMed1982

Superior colliculus activation by retinal nicotinic ganglion cells: a 2-deoxyglucose study.

Systemic injection of the acetylcholinesterase inhibitor, di-isopropylfluorophosphate, in rats causes a marked increase in glucose use in the superficial layers of the superior colliculus. This activation of the superior colliculus is largely a retinal effect. Furthermore, since this response can be blocked by intraocular as well as systemic injections of mecamylamine, it is postulated that retinal nicotinic receptors are involved.

Animals↗
PubMedDate not supplied

Encoding of motor parameters by corticomotoneuronal (CM) and rubromotoneuronal (RM) cells producing postspike facilitation of forelimb muscles in the behaving monkey.

This paper compares the properties of corticomotoneuronal (CM) and rubromotoneuronal (RM) cells identified by postspike facilitation (PSF) of rectified EMG activity in the awake monkey. The postspike effects of CM and RM cells in flexors and extensors of the wrist and fingers have been determined, as have the discharge properties of these cells in relation to alternating ramp-and-hold wrist movements. The characteristics of postspike facilitation and postspike suppression (PSS) were similar for RM and CM cells. The magnitude of RM-PSF was weaker than CM-PSF and RM cells showed a stronger preference for facilitation of extensor muscles than CM cells. As with CM cells, the onset of discharge in RM cells preceded the onset of EMG activity in their target muscles. Tonic discharge related to static torque was more prominent in CM cells, whereas phasic discharge was more prominent in RM cells; however, many RM cells showed some tonic activity weakly related to static torque. We conclude that CM and RM cells share many common features; however, RM cells are concerned primarily with the dynamics of muscle contraction.

Animals↗
PubMedDate not supplied

Evidence for oxidative activation of mitoxantrone in human, pig, and rat.

A new metabolite of mitoxantrone in human, rat, and pig urine has been discovered by means of HPLC. The metabolite has been isolated by preparative HPLC from patient urine and is characterized by tandem mass spectrometry and UV-visible spectroscopy as 8,11-dihydroxy-4-(2-hydroxyethyl)-6-[[2-[(2-hydroxyethyl)amino]ethyl] amino]-1,2,3,4,7,12-hexahydronaphtho-[2,3-f]-chinoxaline-7,1 2-dione. Final structural proof has been obtained by independent synthesis. The new metabolite is a product of the enzymatic oxidation of the phenylenediamine substructure of mitoxantrone. An important biological consequence of the oxidative biotransformation is the possibility of covalent binding to intracellular targets via a highly electrophilic intermediate. Thus, alkylation may be an important mode of action of mitoxantrone. Incubation of mitoxantrone with horseradish peroxidase/hydrogen peroxide in the presence of glutathione led to the formation of two glutathione conjugates of mitoxantrone. Their structures have been elucidated by combination of IonSpray (Sciex, Canada) ionization and tandem mass spectrometry. Radioactive mitoxantrone, synthesized from sodium [14C]cyanide, was used to determine interspecies variations between human and rat. The collected rat urine was analyzed by HPLC using a radioactivity monitoring detector and revealed significant differences in the biotransformation of mitoxantrone in rat compared to human. The main metabolites thus far described in human urine are not observed in rat urine.

Animals↗