Biomedical subjects

M Kiefer

Publications and source records attributed to M Kiefer.

At least 37 records · Page 2Linked to original sources

PubMed2001

The interaction of bovine adrenodoxin with CYP11A1 (cytochrome P450scc) and CYP11B1 (cytochrome P45011beta ). Acceleration of reduction and substrate conversion by site-directed mutagenesis of adrenodoxin.

The kinetics of protein-protein interaction and heme reduction between adrenodoxin wild type as well as eight mutants and the cytochromes P450 CYP11A1 and CYP11B1 was studied in detail. Rate constants for the formation of the reduced CYP11A1.CO and CYP11B1.CO complexes by wild type adrenodoxin, the adrenodoxin mutants Adx-(4-108), Adx-(4-114), T54S, T54A, and S112W, and the double mutants Y82F/S112W, Y82L/S112W, and Y82S/S112W (the last four mutants are Delta113-128) are presented. The rate constants observed differ by a factor of up to 10 among the respective adrenodoxin mutants for CYP11A1 but not for CYP11B1. According to their apparent rate constants for CYP11A1, the adrenodoxin mutants can be grouped into a slow (wild type, T54A, and T54S) and a fast group (all the other mutants). The adrenodoxin mutants forming the most stable complexes with CYP11A1 show the fastest rates of reduction and the highest rate constants for cholesterol to pregnenolone conversion. This strong correlation suggests that C-terminal truncation of adrenodoxin in combination with the introduction of a C-terminal tryptophan residue enables a modified protein-protein interaction rendering the system almost as effective as the bacterial putidaredoxin/CYP101 system. Such a variation of the adrenodoxin structure resulted in a mutant protein (S112W) showing a 100-fold increased efficiency in conversion of cholesterol to pregnenolone.

Adrenodoxin↗

PubMed2001

Thermal sterilization of heat-sensitive products using high-temperature short-time sterilization.

High-temperature short-time (HTST) sterilization with a continuous-flow sterilizer, developed for this study, was evaluated. The evaluation was performed with respect to (a) the chemical degradation of two heat-sensitive drugs in HTST range (140-160 degrees C) and (b) the microbiological effect of HTST sterilization. Degradation kinetics of two heat-sensitive drugs showed that a high peak temperature sterilization process resulted in less chemical degradation for the same microbiological effect than a low peak temperature process. Both drugs investigated could be sterilized with acceptable degradation at HTST conditions. For the evaluation of the microbiological effect, Bacillus stearothermophilus ATCC 7953 spores were used as indicator bacteria. Indicator spore kinetics (D(T), z value, k, and E(a)), were determined in the HTST range. A comparison between the Bigelow model (z value concept) and the Arrhenius model, used to describe the temperature coefficient of the microbial inactivation, demonstrated that the Bigelow model is more accurate in prediction of D(T) values in the HTST range. The temperature coefficient decreased with increasing temperature. The influence of Ca(2+) ions and pH value on the heat resistance of the indicator spores, which is known under typical sterilization conditions, did not change under HTST conditions.

Geobacillus stearothermophilus↗

PubMed2001

Perceptual and semantic sources of category-specific effects: event-related potentials during picture and word categorization.

In two experiments the effect of object category on event-related potentials (ERPs) was assessed while subjects performed superordinate categorizations with pictures and words referring to objects from natural (e.g., animal) and artifactual (e.g., tool) categories. First, a category probe was shown that was presented as name in Experiment 1 and as picture in Experiment 2. Thereafter, the target stimulus was displayed. In both experiments, analyses of the ERPs to the targets revealed effects of category at about 160 msec after target onset in the pictorial modality, which can be attributed to category-specific differences in perceptual processing. Later, between about 300-500 msec, natural and artifactual categories elicited similar ERP effects across target and category modalities. These findings suggest that perceptual as well as semantic sources contribute to category-specific effects. They support the view that semantic knowledge associated with different categories is represented in multiple subsystems that are similarly accessed by pictures and words.

Adult↗

PubMed2001

[Pressure-dependent outflow resistance in cerebrospinal fluid dynamics: evaluation a calculation model for diagnosis of normal pressure hydrocephalus in an animal experiment with H-Tx rats].

The internationally accepted methods of calculating cerebrospinal fluid dynamics proceed from the assumption of a pressure-independent resistance to CSF outflow. Our new model focusses on the pressure-dependency of this resistance. In it, we monitor the entire pressure course over time, p(t) during and after infusion. A comparison of the pressure rise, On(p), during infusion, and the decrease, Off(p), to the same pressure level, permits the creation of all the formulas for C(p) and R(p). The simultaneous measurement of resistance and compliance during a single intervention allows us to minimize patient exertion. In contrast to the classical methods, it is not necessary for the ICP to reach a plateau. Our mathematical model differs from the static examination model by describing a pressure-dependent slope of the function for the resistance. This has been demonstrated in a study using H-Tx rats. In this way, we are able to take the non-linearity of the CSF resorption into consideration.

Animals↗

PubMed2001

[The importance of hydrostatic valves in the treatment of adult chronic hydrocephalus].

BACKGROUND AND OBJECTIVE: Earlier designs of hydrocephalus valves, focusing on a reduction of the problem of overdrainage, were proved to be insufficiently. Since the middle of the 90's hydrostatic valve, constructions are available, which should solve this problem. The objective of the study was to evaluate, whether these gravitational shunts would indeed reduce the percentage of overdrainage in shunted patients. METHODS: In 51 patients clinical evaluation and MRI was performed pre- and in regular intervals postoperatively. 21 received a Dual-Switch valve, 14 a programmable Codman Hakim plus Shuntassistent and 6 a combination of a conventional valve with Shuntassistent. RESULTS: Over 90% of the patients had clear and persisting (over 24 months) clinical benefit from shunting. Only one suffered for some days after shunt implantation from headache in the upright position. No other clinical or imaging hints were given on an overdrainage. The ventricular size was unchanged in patients with a normal pressure hydrocephalus after shunting. In patients with occlusive hydrocephalus only a few showed a significant reduction of the ventricular wide. CONCLUSION: Gravitational shunts seem to decrease the overdrainage linked to the treatment of chronic hydrocephalus in shunted patients, therefore its use should be standardised when treatment chronic hydrocephalus.

Adult↗

PubMed2000

Structural basis for substrate specificity differences of horse liver alcohol dehydrogenase isozymes.

A structure determination in combination with a kinetic study of the steroid converting isozyme of horse liver alcohol dehydrogenase, SS-ADH, is presented. Kinetic parameters for the substrates, 5beta-androstane-3beta,17beta-ol, 5beta-androstane-17beta-ol-3-one, ethanol, and various secondary alcohols and the corresponding ketones are compared for the SS- and EE-isozymes which differ by nine amino acid substitutions and one deletion. Differences in substrate specificity and stereoselectivity are explained on the basis of individual kinetic rate constants for the underlying ordered bi-bi mechanism. SS-ADH was crystallized in complex with 3alpha,7alpha,12alpha-trihydroxy-5beta-cholan -24-acid (cholic acid) and NAD(+), but microspectrophotometric analysis of single crystals proved it to be a mixed complex containing 60-70% NAD(+) and 30-40% NADH. The crystals belong to the space group P2(1) with cell dimensions a = 55.0 A, b = 73.2 A, c = 92.5 A, and beta = 102.5 degrees. A 98% complete data set to 1.54-A resolution was collected at 100 K using synchrotron radiation. The structure was solved by the molecular replacement method utilizing EE-ADH as the search model. The major structural difference between the isozymes is a widening of the substrate channel. The largest shifts in C(alpha) carbon positions (about 5 A) are observed in the loop region, in which a deletion of Asp115 is found in the SS isozyme. SS-ADH easily accommodates cholic acid, whereas steroid substrates of similar bulkiness would not fit into the EE-ADH substrate site. In the ternary complex with NAD(+)/NADH, we find that the carboxyl group of cholic acid ligates to the active site zinc ion, which probably contributes to the strong binding in the ternary NAD(+) complex.

Alcohol Dehydrogenase↗

PubMed2000

Time course of conscious and unconscious semantic brain activation.

Event-related brain potentials (ERPs) were used to investigate the neurophysiological correlates of conscious and unconscious semantic processing. Masked words, which do not enter consciousness, as well as visible words, were shown to modulate the N400 ERP potential to subsequently presented, meaningfully related target words. The N400 is known as an electrophysiological index of semantic processing. However, for unconsciously perceived words semantic brain activation decays fast within 200 ms, but increases with time for consciously perceived words. Thus, conscious and unconscious semantic activation involves similar brain areas, but both forms of processes exhibit distinct, qualitatively different time courses.

Adult↗

PubMed2000

Transient kinetics of ligand binding and role of the C-terminus in the dUTPase from equine infectious anemia virus.

Transient kinetics of the equine infectious anemia virus deoxyuridine 5'-triphosphate nucleotide hydrolase were characterized by monitoring the fluorescence of the protein. Rate constants for the association and dissociation of substrate and inhibitors were determined and found to be consistent with a one-step mechanism for substrate binding. A C-terminal part of the enzyme presumed to be flexible was removed by limited trypsinolysis. As a result, the activity of the dUTPase was completely quenched, but the rate constants and fluorescent signal of the truncated enzyme were affected only to a minor degree. We conclude that the flexible C-terminus is not a prerequisite for substrate binding, but indispensable for catalysis.

Animals↗

PubMed2000

Kinetic and spectroscopic characterization of native and metal-substituted beta-lactamase from Aeromonas hydrophila AE036.

Two metal ion binding sites are conserved in metallo-beta-lactamase from Aeromonas hydrophila. The ligands of a first zinc ion bound with picomolar dissociation constant were identified by EXAFS spectroscopy as one Cys, two His and one additional N/O donor. Sulfur-to-metal charge transfer bands are observed for all mono- and di-metal species substituted with Cu(II) or Co(II) due to ligation of the single conserved cysteine residue. Binding of a second metal ion results in non-competitive inhibition which might be explained by an alternative kinetic mechanism. A possible partition of metal ions between the two binding sites is discussed.

Aeromonas hydrophila↗

PubMed2000

Executive control is disturbed in schizophrenia: evidence from event-related potentials in a Go/NoGo task.

BACKGROUND: Schizophrenic patients suffer from cognitive and attentional deficits, particularly from failure of executive control functions. METHODS: This study investigated the cortical organization of executive control in schizophrenic patients and healthy control subjects using event-related potentials (ERPs). Event-related potentials were collected while subjects performed an auditory Go/NoGo task that required response inhibition. To exclude stimulus discriminability and early stimulus processing to confound results, stimuli were adjusted to the subject's individual discrimination ability and were presented in a simple and a difficult version. RESULTS: Schizophrenic patients performed similar to control subjects in the Go condition but worse than control subjects in the NoGo condition that required response inhibition. Event-related potentials revealed the neurophysiological substrate of this dysfunction. In the Go conditions, both healthy control subjects and schizophrenic patients showed the same voltage pattern. In the NoGo condition, control subjects and patients showed similar cortical activation only during early processing (N2 time window). However, in later stages of processing (P3 time window), healthy subjects showed left lateralization of ERPs over frontal areas while schizophrenic patients did not. CONCLUSIONS: We conclude that schizophrenic patients exhibit deficient processing in a neuronal network, including left frontal areas, that is involved in later stages of executive control function.

Adult↗

PubMed2000

Health complaints related to pesticide stored at a public health clinic.

Employees at a health center in Georgia were concerned that symptoms experienced by some employees were related to pesticide exposure at the center. Malathion and DDT, used for mosquito control from 1969 to 1981, had been stored and handled at the center's first floor. We surveyed 117 (91%) of 129 employees to determine whether reported symptoms were associated with pesticide exposure. We performed environmental sampling for pesticides. We analyzed serum samples for 17 chlorinated pesticides, and urine samples for malathion. We found that 37% of the participants had reported a diagnosis of sinusitis and 24% of bronchitis since working at the health center. Frequently reported symptoms were eye irritation (44%) and headache (68%). DDT and malathion were found at levels of 2.4 and 11%, respectively, in bulk samples from the loading dock of the building. Multivariate analysis of responses to the questionnaire showed that the perception of odors, inadequate air flow, and length of employment were significantly associated with the employees' health complaints. Pesticide concentrations in employees' serum and urine samples were not associated with any health complaint. The health complaints reported by the employees at the health center were precipitated by both environmental and psychological factors. The epidemiology and laboratory components of this study highlight the importance of obtaining biological measurements in episodes of perceived environmental exposure.

Adult↗

PubMed2000

The dynamic infusion test in rats.

Although the hydrocephalic H-Tx rat is a widely used model, data on the cerebrospinal fluid (CSF) dynamics in hydrocephalic rats are rare or - as the pressure volume index (PVI) - not available. We used hydrocephalic and nonhydrocephalic H-Tx rats, a stock with a high percentage of inherited hydrocephalus, for the evaluation of such data. In addition, a new, simple mathematical algorithm ("dynamic infusion test"), which has not formerly been used in animal experiments, was used as a pathophysiological model of CSF dynamics. Compared with classical methods for evaluation of these data, the dynamic infusion test gives a deeper insight into the relation between ICP and CSF dynamics. It was found that the resistance to outflow (ROF) in hydrocephalic rats was at least twice that in nonhydrocephalic rats. The PVI measured was similar in hydrocephalic and nonhydrocephalic animals, but clearly higher than the values reported in the literature. This may be attributable to the fact that the classically used bolus test, in contrast to the "dynamic infusion test", is representative only for the CSF compartment which is directly exposed to the bolus application.

Algorithms↗

PubMed2000

[Significance of hydrostatic valves in therapy of chronic hydrocephalus].

Earlier design changes in hydrocephalus valves focusing on reducing overdrainage failed. Since the middle of the 1990s, hydrostatic valve constructions have been available which are claimed to solve this problem. The objective of this study was to evaluate the efficiency of these constructions. Clinical status and ventricular size were evaluated in 45 patients with chronic hydrocephalus before and 1, 6, and 26 weeks after operation. In 35 of these, a Miethke dual-switch valve was implanted (treatment group 1), and the others received a combination of a programmable Codman-Hakim valve and a Miethke shunt assistant as the hydrostatic element of the configuration (treatment group 2). A third group (n = 6) had already been shunted but suffered from overdrainage symptoms which could not be overcome by conservative measures. In these cases, the only operative treatment was the implantation of a Miethke shunt assistant adjunctively to the existing valve. In groups 1 and 2, there was a significant permanent improvement in the clinical state in nearly 80% of cases and a moderate permanent improvement in about 10%. Mild clinical and radiological signs of overdrainage occurred in three patients during the first postoperative week but resolved without further operative measures within the next 5 weeks. Typically, the ventricular width was not or only marginally reduced in these 45 patients. In the patients treated for symptoms of overdrainage (group 3), complaints resolved within the first week after implantation of the shunt assistant. The study indicates that gravitational shunts may be very effective in preventing overdrainage in chronic hydrocephalus, and therefore these constructions could represent the gold standard in the treatment of chronic hydrocephalus.

Adult↗

PubMed1999

Electrophysiological correlates of direct versus indirect semantic priming in normal volunteers.

The N400 component of event related potentials (ERP) was studied in 27 right-handed healthy subjects in a speeded lexical decision task. The semantic distance between primes (always words) and targets (50% words, 50% non-words) was systematically varied. Prime-target relations included directly related words (hen-egg), indirectly related words (lemon-sweet), and non-related words (sofa-wing). ERPs were recorded from 20 scalp electrodes positioned according to the international 10-20 system. The N400 reflected semantic distance with the most negative and latest N400 peak in the non-related condition, the least negative and earliest N400 peak in the direct condition and the peak in the indirect condition in-between. Hence, N400 priming effect in the indirect condition was obtained in the absence of strong sentential constraints and even though the mediating word was physically not present.

Adult↗

PubMed1999

Tracking the time course of object categorization using event-related potentials.

Object categorization processes were investigated by measuring event-related potentials while subjects categorized objects at the superordinate (e.g. animal), basic (e.g. dog) and subordinate (e.g. beagle) levels of abstraction. An enhanced negative deflection (N1) was found at posterior recording sites for subordinate level categorizations compared with basic level categorizations and was interpreted as a marker of increased visual analysis. In contrast, superordinate level categorizations produced a larger frontal negativity relative to basic level categorizations and were interpreted as an indicator of increased semantic processing. These results suggest a neurophysiological basis for the separate cognitive processes responsible for subordinate and superordinate object categorizations.

Adult↗

PubMed1999

Identification of somatosensory pathways by focal-cooling-induced changes of somatosensory evoked potentials and EEG-activity--an experimental study.

OBJECT: Function-preserving neurosurgery requires methods to identify functionally important CNS-areas intraoperatively. We investigated whether a combination of focal cerebro-cortical cooling and monitoring of somatosensory evoked potentials (SEP) is suited for this task, i.e. whether it is able to outline structures belonging to the somatosensory pathway. METHODS: In 17 Wistar rats the somatosensory cortex was focally cooled by 20 degrees C below the initial tissue temperature for periods of five minutes. A cryoprobe with a tip diameter of 3 mm was used and tissue temperatures were measured below and at different distances to the cryoprobe. Tibial nerve evoked SEPs and EEG-spectra were recorded continuously. RESULTS: During cortical cooling the SEP-responses showed a marked delay and amplitude increase of the cortically generated components P13 and N18 and a small latency increase of the subcortically generated wave III. EEG-spectra were depressed mainly in the low frequency range. All cooling effects were reversible and in light- as well as electron-microscopic examinations no tissue damage was found. CONCLUSIONS: Focal cooling of the cortex induces easily recognizable and reversible changes of the bio-electrical activity without causing any histological damage. Therefore the method seems suitable for identifying eloquent areas. It can be expected that clinical application of the cooling technique in combination with intraoperative electrophysiological monitoring will be helpful to further lower the risk of neurosurgical operations. We propose that cooling mainly interferes with the synaptic transmission within the somatosensory cortex, because the observed amplitude increase can be explained by cold-induced depression of inhibitory cortical activity (disinhibition).

Animals↗

PubMed1998

Right hemisphere activation during indirect semantic priming: evidence from event-related potentials.

Healthy subjects performed a lexical decision task in a semantic priming paradigm while event-related potentials (ERPs) were recorded from 64 channels. Semantic distance between prime and target was varied by including directly, indirectly, and nonrelated word pairs. At centro-parietal electrodes an N400 to nonrelated pairs was elicited bilaterally which was sensitive only to direct, but not to indirect semantic priming. These N400 priming effects were mirrored by the RT data. At inferior fronto-temporal sites directly related words showed ERP priming effects over both hemispheres. However, indirectly related words only elicited ERP priming effects over the right hemisphere. These results support the hypothesis that the right hemisphere semantic system is involved in processing of remote semantic information.

Adult↗

PubMed1998

The time course of brain activations during response inhibition: evidence from event-related potentials in a go/no go task.

The cortical organization of executive control was investigated using event-related potentials (ERPs). ERPs were collected while subjects performed a go/no go task that required response inhibition. First, around 260 ms after stimulus onset, an effect of response inhibition on ERPs was observed over inferior prefrontal areas. Generators in these regions were confirmed by source analysis. Later, between 300-600 ms after stimulus onset, a left lateralized fronto-central ERP effect was found which differed in topography from a non-specific effect of task difficulty. Source analysis indicated that generators in anterior cingulate and left premotor areas also contributed to this effect. Orchestrated activation of prefrontal areas and the anterior cingulate subserves executive function whereas relatively late activity of the left premotor cortex is involved in motor control.

Adult↗