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Biomedical subjects

K H Schmidt

Publications and source records attributed to K H Schmidt.

At least 73 records · Page 4Linked to original sources

Viscosity measurements of suspensions of hemoglobin-containing liposomes of varying lipid composition.

The viscosity of suspensions of hemoglobin-containing liposomes (Hb liposomes) and of liposomes without Hb was determined as a function of the shear rate. Liposomes of varying lipid composition were prepared by detergent dialysis. All liposome suspensions showed non-Newtonian viscous behavior, as did erythrocyte suspensions. The viscosity of suspensions of Hb liposomes with a bilayer consisting of egg yolk lecithin and phosphatidylserine was almost identical to erythrocyte suspension viscosity. When cholesterol was included in the lipid component of the Hb liposomes, the dependence of viscosity on the shear rate decreased (flattening of the curve), but overall suspension viscosity became higher than that of erythrocyte suspensions. Evidence is provided for the lipid concentration at which liposomes can be applied intravenously.

Blood Substitutes↗

Ectopic bone induction by partially purified bone extract alone or attached to biomaterials.

Formation of new bone is suggested to be caused by interaction of a set of chemical factors with mesenchymal target cells. A specific assembly of factors, i.e. chemotaxis-, adhesion-, proliferation- and differentiation-factors, as well as macromolecular structure components, essential for formation of large amounts of ectopic bone was termed "osteopoetin" (OP). In the present study a partially purified osteopoetin containing bovine bone extract (OCBE) was used to induce ectopic bone formation. In order to reduce the amount of OCBE necessary for bone induction, OCBE was seeded onto different commercially available collagens or poly-L-lactic acids (PL). Solid collagens in a sponge-like form were used for the first time to function as an attachment system for an osteoinductive substance. The test substances were implanted into abdominal muscle pouches of male Wistar rats. After 21 days the implants were harvested and evaluated histologically. OCBE resulted in the formation of large ossicles containing hematopoietic bone marrow. The minimal amount of OCBE to elicit ectopic bone formation can be reduced by a factor of 10 when attached to quickly resorbable collagens, but not when attached to slowly resorbable PL. These results suggest that collagens are suitable OCBE attachment systems and useful for clinical application.

Animals↗

Non-leaky vesiculation of large unilamellar vesicles (LUV) induced by plasma high density lipoproteins (HDL): detection by HPLC.

Interaction of large unilamellar phosphatidylcholine vesicles (LUV, 75nm) and plasma high density lipoproteins (HDL) resulted in a non-leaky vesiculation of LUV. This vesiculation was detected by a HPLC-system consisting of a combination of three TSK-gel columns (6000PW, 5000PW, 3000SW). With increasing incubation time liposomal [14C]PC, entrapped [3H]inulin, and apoprotein of HDL origin decreased. The decrease was accompanied by a formation of new particles, consisting of liposomal PC and apoprotein. These particles also enclosed [3H]inulin, reflecting a hydrophilic inner space. The formation of the particles reached a maximum after one day of incubation. Retention time was 21 minutes for LUV, 28 minutes for the new particles, and 36 minutes for HDL. In vesicles with membranes consisting of phosphatidylcholine and 30% cholesterol no interactions were observed.

Carbon Radioisotopes↗

Preparation of hemoglobin-containing liposomes using octyl glucoside and octyltetraoxyethylene.

Hemoglobin (Hb) was encapsulated in phosphatidylcholine vesicles by removal of the detergent n-octyl beta-D-glucoside (OG) or n-octyltetraoxyethylene (C8E4) out of mixed detergent-lipid micelles in Hb solution. Three types of apparatus were used for dialysis. Dialysis buffer flow rates, the surface area of the dialysis membranes, and detergent-lipid interactions determined the rate of dialysis, which influenced liposome size and lamellarity. Slow dialysis led to the formation of multilamellar liposomes, at increased dialysis rates Hb liposomes became smaller and unilamellar. Hb was enclosed at highest concentrations in larger liposomes, which included the negatively charged lipid phosphatidylserine or phosphatidic acid as a membrane component. Co-encapsulation of the allosteric factor inositol hexaphosphate led to oxygen dissociation curve values almost identical to those of whole blood. The oxygen-release capacity of Hb liposome suspensions in the physiological partial pressure range was comparable to whole blood. Storage of Hb liposomes for 2 months leaves oxygen-carrying characteristics virtually unchanged, with met-Hb levels increasing to only 11% of total Hb. Preparation of Hb liposomes by dialysis of octyl glucoside or C8E4 is a mild and efficient method for encapsulation of Hb. Since these Hb liposomes can be produced in scale-up batch sizes, they are a candidate for use as an oxygen-carrying blood surrogate.

Blood Substitutes↗

A comparison of coronary risk factors in groups of trained and untrained adolescents.

Fifty-five experimental (29 male, 25 female) and 38 control (20 male, 18 female) adolescent subjects participated in this study to investigate the differences in coronary heart disease (CHD) risk factors in groups of trained and untrained adolescents. As expected the trained group (both sexes) had higher maximal aerobic power (VO2max) and lower systolic blood pressure at rest. The level of total cholesterol was the same in both groups, but the levels of high-density lipoprotein and its lipoprotein subfractions apolipoprotein (Apo-A) and Apo-A1 were higher, and low-density lipoprotein, Apo-B and triglycerides were significantly lower in the experimental group. The value of risk factors from the family history was the same in both groups, but the behavioural and physical risk factors such as smoking and percentage of body fat were lower in the trained group. It would appear that the group of adolescents, trained for several years in athletics and swimming, had a more beneficial lipoprotein profile and a lower level of behavioural and physical risk factors than the control group. For methodological reasons it remains an open question whether these profile differences are the consequences of self-selection procedures or the effects of training.

Adolescent↗

[Studies on the effect of mistletoe extract on the chemotactic activity and on chemiluminescence of polymorphonuclear leukocytes in vitro].

In the present study, the chemotactic activity of human polymorphonuclear granulocytes was measured using the tripeptide N-F-Met-Leu-Phe and different concentrations of extracts of Viscum album as chemotactic stimuli. It turned out that the extracts have the capability of stimulating chemotaxis up to 30% of the formylated tripeptide. Incubation of isolated granulocytes for 20 minutes in medium alone prior to assessment of chemotaxis results in a 20% loss of chemotactic activity as compared to the freshly isolated cells. Incubation in a medium containing the mistletoe extract does not result in partial deactivation of chemotaxis. This protective effect of the plant extract can be interpreted on the basis of an interaction with the oxidative metabolism of the granulocytes causing partial self-deactivation of chemotaxis. Interference of the extract of Viscum album with the oxidative metabolism of phagocytes is shown in experiments with luminol enhanced chemiluminescence stimulated by opsonized zymosan. The data show a dose-dependent decline of photon emission in the presence of the extracts. The in vitro data measured in the present study provide additional evidence of an immunomodulatory effect of extracts of Viscum album from which in vivo therapeutic effects can be expected.

Amino Acid Sequence↗

[Quantitative volumetric determinations on MR tomograms in communicating hydrocephalus].

For patients with communicating hydrocephalus the implantation of a cardioventricular shunt is mostly an approved therapy. However, indications are frequently problematic and therefore a shunt operation will not be always successful. The proof of periventricular oedemas in addition to clinical signs seems to be a criterion for an indication for implantation and for the selection of a special valve. With the aid of a newly developed computer programme volume estimation of the ventricular system and the periventricular oedemas has been effected for six patients on T2-weighted MR-tomograms (1.5 tesla). Quotients of the different volumes are correlated with cerebrospinal fluid (CSF) pressure obtained during shunt implantation or lumbar measurement. Distinct indications have been found for a correlation between the volume of the periventricular oedemas and the pressure measurements.

Brain Edema↗

Subjective health status of day and shift-working policemen.

To assess the subjective health status of day- and shift-working police officers, a questionnaire-based study was carried out. All the day-workers had previous shift experience. To control the age factor the total population of 2659 shift-working and 1303 day-working police officers was divided into four ten-year age classes. Factor analysis revealed that all the symptoms included in the questionnaire could be grouped into six factors. The prevalence rates of complaints showed that four of these factors (autonomous symptoms, musculo-skeletal symptoms, disturbance of appetite and indigestion, respiratory infections) were influenced by the main effects of age and shift work. Across all age classes the age-related changes in prevalence rates were strengthened by shift-work. In the other two factors (nervous symptoms, gastro-intestinal symptoms) an additional interaction effect could be observed. While the prevalence rates of the day-workers increased with age, those of the shift-workers decreased in the oldest age class. This drop of prevalence rates may be attributed to the influence of selection processes.

Adult↗

Complete secretion of activable bovine prochymosin by genetically engineered L forms of Proteus mirabilis.

To circumvent problems encountered in the synthesis of active chymosin in a number of bacteria and fungi, a recombinant DNA L-form expression system that directed the complete secretion of fully activable prochymosin into the extracellular culture medium was developed. The expression plasmid constructions involved the in-frame fusion of prochymosin cDNA minus codons 1 to 4 to streptococcal pyrogenic exotoxin type A gene (speA') sequences, including the speA promoter, ribosomal binding site, and signal sequence and five codons of mature SpeA. Secretion of fusion prochymosin enzymatically and immunologically indistinguishable from bovine prochymosin was achieved after transformation of two stable protoplast type L-form strains derived from Proteus mirabilis. The secreted proenzyme was converted by autocatalytic processing to chymosin showing milk-clotting activity. In controlled laboratory fermentation processes, a maximum specific rate of activable prochymosin synthesis of 0.57 x 10(-3)/h was determined from the time courses of biomass dry weight and product formation. Yields as high as 40 +/- 10 micrograms/ml were obtained in the cell-free culture fluid of strain L99 carrying a naturally altered expression plasmid of increased segregational stability. The expression-secretion system described may be generally useful for production of recombinant mammalian proteins synthesized intracellularly as aberrantly folded insoluble aggregates.

Animals↗

Serum selenium levels in patients with malignant melanoma.

The incidence of malignant melanoma of the skin has increased rapidly among white people during the last decade. Although the pathogenesis of malignant melanoma is not clear, epidemiologic data and experimental work indicates that ultraviolet (UV) radiation plays a critical role in tumorigenesis. Recent data implicate peroxidative reactions in UV-carcinogenesis and clearly demonstrate that selenium (Se) confers protection, in part by inducing cellular-free radical scavenging systems and by enhancing peroxide breakdown, thus enhancing the capacity of the cell to cope with oxidant stress. With this in mind, we investigated the Se status of 101 patients with malignant melanoma. Our study revealed significantly lower Se levels in the sera of melanoma patients than of controls. Although patients in all clinical stages had lower Se levels than the controls, patients in stage III (disseminated melanoma) had the lowest levels. Separate analysis of histogenetic subtypes of melanoma revealed that lentigo maligna melanoma (LMM) and superficial spreading melanoma (SSM) had the strongest association with depressed Se serum levels. Our results, showing for the first time that malignant melanoma is associated with low Se concentrations, are consistent with results of epidemiologic studies showing an inverse correlation between serum Se levels and certain cancers.

Adult↗

[Effect of performance and state orientation on achievement and attention indicators in a simple stimulus-response task with and without performance feedback].

In formulating his theory of action control, Kuhl (1983) proposed two motivational states: action orientation and state orientation. People who are action oriented are able to ignore all informations in concrete situations which are irrelevant with regard to the preferred goals. Persons who are state oriented tend to focus sooner on the outlast aspects of a special state. Kuhl introduced a questionnaire HAKEMP to record these tendencies. Further on, there occurs the general assumption that in a particular state of state orientation, people have only a limited processing capacity for given tasks. This hypothesis was tested in an experiment (n = 88) with a stimulus-reaction task. The conditions were varied by the type of the task (fixed or variable stimulus interval) and feedback for the subjects' ability to react. It was expected that for the fixed stimulus interval condition, state-oriented people would be less efficient than action-oriented subjects; with feedback there should be increasing results. The experiment however, shows no significant difference between the performance of state- and action-oriented subjects. These results are discussed in relation to modifications of the HAKEMP questionnaire.

Adolescent↗

[Stress and strain analysis in monitoring functions in a fully automated production plant].

The introduction of new technologies in the automobile industry produces a substantial structural change. This change may lead to a decrease in physical work with a contrary increase in mental and emotional stress at monitoring and controlling tasks. A summarizing analysis of the stress was made by means of the "Arbeitswissenschaftliches Erhebungsverfahren zur Tätigkeitsanalyse (AET)" and for characteristic working conditions the energy expenditure during work was measured. As indicators of strain, the heart rate was registered continuously with portable magnetic tape recorders and additionally the urinary excretion of adrenaline and noradrenaline was analyzed. During and after the implementation of new technologies a shift from physical to mental and/or emotional stress could be observed, but the remaining components of physical load are not to be disregarded and the changing strain patterns have to be analyzed carefully during the transitional stages of the implementation process.

Automation↗

Structural changes in vesicle membranes and mixed micelles of various lipid compositions after binding of different bile salts.

Binding equilibria of common bile salts (BS) and different mixtures of membrane lipids were correlated with BS-induced structural changes of large unilamellar vesicles, with transition of vesicles to mixed micelles (MM), and with successive transformations of MM. At very low BS concentrations, in the outer vesicle monolayer definite BS/lipid aggregates are formed, the size and BS binding strength of which depend on the BS and lipid species involved. At increasing BS concentrations, binding to the membranes is hampered, and above a critical BS content, membrane stress due to asymmetric BS binding leads to formation of transient membrane holes, as shown by inulin release from the vesicles. Independent of the BS and lipid species, membrane solubilization starts at a ratio r = 0.3 of bound BS/lipid. Increasing phosphatidylserine, phosphatidylethanolamine, and cholesterol contents stabilize the lecithin membrane against BS to different degrees and in different ways, whereas the destabilization by sphingomyelin is probably due to the enhancement of the membrane gel-liquid transition temperature. Conjugation of the BS with glycine or taurine has a modulating effect on membrane hole formation, rather than on lipid solubilization. Diphenylhexatriene fluorescence anisotropy indicates a BS-induced drop of the internal membrane order and its restoration during membrane solubilization. At higher concentrations ursodeoxycholate induces additional condensation, whereas the other BS cause internal disorder in the MM. Above ratios r of approximately 8:1, we found a release of BS from these MM and suggest a rodlike structure for them. The results were discussed with respect to BS/membrane interactions during lipid excretion from the liver cell.

Bile Acids and Salts↗

Tripartite streptokinase gene fusion vectors for gram-positive and gram-negative procaryotes.

A specific 1,596 bp HincII fragment ('skc) from the chromosome of Streptococcus equisimilis contains an active streptokinase (SK) gene (skc) lacking, in addition to the expression signals, codons 1 through 39 of wild-type skc but retaining the remainder of the skc coding sequence together with the transcription terminator. Using this fragment as an indicator gene, we constructed two types of vectors which in appropriate hosts resulted in the synthesis of SK fusion proteins after insertional activation of 'skc. The first type are open reading frame (ORF) vectors in which 'skc was inserted into pUC18 out of frame with respect to lacZ', thus conferring an SK-negative phenotype. Any DNA fragments representing ORFs inserted between the lacZ' expression signals and 'skc such that the skc reading frame was restored resulted in the production of tripartite proteins which exhibited SK activity. The second type of vector, which functioned in both gram-positive and gram-negative bacteria, used the streptococcal speA expression and secretion signals in front of the ORF to activate 'skc insertionally. Using a large fragment from the chymosin gene as the target sequence, the usefulness of these vectors for studying foreign gene expression in streptococci as well as Escherichia coli was demonstrated.

Amino Acid Sequence↗

Work at sea: a study of sleep, and of circadian rhythms in physiological and psychological functions, in watchkeepers on merchant vessels. II. Sleep duration, and subjective ratings of sleep quality.

Sleep length and sleep quality scores were collected on board ships over periods of up to two weeks from 38 watchkeepers working a '4-on/8-off routine' and 29 dayworkers. All watchkeepers exhibited fragmented sleeping patterns, which indicated a lack of adaptation of the sleep/wakefulness cycle to the hours of work. There were only slight differences in total sleep length between watchkeepers and dayworkers, however, both groups did not obtain an adequate amount of sleep. Within the watchkeeping crews the 3rd Officers had by far the shortest sleep length. Concerning sleep quality, daytime sleep was generally given the lowest ratings, whereas sleep starting before midnight was on average evaluated as the best, both by watchkeepers and dayworkers. Watchkeeping personnel do not normally have any "days off" during a voyage so that missed sleep might even amount to a sleep deficit. A solution for this problem could perhaps be a new, stabilized system that allows a single uninterrupted sleep, which is required for full recuperation, to be taken each day.

Circadian Rhythm↗

Localization and characterization of fibronectin-binding to group A streptococci. An electron microscopic study using protein-gold-complexes.

The location and nature of the binding sites for fibronectin (Fn) and its N-terminal 29 K fragment (FnF) on group A streptococci were studied by electron microscopy using these proteins labelled with colloidal gold. The investigated strains exhibited a different labelling intensity as well as a different labelling pattern varying from a strong regular distribution to a weak focal binding. Binding of Fn and FnF was inhibited by itself as well as by lipoteichoic acid (LTA), anti-LTA and concanavalin A. Simultaneous labelling of the bacteria with marker complexes of FnF, human serum albumin and fibrinogen revealed separate receptor sites for each protein. Our results confirmed LTA to be mainly responsible for the binding of Fn on group A streptococci.

Binding Sites↗

Changes in the pattern of microsomal fatty acids in rat liver after thermal injury and therapeutic intervention.

Despite substantial progress in handling the acute phase after thermal injury, severely burned patients still succumb to systemic sepsis as a consequence of a compromised defence system. It is likely that autotoxic mechanisms play an important role in the aetiology of the impaired host defence. One of the primary target systems of autotoxic cell damage is the liver. In the present work oxidative alterations in the microsomal compartment of liver cells have been investigated. It was found that thermal burns are associated with extensive oxidation of polyunsaturated fatty acids which can be antagonized by antioxidants such as silibinin.

Animals↗

L-ascorbic acid modulates 5-lipoxygenase activity in human polymorphonuclear leukocytes.

Leukotriene B4 (LTB4), a metabolite of the arachidonic acid pathway mediated by 5-lipoxygenase, is released by stimulated polymorphonuclear neutrophils (PMNL) and has been postulated to be an important mediator of the inflammatory response. Extracellular L-ascorbate at concentrations in the range of 0.5-5.0 mM effectively inhibits the biosynthesis of LTB4 and 5-hydroxyeicosatetraenoic acid (5-HETE) stimulated by the calcium ionophore A 23187. The ionophore-activated LTB4 production is reduced after incubation of PMNL with opsonized zymosan as a phagocytic stimulus. Extracellular L-ascorbate at concentrations above 0.1 mM reverses the zymosan-induced deactivation of 5-lipoxygenase, resulting in significantly higher LTB4 and 5-HETE yields. The inhibitory effect of zymosan preincubation on LTB4 production is independent of calcium or free arachidonic acid in the incubation medium. Interaction of L-ascorbate with the catabolism of LTB4 was excluded by measuring the trihydroxy metabolites which were unchanged. Furthermore, the formation of glutathione derivatives of leukotriene A4 can be excluded due to the lack of glutathione transferase activity in PMNL. In order to link the intracellular function of L-ascorbate with the serum level, the ascorbate uptake has been studied in more detail. The L-ascorbate transport into PMNL is stereospecific and can best be described by kinetics consisting of a saturation part, Km and Vmax being 39 microM and 0.28 nmol/10(8) cells.min, respectively, plus passive diffusion, the diffusion coefficient P being 1.75 microliter/10(8) cells.min. Furthermore, the uptake is inhibited by the isomers D-ascorbic acid and D-erythorbic acid.(ABSTRACT TRUNCATED AT 250 WORDS)

Arachidonate 5-Lipoxygenase↗