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Biomedical subjects

T Thorsen

Publications and source records attributed to T Thorsen.

At least 19 recordsLinked to original sources

Monolithic microfabricated valves and pumps by multilayer soft lithography.

Soft lithography is an alternative to silicon-based micromachining that uses replica molding of nontraditional elastomeric materials to fabricate stamps and microfluidic channels. We describe here an extension to the soft lithography paradigm, multilayer soft lithography, with which devices consisting of multiple layers may be fabricated from soft materials. We used this technique to build active microfluidic systems containing on-off valves, switching valves, and pumps entirely out of elastomer. The softness of these materials allows the device areas to be reduced by more than two orders of magnitude compared with silicon-based devices. The other advantages of soft lithography, such as rapid prototyping, ease of fabrication, and biocompatibility, are retained.

Adhesiveness↗

[Student education in a laboratory for general practice skills].

Clinical skills, like questioning and examining a patient, are developed gradually throughout the years of medical training. Basic skills should be mastered on graduation, but the teaching of skills in most medical schools is not systematic. We evaluated a pilot teaching project in a general practice (GP) skills laboratory. Students were randomised to an intervention group and a control group. Teaching was performed in small groups of one GP instructor and six students and consisted of four weekly three-hour sessions. For the control group, teaching was delayed until after evaluation. Evaluation of practical skills and communication skills was made with the students in random order by GP evaluators who did not know about the randomisation procedure. Each skill was scored as satisfactory or not satisfactory for each of five procedural elements. 56 fourth-year medical students volunteered. Evaluation was performed for 19 (68%) students in the training group and 18 (64%) in the control group. The training group performed better on practical skills, average score 9.7 (95% confidence interval 7.4 to 12.1) vs. 5.5 (3.2 to 7.8), (p = 0.01). There was no significant difference for communication skills, 7.7 (5.6 to 9.8) vs. 6.7 (4.6 to 8.7). Teaching clinical skills in a general practice skills laboratory is feasible. Practical skills may improve rapidly, at least for short-time performance, while our brief teaching in communication skills was not proved to be effective.

Adult↗

Changing doctor prescribing behaviour.

The aim of this overview was to identify interventions that change doctor prescribing behaviour and to derive conclusions for practice and further research. Relevant studies (indicating prescribing as a behaviour change) were located from a database of studies maintained by the Cochrane Collaboration on Effective Professional Practice. This register is kept up to date by searching the following databases for reports of relevant research: DHSS-DATA; EMBASE; MEDLINE; SIGLE; Resource Database in Continuing Medical Education (1975-1994), along with bibliographies of related topics, hand searching of key journals and personal contact with content area experts. Randomised controlled trials and non-equivalent group designs with pre- and post-intervention measures were included. Outcome measures were those used by the study authors. For each study we determined whether these were positive, negative or inconclusive. Positive studies (+) were those that demonstrated a statistically significant change in the majority of outcomes measured at level of p < or = 0.05 between the intervention and control groups. Negative studies (-) showed a significant change in the opposite direction and inconclusive studies (approximately) showed no significant change compared to control or no overall positive findings. We identified 79 eligible studies which described 96 separate interventions to change prescribing behaviour. Of these interventions, 49 (51%, 41%-61%) showed a positive significant change compared to the control group but interpretation of specific interventions is limited due to wide and overlapping confidence intervals.

Data Collection↗

High-throughput genetic analysis using microfabricated 96-sample capillary array electrophoresis microplates.

Capillary array electrophoresis (CAE) microplates that can analyze 96 samples in less than 8 min have been produced by bonding 10-cm-diameter micromachined glass wafers to form a glass sandwich structure. The microplate has 96 sample wells and 48 separation channels with an injection unit that permits the serial analysis of two different samples on each capillary. An elastomer sheet with an 8 by 12 array of holes is placed on top of the glass sandwich structure to define the sample wells. Samples are addressed with an electrode array that makes up the third layer of the assembly. Detection of all lanes with high temporal resolution was achieved by using a laser-excited confocal fluorescence scanner. To demonstrate the functionality of these microplates, electrophoretic separation and fluorescence detection of a restriction fragment marker for the diagnosis of hereditary hemochromatosis were performed. CAE microplates will facilitate all types of high-throughput genetic analysis because their high assay speed provides a throughput that is 50 to 100 times greater than that of conventional slab gels.

Capillary Action↗

Puncture technique and postural postdural puncture headache. A randomised, double-blind study comparing transverse and parallel puncture.

BACKGROUND: This clinical study was conducted in order to investigate the effect of two different orientations of the bevel during dural puncture on development of postural postdural puncture headache (PPDPH). METHODS: Two hundred and eighteen patients aged 18 to 50 years scheduled for minor non-obstetric surgery using spinal anaesthesia (SA) were included in this randomised, double-blind study. Dural puncture was performed using a 0.42 mm O.D. (27-g) Quincke spinal needle with the orientation of the bevel parallel or transverse relative to the longitudinal axis of the dural cylinder. All patients were blinded with regard to the puncture technique, and so was the anaesthesiologist performing a telephone interview 5 to 7 days postoperatively. The occurrence and duration of headache, backache and other complaints were recorded. Headache was classified as PPDPH or non-PPDPH, and intensity of the headache was registered using a numerical rating scale (NRS) from 0 to 10. RESULTS: Two hundred and twelve patients with a mean age of 35.3 years completed the study, 106 in each group. The two groups were comparable with regard to mean age, sex, local anaesthetics used and surgical procedure performed. Headache occurred in 44 patients postoperatively. PPDPH was diagnosed in 4/106 patients (3.8%) in the parallel group and 24/106 (22.6%) in the transverse group (P < 0.0002). Postoperative backache occurred in 31 and 20 patients (parallel compared to transverse) (NS). CONCLUSIONS: Dural puncture with the bevel of the needle transverse to the longitudinal axis of the dural cylinder gave significantly more cases of PPDPH than puncture with the bevel parallel to this axis even when using a 27-g Quincke needle. When using Quincke bevelled needles care must be taken to assure that the orientation of the bevel is parallel to the longitudinal axis of the dural sac.

Adolescent↗

Modulation of mouse estrogen receptor transcription activity by protein kinase C delta.

The effect of protein kinase C (PKC) delta on the transcriptional activity of the mouse estrogen receptor was investigated. The receptor was expressed transiently in Cos-1 and NIH3T3 cells in the presence of wild-type, dominant negative or constitutively active forms of PKC delta. Transfection experiments demonstrated that PKC delta stimulated both unliganded and liganded estrogen receptor transcriptional activity. This stimulatory effect was not observed using PKC alpha or PKC epsilon. 4-Hydroxytamoxifen and the pure anti-estrogen ICI 164,384 reduced receptor transcriptional activity in the presence of PKC delta. The stimulatory effect of PKC delta on estrogen receptor transcriptional activity was mediated by the N-terminal activation function 1 (AF-1) domain. The reduced stimulatory effect of PKC delta on transcriptional activity of the phosphorylation defective mutant of estrogen receptor suggests that phosphorylation of serine 122 in the AF-1 region may mediate the modulatory effect of PKC delta. Wild-type PKC delta caused a twofold increase in estrogen receptor phosphorylation, while a dominant negative mutant of PKC delta reduced the receptor phosphorylation to five percent of that caused by wild-type PKC delta. Our results suggest that PKC delta participates in the signaling pathways that lead to estrogen receptor phosphorylation and its effect on estrogen receptor transcriptional activation is both cell type and promoter specific.

3T3 Cells↗

Eicosapentaenoic acid and sulphur substituted fatty acid analogues inhibit the proliferation of human breast cancer cells in culture.

Numerous studies have shown dietary fatty acids to influence the progression of several types of cancers. The purpose of the present investigation was to examine the influence of various types of fatty acids, including omega-3 fatty acids and a new class of hypolipidemic peroxisome proliferating fatty acid analogues, namely the 3-thia fatty acids, on MCF-7 human breast cancer cell growth. 3-thia fatty acids represent non-beta-oxidizable fatty acid analogues in which a sulphur atom substitutes for the beta-methylene group (3-position) in the saturated and unsaturated fatty acids. The effects of increasing concentrations of palmitic acid, tetradecylthioacetic acid (a 3-thia fatty acid), eicosapentaenoic acid, docosahexaenoic acid, and two 3-thia polyunsaturated fatty acids on the proliferation of MCF-7 cells, maintained in serum-free culture, were studied. At the highest concentration of fatty acid used (64 microM) tetradecylthioacetic acid was found to be the most effective of all fatty acids tested in inhibiting cell growth, whilst palmitic acid and docosahexaenoic acid had no significant effect on cell growth. Thus, of the two dietary polyunsaturated omega-3 fatty acids eicosapentaenoic acid and docosahexaenoic acid, only eicosapentaenoic acid possesses an inhibitory effect on the proliferation of MCF-7 cells. In all cases the inhibitory effect of the fatty acid was found to be reversible. Tetradecylthioacetic acid has been shown to be a potent peroxisome proliferator. It was, therefore, hypothesized that tetradecylthioacetic acid may inhibit the human MCF-7 cell growth by increasing the level of oxidative stress within the cell. However, use of agents which modify the cell's protective apparatus against oxidative stress had no influence on the inhibitory effect of tetradecylthioacetic acid. These experiments indicate that tetradecylthioacetic acid inhibits cell growth by mechanisms which may be independent of oxidative status.

Antimetabolites, Antineoplastic↗

Follow-up after torsion of the spermatic cord.

Fifty-two patients were examined 4-10 1/2 years after operation for torsion of the spermatic cord. The testicular salvage rate was 56%. Loss of testicular tissue was correlated to preoperative duration of symptoms. Serum testosterone concentration was found to be within the reference range. FSH was elevated in patients with the longest duration of symptoms (> 24 h), suggesting an impairment of testicular function in these patients. There were two our of 13 patients with a severe oligo-azoospermia, both had a previous orchiopexy for undescended contralateral testis.

Adolescent↗

Evaluation of thiopentone-midazolam-fentanyl anaesthesia in pigs.

Induction of anaesthesia in swine by thiopentone (27.1-35.7 mg/kg, mean 29.9 mg/kg) was followed by bolus doses and continuous infusion of midazolam and fentanyl (0.90 mg/kg followed by 0.90 mg/kg/h and 0.025 mg/kg followed by 0.025 mg/kg/h, respectively). This produced good anaesthesia and analgesia for up to 2 h in 6 Norwegian Landrace pigs (wt: 17-42 kg), based on responses to painful stimuli elicited by pinching the nasal septum, the mouth, the forefoot and the perineal skin area. The first responses occurred after 110 min of anaesthesia. No significant drop in rectal temperature due to the regimen was noted during monitoring periods (140-180 min). This combined intravenous anaesthetic regimen gave good anaesthesia and analgesia to pigs for up to 2 h as monitored by clinical signs. The regimen may not be sufficient for longer time periods. We cannot advocate the incorporation of neuromuscular blocking agents in this regimen.

Anesthesia, Intravenous↗

A transport and monitoring unit for piglets under general anaesthesia.

A transport and monitoring unit for fully anaesthetized piglets (weight 20-30 kg) was made. The unit provided easy transport for short or longer distances while at the same time making continuous monitoring of invasive blood pressure (IBP), temperature and HR possible. The animals stayed in the unit for the duration of the experiments which required that the anaesthetized animal be kept in exactly the same position for several hours. This, as well as the experimental animal's welfare, was ensured by the unit. Such a unit can be used for any lengthy transportation of experimental animals to research facilities located separately from animal laboratories.

Anesthesia, General↗

Nitrogen microbubbles induce a disappearance of single platelets (aggregation) with porcine platelets: a comparative study of the effects of anticoagulants and blood collection methods.

The pathogenesis of decompression illness (DCI) is uncertain. DCI involves all parts of the organism where gas bubbles are produced. They have both primary and secondary effects and have been classified as an agonist aggregating human platelets. In vitro effects of N2 bubbles on porcine platelets were investigated. Comparative studies using two different anticoagulants and three different sampling methods were performed. A disappearance of single platelets interpreted as platelet aggregation was observed in the presence of N2 bubbles in all studied groups. Aggregatory responses were more profound with platelets in heparinized plasma than in citrated plasma. In citrated plasma the aggregatory responses were more profound when blood was obtained from nonanaesthetized (awake) animals than from slaugtherhouse animals. Adrenaline (1 microM) had an inhibitory effect on N2 bubble induced platelet aggregation in vitro. The pig could be useful to investigate possible gas bubble effects in vivo.

Animals↗

Microbubble-induced phospholipase C activation does not correlate with platelet aggregation.

The effect of nitrogen-(N2-)microbubbles on platelets resembles that of common platelet agonists with respect to aggregation and secretion, but is considerably slower and is poorly inhibited by aspirin. This paper reports the effect of microbubbles on platelet phospholipase C activity in gelfiltered human platelets prelabelled with [32P]Pi ([32P]-GFP). The experiments were run in the presence of an ADP scavenging system in order to rule out effects of ADP. Stimulation of [32P]-GFP for 30 min with microbubbles caused a significant reduction in single platelets (p < 0.0004) and a significant increase in 32P-activity in the phosphatidic acid (PA) fraction (p < 0.02). Epinephrine potentiated the microbubble-induced reduction in single platelets (p < 0.05), but did not enhance the amount of 32P in the platelet [32P]PA fraction. The 32P-radioactivity in the PI-fraction increased with time to a similar extent when [32P]-GFP was stirred for 30 min in absence of microbubbles as it did after 30 min of agonist exposure. There were no significant changes in the [32P]PIP and [32P]PIP2 fractions. Aspirin abolished the microbubble-induced increase in 32P-activity in the PA fraction, but had no significant effect on the reduction in single platelets. Aspirin had a small but significant, reducing effect on platelet aggregation induced by a combination of epinephrine and microbubbles (p < 0.05). With epinephrine, however, aspirin did not completely abolish the increase in [32P]-PA. It is concluded that microbubbles alone cause platelets to aggregate by a novel mechanism that operates independent of cyclooxygenase-dependent arachidonic acid metabolites and phospholipase C activation.

Aspirin↗

Bubble-induced aggregation of platelets: effects of gas species, proteins, and decompression.

We show that bubbles containing different gases (N2, He, Ne, Ar, or an O2-CO2-N2 mixture) are equally potent platelet agonists. The synergistic effect of different platelet antagonists does not seem to be affected by the type of gas in the bubbles. In contrast to aggregation in platelet-rich plasma (PRP), bubbles cause only a weak response in gel-filtered platelets (GFP), i.e., comparison of aggregation in protein-rich and protein-poor platelet suspensions may shed light on the role of different plasma proteins. Extracellular fibrinogen promotes bubble-induced platelet aggregation similar to known physiologic agonists, whereas albumin counteracts this aggregation. Bubble-induced aggregation is inhibited in GFP-fibrinogen by 2-deoxy-D-glucose plus antimycin A, suggesting dependency on ATP generation in the platelets and evidence for direct exposure of the "cryptic" fibrinogen receptor by bubbles. Hyperbaric compression and subsequent rapid, inadequate decompression of PRP caused little change in the aggregation response to gas bubbles and epinephrine at 1 bar, but reduced the response to ADP. Bubbles tended not to form before the surface film was broken. Pressure-induced aggregation was apparently metabolically active and not due to passive agglutination; electron microscopic studies and PRP with added glutaraldehyde did not show platelet activation, clumping, or reduced platelet count. In contrast to aggregation caused by pressure, bubble-induced aggregation in PRP at 1 bar (after treatment in the pressure chamber) was nearly completely inhibited by theophylline, a phosphodiesterase inhibitor that increases intracellular platelet cyclic AMP.

Adenosine Diphosphate↗

Oestradiol treatment increases the sensitivity of MCF-7 cells for the growth stimulatory effect of IGF-I.

MCF-7 cells were grown in serum free medium (Dulbecco MEM without phenol red, supplemented with Costar SF-1 without insulin). Insulin was added as required and gave dose dependent growth stimulation at concentrations between 5 and 10,000 nM. Identical growth response curves were obtained for thymidine uptake and cell number. Oestradiol and insulin-like growth factor I (IGF-I) added individually both gave a dose dependent stimulation of cell growth in serum free medium containing 50 nM insulin. The growth stimulatory effect of oestradiol was to a large extent inhibited with suramine, a general inhibitor of growth factors, indicating that the effect of oestradiol was mediated through stimulating autocrine secretion of a growth factor. To investigate a possible link between the effects of oestradiol and IGF-I, a specific IGF-I receptor antibody (alpha IR-3), 10 micrograms/ml was used. These experiments were carried out with 2.5 nM insulin in the medium, a concentration at which insulin had no growth stimulatory effect. Stimulation was carried out for 18 h before assay of thymidine uptake. The effect of oestradiol was not significantly reduced by alpha IR-3, indicating that IGF-I was not an autocrine mediator of oestradiol stimulation of cell growth under these conditions, whereas alpha IR-3 extensively reduced growth stimulation by IGF-I. On long term stimulation (5 days) oestradiol had a marked stimulatory effect on cell number and alpha IR-3 almost totally abrogated this effect. When oestradiol (1 nM) and IGF-I (2.5 nM) were added together, the combined effect on thymidine incorporation and cell number was significantly greater than additive. This synergistic effect on the IGF-I growth response was totally abolished by the IGF-I receptor antibody. The results suggest a cooperative interaction of oestradiol and IGF-I. It is concluded that growth stimulation of MCF-7 cells by long term treatment with oestradiol may be mediated through autocrine secretion of IGF-I. the effect of short term stimulation of thymidine incorporation suggest that the growth response of oestradiol is more complex, and indicate that a cooperative interaction with IGF-I is involved, which is unrelated to stimulated autocrine secretion.

Antibodies↗

Porcine platelets in vitro and in vivo studies: relevance to human thrombosis research.

This review summarizes present knowledge on porcine platelets in vitro and recent studies on in vivo activation of platelets in the pig. There are certain differences compared to human platelets: Platelet aggregation and secretion cannot be achieved by epinephrine, and the arachidonate pathway seems poorly developed in porcine platelets. Genetic models for von Willebrand disease (vWD) and storage pool deficiency (SPD) have been developed in the pig. Several models for the study of in vivo platelet deposition and early thrombus formation have been developed. Platelet radio-labeling techniques (with 111In) have been used extensively. We conclude that the pig seems to be a good choice for the investigation of in vivo platelet activation and deposition based on present knowledge of porcine platelets and on already established animal models.

Angioplasty, Balloon↗

Effects of hyperoxia on human endothelial cell proliferation and morphology in vitro.

The effects of hyperoxia on endothelial cells from human umbilical veins were studied in vitro by flow cytometric DNA analysis, cell proliferation, and morphology. Exposure of the cells to PO2 of approximately 400 and 500 mmHg (corresponding to O2 concentrations of 65 and 95%, respectively) caused, after 24 h, an increase in the proportion of S-phase cells from 18 to 33%. This was followed by a dramatic decrease of S-phase cells to about 4% after 48 h of O2 exposure. This decrease was also sustained after 72 h of O2 treatment. Concomitant with the depletion of S-phase cells was an increase from 13 to 21% in the proportion of cells in the G2M phase, indicating that cells were unable to undergo cell division after a 48-h exposure to 65 and 95% O2. Cells exposed to various PO2 levels (124, 248, 400, and 500 mmHg) showed a dose-dependent reduction in cell proliferation leading to a total inhibition of cell growth after exposure to PO2 of 400 and 500 mmHg. Cells exposed to 248 mmHg PO2 exhibited similar growth rates as the control cells during the first 3 days. Thereafter there was a 50% reduction in cell density as compared to the control cells. Light and scanning electron microscopy showed that the hyperoxic cells underwent morphologic changes characterized by cell contraction and swelling. Thus it is shown that exposure to very high PO2 tensions first stimulates the cells to enter the S phase of the cell cycle. Thereafter changes take place leading to a nearly complete reduction of cell proliferation.

Cell Count↗