Biomedical subjects

T Lund

Publications and source records attributed to T Lund.

At least 73 records · Page 4Linked to original sources

PubMed1998

Midazolam-fentanyl-isoflurane anaesthesia is suitable for haemodynamic and fluid balance studies in pigs.

The domestic pig is a useful model in certain areas of biomedical research. Effective use of this species is often encumbered by lack of reference values in conscious as well as anaesthetized animals. Anaesthesia itself influences physiological and biological variables; the anaesthetic technique often affects experimental results. The relationship between anaesthesia and haemodynamics is well characterized in man, but less established in pigs. We studied the effect of midazolam-fentanyl-isoflurane anaesthesia in six immature, male, domestic pigs (Norwegian landrace). Haemodynamic variables (heart rate, arterial systolic, mean, diastolic pressures, pulmonary systolic, mean, diastolic pressures, pulmonary capillary wedge pressure), tissue perfusion, lymph flow (thoracic duct) were recorded for 3 h in animals with open chest through midline sternotomy. Variables relevant to fluid balance, e.g. interstitial hydrostatic pressure (Pi), serum-colloid osmotic pressure (s-COP) and serum-albumin (s-albumin) and -protein (s-protein) concentrations were measured. With the chosen anaesthetic technique haemodynamic variables, including lymph flow, and laboratory variables remained constant during the study period. Most variables were similar to conditions in humans. In contrast to adult humans exposed to the same anaesthetic technique, these pigs had lower haemoglobin-, s-albumin- and s-protein concentrations. A finding which may reflect immaturity. Liver and lung perfusion decreased significantly during the study period whereas perfusion of the other organs studied remained constant. Lack of responses to defined noxious stimuli during the study period suggest adequate analgesia. We conclude that midazolam, fentanyl and isoflurane provide cardiovascular stability including normal microvascular fluid exchange, which are essential elements for securing the quality of results obtained during cardiovascular research in anaesthetized pigs.

Anesthesia↗

PubMed1998

Sampling of interstitial fluid and measurement of colloid osmotic pressure (COPi) in pigs: evaluation of the wick method.

The wick method for sampling of interstitial fluid from subcutis was applied in fluid balance studies in young pigs. Colloid osmotic pressure was measured in serum (COPs) and interstitial fluid (COPi) using a membrane colloid osmometer. Our aims were to determine the 'true' COPi, and to find the optimal duration of wick implantation. In series I (n = 6) a 'crossover' experiment was performed using wicks soaked in different priming solutions (non-diluted and diluted serum protein solutions or isotonic salt solution). Circulatory arrest was induced just before wick insertion in order to eliminate the vascular part of the acute inflammation. In series II (n = 6) wicks were removed in sequence after 60, 90, 120 and 180 min sampling time in anaesthetized pigs in vivo. COPs, COPi and haematocrit (HCT) together with haemoglobin (Hgb), serum albumin and total protein concentrations were determined in the same animals. In series I average COPs and COPi were 13.7 (1.4) and 7.2 (1.4) mmHg respectively (SD). In series II the optimal wick implantation times were estimated to be 60-90 min for wicks soaked in diluted protein solution, and 90-120 min for dry and saline-soaked wicks. COPs averaged 13.0 (0.7) mmHg, HCT 30.0 (1.6)%, Hgb 8.3 (0.9) g/dl, s-albumin 22.7 (0.6) g/l and s-protein 47.3 (2.3) g/l. Compared to commonly reported reference values, we found surprisingly low values for most of the measured variables. This may be related to the fact that we used immature pigs. An analysis of the validity of the wick method based on our own results and published reports is presented. We conclude that sampling of interstitial fluid with subcutaneous wicks is easy to perform in young pigs. However, the COP-values measured in wick fluid have to be carefully evaluated especially when sampling is performed in vivo.

Anesthesia↗

PubMed1998

Interbody cage stabilisation in the lumbar spine: biomechanical evaluation of cage design, posterior instrumentation and bone density.

We performed a biomechanical study on human cadaver spines to determine the effect of three different interbody cage designs, with and without posterior instrumentation, on the three-dimensional flexibility of the spine. Six lumbar functional spinal units for each cage type were subjected to multidirectional flexibility testing in four different configurations: intact, with interbody cages from a posterior approach, with additional posterior instrumentation, and with cross-bracing. The tests involved the application of flexion and extension, bilateral axial rotation and bilateral lateral bending pure moments. The relative movements between the vertebrae were recorded by an optoelectronic camera system. We found no significant difference in the stabilising potential of the three cage designs. The cages used alone significantly decreased the intervertebral movement in flexion and lateral bending, but no stabilisation was achieved in either extension or axial rotation. For all types of cage, the greatest stabilisation in flexion and extension and lateral bending was achieved by the addition of posterior transpedicular instrumentation. The addition of cross-bracing to the posterior instrumentation had a stabilising effect on axial rotation. The bone density of the adjacent vertebral bodies was a significant factor for stabilisation in flexion and extension and in lateral bending.

Absorptiometry, Photon↗

PubMed1998

Immunological analysis of epitopes on hCG.

The heterodimeric glycoprotein hormone, human chorionic gonadotropin, has been extensively characterized in terms of its recognition by mouse monoclonal antibodies. A number of different approaches have led to the definition of several epitope clusters on the surface of the molecule. These include epitopes located solely on the alpha- or beta-chain, some of which are masked when the two chains associate to form the holo-hormone. Additional epitopes comprise amino acids contributed by both the chains. In contrast to the extensive knowledge regarding B cell epitopes, the characterization of T cell epitopes on hCG has only recently begun to be explored.

Animals↗

PubMed1998

Cellular activation in response to physical exercise: the effect of platelets and granulocytes on monocyte reactivity.

After physical exercise monocytes have been shown to express more lipopolysaccharide (LPS)-induced tissue factor activity compared with monocytes at rest. This is in contrast to LPS-induced tumor necrosis factor (TNF)-alpha production, which is reduced after physical exercise. Furthermore, LPS-induced tissue factor activity in monocytes at rest has been shown to be dependent on platelets and granulocytes, whereas LPS-induced TNF-alpha is not affected by these cells. The aim of the present study was to test how platelets and granulocytes affect LPS-induced tissue factor activity in monocytes observed after exercise. This was done by separating the different blood cells before and after exercise, followed by recombination of the cells in plasma and quantification of LPS-induced tissue factor activity in vitro. Two groups of subjects with different aerobic endurance levels were tested to reveal possible differences as a result of physical conditioning. The results of the present study showed that the major effect of exercise was associated with a change in the reactivity of monocytes themselves, since monocytes from blood samples taken after running recombined with platelets, and granulocytes showed the greatest increase in tissue factor activity. Platelets and granulocytes stressed by physical exercise had no further enhancing effect on LPS-induced tissue factor activity other than that observed at rest. The lowest LPS-induced tissue factor activity in all combinations of blood cells was found in athletes, compared with less trained subjects, both at rest and in response to exercise.

Adult↗

PubMed1997

Bacillus cereus and its food poisoning toxins.

Bacillus cereus is becoming one of the more important causes of food poisoning in the industrialised world. It produces one emetic toxin and three different enterotoxins. The emetic toxin is a ring-shaped structure of three repeats of four amino and/or oxy acids: [D-O-Leu-D-Ala-L-O-Val-L-Val]3. This ring structure has a molecular mass of 1.2 kDa, and is chemically closely related to the potassium ionophore valinomycin. Two of the three enterotoxins have been shown to be involved in food poisoning. They both consist of three different proteins that act together. One of these enterotoxins is also a haemolysin. This haemolytic enterotoxin is transcribed from one operon. The third enterotoxin is a single component protein, but has not been shown to be involved in food poisoning.

Amino Acid Sequence↗

PubMed1997

Fluid resuscitation following a burn injury: implications of a mathematical model of microvascular exchange.

A validated mathematical model of microvascular exchange in thermally injured humans has been used to predict the consequences of different forms of resuscitation and potential modes of action of pharmaceuticals on the distribution and transport of fluid and macromolecules in the body. Specially, for 10 and/or 50 per cent burn surface area injuries, predictions are presented for no resuscitation, resuscitation with the Parkland formula (a high fluid and low protein formulation) and resuscitation with the Evans formula (a low fluid and high protein formulation). As expected, Parkland formula resuscitation leads to interstitial accumulation of excess fluid, while use of the Evans formula leads to interstitial accumulation of excessive amounts of proteins. The hypothetical effects of pharmaceuticals on the transport barrier properties of the microvascular barrier and on the highly negative tissue pressure generated postburn in the injured tissue were also investigated. Simulations predict a relatively greater amelioration of the acute postburn edema through modulation of the postburn tissue pressure effects.

Burns↗

PubMed1997

Can epitope-focused vaccines select advantageous immune responses?

One of the great success stories of preventive medicine is the achievement of protection against many pathogens by the simple procedure of vaccination. The rationale behind vaccination is to generate a protective immune response and an expanded population of memory cells ready to encounter the infectious agent, which will then elicit a potent secondary immune response. However, the development of effective vaccines against many pathogens has, so far, been unsuccessful. Many vaccines in current use fail to direct the immune response towards the epitopes that will ensure optimal protection. In these circumstances, can vaccines be produced that focus the immune system in a calculated, epitope-specific manner?

Animals↗

PubMed1997

Comparison of nucleic acid and protein immunization for induction of antibodies specific for HIV-1 gp120.

We have compared the antibody response to HIV-1 gp120 type LAI in mice immunized with either a gp120 expression plasmid or with baculovirus-derived recombinant gp120 (rgp120) formulated with Freund's complete adjuvant. TiterMax, Alum, Ribi R-700, AF-A or QuilA. DNA immunization resulted in variable levels of antibody, with endpoint titres ranging from 10(4) to 10(5), whereas mice immunized with rgp 120 mixed with Ribi R-700, AF-A or QuilA produced antibody levels with endpoint titres > 10(5). Both types of immunization failed to elicit antibodies able to recognize denatured rgp120. The V3 region was immunogenic in animals immunized with nucleic acid, whereas only a few animals immunized with recombinant protein produced antibodies specific for V3 or other linear epitopes, irrespective of the adjuvant used. These data suggest that the immunogenicity of gp120 is dependent upon the mode of antigen delivery, and that in vivo expressed gp120 following nucleic acid immunization elicits, at least with respect to V3, an antibody response which more closely reflects that seen following natural infection in man.

Adjuvants, Immunologic↗

PubMed1997

In vitro antiviral drug sensitivity of the Kaposi's sarcoma-associated herpesvirus.

OBJECTIVE: Kaposi's sarcoma-associated herpesvirus (KSHV), or human herpesvirus 8, has been implicated as the causative agent of Kaposi's sarcoma. Retrospective studies show that the risk of development of Kaposi's sarcoma is significantly lower in AIDS patients who received ganciclovir or phosphonoformic acid (PFA) therapy. Therefore, in vitro antiviral drug sensitivity of KSHV was studied. METHODS: The KSHV genome is a latent episome in lymphoma cells such as the BCBL-1 cell line. Lytic KSHV DNA synthesis is induced by the phorbol ester 12-O-tetradecanoyl phorbol-13-acetate in BCBL-1 cells; this system was used to evaluate the effects of antiviral drugs on KSHV DNA synthesis. RESULTS: Linear (lytic) KSHV DNA synthesis and virus secretion was inhibited in BCBL-1 cell cultures by cidofovir (median inhibitory concentration, 0.05 microM), ganciclovir (5.1 microM) and PFA (97 microM), and by aciclovir (75 microM). Prolonged incubation of BCBL-1 cells with antiviral drugs had no effect on episomal KSHV DNA synthesis. CONCLUSIONS: The antiviral drug assay developed shows that KSHV is very sensitive to cidofovir, moderately sensitive to ganciclovir and PFA, and weakly sensitive to aciclovir. Therefore, low doses of cidofovir, or high doses of PFA or ganciclovir could suppress clinical reactivation of KSHV. Antiviral drugs did not inhibit episomal virus DNA synthesis, suggesting that the latent form of viral DNA is replicated by host DNA polymerases. Consequently, no benefit can be expected from antiviral drugs in KSHV-positive B-cell lymphomas or during latency.

Acyclovir↗

PubMed1997

Herpesvirus saimiri Tip-484 membrane protein markedly increases p56lck activity in T cells.

Herpesvirus saimiri (HVS) is a T-cell-specific transforming and oncogenic virus. A protein encoded by HVS known as Tip-484 (for tyrosine kinase interacting protein from HVS strain 484) is required for this transformation. Tip-484 binds specifically to the nonreceptor protein tyrosine kinase p56lck. By transfecting Tip-484 into T cells, we now show that this interaction leads to a several hundred-fold increase in the kinase activity of p56lck. Tip-484 is part of a protein complex which is dependent on the presence of p56lck and is phosphorylated. We also show that two of the complexed proteins represent two phosphorylated forms of Tip-484. Furthermore, the p56lck kinase activity in HVS-infected human peripheral blood T lymphocytes was at least ninefold higher than that in noninfected control cells and significantly decreased in cells infected with a Tip-484 deletion mutant virus. Finally, we report that Tip-484 is required for oncogenesis in rabbits by the survival of rabbits inoculated with Tip-484 deletion mutant HVS. The data demonstrate dramatic stimulation of the signaling pathway of p56lck. This effect can contribute to the molecular mechanisms that lead to sustained autocrine secretion of growth factors, permanent T-cell growth, and ultimately lymphocytic tumor formation.

Animals↗

PubMed1996

Immunochemical mapping of gonadotropins.

As a glycoprotein hormone, human chorionic gonadotropic (hCG) is not a single molecular entity but this term rather comprises an array of molecular variants such as hCG, hCG beta, hCGn, hCG beta n, hCG beta cf, -CTPhCG, hCG beta CTP, deglyhCG, asialohCG, hCGav and the closely related molecules hLH, hLH beta and hLH beta ef. The advent of monoclonal antibodies (MCA), the availability of ultrasensitive detection systems and the recent determination of the crystal structure of hCG, made it possible to design special purpose diagnostic and clinical research immunoassays for hCG-like molecules. For more than a decade we and others have tried to refine epitope maps for hCG and related molecules by means of a large panel of MCA, naturally occurring metabolic variants of hCG (hCGn, hCG beta, hCG alpha, hCG beta cf, hCG beta CTP), homologous hormones and subunits of various species (e.g. hLH, hLH beta, hFSH, hTSH, oLH, rLH beta), chemically modified molecules (deglyhCG, asialohCG, tryptic and chymotryptic hCG beta and hCG alpha fragments) and synthetic peptides (octapeptides and longer). It appeared that all epitopes on molecular hCG-variants recognized by our MCA are determined by the protein backbone. Except for the two major epitopes on hCG beta CTP and parts of two antigenic domains on hCG alpha, epitopes on hCG-derived molecules are determined by the tertiary and quarternary structure. Operationally useful descriptive epitope maps were designed including information on assay suitability of antigenic determinants. On this basis we established ultrasensitive time-resolved fluoroimmuno-assays for hCG, hCG and hCGn, hCG beta and hCG beta n and hCG beta cf, hCG alpha and additional assays recognizing different spectra of hCG-variants. Such assay have been applied by us and others to the detection of pregnancy, early pregnancy loss, choriocarcinoma, testicular cancer, other cancers and prenatal diagnosis. However, as the molecular structure of many epitopes utilized in immunoassays of different laboratories was not resolved, comparability of results was not satisfactory. Consequently, attempts were made to compare schematic epitope maps from different research institutions. The situation has been much improved by solving the three-dimensional (3D) structure of hCG. It has been shown that hCG is a member of the structural superfamily of cystine knot growth factors like NGF, PDGF-B and TGF-beta. Each of its subunits is stabilized in its topology by three disulfide bonds forming a cystine knot. Moreover, it turned out that the disulfide bridges in their majority have previously been wrongly assigned. Computer molecular modeling of crystallographic coordinates of hCG and subsequent selective combined--PCR-based and immunological--mutational analyses of hCG beta expressed via the transmembrane region of a MHC molecule made it possible to more precisely localize epitopes on hCG-derived molecules. Although the entire surface of hCG has to be regarded as potentially immunogenic there seems to be hot spots where epitopes are clustered in antigenic domains. These are located on the first and third loops protuding from the cystine knots of both subunits and are possibly centered around the knot itself. Ultimate answers on epitope localizations will be given by the crystal structure determination of hCG complexed with different Fabs.

Amino Acid Sequence↗

PubMed1996

The relative importance of vertebral bone density and disc degeneration in spinal flexibility and interbody implant performance. An in vitro study.

STUDY DESIGN: An in vitro biomechanical investigation in the human lumbar spine focuses on the functional significance of vertebral bone density and intervertebral disc degenerations. OBJECTIVE: To determine that interrelationship between vertebral bone density and intervertebral disc degeneration, their effect on normal spine motion, and their significance in the biotechnical performance of interbody fixation techniques. SUMMARY OF BACKGROUND DATA: A relationship between vertebral bone density and intervertebral disc degeneration has been suggested, but a definitive relationship has not been established. The effect of vertebral bone density and intervertebral disc degeneration on interbody stabilization remains unknown despite the rapidly increasing use of this surgical method for patients with chronic low back pain. METHODS: The vertebral bone density and intervertebral disc degeneration of 72 functional spinal units were determined using dual energy x-ray absorptiometry scans and macroscopic grading, respectively. A three-dimensional flexibility test was performed on 24 functional spinal units in the intact and stabilised conditions. The compressive behavior of the bone-implant interface was evaluated in 48 functional spinal units. RESULTS: The vertebral bone density in moderately degenerated disc was significantly lower than at all other levels of intervertebral disc degeneration. Increasing intervertebral disc degeneration resulted in more axial rotation and less lateral bending. In flexion-extension and lateral bending, better vertebral bone resulted in significantly better stabilization. This trend was observed also in axial compression in which higher failure loads were observed with greater bone densities. CONCLUSION: The authors conclude a significant relationship exists between bone density and disc degeneration, bone density is a highly important factor in the performance of interbody stabilization, and disc degeneration, is of moderate importance in signal motion.

Biomechanical Phenomena↗

PubMed1996

Evidence for a further enterotoxin complex produced by Bacillus cereus.

Out of 321 strains of Bacillus cereus from several sources and isolated in four different countries, 239 (74%) produced cytotoxins. Only 127 (53%) of the cytotoxic strains were positive for the B-component gene of the haemolysin BL (enterotoxin) by polymerase chain reaction (PCR). Western blots using antiserum produced against enterotoxin(s) gave positive results for 199 (83%) of the cytotoxic B. cereus strains. On closer examination of seven of the strains, involved in food poisoning, we found that two strains completely lacked the L2- and B-components (of the haemolysin BL), and two strains were negative for the B-component gene by PCR, but were positive for the L2-component. From our experiments we concluded that there is at least one enterotoxin complex in addition to the haemolysin BL enterotoxin and enterotoxin T.

Animals↗

PubMed1996

Characterisation of a non-haemolytic enterotoxin complex from Bacillus cereus isolated after a foodborne outbreak.

Three enterotoxic components have been isolated from a strain of Bacillus cereus which was involved in a large food poisoning outbreak in Norway in 1995. The components were purified by chromatography on three different columns. Three proteins of 39, 45 and 105 kDa, respectively, were found to be necessary for maximum cytotoxicity. The amino acid N-terminal sequences of the 39 and 45 kDa proteins were determined. The 45 kDa component was the same protein as the main antigen detected in the Bacillus Diarrhoeal Enterotoxin Visual Immunoassay (Tecra). The 39 kDa protein showed some similarity to the L1 protein of haemolysin BL from B. cereus. Furthermore, the three toxic components were all recognised by a polyclonal antiserum reported to detect enterotoxin from B. cereus. The proteins were different from the B- and L2-components of haemolysin BL, previously suggested to be a primary virulence factor, and had no detectable haemolytic activity.

Amino Acid Sequence↗

PubMed1996

Relative location of epitopes involved in synergistic antibody binding using human chorionic gonadotropin as a model.

We systematically screened a large panel of well-characterized monoclonal antibodies (mAb) directed towards various epitopes on human chorionic gonadotropin (hCG) for synergistic binding of 125I-hCG when they were adsorbed to a solid phase. The epitope locations involved in synergy were then related to the crystal structure of hCG and discussed in accordance with available data on the hCG epitopes. Enhanced binding of hCG was specific for certain pairs of mAb and was reflected in a 3-50-fold increased apparent functional affinity constant for hCG. Surface plasmon resonance revealed that when the mAb were captured by a polyclonal anti-IgG1 coupled to the Biacore chip, the off rates for hCG were significantly slower with synergistic mAb combinations than for the corresponding single mAb or nonsynergistic pairs of mAb, whereas the on rates did not differ appreciably. Each of the two antibodies involved in synergistic binding of hCG (more than 3-fold compared to additive binding of the two mAb) always belonged to a different epitope cluster in a separate antigenic domain on hCG. Synergistic epitope combinations on holo-hCG were located in similar structural planes. Combinations of mAb directed towards the epitope clusters alpha 2/beta 3/5, alpha 2/hCG beta CTP (C-terminal peptide) and beta 3/5/hCG beta CTP showed the strongest enhancement, with binding more than 10-fold greater than the sum of 125I-hCG bound to the individual mAb, followed by pairs of mAb directed towards the epitope groups beta 1/beta 3/5, c 1/2/beta 3/5, beta 1/alpha 2, and alpha 2/alpha 3/5 (3-9-fold). The greater frequency of synergy obtained with the linear epitopes of the hCG beta CTP can be ascribed to their greater molecular flexibility relative to the constrained discontinuous epitopes on hCG alpha and core-hCG beta (residues 1-112). In general, these studies provide a method for rapid screening of synergistic antibody pairs which also helps to identify non-overlapping epitopes that are accessible in similar structural planes. In turn, this facilitates the design of high-affinity bispecific antibodies targetted to a single antigen molecule.

Antibodies, Monoclonal↗

PubMed1996

Identification and selective destruction of shared epitopes in human chorionic gonadotropin beta subunit.

The feasibility of producing epitope-specific antigens by mutation of the gene is demonstrated, the aim being to eliminate unwanted surface epitopes yet allowing the natural folding of the protein to maintain the desired epitope(s). The model protein is the beta subunit of human chorionic gonadotropin (hCG beta) which previously has been used as an immunological contraceptive vaccine but has extensive cross-reaction with human luteinizing hormone. Of a series of mutants made, the mutant with substitutions of Glu for Arg 68, Ser for Arg 74, His for Gly 75 and His for Val 79, lost the ability to react with a panel of cross-reacting monoclonal antibodies while retaining the discontinuous and linear epitopes specific to the holo-hormone. In addition, allocation of amino acid residues to established epitope clusters could be made: residues 24, 25, 68 and 71 probably contribute to the cluster termed beta 3, residues 20, 21, 22, 75 and 77 to cluster beta 6 and residue 68 to clusters beta 2, beta 4 and beta 5.

Chorionic Gonadotropin, beta Subunit, Human↗

PubMed1996

Comparison of techniques for reinforcing stone molds used to construct facial prostheses.

Dental stone molds are convenient for fabricating facial prostheses, but they tend to break when repeatedly used to make duplicate prostheses. This study investigated the use of an inexpensive plastic mesh, die hardener, and PVC pipe segment to reinforce stone mold test samples that were broken in compression on an Instron testing machine. The samples reinforced with PVC pipe surpassed the load limit of the Instron machine. The samples reinforced with both die hardener and mesh were 13% more resistant to breakage than the control samples made of stone alone. Samples reinforced only with mesh were 3% weaker than the control samples.

Analysis of Variance↗