Biomedical subjects

T Espevik

Publications and source records attributed to T Espevik.

153 records · Page 9Linked to original sources

PubMed1982

Porphyrin-sensitized photoinactivation of human cells in vitro.

NHIK 3025 cells derived from a carcinoma in situ were exposed to hematoporphyrin derivative (Hpd) and light and examined by light microscopy, freeze-etching, scanning, and transmission electron microscopy. The first morphologic changes observed were shrinkage of mitochondria and formation of vesicles on the cell membrane. Furthermore, increased membrane permeability led to accumulation of Hpd and cellular swelling, with a concomitant reduction in the number and size of the microvilli. Some of the vesicles seemed to originate from microvilli. The freeze-etching appearance of the membranes of the majority of the cells was unaltered by treatment with Hpd and light. However, in some cases clustering of membrane particles was observed. At low doses membrane vesiculation and cell swelling were reversed within a few hours after treatment, indicating that repair processes were operative.

Carcinoma in Situ↗

PubMed1981

A freeze-etch electron microscopic study of liquid propane jet-frozen human erythrocyte membranes.

Freeze-etch electron micrographs of haemolysing erythrocytes and isolated erythrocyte membranes frozen using a liquid propane jet-freezer reveal fracture faces very different from those seen after conventional freezing by dipping the specimens into partly solidified Freon 22. Instead of the rather smooth extracellular fracture faces found after conventional specimen freezing, extracellular fracture faces exhibiting large amounts of fibre-like structures are seen after liquid propane jet-freezing of these specimens. No such structures were found in normal red blood cells. When isolated erythrocyte membranes are frozen under conditions favouring spectrin-actin release, freeze-etch micrographs reveal an apparent continuity between the fibre-like structures on the extracellular fracture face and the long fibre-like structures which extent from the protoplasmic surface of the erythrocyte membrane. These results suggest that liquid propane jet-freezing is capable of revealing a structural difference between the membrane of haemolysing and nonhaemolysed red blood cells, and that this difference is related to the fibrous, peripheral proteins of the membrane.

Erythrocyte Membrane↗

PubMed1981

In situ liquid propane jet-freezing and freeze-etching of monolayer cell cultures.

A procedure for in situ liquid propane jet-freezing of unfixed and unglycerinated monolayer cell cultures is described. We have used the method to study monolayer cultures of rat Sertoli cells and human monocytes. The described procedure gives a cooling rate during specimen freezing high enough to yield adequately frozen specimens even in the absence of any cryoprotectant. Use of the procedure therefore eliminates artefacts usually caused by cell fixation and addition of cryoprotectives.

Animals↗

PubMed1978

A freeze-etch study of dense fibres in rat spermatozoa.

The presence of oblique striations on the cortex of the dense fibres of the rat spermatozoon is confirmed; they have a main periodicity of approximately 40 nm and a secondary one of about 20 nm. Disruption of spermatozoa by Triton X-100 and dithiothreitol and fixation in glutaraldehyde do not affect the pattern. The dense fibre cortex appears to consist of globular substructure units with a diameter of about 19 nm.

Animals↗

PubMedDate not supplied

Activation of tumor necrosis factor--alpha system in HIV-1 infection: association with markers of immune activation.

The relationships between serum levels of soluble tumor necrosis factor receptors (sTNFRs) and other prognostic and immunological parameters in different immunological subgroups of 64 HIV-1 infected patients were studied. In the patient group as a whole, the raised serum levels of sTNFRs were significantly inversely correlated to the numbers of CD4+ and CD8+ lymphocytes and significantly positively correlated with serum levels of neopterin, HIV-1 p24 antigen and the soluble CD8/CD8+ lymphocyte ratio. However, when the patients were classified into three separate immunological subgroups according to the numbers of CD4+ lymphocytes, only serum levels of neopterin were significantly correlated to levels of sTNFRs in all the defined immunological subgroups. These results indicate that HIV-1 infection is associated with a persistent and chronic immune activation in the TNF system manifested by raised serum levels of sTNFRs, which may reflect sustained activation of the immune system particularly in monocytes/macrophages. Further, these results confirm that, when comparing immunological and virological parameters in HIV-1 infection, different results may be obtained in different immunological subgroups of patients.

Adolescent↗

PubMedDate not supplied

The role of interleukin-2 in regulating the sensitivity of natural killer cells for Fas-mediated apoptosis.

The Fas/Fas-ligand (FasL) system seems to play a key role in regulating immunoresponses. Highly purified CD56+CD3- natural killer (NK) cells were found to be resistant to the apoptosis-inducing Fas mAb CH11 in the absence or in the presence of interleukin-2 (IL-2) for up to 3 days. However, NK cells activated with IL-2 for 3 days became apoptotic following combined treatment with CH11 and actinomycin D, suggesting the presence of an intact apoptotic machinery. In contrast, NK cells cultivated in IL-2 for 6 days became sensitive to CH11-induced apoptosis without addition of actinomycin D. At this time, a pronounced up-regulation of the Fas protein on the NK cell membrane was detected. By using reverse transcription/polymerase chain reaction it was found that the anti-apoptotic gene FLIP was strongly expressed in NK cells for up to 6 days of IL-2 stimulation. After day 6, a time-dependent decrease in the expression of FLIP was observed concomitantly with increased sensitivity for Fas-mediated apoptosis. The amount of apoptotic and necrotic NK cells in the presence of IL-2 increased in a time-dependent manner, reaching 40% at day 6 of culture. The amount of apoptotic and necrotic NK cells was reduced in the presence of Fas-Fc protein. In addition, IL-2 stimulated the NK cells to release soluble FasL in a time-dependent manner, whereas membrane FasL did not seem to increase in a similar manner. These results indicate that Fas/FasL interactions are involved in the down-regulation of IL-2-activated human NK cells.

3T3 Cells↗

PubMedDate not supplied

Alginate-polylysine-alginate microcapsules: effect of size reduction on capsule properties.

Alginate-polylysine-alginate capsules containing insulin-producing cells have been used as a bio-artificial pancreas in the treatment of diabetes mellitus. In a search for microcapsules with improved diffusion characteristics, a high voltage system was developed that produces 250,000 beads/min with a diameter of 160 microm +/- 3-5%. The diameter of the beads could be varied between 160-700 microm depending on the needle diameter and construction, the voltage, the distance between the electrodes and the flow of alginate solution. Ca-alginate beads with diameters of 200 and 500 microm were produced by the high voltage electrostatic system. The 200 microm beads were sensitive to poly-L-lysine (PLL) exposure and had to be washed in ion-free solution to avoid collapse. The 200 microm beads swelled more than the 500 microm beads in the washing and PLL treatment. Also, the porosity of the capsules changed with size, but capsules impermeable to tumour necrosis factor (TNF) could be made by exchanging PLL with poly-D-lysine (PDL) for the 500 microm beads. The 200 microm beads were impermeable to IgG after PLL exposure. Islets of Langerhans were encapsulated in alginate-PLL-alginate capsules and evaluated by measuring protruding islets and insulin production. Islets in microcapsules made by the high voltage electrostatic system did not function differently from islets in larger microcapsules made by an air jet system. In conclusion, alginate capsules made by a high voltage electrostatic system enable large-scale production of small capsules with a narrow size distribution that can meet the functional properties of larger capsules by small changes in the encapsulation procedure.

Alginates↗

PubMedDate not supplied

Monocytes stimulated with group B streptococci or interferons release tumour necrosis factor-related apoptosis-inducing ligand.

Tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a cytotoxic member of the TNF family. Some reports have shown that TRAIL is released from cells in a soluble form. In this work, we have investigated the mechanism of release of TRAIL from monocytes. First, we show that whole gram-positive, gram-negative and mycoplasmal bacteria as well as lipopolysaccharide (LPS), interferon-alpha (IFN-alpha), -beta and -gamma all induced upregulation of TRAIL on the surface of human monocytes. Next, we show that IFN-alpha, -beta and -gamma all induced a dose-dependent release of TRAIL, giving significant amounts of soluble TRAIL after 2 days. Of the bacteria, only the Group B streptococcus COH-1 (GBS) induced release of TRAIL and concomittantly induced IFN-alpha. Monocytes stimulated with GBS or IFN-alpha also showed extensive cell death. When monocyte apoptosis was prevented by interleukin-1, GM-CSF, LPS or the caspase inhibitor zVADfmk, the IFN-alpha-induced release of TRAIL was reduced, whereas agents inducing necrosis caused increased release of TRAIL. LPS also prevented release of TRAIL from GBS-stimulated monocytes. The release of TRAIL from IFN-alpha-stimulated monocytes was reduced by inhibitors of both cysteine and metalloproteases. We conclude that bacteria and IFN induce upregulation of membrane TRAIL and that release of TRAIL is associated with cell death.

Amino Acid Chloromethyl Ketones↗

PubMedDate not supplied

Alginate polylysine microcapsules as immune barrier: permeability of cytokines and immunoglobulins over the capsule membrane.

Transplantation of pancreatic islets in alginate polylysine microcapsules is a potential useful method for treating type I diabetes. In this study, the permeability for alginate-polylysine microcapsules to cytokines an immunoglobulines has been investigated by a newly developed method. Magnetic monodisperse polymer particles (Dynabeads) coated with antibodies against selected proteins were encapsulated in 0.7 mm alginate polylysine microcapsules. The capsule membrane permeability to IgG (150 kDa), Transferrin (81 kDa), Tumor necrosis factor (TNF, 51 kDa), Interleukin-1 beta (IL-1 beta, 17.5 kDa), and insulin (5.8 kDa) was estimated by measuring the binding of 125I-labeled proteins to the encapsulated antibody coated Dynabeads. Capsules with an inhomogeneous solid gel core were made of alginates with high guluronic or high mannuronic acid content and poly-L (PLL)- or poly-D-lysine (PDL) of concentrations varied from 0.05-0.2%. The various capsules examined were all impermeable to IgG. The capsules made with a PLL-, but not PDL-membranes were permeable for transferrin. IL-1 beta was found to penetrate all of the different capsule types. The high-G capsules, however, could be made impermeable to TNF and still allowed transferrin to pass. The permeability of these capsules to IL-1 beta, but not to TNF was confirmed in an assay where mouse islets of Langerhans were incubated with TNF and IL-1 beta, and comparing the IL-6 for encapsulated and non-encapsulated islets.

Animals↗