Biomedical subjects

R Bjerknes

Publications and source records attributed to R Bjerknes.

At least 19 recordsLinked to original sources

PubMed1992

Colony-forming unit-granulocyte-macrophage and DNA synthesis of human bone marrow are circadian stage-dependent and show covariation.

Bone marrow samples from sternum and iliac crests were harvested every 4 hours during 19 24-hour periods from 16 healthy male volunteers, and myeloid progenitor cells were cultured by the colony-forming unit-granulocyte-macrophage (CFU-GM) assay. A large interindividual variation was observed in the mean number of colonies during each 24-hour period, with the highest 24-hour mean colony number being about 600% greater than the lowest (range: 16 +/- 2.3 to 100.3 +/- 4.5). For each individual the difference between the lowest and highest colony number throughout the day ranged from 47.4% to 256.3% of the mean colony number of each series. A circadian stage-dependent variation in the number of colony-forming units of myeloid progenitor cells (CFU-GM) of human bone marrow was demonstrated, with values 150% higher, on the average, during the day as compared with the night. The overall data (891 CFU-GM replicates) exhibited a significant 24-hour rhythm (P less than .001) with an acrophase at midday (12.09 hours with 95% confidence limits from 10.32 to 13.49 hours) and a trough at midnight. This 24-hour variation was found to covary with DNA synthesis in the total proliferating bone marrow cell population. A seasonal effect on CFU-GM numbers was detected by ANOVA (P = .014) and by the least squares fit of a 1-year cosine (P = .015), with the highest number found in summer. The potential relevance of these findings should be examined in relation to cytotoxic cancer therapy, use of hematopoietic growth factors, and bone marrow transplantation.

Adult↗

PubMed1992

Ganciclovir therapy of congenital cytomegalovirus disease.

A newborn with cytomegalovirus disease with cerebral involvement was treated with ganciclovir for 21 days. The treatment resulted in only temporary cessation of virus shedding in the urine and probably had no ameliorating effect on the long-term clinical outcome.

Cytomegalovirus Infections↗

PubMed1992

Cell kinetics of the rat corneal epithelium.

In the rat corneal epithelium the mitotic rate (MR) is almost equal throughout the epithelium in the morning (Haaskjold et al. 1988). The labelling index (LI) shows a marked reduction in the central cornea, which could suggest a lack of uptake of tritiated thymidine via the salvage pathway (Haaskjold et al. 1989). In the present study we have used [3H]deoxycytidine, and [3H]thymidine after prior treatment with a methotrexate regimen to elucidate this discrepancy. Deoxycytidine is incorporated into DNA independent of thymidine kinase, while methotrexate, which depletes the cells of reduced folates, makes the cells completely dependent on the salvage pathway. With both techniques the same pattern of labelling was observed, confirming that in the morning the ratio between the MR and the LI differs throughout the cornea. Based on previous observations, an analysis of the MR/LI ratio during 24 h was performed, showing that these parameters were strongly correlated. This suggests that there may be different circadian variations in the cell proliferation parameters throughout the corneal epithelium. The methotrexate regimen may be a useful tool to investigate the salvage pathway.

Animals↗

PubMed1992

Cross-reacting serum opsonins in patients with meningococcal disease.

We have examined the opsonic activity of sera from patients with Neisseria meningitidis (B:15:P1.16) infections against different meningococcal strains, using flow cytometry and luminol-enhanced chemiluminescence. A marked increase in the phagocytosis of ethanol-fixed meningococcal strains of different serogroups, serotypes, and serosubtypes was demonstrated in the presence of convalescence sera compared with acute sera. Convalescence sera also caused a significant increase of leukocyte oxidative metabolism during phagocytosis, as measured by luminol-enhanced chemiluminescence. The sera contained a broad range of opsonins cross-reacting with serogroup A, B, C, W-135, and Y meningococci of different serotypes and serosubtypes, indicating that the cross-reacting opsonins recognized surface epitopes other than those determined by current serotyping schemes.

Adolescent↗

PubMed1992

The early cell kinetic response during healing of corneal epithelial wounds.

The labeling index and the mitotic rate were measured at 4-hour intervals during the first 24 hours after a central abrasion had been made in the corneal epithelium of six groups of four rats each. The proliferative response was noted in the conjunctival, the limbal, and the corneal epithelium. After 24 hours, the density of epithelial cells was equal throughout the corneal epithelium, but there was only half the normal number of cells. Physiological mechanisms seem strongly to regulate the total number of cells per square unit throughout the healing corneal epithelium, but the nature of these mechanisms is unknown.

Animals↗

PubMed1991

DNA synthesis in human bone marrow is circadian stage dependent.

Fraction of human bone marrow (BM) cells in DNA synthesis has been studied by sampling BM from the sternum or the iliac crests every 4 hours during one 24-hour period in 16 healthy male volunteers. Three of the subjects underwent the sampling procedure twice, resulting in 19 24-hour profiles. The percentage of cells in DNA synthesis measured by flow cytometry demonstrated a large variation along the circadian time scale for each 24-hour profile, with a range of variation from 29% to 339% from lowest to highest value. Seventeen profiles (89.5%) had the highest DNA synthesis during waking hours between 08:00 hours and 20:00 hours, and the lowest percentage of cells in DNA synthesis between 00:00 hours and 04:00 hours. The mean value of the lowest DNA synthesis for each 19 24-hour period was 8.7% +/- 0.6%, while the mean value of the highest DNA synthesis was 17.6% +/- 0.6%, ie, a twofold difference. There was no difference in DNA synthesis between winter and summer. A significantly higher DNA synthesis was demonstrated for samples obtained from sternum as compared with the iliac crests, but the same circadian pattern was demonstrated for both localizations. By taking circadian stage-dependent variations in DNA synthesis into account it may be possible to reduce BM sensitivity to cytotoxic chemotherapy, to increase the effect of hematopoietic growth factors as well as increase the fraction of proliferating cells with careful selection of time of day for harvesting BM cells for auto- or allografting.

Adult↗

PubMed1991

Circadian variation in cell proliferation and maturation. A hypothesis for the growth regulation of the rat corneal epithelium.

The rat corneal epithelium has been chosen as a model for studying growth regulation. In this epithelium a large single cohort of cells enters the S phase during a fairly short time period once a day. The factor responsible for this wave of cell proliferation is unknown, but it may be a chemical signal from the central nervous system (the suprachiasmatic nucleus or the corpus pineale). The mature cell compartment of the corneal epithelium is assumed to produce a negative feedback factor (chalone), counteracting the effect of the circadian proliferative factor on the local cell proliferation. When no circadian factor is being produced, during most of the 24 h, the chalone seems to enhance the maturation process. During diminished chalone production (e.g. after cell injury and subsequent regeneration), we will get a more or less unrestricted cell proliferation in the tissue with a delayed maturation process prolonging the chalone depletion. This interaction between the circadian proliferative factor and the negative feedback factor for regulation of proliferation with its accompanying stimulatory effect on maturation, may represent a general mechanism in the regulation of cell proliferation in any tissue. Since in at least some organs virtually all cells entering the S phase do this as a single wave once a day, this mechanism may be enough to explain the regulation of cell proliferation during both normal and regenerative conditions.

Animals↗

PubMed1991

The IgG FcRII and the PI-linked IgG FcRIII trigger cytoplasmic calcium fluxes independently in human granulocytes.

The aim of this study was to investigate signal transduction through the two Fc receptors for IgG on human granulocytes. Using flow cytometry and the calcium indicator Fluo-3, we measured changes in leucocyte cytoplasmic calcium concentrations following cross-linking of cellular Fc receptors with specific antibodies. Two different approaches were used in order to study the two Fc receptors independently of each other. One was to avoid the presence of IgG Fc fragments, capable of binding to both types of receptors. The other was to use leucocytes from a patient with paroxysmal nocturnal haemoglobinuria (PNH) deficient in granulocyte FcRIII. In contrast to earlier reports, both approaches showed that the two types of IgG Fc receptors on granulocytes are capable of increasing cytoplasmic free calcium concentrations independently of each other. The results suggest that free cytoplasmic calcium ions are involved in the signal transduction pathway of both types of IgG Fc receptors on human granulocytes.

Adult↗

PubMed1991

Regeneration of the rat corneal epithelium after injury. A cell kinetic study comparing the reaction after abrasions made mechanically and with n-heptanol.

Central corneal epithelial defects with diameter 3.5 mm were made with n-heptanol in the right eyes of rats. Erosions of the same size were made by mechanical scraping in the left eyes of the same animals. All the erosions were covered by epithelium after one day. After one day the mitotic rate and the labelling index were higher in the n-heptanol treated corneas compared to the mechanically abraded ones. Both methods produced similar mitotic rates and labelling indexes after 3 and 12 days. The results from wound healing studies using n-heptanol or mechanical scraping are therefore not entirely comparable because of the different regenerative responses after one day. However, the n-heptanol method is easier to perform and to standardize, and is therefore preferable.

Alcohols↗

PubMed1990

Signal transduction in human monocytes and granulocytes through the PI-linked antigen CD14.

A possible role for the PI-linked CD14 molecule in human monocyte and granulocyte signal mediation was investigated. Using flow cytometry and the fluorescent indicators Fluo-3 and dihydrorhodamine-123 it was shown that crosslinking of the CD14 molecule induces an increase in monocyte and granulocyte cytoplasmic calcium concentration and monocyte H2O2 production. These responses were found to be independent of IgG Fc receptors and suggest an intrinsic signal mediating capacity of the CD14 molecule.

Antigens, CD↗

PubMed1990

Flow cytometric assay for the measurement of human bone marrow phenotype, function and cell cycle.

A flow cytometric assay for the measurement of human bone marrow and blood leukocyte antigen expression, phagocytosis, and proliferation is described. Subpopulations of leukocytes were identified by their light scatter characteristics, and the expression of a myeloid differentiation antigen (designated CDw65) determined following incubation with CDw65 specific fluorescein-isothiocyanate (FITC) conjugated monoclonal antibodies (VIM2). Incubation of leukocytes with ethidium monoazide (EMA) labeled Candida albicans followed by staining with FITC conjugated VIM2 allowed the combined determination of cellular CDw65 expression and phagocytic capacity. In addition, immunostained leukocytes were fixed, and their DNA labeled with propidium iodide (PI), before CDw65 expression was measured for cells in different phases of the cell cycle. The method allows evaluation of phenotypic and functional heterogeneity, as well as cell cycle parameters, within subpopulations of cells during hematopoietic differentiation.

Adult↗

PubMed1990

Circadian variations in the DNA synthesis of the rat corneal epithelium. A study using double labelling with tritiated thymidine.

A prominent circadian rhythm was found in the labelling indices (LI) of the peripheral rat corneal epithelium and of the adjacent conjunctival epithelium, while almost no diurnal variation was found in the central area. Application of a double labelling technique indicated that there are rhythmic pulses of high and low influx of cells into the S phase and similar pulses of efflux of cells from the S phase. Results of the study indicate that there are different cohorts of cycling cells all over the rat corneal epithelium. Cells belonging to a rapidly proliferating cohort are observed in the peripheral cornea. There is a gradual reduction in the fraction of labelled DNA-synthesizing cells towards the centre. The considerably lower fraction of cells taking up tritiated thymidine (3H)TdR in the central cornea may be due to a higher fraction of basal cells having reached higher levels of differentiation. This may result in a shift from the salvage to the de novo pathway. The slowly proliferating cohort seems to have a prolonged S phase duration and displays practically no diurnal variation in the LI. The DNA-synthesizing cells belonging to this latter cohort probably use the salvage pathway for DNA synthesis resulting in uptake of (3H)TdR all over the cornea. The LI is thus not a reliable indicator of cell proliferation in the corneal epithelium, due both to the heterogeneity of the cell proliferation, and in particular due to the lack of labelling of the centrally located DNA-synthesizing cells. To what extent these properties may also be present in other proliferating tissues with different levels of differentiations, may be questioned.

Animals↗

PubMed1990

Two cell kinetic methods studied on the rat corneal epithelium.

A stathmokinetic method (using Colcemid) and the [3H]thymidine technique (pulse labelling with tritiated thymidine, [3H]TdR) have been evaluated in the rat corneal epithelium. The dose is not of critical importance for the Colcemid method, thus indicating an all or nothing effect within the dose range studied. A one point estimate is sufficient to calculate the mitotic rate (MR), and in the rat corneal epithelium a 4 h accumulation period is recommended. After administration of [3H]TdR there is an increasing response with increasing dose, followed by a levelling off at higher doses. It seems reasonable to use the lowest maximal effective dose. The labelling index (LI) can be reliably registered 1 h after administration of the drug. For each of the drugs we found corresponding results after topical application and intraperitoneal injection. Hence, topical application of small doses of both Colcemid and [3H]TdR makes interesting in vivo experiments on larger animals and even on human beings possible. Due to the extreme regularity of the corneal epithelium this part of the eye is an interesting organ for cell kinetic studies and provides an excellent tool for evaluating cell kinetic methods.

Administration, Topical↗

PubMed1990

Altered neutrophil functions in patients with large burns.

Neutrophil functions were examined longitudinally in 16 patients with large burns using multiparameter flow cytometry (FCM). At admission and through the first 10 days, the neutrophil expression of surface complement receptors for C3b (CR1) and C3bi (CR3) was increased, whereas neutrophil Fc receptor III (FcRIII) expression decreased. The phagocytosis of C3-opsonized Candida albicans increased during the same time period, whereas the ingestion of Ig-opsonized C. albicans decreased. The neutrophil intracellular killing of C. albicans was reduced by about 25% at admission. The microbicidal capacity was further compromised during the next 2 weeks, with a 50% reduction of intracellular killing 10 days following injury. The kinetics of neutrophil phagolysosomal acidification was altered during the first 20 days after burn injury, as the initial alkalinization of the phagolysosomes documented in control neutrophils could not be demonstrated in patients cells. In addition, patient neutrophil H2O2-production, which was only slightly reduced at admission, was gradually decreased during the first 2 weeks, with an oxidative burst about 40% lower than controls at day 10. All neutrophil functions tested were normal at discharge. The results demonstrate significant metabolic and functional alterations in neutrophils from patients with large burns. The data are consistent with a general activation of circulating neutrophils in the early phase after thermal injury, which is followed by impairment of neutrophil microbicidal mechanisms that may predispose for infectious complications.

Adult↗

PubMed1990

Impairment of rat polymorphonuclear neutrophilic granulocyte phagocytosis following repeated hypobaric hypoxia.

Exposure of rats to repeated hypobaric hypoxia (17 h at 0.5 atm for 0-7 d) induced significant reduction of the phagocytic capacity of peripheral blood polymorphonuclear neutrophilic granulocytes (PMNL). In addition, the hypoxia induced a significant increase of the hematocrit, hemoglobin, thrombocyte, and total leukocyte concentrations. Differential counting of peripheral blood revealed significant granulocytosis. An increase of the concentration of corticosterone was demonstrated following 2 d of hypobaric hypoxia, whereas 5 d later the corticosterone concentrations were similar to that of the controls. The reduced PMNL phagocytic capacity observed following repeated hypobaric hypoxia may have consequences for host defence in situations of hypoxia exposure.

Aerospace Medicine↗

PubMed1989

Flow cytometric assay for the measurement of serum opsonins to Neisseria meningitidis serogroup B, serotype 15.

A flow cytometric phagocytosis assay has been developed for the measurement of human serum opsonins to serogroup B meningococci. Live bacteria and bacteria inactivated by heat, formalin or ethanol were labelled with fluorescein-isothiocyanate (FITC). The bacteria were opsonized with sera from patients with group B meningococcal disease and sera from healthy controls, and phagocytosis determined by combined measurements of FITC-fluorescence and forward angle light scatter. Optimal sensitivity was obtained using viable bacteria, 5% serum, 20 bacteria per leukocyte capable of phagocytosis, 7.5 min opsonization time, 5 min phagocytosis time, 37 degrees C, and continuous agitation during opsonization and phagocytosis. The opsonic activity of sera from convalescent patients was markedly higher than that of sera from patients with acute illness. Only minor day-to-day and interindividual variations were observed. The flow cytometric phagocytosis technique is a rapid and reproducible method for the measurement of serum opsonins to meningococci.

Blood Bactericidal Activity↗

PubMed1989

Circadian variation in the mitotic rate of the rat corneal epithelium. Cell divisions and migration are analyzed by a mathematical model.

A stathmokinetic method was used to study the diurnal variation in the mitotic rate (MR) of the rat corneal epithelium, and in the adjacent conjunctival epithelium. A prominent circadian variation in cell proliferation was observed in both epithelia, both showing almost the same pattern, which may indicate that both tissues are submitted to the same regulatory mechanisms. The average rate of cell renewal during a 24 h period indicated a mean cell renewal time of 12.3 days. This is longer than previously assumed. The MR declined toward the central cornea. Based on the above observations and the known centripetal migration of cells in the corneal epithelium, we have developed a mathematical model showing isomorphism with the renewal of the corneal epithelium.

Animals↗

PubMed1989

Neutrophil dysfunction after thermal injury: alteration of phagolysosomal acidification in patients with large burns.

The neutrophil phagolysosomal acidification during phagocytosis of Staphylococcus aureus was examined in six patients with large burns, using a flow cytometric technique allowing the simultaneous measurement of phagocytosis and phagolysosomal pH. The kinetics of neutrophil phagolysosomal acidification were altered during the first 20 days following injury, as the initial alkalinization of the phagolysosomes documented in control neutrophils could not be demonstrated in patient cells. Only at discharge and follow-up were the kinetics of phagolysosomal acidification normal. In addition, measurements of neutrophil maximal phagolysosomal acidification showed a lower pH in patient phagolysosomes than in the controls during the first 5 days of hospitalization. The changes of phagolysosomal acidification did not correlate with the alterations of neutrophil maturity or phagocytic capacity. The results demonstrate alterations of an oxygen-independent microbicidal mechanism in neutrophils from patients with large burns, which may contribute to the reduced capacity of neutrophil intracellular killing following thermal injury.

Adolescent↗