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C F Bassøe

Publications and source records attributed to C F Bassøe.

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A growth-promoting ribosome extract (GPRE) from mouse L-929 cells stimulates growth of the HL-60 human promyelocytic leukemia cell line.

An extract termed growth-promoting ribosome extract (GPRE), isolated from mouse L-929 cells stimulates growth of HL-60 human promyelocytic leukemia cells. The stimulation first becomes apparent at 72 h when the cells start to enter the quiescent state. The inhibition of protein synthesis by the addition of cycloheximide to L-929 cells before ribosomal extracts were prepared did not alter the stimulatory effect of GPRE. When GPRE was added together with 20% fetal calf serum to cultures of quiescent HL-60 cells, growth was stimulated to the extent that the generation time was reduced by approximately 9 h to 32.4 h. GPRE alone was unable to stimulate the quiescent cells. The growth stimulatory effect was not restricted to one cell generation but was a characteristic of at least the following two cell cycles. GPRE extract from L-cells synchronized by centrifugal elutriation was most efficient when isolated from cells in early G1 phase, while extract from S phase cells had virtually no effect. It is tentatively suggested that the factor belongs to the competence/progression group of growth factors.

Animals↗

Contraception in general practice analyzed by a computerized medical record.

By a computerized medical record 174 patients who consulted a general practitioner regularly for contraceptive advice through 51 months were selected. There were 67 patients aged 25.9 +/- 4.0 (mean +/- SD), using Lynestrenol 0.75 mg and etinyloestradiol 375 micrograms or Levonorgestrel 0.150 mg and etinyloestradiol 30 micrograms. Eleven patients aged 27.2 +/- 5.3 using Noretisteron 0.3 mg and 96 patients aged 33.3 +/- 6.0 using intrauterine contraceptive device (IUCD). As judged by the Pearl index, the Pill-users unwantedly and unexpectedly became pregnant more often than IUCD-users. Acute salpingitis was encountered in six Pill-users, and in one with IUCD. There was more often purulent vaginal discharge and metrorrhagia in the Pill-group, but the hemoglobin values were similar. The importance of these findings remain to be established, but they suggest that the IUCD may be more reliable than the Pill, perhaps due to a better compliance. Eight IUCD patients did not return for changing the device in due time after 36 months of use. Four out of 13 patients with cervical dysplasia did not return for prescribed controls of the cervical smear. The computerized medical record seems to be a powerful tool for the monitoring of high risk patients and clinical research in general practice.

Adolescent↗

Phagocytosis by human leukocytes, phagosomal pH and degradation of seven species of bacteria measured by flow cytometry.

Phagocytosis by human leukocytes, phagosomal pH and degradation of seven species of bacteria were studied by a flow cytometric method. The percentage of phagocytosing leukocytes was similar for all bacterial strains examined, but Salmonella typhi and Neisseria meningitidis were more slowly phagocytosed than other bacteria. The phagosomal pH surrounding the different bacterial species 15 min after the start of phagocytosis were: Streptococcus pneumoniae 4.4; N. meningitidis 4.9; Str. pyogenes 5.1; Staphylococcus aureus 5.2; Escherichia coli 5.3; S. typhi 5.4; and Klebsiella pneumoniae 5.7. For longer incubation periods, the phagosomal pH remained nearly constant. Staph. aureus, E. coli and S. typhi were the most readily degraded of the species tested. The proteins of all bacteria were degraded more rapidly than their DNA as determined by measurements of the loss of fluorescein-isothiocyanate-fluorescence and ethidium bromide-fluorescence, respectively. The rate of degradation varied from one bacterial species to another. The degradation of proteins and DNA was maximal for bacteria residing in a phagosomal environment estimated to be between pH 5.2 and 5.4.

Bacteria↗

Processing of staphylococcus aureus and zymosan particles by human leukocytes measured by flow cytometry.

Staphylococcus aureus were labeled with fluorescein-isothiocyanate (FITC), stained by ethidium bromide (EB), and measured by flow cytometry (FCM). Bacteria were identified by their FITC-fluorescence and discriminated from the leukocyte cell nuclei by the much higher EB fluorescence and lower coefficient of variation of the latter. Following phagocytosis, both the bacterial FITC- and EB-fluorescence decayed. The mean FITC-fluorescence was reduced about 20% after 15 min and 30-50% after 60 min. Zymosan particles were labeled by FITC and incubated with leukocytes for 15 min. Phagocytes were sorted by FCM and the zymosan particles were liberated by sonication. Their forward angle light scatter was reduced by about 50.6 +/- 2.1% and the FITC-fluorescence by 8.7 +/- 1.0% (mean +/- SD). The reduced FITC-fluorescence and light scatter suggests degradation of proteins, and the decay of EB-fluorescence degradation of DNA, but the specificity remains to be established. By this method phagocytes from a patient with systemic lupus erythematosus seemed to have a selective defect in DNA degradation, whereas phagocytes from a patient with acute myelogenous leukemia had a low capacity to degrade bacterial proteins. This technique offers opportunities for automatic measurements of bacteria and zymosan particle degradation by phagocytes.

Female↗

Phagocyte C3-mediated attachment and internalization: flow cytometric studies using a fluorescence quenching technique.

The dynamics of phagocyte C3-mediated attachment and internalization of fluorescein-isothiocyanate (FITC)-labelled zymosan particles was studied by a flow cytometric (FCM) fluorescence quenching technique, using trypan blue as quenching agent. Trypan blue effectively quenched the fluorescence of extracellular, i.e. free and phagocyte-attached, zymosan particles, but did not influence on the fluorescence of particles internalized by phagocytes. During phagocytosis, an average of 2 C3-coated zymosan particles were simultaneously attached to the phagocyte surface, and the number of attached particles could not be increased by increasing the zymosan to leukocyte ratio, the concentration of C3, the incubation time, or by inhibiting internalization by Cytochalasin B. Phagocyte C3-mediated internalization of zymosan particles was dependent on the concentration of complement, and in the presence of sufficient amounts of C3, internalization continued until saturation was reached at 11 particles per phagocyte.

Complement C3↗

Phagocytosis of Staphylococcus aureus by human leukocytes: quantitation by a flow cytometric and a microbiological method.

Phagocytosis of killed, fluorochrome stained or live Staphylococcus aureus by human leukocytes was measured by flow cytometry (FCM) or a microbiological method, respectively. The results were compared to those obtained by simulation using a prey-predator model. In the presence of an initial bacteria-to-phagocyte ratio of 4:1 to 160:1, the percentage of phagocytosing leukocytes was independent of the bacteria and phagocyte concentration. The number of phagocytosed or killed bacteria per phagocyte increased with increasing bacteria and decreasing phagocyte concentration. One per cent pooled human serum was sufficient for maximum phagocytosis to occur, but killing slightly increased in the presence of 10% pooled human serum. With medium or low initial bacteria-to-phagocyte ratios phagocytosis and killing closely corresponded to the results obtained by the prey-predator model. Maximally each phagocyte was associated with 80 bacteria (measured by FCM), about 45 being phagocytosed (internalized) and 40 killed. The model seems suitable for the simulation of phagocytosis and killing of S. aureus by human leukocytes.

Adhesiveness↗

The effect of serum opsonins on the phagocytosis of Staphylococcus aureus and zymosan particles, measured by flow cytometry.

Human leukocyte phagocytosis of S. aureus and zymosan particles was measured by flow cytometry (FCM). Killing of bacteria was measured by a standard microbiological method. When pooled human serum was heated to 56 degrees C for 30 min, the percentage of phagocytosing polymorphonuclear neutrophilic leukocytes (PMNLs), the rate of phagocytosis and killing of S. aureus by the whole leukocyte population were reduced to about 50% of the control values. The results indicate that the impaired phagocytosis and killing were due to the lack of attachment of bacteria opsonized with heat-stabile serum opsonins, mainly IgG, to 50% of the PMNLs. A prey-predator model was used to compare phagocytosis of S. aureus in heated and control serum. The rate of phagocytosis by those PMNLs that were active in heated serum was the same as that of all the leukocytes in control serum, suggesting that heat-labile serum opsonins did not affect the rate of phagocytosis by these PMNLs. PMNLs and monocytes phagocytosed zymosan particles in control serum, but in heated serum only a fraction of the leukocytes, corresponding to the fraction of monocytes phagocytosed. Thus, all PMNLs seem to be capable of phagocytosis by heat-labile serum opsonins. The combined use of S.aureus and zymosan particles may be of advantage in rapid screening of serum opsonin activities and phagocyte function in infectious and haematological disorders.

Adhesiveness↗

Flow cytometric studies on phagocyte function in bacterial infections.

Polymorphonuclear neutrophilic leukocyte (PMNL) phagocytosis of fluorescein-isothiocyanate labelled Staphylococcus aureus was studied by flow cytometry (FCM) in 41 patients with bacterial infections. Practically all phagocytic cells (granulocytes and monocytes) showed active uptake of fluorescent bacteria. However, the mean fluorescence of the phagocytes was about 15% lower than that of cells from healthy volunteers. This reduction was only seen in the male patients, not in the females. The fluorescence reduction was related neither to the type of infectious agent (Gram-positive versus Gram-negative) nor to the white blood cell count or percentage of immature leukocytes. Five patients with cured infections had phagocyte fluorescence similar to that of the controls. In two patients, phagocyte fluorescence was very low, indicating reduced phagocytosis. Phagocyte fluorescence was not related to patient survival.

Adolescent↗

Simultaneous measurement of phagocytosis and phagosomal pH by flow cytometry: role of polymorphonuclear neutrophilic leukocyte granules in phagosome acidification.

Human polymorphonuclear neutrophilic leukocytes (PMNLs) phagocytosed fluorescein-isothiocyanate (FITC)-labelled Staphylococcus aureus. Free bacteria, phagocytes, and nonphagocytes were discriminated and quantified by flow cytometry (FCM). The relative fluorescence of phagocyte-associated and free bacteria (Nf:N) was calculated by dividing the mean phagocyte fluorescence by that of the free bacteria and the number of phagocytosed bacteria. Bactericidal capacity and chemiluminescence were measured by standard methods. The red-to-green fluorescence ratio of acridine orange-stained PMNLs (R/G) was measured by FCM. Degradation of bacteria was monitored by the reduction in FITC and ethidiumbromide fluorescence of bacteria liberated from the phagocytes. Bacterial FITC fluorescence was pH dependent. Nf:N was 0.5 to 0.7. Using a standard curve for the interrelationship between bacterial fluorescence and pH, phagosomal pH was 5.0-5.5. Phagocytes, kept at 4 degrees C for 24 h had Nf:N approximately 1, did not degrade bacteria, but killed them and emitted chemiluminescence. NH4Cl increased phagocyte fluorescence by 27% and decreased R/G by 50%. Cyanide and azide did not affect Nf:N. Nf:N of phagocytes from a patient with chronic granulomatous disease was 32% below, and R/G was 32% higher than the controls. Acidification of the phagosomes seems to be related to discharge of PMNL granule contents and independent of the respiratory burst.

Acridine Orange↗

Phagocytosis of bacteria by human leukocytes measured by flow cytometry.

A new method has been developed for the evaluation of the phagocytic activity of human leukocytes using fluorescently labeled bacteria and flow cytometry. By simultaneous measurement of cellular light scatter and fluorescence, extracellular bacteria, phagocytes, and nonphagocytes could be discriminated and quantified. All leukocytes assumed to be capable of phagocytosis were phagocytosing, and about 90% of these cells were polymorphonuclear neutrophilic granulocytes. Within 15 min 85% of the bacteria were phagocytosed and each phagocyte contained an average of 15-20 bacteria. The phagocytic capacity of the leukocytes from healthy individuals showed minor interindividual and day-to-day variations. This method facilitates a rapid and accurate in vitro evaluation of the phagocytic activity of human leukocytes.

Bacteria↗

Human leukocyte phagocytosis of zymosan particles measured by flow cytometry.

Human leukocyte phagocytosis of fluorescein-isothiocyanate (FITC)-labelled zymosan particles was studied by a flow cytometric (FCM) assay allowing discrimination of adhered and ingested zymosan particles. Free zymosan particles, non-phagocytes and phagocytes could be discriminated and quantified by simultaneous registration of fluorescence and light scatter. All leukocytes capable of phagocytosis were phagocytosing, and within 15 min 80% of the zymosan particles were adhered or ingested. Compared to the FITC-fluorescence of free zymosan particles, the mean fluorescence of phagocyte-associated zymosan particles was reduced by about 35%, indicating ingestion and processing of zymosan particles. Abolishing the FITC-fluorescence of extracellular zymosan particles by crystal violet, the number of zymosan particles adhered and ingested could be calculated from FCM measurements of phagocyte fluorescence. This showed that in 15 min 83% of the phagocyte-associated zymosan particles were actually ingested.

Flow Cytometry↗

[Obesity].

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Female↗

Rieder cells in the cerebrospinal fluid from a patient with viral meningo-encephalomyelitis. A case report.

To the best of our knowledge, the presence of Rieder cells, as a possible expression of a cytopathogenic effect of virus infection in vivo, has not been reported previously. Rieder cells are radially segmented mononuclear cells and were demonstrated in the cerebrospinal fluid (CSF) from a patient with a documented Coxsackie B virus meningo-encephalomyelitis. The Rieder cells were classified enzymatically as promonocytes. DNA-analysis of CSF showed some mitotic activity which, however, might be due to stimulated lymphocytes.

Adult↗

Flow cytometry for the study of phagocyte functions.

Phagocytes play a key role in host defense, and cell defects are associated with increased susceptibility to infection. Flow cytometry offers rapid and reproducible measurements of single cells in suspension and, following staining with one or more fluorochromes, simultaneous examination of several cell functions. Subpopulations of cells can be identified and sorted for morphologic, biochemical, and functional examination, and specially adapted computer systems allow storage of data for subsequent detailed analysis. Several flow-cytometric assays for the study of phagocytes and their interactions with microorganisms have been developed. These assays facilitate the study of (1) phagocyte surface receptors and regulatory molecules; (2) membrane potential; (3) phagocytosis of microorganisms, including the discrimination between attachment to the phagocyte surface and actual internalization; (4) phagosomal pH; (5) degranulation and enzymatic activity; (6) intracellular calcium; (7) oxidative metabolism; (8) intracellular killing of microorganisms; (9) degradation of microorganisms; and (10) exocytosis. In addition, the influence of serum opsonins on phagocyte-microorganism interactions can be studied. Flow-cytometric techniques are applicable to both experimental and clinical work.

Cell Separation↗