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Biomedical subjects

C F Bassøe

Publications and source records attributed to C F Bassøe.

At least 19 recordsLinked to original sources

Flow cytometric quantitation of human opsonin-dependent phagocytosis and oxidative burst responses to meningococcal antigens.

A one-step flow cytometric (FCM) assay has been developed to quantify both opsonin- and antigen-dependent phagocytosis and intraphagocyte oxidative burst responses. Meningococcal outer membrane structures (OMV) were adsorbed to fluorescent polystyrene beads, opsonized with serum, and exposed to leukocytes. FCM parameters of phagocytosis were evaluated in combinations with oxidative burst indicators. Rhodamine-123 was the most sensitive indicator and was compatible with quantitation of phagocytosis. The phagocytosis and oxidative burst responses induced by OMV beads were dependent on both antigens and opsonins. Increased human opsonic responses against OMV were induced during clinical meningococcal disease. A dissociation was noted between phagocytosis and oxidative burst in individual cells, indicating that functional opsonins against OMV components may differ in their ability to stimulate phagocytosis and oxidative burst responses. The method facilitates evaluation of purified bacterial structures as mediators of opsonin-dependent phagocytosis and intracellular oxidative microbicidal mechanisms, which is of interest in the complex process of selecting bacterial antigens as constituents of certain vaccines.

Adult

Ribosomal proteins sustain morphology, function and phenotype in acute myeloid leukemia blasts.

Translation of mRNA is a prerequisite for cell proliferation, differentiation and viability. We have studied the effect of ribosome protein factors (GPRE) on acute myeloid leukemia (AML) blast cells. Ribosomes were isolated from MPC-11 cells using ultra-centrifugation. GPRE were extracted using a high KCl procedure. Blast cells from six AML patients were grown in suspension cultures for 24 and 96 h. GPRE or granulocyte macrophage-colony stimulating factor (GM-CSF) were added at the start of the incubation. GPRE, but not GM-CSF, prevented chromatin condensation and fragmentation of blast cell nuclei in AML-M2, -M4 and -M5 and the loss of nucleoli in AML-M2 and -M5. The fraction of phagocytosing blast cells in AML-M1, -M2, -M4 and -M5 was increased by GPRE. GPRE stimulated opsonin-dependent and -independent attachment and internalisation of N. meningitidis. GPRE increased the fraction of blasts expressing CD11b and CD32 in AML-M2 and -M5. GPRE diminished the fraction of AML-M5 cells bearing CD35 and CD32. GPRE also decreased the fraction of CD11c-bearing AML-M2 and -M5 cells. GM-CSF potentiated effects of GPRE in AML-M1, -M2, -M4 and -M5. GPRE and GM-CSF in combination affected phagocytosis and surface antigen expression in blast cells that were not influenced by either factor alone. Neither GPRE nor GM-CSF induced terminal differentiation or DNA-synthesis. We conclude that GPRE affects AML blast cell morphology, function and surface molecule expression, possibly by inhibiting apoptosis. The effects of GPRE may be mediated by ribosomal proteins that regulate translation and modulate the subcellular distribution of mRNA species.

Acute Disease

Functional assays for evaluation of serogroup B meningococcal structures as mediators of human opsonophagocytosis.

Functional flow cytometry and chemiluminescence (CL) assays have been modified to identify serogroup B meningococcal structures that mediate anti-meningococcal opsonophagocytosis. Serogroup B meningococcal outer membrane vesicles (OMV) were adsorbed to fluorescent latex beads (OMV-beads) and opsonized with acute phase and convalescence sera from patients with serogroup B meningococcal disease. Phagocytosis of these beads by human monocytes and polymorphonuclear leukocytes (non-lymphocytes) was dependent on both antigen exposure on the bead surface and on serum opsonization. OMV-beads opsonized with serum from a patient recovering from meningococcal disease, caused 97% of the non-lymphocytes to phagocytose an average of 15.8 beads per cell with a CL response of 46,550 mVs, whereas opsonized control beads were phagocytosed by 19% of the non-lymphocytes with 1.1 beads per cell and a CL response of 53 mVs. Increased amounts of functional, anti-OMV opsonins were detected during infection, and opsonized OMV-beads elicited phagocyte responses of similar magnitude to those of opsonized whole meningococci. Phagocyte internalization of OMV-beads was confirmed by confocal laser scanning microscopy. We conclude that epitopes on the meningococcal outer membrane are recognized by anti-meningococcal opsonins in these functional phagocytosis assays, which provide a basis for subsequent evaluation of various purified bacterial components as mediators of human opsonophagocytic responses and hence future vaccine constituents.

Antigens, Bacterial

Effects of interleukin 10 on blast cells derived from patients with acute myelogenous leukemia.

The effect of interleukin 10 (IL-10) on proliferation and cytokine secretion by acute myelogenous leukemia (AML) blast cells was investigated in vitro. IL-10 inhibited spontaneous AML blast proliferation for a majority of patients, whereas in the presence of exogenous growth factors (granulocyte-stimulating factor, G-CSF; granulocyte-macrophage colony-stimulating factor, GM-CSF; interleukin 3) the IL-10 effect on blast proliferation showed a wide variation depending on the individual AML patient. IL-10 seemed to cause an irreversible inhibitory effect on AML blasts, as inhibition could also be demonstrated when IL-10 was present only during the initial preincubation of the leukemia cells. IL-10 also inhibited AML blast colony formation. However, independent of the effect on AML blast proliferation, IL-10 decrease cytokine secretion from AML blast cells for all patients, as demonstrated for IL-1 alpha, IL-1 beta, tumor necrosis factor-alpha, GM-CSF and interleukin 6. IL-10 did not inhibit development of apoptosis in AML blasts cultured in vitro. Expression of complement receptors and capability to adhere and internalize bacteria by AML blasts were not altered by IL-10.

Aged

The skinache syndrome.

Chronic pain of unknown aetiology, and characterized by cutaneous trigger points, has been coined the skinache syndrome. The treatment of the skinache syndrome was evaluated in 94 patients by two independent methods 2 years after treatment. After one subcutaneous injection of lidocaine 68% of the patients were cured. The pain recurred in 27 patients having suffered for an average of 2 years. Surgical removal of the cutaneous trigger points cured 77% of the latter patients. The odds ratio of success of surgical treatment versus all other treatments combined was 101.3. The skinache syndrome requires a precise clinical investigation. Even when the origin of the pain in tendons, muscle and adipose tissue is excluded, the skinache syndrome remains a common, debilitating disorder. In contrast to fibromyalgia, the skinache syndrome has a simple and effective cure.

Aged

[Automatic text analysis of medical records].

Clinical narratives stored in a computerized medical record (PROMED) were automatically analyzed by a computer program (LOGSTORY). No medical knowledge was built into LOGSTORY prior to the analysis. The clinical sample consisted of 5,041 patients and 14,323 diagnoses. The present study concerns 375 diagnoses from general practice and 160 diagnoses from occupational medicine. LOGSTORY reproduced the symptoms, signs, laboratory investigations, anatomy and etiology of diabetes mellitus, obesity and lung diseases. Similarities and differences between the clinical states were automatically recognized and quantified. The extraction of knowledge from the clinical narratives required a problem-oriented medical record. PROMED-LOGSTORY may be useful for self-evaluation, peer review, quality control and research.

Expert Systems

[Standardization of medical forms in health care].

In the Norwegian health care system, resources worth several 100 millions of US dollars are tied up in traditional routines for handling forms. A standard form, combined with computer record systems, could potentially release a considerable share of these resources for alternative uses. Universal Form Medicine (UFM) has been used by Norwegian general practitioners since 1985. The Universal Form Medicine has been a stable standard, and replaces more than 95% of all health forms. Today, about 50% of all doctors with a computerized medical record system can use this standard form. From most aspects, the Universal Form Medicine has advantages over preprinted forms, and can be adapted to silent printers. To our knowledge, no other countries have taken the advantage of standardizing their medical forms.

Family Practice

Semantic analysis of medical records.

A program (LogStory) is described that was developed for the automatic semantic analysis of clinical narratives, stored in a computerized problem-oriented medical record (PROMED). The diagnoses were written in a free-text format during consultation, and later collected into diagnostic classes, e.g., diseases. A lexical parser automatically created dictionaries from the clinical narrative associated with each disease. Automatic (fuzzy) set operations were performed on the words associated with each class. The manifestations of 16 diseases were automatically extracted by pairwise operations on the word sets. The correlation between diseases and corresponding signs, symptoms and treatment was highly significant (p < 0.001). Applying the difference operation on diseases with disjunct sets of clinical findings allowed the recovery of disease-specific knowledge. The evolution of a disease was accounted for, and the system was able to generalize its findings. The PROMED-LogStory concept enables the processing of natural language and may be a powerful tool for knowledge acquisition and clinical research.

Expert Systems

The influence of the host on the course of gastric carcinoma.

Immunoglobulins (Ig) and some complement components (C) were quantified in sera from patients with gastric carcinoma before surgery and at regular intervals during a 5-year follow-up. The preoperative concentrations of C1-INH and C4 were higher (P < 0.0005 and P < 0.005) and IgG lower (P < 0.0005) in 50 patients with recurrence than in 46 5-year survivors. The prognostic significant of C1-INH was superior to that of the extent of disease (F-values 37.1 and 26.1). The preoperative immune data classified 76% of the patients correctly as to recurrence and no recurrence. Also, the preoperative C1-INH concentration had a highly significant effect on time to recurrence of cancer (P = 0.0007), adjusting for age and disease extent. After surgery the mean IgG concentrations were within normal range and without difference between the two groups. On the other hand, the concentrations of C1-INH and C4 in the individual patients in both groups remained the same from before to after surgery and throughout the observation period (P = 0.34). Apparently, the serum levels of C1-INH and C4 do not reflect the bearing of cancer. We therefore suggest that these variables represent an independent immune state that is appropriate to the host. A comparison of our variables with those of healthy individuals seems to support this idea. This immune state has a significant influence on whether a resected gastric cancer will recur, and also on how soon recurrence may be manifest.

Adult

Protein factors in extracts of ribosomes isolated from L-929 cells during various phases of the cell cycle.

Centrifugal elutriation has been utilized in order to separate cultures of L-929 fibroblasts into subpopulations containing cells at different stages of the cell cycle. The subpopulations were characterized by Coulter counter volume determination, [3H]thymidine label into DNA and flow cytometry. When a population of early G1 cells was returned to roller culture it was shown to progress through the cell cycle in a synchronous manner. Ribosomal factor extracts were prepared from cells at various phases during the cell cycle. The amounts of protein in the extracts varied greatly, being lowest in early G1 phase and showing a peak during S phase. Polyacrylamide gel electrophoresis demonstrated that there were differences in the protein species present in the extracts. Some proteins were present in the same amounts throughout the cell cycle, whereas others appeared to show a form of cyclical behaviour.

Animals

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