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Biomedical subjects

T Kouri

Publications and source records attributed to T Kouri.

At least 19 recordsLinked to original sources

Estimation of reference change limits using patient data.

Two approaches for deriving reference change limits from patient data are described. In the direct method, hospital database information is used for the selection of appropriate reference groups. If database information is not sufficient or reliable enough, but still most of the source data can be considered as health-related, an indirect method can be applied in the calculation of rough estimates for reference change limits. A computer program developed by us, GraphROC for Windows, includes both methods for the estimation of change limits from patient data. Time between specimen collections should be included as one classifying factor in the selection of source data. When only one previous result is available for comparison, change limits based on the reference sample group form the only available guide for clinical interpretation. However, when several previous results are available and the within-subject variances for the considered analyte are known to be heterogeneous between individuals, the clinical interpretation should rather be based on application of time series analysis.

Data Interpretation, Statistical

Reliability and adequacy of discharge diagnosis databases in the production of reference values.

Discharge diagnoses provide a possibility to select patients individually and then to establish reference values for both "pathological" and control groups. Currently, the available diagnostic information is still at its infancy and should be carefully evaluated before the reference values based on those groups are utilized. It is anticipated that electronic storage of diagnostic and therapeutic information will be applied more commonly in the future as the development of computers makes it easier. The advanced utilization of laboratory data challenges physicians both in the clinical and laboratory side to participate in this development in order to make the information systems serve their actual needs more closely.

Clinical Laboratory Information Systems

Reference intervals developed from data for hospitalized patients: computerized method based on combination of laboratory and diagnostic data.

We utilized the databases of a hospital information system to select for determination of reference values various individual hospitalized patients on the basis of their diagnoses at discharge. The nonparametric 2.5-97.5% "health-related" reference intervals were calculated for hemoglobin concentration, mean corpuscular volume (MCV), and erythrocyte count for both sexes. After excluding patients with diseases possibly affecting erythrocyte variables, we obtained a final group of 1786 women and 1450 men, ages 20-65 years, who were studied in age groups of 20-30, 30-45, 45-55, and 55-65 years. The upper reference limits of the MCV results obtained from hospitalized patients were higher than those produced conventionally from healthy individuals, as would be intuitively suggested by clinical experience. This method, based on selection by diagnosis, could be applicable to various analytes measured in hospital laboratories, provided sufficient data are available as databases.

Adult

Performance of NycoCard:::U-Albumin and Micral-Test rapid methods for detecting microalbuminuria.

NycoCard:::U-Albumin and Micral-Test semiquantitative methods were evaluated against nephelometric measurements of nightly albumin excretion rates from 159 consecutive diabetic patients. Both methods can be safely applied for screening of intact renal function even at a strict prediction limit for incipient nephropathy of albumin excretion rate = 15 micrograms/min (standardised specimens collected at bed-rest), since the predictive values of negative tests were 95-100% for both tests. Predictive values of positive tests were 38% (NycoCard:::U-Albumin) and 44% (Micral-Test), if the designated value of 10 mg/l was used as the limit of positivity. By using 20 mg/l as the limit, the predictive values of positive tests were 62% (NycoCard:::U-Albumin) and 72% (Micral-Test), indicating the need for quantitative measurements in patients with positive screening results at both these limits.

Albuminuria

Quality control of multichannel hematology analyzers. Bivariate on-line comparison of MCV and MCH values for the detection of random errors.

Volume and hemoglobin content of red blood cells are physiologically interdependent variables but are measured independently in multichannel hematologic analyzers. Therefore correlation of these results to each other can be used as an additional step in internal quality control. To facilitate precise correlation, data were collected from 44,873 consecutive routine determinations of red blood cell parameters. Gaussian distributions were fitted to the original distributions and bivariate elliptical acceptance limits were calculated for simultaneous evaluation of mean corpuscular volume and mean corpuscular hemoglobin results. With the proposed method, rare combinations of the three directly measured red blood cell parameters (hemoglobin, red blood cell count, and mean corpuscular volume) can be identified for the detection of random analytical or preanalytical errors. This mean corpuscular volume-mean corpuscular hemoglobin rule could be added to internal quality-control programs of hematologic analyzers to improve detection of unusual results and possible random errors.

Diagnosis, Computer-Assisted

Occurrence of two germline-related rheumatoid factor idiotypes in rheumatoid arthritis and in non-rheumatoid seropositive individuals.

Human rheumatoid factor (RF) paraproteins express two distinct light chain cross-reactive idiotypes defined by the monoclonal antibodies 17.109 and 6B6.6. These germline gene-related cross-reactive idiotypes are both carried on VK3 light chains and are each present on about one-third of IgM RF paraproteins. We assessed the degree to which these idiotypes are represented in polyclonal RFs. We used rheumatoid arthritis (RA) and non-RA RF-positive sera selected from a large cross-sectional population study (the Mini-Finland Health Survey), and sera from a community-based follow-up study of recent-onset RA patients from Heinola, Finland. In the Mini-Finland Health Survey, elevated levels of the 17.109 RF idiotype were seen in sera of 13% of the RA and 19% of the non-RA group; 6B6.6 RF was seen in 26% of the RA and 28% of the non-RA group. In sera of the Heinola follow-up study, 17.109 RF was seen in 12% initially, but in only 3% at 8 years. Similarly, 6B6.6 RF was detected in 25% initially, but in only 7% at 8 years. Ten sera positive for RF prior to the onset of clinical RA were identified from individuals of a second large population study from Finland (North Karelia project); two of these sera exhibited the 6B6.6 idiotype; none exhibited the 17.109 idiotype. The data are consistent with the concept that these germline gene-related cross-reactive RF idiotypes occur frequently in the polyclonal RF of non-RA as well as RA sera, and that in RA the idiotypes may sometimes be reduced or lost as a consequence of somatic diversification of the RF through somatic mutation, usage of new germline genes, or both.

Arthritis, Rheumatoid

Antibodies to synthetic peptides from Epstein-Barr nuclear antigen-1 in sera of patients with early rheumatoid arthritis and in preillness sera.

We studied IgG antibody levels to synthetic peptides (p62, E11 and E3) from Epstein-Barr nuclear antigen-1 (EBNA-1) protein sequence in sera of patients with rheumatoid arthritis (RA), in pre-RA and in rheumatoid factor (RF) positive non-RA sera from Finland. Anti-E11 and anti-E3 antibody levels were significantly elevated in RA sera, compared with both RF negative and RF positive nonrheumatoid controls. The concentrations of anti-E11 and anti-E3 antibodies in the preillness sera were lower than those in RA sera. Eight-year followup samples of patients with RA had slightly decreased levels of anti-E3 and anti-E11 antibodies compared with samples taken within 6 months of the disease onset. Anti-p62 antibody levels did not differ significantly between any of the groups studied. Thus, elevated levels of antibodies to 2 of the glycine containing EBNA-1 peptides were associated with clinical RA.

Adult

DNA synthesis in prolyl 4-hydroxylase positive fibroblasts in situ in synovial tissue. An autoradiography-immunoperoxidase double labeling study.

DNA synthesis in prolyl 4-hydroxylase (PH; EC 1.14.11.2) positive fibroblasts in situ in synovial tissue was studied using an autoradiography-avidin-biotin-peroxidase complex (ABC) double labeling. Fibroblasts in monolayer culture and in situ in synovial tissue were PH positive, whereas freshly isolated peripheral blood lymphocytes, monocytes, dendritic cells and granulocytes were PH negative. In rheumatoid arthritis (RA) 37 +/- 3 (22-56)% of all DNA synthesizing cells in situ were PH containing fibroblasts, whereas all DNA synthesizing cells in patients with meniscus lesion were PH positive. In both conditions, more than half of the self-replicating fibroblasts were located in the lining cell layer. This is probably not an artifact caused by insufficient penetration of 3H-thymidine because most of the DNA synthesizing lymphocytes were deep down in the synovial stroma. In RA 51 +/- 8 (17-88) PH positive fibroblasts in the S phase of the cell cycle were observed/3 mm2 synovial tissue, whereas the corresponding figure in meniscus patients was only 1 +/- 1 (0-5) (p less than 0.01). This suggests that the local fibroblasts in RA are activated, probably as a result of various fibroblast growth factors produced locally as a result of the inflammatory synovitis. In RA however, less than 1% of all local fibroblasts were self-replicating in situ, whereas labeling indices over 5% were not uncommon in RA synovial fibroblast cultures. This finding suggests that uncontrolled fibroblast proliferation is regulated in vivo by negative feedback mechanisms.

Arthritis, Rheumatoid

Activated T lymphocytes in patients with multiple sclerosis in clinical remission.

Activation state and proliferation of lymphocytes of 6 patients with definite multiple sclerosis in clinical remission were studied. Lymphocytes carrying MHC class II coded Ia antigen, glycoprotein gp 40/80 and interleukin-2 receptor were significantly higher in patients with MS in remission than in normal healthy controls. The entrance of T cells into the G1 and S phase of the cell cycle was studied using [3H]thymidine autoradiography in combination with immunocytochemistry (cell markers). The proportion of T cells in the S phase of the cell cycle in the peripheral blood was similar in the MS patient and in the control group.

Autoradiography

Adherent cells from rheumatoid synovia: identity of HLA-DR positive stellate cells.

Rheumatoid synovia were enzymatically digested and the in vitro morphology of different types of plastic adherent cells was observed. Four main types of cells were found after 24 hours in culture: stellate cells which had nuclei resembling those of classical cultured fibroblasts, but which stained positively with I2 antibody (anti-HLA-DR antibody); fibroblastic cells; cells which resembled morphologically in vitro macrophages and which were I2 and OKM-1 positive; round monocytes. The stellate cells did not stain with anti-S-100 or OKT-6 antibodies, which are used to detect classical antigen presenting dendritic cells. Furthermore, in the presence of indomethacin the stellate shaped cells were replaced by new I2 positive cells with a typical fibroblast shape. These results support the view that the stellate cells in synovial cell cultures represent HLA-DR positive fibroblasts, probably B cells of synovial lining.

Arthritis, Rheumatoid

Etiology of rheumatoid arthritis.

Definite genetic associations with immunological cooperative HLA-D(R) antigens have been demonstrated for rheumatoid arthritis (RA). Microbial etiology has not been proven, but some hope for the supporters of this view is still given by small viruses, plasmids of enteric bacteria or perhaps oncogen-like DNA-sequences. Yet, electrophoretical analysis of membrane proteins or surface glycoproteins of RA synovial cells does not show any differences compared to reference cells. Autoimmunity to several tissue elements has been demonstrated, but most of it is of secondary nature. Antigenicities of type II and III collagens are probably only contributory factors for HLA-DR4 positive individuals. Proteoglycans or minor cartilage collagens have not been extensively studied, so far. Endocrine, dietary or psychological influences might be triggering events for otherwise 'preloaded' individuals.

Animals

Neurofibromatosis tumor and skin cells in culture. II. Structural proteins with special reference to the cytoskeletal and cell surface components.

Structural proteins of cultured neurofibromatosis (NF) tumor and skin cells were studied with reference to control skin fibroblasts. In polyacrylamide gel electrophoresis (PAGE)/fluorography the banding patterns of the cell lysates were markedly similar. NF tumor cells, however, produced a 60 kD band with a stronger and a 48 kD band with a lighter protein staining and metabolic labeling intensity. Furthermore, skin cells were also characterized by a 26 kD protein and the tumor cells by a 22 kD protein with high metabolic labeling intensity. Neuraminidase/galactose oxidase/NaB3H4-labeled NF skin and control skin cells possessed a 220 kD protein that was less intensively labeled in the tumor cells. The banding pattern of the skin cells was also characterized by a protein with slightly lower molecular weight (86 kD) than that of the tumor cell lysates (90 kD). In all cell lines studied indirect immunofluorescence stainings revealed bright arrays of vimentin type intermediary filaments but no desmin, cytokeratin, glial fibrillary acidic protein (GFAP), or neurofilament proteins. NF skin and control skin cells possessed well developed actin-containing bundles of microfilaments, while those of the tumor cells lacked a typical stress-fiber organization. The general morphology of the tumor cell cultures was also irregular. Transmission electron microscopy revealed no basic differences in the structure of intermediary filaments or microfilaments. The present data provide basic knowledge of neurofibromatosis skin and tumor cells and demonstrate that cultured cells originating from neurofibromas are defective in both their intracellular and extracellular organization.

Actins

Antigenicity of proteins from cultured synovial fibroblasts.

We were able to characterize about 25 antigenic proteins of cultured synovial fibroblasts with apparent molecular weights of 15-230 kd by SDS-electrophoresis and immunoblotting using guinea pig antiserum. Some of these proteins were bovine-serum-derived, adsorbed onto fibroblasts from culture medium. Both rheumatoid and normal sera contained natural antibodies reacting with synovial fibroblast as well as bovine serum antigens when studied with immunoblotting. The amounts of antibodies were quantitated with enzyme immunoassay, measuring IgG and IgA class antibodies against plasma membrane preparations of rheumatoid and normal synovial fibroblasts. No rheumatoid arthritis-related changes were detected in synovial fibroblast protein antigens or antibodies against them in rheumatoid patient specimens.

Antibodies

Surface membrane glycoproteins of macrophage-like synovial cells from rheumatoid and control patients.

Synovial cells were prepared by enzyme digestion and Percoll gradient centrifugation of rheumatoid arthritis (RA) synovial specimens or by trypsin-rinsing of non-inflammatory cadaver joints. Most (70-80%) of the cells from RA patients were OKIa -positive macrophage-like cells, 10-20% other OKIa -positive cells, and about 10% fibroblastic cells, whereas 90% of the normal synovial cells were OKIa -positive macrophage-like cells and the rest fibroblasts. These adherent synovial cells were compared with fibroblastic synovial cells obtained by sequential passaging of explanted dividing cells. Periodate-[3H]borohydride labelling followed by SDS-gradient gel electrophoresis demonstrated similar sialo-glycoprotein patterns in both the adherent synovial cells and synovial fibroblasts. The molecular weights of the main surface glycoproteins resembled closely those of skin fibroblasts but not those of peripheral blood monocytes. RA samples showed inconsistent heterogeneity. The results indicate either that all synovial cells possess a similar basic structure or that macrophages of peripheral blood origin express fibroblastic contact glycoproteins when settling down into synovium.

Arthritis, Rheumatoid

Morphologic alterations in cultured human synovial fibroblasts induced by blood mononuclear cells.

Effects of peripheral blood mononuclear cells on cultured synovial fibroblasts were studied. When mononuclear cells from normal or rheumatoid blood were incubated on synovial fibroblast cultures, a part of the cells adhered to the fibroblasts. They were mainly T lymphocytes but also some B lymphocytes and monocytes. After a 10-hour incubation, adhered mononuclear cells induced morphologic alterations to synovial fibroblasts: appearance of stellate cells and thinning and branching of fibroblasts. No changes were seen when the cells were incubated in the presence of indomethacin. Cytotoxicity of peripheral blood mononuclear cells from 8 rheumatoid patients was also tested against three rheumatoid and three normal synovial fibroblast strains. Only 2 out of 48 combinations were cytotoxicity. The potentially cytotoxic mononuclear cells were bound equally well to rheumatoid and control synovial fibroblast cultures.

Arthritis, Rheumatoid

Characterization of plasma membranes and rough endoplasmic reticulum of synovial cells cultured from rheumatoid arthritis patients.

[35S]methionine-labelled plasma membrane (PM) and rough endoplasmic reticulum (RER) proteins were analysed from synovial cell cultures derived from rheumatoid arthritis (RA) and reference patients. Pure RER was prepared by a modification of CsCl-containing sucrose-cushion method. The PM proteins were resolved into 30-35 bands using sodium dodecyl sulphate-electrophoresis in polyacrylamide gradient gels. The major polypeptides had apparent molecular masses of 170, 140, 110, 60, 40, 33, 23 and 14 kilodaltons. The major RER polypeptides had the apparent molecular masses of 170, 74, 58 and 35 kD. No reproducible differences were demonstrated in the polypeptide patterns or in the specific activities of the PM and ER (endoplasmic reticulum) marker enzymes between cells from RA and reference patients. The results reflect similarities of membrane structures in RA and reference cells, although differences have been reported in their metabolism.

Arthritis, Rheumatoid