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Biomedical subjects

M Kameko

Publications and source records attributed to M Kameko.

At least 19 recordsLinked to original sources

Inflammatory cytokine cascade released by leukocytes in cerebrospinal fluid after subarachnoid hemorrhage.

Subarachnoid hemorrhage (SAH) elicits an inflammatory response in the subarachnoid space, which is mediated by the release of various cytokines. To assess their involvement in post-hemorrhagic complications, we determined the source and time-course of the release of inflammatory cytokines and acute-phase proteins in cerebrospinal fluid (CSF) following SAH. Concentrations of interleukin (IL)- 1beta, IL-6, transforming growth factor-beta1 (TGF-beta1) and C-reactive protein (CRP) in CSF of 36 patients with SAH were measured by enzyme-linked immunoabsorbent assay (ELISA). Floating cells collected from the CSF were centrifuged four to six days after SAH, and examined immunohistochemically. Intracellular IL-1beta and IL-6 were examined by flow cytometric analysis. The molecular weight of TGF-beta1 in CSF of 30 patients was examined by Western blot analysis. The TGF-beta1 levels of patients who had undergone ventriculoperitoneal (VP) shunt (n = 19) was significantly higher than nonshunt group (n = 16). The CRP levels of VP shunt group was significantly higher than nonshunt group. IL-6 concentration was maximal within day 0-1 and it was secreted by neutrophils and monocytes. ELISA showed consistently low levels of IL-1beta, whereas a proportion of monocytes and lymphcytes were IL- 1beta-positive by flow cytometric analysis. TGF-beta1 levels were also maximal on day 0-1 according to ELISA, although it tended to be in the inactive form derived from platelets. A 25 kDa band of TGF-1 was detectable for at least 13 days after SAH, which may have been secreted in part by neutrophils and monocytes. CRP levels in CSF peaked on day 2-3. The present results suggest that leukocytes induced by SAH play an important role in post-hemorrhagic inflammation in the subarachnoid space by releasing IL-6 and TGF-beta1. The CRP and TGF-beta1 levels in CSF are strongly concerned with communicating hydrocephalus after SAH.

Adult↗

[A case of IgG4-lambda type monoclonal immunoglobulin that interfered with determinations for albumin, direct bilirubin and iron in serum].

We report a human IgG4-lambda type M-protein that reacts with reagents of albumin, direct bilirubin and iron. Blood chemistry findings by an automated analyzer were: total protein 8.8 g/dl; albumin(ALB) 9.2 g/dl; direct bilirubin(D-Bil) -6.2 mg/dl; and iron(Fe) 25 micrograms/dl. The white precipitates were formed by the interaction of the patient's serum with reagents of ALB, D-Bil, and Fe. Immunofixation electrophoresis revealed that these precipitates were the IgG4-lambda type M-protein. Western blotting analysis showed that the IgG4 molecules were composed of two 55 kDa gamma 4 chains and two 24 kDa lambda chains.

Aged↗

New guiding catheter for transrad PTCA.

A new guiding catheter for PTCA is described. In our department, 302 patients (405 lesions) underwent transradial coronary angioplasty using the 6 Fr Kimny guiding catheter since January 1996. The total engagement rate using the Kimny guiding catheter was 91.3% (370/405). The engagement rate after the modified Kimny guiding catheter was introduced in May 1996 increased to 96.0% (243/253). The stent delivery success rate was 98.4%. We had two dislodged stents. PTCA for both left and right coronary arteries in a single procedure with the Kimny guiding catheter was performed via the radial artery in 27 patients. In 24 of these patients (89%) we engaged both coronaries successfully. In the remaining 3 patients we switched to another catheter. Except for 4 patients with non-Q-wave myocardial infarction, no major cardiac complications were encountered. No major entry site-related complications were seen, and no patient required vascular surgery or blood transfusions. In one patient the Kimny guiding catheter tip caused a minor dissection of the LMT, but no ischemic event occurred as a result. In conclusion, the Kimny device is a useful PTCA guiding catheter for routine angioplasty and stenting.

Aged↗

[Studies on the blood coagulation factor XIII in patients with increased levels of FDP D-dimer].

Antigen levels of blood coagulation factor XIII (XIII) were determined in plasmas from patients with increased levels of fibrin degradation products-D-dimer (FDP-DD), including disseminated intravascular coagulation (DIC), by latex photometric immunoassay using polyclonal anti-XIII a subunit antibody-coated latex reagent. Since stable fibrin is directly degradated by plasmin and FDP-DD is produced, plasma FDP-DD levels correlate with plasmin-alpha 2-plasmin inhibitor complex levels, but not with thrombin-antithrombin III complex (TAT) or XIII levels. In order to clarify other causes of discordant relationships among the related three parameters, we studied the changes in plasma XIII, TAT and FDP-DD levels in fourteen DIC patients induced by various primary disorders. Only in two cases, XIII levels changed up and down irrelevant to the fluctuating levels of TAT and FDP-DD. In seven cases, plasma XIII levels remained low during the clinical courses, indicating possibilities that elevated condition of XIII consumption continued and/or production of XIII was low. On the other hand, in four patients, including two patients with nephrosis, XIII might be produced at higher rate than that of consumption. Same phenomenon was observed in one of eight recipients with living-related liver transplantation who showed remarkably increased levels of FDP-DD without DIC. In conclusion, plasma XIII level in patients with elevated FDP-DD may be influenced by the balance between consumption of XIII by unstable fibrin and/or surgical stress and the following tissue recovery etc. and production of XIII in liver, megakaryocytes and monocytes.

Disseminated Intravascular Coagulation↗

[Detection of anti-granulocyte antibodies using flow cytometry].

To detect anti-granulocyte antibodies (AGAs) in the sera of granulocytopenic patients is important to study the mechanism of the disease. In this report, we studied neutrophil associated immunoglobulin (NAIg) and neutrophil binding immunoglobulin (NBIg) in patients' sera using flow cytometry (FCM). We investigated the interference of circulating immune complexes (CIC) on measuring the NAIg and NBIg. No apparent effect of CIC was observed at concentrations up to 140 micrograms/ml. NAIg and NBIg were semiquantitated by determining the relative fluorescence intensity (RFI) on a flow cytometer and the normal ranges of NAIg and NBIg were less than 15 RFI and less than 10 RFI, respectively. Of 100 sera from patients with neutropenia, 5 were positive for NBIg and 3 of them were positive for granulocyte-specific antibodies. One serum of a patient with benign chronic neutropenia showed clear specificity for NA1 alloantigen but the other 4 AGAs were not specific for NA alloantigen system. Our NAIg, NBIg screening procedure, including NA specificity testing of NBIg and detection of reactivity with normal lymphocytes using FCM, is simple and useful for measuring and studying serological and immunological characteristics of AGAs.

Autoantibodies↗

Distribution of retinol-binding protein in the human digestive tract.

By employing polyclonal antibodies for retinol-binding protein (RBP), its distribution in the human pancreas and digestive tract mucosa was compared with those of transthyretin (TTR) and various peptide hormones. The materials used included surgically removed pancreas, esophagus, stomach, small and large intestines. Paraffin sections were stained by the indirect immunoenzyme method. The results indicate that RBP-containing cells are found in the pancreas and the gastrointestinal mucosa, but most frequently in the gastric antrum and duodenum. In the pancreas, RBP-containing cells are found in the islets and among acinar and ductal epithelial cells, and consistently stain for chromogranin A. RBP-containing cells in the gastrointestinal mucosa showed typical features of endocrine cells and also stained for chromogranin A. The distribution of TTR in these tissue sites resembled that of RBP, but the immunoreactive intensities of both peptides altered independently. Comparison of the distribution of RBP, TTR, and various gastrointestinal peptide hormones revealed that the distribution of RBP coincided with none of the other peptides, although some of the RBP-containing cells stained for most of the peptides examined and vice versa. These results suggest that RBP may be a consistent component of gastrointestinal endocrine cells.

Antibodies↗

[The clinical usefulness of a new (the second generation) antibody to hepatitis C virus].

To clarify the clinical usefulness of a second generation antibody to hepatitis C virus (anti-HCV-2), we tested the anti-HCV-2 by enzyme immunosorbent assay (Imucheck HCV Ab) in comparison to the first generation antibody (anti-HCV-1). Serum samples were obtained from the patients with acute hepatitis, chronic liver diseases, alcoholic liver disease and autoimmune liver diseases. Furthermore, both antibodies were tested in serum samples from individuals in a hyperendemic area of non-A, non-B hepatitis. Anti-HCV-2 was detected earlier than anti-HCV-1 in acute type C hepatitis. The prevalence of anti-HCV-2 was higher than that of anti-HCV-1 in acute and chronic non-A, non-B liver diseases. However, prevalence of anti-HCV-2 was not increased in patients with non-viral liver disease such as pure alcoholic liver disease and autoimmune liver diseases. The HCV-infection rate was increased to about 40% by detection of anti-HCV-2 in individuals in hyperendemic area of non-A, non-B hepatitis. Thus, the assay of anti-HCV-2 is very useful to detect actual HCV infection. The accuracy and sensitivity of anti-HCV-2 were higher than those of anti-HCV-1.

Adult↗

[Anti-HCV and long-term prognosis in patients with non-A, non-B post-transfusion hepatitis].

Anti-HCV was tested in serial serum samples of 20 patients with non-A, non-B post-transfusion hepatitis (NANB-PTH) who had been followed for more than 3 years after the onset. The rate of chronicity of disease was significantly higher in anti-HCV positive patients (12 out of 14, 85.7%) than in anti-HCV negative patients (group 1) (1 out of 6 patients, 14.3%). Among 14 anti-HCV positive patients, 7 lost anti-HCV during the follow-up period (group 2) and 6 of these 7 patients revealed normalization of liver dysfunction before or after the disappearance of anti-HCV. To the contrary, remaining 7 patients whose anti-HCV sustained positive (group 3) did show persistent abnormal liver function tests. Anti-HCV titer was significantly higher in group 3 than in group 2 at the point of 1 year after PTH. The titration of anti-HCV at 1 year after the onset of PTH was thought to be useful for estimating the prognosis of patients with post-transfusion hepatitis type C.

Adult↗

[Analysis of cross-linked fibrin and fibrinogen degradation products with SDS-PAGE and immunoblot].

SDS-PAGE and immunoblot technique with anti-FDP-D, -FDP-E, -fibrinogen antibody or anti-FDP-DD monoclonal antibody were applied to analyze FDP fragments prepared from cross-linked fibrin and fibrinogen with plasmic digestion in vitro. FDP fragments of DY(260K), DD(187K), X(245K), Y(166K), D(77K, 97K), E1(58K), E2(46K) and E3(40K) were identified from data of molecular weight and reactivity to four antibodies referred to reports of other investigators. Serum FDP fragments from five DIC suspected patients were analyzed by the same methods. In two patients' sera, DD fragment was a main component, and in the other three patients' sera, D fragment was a main fraction. Proportions of high molecular weight fragments of FDP were considerably different in patients' sera. Appearance of D fragment in our cases was considered to be derived from unstable fibrin (fibrin monomer and dimer) rather than from fibrinogen. Molecular weight of DD fragments from patients' sera had heterogeneity (160 approximately 180K), and the values were different from that (187K) prepared from cross-linked fibrin. In conclusion, SDS-PAGE and immunoblot analysis of serum fragments of FDP will be an useful technique to investigate the clinical and pathological condition of DIC.

Disseminated Intravascular Coagulation↗

Seroepidemiology of hepatitis A, B, and D viruses and human T-lymphocyte tropic viruses in Japanese drug abusers.

To evaluate the prevalence of hepatitis virus markers and human T-cell lymphotropic virus infections among drug abusers in Japan, serum samples were collected from 91 male drug abusers at the Shinshu University Hospital and the rehabilitation facility in Matsumoto and from 519 healthy male blood donors as controls. Sera were tested for antibody to hepatitis A virus (anti-HAV), hepatitis B surface antigen (HBsAg), antibody to HBsAg (anti-HBs), antibody to hepatitis B core antigen (anti-HBc), immunoglobulin M anti-HBc (IgM anti-HBc), antibody to hepatitis D virus (anti-HDV), antibody to HTLV type 1 (anti-HTLV 1), and antibody to human immunodeficiency virus (anti-HIV). The prevalence of anti-HAV was 13.2% in drug abusers and 10.8% in controls (not significant). The prevalences of HBsAg, anti-HBs, anti-HBc and exposure rate to hepatitis B virus (HBV) were 4.4%, 24.2%, 31.9%, and 35.2%, respectively, in drug abusers and 0.8%, 6.7%, 9.6%, and 9.6% in controls. The exposure rate to HBV was significantly different (P less than 0.001). IgM anti-HBc and anti-HDV were not detected in any sera. Anti-HTLV I was detected in three drug abusers (3.3%) and in one (0.2%) of the controls (P less than 0.01). All sera were negative for anti-HIV in all subjects. Infection with HBV and HTLV I is more common among drug abusers than in the general population of blood donors in Japan.

Acquired Immunodeficiency Syndrome↗

[Purification of the Ro/SSA antigen and enzyme-linked immunosorbent assay for quantitation of the antibody].

Antibodies to the Ro/SSA antigen occur in patients with Sjögren's syndrome and other collagen diseases. In this study, we used ammonium sulfate precipitation, anti-Ro/SSA immunoaffinity chromatography and gel filtration to purify the Ro/SSA antigen from pig spleen. In addition, the normal human IgG affinity column chromatography was also used to adsorb nonspecific reacting materials as heterophile antigens which contaminated in preparations after immunoaffinity chromatography. On SDS-PAGE the purified Ro/SSA antigen migrated as 60K and 53K polypeptides bands and the antigenicity was observed on 60 K band. An enzyme-linked immunosorbent assay (ELISA) using the purified antigen was developed for quantitation of the antibody which was found to be sensitive, precise and specific. By the method, anti-Ro/SSA antibodies were detected in SLE (44%), MCTD (71%), PSS (50%), SjS (83%), RA (16%), PBC (23%), lupoid hepatitis (23%) but only 4% in normal controls. The purified antigen and ELISA method will provide beneficial tools for the investigation of autoimmune diseases.

Animals↗

[Triiodothyronine(T3) autoantibodies in a woman with nonthyroidal disorder: a study on preparation of serum IgG fraction employing protein A column chromatography].

A 48-year-old non-goitrous woman, who had undergone cardiac surgery for mitral stenosis under the extracorporeal circulation, showed high levels of serum T3 and free T3 in a recent follow-up study, employing antibody coated-bead RIA for T3 and -Amerlex M particle RIA for free T3. However, other thyroid function tests (T4, free T4, TSH and TBG) were normal. We suspected that thyroid hormone autoantibodies (THAA) in her serum interfered with T3 and free T3 analyses. The presence of THAA was demonstrated by the use of various procedures as follows. Firstly, the patient's serum was directly incubated with 125I-T3 or -T4 analog which did not bind to TBG, followed by B/F separation with polyethyleneglycol, counting the precipitates. Secondly, after the serum was treated with an acid-charcoal solution to remove circulating thyroid hormone, the measurement of THAA was made as stated above. Normal sera were used as controls. Both the non- and acid-charcoal-treated sera showed much higher percentages of 125I-T3 analog precipitation as compared with controls. In the case of 125I-T4 analog, there was no difference between them. In the third study, the presence of IgG antibodies that bound T3 but not T4 was investigated. The IgG fraction of the patient's serum was separated employing a Protein A-Sepharose CL-4B column chromatography. Then, the prepared IgG fraction was purified by a technique of gel filtration chromatography (Sephacryl S 200). Non-purified and purified-IgG fractions both revealed higher binding percentages of 125I-T3 analog than the control IgG fraction and non-IgG fraction of the patient. Furthermore, a good dose response was observed between the binding percentage of 125I-T3 analog and each dose of the patient's serum or IgG fraction. From these observations, it was clarified that this woman had anti-T3 IgG autoantibodies using a Protein A column chromatography with confirmation of gel filtration chromatography.

Autoantibodies↗

Presence of antideoxyribonucleic acid antibody in patients with hyperthyroidism of Graves' disease.

In 16 untreated patients with hyperthyroidism due to Graves' disease, serum antidouble stranded DNA antibody, measured by RIA, was positive (greater than 20 U/ml) in 14. In methimazole-treated patients with T3-suppressible thyroid uptake, anti-DNA antibody was found in 9% (3 of 35). The frequency of positive tests in methimazole-treated patients with T3-nonsuppressible thyroid uptake and in surgically treated patients was 24% (5 of 21) and 57% (4 of 7), respectively. Among anti-DNA antibody-negative (less than 9 U/ml) and weakly positive (10-19 U/ml) patients, those with T3-suppressible thyroid uptake had lower anti-DNA antibody titers than those with T3-nonsuppressible thyroid uptake. Among 32 patients with Hashimoto's thyroiditis, anti-DNA antibody was positive in 7. None of the patients with simple goiter had positive or weakly positive anti-DNA antibody results. Although the quantity of antibodies did not correlate well in individual patients, the rates of positive TSH binding-inhibiting immunoglobulin and anti-DNA antibody tests were roughly comparable in these patient groups. None of these patients with thyroid disease associated with anti-DNA antibody had clinical or other serological evidence suggestive of systemic lupus erythematosus or related collagen vascular disorders. The finding of anti-DNA antibody provides a new aspect of immunological abnormality associated with hyperthyroidism of Graves' disease.

Antibodies, Antinuclear↗