False negative results of anticardiolipin antibody test.
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Biomedical subjects
Publications and source records attributed to H Tsuda.
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Investigations on the caries-preventive effect of calcium fluoride have been seriously hampered by the lack of adequate detection techniques. In this paper, the micro-Raman technique is introduced as a suitable method for CaF2 quantification with a spot size typically 5 microns. Advantages of this measuring technique and associated problems are described for CaF2 determination in the presence of large amounts of hydroxyapatite. The data show that a comparative analysis of Raman intensities of calcium fluoride and phosphate ions of hydroxyapatite can be used for quantification of CaF2 concentrations. The intensity behaviour of the CaF2 and hydroxyapatite Raman lines is explained considering the absorption-scattering cross-sections of the two components. The ratio of the Raman intensity (322 cm-1) of CaF2 to the phosphate Raman intensity (432/447 cm-1) of hydroxyapatite was found to be related linearly to the ratio of the concentration of CaF2 to that of hydroxyapatite. At present, the lower limit of CaF2 quantification is approximately 3 wt%. The technique has already been applied on fluoridated bovine enamel, and Raman emissions due to CaF2 or CaF2-like material formed on/in enamel have been detected for the first time.
Raman spectra containing the distinct band at 322 cm-1 due to CaF2 or CaF2-like material formed in/on fluoridated bovine enamel were recorded using a micro-Raman spectrograph. Due to increasing levels of background fluorescence with increasing thickness of enamel, the Raman measurements were carried out on thin regions of wedged enamel sections. The distribution of the CaF2 or CaF2-like material was estimated using a simple model. The results indicate that 1/3 of the total CaF2 was concentrated within the narrow depth < 2 microns with high CaF2 concentrations (> 10 wt%), and that the majority of the CaF2 was distributed over the depths up to 26 microns (1 wt% CaF2). SEM observations on fractured fluoridated enamel confirmed that morphological changes were present in the depth range comparable to that of the high CaF2 concentration region expected from the Raman analysis. In deeper regions where lower concentration (< 10%) but a large amount of CaF2 was still expected, the SEM images failed to distinguish between the normal and fluoridated enamel. After KOH treatment, the Raman spectra did not show evidence of the CaF2 peak and the SEM micrographs also confirmed the removal of globules. The peak position of the Raman band of the CaF2 formed by the fluoridation was identical to that of pure CaF2. However, the linewidth was 23 cm-1 (FWHM) and a factor of 2 broader than that of pure CaF2 (12 cm-1). This implies that the lattice dynamics of the CaF2 formed by fluoridation is different from of pure CaF2, and that the material formed is 'CaF2-like' or 'disordered CaF2'.
Raman spectra of human dental calculus have been observed for the first time by use of micro-Raman spectroscopy. The spectral features of calculus were influenced easily by heating caused by laser irradiation. Therefore, the measurements were carried out at relatively low power (5 mW, 1-micron spot size). The spectra could be characterized as phosphate vibrational bands due to the v1, v2, v3, and v4 modes. The overall spectral features did not resemble those of pure minerals such as brushite, octacalcium phosphate, and hydroxyapatite. There were spectral differences among mixed calculus particles obtained from 18 adults, probably due to variations in local mineral composition and differences among patients. However, the averaged spectral features did not vary significantly with formation period from 1 to 6 months. Freshly removed and stored (5-11 months) calculus also gave comparable Raman spectra. Measurements on a fractured sample indicated that Raman spectra at saliva and dentin interfaces are nearly identical, and major mineral constituents may not vary significantly along the growth axis of calculus.
Retinal degeneration induced by a single parenteral dose (40 mg) of N-methyl-N-nitrosourea (MNU) was studied in rats and rabbits. Sequential observations showed that severe pathological changes, that is, complete or widespread destruction of the photoreceptor layer and the outer nuclear layer of the retina, occurred in both animals by 72 hr after the administration of MNU. At week 5, at the end stage of the retinal degeneration induced by MNU, thin remnants remained with the inner nuclear layer in direct contact with the pigment epithelium. An additional immunohistochemistry study using antibodies against 7-methyldeoxyguanosine (7-mdGua), revealed the presence of high levels of 7-mdGua DNA adducts in the nuclei of the outer nuclear layer cells. This finding suggests that the alkylation of the outer nuclear layer by MNU was the direct cause of the retinal degeneration seen in these animals.
Nagase analbuminemic rats (NARs) were compared to the Sprague-Dawley (SD) stock in a medium-term assay system for hepatocarcinogenesis regarding their susceptibilities to the influence of chemicals on the development of glutathione S-transferase, placental form, positive (GST-P+) foci. Two weeks after initiation with diethylnitrosamine (DEN), the animals were exposed alternatively to 0.06% 3'-methyl-4-dimethyl-aminoazobenzene (3'-Me-DAB), 50 ppm DEN, 0.25% ethionine, 1% clofibrate, and 1% butylated hydroxyanisole (BHA) for a 6-wk period. Adequate controls included groups only initiated with DEN or treated with each test compound alone. For evaluation of the modifying potential of the chemicals, indices were generated by using the mean values obtained for number and area of GST-P+ foci after each treatment. Comparison between these indices suggests that SD rats were relatively more sensitive than NARs to the modifying effects of complete carcinogens (3'-Me-DAB and DEN). The strains were similarly-susceptible to the promoting influence of ethionine, a nongenotoxic carcinogen. The inhibitory influence of BHA was more intense in NARs, whereas in both strains clofibrate was associated to similarly reduced values for number and area of GST-P+ foci. The degree of susceptibility of each strain to the modifying influence of chemicals on foci development depended on the chemical agent investigated.
During multistage liver carcinogenesis, there is a sequential decrease in gap junctional intercellular communication (GJIC), associated with reduced expression of a major liver gap-junction protein (connexin 32). There are also several lines of evidence indicating that the induction of cell proliferation plays an important role during liver carcinogenesis. The relationship between GJIC and cell proliferation and their roles in liver carcinogenesis are not yet known. Results from various experiments suggest that there is a close relationship between the inhibition of GJIC and stimulation of liver cell proliferation. However, our results also suggest that different stimuli may affect cell proliferation and GJIC differentially by different mechanisms.
Leukemia cells from ten patients with B-cell chronic lymphocytic leukemia (B-CLL) and one with B-CLL of mixed cell type (B-CLL/PL) were analyzed to delineate the cellular characteristics, especially the myelomonocytic nature. Most of the surface phenotypes were consistent with the previous reports. However, the frequency of positivity for CD22 and CD25 was much higher in the examined cases of B-CLL than in Western reports. B-CLL cells frequently possessed antigens which are mainly expressed in myelomonocytic cells: 8/10 for CD11b and 4/10 for CD11c, although the presence of CD14 was not apparent (0/10). In addition, colony stimulating factor-1 receptors (CSF-1R) were expressed in 6 of the 10 B-CLL cases. The reactivity with antigens such as CD14, CD11c, and CSF-R was markedly enhanced by in vitro incubation. Alterations in antigen expression and in in vitro survival by growth factors for myelomonocytic lineage were observed in certain cases. These data imply that not only are B-CLL cells phenotypically closely related to myelomonocytic lineage but that they may also be influenced by growth factors for that lineage.
We reported two cases with severe bleeding tendency. Coagulation tests revealed the development of high titer inhibitors to coagulation factor VIII. As results of immunological analyses, the inhibitor from case 1 was found to be IgG type autoantibody having both kappa and lambda light chains. The subclasses were IgG1 and IgG4. The inhibitor from case 1 recognized the COOH-terminal light chain (72kDa thrombin fragment) on the factor VIII molecule as an epitope. On the other hand, the inhibitor from case 2 recognized both epitopes of heavy chain (44kDa thrombin fragment) and light chain (72kDa thrombin fragment). The characteristics of the inhibitors demonstrated no difference between autoantibody and alloantibody from hemophilics as mentioned by Fulcher. These fragments (A2 and C2 domain) may be important on the function of factor VIII. On further progress of epitope analysis of inhibitors, it may be useful to know the structure-function relationship of factor VIII, and applicable to treat the inhibitor patients with some way such as extracorporeal adsorption using a peptide affinity column or peptide-induced neutralization.
Recently, many alterations in DNA were demonstrated in human breast cancer in vivo, and their clinical and pathological implications were extensively examined. Among these gene alterations, amplification of the c-erbB-2 proto-oncogene and mutation of the p53 tumor-suppressor gene, which occur relatively frequently and are accompanied by alterations in the respective protein levels, demonstrated strong correlation to the histologic grade of atypia and were shown or suggested to be prognostic factors independent of tumor size or lymph node status. Although further analyses are necessary to clarify their prognostic significance in patients without lymph node metastasis, these are suggested to be useful in predicting patient prognosis more accurately and aiding determination of adjuvant systemic therapy.
The disappearance of the response to single twitch stimulation (STS), double burst stimulation (DBS) and zero post-tetanic count (PTC) were evaluated to determine which best indicated the optimal time of endotracheal intubation during onset of neuromuscular blockade induced by vecuronium (0.08 mg kg-1) in 199 patients under thiopentone and halothane anaesthesia. Evaluations were performed by mechanomyographic and manual methods using a Myograph and a peripheral nerve stimulator. The study consisted of six parts. In part 1 (n = 30) and part 2 (n = 30), the response to STS (0.1 Hz) were evaluated mechanically and manually, respectively. In part 3 (n = 64), post-tetanic count (PTC) changes were evaluated mechanically. In part 4 (n = 30) and part 5 (n = 30), response to DBS were evaluated mechanically and manually, respectively. Intubation was performed immediately after obtaining zero PTC, or absence of response to STS (0.1 Hz) and DBS, and the intubation score was determined. In the control group of patients, intubation was performed under the same anaesthetic conditions as in parts 1-5 but without the administration of any muscle relaxant. The time from administration of any muscle relaxant. The time from administration of vecuronium until disappearance of response was 2.16 +/- 0.56, 2.49 +/- 0.68, 2.04 +/- 0.29, 2.20 +/- 0.53, and 2.97 +/- 0.61 min in parts 1-5, respectively. Excellent intubation conditions were not established in any of the control patients, and in 50.0, 70.0, 55.0, 70.0 and 90.0% of the patients in parts 1-5, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
A lot of alterations of oncogenes have been reported to occur in human cancers in vivo. The types of alterations which occur most frequently are point mutations of ras genes, amplifications of myc and erbB gene families and rearrangement of oncogenes by chromosomal translocation. Each type of gene alteration discloses the specificity not only to organ and cell types from which cancer originated but also to etiological background and clinical aggressiveness of cancer. These gene alterations are shown to activate oncogenes and suggested to be involved in the mechanism of genesis and/or progression of human cancers.
Deletion mapping of chromosome 3p was performed on 47 cases of human uterine cervical cancer using 24 polymorphic DNA markers including five inter-Alu DNA markers and two NotI-boundary cosmid markers obtained in our laboratory. The most likely order of these 24 polymorphic DNA markers was determined as being cen-[D3S4, H8]-D3S693-D3S659-D3S30-D3S687-[D3S2, UR9, UR47]-J36-J17-GNAI2B-D3F15S2-D3S643- D3S32-D3S23-D3S686-H35-UR189-D3S685-D3S 11 - D3S12-THRB-D3S22-pter, based on the data from radiation hybrid mapping genetic linkage analysis and in situ hybridization. Loss of heterozygosity (LOH) at one or more loci on chromosome 3p was detected in 21 of 47 cases (45%). Four tumors showed partial or interstitial deletions, and the common region of LOH in these tumors was 3p13-p21.1 between the D3S30 marker and the D3S2 marker. Candidates for tumor-suppressor genes, APEH, D8, GNA12B, ZNF35, RARB, THRB and RAFI, were all mapped outside of the common region in uterine cervical cancer. However, this region is commonly deleted in carcinoma of the lung, breast and kidney, and encompasses the breakpoint of the (3;8) translocation in hereditary renal cell carcinoma. This result indicates the presence of a novel tumor-suppressor gene in the region of 3p13-p21.1, which is involved in the development of several human cancers.
A case of an 18-year-old female, who developed an intraventricular hemorrhage and recurrent cerebral ischemic r attacks with Moyamoya like angiographical findings was reported. Lupus Anticoagulant (LA) was present in the patient's plasma. Cerebral angiography demonstrated stenosis and occlusion of the intracranial major arteries. Hemorrhagic episodes in patient's with LA is quite rare. The mechanism of intraventricular hemorrhage and the significance of LA in the differential diagnosis of Moyamoya disease are discussed.
We tested whether parvovirus B19 (PVB19) is involved in benign lymph node swelling as a causative agent. The serum specimens from 13 patients with lymph node swelling of unidentified cause were tested for IgM and IgG antibody against PVB19 by Western blot analysis. Five patients had IgM antibody, and PVB19 DNA was detected in the serum from one of them by dot-blot analysis, suggesting the recent PVB19 infection and its involvement in lymph node swelling of these patients. Clinical and haematological features of the patients are also discussed.
When B-cell chronic lymphocytic leukemia (B-CLL) cells derived from peripheral blood are cultured in vitro, a substantial proportion die spontaneously by programmed cell death (PCD). In this study, we reexamined this phenomenon with both B-CLL and T-CLL cells, using a serum-free culture system. After 48 h of culture, in B-CLL cells, 23.3% +/- 6.7% (mean +/- S.E.M., n = 3) of total DNA was fragmented, while in 2 cases of T-CLL, this value was 32% and 45%, respectively. Analysis by electrophoresis indicated that the DNA of the B-CLL and T-CLL cells had been cleaved into regular oligonucleosome fragments comprising approximately 180-200 base pairs. This process was significantly promoted by methylprednisolone and the protein kinase A (PKA) activator Sp-cAMPS in at least some cases. Since B-CLL and T-CLL cells often possess interleukin-2 (IL-2) receptors on their cell membranes, we assessed the effects of IL-2 on spontaneous PCD. After a 48-h culture PCD was inhibited by 100 units/ml IL-2 by 31.7% +/- 6.6% in B-CLL (n = 3) and by 47% in one of 2 cases of T-CLL. This protective effect of IL-2 against spontaneous PCD was dose- and time-dependent. These findings suggest that the viability of B-CLL and T-CLL cells in vivo is regulated both positively and negatively by intrinsic IL-2, glucocorticoids, and regulators of PKA activity. This process of cell death may be involved in the development of CLL.
The following developments have considerably influenced the pathological management of breast materials: The wide use of aspiration cytology, increase of specimens from small non palpable lesions detected by mammography, an increase of cases difficult to diagnose histologically, increase in the number of conservative surgery cases which require examination of semi-serial sections, and the development of immunohistochemical procedures using breast cancer tissues. Under these changing circumstances in the department of pathology, we report our current practices and some of the results; About 75% of breast cancer cases initially treated in our hospital were diagnosed by aspiration cytology only. A retrospective, simulation study on quadrantectomy of breast cancers for selected T1N0N1a examined by semi-serial sections in non-conservative mastectomy cases revealed residual carcinoma foci located in a region outside the primary quadrant in about 25% cases. GCDFP-15 (Gross cystic disease fluid protein-15) is a fairly specific marker for breast cancer. Good correlation was obtained using estrogen and progesterone receptors routinely measured by ICA (Immunocytochemical assay), instead of the biochemical method using DCC (Dextran-coated charcoal). c-erbB-2 and p53 which were positive in 45.8%, and 28.6%, respectively, in our breast cancer cases, seemed to be good markers of biological malignancy in the breast cancer.
Currently, many tumor markers are routinely measured in patient serum, but none appears to adequately detect cancer-specific substances for breast cancer. Four tumor markers (CEA, CA15-3, BCA225, c-erbB-2) were compared for sensitivity and specificity to breast cancer in 393 patients with breast disease (fibrocystic disease; 40, benign tumor; 21, primary cancer; 22, recurrent or advanced cancer; 22, non-recurrent; 288), and the following results were obtained: In cases of primary breast cancer excluding stage IV cases, the positive rates of CEA, CA15-3 and BCA225 were 4.5%, 13.6% and 13.6%, respectively. These rates were not higher than the positive rates found in fibrocystic disease or benign tumor. In cases of recurrent or advanced breast cancer, the positive rates of CEA, CA15-3, BCA225 and c-erbB-2 were 31.8%, 50.0%, 40.9% and 27.3%, respectively. In the recurrent cases, the combination assay using CA15-3, BCA225 and c-erbB-2, showed more useful diagnostic value (72.7%) than other combination assays with three tumor markers. High levels of CA15-3 in eight cases, BCA225 in five and CEA in one were found during the follow up of 15 patients with initial recurrence. Serum c-erbB-2 exceeded the normal range in 6 of 7 cases with advanced breast cancer. Serum c-erbB-2 should be considered a marker of progressive breast cancer. These results suggested that CA15-3, BCA225, c-erbB-2 and combination assays are useful tumor markers for not only detecting the recurrence of breast cancer, but also diagnosing the progression of primary breast cancer.