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Biomedical subjects

M Kamachi

Publications and source records attributed to M Kamachi.

At least 37 records · Page 2Linked to original sources
PubMed2000

Induction of COX-2 expression by nitric oxide in rheumatoid synovial cells.

Prostaglandins formed by cyclooxygenase (COX) enzymes are important mediators of inflammation. The contribution of inducible COX-2 in the rheumatoid synovium is well documented. In this study, we evaluated the contribution of nitric oxide (NO) to COX-2 expression in rheumatoid synovial cells. Exposure of rheumatoid synovial cells to a NO donor, SNAP, induced COX-2 protein expression in a dose-dependent manner. RT-PCR analysis also demonstrated that COX-2 mRNA was induced in SNAP-treated synovial cells. Dexamethasone at therapeutic concentrations markedly inhibited this NO-mediated COX-2 expression in synovial cells. In contrast to its effect on COX-2 expression, SNAP did not affect the constitutive expression of COX-1 in rheumatoid synovial cells. Our findings suggest that NO is an important modulator of COX-2 expression and that glucocorticoids exert their anti-inflammatory action in rheumatoid synovium, at least in part, by suppression of COX-2 induction.

Arthritis, Rheumatoid↗
PubMed2000

Expression and function of X chromosome-linked inhibitor of apoptosis protein in Sjögren's syndrome.

Apoptotic cell death in acinar and ductal epithelial cells is thought to play an important role in the development of salivary gland dysfunction in patients with Sjogren's syndrome (SS). We examined the expression of anti-apoptotic molecules in salivary glands from patients with SS. The labial salivary glands from six human T-cell leukemia virus (HTLV)-I-seronegative and eleven HTLV-I-seropositive SS patients were analyzed by immunohistochemistry. In vitro experiments were performed with a human salivary gland cell line (HSG cells). Immunohistologic analyses revealed that Bcl-2 and Bcl-x were preferentially expressed in salivary infiltrating mononuclear cells more than acinar and ductal epithelial cells. In contrast, strong X chromosome-linked inhibitor of apoptosis protein (XIAP) expression was evident in both acinar and ductal epithelial cells. The pattern of expression of these anti-apoptotic molecules was similar in both HTLV-I-seropositive and HTLV-I -seronegative SS patients. Western blot analysis confirmed expression of XIAP in cultured HSG cells. The expression of XIAP in HSG cells was increased by IL-1beta, TGF-beta1, or IL-10. However, XIAP expression was down-regulated by TNF-alpha, which induced apoptotic cell death of HSG cells with an increase in caspase-3 activity. These effects of TNF-alpha in HSG cells were antagonized by IL-1beta, TGF-beta1, or IL-10. Our results suggest that XIAP is important in regulating apoptotic cell death of acinar and ductal epithelial cells in patients with SS.

Adult↗
PubMed2000

[A case of non-small-cell lung cancer with intramedullary spinal cord metastasis diagnosed pre-mortem].

The patient was a 54-year-old man who in May 1999 received a diagnosis of squamous cell carcinoma, T4 N2 M1, stage IV. Systemic chemotherapy and stereotactic radiosurgery were performed only to result in further progression of the disease. In August 1999, he experienced gait disturbance due to lumbar pain. Rehabilitation improved the gait disturbance and he was discharged. In October, since the pain reappeared and there was numbness in the right leg, he was readmitted. Brain MRI revealed multiple brain metastasis and whole brain irradiation was performed. But his symptoms deteriorated, and palsy of the right leg ensued. Later, bladder dysfunction also developed. Since spinal cord MRI revealed intramedullary metastasis at Th 12 and L1 levels, we performed radiotherapy for the lumbar medullary lesion, together with systemic chemotherapy. After chemoradiotherapy the tumor size decreased and the pain improved. Cases of lung cancer with intramedullary metastasis are rare, especially those diagnosed before death.

Brain Neoplasms↗
PubMed1999

[A clinical application of laser scanning cytometer--the significance in the cytology as an additional diagnostic technique].

Laser scanning cytometer (LSC) is a new machine similar to flow cytometer but with advantages for certain clinical and research applications. LSC is a microscope based and measures cells on a slide with the position of each cell on the slide. This new technique of LSC can be utilized on extremely small specimens and enables direct correlation of all of the measured fluorescent parameters with light microscopic cytologic morphology. To date, LSC has been successfully used to perform DNA content analysis of numerous specimen types and automated analysis of fluorescence in situ hybridization specimens. In this report, we describe characteristics of LSC comparison with flow cytometry and a clinical application of LSC focused on DNA content analysis in clinical specimens with pulmonary disorders. LSC provides a number of benefits that may make it more suitable for clinical laboratories than FCM.

Bronchi↗
PubMed1999

Control of photoinduced electron transfer from zinc-porphyrin to methyl viologen by supramolecular formation between monoclonal antibody and zinc-porphyrin.

Photoinduced electron transfer from tetrakis(4-carboxy-phenyl porphyrin)-zinc complex (Zn-TCPP) to an acceptor molecule (methyl viologen; MV2+) has been found to be controlled by the complex formation of monoclonal antibody 03-1 for the porphyrin (TCPP) and Zn-TCPP. Although there are no ground-state interactions between Zn-TCPP and MV2+ for a 2:1 complex of antibody 03-1 and Zn-TCPP, the fluorescence of Zn-TCPP is quenched by the addition of MV2+. The Stern-Volmer plots and emission lifetime studies show that there is a long-range electron transfer through the antibody 03-1.

Antibodies, Monoclonal↗
PubMed1998

[Delays in recognizing facial expressions resulting from prolonged viewing of adaptation face stimuli].

The main purpose of the present study was to investigate the mechanism underlying fundamental dimensions in recognition of facial expressions, using the prolonged viewing method. Pictures of female faces were used, which displayed one of the following six: happiness, sadness, surprise, anger, disgust, and neutral expressions. In Experiment 1, the mean ratings of each expression on each picture were analyzed with principal component analysis (PCA). In Experiment 2, subjects orally judged the expressions of test faces following either one or 25 seconds of viewing of an adaptation face. Significant delays occurred in the prolonged viewing condition only when the adaptation face had a higher absolute value on 'pleasantness' the first component of PCA, than the test face. In contrast, adaptation faces that were high only on 'arousal,' the second component, produced no such effects. It was suggested that there are at least two subsystems involved in the recognition of facial expressions and that each system has different temporal characteristics.

Adaptation, Psychological↗
PubMed1997

Structural characterization of mouse monoclonal antibody 13-1 against a porphyrin derivative: identification of a disulfide bond in CDR-H3 of Mab 13-1.

The amino acid sequence of a mouse monoclonal antibody Mab13-1, a catalytic antibody against TCPP (meso-tetrakis(4-carboxyphenyl)porphyrin), was confirmed by mass spectrometric (MS) peptide mapping. The amino-terminal sequence of the heavy chain was established by MS/MS analysis of the isolated N-terminal peptide. The presence of a unique disulfide bond between Cys93H and Cys102H was identified by MS peptide mapping and sequence analysis of an S-S containing peptide. Positions of other disulfide bonds were identified to be conserved. The non-conserved disulfide bridge was found to be resistant as other intra-chain disulfide bonds against reduction under non-denaturing condition, and to be buried inside the molecule. This extra disulfide bond is expected to support antigen-binding by restricting the flexibility of CDR-H3 loop, and it might be favorable for the recognition of a plane antigen, a porphyrin derivative.

Amino Acid Sequence↗
PubMed1996

Mucous gland adenoma of the trachea resected with an endoscopic neodymium: yttrium aluminum garnet laser.

We report a case of mucous gland adenoma of the trachea in a 73-year-old male revealed by bronchoscopy. The tumor was resected with a contact neodymium: yttrium aluminum garnet (Nd-YAG) laser after five years of observation. The tumor was histologically peculiar because it presented numerous cystically dilated, or irregularly shaped mucus-filled glands lined with cuboidal or tall columnar cells. In some parts, the lining cells of the tumor showed papillary proliferation. We diagnosed this tumor as a mucous gland adenoma of the trachea. We review the clinical features of this rare tumor and discuss the usefulness of the laser in the diagnosis and the therapy.

Aged↗
PubMed1996

[Clinical evaluation of hepatitis C virus RNA quantification by competitive reverse-transcription PCR].

We have quantified the hepatitis C virus (HCV) RNA by competitive reverse-transcription PCR method (Amplicor) HCV quantification monitor kit) in sera with positive HCV antibody measured by means of the second generation HCV antibody assay. Among the visitors to PL Tokyo Health Control Center for their health examination and the patients to National Defense Medical College Hospital, 123 HCV antibody-positive cases were examined. A positive but low correlation between the amount of HCV-RNA and the titer of HCV antibody (r = 0.508, p value < 0.0001) was obtained. HCV-RNA was not detectable in 19 HCV antibody-positive cases. Among them, 10 cases showed normal ALT values. In the cases with more than 1000 copies/ml of HCV-RNA, the greater the amount of HCV-RNA the higher ALT values were observed, while the titer of HCV antibody was not correlated to ALT. This study demonstrated the dissociation and low correlation between HCV-RNA amount and antibody-titers in some patients, which may recommend direct quantification of RNA for clinical evaluation of the patients with positive-HCV antibody. The quantification of HCV-RNA by such rapid and simple methods can be applicable for the determination of HCV-RNA amount in routine laboratory works.

Hepacivirus↗
PubMed1996

[Pathogenesis-screening tests for liver dysfunction in the asymptomatic patients with elevated ALT values and their diagnostic efficacies in primary care medicine].

We have evaluated the diagnostic efficacies of ultrasonography and hepatitis C virus (HCV) antibody measurement to differentiate pathogenesis of liver dysfunction in the asymptomatic adults with elevated ALT value. Among 4256 visitors to PL Tokyo Health Control Center for their health examination, 463 cases (11%) showed abnormal liver function including elevation of ALT value. Ultrasonography and HCV antibody measurement using the second generation reagent had been applied to 362 cases in order to screen the etiology of liver dysfunction. The ultrasonography succeeded to establish the diagnosis of fatty liver in 137 cases (38%) and 41 cases (11%) demonstrated positive HCV antibody. There were 4 cases with positive HBs antigen, however, it was found that their abnormal liver function was attributed to other etiology such as fatty liver and alcoholic liver dysfunction rather than chronic type B hepatitis. HCV antibody-positive cases showed higher levels of total protein, ZTT, AST, ALT, and lower levels of albumin, A/G, total cholesterol, triglyceride, gamma-GT and cholinesterase value than other cases. HCV antibody titers were not correlated to hepatic parenchymal damage estimated by ALT or cholinesterase value. Only a little correlation was observed between HCV antibody titers and HCV-RNA amounts determined by the competitive reverse transcription-polymerase chain reaction (RT-PCR) method. These results indicate sufficient diagnostic efficacies of ultrasonography and HCV antibody measurement for a pathogenesis differentiation in the asymptomatic patients with liver dysfunction, and these examinations should be employed as the first-step screening tests for the etiology determination of liver diseases in the primary care medicine.

Alanine Transaminase↗
PubMed1995

Thermostable peroxidase activity with a recombinant antibody L chain-porphyrin Fe(III) complex.

In order to engineer a new type of catalytic antibody, we attempt to use a monoclonal antibody L chain as a host protein for a porphyrin. TCPP (meso-tetrakis(4-carboxyphenyl)porphyine) was chemically synthesized and Balb/c mice were immunized using TCPP as a hapten. Two hybridoma cells (03-1, 13-1), that produce monoclonal antibody against TCPP, were obtained. Genes for both H and L chains of monoclonal antibodies were cloned, sequenced and overexpressed using E. coli as a host. ELISA and fluorescence quenching method show that the independent antibody L chains from both Mab03-1 and Mab13-1 have specific interaction with TCPP. Furthermore, the recombinant antibody L chain from Mab13-1 exhibits much higher peroxidase activity than TCPP Fe(III) alone. The enzyme activity was detectable with pyrogallol and ABTS (2,2-azinobis-3-ethylbenzthiazolin-6-sulfonic acid) but not with catechol. This new catalytic antibody was extremely thermostable. Optimum temperature of the peroxidase reaction by the complex of 13-1L chain and TCPP Fe(III) was 90 degrees C, while that the TCPP Fe(III) alone was 60 degrees C.

Amino Acid Sequence↗
PubMed1994

[Analysis of nuclear DNA heterogeneity of the hepatocellular carcinoma (HCC)].

To evaluate the objective proliferative activity in HCC nuclear DNA contents were measured by means of microspectrophotometry and at the same time the immunohistochemical technique using anti-PCNA antibody was employed. Surgically resected 26 HCCs were analyzed in terms of cell proliferative activity and regional heterogeneity. The analysis was performed by immunohistochemical demonstration of PCNA and pathologic histochemical study in formalin-fixed, paraffin-embedded specimens and cytophotometric measurements of nuclear DNA contents in fresh specimens. The results were as follows. 1) Nine HCCs showed regional ploidy heterogeneity. 2) PCNA labeling index and histological grade of the marginal area was much higher than that of the central area. From these results, we concluded that in the process of the HCC progression proliferative activity was decreased in the central area and was not decreased in the marginal area.

Carcinoma, Hepatocellular↗
PubMed1994

[Microspectrophotometric analysis of nuclear DNA contents related to the presence of alpha-fetoprotein in human hepatocellular carcinoma].

A total of 28 surgically resected hepatocellular carcinomas, including both fresh-smeared specimens and paraffin-embedded tissues, was stained with alpha-fetoprotein (AFP) antibody. In all cases we measured the DNA contents using microspectrophotometry. In 5 cases, analyses of the nuclear DNA histogram of both the AFP-positive and negative cells from the same specimen were successfully performed. The ratio of the S-G2.M phase of the AFP-positive cells was 2.24 times more than that of the AFP-negative cells. In addition, not only the ratio of S-G2.M phase cells, but also the incidence of aneuploidy pattern of AFP positive stained cases was higher than the negative stained cases. Histologically, the AFP-positive cases contained various differentiated tissues more than the negative cases. From these results, we confirmed that AFP production was related to the growth of HCC, and the results suggest that AFP producing HCC have a higher potential for growth than AFP non-producing HCC.

Carcinoma, Hepatocellular↗
PubMed1991

Facile assignment of sequence ions of a peptide labelled with 18O at the carboxyl terminus.

The combination of collision-induced dissociation (CID) and linked-scan analysis was used for analysing the sequence ions from the precursor ion of a peptide, which had been labelled with 18O at its carboxyl terminus (C-terminus) using 40 atom % H2 18O. The CID and linked-scan mass spectrum of the labelled peptide gave two series of sequence-ion signals: the one, originating from the C-terminus of the labelled peptide, showed a doublet signal due to the part-incorporation of 18O into the carboxyl group at the C-terminus, while the other, originating from the amino terminus (N-terminus), has the natural isotopic ion distribution. From the distribution of the isotopic ions in a single CID spectrum, the sequence ions containing the C-terminus could be readily differentiated from those containing the N-terminus, allowing the facile assignment of sequence ions to the amino-acid sequence of a peptide by CID and linked-scan analysis. This method was successfully applied to determination of the amino-acid sequence of the light-chain of mouse anti-porphyrin monoclonal antibody.

Amino Acid Sequence↗
PubMed1990

Computed tomography and nonoperative treatment for blunt abdominal trauma.

Studies were undertaken to determine if computed tomography (CT) could reliably assist physical examination in the initial assessment of blunt abdominal trauma, and also to examine how various abdominal injuries were managed with the guidance of CT. A total of 255 patients underwent emergency abdominal CT following blunt abdominal trauma over a period of seven years. One hundred and fifty two patients had abnormal CT scans, including 58 hepatic, 36 renal, 25 splenic and 9 pancreatic injuries as well as 67 patients with intra-abdominal hemorrhage and 21 patients with free abdominal air. A comparative study on the detection of pneumoperitoneum revealed CT to be far superior to plain radiography. One hundred and three patients had normal CT scans, all of whom were managed nonoperatively, except for three false-negative cases and two nontherapeutic cases. The patients with injury to the parenchymal organs were given nonoperative treatment if they had stable vital signs and no evidence of associated injuries demanding immediate surgery and the majority of these patients were managed well nonoperatively. CT was thus found to be a useful adjunct in the management of victims of blunt abdominal trauma, since in a rapid and noninvasive fashion, CT accurately defined the extent of parenchymal organ injury and also disclosed any other abdominal injuries.

Abdominal Injuries↗
PubMed1988

Immunocytochemical presentation of alpha-fetoprotein-producing gastric cancer in ascitic fluid: a case study.

A case of primary gastric cancer without hepatic metastasis showing extremely high alpha-fetoprotein (AFP) levels is reported. This case illustrates the application of the immuno-peroxidase technique to ascitic fluid cytology. Papanicolaou-stained smears of the ascites permitted the diagnosis of a metastatic carcinoma. A positive reaction to AFP was demonstrated in the tumor cells in the ascitic fluid cellular samples as well as in the paraffin-embedded tissue section of the primary gastric carcinoma. Rising AFP levels were also detected in ascitic fluid. AFP fractionation using lectin-affinity-crossed-line immunoelectrophoresis showed the hepatic rather than yolk sac type. Reports of such occurrences are few; no study, to the best of our knowledge, has previously documented cytological and immunocytochemical diagnosis in ascitic fluid. AFP-producing gastric cancer should be considered in the differential diagnosis.

Adenocarcinoma↗