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Biomedical subjects

K Uchida

Publications and source records attributed to K Uchida.

At least 181 records · Page 10Linked to original sources
PubMed2001

[Development for the determination of band 5b tartrate-resistant acid phosphatase activity in serum].

Band 5 Tartrate-resistant acid phosphatase(TRACP; EC 3.1.3.2) consists of two isoenzymes, bands 5a and 5b, of which band 5b TRACP, an enzyme expressed in bone-resorbing osteoclasts, is secreted into the circulation during bone resorption. Band 5b TRACP was measured kinetically in serum as tartrate-resistant fluoride-sensitive heparin-resistant ACP with 2,6-dichloro-4-acetylphenyl phosphate as substrate at pH6.6. The within-run(n = 20) and between-run(n = 20) CVs of band 5b TRACP activity were 3.3-5.8% and 5.0-7.3%, respectively. The reference range of band 5b TRACP activity in males(n = 72) and females(n = 87) 20-39 years of age by this method were 3.7-12.5 U/l and 2.7-9.9 U/l, respectively. The band 5b TRACP value was significantly higher in post-menopausal women compared with the menstruating women. The relationship of band 5b TRACP and ultrasound findings in healthy women aged 31-75 years(n = 139) were inversely correlated with stiffness(r = -0.401), speed of sound(SOS; r = -0.386) and broadband ultrasound attenuation(BUA; r = -0.338). These results suggest that band 5b TRACP may be a useful in the evaluation of bone turnover.

Acid Phosphatase↗
PubMed2001

Production of recombinant human creatine kinase (r-hCK) isozymes by tandem repeat expression of M and B genes and characterization of r-hCK-MB.

BACKGROUND: Serum creatine kinase-MB isoenzyme (CK-MB) is widely used as a marker of myocardial injury. We prepared recombinant human CK (r-hCK) MB isoenzyme and examined its potential for use as a control material for assay of CK-MB in serum. METHODS: cDNAs encoding CK-M and CK-B subunits were inserted into the same plasmid vector, followed by transformation of Escherichia coli. The resulting three types of CK isoenzymes were purified by conventional chromatography. RESULTS: The ratio of MB to MM to BB was 50:40:10 on the basis of CK activity. Highly purified CK-MB with a specific activity of 533 U/mg was produced in a yield of 5.7 mg/g of packed cells. Purified r-hCK-MB had the isoelectric point (pI 5.3) and molecular size (46 kDa for the subunit) of native CK-MB. Its immunoreactivity in an ELISA using antibody against native heart enzyme was similar to that of cardiac CK-MB. The r-hCK-MB retained >90% activity for at least 4 months at 11 degrees C in a delipidated serum matrix in a liquid form at a concentration of 118 U/L. CONCLUSIONS: r-hCK-MB shows key properties of the native cardiac isoenzyme and may be useful as a control and calibrator for serum assays of CK-MB.

Creatine Kinase↗
PubMed2001

[Sentinel node detection of patients with breast cancer by radionuclide method: consideration of radiation safety].

Sentinel node was detected by 99mTc labeled nanocolloid in five patients with breast cancer. Surgery of breast cancer was done at 16 hours after the administration of 74 MBq of 99mTc labeled nanocolloid. Sentinel node was searched by scintigraphy prior to surgery and by gamma-probe during surgery. Radioactivity of injected site, sentinel nodes, blood contaminated gauze, and other garbage was measured by GM detector. Radiation to medical staffs was monitored by a pocket radiation detector and film batches. Sentinel nodes were successfully detected both by scintigraphy and gamma-detector. More than 70% of radioactivity remained in the administered site at 16 hours. Small amount of radioactivity was detectable in the sentinel node. Almost no radioactivity was detectable in blood-contaminated gauze and other garbage. Radiation dose to the main surgeon was 4 to 6 microSv per surgery by a pocket radiation detector. Radiation dose to the assistant surgeon was 2 microSv per surgery. Radiation dose by labeling or injection was 0 to 1 microSv per procedure. No detectable radiation was measured by film batches. It is concluded that the detection of sentinel node by 99mTc labeled nanocolloid is a safe procedure from the point of radiation safety consideration.

Breast Neoplasms↗
PubMed2001

Argpyrimidine, a blue fluorophore in human lens proteins: high levels in brunescent cataractous lenses.

PURPOSE: To determine whether the human lens contains argpyrimidine, a modification of arginine by methylglyoxal, to establish how argpyrimidine content relates to lens aging and cataract formation. METHODS: A monoclonal antibody was used to measure argpyrimidine by a competitive ELISA in water soluble (WS) and insoluble (WI) lens fractions from young, aged, nuclear cataractous, and brunescent cataractous lenses. Brunescent cataractous lens proteins were digested by enzymes, the digest was subjected to HPLC, and the eluate was analyzed for argpyrimidine. Lens proteins from aged lenses (from donors 65 to 80 years of age) were fractionated on a Sephadex G-200 column, and the crystallins were tested for argpyrimidine. RESULTS: The competitive ELISA showed two to three times as much argpyrimidine in water-insoluble proteins as in water-soluble proteins. Although no clear cut increase with the age of the lens donors in either the water-soluble or the insoluble protein fractions was found, the argpyrimidine levels in brunescent cataractous lenses were significantly higher (254.0 +/- 155 pmol/mg protein, P < 0.005) than in age-matched, aged (16.1 +/- 8 pmol/mg) or nuclear cataractous lenses (49.0 +/- 26 pmol/mg). Lenses from diabetic individuals showed a modest increase (50.3 pmol/mg) compared with age-matched normal lenses. HPLC results provided additional evidence that human lenses contain argpyrimidine. Western blotting experiments showed consistently stronger reactions with cataractous lens proteins than those from noncataractous lenses, and argpyrimidine was found in both crystallin monomers and polymers. All crystallins and several cross-linked high-molecular-weight aggregates reacted with the antibody to argpyrimidine, but a protein of approximately 28 kDa in the alpha-crystallin fraction displayed the greatest immunoreactivity. CONCLUSIONS: Methylglyoxal modifies arginine within the human lens, and the changes occur at a much higher rate in brunescent lens proteins than in either nuclear cataractous or normal lenses. All crystallins contained argpyrimidine and covalently cross-linked aggregates. This is the first report of immunologic evidence for an arginine modification in the human lens by a physiologically important alpha-dicarbonyl compound.

Adolescent↗
PubMed2001

Evaluation of prognostic factors for myeloperoxidase anti-neutrophil cytoplasmic antibody- (MPO-ANCA) associated glomerulonephritis.

AIMS: To identify prognostic factors for myeloperoxidase anti-neutrophil cytoplasmic antibody- (MPO-ANCA) associated glomerulonephritis. MATERIALS: We analyzed the relations between the clinical and histological features of MPO-ANCA-associated glomerulonephritis and clinical outcome in 14 patients with the disease. The patients were divided into two groups: group 1 consisted of 5 patients with progressive deterioration of renal function leading to end-stage renal disease or chronic dialysis, group 2 consisted of 9 patients in whom the initial deterioration of renal function had improved by the time of the final examination. RESULTS: Creatinine clearance at the time of biopsy was significantly lower in group 1 than in group 2, and urinary protein was higher. The mean interval between onset of symptoms and biopsy in both groups was almost the same. Recovery of renal function was correlated with the percentage of global sclerosis, but patients who had severe crescent formation did not always have a poor response to steroid therapy. There was no statistical difference between the two groups in treatment regimens. Four patients required hemodialysis at the time of biopsy (3 in group 1 and 1 in group 2). Plasmapheresis was performed in 5 patients (1 in group 1 and 4 in group 2). CONCLUSIONS: Degree of proteinuria and renal dysfunction are indicators of a poor prognosis in MPO-ANCA-associated glomerulonephritis. Global sclerosis is a histological feature that is an indicator of a poor prognosis, whereas cellular crescent formation is a predictor of a good response to steroid therapy.

Adult↗
PubMed2001

An oral drug delivery system targeting immune-regulating cells ameliorates mucosal injury in trinitrobenzene sulfonic acid-induced colitis.

Control of immune-regulating cells in the colonic mucosa is important in the treatment of patients with inflammatory bowel disease (IBD). The aim of study was to examine the therapeutic effect of dexamethasone (DX) microspheres on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in rats, a model for human Crohn's disease. DX microspheres and DX alone were administered orally to rats with TNBS-induced colitis. The macroscopic score, histological score, myeloperoxidase (MPO) activity, nitric oxide (NO) production, and gene expressions of proinflammatory cytokines, cyclooxygenase (COX)-1, and COX-2 in the colonic tissue were determined. Proliferating cell nuclear antigen (PCNA) staining and expression of nuclear transcription factor (NF)-kappaB in colonic tissues were also investigated. Macroscopic score, histological score, MPO activity, and NO production in rats treated with DX microspheres were significantly lower than in those treated with DX alone. The gene expression of proinflammatory cytokines and COX-2 in rats treated with DX microspheres was down-regulated, compared with that in rats treated with DX alone. The number of PCNA-positive cells in the DX microsphere group was larger than in the group treated with DX alone. DX microspheres suppressed NF-kappaB activation in TNBS-induced colitis more strongly than DX alone. Oral administration of DX microspheres appears to ameliorate mucosal injury in TNBS-induced colitis. This drug delivery system could be an ideal therapy for human IBD.

Administration, Oral↗
PubMed2001

[Opportunistic pneumonia after kidney transplantation].

This study was conducted to evaluate clinical features at the onset of pneumonia and the usefulness of methods for diagnosing pneumonia in patients who have undergone kidney transplantation. From January 1990 to December 1998. 174 kidney transplantations were performed, and were followed by 22 cases of pneumonia. Of the 22 pneumonia patients, 16 were male and 6 were female. The median age of the 22 patients was 37.2 +/- 13.3 years. All the patients received cyclosporin A and corticosteroids. In 11 cases, the organisms were identified in the microbiology or pathology laboratory, either during life or at autopsy. Six cases were due to Pneumocystis carinii (PC), three to PC and Cytomegalovirus (CMV), one to Aspergillus, and one resulted from miliary tuberculosis. Pneumonia occurred within 4 months after kidney transplantation in most cases. The mean interval between the transplantation and the appearance of pneumonia was 77.3 +/- 34.3 days, except in the cases of Aspergillosis and miliary tuberculosis, where the intervals were 46 and 50 months, respectively. The mean interval from the appearance of symptoms to the detection of pulmonary infiltration was 3.3 +/- 4.3 days. The clinical features present when pulmonary infiltration was detected by CT were fever (91%), cough (32%), and crackles (27%). However, at this time, 55% of the cases had no symptoms other than fever. Chest radiographs were positive for pulmonary infiltration in 64% of the cases at the same time that the pulmonary infiltrates were detected by CT. Eighty percent of the cases exhibited diffuse interstitial infiltrates. Organisms were detected in 7 of 9 cases examined with bronchofiberscopy (BF). But in only one of 13 cases that did not undergo BF. Increased values of serum beta-D-glucan were detected in the early phase of three PC pneumonia cases, suggesting that beta-D-glucan is useful as a marker of PC pneumonia. The use of bronchofiberscopy was more frequent in survivors of PC pneumonia than in non-survivors, whereas the mean age was higher and coexisting CMV infections were identified more frequently in the non-survivors. We concluded that fever is important as an initial symptom of pulmonary infection. In addition, we find that CT is very useful for the detection of interstitial infiltrates, and BF is an excellent method for detecting organisms in the pneumonia patient after kidney transplantation.

Adult↗
PubMed2001

Immunochemical detection of a lipofuscin-like fluorophore derived from malondialdehyde and lysine.

The accumulation of fluorescent age pigment or lipofuscin is a frequently observed age-associated cellular alteration in a variety of postmitotic cells of many species. These pigments are observed within granules composed, in part, of damaged protein and lipid. Modification of various biomolecules by aldehyde products of lipid peroxidation is believed to contribute to lipofuscin and ceroid formation. In the present study, we raised a monoclonal antibody (MAb 1F83) directed to the malondialdehyde-modified protein and identified a lipofuscin-like dihydropyridine fluorophore as the major epitope. This antibody was used to conclusively demonstrate that the fluorophore forms on oxidatively modified low density lipoproteins. In addition, we demonstrated that the materials immunoreactive to MAb 1F83 indeed constituted the atherosclerotic lesions, in which intense positivity was associated primarily with macrophage-derived foam cells. The results of this study suggest that the reaction between the lipid peroxidation-derived aldehyde and primary amino groups of protein might represent a process common to the formation of the lipofuscin-like fluorophore during aging and its related diseases.

Animals↗
PubMed2001

[Therapeutic efficacy of a topical tolnaftate preparation in guinea pig model of tinea pedis].

The therapeutic efficacy of a topical antifungal ointment containing 2% tolnaftate was studied in a guinea pig model of tinea pedis using the following four topical antifungal preparations commercially available as reference drugs: variotin (3,000 U/g ointment); phenyl-11-iodo-10-undecynoate (0.5% ointment); siccanin (1% ointment); and clotrimazole (1% cream). After the infection fully developed, the infected animals were treated twice daily with the testing drug or reference drug for consecutive four weeks. Therapeutic efficacy was mainly evaluated on the basis of the extent of the yield of fungal cultures from the infected skin tissues (infection intensity) at the end of the treatment period. In animal groups treated with tolnaftate as well as with three reference drugs, siccanin, phenyl-11-iodo-10-undecynoate and clotrimazole, the average infection intensity was significantly lower than that for untreated control group (P < 0.05-0.005) although no culture-negative animal was seen in any treated animal group. Comparing with all the reference drugs, tolnaftate was the most highly effective and there was significant difference in the average infection intensity between a tolnaftate treated and each reference drug-treated groups (P < 0.01-0.005). These results confirm the clinical usefulness of the current tolnaftate preparation in the treatment of patients with tinea pedis and probably other clinical forms of dermatophytoses.

Administration, Topical↗
PubMed2001

[A case of transitional cell carcinoma of the urinary bladder with high serum level of CEA and CA19-9].

An 85-year-old male with asymptomatic gross hematuria was diagnosed with invasive bladder tumor, transitional cell carcinoma grade 3. Serum levels of CEA and CA19-9 were elevated and histological examination revealed expression of both markers in the cytoplasm of cancer cells. Out of therapeutic options, intra-arterial chemotherapy and radiotherapy were selected because of his age. During the treatment, serum levels of CEA and CA19-9 decreased along with reduction of tumor size. These serum markers have been reported to be elevated in 10 to 60% of patients with bladder tumor and are useful markers for evaluation of the treatment as suggested in the present case.

Aged↗
PubMed2001

[Prostate cancer].

Prostate specific antigen (PSA) has been widely used as a tumor marker for screening, diagnosis and monitoring of prostate cancer. Use of PSA is very important for diagnosis during the primary screening for the early detection of prostate cancer. The presence of various PSA assays and with no uniform serum data have led to confusion among clinicians in the evaluation of serum data. The characteristics of each PSA assay, such as the equimolar or skewed type, must be taken into consideration at the time of use. For the early detection of prostate cancer, PSA density (PSAD), PSA adjusted for the transiton zone volume (PSATZ) and free to total PSA ratio may be useful in the selection of patients with intermediate PSA levels who should undergo prostate needle biopsy.

Adult↗
PubMed2001

Antitumor effects of the intravesical instillation of heat killed cells of the Lactobacillus casei strain Shirota on the murine orthotopic bladder tumor MBT-2.

PURPOSE: To characterize the potential of heat killed Lactobacillus casei, Shirota strain (LC9018), as an alternative to bacillus Calmette-Guerin (BCG) for treating patients with bladder cancer we investigated the antitumor effects of intravesical instillation of LC9018 in the MBT-2 orthotopic bladder tumor implantation model in C3H/He mice. MATERIALS AND METHODS: LC9018 or BCG, Tokyo 172 strain, was instilled once daily for 10 days starting on the day after orthotopic implantation of MBT-2. Tumor appearance and mean bladder weight on day 21 after tumor implantation were evaluated. Moreover, we investigated the augmentation of local cellular immunity in bladder mucosa by immunohistochemical staining and reverse transcription polymerase chain reaction. RESULTS: Intravesical LC9018 instillation significantly reduced the rate of tumor appearance in 8 of 38 subjects (p <0.001) and mean tumor growth plus or minus standard deviation with a bladder weight of 37 +/- 49 mg. (p <0.001) compared with tumor appearance in 41 of 58 subjects and mean bladder weight 146 +/- 183 mg. in controls. BCG had no significant antitumor activity in the orthotopic implantation model. Intravesical instillation of LC9018 augmented the local expression of antitumor cytokine messenger RNA (interferon-gamma and tumor necrosis factor-alpha) and induced the infiltration of neutrophils surrounded by macrophages that phagocytosed LC9018 cells at the bladder mucosa. CONCLUSIONS: These results suggest that LC9018 is potentially more potent and safer as a therapeutic agent than BCG for superficial bladder tumors. Furthermore, the antitumor effect of LC9018 is exerted via the augmentation of local cell mediated antitumor immunity.

Administration, Intravesical↗
PubMed2001

[Breast cancer].

Neoadjuvant chemotherapy (NAC) represents a new approach based on sound theoretical, pharmacokinetic, and experimental principles. The purpose of NAC is to improve control of the primary site by downstaging and to improve control of micrometastatic disease. NAC has been standard therapy in the management of locally advanced breast cancer. Patients with earlier stage breast cancer may also benefit from treatment with NAC. Recently some investigators have mentioned that NAC can be used instead of adjuvant chemotherapy and would be most appropriate for patients who wish to preserve their breast but who have tumors too large for breast conserving surgery. In this article, we reviewed the present status of NAC (indication, clinical response, pathologic response, survival, possibility of breast conservation, prognostic/predictive factors, neoadjuvant endocrine therapy) and discussed several unanswered questions on NAC (survival benefit, optimal number of treatment cycles, optimal regimens) and future direction. Combined modality therapy including NAC appears to provide excellent local control, the possibility of breast conservation, and, probably, an increased survival rate, at least for some subsets of patients. Furthermore, through sequential sampling, NAC provides indeed the opportunity to identify molecular mechanisms associated with pathologic response and to study the possibility to guide the choice for induction treatment and patient populations submitted to neoadjuvant chemotherapy.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed2001

Tubular osteopontin expression in patients with ANCA-associated glomerulonephritis.

OBJECTIVE: To elucidate the role of osteopontin (OPN) in monocyte recruitment in crescentic glomerulonephritis, we investigated immunohistochemical localization of OPN in the kidney and its correlation with clinical and histopathologic parameters in biopsy specimens of patients with myeloperoxidase antineutrophil cytoplasmic autoantibody- (MPO-ANCA) associated glomerulonephritis. METHODS: Twelve patients with MPO-ANCA-associated glomerulonephritis were enrolled in this study. Clinical parameters such as creatinine clearance and urinary protein excretion of each patient were obtained at the time of biopsy. Paraffin-embedded sections were used for immunohistochemical staining using the LSAB method. Five cortical interstitial fields randomly selected at original magnification x 200 were assessed using a computer-assisted color image analyzer. Tubular OPN expression was assessed as the percentage of positive area in the tubulointerstitium. Double immunofluorescent staining using antibodies against OPN and alpha(v)beta3 was performed. RESULTS: In all of the cases studied, OPN was occasionally localized within the glomeruli, and expressed slightly in proximal tubular epithelium and significantly in distal tubular epithelium. Tubular OPN expression tended to be promoted in the interstitium infiltrating by numerous monocytes/macrophages. The extent of tubular OPN expression was positively correlated with serum ANCA titers and urinary OPN concentrations. Enhanced alpha(v)beta3 expression appeared in the distal tubular epithelium expressing OPN. CONCLUSION: These results suggest that inducible expression of OPN and alpha(v)beta3 in the tubular epithelium seems to be associated with interstitial moncyte infiltration and subsequent tubulointerstitial changes in human MPO-ANCA-associated glomerulonephritis.

Adult↗
PubMed2001

2-Aminoethoxydiphenyl borate (2-APB) inhibits capacitative calcium entry independently of the function of inositol 1,4,5-trisphosphate receptors.

Capacitative calcium entry (CCE), the mechanism that replenishes intracellular calcium stores after depletion, is essential to intracellular calcium signaling. CCE is mediated by the channels in the plasma membrane generally referred to as "store operated channels (SOCs)". However, the molecular identity of the SOCs has never been determined, and the mechanism of the activation of SOCs remains to be elucidated. Recent studies have demonstrated that 2-aminoethoxydiphenyl borate (2-APB), which has been found to be an antagonist of inositol 1,4,5-trisphosphate receptors (IP3Rs), inhibits CCE, suggesting that IP3Rs channel activity is essential to the generation of CCE. However, CCE has also been reported to occur normally in IP3R-deficient cells. In order to resolve this discrepancy, we investigated the effect of 2-APB on CCE in IP3Rs-deficient cells. In response to store depletion with thapsigargin or N,N,N',N'-tetrakis (2-pyridylmethyl) ethylene diamine (TPEN), CCE was generated in IP3Rs-deficient cells the same as in wild-type cells, however, 2-APB abolished CCE in IP3Rs-deficient cells, despite the fact that this cell line does not possess functional IP3Rs. We also examined the effect of 2-APB on several types of TRP Ca2+ channels, which exhibit properties similar to those of SOCs. 2-APB had a different inhibitory effect on spontaneous and thapsigargin-induced Ba2+ influx in cells that transiently expressed individual TRP subtypes. These results suggest that the channel activity of IP3Rs is not essential to the generation of CCE in this cell line and that 2-APB inhibits CCE independently of the function of IP3Rs.

Boron Compounds↗