Biomedical subjects

H Sakahara

Publications and source records attributed to H Sakahara.

At least 73 records · Page 4Linked to original sources

PubMed1997

Intravenous avidin chase improved localization of radiolabeled streptavidin in intraperitoneal xenograft pretargeted with biotinylated antibody.

In the present study, we examined the effect of avidin administered intravenously (i.v.) on the biodistribution of radiolabeled streptavidin in mice bearing intraperitoneal (IP) xenografts pretargeted with biotinylated antibody. Tumors were established in nude mice by IP inoculation of LS180 human colon cancer cells. Monoclonal antibody MLS128, which recognizes Tn antigen on mucin, was biotinylated and injected IP into the IP tumor-bearing mice. Radioiodinated streptavidin was administered IP or i.v. 48 h after pretargeting of biotinylated antibody. Avidin was administered i.v. 30 min prior to streptavidin injection. The localization of radioiodinated streptavidin in the tumor pretargeted with biotinylated antibody was significantly higher than that without pretargeting and that of radioiodinated MLS128 by the one-step method. Avidin administration significantly accelerated the clearance of radioiodinated streptavidin in blood and other normal tissues and increased the tumor-to-blood radioactivity ratio regardless of administration route of streptavidin. The i.v. avidin chase improved tumor localization of radiolabeled streptavidin in the IP xenografts pretargeted with biotinylated antibody.

Animals↗

PubMed1997

Radioiodinated alpha-p-iodophenyl-N-tert-butylnitrone as a radical detecting agent in vivo.

For in vivo detection of unstable radical species, a novel radioiodinated radical trapping agent, alpha-p-iodophenyl-N-tert-butylnitrone (IPBN), was designed based on the plausible characteristics of alpha-p-hydroxy-m-iodophenyl-N-tert-butylnitrone (HIPBN). Both compounds are iodinated analogs of alpha-phenyl-N-tert-butylnitrone (PBN), but the latter showed low stability in vivo. The present compound IPBN was easily prepared and labeled with radioiodine. It showed very stable characteristics in vitro as well as in vivo, but was easily metabolized and retained at sites of superoxide production.

Animals↗

PubMed1997

Expression of vascular endothelial growth factor (VEGF) in epithelial ovarian neoplasms: correlation with clinicopathology and patient survival, and analysis of serum VEGF levels.

Vascular endothelial growth factor (VEGF) is known to be produced by various solid tumours and is thought to be involved in microvascular permeability and/or angiogenesis. To examine the relationship between VEGF expression in ovarian neoplasms and clinicopathological factors or patient survival, expression of VEGF was analysed immunohistochemically in 110 epithelial ovarian tumours. In addition, VEGF levels in the tumour fluid (17 patients), ascites (12 patients) and sera (38 patients) were determined using enzyme immunoassay. Positive immunostaining for VEGF was observed in 97% (68 out of 70) of ovarian carcinomas, which was significantly higher than that of tumours of low malignant potential (LMP) (13 out of 25; 52%) and benign cystadenomas (5 out of 15; 33%) (P < 0.01). In ovarian carcinomas, strong VEGF immunostaining was also observed more frequently in tumours of clear cell type (P < 0.05) in the advanced stage of disease (P < 0.05) and with positive peritoneal cytology (P < 0.01). Patients with strong VEGF staining had poorer survival rates than those with weak or no immunostaining for VEGF (P < 0.01). These findings suggest that strong VEGF expression plays an important role in the tumour progression of ovarian carcinoma. The enzyme immunoassay revealed higher serum VEGF levels in carcinoma patients than those in patients with LMP or benign tumours (P < 0.01). Serum VEGF levels decreased after the successful removal of tumours in ovarian cancer patients and, in one patient, the serum VEGF level was re-elevated during relapse. Therefore, serum VEGF could be used as a marker for monitoring the clinical course of ovarian cancer patients.

Adult↗

PubMed1997

An in vitro 1H-MRS model of oncogene transfection. The spectral feature of c-erbB-2 and c-Ha-ras transfected NIH3T3 fibroblast cells.

PURPOSE: Malignancy is an abnormality of cell division and differentiation based on abnormal expression of oncogenes. This note describes the in vitro 1H-NMR spectral features of oncogene-transfected NIH3T3 fibroblast cells compared to non-transfected cells. MATERIAL AND METHODS: 1H-NMR spectra of cultured NIH3T3 cells and c-erbB-2 or c-Ha-ras gene-transfected cells were obtained by 400 MHz high resolution NMR. The peaks were assigned by 2D HOHAHA spectra of the cell suspension and the spectral changes were evaluated in 1D and 1D differential spectra. RESULTS: The 1H spectra obtained from both transfected cell lines were broadened over all peaks, suggesting reduced mobility in plasma membrane lipid molecules. No other differential spectra for characterizing metabolic change was detected. CONCLUSION: Broadened 1H spectra observed after c-erbB-2 or c-Ha-ras transfection suggest changes of plasma membrane viscosity, which may be related to the oncogene expression.

3T3 Cells↗

PubMed1997

Noninvasive quantitative measurements of regional cerebral blood flow using technetium-99m-L,L-ECD SPECT activated with acetazolamide: quantification analysis by equal-volume-split 99mTc-ECD consecutive SPECT method.

Resting- and acetazolamide (Acz)-activated-regional cerebral blood flow (rCBF) measurements were performed by consecutive single-photon emission computed tomography (SPECT) studies before and after Acz administration using equal-volume-split technetium-99m-L,L-ethyl cysteinate dimer. Quantitative rCBF images were converted from qualitative axial SPECT images by the application of Patlak plot graphical analysis with radionuclide angiography and Lassen's linearization correction. Total time span required for this study was 53 minutes. The unaffected side of 37 studies with unilateral vascular lesions and 45 studies without apparent vascular lesions showed 132 +/- 17% and 140 +/- 15% increase of mean CBF (mCBF), respectively, under Acz administration. Comparing these values, the Acz-activated rCBF increases of less-affected and affected hemispheres of 23 studies with bilateral vascular lesions (116 +/- 13% and 113 +/- 12%, respectively) was lower with high statistical significance (P < 0.001). For the other 20 cases, physiologic saline was administered instead of Acz. This group showed no changes in mCBF under placebo administration (after placebo/baseline; 100 +/- 6%). Acetazolamide-activated rCBF increase was recognized clearly and easily using quantitative images. This noninvasive method is easy to perform and may be helpful to detect regional abnormalities of hemodynamic reserve in cerebrovascular diseases.

Acetazolamide↗

PubMed1997

Detection of altered adhesion molecule expression in experimental tumors by a radiolabeled monoclonal antibody.

Adhesion molecules play a major role in the processes of invasion and metastasis of malignant tumors. Their expression within tumors has been reported to be quantitatively and qualitatively altered according to the invasiveness and metastatic potential of the tumor. The present study tested whether the intratumoral expression of integrin alpha 3 can be detected by a radiolabeled monoclonal antibody. The in vitro binding study with four different human cancer cells showed that radioiodinated GA17 antibody recognizing integrin alpha 3 bound specifically to these cells to varying degrees, according to the antigen density on each cell. The biodistribution study with 125I- and 111In-labeled antibodies showed specific localization of radiolabeled GA17 to the xenografts. However, the in vivo tumor localization was not proportional to the antigen density calculated in vitro, and antibody metabolism varied among the tumors, as was also confirmed by in vitro radionuclide retention assay. The intratumoral distribution of radioactivities varied reflecting the antigen expression within the tumor. These results indicate that 1) integrin alpha 3 was expressed in various kinds of tumors and could be localized by the radiolabeled antibody, and 2) the expression of integrin alpha 3 and the metabolism of the radiolabeled antibody after binding to the antigen within the tumor were variable among the tumors, which affected the radionuclide distribution characteristics. The expression of adhesion molecules within these tumors was noninvasively detected by a radiolabeled antibody. It may be possible to use integrin alpha 3, when it is overexpressed, as a target of therapy with antibodies radiolabeled with alpha or beta emitters.

Animals↗

PubMed1997

Anti-murine antibody response to mouse monoclonal antibodies in cancer patients.

Although development of human anti-murine immunoglobulin antibody (HAMA) is often seen in patients receiving murine antibodies, the variety of methods used for detecting HAMA makes it difficult to compare directly the HAMA responses measured by different assays. In the present study, several parameters of the HAMA response to two murine monoclonal antibodies were evaluated. The anti-sialosyl Tn antibody MLS102 and anti-CA125 antibody 145-9, which were labeled with 111In, were injected intravenously into 17 colorectal cancer patients and 11 ovarian cancer patients for immunoscintigraphy, respectively. HAMA was measured by enzyme-linked immunosorbent assay. There was no difference in baseline HAMA levels before antibody injection between the two groups. HAMA developed more frequently in ovarian cancer patients receiving the 145-9 antibody than in colorectal cancer patients receiving the MLS102 antibody (9/11 vs. 6/17, P < 0.05). No significant difference was observed in maximal HAMA levels between the two groups of patients. However, time to reach the maximal levels was delayed and the duration of the response seemed longer in ovarian cancer patients. Among 11 patients receiving the 145-9 antibody three patients became positive for HAMA more than 2 months after antibody injection and the other two had HAMA activity in their sera for more than 17 months. HAMA response was different between the two antibodies, and late onset or long duration of HAMA response against the 145-9 antibody suggests the importance of HAMA measurement in patients who receive a second injection of murine antibodies even after a long interval.

Adult↗

PubMed1997

Three phase 99Tcm (V)DMSA scintigraphy in Paget's disease: an indicator of pamidronate effect.

The use of three phase 99Tcm (V)DMSA scintigraphy is reported in a patient with Paget's disease of bone before and after intravenous pamidronate therapy. It was a useful modality for estimating the activity of Pagetoid lesions and the therapeutic effect of pamidronate, from a different aspect to bone scintigraphy. Three phase 99Tcm (V)DMSA scintigraphy evaluates both the blood flow and the metabolic activity of Pagetoid bone.

Aged↗

PubMed1997

Transient increase in glycolytic metabolism in cultured tumor cells immediately after exposure to ionizing radiation: from gene expression to deoxyglucose uptake.

The transient change in uptake of deoxyglucose (DG) and expression of glycolysis-associated gene products in cultured tumor cells (LS180 human colon adenocarcinoma cells) immediately after single-dose X irradiation were examined to acquire basic data for use in the early assessment of tumor responses to radiation treatment by position emission tomography. An increase in accumulation of DG was found 3-5 h postirradiation. Inhibitors of both mRNA and protein synthesis and glycoprotein transport suppressed the increase in accumulation of DG to the control level. Both the glucose transporter-1 mRNA expression and the enzymatic activity of hexokinase in the cells were significantly elevated in conjunction with high DG accumulation. These findings indicate that the transiently elevated glucose metabolism occurred via processes at the levels of gene expression. These transient tumor cell responses might be useful for the early assessment of radiation damage.

Colonic Neoplasms↗

PubMed1997

[Evaluation of clinical utility of 111In-DTPA-IgG scintigraphy in the detection of inflammation/infection--a report of multicenter phase III clinical trials].

This is the report of phase III study to evaluate the safety and utility of 111In-DTPA-IgG in patients with strongly suspected inflammation/infection. One hundred and forty five patients with suspected sites of inflammation/infection were enrolled in the study. Only a few adverse experiences in one patient were reported, which were interpreted as having a possible relationship to the agent. A total of 171 suspected sites (12 in head and neck, 39 in thorax, 44 in abdomen and pelvis, 62 in musculoskeletal system, and 14 in other regions) were evaluated by investigators at each institute. Out of 171 sites, 18 were determined to be unevaluable, and 12 false negative and 5 false positive cases were observed. Overall sensitivity and specificity was 89.8% and 85.7%, respectively, and the agent proved to be effective in detecting lesions anywhere throughout the body. The analysis of data from this Phase III study indicates that 111In-DTPA-IgG is well-tolerated in patients and effective in determining focal sites of inflammation/infection although the physiological accumulation in some tissues such as the sinus or liver and possible excretion into the gastrointestinal tract may make it difficult to localize lesions.

Aged↗

PubMed1996

Reassessment of diethylenetriaminepentaacetic acid (DTPA) as a chelating agent for indium-111 labeling of polypeptides using a newly synthesized monoreactive DTPA derivative.

Previous studies on indium-111 (111In) labeling of polypeptides and peptides using cyclic diethylenetriaminepentaacetic dianhydride (cDTPA) as a bifunctional chelating agent (BCA) have indicated that DTPA might be a useful BCA for 111In labeling of polypeptides at high specific activities when DTPA can be incorporated without inducing intra- or intermolecular cross-linking. To investigate this hypothesis, a monoreactive DTPA derivative with a maleimide group as the peptide binding site (MDTPA) was designed and synthesized. A monoclonal antibody (OST7, IgG1) was used as a model polypeptide, and conjugation of MDTPA with OST7, 111In radiolabeling of MDTPA-OST7, and the stability of 111In-MDTPA-OST7 were investigated using cDTPA and benzyl-EDTA derivatives as references. SDS-PAGE analysis demonstrated that while cDTPA induced intramolecular cross-linking, no such undesirable side reactions were observed with MDTPA. MDTPA generated 111In-labeled OST7 with high radiochemical yields at higher specific activities than those produced using cDTPA and benzyl-EDTA derivatives as the BCAs. Incubation of each 111In-labeled OST7 in human serum indicated that MDTPA generated 111In-labeled OST7 of much higher and a little lower stability than those derived from cDTPA and benzyl-EDTA derivatives, respectively. These findings indicated that the low in vivo stability of cDTPA-conjugated antibody reported previously is not attributable to low stability of 111In-DTPA but to formation of intramolecular cross-linking during cDTPA conjugation reactions. The present study also indicated that MDTPA and its precursor, the tetra-tert-butyl derivative of DTPA, would be useful BCAs for 111In radiolabeling of polypeptides that have rapid blood clearance with high specific activities.

Chelating Agents↗

PubMed1996

Sonographic characteristics of recurrent hepatocellular carcinoma.

Our purpose was to determine the characteristics findings of recurrent hepatocellular carcinomas (HCCs) on sonography. We reviewed sonographic findings of 85 recurrent tumors in 57 patients with prior partial hepatectomy for HCCs. Of 57 patients, 47 (82.5%) had localized recurrence; one or two tumors in a segment (44 patients) and a recurrent tumor near the resected hepatic stump (3 patients). Of 85 tumors, 58 (68.2%) were less than 2 cm, and 82 of 85 tumors (96.5%) were less than 4 cm. A total of 49 tumors (57.6%) were seen as hypoechoic, and 20 tumors (23.5%) as hyperechoic. Posterior echo enhancement was seen in 36 tumors (42.4%), hypoechoic halo in 24 tumors (28.2%), lateral shadow in 21 tumors (24.7%), and mosaic pattern in 6 tumors (7.1%). Recurrent HCC can be detected at an early stage by sonography. Hypoechoic pattern and posterior echo enhancement are commonly seen. These findings suggest recurrent HCCs.

Aged↗

PubMed1996

Periosteal ganglion of the tibia.

Radiographic, computed tomographic and scintigraphic findings of a patient with separated, multilocular periosteal ganglion are reported. Multiple periosteal cystic masses with calcification in small parts of the cyst walls were demonstrated in the surface of the left tibia by plain radiograms and CT. The accumulations of technetium-99m hydroxymethylene diphosphonate and pentavalent technetium-99m dimercaptosuccinic acid were shown in the calcification in the periosteum and wall of the cyst. Needle puncture revealed that the masses were filled with jelly-like fluid. The masses were diagnosed as multiple ganglionic cysts at the periosteum of the left tibia.

Aged↗

PubMed1996

Inflammation-seeking scintigraphy with radiolabeled biotinylated polyclonal IgG followed by the injection of avidin chase.

We tried to apply the avidin chasing system to the inflammation-seeking scintigraphy using radiolabeled nonspecific polyclonal IgG. We studied the pharmacokinetics of technetium-99m and iodine-125-labeled biotinylated murine polyclonal IgG followed by an avidin chase injection in model mice with inflammatory foci. Avidin chase decreased the circulating radioactivity of 99mTc and 125I, which was a major problem for inflammation-seeking scintigraphy using radiolabeled nonspecific polyclonal IgG, to 9.3% and 19.3% of that without an avidin chase injection, respectively. Inflammation-seeking scintigraphy with the aforementioned method would be better than that with conventional method.

Animals↗

PubMed1996

Stability of a metabolizable ester bond in radioimmunoconjugates.

Ester bonds have been used as metabolizable linkages to reduce radioactivity levels in non-target tissues following the administration of antibodies labeled with metallic radionuclides. In this radiochemical design of antibodies, while the ester bonds should be cleaved rapidly in non-target tissues, high stability of the ester bonds in plasma is also required to preserve target radioactivity levels. To assess the structural requirements to stabilize the ester bond, a new benzyl-EDTA-derived bifunctional chelating agent with an ester bond, (1-[4-[4-(2- maleimidoethoxy)succinamido]benzyl]ethylenediamine-N,N,N',N' -tetraacetic acid; MESS-Bz-EDTA), was developed. MESS-Bz-EDTA was coupled with a thiolated monoclonal antibody (OST7, IgG1) prepared by reducing its disulfide bonds to introduce the ester bond close and proximal to the antibody molecule. For comparison, 1-[4-(5- maleimidopentyl)aminobenzyl]ethylenediamine-N,N,N',N'-tetraacetic acid (EMCS-Bz-EDTA) and meleimidoethyl 3-[131I]iodohippurate (MIH) was coupled to OST7 under the same conjunction chemistry. When incubated in 50% murine plasma or a buffered-solution of neutral pH, OST7-MESS-Bz-EDTA-111In rapidly released the radioactivity, and more than 95% of the initial radioactivity was liberated after a 24 h incubation in both solutions, due to a cleavage of the ester bond. On the other hand, only about 20% of the radioactivity was released from OST7-MIH-131I in both solutions during the same incubation period. In mice biodistribution studies, while a slightly faster radioactivity clearance from the blood with less radioactivity levels in the liver and kidneys was observed with OST7-MIH-131I than with OST7-EMCS-Bz-EDTA-111In, OST7-MESS-Bz-EDTA-111In indicated radioactivity clearance from the blood much faster than and almost comparable to that of OST7-MIH-131I and succinamidobenzyl-EDTA-111In, respectively. These findings as well as previous findings on radiolabeled antibodies with ester bonds suggested that while an introduction of an ester bond close to an antibody molecule stabilized the ester bond against esterase access, chemical structures of the linkages and radiolabels attached to the ester bonds play a significant role in the chemical stability of the ester bond. This may explain the different stability of the ester bonds in radioimmunoconjugates so far reported.

Animals↗

PubMed1996

1H-magnetic resonance spectroscopic observation of cultured malignant cells pharmacologically induced to different phenotypes.

RATIONALE AND OBJECTIVES: We evaluated the 1H nuclear magnetic resonance spectra of malignant cells after the administration of drugs that cause morphologic changes. METHODS: 1H spectra of a human lung adenocarcinoma cell line cultured with interferon gamma, dexamethasone, or sodium butyrate were obtained. The peaks were assigned by two-dimensional homonuclear Hartmann-Hahn spectroscopy spectra of the cells and their perchloric acid extracts. Differential spectra were used to evaluate relative changes in the peaks. RESULTS: In the control culture, choline/phosphocholine peaks were increased in the cell-growth phase, and the 1.26-ppm peak was increased in the confluent state. Treatment by interferon gamma and dexamethasone induced reproducible changes in the peaks of differential spectra corresponding to 1.26 ppm, choline/phosphocholine, and glutamate/glutamine. Dexamethasone treatment broadened lipid peaks. Changes after treatment with sodium butyrate were obscure. Microscopically, cells were induced to morphologically different phenotypes by each drug. CONCLUSION: Cells induced to exhibit morphologically different phenotypes present different 1H spectra.

Adenocarcinoma↗

PubMed1996

Effect of circulating antigen on immunoscintigraphy of ovarian cancer patients using anti-CA125 monoclonal antibody.

The murine monoclonal antibody (mAb) 145-9 recognizes an epitope present on CA125 but different from the epitope defined by the mAb OC125. To evaluate the clinical usefulness of the 145-9 antibody, immunoscintigraphy was performed in ovarian cancer patients and the effect of circulating CA125 on tumor imaging was investigated. Two milligrams (74 MBq) of 111In-labeled 145-9 was injected intravenously into 11 patients with ovarian cancer. Pre-injection serum CA125 concentrations were between 166 U/ml and 7414 U/ml. Tumors were visualized in 10 of 11 patients. In two patients, lymph nodes that were not detected by other imaging modalities but were clinically suspected as metastases were visualized. There was no correlation between serum CA125 level and antibody uptake in the tumors. Immune complexes between the antibody and circulating antigen were observed in sera of all the patients, but the fraction of radioactivity in complex form did not correlate well with serum CA125 levels. The immune complexes survived in the circulation and the circulating radiolabel, including immune complexes, was still bound to solid-phase CA125. The plasma clearance rate and hepatic uptake of the antibody were not significantly affected by circulating CA125. In conclusion, the antibody 145-9 formed complexes with CA125 in vivo but this did not Compromise the outcome of antibody imaging. The antibody 145-9 can be used in immunoscintigraphy of ovarian cancer irrespective of serum CA125 level.

Adult↗

PubMed1996

Specificity and affinity of 26 monoclonal antibodies against the CA 125 antigen: first report from the ISOBM TD-1 workshop. International Society for Oncodevelopmental Biology and Medicine.

The specificity of 26 monoclonal antibodies against the CA 125 antigen was investigated in two phases of the ISOBM TD-1 workshop. The binding specificity was studied using CA 125 immunoextracted by specific antibodies immobilized on various solid phases, or on the surface of human cell lines. Immunometric assays using all possible antibody combinations were used to study the topography of antibody binding sites on the antigen. We conclude that the CA 125 antigen carries only two major antigenic domains, which classifies the antibodies as OC125-like (group A) or M11-like (group B). One antibody, OV 197, showed binding specificity related to some of the OC125-like antibodies, but was classified into a separate group C. The OC125-like group of antibodies has four subgroups with different binding specificities. These are A1 = OC 125 and K 95, A2 = K 93, A3 = B43.13, and A4 = ZS 33, B27.1 and CCD 247. Binding of nonlabelled OC 125 or K 95 to CA 125 caused a marked increase in binding of labelled OV 197 to the complex. This conformational change was not observed with any other antibody combinations. Antibody B43.13 could form immunometric assay combinations particularly with antibodies of subgroup A4, indicating that the B43.13 epitope is in the periphery of the binding area of OC125-like antibodies. The M11-like group of antibodies is more homogenous with strong cross-inhibition between most antibodies. Only one antibody, ZR 38, would form an immunoassay combination with other M11-like antibodies and thus represents a distinct subgroup. The main group of M11-like antibodies are M 11, ZR 45, MA602-6, K 91, OV 185, K 101, K 90, K 96, K 97, K 102, CCD 242, 145-9, and 130-22. Antibody OV 197 binds to a domain designated C and is unique, as stated above. Antibody pairs from any two of the three groups may be used in immunometric assays. Three antibodies were not studied by complete cross-inhibition due to low affinity (OV 198 and K 100) or lack of material (MA602-1). OV 198 and K 100 are most likely OC125-like and MA602-1 is M11-like. Antibody affinity was estimated with labelled antigen in solution or with antigen absorbed on microtiter wells. Western blot analysis showed staining both in the stacking gel and corresponding to a molecule of 200 kDa. There was a marked difference between the antibodies in their ability to bind to CA 125 immobilized on a membrane. Strongest binding was observed with the M11-like antibodies, particularly M 11, K 96, K 97, MA602-6, 145-9. Antibodies belonging to the subgroup A4 were the only OC 125-like antibodies which reacted well with CA 125 in Western analysis. Digestion of CA 125 with proteolytic enzymes showed it to be particularly sensitive to trypsin cleavage. However, no low molecular weight fragments with preserved immunoreactivity were found.

Animals↗