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Biomedical subjects

S Ueda

Publications and source records attributed to S Ueda.

At least 217 records · Page 12Linked to original sources

Expression and purification of biologically active porcine follicle-stimulating hormone in insect cells bearing a baculovirus vector.

Biologically active recombinant porcine FSH (rec-pFSH) free from the cognate pituitary glycoprotein hormone LH was produced. It was synthesized by a baculovirus vector-insect cell system using two cDNAs encoding the glycoprotein alpha and FSH beta subunits. Its antigenicity was the same as that of pFSH prepared from the pituitary. Glycosylation of rec-pFSH was shown by tunicamycin treatment but the molecular mass of each subunit was lower than that of pituitary-derived FSH, because of the absence of trimming of terminal sugars in insect cells. Rec-pFSH was secreted into the culture medium at about 1 mg/l and purified in six fractions, because of the heterogeneity of the sugar group, by S-Sepharose and concanavalin A-Sepharose column chromatography. The biological activity of rec-pFSH was examined by measuring its effect on progesterone secretion from porcine granulosa cells and germinal vesicle breakdown (GVBD) of porcine oocytes. It showed adequate activity with respect to progesterone secretion, although some fractions rich in the sugar group showed lower activity than that of pituitary-derived FSH. It exhibited higher GVBD activity than that of pituitary-derived FSH at concentrations as low as 1 ng/ml. These results demonstrate that the baculovirus vector-insect cell system can provide biologically active rec-pFSH.

Animals↗

Hyperperfusion syndrome following carotid endarterectomy: evaluation using diffusion-weighted magnetic resonance imaging--case report.

A 65-year-old female developed hyperperfusion syndrome following carotid endarterectomy for severe stenosis of the left internal carotid artery. Transcranial Doppler monitoring showed a sharp increase in flow velocity in the middle cerebral artery (MCA). Diffusion-weighted magnetic resonance (MR) imaging demonstrated diffuse hyperintensity in the region of the left MCA, which diminished markedly 9 days later. The abnormal hyperintensity decreased as the patient's symptoms improved, suggesting that diffusion-weighted MR imaging can be used for evaluating the course of hyperperfusion syndrome.

Aged↗

Determination of glycated albumin by enzyme-linked boronate immunoassay (ELBIA).

A new affinity method for quantification of glycated albumin by an enzyme-linked boronate-immunoassay (ELBIA) has been established, based on the interaction between boronic acids and the cis-diols of glycated human serum albumin (HSA) trapped by anti-HSA antibody. To evaluate the ELBIA, we first examined the accuracy of the conventional boronate affinity chromatographic (BAC) method. In the BAC method, 8.1-18.9% of nonglycated albumin calibrator nonspecifically bound to the boronate affinity column, values that were regarded as the column blank. In the modified BAC method, therefore, we substracted the column blank value from the measured glycated albumin value to obtain the true value. Because glycated albumin values measured by ELBIA were exactly the same as reported by the modified BAC method, we suggest that the ELBIA results reflect the real status of albumin glycation. We have also developed a fully automated ELBIA system, allowing multiple, rapid, and precise measurements of glycated albumin.

Boronic Acids↗

Fluorine-18-labeled fluoroboronophenylalanine PET in patients with glioma.

UNLABELLED: We synthesized fluorine-18-labeled fluoroboronophenylalanine (18F-10B-FBPA), an analog of boronophenylalanine (10B-BPA), and characterized its pharmacokinetics in patients with glioma. We conducted PET studies on three types of gliomas to clarify the relationship between tumor grade and each rate constant [K1 (ml/g/min), k2 (min[-1]) and k3 (min[-1])], and here, we discuss the metabolism of the 10B-BPA analog (18F-10B-FBPA). METHODS: Thirty-three cases of primary gliomas were studied by dynamic PET using DL-18F-10B-FBPA or L-18F-10B-FBPA. Dynamic PET images of 18F-10B-FBPA incorporation into tumors were obtained, and the arterial blood samplings were performed in all cases. RESULTS: When the dynamic PET data were represented as Gjedde-Patlak plots, there was a positive slope, suggesting the involvement of the putative metabolic pool of this tracer. A three-compartment model using rate constants (K1, k2 and k3) was used for the kinetic analysis. The accumulation of 18F-10B-FBPA was found to correlate with the degree of malignancy, and the L form of 18F-10B-FBPA was taken up better than was the DL form. The results of dynamic PET analysis suggested that K1 (measuring amino acid transport process) is a major factor determining the accumulation of 18F-10B-FBPA. A comparison of the rate constants revealed that k3 (metabolic process) did not correlate with the degree of malignancy. The absence of evident differences in k3 between DL and L forms suggests that k3 represents phenomena that are not dependent on the native form of L. CONCLUSION: These PET data will be of practical use for diagnosis of malignancy and direct prediction of the effectiveness of boron neutron capture therapy using 10B-BPA.

Astrocytoma↗

[Functional deterioration in adults with cerebral palsy].

We studied functional deterioration in adults with cerebral palsy working at community workshops throughout Japan by interviews and direct examinations. The incidence of functional deterioration was 36%, being higher among those with involuntary movements of the head and neck, abnormal movement patterns when moving and those with reported in poor work environment. In more than one-half case, deterioration was associated with inadequate in noise control, lighting, air circulation from the cooling and heating systems, height of the chairs and desks, and height of VDT screens. Though the factors intrinsic to cerebral palsy were often responsible for the functional deterioration, environmental factors were also significant. Studies on the work environment as well as medical evaluations are needed.

Activities of Daily Living↗

[Increase in pulmonary vascular permeability caused by increased adhesiveness of polymorphonuclear leukocytes and superoxide].

We report that mechanical stimulation of human neutrophils results in their accumulation in isolated rat lungs and in an increase in pulmonary vascular permeability. To determine whether reactive oxygen species were involved in this increase and, if so, whether it is mediate by xanthine oxidase metabolites, we assessed the effect of stimulated and unstimulated neutrophils, and of a superoxide scavenger, superoxide dismutase (SOD), and a xanthine oxidase inhibitor, allopurinol (ALLO) on pulmonary vascular permeability in isolated perfused lungs from Sprague-Dawley rats. Pulmonary vascular permeability in isolated rat lungs was assessed using a filtration coefficient determined by gravimetry. To quantify neutrophil accumulation in the lung, we measured myeloperoxydase (MPO). Neutrophils were stimulated by gentle agitation in a glass container for 10 s and Mac-1 was subsequently upregulated on the surface of the neutrophils. In lungs that received stimulated neutrophils, the pulmonary vascular filtration coefficient was about 5 times higher than in lungs that received unstimulated neutrophils. An increase in filtration coefficient was almost completely blocked by pretreatment with SOD or ALLO. However, the accumulation of stimulated neutrophils was not, or only partly, blocked by SOD or ALLO, respectively. We conclude that the increase in pulmonary vascular permeability caused by mechanically stimulated neutrophils was partly mediated by reactive oxygen species generated via the xanthine oxidase system.

Animals↗

Positron emission tomography-based boron neutron capture therapy using boronophenylalanine for high-grade gliomas: part I.

Determination of tumor boron-10 (10B) levels is required for accurate neutron dosimetry during boron neutron capture therapy. We assessed a new method for quantitative measurement of boronated drug uptake in high-grade gliomas. This method uses positron emission tomography (PET) with fluorine-18-labeled L-fluoroborono-phenylalanine (L-18F-10B-FBPA), which was synthesized as an analogue of L-boronophenylalanine. We studied the accumulation of L-18F-10B-FBPA by PET in patients with high-grade gliomas. Dynamic PET studies of brain tumors revealed that L-18F-10B-FBPA accumulated gradually after bolus injection, and the value of PET activity divided by the integrated plasma activity reached a constant level 42 min after injection, which was defined as the incorporation constant (Ic*). This constant reflected the appropriate L-18F-10B-FBPA accumulation in tumor tissue. Based on the Ic* constant, the methods for estimating tumor 10B concentration were devised. With this method, the estimated values of 10B concentration in gliomas were very close to the 10B levels in surgical specimens. This method was based solely on PET and can potentially provide data that would assist in the selection of patients for future treatment with boron neutron capture therapy after surgical resection of their brain tumors.

Boron↗

Positron emission tomography-based boron neutron capture therapy using boronophenylalanine for high-grade gliomas: part II.

Based on pharmacokinetic findings of fluorine-18-labeled L-fluoroboronophenylalanine by positron emission tomography (PET), methods for estimating tumor 10B concentration were devised. In clinical practice of boron neutron capture therapy (BNCT) for high-grade gliomas, a large amount of L-boronophenylalanine (L-10B-BPA)-fructose solution is used. Under these conditions, a slow i.v. infusion of L-10B-BPA-fructose solution should be performed for BNCT; therefore, the changes over time in 10B concentration in the target tissue were estimated by convoluting the actual time course of changes in plasma 10B concentration with a PET-based weight function including the proper rate constants [K1 (ml/g/min), k2 (min(-1)), k3 (min(-1)), and k4 (min(-1))]. With this method, the estimated values of 10B concentration in gliomas were very close to the 10B levels in surgical specimens. This demonstrated the similarity in pharmacokinetics between fluorine-18-labeled L-fluoroboronophenylalanine and L-10B-BPA. This method, using the appropriate rate constant, permits the determination of tumor 10B concentration and is widely suitable for clinical BNCT, because the averaged PET data are enough to use in future patients without individual PET study.

Boron↗

Evaluation of intraaneurysmal blood velocity by time-density curve analysis and digital subtraction angiography.

PURPOSE: Our purpose was to evaluate intraaneurysmal blood velocity by using time-density curve analysis and digital subtraction angiography. METHODS: In 31 aneurysms, aneurysmal blood velocity was examined with digital subtraction angiography to determine mean transit time (MTF), peak density time (time to peak opacification) (PDT), and time to half-peak opacification (T1/2). Thirty frames per second were acquired, and the time-density curve was calculated. Regions of interest were drawn on the proximal parent artery, on the distal parent artery, and on the aneurysm itself. RESULTS: There was no significant difference in MTT of blood velocity in the proximal site on the parent artery, in the distal site on the parent artery, and in the aneurysm. Similarly, there was no significant difference in PDT in the parent artery, in the distal site on the parent artery, and in the aneurysm; nor was there a significant difference in T1/2 in the parent artery, in the distal site on the parent artery, and in the aneurysm; that is, intraaneurysmal blood velocity was similar to that in the parent artery. PDT and T1/2 of small aneurysms were faster than that of large aneurysms; that is, blood velocity of small aneurysms was faster than that of large aneurysms. CONCLUSION: Intraaneurysmal blood velocity in small aneurysms is similar to that in the parent artery; consequently, the central stream probably reaches the aneurysmal wall. Intraaneurysmal blood velocity in large aneurysms appears to be somewhat slower than that in small aneurysms.

Acute Disease↗

[Assessment of the quality of life (QOL) of prostatic cancer patients under LH-RH analogue treatment].

Assessments of QOL on 53 prostatic cancer patients under LH-RH analogue treatment were evaluated using our own questionnaire. Three months after treatment, the improvement of ADL, appetite, physical symptoms, mental symptoms, sexual life and social life occurred in 24.3%, 51.5%, 44.7%, 29.7%, 7.5% and 35.4% of the patients, respectively. Twelve months after treatment, the improvement of them occurred in 30.9%, 48.4%, 31.7%, 33.7%, 8.3% and 34.4%, respectively. The quality of sexual life for the prostatic cancer patients treated with LH-RH analogue was worse than that of others. LH-RH analogue is effective on QOL in the treatment of prostatic cancer patients.

Antineoplastic Agents, Hormonal↗

[A case of traumatic pseudoaneurysm of the middle meningeal artery treated with endovascular surgery].

A case of traumatic pseudoaneurysm treated by endovascular surgery is reported. The patient was a 20-year-old female. Acute epidural hematoma due to a traffic accident was removed. She was transferred to our hospital in the chronic stage after the operation, because traumatic pseudoaneurysm of the middle meningeal artery had appeared in the same location and had increased in size. We were able to eliminate the aneurysm by endovascular surgical treatment without craniotomy. Endovascular surgical treatment with embolization was less invasive to this patient with traumatic pseudoaneurysm of the middle meningeal artery than the usual surgical procedure.

Accidents, Traffic↗

[Diagnostic usefulness of magnetic resonance cholangiopancreatography (MRCP) in comparison with retrograde cholangiopancreatography (ERCP) for cholelithiasis].

PURPOSE: To elucidate MRCP diagnostic usefulness compared to ERCP. PATIENTS, METHODS: 29 MRCP performed patients diagnosed by ERCP were studied. RESULTS: MRCP showed 92%, 67% sensitivity and 73%, 100% specificity in choledocholithiasis (CBD) and cholecystolithiasis (GB), respectively. MRCP revealed 4.0 false positive and 1.5 false negative in CBD and GB, respectively. MRCP could detect 22 stones out of 33 in CBD and 13 out of 42 in GB. As for the maximal diameter, MRCP depicted 31% for less than 5 mm and 100% for over 6 mm in CBD. In GB, 7% for less than 5 mm, 83% for 6-10 mm and 100% for over 11 mm. CONCLUSION: Although the depection of tiny stone is limited, MRCP is a useful diagnostic tool for cholelithiasis, especially choledocholithiasis.

Adult↗

Anteroposterior gradient of epithelial transformation during amphibian intestinal remodeling: immunohistochemical detection of intestinal fatty acid-binding protein.

To determine whether the remodeling of the well-organized intestinal epithelium during amphibian metamorphosis is regionally regulated along the anteroposterior axis of the intestine, we raised a polyclonal antibody against the Xenopus laevis intestinal fatty acid-binding protein (IFABP), which is known to be specifically expressed in intestinal absorptive cells, and examined immunohistochemically the differentiation, proliferation, and apoptosis of the epithelial cells throughout X. laevis small intestine. During pre- and prometamorphosis, IFABP-immunoreactive (ir) epithelial cells were localized only in the anterior half of the larval intestine. At the beginning of metamorphic climax, apoptotic cells detected by nick end-labeling (TUNEL) suddenly increased in number in the entire larval epithelium, concurrently with the appearance of adult epithelial primordia. Subsequently, the adult primordia in the anterior part of the intestine developed more rapidly by active cell proliferation than those in the posterior part, and replaced the larval epithelial cells earlier than those in the posterior part. IFABP-ir cells in the adult epithelium were first detectable at the tips of newly formed folds in the proximal part of the intestine. Thereafter, IFABP expression gradually progressed both in the anteroposterior direction and in the crest-trough direction of the folds. These results suggest that developmental processes of the adult epithelium in the X. laevis intestine are regionally regulated along the anteroposterior axis of the intestine, which is maintained throughout metamorphosis, and along the trough-crest axis of the epithelial folds, which is newly established during metamorphosis. Furthermore, the regional differences in IFABP expression along the anteroposterior axis of the intestine were reproduced in organ cultures in vitro. In addition, IFABP expression was first down-regulated and then reactivated in vitro when the anterior part, but not the posterior part, of the larval intestine was treated with thyroid hormone (TH) for extended periods. Therefore, it seems that, in addition to TH, an endogenous factor(s) localized in the intestine itself with an anteroposterior gradient participates in the development of the adult epithelium during amphibian metamorphosis.

Animals↗

Effect of estrogen on nitric oxide-induced relaxation of the rabbit urethra.

We evaluated the effect of estrogen on nitric oxide (NO)-mediated urethral relaxation in rabbits. Female New Zealand white rabbits, 4-5 weeks old, were treated with 5 mg/kg estradiol dipropionate (estrogen group) or saline (control group) injected intramuscularly weekly for 2 weeks. Electrical field stimulation (supramaximum voltage, 2 ms pulse duration, 0.3-15 Hz and 3 s train) caused frequency-dependent relaxation of urethral strips in both groups, which was inhibited by Nomega-nitro-L-arginine (L.-NNA). This inhibition was overcome by addition of L-arginine. The relaxation induced by nitrergic nerve stimulation was significantly lower in the estrogen group than in the control group. There was no significant difference in sodium nitroprusside-induced urethral relaxation between the two groups. The production of NO in urethral strips during nitrergic nerve stimulation was evaluated by measuring nitrite/nitrate (NO2-/NO3-) levels in both groups, using microdialysis. The NO2-/NO3- production during electrical field stimulation in the estrogen group was significantly less than that in the control group. The NADPH diaphorase-positive reaction in the control group was greater than that in the estrogen group. The results suggest that estrogen treatment may reduce NO synthase activity, and inhibit the relaxation induced by nitrergic nerve stimulation in rabbit urethral smooth muscle.

Adrenergic beta-Antagonists↗

T cell clonality in synovial fluid of a patient with rheumatoid arthritis: persistent but fluctuant oligoclonal T cell expansions.

Recently, oligoclonal T cell accumulation has been reported in affected joints of patients with rheumatoid arthritis. To characterize such clonally accumulating T cells, we quantitatively monitored their frequency by analyzing TCR B chains. As a result, despite a similar TCR BV usage between synovial fluid (SF) and PBL, obviously skewed TCR BJ usage was detected in SF. Subsequent DNA sequencing demonstrated that the skewed BJ gene usage in SF resulted from clonal T cell accumulation. The complementarity-determining region 3 of TCR B chains of the detected clones appeared to have homologous amino acid sequences. Further, one of the predominant TCR B chains of the clones was identical with that reported previously. In the follow-up study, most of the accumulated clones persisted; however, in some, proportions were drastically decreased or increased during the time course. In particular, one of the clones expanded sixfold, from 11 to 67%, in the BV8-BJ2S3 TCR population of SF, even though the clone was not detected in PBL. In summary, oligoclonal T cell accumulation in SF was persistent, but it appeared to fluctuate independently of the clonality in PBL. The expansion of the clones in SF may occur by antigenic stimuli within the joint.

Amino Acid Sequence↗

Inhibitory effects of HepG2 cell-derived apolipoprotein A-I-containing lipoproteins on cholesteryl ester accumulation in macrophages.

We investigated the mechanisms of inhibitory effects on foam cell formation of apolipoprotein A-I-containing lipoproteins secreted by HepG2 cells (HepG2-HDL) using mouse peritoneal macrophages. When macrophages were incubated with acetylated low-density lipoprotein (acetyl-LDL) in the presence of HepG2-HDL, cholesterol ester (CE) accumulation in cells was reduced by 63%. This inhibitory capacity was almost similar to that of plasma high-density lipoprotein (HDL). When macrophages were converted to foam cells with acetyl-LDL and then reacted with HepG2-HDL or plasma HDL, the HDL-induced CE reduction was 2.2-fold greater than HepG2-HDL. Similar results were obtained using apo E-free HepG2-HDL. Since the inhibitory effect of HDL on acetyl-LDL-induced CE accumulation in macrophages is due largely to its cholesterol efflux capacity, these results suggest the presence of an additional mechanism for the inhibition of CE accumulation by HepG2-HDL. To investigate the mechanism, acetyl-LDL was reisolated from HepG2-HDL by Sephacryl S-300 gel filtration after incubation in a cell-free system. Reisolated acetyl-LDL showed a significant reduction in electrophoretic mobility. The extent of CE accumulation by reisolated acetyl-LDL was reduced by 20% compared with control acetyl-LDL. Moreover, its endocytic degradation by macrophages was reduced by 28%. HepG2-HDL also inhibited macrophage degradation of acetyl-LDL as well as oxidized LDL, a likely atherogenic lipoprotein. This inhibitory effect was ascribed to the HepG2-HDL subfraction containing pre-beta HDL. Our results indicated that apo A-I-containing lipoproteins as a physiological model of nascent HDL may inhibit foam cell formation by reducing ligand activity of atherogenic lipoproteins. These data possibly suggest inhibitory function of nascent HDL for the formation of foam cells in vivo.

Animals↗