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Biomedical subjects

K Fujii

Publications and source records attributed to K Fujii.

At least 523 records · Page 29Linked to original sources

Application of the ILISPOT-IDIP system for the enumeration of different sizes of IgA spot forming cells in the murine small and large intestine.

The immunofluorescence-linked immunospot (ILISPOT) assay associated with the immunofluorescence digital image processing (IDIP) system was originally developed in our laboratory to allow enumeration of immunoglobulin (Ig) producing, spot forming cells (SFC) in a more objective and quantitative manner. In this study, the ILISPOT-IDIP system was further advanced in order to analyze different sizes of SFC (e.g., IgA producing cells) including large (L), medium (M), and small (S) cells which correspond to high (> 2.4 pg), medium (1.2-2.4 pg) and low (< 1.2 pg) IgA secreting cells by the adaptation of real time image processor and intensified video camera system. When the ILISPOT-IDIP system was used to characterize the frequency of IgA secreting cells among mononuclear cells isolated from different parts of the murine gastrointestinal (GI) tract including the small (upper, middle and lower sections) and large (colon and rectum) intestine, the small intestine contained higher numbers of IgA SFC (approximately 8.4 x 10(5) SFC/10(6) cells) when compared with large intestine (approximately 1.3 x 10(5) SFC/10(6) cells). Among the 3 areas of small intestine, the upper (approximately 3.7 x 10(5) SFC/10(6) cells) and middle (approximately 2.4 x 10(5) SFC/10(6) cells) parts had higher numbers of IgA SFC when compared to the lower small (approximately 2.3 x 10(5) SFC/10(6) cells) intestine. When these IgA producing cells in different parts of the intestine were classified into three groups according to the size of individual spots, the upper and middle intestine contained higher frequencies of large (approximately 20%) and medium (approximately 20%) SFC which corresponded to high and medium IgA secretors in comparison to the lower small (approximately 9%) and large (approximately 6%) intestine. In contrast, the lower small and large intestine were dominated by small SFC since approximately 85% of IgA producing cells were categorized as low secretors. Using the advanced ILISPOT-IDIP system, a unique distribution of different sizes (or secretion rates) of IgA producing SFC was elucidated in the different regions of the mouse small and large intestine.

Animals↗

Modification of integrin-mediated cell attachment to substrata by serine proteinases in the presence and absence of divalent cations.

The sensitivity to serine proteinases of cellular proteins involved in cell-matrix adhesion was investigated using C32 melanoma cells. Cells dissociated from monolayers by the metal chelator ethylenediaminetetraacetic acid were incubated with proteolytic enzymes, and then attachment was quantified by standard cell adhesion assays. The effect of proteinases was found to depend on the presence of Ca2+ in the incubations. Incubation with 100 nM trypsin or chymotrypsin for 1-2 h (37 degrees C) in the absence of Ca2+ reduced cell attachment to vitronectin (Vn), fibrinogen (Fb), laminin, and fibronectin by approximately 80, 80, 40, and 30%, respectively. Viability studies indicated that such treatment with proteinases was not cytotoxic. Inclusion of 0.1 nM CaCl2 in the incubations prevented the loss in attachment to all substrata. In the case of Fb, proteinase treatment in the presence of Ca2+ had an additional effect; it improved cell attachment to this substratum by about 50%. C32 cells have been shown to express the integrin alpha v beta 3 (Vn receptor) which mediates attachment to Vn and Fb in a GRGDS-sensitive manner. Attachment of C32 cells to Vn and Fb prior to proteinase treatment and after proteinase treatment in the presence of Ca2+ was 90% inhibited by the addition of GRGDS peptide to the attachment assays. These results suggest that the adhesion observed both before and after proteinase treatment was mediated by this integrin. Analysis of the Vn receptor from proteinase-treated cells by immunoblotting of cell extracts and by SDS gel electrophoresis of immunoprecipitated receptor revealed no detectable change in either the alpha v or beta 3 subunit that correlated with loss in attachment. Similarly proteinase treatment in the presence of Ca2+ did not produce detectable alterations in the subunits which might correlate with the improved attachment to Fb. Consistent with these results, an enzyme-linked immunoassay to quantify cell surface receptors revealed little difference in the amount of Vn receptor on cells treated with proteinase in the presence or absence of Ca2+. Degradation of the alpha v subunit was demonstrated, however, at proteinase concentrations higher than those required to affect cell attachment. Thus, treatment of cells with serine proteinases can affect integrin-mediated attachment to matrix proteins in a manner moderated by Ca2+, but the alterations in attachment do not appear to be accompanied by detectable proteolytic modification of the integrin.

Amino Acid Sequence↗

Reconstruction of the posterior cruciate ligament with LAD-augmented semitendinosus and gracilis tendons: a preliminary report.

We present our technique for reconstruction of the posterior cruciate ligament (PCL) using the semitendinosus and gracilis tendons with the Kennedy ligament augmentation device (LAD). The safe and excellent exposure of the posterior aspect of the knee allowed us to identify the most isometric position in the intercondylar notch of the femur. In addition to this advantage, firm fixation of the LAD-augmented tendons with staples prevented the tibia from sagging posteriorly during early protected motion of the knee. Evaluation of 12 patients followed for more than 2 years showed 9 (75%) good results. In this small series no correlation was found between clinical results and the number of major structures injured, indicating that postoperative care is as important as isometric placement of the PCL in obtaining satisfactory results.

Adolescent↗

Repeat angiography and magnetic resonance imaging (MRI) of dissecting aneurysms of the intracranial vertebral artery. Report of four cases.

We here present 4 cases with dissecting aneurysm (DA) of the intracranial vertebral artery, who were followed up by repeat cerebral angiography and MRI. The patients consisted of 2 males and 2 females, and the mean age was 43 years. Two cases were associated with polyarteritis nodosa (PN) and hypertension, respectively. Three of the cases developed subarachnoid haemorrhage (SAH), while the other one suffered from lateral medullary syndrome. In cerebral angiography, "pearl and string" signs were revealed in all cases, while a "double lumen" indicating a true diagnostic sign of DA was demonstrated in only one case. Repeat angiography showed that a bleb formation with a bulging of the aneurysmal sac was seen in 2 cases, and an irregularity of the wall in one case. On the other hand in one case, the ectatic part shrank, while the stenotic part was restored. In magnetic resonance imaging (MRI), a hyperintensity mass on T 1-weighted image (T 1-WI) adjacent to flow void suggesting either an intramural haematoma or a linear shape hyperintensity on T 1-WI were demonstrated in 3 cases. In the follow up MRI done in 2 cases, a serial change in the intensity from iso-intensity to hyperintensity on T 1-WI was observed in one case suggesting intramural haemorrhage, while an enlargement of the ectasic flow void was seen in the other case. Three of 4 cases were operated on by trapping of the aneurysms. One, who had systemic vascular diseases due to PN, and repeat angiography showed a regression of the aneurysm, was conservatively treated.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

De novo aneurysm formation and aneurysm growth following therapeutic carotid occlusion for intracranial internal carotid artery (ICA) aneurysms.

We herein report the long term results of 27 intracranial internal carotid artery (ICA) aneurysms treated by indirect surgery such as a trapping of the aneurysm or carotid ligation either with or without EC-IC bypass. These patients were then followed for a mean period of 10 years. Seventy-four percent of the aneurysms were categorized as either being giant or large. Late complications were observed in 7 patients. A rupture of newly formed aneurysms at the anterior communicating artery occurred in 2 cases 8 or 9 years after either trapping or performing a ICA ligation, respectively. In these patients, previous angiography could not reveal any abnormalities at the anterior communicating artery. A rupture of a contralateral ICA aneurysm was seen in a patient whose ipsilateral ICA was ligated for a ICA aneurysm 22 years previously. An enlargement of the contralateral giant cavernous ICA aneurysm became symptomatic 6 years after a partial ligation of the ICA combined with an EC-IC bypass for a giant cavernous carotid artery aneurysm on the other side. Two cases of sudden death occurred in a young patient and an elderly patient with a small anterior communicating artery aneurysm, 9 and 19 years respectively, after trapping of the ICA aneurysms, although the cause could not be definitely ascertained. Rebleeding occurred in one patient who died 8 years after a carotid ligation and a partial clipping of the ICA aneurysm. Haemodynamic stress may therefore play a major role in inducing new aneurysms or growing aneurysms.(ABSTRACT TRUNCATED AT 250 WORDS)

Carotid Artery Diseases↗

Bone-grafting for acetabular deficiency during primary and revision total hip arthroplasty. A radiographic and clinical analysis.

The use of bone-graft to augment the deficient acetabulum in primary and revision total hip arthroplasty (THA) is controversial. To identify factors affecting cup loosening in patients who received a bone-graft during THA, two orthopaedic surgeons retrospectively examined sequential radiographs. The surgeons also obtained independent computer measurements of hip center and cup abduction migration from preoperative, initial, and latest postoperative radiographs. Variables studied included host factors, graft factors, and technique factors. All conclusions were based on Kaplan-Meier log-rank analysis to account for differing lengths of follow-up periods among the cases. The authors report a series of 74 consecutive cases with a minimum 24-month follow-up period (mean, 40 months). All grafts appeared to unite. The clinicians found 80% stable cups, 8% possibly loose cups, and 12% (n = 9) definitely loose cups. In retrospect, technical errors were seen in six loose cups. Five revisions for loosening (6.7% of cases) were performed. Computer measurement found cup loosening in a higher percentage of cases than detected by the clinicians and did so an average of 18 months sooner. Acetabular cup loosening was associated with the American Academy of Orthopaedic Surgeons type III defects, use of allograft versus autograft, and initial cup abduction of 50 degrees or more. Kaplan-Meier survivorship analysis found 31% of cups radiographically loose and 15% revised at 5 years or more since surgery. Acetabular bone-grafting is technically demanding and should be employed when alternative reconstructions will not give a durable result.

Acetabulum↗

A rapid molecular diagnosis of posttransfusion graft-versus-host disease by polymerase chain reaction.

A woman with recurrent Paget's disease of the vulva developed acute graft-versus-host disease (GVHD) 12 days after radical surgery and massive blood transfusion. Molecular diagnosis of lymphocyte chimerism in the peripheral blood was made by polymerase chain reaction (PCR) directed against a Y chromosome-specific sex-determining region Y (SRY) gene. PCR with skin biopsy after onset of GVHD also revealed infiltration of SRY-positive donor lymphocytes. The diagnosis was confirmed by HLA-DNA typing with PCR-sequence-specific oligonucleotide that revealed the presence of complex HLA-DR chimerism in the peripheral lymphocytes collected after onset of GVHD. The use of SRY-directed PCR is a rapid technique for the early diagnosis of acute posttransfusion GVHD in female patients.

Base Sequence↗

Urinary excretion of hexafluoroisopropanol glucuronide and fluoride in patients after sevoflurane anaesthesia.

The excretion of sevoflurane metabolites in the urine collected every 12 h after sevoflurane anaesthesia was measured by ion exchange chromatography. A metabolite, which was converted on incubation with glucuronidase to hexafluoroisopropanol was detected in the urine. The maximum excretion was found in the first 12 h after anaesthesia, none was found in the last collection 3 days after anaesthesia. The excretion half-life for the metabolite was calculated to be 55 h. A significant increase in the urinary excretion of organic and inorganic fluoride was also observed during the first 12 h after anaesthesia. The cumulative organic and inorganic fluoride excretion in the 3 days after sevoflurane anaesthesia was 1588 and 856 mumol, respectively (ratio = 1.85). The excreted half-lives for organic and inorganic fluoride were calculated to be 4028 and 2069 min, respectively. Our study showed that a hexafluoroisopropanol glucuronide is excreted in the urine, and the major part of urinary metabolites of sevoflurane, organic and inorganic fluoride, are excreted within 2 days of sevoflurane inhalation in man.

1-Propanol↗

In vitro antimicrobial activity of benzoxazinorifamycin, KRM-1648, against Mycobacterium avium complex, determined by the radiometric method.

MICs of a newly developed benzoxazinorifamycin derivative, KRM-1648, for Mycobacterium avium complex (MAC) were determined by the BACTEC 460 TB system and compared with those of other known antimicrobial agents. The radiometric method gave a fast, accurate, and reproducible MIC for each antimicrobial agent. MICs of KRM-1648 for 30 strains of MAC (10 strains each of M. avium isolated from AIDS and non-AIDS patients and of Mycobacterium intracellulare isolated from non-AIDS patients) were measured. The MICs, ranging from 0.004 to 0.0625 microgram/ml, were the lowest of all tested drugs, including rifampin, rifabutin, streptomycin, kanamycin, isoniazid, ethambutol, ofloxacin, ciprofloxacin, sparfloxacin, and clarithromycin. The MICs were 2 to 512 and 1 to 32 times lower than those of rifampin and rifabutin, respectively. With rifampin and ethambutol, there were some differences between the MICs for M. avium isolated from AIDS patients (American) and those for M. avium from non-AIDS patients (Japanese). Moreover, appreciable differences between the MICs of some drugs against M. avium and M. intracellulare isolated from non-AIDS patients were found. Many strains of M. avium were more susceptible to ofloxacin than M. intracellulare, but, conversely, M. avium was more resistant to rifampin, streptomycin, ethambutol, and clarithromycin than M. intracellulare.

Acquired Immunodeficiency Syndrome↗

Age-related changes in endothelium-dependent hyperpolarization in the rat mesenteric artery.

This study was designed to determine the age-related changes in the endothelium-dependent hyperpolarization to acetylcholine (ACh) and its contribution to relaxation in the isolated mesenteric artery from normotensive and hypertensive rats. Membrane potentials and contractions were recorded in arteries from male Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR) that were 5-6 wk old (young), 6-8 mo old (adult), and 20-26 mo old (aged). Endothelium-dependent hyperpolarizations produced by ACh, applied both at the resting state of the membrane and under conditions of depolarization with norepinephrine (10(-5) M), were markedly impaired in aged WKY rats, adult SHR, and aged SHR. Endothelium-dependent relaxations to ACh in arterial rings precontracted with 10(-5) M norepinephrine were also impaired in aged WKY rats, adult SHR, and aged SHR even in the presence of indomethacin. Furthermore, in these rats, N omega-nitro-L-arginine, an inhibitor of nitric oxide formation, showed potent inhibitory effects on the relaxations, whereas the 20 mM high K+ solution that reduces hyperpolarization had less pronounced effects. Hyperpolarizations and relaxations to cromakalim (10(-5) M), a K(+)-channel opener, were on the whole preserved in aged rats. It would thus appear that the endothelium-dependent hyperpolarization to ACh is reduced with aging as well as by hypertension, and this would, in part, account for the impaired relaxation to ACh in arteries of both aged rats and hypertensive rats.

Acetylcholine↗

Time course of functional improvement in stunned myocardium in risk area in patients with reperfused anterior infarction.

BACKGROUND: The beneficial effect of coronary reflow on myocardial salvage may be assessed more accurately than in previous studies if the size of risk area is taken into account, particularly because the size of risk area varies significantly among patients. In this study, the risk area was determined with myocardial contrast echocardiography to investigate the time course of functional recovery of postischemic myocardium within the risk area in patients with reperfused anterior myocardial infarction. METHODS AND RESULTS: The study population consisted of 21 patients with anterior myocardial infarction who achieved coronary reflow within 6 hours of onset by means of thrombolysis or coronary angioplasty. Myocardial contrast echocardiography was performed with the injection of hand-agitated Haemaccel (5 ml) into the right and left coronary arteries before coronary reflow, and the risk area was defined as the area of contrast perfusion defect in the apical long-axis view. The ratio of the endocardial length of abnormal contraction (dyskinesis/akinesis) segment to that of contrast defect segment (AS/CD) was determined at days 1, 2, 3, 7, 14, and 28 of reflow. Before reflow, the length of contrast defect correlated well with the segment length of dyskinesis/akinesis. The values for AS/CD in patients with successful reperfusion significantly and progressively decreased until day 14; 1.00 +/- 0.02 at day 1, 0.93 +/- 0.11 at day 2 (p < 0.05 versus day 1), 0.84 +/- 0.16 at day 3 (p < 0.05 versus day 2), 0.80 +/- 0.13 at day 7 (p < 0.01 versus day 2), 0.73 +/- 0.10 at day 14, and 0.72 +/- 0.10 at day 28. Greater improvement in function was obtained in patients reperfused within 4 hours than in those reperfused at > or = 4 hours (AS/CD at day 28, 0.64 +/- 0.12 versus 0.75 +/- 0.09, p < 0.05). CONCLUSIONS: Thus, a significant amount of myocardium, an average of 28% in segment length of the risk area, is salvaged in patients with reperfused anterior myocardial infarction. Major functional improvement seems to be achieved within 14 days of reflow.

Adult↗

Voltage-dependent Ca2+ channels in resistance arteries from spontaneously hypertensive rats.

Alterations in voltage-dependent Ca2+ channels in the arterial smooth muscle cells of spontaneously hypertensive rats (SHR) were investigated using the whole-cell voltage clamp and compared with Wistar-Kyoto (WKY) rats. Single cells were freshly isolated from resistance mesenteric arteries from 4- to 5-week-old (young) and 16- to 18-week-old (adult) SHR. Elevated blood pressure was only evident in adult SHR, not in young SHR. In young rats, the Ca2+ channel current density (current amplitude normalized by cell capacitance) was significantly higher (P < .01) in SHR than in WKY rats at the command potential of -10 mV or higher (with 50 mmol/L Ba2+): The current density at 20 mV was -16.8 +/- 1.1 pA/pF in SHR (n = 38 cells) and -11.0 +/- 0.8 pA/pF in WKY rats (n = 30 cells). In adult rats, the difference in current densities disappeared: -15.9 +/- 1.3 pA/pF in SHR (n = 25 cells) and -15.6 +/- 1.5 pA/pF in WKY rats (n = 29 cells). The ratio of maximal amplitude of T-type current to that of L-type current was low in young SHR (0.10 +/- 0.01) compared with the other three groups (0.16 to 0.20). Neither the activation curve nor the steady-state inactivation curve of SHR was different from that of age-matched WKY rats. However, the activation curves in adult rats were shifted to a hyperpolarized direction compared with those of young rats in both strains. These results suggest that the increased activity of voltage-dependent L-type Ca2+ channels of resistance arteries in young SHR may be related to the development of hypertension. The changes observed in adult rats may be due to a secondary modification of the channel during maturation and the presence of hypertension.

Animals↗

Angiotensin converting enzyme inhibitors attenuate ischemic brain metabolism in hypertensive rats.

BACKGROUND AND PURPOSE: Angiotensin converting enzyme (ACE) inhibitors are expected to modulate neuronal activities. The present study was designed to examine the beneficial effects of ACE inhibitors on microcirculation and metabolism in the ischemic brain. METHODS: Cerebral ischemia was developed for 60 minutes in spontaneously hypertensive rats (SHR, n = 35) by bilateral carotid artery occlusion. ACE inhibitor (0.1 or 10 mg/kg SQ 29,852 or captopril) were intravenously injected 15 minutes before cerebral ischemia. Cerebral blood flow to the parietal cortex was measured with the H2 clearance technique. Lactate, pyruvate, and ATP in the brain were estimated by the enzymatic method. RESULTS: Before cerebral ischemia, high doses of both SQ 29,852 and captopril significantly decreased mean arterial pressure by 15 to 25 mm Hg and reduced cerebral vascular resistance by 13% to 17% of the resting values. Cerebral blood flow and arterial pressure during ischemia were not altered by these ACE inhibitors. After 60 minutes of cerebral ischemia, tissue lactate in vehicle-treated SHR increased 6.6-fold and ATP decreased to 65% of the control values. Administration of SQ 29,852 or captopril significantly reduced the lactate levels to 1.6- to 3.1-fold and well preserved the ATP levels to 82% to 93% of the control. CONCLUSIONS: These results suggest that inhibition of ACE activities may be protective for cerebral metabolism against ischemic insult.

Adenosine Triphosphate↗