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Biomedical subjects

Y Fukushi

Publications and source records attributed to Y Fukushi.

At least 37 records · Page 2Linked to original sources

Galactosylgloboside expression in seminoma. Inverse correlation with metastatic potential.

BACKGROUND: Altered glycosylation is a common phenotype expressed in essentially all types of human cancer and has been found to be correlated closely with the invasive and metastatic properties of a given tumor. Because there was no prognostic information concerning aberrant glycosylation of seminoma, the authors studied this topic. METHODS: Glycosphingolipid (GSL) composition of orchiectomy samples of seminoma were analyzed systematically. GSL patterns from seminoma samples of the following three groups were compared after a 44-month postoperative period: Stage I disease with no evidence of metastasis during the 44-month postoperative period, Stage I with metastatic relapse during this period, and Stage II with retroperitoneal lymph node metastasis. Unknown GSLs detected were analyzed chemically by 1H-nuclear magnetic resonance spectroscopy and mass spectrometry. RESULTS: All nonmetastatic seminomas (n = 12) contained a GSL band that was identified as galactosylgloboside (Gb5; Gal beta 1-->3GalNAc beta 1-->3Gal alpha 1-->4 Gal beta 1-->4Glc beta 1-->1Cer). All metastatic seminomas (n = 5) lacked this GSL, although the sample sizes were admittedly small. CONCLUSION: Only the presence or absence of galactosylgloboside (Gb5), but of no other GSL or gangliosides, clearly correlated with metastatic potential in patients with seminoma. This observation is useful in the estimation of prognosis of patients with seminoma, especially those with Stage I disease.

Adult↗

Antimicrobial stress compounds from Hypochoeris radicata.

The exudates of Hypochoeris radicata leaves stressed with cupric chloride afforded two eudesmane- and guaiane-type sesquiterpenes, and two alkenals, (2E,4E)-6-hydroxyhexadienal and (2E,4E)-hexadienedial (mucondialdehyde). Their structures were determined by spectroscopic and synthetic methods.

Antifungal Agents↗

Ca2+ entry through the store-mediated pathway directly activates only the K+ current but the subsequent Ca2+ release from the store activates both K+ and Cl- currents in submandibular gland acinar cells of the rat.

The store-mediated Ca2+ entry was detected in single and cluster of rat submandibular acinar cells by measuring the Ca2+ activated ionic membrane currents. In the cells where intracellular Ca2+ was partly depleted by stimulation with submaximal concentration of acetylcholine (ACh) under a Ca2(+)-free extracellular condition, an employment of external Ca2+ in the absence of ACh caused a sustained increase of the K+ current without affecting the Cl- current. A renewed ACh challenge without external Ca2+ caused repetitive spikes of both K+ and Cl- currents due to the Ca2+ release. SK & F 96365 inhibited the generation of the sustained K+ current and refilling of the Ca2+ store following the Ca2+ readmission. It is suggested that the Ca2+ enters the cell through the store-mediated pathway new the K+ channels and is taken up by the store. Thus, only Ca2+ released from the store can activate both the K+ and Cl- currents.

Acetylcholine↗

Sr2+ can pass through Ca2+ entry pathway activated by Ca2+ depletion, but can be hardly taken up by the Ca2+ stores in the rat salivary acinar cells.

When Sr2+ was introduced to the external solution after the depletion of Ca2+ from stores of submandibular acinar cells by ACh stimulation, Sr2+ entered cytoplasm of the cell, like the case of Ca2+. SK&F 96365, a Ca2+ channel blocker, or Ni2+ blocked this divalent cation entry. Sr2+ entering the cell continued to increase to a steady level, after the cessation of stimulation, when Sr2+ was present in the external solution, unlike the case of Ca2+. Ca2+ entered cells which had been stimulated with ACh in Sr(2+)-containing external solution. In the cells to which Sr2+ has been applied after the store depletion with ACh, Sr2+ cannot be released by the renewed ACh stimulation, unlike the case of Ca2+. 89Sr2+ uptake by the parotidic microsomal fraction 100 min after addition of ATP was 15.08 +/- 0.70 nmol/mg protein, whereas 45Ca2+ uptake was 144.19 +/- 16.93 nmol/mg protein. It was concluded that in the salivary acinar cells Sr2+ can be a substituent for Ca2+ in the mechanism of entry from the extracellular fluid but cannot be in the mechanism of uptake into the stores.

Animals↗

Stimulated Ca2+ entry activates Cl- currents after releasing Ca2+ from the intracellular store in submandibular gland cells of the rat.

In order to examine whether Ca2+ entry is directly involved in controlling exocrine secretion, the Ca(2+)-activated Cl- currents were recorded in single and clusters of rat submandibular gland cells using the whole-cell patch-clamp method. Extracellularly applied acetylcholine (ACh, 10 nM) as well as intracellularly applied GTP gamma S and InsP3 caused repetitive transients of the Cl- currents activated by intracellular Ca2+. These responses occurred also in the absence of external Ca2+, but disappeared after several minutes. Readmission of Ca2+ to the extracellular solution restored the repetitive current transients, while introduction of Sr2+ failed to restore the current signals in spite of the presence of Sr2+ entry detected by microfluorimetry. On the other hand, direct application of Sr2+ to the cell inside caused activation of the Cl- currents although less effectively than Ca2+. When Ca2+ was introduced to the extracellular solution during an interruption of ACh stimulation after the ACh-induced depletion of intracellular Ca2+ store, the Cl- current was not elicited. However, a subsequent challenge with ACh at the same concentration in the absence of extracellular Ca2+ caused repetitive transient Cl- currents. The results suggest that in this cell type the stimulated Ca2+ entry does not by itself activate the Cl- currents but activates them indirectly by triggering Ca2+ release from the intracellular Ca2+ store which may take up Ca2+ soon after the Ca2+ entry.

Acetylcholine↗

Modulation of ischemia-reperfusion-induced hepatic injury by Kupffer cells.

To elucidate the role of Kupffer cells in ischemia-reperfusion-induced hepatic injury, hepatic injury induced by ischemia-reperfusion was analyzed after modulation of Kupffer cell function. Ischemia of the liver was performed by occlusion of both the portal vein and hepatic artery, which enter into the left lateral and median lobes of the liver. Blood flow in the ischemic lobe was reduced, in contrast to an increased blood flow in the nonischemic lobe during occlusion of the veins. Although hepatocyte damage was not demonstrated by ischemia for < 60 min, hepatic injury was found after reperfusion of the liver, and activation of Kupffer cells was morphologically demonstrated by electron microscopies. Suppression of Kupffer cells, induced by previous administration of gadolinium chloride or latex particles, reduced the grade of hepatic injury induced by ischemia-reperfusion. On the other hand, stimulation of Kupffer cell phagocytosis, induced by administration of latex particles at the time of reperfusion, aggravated the ischemia-reperfusion-induced hepatotoxicity, which was then reduced by simultaneous administration of superoxide dismutase. Kupffer cells, isolated from the rats treated with the ischemia-reperfusion procedure, have been found to release increased amounts of oxygen radical intermediates. These results suggest that hepatic injury induced by ischemia-reperfusion is modulated by the function of Kupffer cells and that superoxide anion released from Kupffer cells could play an important role in ischemia-reperfusion hepatic injury.

Animals↗

Plasma peroxidized low-density lipoprotein with hydroperoxidized cholesteryl linoleates estimated in patients with familial hypercholesterolemia.

A study was conducted to ascertain the presence of peroxidized low-density lipoprotein (LDL) in plasma and to determine the chemical structure of the peroxidized LDL so that the mechanism by which vascular complications develop may be elucidated in patients with familial hypercholesterolemia (FH). Family trees showing hypercholesterolemia and moderate grade thickness of the Achilles' tendon were confirmed in all patients. Cholesteryl ester, triglycerides (TG), free fatty acid (FFA), free cholesterol (FC), and phospholipids in normal LDL were stained on a thin-layer chromatography (TLC) plate, but from peroxidized LDL, spot X1 between TG and FFA, and spot X2 between FFA and FC were identified clearly except for the lipids recognized in normal LDL on the TLC plates. From intermediate-density lipoprotein (IDL), LDL1, and LDL2 of FH patients, spot X1 was clearly recognized. Hydroperoxidized cholesteryl linoleate (HPO-CL) was estimated and identified from Cu(2+)-treated standard CL, Cu(2+)-treated LDL and LDL obtained from FH patients by means of high-pressure liquid chromatography, mass spectrometry, nuclear magnetic resonance, positive reaction of p-methoxydiphenylpyrenyl-phosphine and various chemical reactions. The percentage of HPO-CL to total lipids in LDL was in the order of IDL < LDL1 < LDL2 in FH patients, and it was markedly higher in FH patients when compared to healthy persons. Spot X1 obtained from Cu(2+)-treated CL, from Cu(2+)-treated normal LDL and from LDL of FH patients was chemically identical, consisting of at least 4 kinds of HPO-CL isomer. LDL2-cholesterol levels were higher in FH patients than those of LDL1-cholesterol, whereas in healthy persons LDL1 cholesterol levels were higher compared to those of LDL2 cholesterol. The capacity of LDL2 for peroxidization by Cu2+ was greater than that of LDL1 in healthy persons. Biological functions, such as acceleration of platelet aggregation, great internalization into macrophages and injuries of the arterial endothelia, were reported for LDL with HPO-CL in our previous experiments. Thus, the existence of peroxidized LDL with HPO-CL may be one of the important risks for vascular complications in FH patients.

Adult↗

Malignant pleural mesothelioma presenting as achalasia.

A 65-year-old man with an occupational history of asbestos exposure developed dysphagia and vomiting. Clinical examinations at onset revealed a dilated esophagus with smooth narrowing at the gastroesophageal junction and no apparent tumor in and around the esophagus. Achalasia was suspected. Dysphagia progressed gradually and examinations performed three months after the onset disclosed a tumor in the pleural and the peritoneal cavities. At laparotomy, the tumor extended from the pleural cavity into the peritoneal cavity. Histological examination of the biopsied specimen demonstrated malignant mesothelioma. We report the first case of malignant pleural mesothelioma presenting as achalasia.

Aged↗

[Immunohistochemical study of globotriaosyl ceramide (Gb3) in testicular tumors].

Testicular tumor tissues and various normal tissues were examined by immunohistochemical techniques for expression of globotriaosylceramide (Gb3) using anti-Gb3 monoclonal antibody (1A4). In formalin fixed paraffin-embedded specimens (31 cases), Gb3 was positive for every kind of elements of testicular germ cell tumors, especially it's incidence of positive staining were 100% in seminomas (16/16) and embryonal carcinomas (6/6). On the other hand, four cases of testicular malignant lymphoma were all negative for Gb3, and some normal testicle, liver, kidney, pancreas, spleen, ileum and rectum showed very weak reactivity with 1A4. In frozen materials (10 cases), similar incidence of positive staining with that of paraffin-embedded specimens were observed in drying only manner, 4% parafolm-aldehyde fixation following drying and 10 min. of acetone fixation following drying. Incidence of very strong staining in testicular germ cell tumors were as follows, 7/10 in 4% parafolmaldehyde fixation, 6/10 in drying only, 4/10 in 10 min. of acetone fixation and 7/22 in paraffinembedded specimens. Fixation with 30 min. of acetone fixation and 10 min. of 90% ethanol and treatment with 10 min. of chloroform:methanol (2:1, v/v) extremely decreased the reactivity with 1A4, suggesting that Gb3 exists mainly as glycolipid. Gb3 may thus be a useful histological marker for testicular germ cell tumor, and the tissue should be treated with the process of freezing followed by 4% parafolm-aldehyde fixation, drying only or 10 min. of acetone fixation, and formalin fixed paraffin-embedded specimens were also available enough.

Biomarkers, Tumor↗

Globotriaosyl ceramide glycolipid in seminoma: its clinicopathological importance in differentiation from testicular malignant lymphoma.

Glycolipids were biochemically extracted from 14 specimens of seminoma, 2 of testicular malignant lymphoma (both of which were difficult to differentiate from seminoma with a high mitotic index) and 4 of normal testicle. The pattern of their expression was compared. Marked accumulation of globotriaosyl ceramide was observed in seminoma but it was present in a small amount in testicular malignant lymphoma. Differentiation between seminoma and malignant lymphoma is sometimes difficult by histopathological findings but it is considered to be greatly facilitated by examination of the pattern of glycolipid expression.

Antigens, Differentiation, B-Lymphocyte↗

The pH dependent uptake of enoxacin by rat intestinal brush-border membrane vesicles.

The mechanism of the intestinal transport of enoxacin, an orally active fluoroquinolone antibiotic, has been investigated using brush-border membrane vesicles isolated from rat small intestine. The initial rate and time-course of enoxacin uptake were considerably dependent upon the medium pH (pH 5.5 greater than pH 7.5) and upon the percent ionization of the carboxyl group (pKa 6.2, anionic charge), namely, the degree of uptake of cationic form was higher than that of the zwitterionic form. There was evidence of transport into the intravesicular space as shown by the effect of extravesicular medium osmolarity on enoxacin uptake at steady state (30 min). This transport across the brush-border membrane was stimulated by the valinomycin-induced K(+)-diffusion potential (interior negative) and an outward H(+)-diffusion potential. Furthermore, changing the pH of the medium from 5.5 to 7.5 significantly decreased the effect of valinomycin-induced K(+)-diffusion potential on the enoxacin uptake. These results suggest that the uptake behaviour of the cationic form of enoxacin plays an important role in the intestinal absorption process of enoxacin.

Animals↗

Glycolipid expression in prostatic tissue and analysis of the antigen recognized by antiprostatic monoclonal antibody APG1.

The expression patterns of glycolipid from prostatic hyperplasia, prostatic cancer and normal prostate tissue were observed. A further analysis of antigen recognized by mouse monoclonal antibody APG1, which was gained by immunizing glycolipids extracted from human prostate cancer, was also performed. In cancer tissue, both of the lactosyl and globoside series glycolipids were found to be generally reduced, although in the ganglioside series, GM3 and GD3 were not reduced and only the glycolipids with longer chains than GD2 were found to be reduced. These results indicated that the inhibition of sugar chain elongation, but not sialylation, was the main synthetic change occurring with carcinogenesis of the human prostate. APG1 reacted with only two bands near GM2 and GD2 of the ganglioside fraction on a thin-layer chromatography plate, but it did not react with any of the known gangliosides of the ganglioside series including GM2 and GD2. Histochemically, APG1 showed intense reaction only in frozen tissue sections of human prostate, and the reactivity decreased with the increasing grade of cancer. Therefore, this antigen was considered to be a prostate-specific and differentiated antigen reacting with nonganglioseries gangliosides.

Antibodies, Monoclonal↗

Changes in glycolipids in human renal-cell carcinoma and their clinical significance.

The expression patterns of glycolipids of human renal-cell carcinoma were studied in primary tumors from 23 cases and 5 metastatic lesions from 4 cases using HPTLC (high-performance thin-layer chromatography). The expression pattern of glycolipids in primary tumors was characteristic of the histological cell type. In granular-cell carcinoma, lactosylceramide (CDH), GM3 and the longer-chain gangliosides (gangliosides migrating more slowly than GM3) increased, although in clear-cell carcinoma, CDH and other glycolipids tended to decrease. Globoside decreased in all cases but one, irrespective of cell type. In metastatic lesions of the clear-cell type, the prominent increase in longer-chain gangliosides was characteristic. Furthermore, the same expression pattern as that of metastastatic lesions was shown in 3 of 14 patients with primary clear-cell carcinoma, all of whom developed metastases soon after radical nephrectomy. These studies indicate that the increased expression of the longer-chain gangliosides in primary tumor is one of the factors associated with high metastatic potential, and also predict early post-operative development of metastasis.

Aged↗

Effects of low-density lipoprotein from normal rabbits on hypercholesterolemia and the development of atherosclerosis.

To clarify whether or not normal LDL separated from normal rabbits is atherogenic, 0.75 mg of normal LDL-cholesterol in 6 ml of 0.85% saline and 6 ml of saline alone were injected every day for 5 weeks into the auricular vein of two groups of rabbits, respectively. The inoculated rabbits were fed a standard diet containing 0.5 and 1% cholesterol. Blood was drawn before injection and at 1, 2, 3 and 5 week intervals thereafter. After 5 weeks, all rabbits were sacrificed. Following exsanguination, the aorta was stained by Sudan III. The Sudan III staining was more extensive in rabbits injected with saline than in those that received LDL. In addition, plasma cholesterol, plasma phospholipid, VLDL-cholesterol and LDL-cholesterol levels were lower in LDL-injected rabbits than in those injected with saline. But HDL-cholesterol levels were not significantly different between LDL-cholesterol and saline-injected rabbits. Although the exact cause of antiatherogenic effect of normal LDL is not clear, it seems reasonable to suggest that at least some normal LDL acts as a 'good' lipoprotein, namely antiatherogenic lipoprotein, in our experimental protocol.

Animals↗

Qualitative difference of subcellular localization of tumor-associated carbohydrate (Le(x)) antigens in renal cell carcinoma and normal kidney.

Renal cell carcinomas are immunohistochemically positive for oligosaccharides with the Le(x) determinant (Gal beta 1----4[Fuc alpha 1----3]GlcNAc) and its derivatives, as oncofetal antigens, and their expression is closely related to a better prognosis of the patients. This study was designed to clarify the difference in antigen localization at the ultrastructural level between renal cell carcinoma and normal tissues. In normal kidneys, Le(x) detected by monoclonal antibody (MAb) FH 2 and sialylated extended Le(x) (sialyl Le(x)-i) by MAb FH 6 were identified along the plasma membrane of microvilli of proximal tubule epithelial cells, with occasional immunoreactivity along the basolateral plasma membranes. Intracellular localization was very sparse. Renal cell carcinoma showed localization of Le(x) and sialyl Le(x)-i antigens along the cell membrane and in the cytosol as aggregates or filaments. Immunoreactive materials were also observed in the lumen formed among carcinoma cells. The cytosolic immunoreactivity, not observed in the normal kidney, was regarded as "abnormal cytosolic accumulation" of the antigens. This pattern was more pronounced in clear-cell carcinoma. Pretreatment of specimens with chloroform-methanol, which extracts glycolipids, decreased immunoreactivity in carcinoma tissues, particularly that in the cytosol. The extracts contained substances immunoreactive for MAb FH6. Our study has demonstrated that (a) remarkable changes occur in the ultrastructural localization patterns of sialyl Le(x)-i and Le(x) in renal cell carcinoma and (b) considerable amounts of glycolipids are contained in the substances with sialyl Le(x)-i deposited in the cytosol of clear-cell carcinoma.

Adenocarcinoma↗