Biomedical subjects
Publications and source records attributed to S Fujii.
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Cell cycle regulatory molecules were analysed in normal human endometrial tissue using antibodies against cyclins D1, E, A, and B1 and cyclin-dependent kinases (CDKs) cdk4, cdk2, and cdc2. The expression of these regulatory molecules in gland cells and stromal cells was compared with the expression of oestrogen receptors (ER), progesterone receptors (PR), and Ki67 (a growth-related molecule). In general, a substantially higher percentage of the gland cells stained positive for cyclins and CDKs during the proliferative phase of the menstrual cycle. Cyclin E, cdk2 and/or cdk4 were especially apparent in the cytoplasm of most of the gland cells as well as in the stromal cells. In contrast, most of the regulatory molecules were undetectable in the gland cells by the end of the secretory phase of the cycle, but they did not decline in the stromal cells. The data also revealed that ER, PR, and Ki67 in both gland cells and stromal cells follow the same basic pattern of expression as the cyclins and CDKs. These results suggest that cyclins and CDKs are functionally involved in the rhythmic proliferation of normal human endometrial tissue, and the action of these agents may be related to the endometrial levels of sex steroids and Ki67.
To clarify the biological characteristics of uterine leiomyomas, cells explanted and cultured from uterine leiomyomas and from normal myometrial tissue were observed by time-lapse cinemicrography and phase-contrast microscopy. The histological characteristics were evaluated by electron microscopy and immunofluorescence microscopy, and these observations revealed significant differences. By time-lapse cinemicrography, the cells cultured from leiomyomas and myometrium differed in their behaviour. Cells from the myometrium started to grow in parallel with the cell's major axis and formed topographically uniform hills and valleys by day 21 of culture. In contrast, the cells from leiomyomas started to grow irregularly, as if having no contact inhibition, and formed ball-like aggregates of cells by day 21 of culture. The aggregates resembled the nodules of leiomyoma in vivo. Ultrastructurally, cells from both leiomyomas and myometrium had typical features of smooth muscle. Immunofluorescently, a different distribution of alpha-smooth muscle actin-positive filaments and different staining of cellular fibronectin and N-cadherin between the cells from leiomyomas and myometrium were observed, which may contribute in part of the different behaviour of the cells. Given that the explant cell culture system resembles the features of uterine leiomyomas in vivo, this suggests that it can be used as an in-vitro model.
An assay method for GDP-L-Fuc:N-acetyl-beta-D-glucosaminide alpha 1-6fucosyltransferase (alpha 1-6FucT; EC 2.4.1.68) activity has been developed, involving a fluorescent pyridylaminated substrate. A glycopeptide derived from bovine gamma-globulin was coupled with 4-(2-pyridylamino)butylamine (PABA) through the peptide bond, and the following substrate was obtained. [equation: see text] The substrate and guanosine diphospho-fucopyranoside (GDP-Fuc) were incubated with a crude enzyme extract for 2 h, and then the enzymatic product was separated by reversed phase HPLC. Quantitation of the product involved measurement of the fluorescence intensity of the fucosylated pyridylaminated sugar. The structures of both synthesized GnGn-bi-Asn-PABA (substrate), and synthesized GnGnF-bi-Asn-PABA (product) were analyzed by 1H NMR. The enzymatic product was also analyzed by 1H NMR and was found to have alpha 1-6fucose at the reducing end GlcNAc. This method is highly specific for alpha 1-6FucT and is applicable for various experiments, including purification and cell culture ones.
Histological features of 21 cases of postmenopausal endometriosis as well as the corresponding eutopic endometrium were studied. Immunocytochemical analysis of estrogen receptor (ER), progesterone receptor (PR), and Ki-67 nuclear antigen was also performed on the endometriotic lesions as well as the corresponding endometrium. All the cases studied were complicated by various other gynecologic diseases, making the preoperative diagnosis of endometriosis unlikely. Histologically, the endometriotic foci contained both epithelial and stromal components mostly lacking atrophic features in contrast to the corresponding eutopic endometrium. The endometriotic lesions and the corresponding endometrium exhibited immunoreactivity with ER in both the glands and stroma. The stromal cells of endometriotic lesions revealed positive PR staining, whereas those of the eutopic endometrium were either negative or weakly positive for PR, but the epithelial components of both were positive for PR. The positivity of Ki-67 antigen tended to be higher in the stroma of the postmenopausal endometriosis than in the eutopic endometrial stroma. Endometriotic lesions appear to remain biologically active, with proliferative activity as well as retained hormonal responsiveness, even in the low-estrogen milieu of the postmenopausal age.
OBJECTIVE: We describe the technique of cosmetic reconstruction after mastoidectomy in the transpetrosal-presigmoid approach for petroclival lesions. TECHNIQUES: This technique involves raising a single temporo-occipital bone flap and cosmetic mastoidectomy, removing the mastoid bone fragments (by means of a rongeur instead of an air drill) for later reconstruction. Replacement of the bone fragments mixed with fibrin glue enables tamponade against the dura to be achieved without a fat graft. RESULTS: Computed tomographic scans and plain cranial films taken a few years later showed successful reconstruction of the mastoid bone with a good appearance of the retromastoid area in seven patients who underwent this procedure. The only complication was infection in one patient from cerebrospinal fluid leakage from the wound. CONCLUSION: We have developed a simple and easy technique of cosmetic reconstruction after the transpetrosal-presigmoid approach.
BACKGROUND: Pathogenetic effects of active immune cell products on the coagulation and fibrinolytic system proteins in liver and endothelial cells--primary sites of synthesis of these proteins--have not been elucidated. METHODS: We incubated highly differentiated human hepatoma cells (Hep G2) and human umbilical vein endothelial cells (HUVECs) for 24 h with recombinant human leukaemia inhibitory factor (LIF) and oncostatin M (OSM)--cytokines that are implicated in acute phase reactions during inflammation and which bind to the same cell surface receptor unit (glycoprotein gp 130). LIF was also given to mice in vivo. Contents of coagulation and fibrinolytic system proteins in cell culture supernatants and in mouse liver were determined. RESULTS: LIF increased the accumulation of urokinase plasminogen activator (u-PA) in the Hep G2 cell culture supernatants determined by enzyme-linked immunosorbent assay (ELISA) (0.21 +/- 0.03 (SE) ng/ml at baseline; 0.40 +/- 0.05 ng/ml at 100 U/ml, P < 0.05; 0.57 +/- 0.06 ng/ml at 500 U/ml, P < 0.01; n = 9) without altering total protein content. OSM elicited a similar effect (0.25 +/- 0.04 ng/ml at baseline, 0.62 +/- 0.19 ng/ml at 1 ng/ml; P < 0.05, n = 6). A monoclonal antibody against gp 130 abrogated the response to both agents (n = 9). Plasminogen activator inhibitor type-1 (PAI-1) (assayed by ELISA, n = 9), the PAI-1 binding protein, vitronectin (immunoprecipitation, n = 3) and tissue factor (ELISA, n = 3) were not affected by LIF, but fibrinogen production increased up to twofold with LIF (500 U/ml; Western blot, n= 3). In HUVECs, synthesis of tissue type plasminogen activator or PAI-1 was not altered by LIF or OSM (ELISA, n = 9). In vivo, intraperitoneal recombinant murine LIF (2 mu g) increased liver concentrations of u-PA by 30% and fibrinogen by 220% in mice. CONCLUSIONS: LIF and OSM produced by immune cells may modify fibrinolysis and coagulation by altering expression of u-PA and fibrinogen, thereby contributing to coagulopathy.
We analyzed deoxyribonucleic acids from blood samples of five Japanese von Hippel-Lindau (VHL) disease families (three familial cases, two new mutations) for the presence of VHL gene mutations by single-strand conformational polymorphism analysis and direct sequencing. Four of the five families showed germ line mutations in VHL gene, comprising 2 missense mutations, 1 deletion, and 1 splice-site mutation. Two families had VHL gene mutations at exon 1; 1 family at exon 3; and 1 family at the splice-site adjacent to exon 3. Presymptomatic patients were accurately diagnosed by these methods. However, one family did not show a VHL gene mutation in the germ line but showed a somatic mutation at exon 2 in the hemangioblastoma tissue. The consequence of the somatic mutation was a microdeletion leading to a frameshift mutation. Our study is the first report of VHL gene analyses of Japanese VHL disease families, and suggests that not only germ line mutation, but also somatic mutation can lead to development of a tumor associated with the VHL disease.
Primary lung adenocarcinomas in non-smoking females are increasing in the USA and Japan. Environmental factors such as passive smoking, asbestos, domestic radon, and hormonal effects have been implicated, but the etiology is still uncertain. We therefore analyzed point mutations of p16 gene, a newly characterized tumor suppressor gene, and compared the results with alterations of p53 gene in 28 primary lung adenocarcinomas in non-smoking Japanese females. There were no cases with somatic point mutation of p16 gene, except for one case with two germline mutations (silent mutations). In contrast, six out of 16 informative cases showed loss of heterozygosity of p53 gene using a TP53 microsatellite marker and 19 out of 28 cases showed expression of oncoprotein using DO-7 immunohistochemistry. These findings suggest that p16 gene alteration is a rare event in primary lung adenocarcinomas in Japanese non-smoking females, compared with alterations of the p53 gene.
OBJECTIVES: To verify the pattern of the spread of tumors in ovarian dysgerminoma, with special reference to intraperitoneal metastasis, and to assess clinicopathologic factors for predicting tumor extension. METHODS: Detailed data regarding ascitic cytology, macroscopical findings at surgery, and the histopathology of the surgical specimens were retrospectively reviewed in 12 patients with dysgerminoma who were treated at Kyoto University Hospital. The relationships between the tumor extension and the period of symptoms, the serum lactic dehydrogenase (LDH) level, and the operative findings also were analyzed. RESULTS: Ascitic cytology revealed a high incidence of positivity in 6 of the 10 (60%) cases examined. Extraovarian metastases were present in 4 of the 6 (67%) cases with positive cytology, and in 1 of the 4 (25%) cases with negative cytology. Intraperitoneal metastatic nodules were detected in 5 of the 12 (42%) patients either by inspection during surgery or by postsurgical histological examination. In addition, these metastatic lesions were 5 or fewer in number and 7 mm or less in diameter, except in 1 patient with widespread disease. The presence or absence of extraovarian spread of the tumor was not significantly correlated with the period of symptom, the serum LDH level, the size of the primary tumor, or the volume of the ascitic fluid. CONCLUSION: The incidence of intraperitoneal spillage and/or metastases of dysgerminoma cells might be higher than previously reported. These findings indicate the importance of ascitic cytology and careful inspection at the time of operation, as well as the rationale of postsurgical chemotherapy for dysgerminoma of an apparently early stage.
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Several saccharides were found to be significantly effective in providing protection against hydrostatic pressure and high temperature damage in the yeast Saccharomyces cerevisiae. The extent of barotolerance and thermotolerance with seven different sugars showed a linear relationship to their mean number of equatorial OH groups. The same linear relationship is seen when sugars protect protein molecules against elevated temperatures in vitro. Some sugars were more effective in providing protection against hydrostatic pressure nearly a hundred times than high temperature. Pre-heat shock treatment on yeast cells induce various stress tolerances. In this report, pre-heat shocked cells showed potent protection against elevated temperature, but these cells showed faint protection against elevated pressure. These results suggest that sugars may protect cells against hydrostatic pressure and high temperature in a similar manner, probably by stabilizing the macromolecule(s), and such type of protection may be suited for pressure stress.
Restenosis after balloon dilitation of atherosclerotic arteries reflects migration and proliferation of vascular smooth muscle cells and infiltration of monocyte/macrophages. Hypercholesterolemia may contribute to this phenomenon. Accordingly, we used the lipid-lowering agent gemfibrozil to determine whether potentially detrimental effects of hypercholesterolemia on vascular remodeling after mechanical injury could be attenuated. New Zealand white rabbits fed either a chow diet (control), a 0.25% cholesterol-enriched diet, or a 0.25% cholesterol-enriched diet supplemented with gemfibrozil (0.05%, 0.1%, or 0.02%) for one week were subjected to balloon-induced carotid injury and maintained on the same diet for an additional 4 weeks. Histology of the vascular wall was then characterized. Plasma triglycerides before and 4 weeks after injury did not change in any of the treatment groups (p = 0.24). Plasma cholesterol increased in all animals receiving the high cholesterol diet, and the increases remained unaffected by supplementation with gemfibrozil. In control rabbits, intimal thickening area [intima (mm2)/(intima + media (mm2))] 4 weeks after injury was 27.0 +/- 7.7% (n = 16). Values were the same in hypercholesterolemic rabbits (29.7 +/- 11.8%, n = 12; p = ns). However, in 16% the lumen was completely occluded by thrombus and intimal thickening could not be quantified. In hypercholesterolemic rabbits given gemfibrozil, intimal thickening was increased by 33% compared with controls (35.9 +/- 11.6%, n = 39, pound 0.05) and by 21% compared with hypercholesterolemic animals not given gemfibrozil (p = ns). None had thrombotic luminal occlusion. Macrophages detected immunohistochemically were only modest in number in vessels from control animals. In vessels from hypercholesterolemic animals and from animals whose diets were supplemented with gemfibrozil, macrophages were increased in number in both intima and media. Thus, gemfibrozil did not appear to attenuate processes implicated in restenosis. Its attenuation of thrombotic occlusion may be related to effects we have noted it exerts on fibrinolytic systems independent of lipid metabolism.
A study of fertility and fetal development of prulifloxacin, a new antibacterial agent, was conducted in Sprague-Dawley rats. Male rats were given the drug orally from 63 days before mating to the end of the mating period. Female rats were given from 14 days before mating to day 7 of pregnancy. The dose levels for both males and females were 0 (control), 30, 170 and 1000 mg/kg. The females were sacrificed on day 20 of pregnancy for examination of their fetuses. In the 170 mg/kg group, water consumption increased in the female rats, and food consumption decreased and the cecum increased in weight and was enlarged in the males and the females. In the 1000 mg/kg group, body weight gain was suppressed in the males, and food consumption decreased, water consumption increased and cecum increased in weight and enlarged in the males and the females. Moreover, this dose caused white spots and rough surface of the kidney in the males. Prulifloxacin had no adverse effects on reproductive function of the parent animals, and also on development of the fetuses. These results show that the NOAEL of prulifloxacin are 30 mg/kg for general toxicity in parent animals, 1000 mg/kg for reproductive function of parent animals and for development of fetuses.
A study of the effect of prulifloxacin, a new antibacterial agent, during the period of organogenesis was conducted in Sprague-Dawley rats. Female rats were given prulifloxacin orally at dose levels of 0 (control), 30, 300, and 3000 mg/kg from day 7 to day 17 of pregnancy. Twenty-four or twenty-five female rats per dose level were sacrificed on day 20 of pregnancy for examination of their fetuses, and the pregnant rats (12-15 per dose levels) were allowed to deliver naturally for postnatal examination of their offspring. In the 300 or 3000 mg/kg groups, the food consumption decreased and water consumption increased in the dams. In the 3000 mg/kg group, the body weight gain was suppressed in the dams. In the 300 and 3000 mg/kg groups, the weights of cecum increased and the enlargement of cecum was observed. In the 3000 mg/kg group, white spots and rough surface of the kidney and renal tubular nephrosis with crystalline substance were observed and the weight of the kidney increased in the dams, and the body weight decreased and retarded ossifications in the fetuses. Prulifloxacin had no effect on the number of corpora lutea and implantations, or on fetal mortality, sex ratio, or external and visceral development of the fetuses. Prulifloxacin also did not affect delivery and the number of live newborns and birth index, or have any adverse effects on the postnatal development of the first generation offspring (F1) such as differentiation, functional development, emotionality, motor ability, learning ability or reproductive performance. Prulifloxacin also had no adverse effects of the second generation offspring (F2). These results show that the NOAEL of prulifloxacin are 30 mg/kg for general toxicity in mother animals, 3000 mg/kg for pregnancy and delivery of mother animals and 300 mg/kg for development of their offspring.
A study of the effect of prulifloxacin, a new antibacterial agent, during the period of organogenesis was conducted in New Zealand White rabbits. Female rabbits were given prulifloxacin orally at dose levels of 0 (control), 10, 30 and 100 mg/kg from day 6 to 18 of pregnancy. Female rabbits were sacrificed on day 29 of pregnancy for examination of their fetuses. In the 30 mg/kg group, food and water consumption decreased. In the 100 mg/kg group, soft stool was observed and body weight gain, food consumption and water consumption decreased. Premature delivery (2/16) occurred and enlargement of cecum and increased weight of cecum were observed. The number of fetal death increased in the 100 mg/kg group. However, prulifloxacin had no effects on the number of corpora lutea, implantations and live fetuses, and on body weight, placental weight, sex ratio, and external, visceral and skeletal development of live fetuses. These results show that the NOAEL of prulifloxacin are 10 mg/kg for general toxicity in mother animals, 30 mg/kg for pregnancy of mother animals and for development of fetuses.
A peri- and postnatal study of prulifloxacin, a new antibacterial agent, was conducted in Sprague-Dawley rats. Female rats were given prulifloxacin orally at dose levels of 0 (control), 30, 100 and 300 mg/kg from day 17 of pregnancy to day 21 after delivery. All pregnant rats were allowed to deliver naturally for postnatal examination of their offspring. Food consumption in the period of pregnancy decreased in the dams given 100 and 300 mg/kg, and food and water consumption during lactational period increased in the dams given 300 mg/kg. The weight of the cecum of the dams increased in 100 and 300 mg/kg groups, and these doses caused enlargement of the cecum. Prulifloxacin had no effects on general sign, delivery, nursing, body weight gain, and kidney. Prulifloxacin also did not affect the number of live newborns and birth index, and have any adverse effects on the postnatal development of the first generation offspring (F1) such as differentiation, functional development, emotionality, motor ability, learning ability and reproductive performance. Prulifloxacin also had no adverse effects on the second generation offspring (F2). These results show that the NOAEL of prulifloxacin are 30 mg/kg for general toxicity in mother animals, 300 mg/kg for pregnancy and delivery of mother animals and development of their offspring.
The life cycle of Arabidopsis plants was examined by growing them on a horizontal clinostat. Seeds on agar media were allowed to germinate and seedlings were grown under a simulated microgravity on a horizontal clinostat. Clinorotation (3 rpm) did not appear to interfere with germination of plant seeds and development of cotyledons and leaves. Stress relaxation parameters of the cell wall, the minimum relaxation time and the relaxation rate did not appear to be affected by clinostat rotation. On the other hand, the length of inflorescences was reduced to 61-62% by clinostat rotation. Rotation was found to inhibit the polar transport of auxin, although inflorescence growth and auxin transport were not completely inhibited. From these facts, it is possible that the life cycle in Arabidopsis plants could be accomplished in space, although growth phenomena involving auxin transport and its action may be disturbed. Plants may have a capacity to grow in space and we may be able to cultivate crops in space.