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Biomedical subjects

M Sakai

Publications and source records attributed to M Sakai.

At least 199 records · Page 11Linked to original sources
PubMed1998

Flow cytometric analysis of DNA content and Ki-67-positive fractions in the diagnosis of salivary gland tumors.

To explore the utility of flow cytometry (FCM) for the diagnosis of histopathology of salivary gland tumors, fresh materials taken at surgery from 23 Warthin's tumors, 57 pleomorphic adenomas, and 14 malignant tumors were analyzed for DNA ploidy and proliferative cell activities, including S-phase fraction (SPF), G2- plus M-phase fraction (G2M), and Ki-67-positive fraction. To facilitate this study, glands were taken from all major salivary sites and minor glands in the head and neck. DNA aneuploidy was not detected in the benign tumors. Nine of 14 malignant tumors showed DNA aneuploidy. The percentage of SPF or G2M of the malignant tumors was significantly higher than those of the benign tumors. The percentage of Ki-67-positive fraction of pleomorphic adenomas was comparable to that of malignant tumors and was significantly higher than that of Warthin's tumors. Ki-67 of 20% as a cut-off had a sensitivity of 88%, specificity of 100%, and accuracy of 91% for differentiating pleomorphic adenomas from Warthin's tumors. In analyzing DNA content and proliferative activities by FCM, we could distinguish among the three major histopathologies of salivary gland tumors. Warthin's tumors showed low SPF + G2M with low Ki-67, pleomorphic adenomas had low SPF + G2M with high Ki-67, and malignant tumor showed high SPF + G2M with high Ki-67. The high percentage of the Ki-67-positive fraction seen in pleomorphic adenomas may reflect their potential biological aggressiveness manifested as tumor recurrence or malignant transformation.

Adenolymphoma↗
PubMed1998

Effects of absorption enhancers on cytoskeletal actin filaments in Caco-2 cell monolayers.

Histomorphological changes of actin filaments, intracellular levels of calcium ion, and amount of released lactate dehydrogenase (LDH) were examined, in order to elucidate the mechanism of action of three absorption enhancers, i.e., sodium caprate (Cap-Na), sodium deoxycholate (Deo-Na), and dipotassium glycyrrhizinate (Grz-K), using Caco-2 cell monolayers. The structure of actin filaments in microvilli was slightly modified by 0.5 %(w/v) Grz-K and was significantly changed by 0.2 %(w/v) Cap-Na, 0.05 %(w/v) Deo-Na, and 0.0015 %(w/v) ionomycin. All of the enhancers, except Grz-K, induced significantly histomorphological changes in the actin filaments on the middle depth and basal side of the cells. Furthermore, the altered structure of the actin filaments in the monolayers was restored after removal of the Cap-Na, Grz-K and ionomycin, but not Deo-Na. Intracellular levels of calcium ion increased in the following order: ionomycin = Cap-Na > Deo-Na. However, the intracellular calcium ion levels decreased by treatment with Grz-K. The changes in transepithelial electric resistance (TEER) at the initial stage of treatment with all enhancers correlated with intracellular calcium ion levels. These results suggest that one of the mechanisms by which these agents exert absorption-enhancing activity involves structural alterations in the cytoskeletal actin filaments which are provoked by changes in intracellular calcium ion levels. Only the monolayers which were treated with 0.05 %(w/v) Deo-Na released a significant amount of LDH and irreversibly altered the structure of actin filaments, thus indicating that Deo-Na might affect the actin filaments not only by increasing intracellular calcium ion level but also by other, presently unknown factors.

Actins↗
PubMed1998

Cochlear otosclerosis 30 years after stapedectomy confirmed by CT, MRI.

A 64-year-old Japanese male patient, status post-stapedectomy, presented with progressive, bilateral SNHL of 30 years' duration. CT showed extensive demineralization of the otic capsule, suggestive of otospongiotic change in both ears. There were no hyperostotic lesions found, which would have been suggestive of otosclerotic change. MRI showed a morphologically intact membranous labyrinth in both ears, with no obstruction of the cochlear lumen. It was concluded that the patient has progressive otospongiotic change of the otic capsule, but no otosclerotic change.

Calcification, Physiologic↗
PubMed1998

Co-expression of hepatocyte growth factor and its receptor in human prostate cancer.

Hepatocyte growth factor acts differently depending on the organs or tumours involved. It may be produced simultaneously with its receptor, c-Met, in several types of malignant tumour cells and may exercise an autocrine regulation. To analyse the effect of hepatocyte growth factor in human prostate cancer, we conducted immunohistochemistry, in situ hybridization and the reverse transcriptase polymerase chain reaction. The first two techniques revealed the growth factor in prostate cancer cells, and the polymerase chain reaction confirmed this expression. c-Met is expressed in prostate cancer cells, but not in interstitial cells. Hepatocyte growth factor is expressed in interstitial cells, especially in hormone-treated cancer tissue, indicating that the growth factor pathway changes with the hormonal status. Low-grade tumours expressed c-Met at the plasma membrane. Higher grade tumours tended to express it in the cytoplasm, suggesting that the role of c-Met as the hepatocyte growth factor receptor was blocked in higher grade tumours. The relationship between the growth factor and its receptor is thus influenced by hormonal status and differentiation in prostate cancer and is not explained simply in terms of autocrine or paracrine action.

Aged↗
PubMed1998

Stimulation of nitric oxide release from rat spinal cord by prostaglandin E2.

1. We recently demonstrated that intrathecal administration of prostaglandin E2 (PGE2) and PGF2alpha induced allodynia through a pathway that includes the glutamate receptor and nitric oxide (NO)-generating systems from pharmacological studies. In order to clarify the involvement of NO in prostaglandin-induced allodynia, we measured NO released from rat spinal cord slices by a chemiluminescence method. 2. PGE2 stimulated NO release from both dorsal and ventral regions all along the spinal cord. PGE2 stimulated the release within 10 min and increased it in a time-dependent manner. 3. The PGE2-induced NO release was observed at 100 nM-10 microM. PGF2alpha stimulated the release at concentrations higher than 1 microM, but PGD2 (up to 10 microM) did not enhance it. 4. 17-Phenyl-omega-trinor PGE2 (EP1 > EP3) and sulprostone (EP1 < EP3) were as potent as PGE2, but PGE1 was less potent, in stimulating NO release. While M&B 28767 (EP3) did not enhance the release, butaprost (EP2) stimulated it at 1 microM. The PGE2-evoked release was blocked by ONO-NT-012, a bifunctional EP1 antagonist/EP3 agonist. 5. The PGE2-evoked release was Ca2+-dependent and blocked by MK-801 (NMDA receptor antagonist) and L-NAME (NO synthase inhibitor). The release was also inhibited by PGD2 and dibutyryl-cyclic AMP. 6. The present study demonstrated that PGE2 stimulates NO release in the rat spinal cord by activation of NMDA receptors through the EP1 receptor, and supports our previous findings that the NO-generating system is involved in the PGE2-induced allodynia.

Animals↗
PubMed1998

Increased nitric oxide in nasal lavage fluid and nitrotyrosine formation in nasal mucosa--indices for severe perennial nasal allergy.

BACKGROUND: The nose contributes the large amount of nitric oxide (NO) to exhaled air. NO is a mediator of vasodilation and yields peroxynitrite (ONOO-) by reacting with superoxide (O2-). ONOO attacks tyrosine residues to form nitrotyrosine. OBJECTIVE: The aim of this study was to examine the pathophysiological role of NO in nasal mucosa in patients with perennial nasal allergy. METHODS: We measured nitrite and nitrate (NO2-/NO3-) and 3',5'-guanosine monophosphate (cyclic GMP) in nasal lavage fluid, and also measured haemoglobin concentration in nasal mucosa as an indicator of blood volume in the patients and healthy volunteers. The deleterious role of NO was also investigated by measuring nitrotyrosine in nasal mucosa. RESULTS: The NO2-/NO3- concentration in the nasal lavage fluid was 39.5+/-2.8 microM in healthy volunteers (n=40), 42.4+/-3.0 microM in patients with mild allergy (mild group, n=32), and 88.7+/-6.6 microM in patients with severe allergy (severe group, n=61). In the patients whose symptoms were improved with treatment, NO2-/NO3- levels decreased to 45.7+/-10.4 microM. The concentration of cyclic GMP in nasal lavage fluid was higher in the severe group than in the healthy volunteers. The mucosal haemoglobin index was 88+/-4 in the healthy volunteers, 67+/-4 in the mild group, and 53+/-2 in the severe group. The formation of nitrotyrosine was expressed 0.58+/-10% to total tyrosine in the severe group (n=11), but was not found in non-allergy patients (n=9). CONCLUSIONS: The production of NO was increased in patients with perennial nasal allergy, but the blood flow in the nasal mucosa of patients was reduced. Nitrotyrosine formation suggests that there is a process of ONOO(-)-induced damage in mucosa of patients with the perennial nasal allergy and this damage may limit the dilatation of blood vessels, despite the presence of excessive NO.

Adult↗
PubMed1998

The treatment of neuroleptic malignant syndrome using dantrolene sodium.

The use of dantrolene sodium (DS) in the treatment of neuroleptic malignant syndrome (NMS) was studied from the data of 21 cases of NMS, which satisfied the diagnostic criteria of Pope et al. The mean dosage of DS was 0.97+/-0.52 mg/kg per day and the mean duration of treatment was 8.3+/-5.3 days. It was confirmed that the mean duration of treatment with DS tended to be longer in the cases involving disturbance of consciousness. However, there were no significant differences in the duration of treatment or the mean daily dosage, in the cases without disturbance of consciousness regardless of any other complications and symptoms. Also, there were no significant differences in the mean duration of treatment using DS and the mean dosage of DS between five cases that were considered to be 'typical' and 16 cases that were considered to be 'incomplete'. In addition, the prognoses of these cases were good such that not even one death occurred. In 13 cases, the treatment of psychiatric symptoms with thioridazine was started without recurrence of NMS.

Adolescent↗
PubMed1998

Serotonin enhances the production of type IV collagen by human mesangial cells.

BACKGROUND: The plasma concentration of 5-hydroxytryptamine (5-HT) in diabetic patients is higher than that in normal subjects. Since recent reports have demonstrated the presence of 5-HT2A receptor in glomerular mesangial cells, it is possible that 5-HT may be involved in the development of diabetic nephropathy through the 5-HT2A receptor in mesangial cells. Because expansion of the glomerular mesangial lesion is a characteristic feature of diabetic nephropathy, we examined the effect of 5-HT on the production of type IV collagen by human mesangial cells. METHODS: Human mesangial cells were incubated with 5-HT with or without 5-HT receptor antagonists, protein kinase C (PKC) inhibitor or transforming growth factor-beta (TGF-beta) antibody. Type IV collagen mRNA and protein concentration in medium were measured by Northern blot analysis and enzyme-linked immunosorbent assay (ELISA), respectively. TGF-beta mRNA and bioactivity in the medium were measured by Northern blot analysis and bioassay using mink lung epithelial cells, respectively. RESULTS: 5-HT stimulated the production of type IV collagen by human mesangial cells, which was inhibited by ketanserin and sarpogrelate hydrochloride, 5-HT2A receptor antagonists, but not by ondansetron, a 5-HT3 receptor antagonist. 5-HT increased the bioactivities of both active and total TGF-beta. However, the 5-HT-enhanced production of type IV collagen was completely inhibited by an anti-TGF-beta antibody. Furthermore, a PKC inhibitor, calphostin C, inhibited the 5-HT-induced increase in type IV collagen secretion, and the activity of membrane PKC was increased by 5-HT. Phorbol ester activated type IV collagen production as well as active and total TGF-beta. Calphostin C completely inhibited the 5-HT-enhanced activity of active TGF-beta, but did not inhibit exogenous TGF-beta-induced increase in type IV collagen secretion. CONCLUSIONS: Our results suggest that 5-HT-enhanced production of type IV collagen by human mesangial cells is mediated by activation of PKC and subsequent increase in active TGF-beta activity.

Cells, Cultured↗
PubMed1998

Differential developmental process of respective behavioral states in human fetuses.

The presence of behavioral states has been recognized in human fetuses. However, developmental process of fetal behavioral states remains to be clarified. In thirty-one normal pregnant women between 20 and 41 weeks' gestation, fetal movements were examined by using multiple ultrasonic machines. The entire period was divided into eye movement (EM), no eye movement (NEM), and unclassified periods according to the mode of appearance of rapid eye movements. We investigated changes in the ratio of EM and NEM periods and the incidence of various fetal movements in these periods. The ratio of EM periods increased stepwise between 20 and 32 weeks. The difference in the incidence of "startle" and jaw movements between EM and NEM periods was found at 24-27 and 32-35 weeks, respectively. We found a significant decrease in the incidence of total body movements exclusively in NEM periods at 36-39 weeks and a significant increase in the incidence of "rolling" in EM periods at 40-41 weeks. Eye opening was ultrasonographically recognized, providing evidence for awake state in the human fetus. This study revealed the characteristic appearance of fetal movements as related to the presence or absence of EM during the second half of pregnancy, and thereby pointed to differential development process of behavioral states of human fetuses.

Adult↗
PubMed1998

Tumor necrosis factor-alpha regulates the gene expression of macrophage migration inhibitory factor through tyrosine kinase-dependent pathway in 3T3-L1 adipocytes.

Macrophage migration inhibitory factor (MIF) has been rediscovered as a proinflammatory cytokine, pituitary hormone, and glucocorticoid-induced immunoregulator. We have recently identified the expression of MIF in adipocytes and found that tumor necrosis factor (TNF)-alpha stimulates its secretion from 3T3-L1 adipocytes. Since adipocytes are regarded as a potential source of various biologically active substances, we examined in more detail the effect of TNF-alpha on MIF expression in 3T3-L1 adipocytes in the present study. We found that TNF-alpha induced MIF mRNA in dose- and time-dependent manners. After stimulation with TNF-alpha, the amount of intracellular MIF protein was unchanged or slightly decreased, concomitant with increased release of this protein into the extracellular space. This observation indicates that TNF-alpha stimulates MIF secretion from the constitutively expressed intracellular pool of 3T3-L1 adipocytes and promotes de novo synthesis of MIF. From evaluation of the mechanism of MIF gene expression, we found that tyrosine kinase inhibitors, either genistein or herbimycin A, suppressed the MIF mRNA induction by TNF-alpha. The results suggest the possibility that upregulation of MIF mRNA expression by TNF-alpha is mediated by a tyrosine kinase-dependent pathway. Taken together, the present observations shed light on the role of MIF in the metabolism of obesity and diabetes.

3T3 Cells↗
PubMed1998

Bile acids influence hepatic chemiluminescence in normal and oxidative-stressed rats.

The aim of the present study was to clarify whether bile acids influence chemiluminescence (CL) in the liver in vivo. Hepatic CL was determined on the surface of the liver of anaesthetized rats by using a photon counter. In normal rats, hepatic CL was significantly decreased 30 min after enteral administration of chenodeoxycholic acid (CDCA) or deoxycholic acid (DCA), but returned to its initial level 3 h later, after part of the CDCA administered was metabolized. Ursodeoxycholic acid (UDCA) and cholic acid had no effect on CL. In contrast, hepatic CL was markedly increased 30 min after CDCA or DCA administration in rats given either buthionine sulphoximine (BSO), an inhibitor of gamma-glutamylcysteine synthetase, or diethyldithiocarbamate (DDC), an inhibitor of both superoxide dismutase and glutathione peroxidase. Chenodeoxycholic acid further increased the CL of BSO- or DDC-treated rats during inhalation of oxygen via a tracheal cannula. Coadministration of UDCA eliminated the effects of CDCA on the hepatic CL of normal and BSO- or DDC-treated rats with or without oxygen inhalation. We conclude that cytotoxic bile acids, such as CDCA, increase CL in the antioxidants-depleted or oxidative-stressed liver in vivo, but that UDCA prevents CDCA from developing CL.

Animals↗
PubMed1998

Cytotoxicity of absorption enhancers in Caco-2 cell monolayers.

This study was performed to evaluate the utility of absorption enhancers with reference to mucosal cell cytotoxicity. Overall assessment of the damage to plasma, lysosomal and nuclear membranes by three absorption enhancers, sodium deoxycholate, sodium caprate and dipotassium glycyrrhizinate, was performed on Caco-2 cell monolayers. The cytotoxicities of sodium deoxycholate (0.02-0.1% w/v), sodium caprate (0.1-0.5% w/v) and dipotassium glycyrrhizinate (0.5-2% w/v) were evaluated by the trypan blue-exclusion test, the protein-release test, the neutral-red assay, the DNA--propidium iodide staining test and the test for recovery of transepithelial electrical resistance (TEER) up to 24 h after treatment with each enhancer. Sodium dodecyl sulphate (SDS; 0.1% w/v), a potent surfactant, was used as positive control. SDS at this level was significantly cytotoxic whereas dipotassium glycyrrhizinate was not cytotoxic in any tests. Results from the trypan blue-exclusion and protein-release tests showed that high concentrations of sodium caprate (0.5% w/v) and sodium deoxycholate (0.1% w/v) were significantly cytotoxic to the plasma membrane. The neutral-red assay, an indicator of damage to lysosomal membranes, revealed that 0.5% (w/v) sodium caprate had no effect whereas the uptake of neutral red was slightly increased by treatment with 0.1% (w/v) sodium deoxycholate, implying that the compound had cell-growth-enhancing activity. Nuclear-membrane damage, as evaluated by the DNA--propidium iodide staining test, was severe in cell monolayers treated with 0.5% (w/v) sodium caprate compared with that induced by 0.1% (w/v) sodium deoxycholate. In the TEER recovery test, TEER failed to recover 24 h after treatment with 0.5% (w/v) sodium caprate and 0.1% (w/v) SDS, but recovered after treatment with 0.1% (w/v) sodium deoxycholate. The recovery of TEER might be related to nuclear membrane damage and cell-growth-enhancing activity. These results indicate that of the three classes of enhancer, dipotassium glycyrrhizinate was not cytotoxic and that high concentrations of sodium caprate and sodium deoxycholate could damage plasma and nuclear membranes.

Caco-2 Cells↗
PubMed1998

Metabolic and endocrine responses to cold exposure in chronically incubated extrauterine goat fetuses.

To investigate developmental aspects of metabolic and endocrine responses to cold exposure in fetuses, we conducted experiments on six goat fetuses, three aged 95-116 d of gestation (dGA; group I), and three aged 122-134 dGA (group II), using an extrauterine fetal incubation system that provided arterio-venous extracorporeal membrane oxygenation (A-V ECMO). The fetuses were cannulated via the umbilical vessels, and their blood gas exchange was totally supported by A-V ECMO, while they were maintained in an isothermal incubator containing artificial amniotic fluid. After confirming that fetuses were in metabolically stable condition in the extrauterine incubation system, fetal core temperature was lowered by 2 degrees C over 2 h by decreasing the temperature of incubating fluid from 39.5 degrees C. During and after cold exposure, fetal heart rate and arterial blood pressure remained unchanged. We observed significant increases in oxygen consumption and plasma concentrations of norepinephrine, epinephrine, adrenocorticotropic hormone, and cortisol in group II but not in group I fetuses. In addition, based on regression analysis, maximal changes of these parameters during cold exposure were linearly correlated with gestational age significantly, and the regression lines were found to intersect the x (gestational age) axis at around 98-106 dGA. These results suggest that metabolic and endocrine responses to cold exposure develop with gestational age in the goat fetus, the responses being manifested around 100 dGA.

Animals↗
PubMed1998

[Reproductive and developmental toxicity study of sodium N-[2-[4-(2,2-dimethylpropionyloxy) phenylsulfonylamino] benzoyl] aminoacetate tetrahydrate (ONO-5046.Na) (3). Teratogenicity study in rabbits].

Teratogenicity of sodium N[2-[4-(2,2-dimethylpropionyloxy) phenylsulfonylamino]benzoyl] aminoacetate tetrahydrate (ONO-5046.Na), a novel inhibitor of human neutrophil elastase, was studied. ONO-5046.Na was injected intravenously at doses of 0, 7.5, 15 and 30 mg/kg/day to pregnant Kbl: NZW rabbits from day 6 to day 18 of pregnancy. All female rabbits were sacrificed on day 29 of pregnancy and their fetuses were examined. There were no clinical signs or death attributable to ONO-5046.Na. One dam in the control group and 3 dams in the 30 mg/kg/day group aborted. Body weight gain in the 15 and 20 mg/kg/day groups and food intake in the 30 mg/kg/day group were decreased during the administration period. These changes had recovered by the end of the study. Kidney weight was increased in the 30 mg/kg/day group. There were no effects of ONO-5046.Na in necropsy findings at cesarean section in dams at any dose levels. Developmental toxicity of ONO-5046.Na was not found at any dose levels. From these results, it is considered that the NOAEL of ONO-5046.Na is 7.5 mg/kg/day for pregnant animals and 30 mg/kg/day for fetuses.

Animals↗
PubMed1998

[Study of optokinetic nystagmus and spino-motor functions with cyclo-rotatory optokinetic stimulus].

It has been shown that the neural channels from the otolith organs and the vertical semicircular canals connect to the cyclo-rotatory oculomotor system. However, only a few studies have been conducted to examine the relationship between the cyclo-rotatory optokinetic (COK) and the vestibular inputs. Accordingly, we have analyzed the cyclo-rotatory oculomotor output evoked by a COK stimulus, in 30 subjects; 21 normal adults, 4 elderly persons, and 5 persons with unilateral vestibular dysfunction. In addition, the spinal motor output was simultaneously recorded and comparatively analyzed. The increase in COK nystagmus (COKN) was less than that in horizontal and vertical OKN. There was a significant decline in the maximum slow phase eye velocity in the elderly subjects as well as in the subjects with unilateral vestibular dysfunction, compared to that in the normal adult subjects. Regarding the spinal motor output, in the subjects with unilateral vestibular dysfunction, a pronouned increase in the sway distance from the center of gravity was found with COK stimulus forward the lesioned side. However, with the COK stimulus, there was no significant correlation between the output functions of the cyclorotatory oculomotor system and those of the spinal motor system.

Adult↗
PubMed1998

[Evaluation of optokinetic afternystagmus (OKAN) induced by cyclo-rotatory optokinetic stimulation (C-OKst)].

It is not known whether the optokinetic afternystagmus (OKAN) can be evoked by cyclo-rotatory optokinetic stimulation (C-OKst) or not. Accordingly, this study was designed to investigate the problem by using a rotating white dome with random dots. Optokinetic nystagmus (OKN) and OKAN in standing upright, supine, and prone positions were recorded and analyzed with an infrared-ray CCD camera and computed picture analyzer. During stimulation, rotatory OKN showed clearly, but there was no slow-rise component. The OKAN was continuously recorded and analyzed. The component of this OKAN was found only in the head-horizontal direction (yaw axis) in all different head positions. It appeared fairly constantly in the prone posture. The velocity storage mechanism (VSM) had to be charged by the C-OKst, but the OKAN (1st phase) appeared only in the yaw axis, not in the roll axis.

Adult↗