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Biomedical subjects

A Fukuda

Publications and source records attributed to A Fukuda.

At least 91 records · Page 5Linked to original sources
PubMed1997

Dopamine-denervation enhances the trophic activity in striatum: evaluation by morphological and electrophysiological development in PC12D cells.

To evaluate the possibility that dopamine (DA) denervation enhances the trophic activity in striatum, normal or DA-depleted striatal tissue extract (N- or L-extract, respectively) was obtained, and their trophic effects on PC12D cells were investigated from the viewpoints of differentiation using morphological and electrophysiological analyses. Treatment with N- or L-extract induced neurite outgrowth in a concentration-dependent manner, and induced the enlargement of cell size. These effects were stronger in L-extract than in N-extract. Cation currents were investigated in whole cell patch-clamp mode. Development of cation current started with delayed-rectifier type K+ current (IK) and transient type K+ current (IA), followed by Ca2+ current (ICa) and tetrodotoxin-sensitive Na+ current (INa). INa was expressed more frequently in L-extract treated cells than N-extract treated cells at D7-9. The larger IK amplitude in L-extract treatment at D7-9 seemed to be related to the expression of INa. Development of IA was similar at any stage for both treatments. ICa development started at D3-5 after treatments, and the amplitude and current density were similar in both treatments. ICa was strongly blocked by omega-conotoxin GVIA (3 microM), indicating that N-type channels were mainly expressed after treatments. The data suggests that L-extract has stronger effects to hasten the differentiation of PC12D cells than N-extract by promoting the neurite outgrowth, cell enlargement and expression of voltage-dependent cation channels, especially INa and IK.

Animals↗
PubMed1997

Acute 3-nitropropionic acid intoxication induces striatal astrocytic cell death and dysfunction of the blood-brain barrier: involvement of dopamine toxicity.

Mechanisms underlying the selective vulnerability of the lateral striatal area to the toxic effects of 3-nitropropionic acid (3-NPA) were investigated in rats. A single exposure to 3-NPA (20 mg/kg, s.c.) induced no deficits in behavior and histology, but subsequent injection produced motor symptoms, catalepsy, lip smacking, abnormal gait, paddling, rolling, opisthotonos, tremor, recombence, somnolence and so on, in 30% of the animals within a few hours. Diffusion-weighted magnetic resonance imaging of the brains revealed an area of high signal intensity in the bilateral striata. By this stage (within a few hours), striatal astrocytes had become swollen and disintegrated. Extravasation of immunoglobulin G was detected, indicating blood-brain barrier (BBB) dysfunction. Electron microscopy revealed edema and disorganization of structures inside the astrocytic end-feet around the branches of the lateral striatal artery. Neurons were less vulnerable than astrocytes to the 3-NPA injury. Treatment of the rats with D2 receptor agonist prior to exposure to 3-NPA attenuated the behavioral abnormalities and histological damage whereas pretreatment with D2 antagonist exacerbated these changes. The concentrations of extracellular dopamine (DA) and dihydroxyphenyl acetic acid (DOPAC) were both increased in rats exposed to 3-NPA. In vitro imaging of astrocytes revealed a progressive increase in [Ca2+]i after superfusion with 3-NPA, and the 'ceiling' level was maintained even after extensive washing. DA superfusion also increased the astrocytic [Ca2+]i and this increase was reversible. Data indicate that 3-NPA-induced striatal damage was associated with astrocytic cell death and dysfunction of the BBB. Intracellular edema and extreme Ca2+ overload induced by the toxin were further aggravated by an increase in the level of DA activity. These factors acting either singly or in combination may trigger astrocyte destruction.

Animals↗
PubMed1997

Topography of hypoxic injury proved by argyrophilia in postnatal rat brain.

The argyrophil III method, a new esterification-silver staining approach, was used to elucidate regional differences in the susceptibility of developing brain to hypoxic-ischemic (H-I) injury. We created a unilateral common carotid artery-ligation model with hypoxia (8% oxygen) in postnatal day (P) 7, P14 and P21 rats. The argyrophil (i.e., deteriorated) neurons were apparent in the ipsilateral hippocampus, cortex, and striatum in each age group. Argyrophil neurons exhibited some morphological signs of the "early phase" of injury preceding the loss of structure and/or cell death in the "late phase," as indicated by hematoxylin-eosin (H-E) staining. The argyrophil neurons were apparent as early as 12 hours after the insult, whereas the histological changes revealed by H-E staining were subtle. The early phase and late phase histological changes had a stereotyped pattern of appearance in all ages studied. However, the duration of H-I situation required to produce argyrophil cells differed according to age. The most resistive age was P14 (P14 > P7 > P21) in this observation. Therefore, argyrophil III staining is feasible for H-I brain damage model in neonates. The results suggest that both the early phase and the late phase pathological processes after H-I injury have a characteristic topographical vulnerability that does not change during development but have a differing susceptibility according to age.

Animals↗
PubMed1997

Virus elimination and histologic improvement in patients with chronic hepatitis C treated with interferon alpha.

We evaluated the elimination of hepatitis C virus (HCV) by means of interferon alpha (IFN-alpha) by investigating both positive- and negative-stranded RNA forms in the peripheral blood mononuclear cells and liver tissue. We also assessed the long-term histologic improvement accompanying viral clearance. We studied 20 persons with HCV whose serum aminotransferase levels remained normal for more than 1.5 years after IFN-alpha treatment withdrawal. The presence of HCV RNA in their peripheral blood and of both positive and negative strands in the peripheral blood mononuclear cells and liver tissue was investigated using the reverse transcription polymerase chain reaction method. We examined the histologic findings using the European classification and the histology activity index scoring system. In 16 of 20 patients, both strands disappeared from the possible reservoirs. The histologic findings indicated reduced activity, and histology activity index scores (1, 2, 3, and total) also showed significant improvement. We confirmed that IFN-alpha therapy can induce the elimination of HCV RNA from the conceivable HCV reservoirs and effect histologic improvement. Therapy with IFN-alpha is effective for treating chronic hepatitis C.

Adult↗
PubMed1997

Antiperoxidative activity of neolignans from Magnolia obovata.

Honokiol and magnolol, neolignans in Magnolia obovata, have been evaluated as antioxidants. Microsomal lipid peroxidation induced by Fe(III)-ADP/NADPH and mitochondrial lipid peroxidation induced by Fe(III)-ADP/NADH were inhibited by these compounds. These neolignans protected mitochondrial respiratory chain enzyme activity against NADPH-induced peroxidative stress and protected red cells against oxidative haemolysis. The anti-oxidative activity of honokiol was more potent than that of magnolol. Neolignans in M. obovata were shown to be effective in protecting biological systems and functions against oxidative stress.

Animals↗
PubMed1997

Vulnerability to cerebral hypoxic-ischemic insult in neonatal but not in adult rats is in parallel with disruption of the blood-brain barrier.

BACKGROUND AND PURPOSE: Vulnerability to cerebral hypoxic-ischemic (H-I) insult and its relation to disruption of the blood-brain barrier were investigated in postnatal rats. METHODS: Pups of postnatal day (P) 7, P14, and P21 underwent ligation of a unilateral carotid artery and were exposed to hypoxic conditions. For the detection of early-phase deterioration, brains were perfusion-fixed 24 hours after H-I insult and examined by argyrophil III method. For the detection of later infarction, animals were fixed at 72 hours after the H-I insult. RESULTS: In either case, tissue damage was detected in the striatum, parietal cortex, and hippocampus. The vulnerability of P7 and P21 rats was remarkable, as compared with P14 rats. Although the developmental status of the vasculature was not significantly different at each age, the permeability of IgG after H-I injury was prominent in P7 rats and to a lesser extent in P14 rats. In P21 rats, however, there was little IgG leakage even 24 hours after the insult. Dexamethasone pretreatment blocked the extravasation of IgG and reduced the damaged tissue in P7 and P14 rats but not in P21 rats. Percentages of reduction in infarcted areas by the dexamethasone became smaller in proportion to ages. CONCLUSIONS: The results suggest that in younger rats vulnerability to H-I insult was in parallel with permeability of the blood-brain barrier, whereas in adults in might be more dependent on cellular vulnerability.

Aging↗
PubMed1997

Inhibition of aldose reductase and sorbitol accumulation by astilbin and taxifolin dihydroflavonols in Engelhardtia chrysolepis.

Dihydroflavonol taxifolin and its glycoside, astilbin, from Engelhardtia chrysolepis inhibited rat lens and recombinant human aldose reductase. Taxifolin also inhibited sorbitol accumulation in human red blood cells. Furthermore, this dihydroflavonol aglycone maintained the clarity of rat lens incubated with a high concentration of glucose. These dihydroflavonols may be effective for preventing osmotic stress in hyperglycemia.

Aldehyde Reductase↗
PubMed1997

[Diagnosis of tumor extension of ductal adenocarcinoma of the pancreas based on histological findings].

This study was undertaken to evaluate the diagnosis for tumor extension of ductal adenocarcinoma of the head of the pancreas based on histological investigation. From 1968 to 1995, 316 patients underwent radical resection and histological tumor extension was quite as follows; the rate of invasion to the anterior pancreatic capsule was 49%, invasion to the retroperitoneal tissue was 77%, invasion to the portal vein system was 38%, invasion to the extrapancreatic nerve plexus was 53% and lymph node metastases were 79%. 249 patients was performed extended radical operation consisted of regional lymphadenectomy, retroperitoneal dissection and resection of portal vein system, however non-curative resection was 52% with tumor invasion to dissected pancreatic surface in 88% of non-curative patients. And there was no 5-year survivor with non-curative resection. The extended radical operation should be indicated for patients who will have curative resection. Then, we set up clinical stage (CS; CSI approximately IV) by three factors related to resectability mostly; invasion to retroperitoneal tissue (RP), invasion to portal vein system (PV), invasion to major arterial system (A). It was also set up preoperative diagnostic criteria for RP, PV and A factor by computed tomography (CT) or abdominal angiography. From 1989 to 1995, 101 patients who had extended radical operation were investigated prospectively. The rate of accuracy of preoperative diagnosis of tumor extension were about 80% in each factor. Curability was 94% in CS I, 67% in CS II, 43% in CS III, respectively, and 3-year survival rate was 53% in CS I, 35% in CS II, 2-year survival rate was 8% in CS III. However, in CS IV the rate of non-curative resection was 77% and there was no 2-year survivor. It was concluded that extended radical operation of ductal adenocarcinoma of the head of the pancreas should be indicated for patients in less than CS III diagnosed by CT or angiography preoperatively.

Carcinoma, Ductal, Breast↗
PubMed1996

4-Hydroxy-2-nonenal cytotoxicity in renal proximal tubular cells: protein modification and redox alteration.

4-Hydroxy-2-nonenal (HNE), one of the major products of membrane lipid peroxidation, has been shown recently to be present in a form covalently attached to proteins in the renal proximal tubules of rats treated with a renal carcinogen, ferric nitrilotriacetate (Toyokuni, S., et al. (1994) Proc. Natl. Acad. Sci. USA 91, 2616-2620; Uchida, K., et al. (1995) Arch. Biochem. Biophys. 317, 405-411). In the present study, the mechanism of HNE cytotoxicity was studied using the renal tubular epithelial cells (LLC-PK1), focusing on the protein modification and alteration of cellular redox status induced by HNE. Upon treatment with HNE for 2 h, the LLC-PK1 cells were found to be resistant to the low concentration (10 microM) of HNE, while HNE at higher concentrations (> or = 50 microM) mediated cell death. The cytotoxicity of HNE appeared to be correlated with the HNE modification of cellular proteins. Among a number of proteins modified by HNE, a glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase was detected as one of the major targets of HNE in the cells. On the other hand, exposure of LLC-PK1 cells to HNE resulted in rapid reduction of cellular glutathione (GSH) levels, suggesting that HNE influenced primarily the redox status of the cells. Depletion of GSH with buthionine sulfoximine, a potent suppressor of GSH biosynthesis, before HNE treatment caused the cells to be sensitive to HNE cytotoxicity and to HNE modification of cellular proteins, whereas the increase in intracellular GSH levels by treatment with N-acetylcysteine before HNE treatment resulted in a dose-dependent inhibition of HNE-mediated protein modification. These results suggest that intracellular GSH is a determinant on cellular resistance against the HNE-mediated cytotoxicity.

Acetylcysteine↗
PubMed1996

Oxidative stress response in iron-induced renal carcinogenesis: acute nephrotoxicity mediates the enhanced expression of glutathione S-transferase Yp isozyme.

An iron chelate, ferric nitrilotriacetate (Fe-NTA), induces acute renal proximal tubular necrosis, a consequence of free radical-mediated oxidative tissue damage, that eventually leads to a high incidence of renal adenocarcinoma in rodents. In the present study, we investigated the free radical-induced oxidative stress response in this carcinogenesis model, focusing on the expression of glutathione S-transferases (GSTs) which catalyze the conjugation of reactive chemicals with glutathione and play an important role in protecting cells. A single intraperitoneal Fe-NTA treatment (15 mg Fe/kg body weight) induced a rapid oxidative stress, which was monitored by the accumulation of lipid peroxidation products and the loss of sulfhydryl contents in the kidneys, resulting in a 30% reduction of GST activity 1 h after an Fe-NTA treatment. The enzyme activity returned to the control level after 16 h. The immunoblot analysis of GST isozymes demonstrated that the level of alpha-class GSTs (GST-Ya and GST-Yc) and pi-class GST (GST-Yp), major GST isozymes constitutively produced in the kidney, decreased immediately within 1 h of the Fe-NTA treatment. The onset of the recovery of GST-Yp protein levels was detected 3 h after the Fe-NTA treatment. The enhanced production of GST-Yp in gene expression was evident in the drastic elevation of mRNA levels and these increases coincided with a substantial rise in the GST activity and protein levels. The alpha-class GSTs were not inducible by treatment with Fe-NTA. The immunohistochemical analysis demonstrated that the expression of GST-Yp was strongly induced in the regenerating proximal tubular cells. A steady accumulation of GST-Yp protein was observed in the subacute toxicity experiments with multiple injections of Fe-NTA. These results suggest that the enhanced expression of GST-Yp is important in mediating cell repairs or increasing the resistance to subsequent injury.

Adenocarcinoma↗
PubMed1996

Tissue extract from dopamine-depleted striatum enhances differentiation of cultured striatal type-1 astrocytes.

The effects of tissue extract from dopamine (DA)-depleted striatum (lesion extract, L-ext) on morphological and electrophysiological natures of cultured striatal astrocytes were investigated. L-ext treatment suppressed the proliferation of type-1 astrocytes. They became fibrous in a concentration-dependent manner. These changes were not observed in type-2 astrocytes. By whole cell patch-clamp recording, two kinetically and pharmacologically distinct voltage-activated potassium currents, A current and delayed rectifier, were identified. L-ext treatment enhanced both currents in type-1 astrocytes, but only A current in type-2. Data suggest that in tissue extract from DA-depleted striatum, there are increased trophic activities that promote the differentiation of type-1 astrocytes.

Animals↗
PubMed1996

Oxidative stress response in iron-induced acute nephrotoxicity: enhanced expression of heat shock protein 90.

Iron overload with ferric nitrilotriacetate (Fe-NTA) induces acute renal proximal tubular necrosis, a consequence of oxidative tissue damage, that leads to a high incidence of renal adenocarcinoma in rodents. In the present study, we determined the proteins preferentially produced in response to the Fe-NTA-induced oxidative injury. A single intraperitoneal Fe-NTA treatment led to the enhanced production of a number of proteins with molecular masses of 85-95 kDa. These included heat shock protein 90 (HSP90) as determined by immunoprecipitation. The enhanced production of HSP90 was prominent in the renal tubular cells. Steady accumulation of HSP90 was observed in the subacute toxicity experiments with multiple injections of Fe-NTA, suggesting that the enhanced production of HSP90 is important in increasing resistance to subsequent injury caused by the Fe-NTA-induced oxidative stress.

Animals↗
PubMed1996

Labeling and identification of living donor cells in brain slices of recipient hemiparkinsonian model rats for physiological recordings: methods for physiological assessments of neural transplantation.

Physiological properties of grafted neurons, such as membrane and intr acellular properties, have not been reported. To fill this lack in knowledge, physiological recordings from the identified grafted cells are required. Fluorescent latex microspheres (FLM) are non-toxic and stable, and thus seem suitable for long-term labeling of donor cells. Therefore, we tested the feasibility of labeling with FLM to identify living donor cells in the recipients' brain slices. We also tested if physiological recordings from the identified cells are possible or not. Cell suspensions were prepared from the substantia nigra (SN) of Embryonic Days 15 or 16 rats with enzymatic and mechanical trituration. Cell suspensions were then incubated with 0.5% FLM for 30 min to 2 h. The longer cells were incubated, the more FLM were taken up. The FLM-labeled SN cell suspensions were injected in the striatum of the hemiparkinsonian model rats. Eight to 13 weeks later, 150-mu m thick coronal brain slices including the graft track were prepared from the recipients. Slices were kept in vitro for several hours. Grafted cells could be clearly identified in the slice preparations by the uptake of FLM under a fluorescence microscope. Voltage-dependent currents and intracellular Ca2+ transcients were successfully recorded from the identified grafted neurons. It is suggested that labeling and identification of living donor cells with FLM is feasible and thus can provide a powerful tool to study the mechanisms underlying graft-induced amelioration of neurological deficits in parkinsonism by enabling physiological assessments of grafted cells.

Animals↗
PubMed1996

Early cytopathic features in rat ischemia model and reconstruction by neural graft.

Using a silver impregnation (argyrophil III) and immunohistochemistry, acute cytopathic features after cerebral ischemia were investigated. Additionally, functional recovery and interconnection between the host and graft was also explored after neural graft. Animals were embolized in unilateral middle cerebral artery for 1 h. Argyrophil III method demonstrated "collapsed" dark neurons in the striatum, cortex, reticular thalamus, amygdala, and hypothalamus on ischemic side. These neurons exhibited characteristic shrunken somata with corkscrew-like dendrites, suggesting changes in cytoskeletal protein. In the above mentioned areas, the loss of immunoreactivity for mu-calpain proenzyme and microtubule-associated protein 2 was also detected. Neural graft into the ischemic striatum was made 2 weeks after the ischemia paradign. The grafted striatal cells were prepared from E15 fetuses to make cell suspension marked by rhodamine-labeled latex microspheres. Methamphetamine-evoked rotations were detected after ischemia. These motor alterations were reduced gradually but significantly at 8 weeks after the graft. Interconnecton between the host and grafted cells was then studied in a brain slice preparation after loading fura-2 AM. About 10% of grafted cells tested from rats that showed motor amelioration exhibited [Ca2+]i increase to the electrical stimulation applied to the neighboring host tissue. Data indicate that, in the very early stage after ischemia, cytoskeletal damages, especially on microtubules, started and this would lead to later infarct. The graft survived in the ischemic striatum having connections with the host, and this might be partly involved in the amelioration of motor function.

Animals↗
PubMed1996

Microtubule-associated protein 2 expressing COS7 cells are resistant to argyrophilia under oxygen- and glucose-free condition.

Argyrophil III silver impregnation is a very sensitive method to detec t the early damage to neurons following brain ischemia. The argyrophil III staining and microtubule-associated protein 2 (MAP2) immunocytochemistry were performed on PC12D cells and MAP2C cDNA-transfected COS7 cells to detect the changes cytoskeletal proteins (microtubules/MAP2). After exposing these cells to simulated ischemic condition (oxygen and/or glucose free), the correlation between the appearance of the argyrophilia and the disappearance of MAP2 was investigated. The PC12D cells expressed very low MAP2 and became argyrophilic very easily depending on the degree of the ischmeia-like insult, whereas MAP2C cDNA-transfected COS7 cells expressed a higher level of MAP2C and were resistant to argyrophilia, although the diameter of their immunoreactive processes became thinner. Thus, when MAP2C is expressed at a higher level, cells became resistant to argyrophilia, suggesting a correlation between the argyrophilia and the damage on microtubules and MAPs.

Animals↗
PubMed1996

Parameters for plaque formation in the potency assay of Japanese measles vaccines.

Parameters for plaque formation by measles vaccine strains licensed in Japan were studied. For the plaque test, inoculum volume was one of the critical factors for obtaining an appropriate titre of the sample. A linear relationship between the inoculum volume and the apparent reciprocal titre was discovered, enabling the comparison of absolute titres. Another factor affecting the infectivity was the strain-specific temperature sensitivity in the plaque assay. Although all the vaccine strains tested showed the highest titre at 35 degrees C, the pattern of the temperature sensitivity differed from one strain to another. These factors must be taken into consideration in order to obtain an appropriate titre of a vaccine virus.

Animals↗