Biomedical subjects

T Funakoshi

Publications and source records attributed to T Funakoshi.

At least 19 recordsLinked to original sources

PubMed2006

Extracellular matrix modulates expression of cell-surface proteoglycan genes in fibroblasts.

Cell-surface proteoglycans are involved in many functions, including interactions with components of the extracellular microenvironment. They also act as coreceptors that bind and modify the actions of various growth factors, cytokines, and the extracellular matrix (ECM). This study investigated the regulation by the ECM of the expression of cell-surface proteoglycans (CD44, syndecan-1-4, betaglycan, glypican-1). We examined the changes in the expression levels of cell-surface proteoglycan genes in intact tendon, monolayer culture, and under various culture conditions. There was a significant increase in the expression of CD44 and syndecan-4 mRNAs during cell isolation from the tendon. With the switch to a 3D culture environment, there was a significant increase in the expression of CD44 at each passage point relative to its expression in 2D at those passage points. Syndecan-4 mRNA also increased steadily at each passage point in 3D culture environment. This influence on cell surface proteoglycans gene expression may indicate that collagen gel culture mimics in vivo tendon environment. This study provides further insight into the regulation of cell-surface proteoglycans in ligament and tendon fibroblasts by the ECM and 3D culture conditions.

Animals↗

PubMed2005

Pregnancy complicated with pulmonary lymphangioleiomyomatosis: case report.

A case of a 30-year-old primiparous woman with pulmonary lymphangioleiomyomatosis is described. The patient had experienced six episodes of spontaneous pneumothorax at the age of 27 years and had been diagnosed as having pulmonary lymphangioleiomyomatosis based on histological findings of specimens obtained by transbronchial biopsy. She had undertaken open lung surgery and thoracoscopy. Thereafter, she became pregnant spontaneously. Her antenatal course was uneventful with no exacerbation of respiratory status. At 38 weeks of gestation, she underwent a selective cesarean section and myomectomy under combined spinal and epidural anesthesia. Her postoperative course was uneventful. No remarkable changes in computed tomographic findings of the lung were noted on the 20th day of postoperation compared with those before pregnancy. She has been followed-up in the pulmonary outpatient clinic with no deterioration of the disease.

Adult↗

PubMed2005

Consumptive coagulopathy that developed in a pregnant woman with degenerated uterine leiomyoma: case report.

A case of consumptive coagulopathy in a 30-year-old pregnant woman with degenerated uterine leiomyoma is described. She developed lower abdominal pain at 21 weeks of gestation. Laboratory profiles revealed coagulation abnormalities. Magnetic resonance imaging diagnosed degenerated uterine leiomyoma. At 34 weeks, she underwent cesarean section and myomectomy. Pathological examination showed the presence of organized thrombi in the vessels of leiomyoma. The consumption of platelets and coagulation factors in degenerated uterine leiomyoma may result in consumptive coagulopathy.

Adult↗

PubMed2005

A pregnant woman with anti-Gregory antigen: case report.

Negative Gregory antigen (Gy(a-)) remains an extremely uncommon blood phenotype. We describe a 32-year-old pregnant woman with (Gy(a-)) and anti-Gregory antigen (anti-Gy(a)). There was no evidence of consanguineous mating in her family. Blood typing study revealed that only her father was Gy(a-) among the family. Anti-Gy(a) had a titer of 16 before pregnancy, but increased to 1024 at 33 weeks of gestation with a titer of 512 at 34 weeks. Her own blood stores were collected starting at 14 weeks, amounting to 1800 g totally. She underwent an emergency cesarean section at 35 weeks due to a non-reassuring fetal status. Blood loss was approximately 1090 g. Cord blood type was found to be Gy(a-). The indirect Coombs test of cord blood was positive, while the direct Coombs test was negative. No neonatal hemorrhagic disease developed. The storage of a sufficient amount of crossmatch-compatible Gy(a-) blood during pregnancy is important in case of possible need of blood transfusion at delivery for women with anti-Gy(a).

Adult↗

PubMed2004

Effect of re-expansion after short-period lung collapse on pulmonary capillary permeability and pro-inflammatory cytokine gene expression in isolated rabbit lungs.

BACKGROUND: Re-expansion pulmonary oedema is a rare complication caused by rapid re-expansion of a chronically collapsed lung. Several cases of pulmonary oedema associated with one-lung ventilation (OLV) have been reported recently. Elevated levels of pro-inflammatory cytokines in pulmonary oedema fluid are suggested to play important roles in its development. Activation of cytokines after re-expansion of collapsed lung during OLV has not been thoroughly investigated. Here we investigated the effects of re-expansion of the collapsed lung on pulmonary oedema formation and pro-inflammatory cytokine expression. METHODS: Lungs isolated from female white Japanese rabbits were perfused and divided into a basal (BAS) group (n=7, baseline measurement alone), a control (CONT) group (n=9, ventilated without lung collapse for 120 min) and an atelectasis (ATEL) group (n=9, lung collapsed for 55 min followed by re-expansion and ventilation for 65 min). Pulmonary vascular resistance (PVR) and the coefficient of filtration (Kfc) were measured at baseline and 60 and 120 min. At the end of perfusion, bronchoalveolar lavage fluid/plasma protein ratio (B/P), wet/dry lung weight ratio (W/D) and mRNA expressions of tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta and myeloperoxidase (MPO) were determined. RESULTS: TNF-alpha and IL-1beta mRNA were significantly up-regulated in lungs of the ATEL group compared with BAS and CONT, though no significant differences were noted in PVR, Kfc, B/P and W/D within and between groups. MPO increased at 120 min in CONT and ATEL groups. CONCLUSION: Pro-inflammatory cytokines were up-regulated upon re-expansion and ventilation after short-period lung collapse, though no changes were noted in pulmonary capillary permeability.

Animals↗

PubMed2004

[Aberrant subclavian artery].

Aberrant subclavian artery (ASA) is the most frequently encountered congenital anomaly of aortic arch. The ASA aneurysms are rare but potentially lethal disease, it has been recognized that the presence of an aneurysm of an ASA itself is an indication for surgery. We experienced 4 cases with ASA during the past 23 years. All cases were male. Their ages ranged from 51 to 73 years old. Three patients had a right-sided aortic arch. Among the 4 patients, only 1 underwent total arch replacement, but the other 3 patients could be followed conservatively for 1, 7 and 23 years, respectively. We reviewed the problem of the diagnosis and treatment of the ASA including surgical indication.

Aged↗

PubMed2001

Gene therapy utilizing the Cre/loxP system selectively suppresses tumor growth of disseminated carcinoembryonic antigen-producing cancer cells.

Recent clinical trials of cancer gene therapy have shown encouraging results for controlling localized tumors. However, to control metastatic or disseminated tumor cells, further modification of vectors is required to enhance specificity and infectivity against targets. We investigated whether utilization of the Cre recombinase(Cre)/loxP system contributes to enhanced antitumor effects together with minimal adverse reactions in specific gene therapy against disseminated carcinoembryonic antigen (CEA)-producing cancer cells in the peritoneal cavity of mice. CEA-producing cancer would be a good therapeutic target because it is found in lung, stomach and colon sites, which account for most cancers. We constructed a pair of recombinant adenoviral vectors (Ads), one of which expresses the Cre gene under the control of the CEA promoter (Ad.CEA-Cre); the other expresses the herpes simplex virus thymidine kinase (HSV-TK) gene (Ad.lox-TK), or the beta-galactosidase gene (beta-gal) by Cre (Ad.lox-beta-gal). Intraperitoneal coinjection of Ad.CEA-Cre and Ad.lox-beta-gal into mice with peritonitis carcinomatosa by CEA-producing tumor cells showed selective expression of the beta-gal gene in tumor foci. Coinfection of Ad.CEA-Cre and Ad.lox-TK followed by ganciclovir (GCV) administration significantly suppressed the total tumor weight in the peritoneal cavity of the mice to 13% of that of the untreated mice and 22% of that of the mice treated with Ad.CEA-TK/GCV, an Ad that expressed the HSV-TK gene driven by the CEA promoter alone. Moreover, treatment with Ad.CEA-Cre and Ad.lox-TK/GCV completely suppressed tumors in 4 of 10 (40%) mice without significant weight loss, although 2 of 10 mice treated with Ad.CAG-TK/GCV, an adenovirus vector that strongly but nonspecifically expressed the TK gene, died due to severe side effects including diarrhea, weight loss and liver dysfunction. These findings suggest that cell type-specific gene therapy using the Cre/loxP system is effective against disseminated cancer cells without significant side effects.

Adenoviridae↗

PubMed2001

Neuropharmacological profiles of a novel atypical antipsychotic, NRA0562, in rats.

Neuropharmacological profiles of 5-(2-[4-(6-fluoro-1H-indole-3-yl) piperidine-1-yl] ethyl)-4-(4-fluorophenyl) thiazole-2-carboxylic acid amide (NRA0562) in rats were examined using electrophysiological and immunohistochemical methods. The firing rates of the substantia nigra pars compacta (A9) and the ventral tegmental area (A10) dopamine neurons were inhibited by methamphetamine (1 mg/kg, i.v.). NRA0562 dose-dependently reversed the inhibitory effects of methamphetamine on A9 and on A10 dopamine neurons. NRA0562 was more potent to reverse the inhibitory effects of methamphetamine on A10 (ED(50)=0.3 mg/kg) than on A9 (ED(50)=0.9 mg/kg) dopamine neurons. NRA0562 produced significant increases in Fos-like immunoreactivity in both the nucleus accumbens and the dorsolateral striatum. The difference between the number of Fos-like immunoreactivity produced by NRA0562 in the nucleus accumbens vs. dorsolateral striatum, referred to as the atypical index, was positive. Similar results could be observed with risperidone, an atypical antipsychotic. These results suggest that NRA0562 may have the atypical antipsychotic activities seen with risperidone, but without the liability of motor side effects typical of classical antipsychotics.

Action Potentials↗

PubMed2001

Adenovirus-mediated gene therapy specific for small cell lung cancer cells using a Myc-Max binding motif.

Recent clinical trials of gene therapy for patients with thoracic cancers have shown that these treatments were well tolerated with minimal side effects and that we need to further enhance specificity as well as efficiency of gene transfer to target cancer cells. We previously reported that myc-overexpressing SCLC cell lines became selectively sensitive to ganciclovir (GCV) by transducing the herpes simplex virus thymidine kinase (HSV-TK) gene under the control of the Myc-Max response elements (a core nucleotide sequence, CACGTG) and that this construct (MycTK) could be utilized to develop a novel treatment against chemo-radio-resistant SCLC. We report here in vivo antitumor effects and safety of a replication-deficient adenoviral vector containing the Myc-Max binding motif (AdMycTK) on SCLC cells. In vitro infection with AdMycTK selectively rendered myc-overexpressing SCLC cell lines 63- to 307-fold more sensitive to GCV. In vivo injections with AdMycTK followed by GCV administration markedly suppressed the growth of myc-overexpressing tumors established in the subcutis or in the peritoneal cavity of athymic mice. On the other hand, infection with AdMycTK did not significantly affect either in vitro GCV sensitivity of the cells expressing very low levels of the myc genes or the growth of their subcutaneous tumors. Moreover, we observed no apparent side effects of this treatment including body weight loss or biochemical abnormalities in contrast to the treatment with AdCATK that conferred strong but nonspecific expression of the HSV-TK gene. These results suggested that AdMycTK/GCV therapy is effective on SCLC patients whose tumors overexpress myc family oncogenes.

Adenoviridae↗

PubMed2001

Interaction of tyrosine 65 of RecA protein with the first and second DNA strands.

We investigated the structure of the active RecA-DNA complex by analyzing the environment of tyrosine residue 65, which is on the DNA-binding surface of the protein. We prepared a modified RecA protein in which the tyrosine residue was replaced by tryptophan, a natural fluorescent reporter, and measured the change in its fluorescence upon binding of DNA and cofactor. The fluorescence of the inserted tryptophan 65 (Trp65) was centered at 345 nm, indicating a partly exposed residue. Binding cofactor, adenosine 5'-O-3-thiotriphosphate (ATPgammaS), alone at a low salt concentration did not change the fluorescence of Trp65, confirming that the residue is not close to the nucleotide. In contrast, the binding of single-stranded DNA quenched the fluorescence of Trp65 in both the presence and absence of ATPgammaS. Trp65 fluorescence was also quenched upon binding a second DNA strand. The fluorescence change depended upon the presence and absence of ATPgammaS, reflecting the difference in the DNA binding. These results indicate that residue 65 is close to both the first and second DNA strands. The degree of quenching depended upon the base composition of DNA, suggesting that the residue 65 interacts with the DNA bases. Binding of DNA with ATPgammaS as well as binding of ATPgammaS alone at high salt concentration shifted the fluorescence emission peak from 345 to 330 nm, indicating a change from a polar to a non-polar environment. Therefore, the environment change around residue 65 would also be linked to a change in conformation and thus the activation of the protein.

Adenosine Triphosphatases↗

PubMed2001

Differences in incision shape based on the keratome bevel.

PURPOSE: To evaluate differences in incision shape due to the bevel of the keratome. SETTING: Kushimoto Rehabilitation Center, Kushimoto, Japan. METHODS: Three types of keratomes (bibevel, bevel up, and bevel down) were inserted into soft polyvinyl chloride sheets and polyurethane sheets, and the incision shapes in the blade entrance and exit planes were evaluated. The experiments were repeated using soft polyvinyl chloride domes and silicone domes. RESULTS: In the experiments using artificial sheets, the incisions in the blade entrance and exit planes using the bibevel keratome had a linear shape, those using the bevel-up keratome had an inverse-V shape, and those using the bevel-down keratome had a V shape. In the experiments using artificial domes, the incision shapes were the same as those in the artificial sheets. CONCLUSIONS: The incision shape differed according to the direction of the keratome bevel. These experiments and results may provide useful data when new knives are developed or when incision shapes following intended specifications are stably made.

Cornea↗

PubMed2001

Selection of eligible patients with supratentorial glioblastoma multiforme for gross total resection.

The purpose of this study is to clarify whether gross total tumor resection can prolong the survival in adult patients with supratentorial glioblastoma multiforme (GBM), and to clarify what subset of these patients obtains a survival advantage by gross total tumor resection without postoperative neurological deterioration. Eighty-two adult patients with supratentorial GBM were retrospectively reviewed. Overall, the median survival time was 13 months, and the 1- and 2-year survival rates were 53.7% and 14.6%, respectively. In a univariate analysis for survival rate by log-rank test, age (< 40 years), Karnofsky performance scale (KPS) score (70-100%) and extent of surgery (gross total resection) were revealed to be significant good prognostic factors. A Cox proportional hazard multivariate regression analysis confirmed that the KPS and extent of surgery were independent, significant good prognostic factors. Nine patients (11%) suffered postoperative neurological deterioration. A topographical GBM staging system (Stages I, II and III) with the integration of tumor location, size and eloquence of adjacent brain based on MRI (for explanation of Stages see text) was originally proposed. In Stage I, gross total resection had a strong tendency toward a better prognostic factor in a univariate analysis and was revealed to be a significant independent good prognostic factor in a multivariate analysis. In also Stage II, the survival of patients who underwent gross total resection was better than that of patients with less than gross total resection, although not significant. In Stage III, there were no patients who underwent gross total tumor resection. Risk probabilities of postoperative neurological deterioration, overall, were 0%, 22.2%, and 20% in Stages I, II, and III, respectively, and those after gross total resection were 0% and 16.7% in Stages I and II, respectively. Although gross total tumor resection is associated with prolongation of the survival time of patients with GBM, the risk of postoperative neurological deficit increases with radical tumor resection. To select an eligible subset of patients that benefit in survival from gross total tumor resection without postoperative risk, the following surgical policy for GBM resection is suggested. GBM in Stage I should be resected as radically as possible. Regarding Stage II, risky surgical resection extending to the area adjacent to the critical zone should be avoided and more meticulous and careful surgical planning is needed than that in Stage I. In Stage III, radical gross total tumor resection is not recommended at present.

Adult↗

PubMed2001

Pur alpha protein implicated in dendritic RNA transport interacts with ribosomes in neuronal cytoplasm.

We have previously reported that pur alpha, known to be a regulator of DNA replication and transcription, links neural BC1 RNA to microtubules via dendrite-targeting RNA motifs. Here we demonstrate the subcellular localization of pur proteins within the brain. Pur proteins were detected in neurons but not in glia. Immunohistochemical staining was prominent in perikarya and proximal dendrites and also extended into primary dendritic processes, but no significant signals were detected in the distal regions of dendrite. When homogenates of mouse brain were fractionated, pur alpha was most concentrated in the microsomal pellet. Consistently, pur alpha co-fractionated with free polysomes as well as with membrane-bound polysomes and the association with polysomes was mediated by binding ribosomal subunits. Levels of ribosomes with pur alpha progressively increased during postnatal development of the brain.

Animals↗

PubMed2000

The effects of sulfhydryl blockers and metal ions on nickel accumulation by rat primary hepatocyte cultures.

Previously, we found that nickel (Ni) accumulation by rat hepatocytes involves Ca channel transport processes. However, other mechanisms responsible for Ni accumulation are still unclear. Therefore, in the present study we examined the effects of sulfhydryl (SH) blockers on Ni accumulation by hepatocytes. Hepatocytes were exposed to various concentrations of N-ethylmaleimide (NEM) (0.5, 1 or 2 mM) or monoiodoacetic acid (MIA) (0.1, 0.25 or 0.5 mM), potent blockers of SH ligands, for 30 min and subsequently exposed to 10 microM NiCl(2) for an additional 60 min. Pretreatment with NEM and MIA enhanced the Ni accumulation by hepatocytes to maximum of 156 and 73%, respectively. The effects of essential and nonessential metal ions on Ni accumulation were also investigated. Pretreatment with 10 microM of Cu, Zn, Co, Cd and Mn, decreased Ni accumulation by 46, 30, 20, 18 and 11%, respectively. In contrast, pretreatment with Hg (10 and 20 microM) enhanced the Ni accumulation by almost 81 and 140%, respectively. Furthermore, significant decrease in SH concentration in the hepatocyte membrane was observed by the treatment with NEM, MIA and Hg, but not with Cu, Zn and Cd. These results suggest that Ni accumulation by hepatocytes does not appear to be dependent on the SH carrier-mediated transport processes, and that to block the SH ligands in the plasma membrane may facilitate the Ni crossing of the cell membrane.

Animals↗

PubMed2000

Reconstitution of ATP- and cytosol-dependent transport of de novo synthesized ceramide to the site of sphingomyelin synthesis in semi-intact cells.

Transport of ceramide synthesized at the endoplasmic reticulum to the Golgi compartment, where sphingomyelin (SM) synthase exists, was reconstituted within semi-intact Chinese hamster ovary cells. When [(3)H]ceramide that had been produced from [(3)H]sphingosine at 15 degrees C in perforated cells was chased at 37 degrees C, [(3)H]ceramide-to-[(3)H]SM conversion occurred in a cytosol-dependent manner. In various aspects (i.e. kinetics, ATP dependence, and temperature dependence), [(3)H]ceramide-to-[(3)H]SM conversion in perforated cells was consistent with that in intact cells. The cytosol from LY-A strain, a Chinese hamster ovary cell mutant defective in endoplasmic reticulum-to-Golgi transport of ceramide, did not support [(3)H]ceramide-to-[(3)H]SM conversion in perforated wild-type cells, whereas the wild-type cytosol rescued the conversion in perforated LY-A cells. Brefeldin A-treated cells, in which the endoplasmic reticulum and the Golgi apparatus were merged, no longer required cytosol for conversion of [(3)H]ceramide to [(3)H]SM. These results indicated that the assay of [(3)H]ceramide-to-[(3)H]SM conversion in semi-intact cells is a faithful in vitro assay for the activity of cytosol-dependent transport of ceramide and that LY-A cells are defective in a cytosolic factor involved in ceramide transport. In addition, conversion of [(3)H]ceramide to [(3)H]glucosylceramide in semi-intact cells was little dependent on cytosol, suggesting that ceramide reached the site of glucosylceramide synthesis by a cytosol-independent (or less dependent) pathway.

Adenosine Triphosphate↗

PubMed2000

Tor-mediated induction of autophagy via an Apg1 protein kinase complex.

Autophagy is a membrane trafficking to vacuole/lysosome induced by nutrient starvation. In Saccharomyces cerevisiae, Tor protein, a phosphatidylinositol kinase-related kinase, is involved in the repression of autophagy induction by a largely unknown mechanism. Here, we show that the protein kinase activity of Apg1 is enhanced by starvation or rapamycin treatment. In addition, we have also found that Apg13, which binds to and activates Apg1, is hyperphosphorylated in a Tor-dependent manner, reducing its affinity to Apg1. This Apg1-Apg13 association is required for autophagy, but not for the cytoplasm-to-vacuole targeting (Cvt) pathway, another vesicular transport mechanism in which factors essential for autophagy (Apg proteins) are also employed under vegetative growth conditions. Finally, other Apg1-associating proteins, such as Apg17 and Cvt9, are shown to function specifically in autophagy or the Cvt pathway, respectively, suggesting that the Apg1 complex plays an important role in switching between two distinct vesicular transport systems in a nutrient-dependent manner.

Adaptor Proteins, Signal Transducing↗

PubMed2000

Apg13p and Vac8p are part of a complex of phosphoproteins that are required for cytoplasm to vacuole targeting.

We have been studying protein components that function in the cytoplasm to vacuole targeting (Cvt) pathway and the overlapping process of macroautophagy. The Vac8 and Apg13 proteins are required for the import of aminopeptidase I (API) through the Cvt pathway. We have identified a protein-protein interaction between Vac8p and Apg13p by both two-hybrid and co-immunoprecipitation analysis. Subcellular fractionation of API indicates that Vac8p and Apg13p are involved in the vesicle formation step of the Cvt pathway. Kinetic analysis of the Cvt pathway and autophagy indicates that, although Vac8p is essential for Cvt transport, it is less important for autophagy. In vivo phosphorylation experiments demonstrate that both Vac8p and Apg13p are phosphorylated proteins, and Apg13p phosphorylation is regulated by changing nutrient conditions. Although Apg13p interacts with the serine/threonine kinase Apg1p, this protein is not required for phosphorylation of either Vac8p or Apg13p. Subcellular fractionation experiments indicate that Apg13p and a fraction of Apg1p are membrane-associated. Vac8p and Apg13p may be part of a larger protein complex that includes Apg1p and additional interacting proteins. Together, these components may form a protein complex that regulates the conversion between Cvt transport and autophagy in response to changing nutrient conditions.

Adaptor Proteins, Signal Transducing↗

PubMed2000

Primary primitive neuroectodermal tumor of the lung: report of two cases.

Two cases of primitive neuroectodermal tumor of the lung are reported. The first case is a 41-year-old man with a tumor in the left upper lung, and the second case is a 30-year-old woman with a tumor in the right lower lung. In both cases, the tumors originated in the lung but not in the chest wall. No distant metastasis was detected. In case 1, transcutaneous fine-needle biopsy (TCNB) revealed small round cell proliferation, although bronchoscopic examination showed no abnormal findings. Both the expression of Mic2 protein and t(11;22)(q24;q12) translocation were proved in the tumor cells. The tumor cells were positive for periodic acid-Schiff (PAS), neuron-specific enolase (NSE), and vimentin, but negative for Leu7, chromogranin A, and pro-gastrin-releasing peptide (ProGRP). In case 2, bronchoscopic examination showed only compressive change in right lower lobe bronchi. TCNB revealed small round tumor cells expressing Mic2 protein. The tumor cells were negative for leukocyte common antigen, S100 protein, pankeratin, chromogranin A, and desmin, but weakly positive for NSE and moderately positive for Ki-67 (MIB1). Both patients were successfully treated by the combination of surgical resection and chemotherapy, and are alive with no sign of recurrence for approximately 22 months in case 1 and 16 months in case 2.

12E7 Antigen↗