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Biomedical subjects

T Hirata

Publications and source records attributed to T Hirata.

At least 91 records · Page 5Linked to original sources

A cytoplasmic RNA vector derived from nontransmissible Sendai virus with efficient gene transfer and expression.

We have recovered a virion from defective cDNA of Sendai virus (SeV) that is capable of self-replication but incapable of transmissible-virion production. This virion delivers and expresses foreign genes in infected cells, and this is the first report of a gene expression vector derived from a defective viral genome of the Paramyxoviridae. First, functional ribonucleoprotein complexes (RNPs) were recovered from SeV cloned cDNA defective in the F (envelope fusion protein) gene, in the presence of plasmids expressing nucleocapsid protein and viral RNA polymerase. Then the RNPs were transfected to the cells inducibly expressing F protein. Virion-like particles thus obtained had a titer of 0.5 x 10(8) to 1. 0 x 10(8) cell infectious units/ml and contained F-defective RNA genome. This defective vector amplified specifically in an F-expressing packaging cell line in a trypsin-dependent manner but did not spread to F-nonexpressing cells. This vector infected and expressed an enhanced green fluorescent protein reporter gene in various types of animal and human cells, including nondividing cells, with high efficiency. These results suggest that this vector has great potential for use in human gene therapy and vaccine delivery systems.

Animals↗

Effects of dibutyryl cyclic adenosine monophosphate on the ultrastructure of endothelial cells in rat lungs cold preserved for 15 hours.

BACKGROUND: Dibutyryl cyclic adenosine monophosphate (db-cAMP) has been shown to protect vascular endothelial cells by increasing the level of intracellular cAMP, and we have previously reported its effectiveness in lung preservation. Here, the effects of db-cAMP in lung preservation were ultrastructurally investigated, and the ultrastructural changes before reperfusion were correlated with pulmonary function after reperfusion. METHODS: The lungs of 17 Lewis rats were flushed with perfusate and prostaglandin E(1), and were then divided into three groups. In the fresh group (n = 6), the lungs were flushed with extracellular-type trehalose-containing (ET-K) solution and were reperfused immediately. In the control group (n = 6) and db-cAMP group (n = 5), the lungs were flushed with ET-K solution and ET-K solution plus db-cAMP (2 mM), respectively, and were reperfused after cold preservation at 4 degrees C for 15 h. Before reperfusion, tissue was sampled and ultrastructurally analyzed by transmission electron microscopy. RESULTS: In the endothelial cells of pulmonary arterioles, the incidence of protrusion was significantly lower in the fresh and db-cAMP groups than in the control group (p < 0.05). The incidence of detachment and microvillus formation were significantly lower in the fresh and db-cAMP groups than in the control group (p < 0.01). The ultrastructure of the alveoli did not allow separation of the control and db-cAMP groups. The shunt fraction and wet to dry weight ratio of the lung tissue after reperfusion were significantly lower in the fresh and db-cAMP groups than in the control group (p < 0.01). Positive correlations were found between the incidence of these ultrastructural changes in the endothelial cells of the pulmonary arterioles and pulmonary function after reperfusion. CONCLUSION: These findings suggest that db-cAMP might attenuate the lung injury caused by cold preservation and ischemia-reperfusion, partly by suppressing the acceleration of the structural changes in the endothelial cells in the pulmonary arterioles.

Animals↗

Isolation and sequencing of a cDNA clone encoding a 20-kDa protein with trypsin inhibitory activity.

The amino acid sequence of a novel trypsin inhibitor (p20) was completed by the molecular cloning of the protein in cultured soybean cells. The clone nucleotide contains an open reading frame encoding a polypeptide of 206 amino acids that shows 45-50% sequence homology to members of the Kunitz-type trypsin inhibitor family. The p20 transcript is expressed in the roots, stems and leaves of soybean seedlings. DNA gel blot analyses show that the p20 in soybean is encoded by a single gene, and that this gene may not contain an intron.

Amino Acid Sequence↗

Inhibition of tumor cell growth by a specific 6-phosphofructo-2-kinase inhibitor, N-bromoacetylethanolamine phosphate, and its analogues.

The high rate of glycolysis despite the presence of oxygen and mitochondria in tumor cells implies an important role for this process in cell division. The rate of glycolysis is assumed to be dependent on the cellular concentration of fructose 2,6-bisphosphate, the concentration of which in turn depends on a bifunctional enzyme and the ratio of this enzyme's 6-phosphofructo-2-kinase versus its fructose 2,6-bisphosphatase activities. To prove the hypothesis that inhibition of glycolysis in tumor cells by 6-phosphofructo-2-kinase inhibitors would cause inhibition of tumor cell proliferation, ten N-bromoacetylethanolamine phosphate analogues were designed, synthesized, and tested. They were screened for their activities against various human tumor cell lines to study the effects of inhibition of glycolysis on cell proliferation. The relationship between the structure of these compounds and their inhibitory activity on cell proliferation was also discussed. It was found that the activity of N-(2-methoxyethyl)-bromoacetamide, N-(2-ethoxyethyl)-bromoacetamide, and N-(3-methoxypropyl)-bromoacetamide was comparable to that of the positive control AraC. These three inhibitors showed in vivo anticancer effects in P388 transplant BDF1 mice.

Animals↗

Strongyloidiasis--progress in diagnosis and treatment.

Strongyloidiasis is an intestinal parasitic disease caused by Strongyloides stercoralis. Basically, detecting larvae of S. stercoralis in feces makes definitive diagnosis. The ordinary agar plate culture method developed at our department is much simpler to handle and much more sensitive than the conventional filter paper culture method. It is considered to be the most useful method in the diagnosis of strongyloidiasis and in evaluation of the eradicating effect. Among chemotherapeutic agents, thiabendazole representing the benzimidazole compounds is most effective. However, it has a problem in safety, since its adverse effects and liver dysfunction occur with a high incidence, and it can be severe. Regarding the effects of mebendazole, albendazole and ivermectin, a study was conducted which included many patients. A high incidence of liver dysfunction was observed with mebendazole, and eradicating effect was not sufficient with albendazole. Ivermectin is different from benzimidazole compounds in a pharmacokinetic profile. However, ivermectin showed a strong anthelmintic effect with the least toxicity. We therefore consider ivermectin is the most useful drug for the treatment of strongyloidiasis.

Antiprotozoal Agents↗

Undefined complications of parathyroid adenoma, parathyroid hyperplasia (primary hyperparathyroidism), thyroid follicular adenoma, thyroid papillary carcinoma, temporal astrocytoma, cerebellar meningioma, and hemangioma of external auditory meatus and oral papilloma.

A 59-year-old woman who had parathyroid adenoma, parathyroid hyperplasia, thyroid follicular adenoma, thyroid papillary carcinoma, astrocytoma of the right temporal lobe, cerebellar meningioma, capillary hemangioma of the left external auditory meatus and papilloma of the left upper gingiva is reported. Dynamic magnetic resonance imaging, computed tomography with contrast-enhancement and gastrofiberscopy revealed no remarkable findings in the pituitary, pancreas, adrenals, stomach or duodenum. Similar lesions were not found in any family members. Defect of the causative genes of multiple endocrine neoplasia types I and IIa, MENIN and RET was not detected. Further follow-up of this patient and family members is needed.

Adenoma↗

An anomalous case of the hepato-spleno-mesenteric and the gastro-phrenic trunks independently arising from the abdominal aorta.

This report describes an arterial anomaly case in the celiaco-mesenteric region, which was encountered in a Japanese male cadaver in the dissecting room at Kurume University School of Medicine in 1999. In this case, the usual celiac trunk was not identified, and the hepato-spleno-mesenteric and the gastro-phrenic trunks were independently arising from the abdominal aorta. In addition, the common hepatic artery divided into the left hepatic, the right hepatic, and the gastroduodenal arteries simultaneously, then, the accessory gastric artery arose from the left hepatic artery. This type of arteral anomaly belongs to the Type III of Adachi's classification and the Type II of Morita's classification.

Aged↗

Two anomalous cases of the hepato-mesenteric and the gastro-splenic trunks independently arising from the abdominal aorta.

This report describes two arterial anomaly cases in the celiaco-mesenteric region, which were encountered in two Japanese male cadavers in the dissecting room at Kurume University School of Medicine in 1999. In these cases, the usual celiac trunks were not identified, and the hepato-mesenteric and the gastro-splenic trunks were independently arising from the abdominal aorta. Moreover, in the first case, the common hepatic artery passed ventral side of the portal vein and divided into the hepatic proper and the gastroduodenal arteries. This type of arterial anomaly belongs to the Type V of Adachi's classification and the Type IV' of Morita's classification. In the second case, the common hepatic artery passed dorsal side of the portal vein and divided into the hepatic proper and the gastroduodenal arteries. This type of arterial anomaly belongs to the Type VI of Adachi's classification and the Type IV' of Morita's classification.

Aged↗

Cloning of cDNA encoding ethylene-responsive element binding protein-5 in the cultured cells of Nicotiana tabacum.

We have isolated a full-length cDNA clone from the cultured cells of Nicotiana tabacum, of which the deduced 225 amino acid sequence showed significant homology to the known ethylene-responsive element binding proteins (EREBPs) from tobacco and Arabidopsis. Although the clone shared substantial homology with an Arabidopsis ethylene-responsive element binding factor-3 (AtERF-3) throughout the full length, such a clone had not been found in N. tabacum cells yet; the clone was found to be 5th homologue of EREBPs on N. tabacum.

Amino Acid Sequence↗

[Postoperative interstitial pneumonia: 2 cases after lobectomy].

Two patients with postoperative interstitial pneumonia are reported. Preoperative diagnosis was primary lung cancer without idiopathic interstitial pneumonia (IIP). Within one week after operation acute interstitial pneumonia (AIP) occurred on the nonoperated side and developed. Steroid therapy was performed but one was dead. AIP is a fatal complication after pulmonary resection and steroid therapy may be useful in some cases of postoperative AIP.

Adenocarcinoma↗

[A resected case of large cell neuroendocrine carcinoma].

Large cell neuroendocrine carcinoma (LCNEC) is a rare lung cancer and it has a poor prognosis. We describe our experience with a patient in whom LCNEC was diagnosed. 38-year-old woman who complained of a cough and low grade fever up was admitted to our hospital. Chest X-ray and CT shoued 6.5 x 5.0 mass in rt-S1 and S2. It was suspected as LCNEC of the right lung because of broncopscopic cytology. The upper lobectomy of the right lung and desection of mediastinal lymph nodes were performed. Pathological diagnosis was III B (T2N2M0) LCNEC. Four weeks after the operation, one cycle of chemotherapy (CDDP + VP - 16 + VDS) and one cycle of chemoradiotherapy (thoracic radiation of 40 Gy, CDDP + 5 - FU) were performed. There are no findings of tumor recurrence 7 months after operation.

Adult↗

[A comparative study of the prognosis for Japanese patients with idiopathic interstitial pneumonia or BOOP based on histopathologic subsets].

We explored the prognosis for 123 patients with either idiopathic interstitial pneumonia (IIP) or bronchiolitis obliterans organizing pneumonia (BOOP). All patients underwent either open lung biopsy or thoracoscopic lung biopsy procedures. The histopathologic diagnosis of IIP included patients with usual interstitial pneumonia (UIP), nonspecific interstitial pneumonia (NSIP), and desquamative interstitial pneumonia with respiratory bronchiolitis-associated interstitial lung disease. The prognosis was poorest for patients with a histologic diagnosis of UIP, and excellent for those who received a diagnosis of BOOP. Although the prognosis is generally considered to be good for patients with NSIP, some NSIP patients in our study died. Histopathologic diagnosis based on surgical lung biopsy is useful in evaluating the prognosis for patients with IIP.

Aged↗

[The analysis of surgically treated pulmonary tuberculosis].

Between January 1990 and December 1999, thirteen patients with pulmonary tuberculosis underwent surgical management in our hospital. The purpose of surgery was classified into three groups: drug-resistant or persistent disease (7 patients), hemoptysis (3), and the others. We have no operative death, but have two late deaths due to postoperative persistent positive sputum and progressive tuberculosis infection. One patient relapsed one year and six months after operation and medical treatment was done successfully. Pulmonary resection is useful for localized pulmonary tuberculosis even drug-resistant cases.

Adult↗

Targeted gene disruption demonstrates that P-selectin glycoprotein ligand 1 (PSGL-1) is required for P-selectin-mediated but not E-selectin-mediated neutrophil rolling and migration.

P-selectin glycoprotein ligand 1 (PSGL-1) is a mucin-like selectin counterreceptor that binds to P-selectin, E-selectin, and L-selectin. To determine its physiological role in cell adhesion as a mediator of leukocyte rolling and migration during inflammation, we prepared mice genetically deficient in PSGL-1 by targeted disruption of the PSGL-1 gene. The homozygous PSGL-1-deficient mouse was viable and fertile. The blood neutrophil count was modestly elevated. There was no evidence of spontaneous development of skin ulcerations or infections. Leukocyte infiltration in the chemical peritonitis model was significantly delayed. Leukocyte rolling in vivo, studied by intravital microscopy in postcapillary venules of the cremaster muscle, was markedly decreased 30 min after trauma in the PSGL-1-deficient mouse. In contrast, leukocyte rolling 2 h after tumor necrosis factor alpha stimulation was only modestly reduced, but blocking antibodies to E-selectin infused into the PSGL-1-deficient mouse almost completely eliminated leukocyte rolling. These results indicate that PSGL-1 is required for the early inflammatory responses but not for E-selectin-mediated responses. These kinetics are consistent with a model in which PSGL-1 is the predominant neutrophil P-selectin ligand but is not a required counterreceptor for E-selectin under in vivo physiological conditions.

Animals↗

Expression of proliferating cell nuclear antigen and CD44 variant isoforms in the primary and metastatic sites of nonsmall cell lung carcinoma with intrapulmonary metastases.

BACKGROUND: To the authors' knowledge the prognosis of patients with intrapulmonary metastases (PM) of nonsmall cell lung carcinoma (NSCLC) has not yet been clarified fully and little is known regarding the characteristic changes that occur during the metastatic process, nor of their clinical significance. METHODS: Formalin fixed and paraffin embedded material of primary and metastatic lesions resected from 34 patients with PM of NSCLC were stained immunohistochemically with proliferating cell nuclear antigen (PCNA) and CD44 and its variant isoforms. RESULTS: Patients with NSCLC expressing PCNA in the primary tumor site (79.4%) showed a significantly poorer survival (5-year survival rate of 20.2%) compared with the 5-year survival rate of 57.1% for patients not expressing PCNA in the primary tumor site (P = 0.048). Patients expressing PCNA in the metastatic site (88.2%) also showed a significantly poorer prognosis than those not expressing PCNA (P = 0. 036). Patients with squamous cell carcinoma expressed CD44 variant exon 6 (CD44v6) at a significantly higher rate than adenocarcinoma patients (P = 0.0164), but expression of CD44v6 was not a significant prognostic factor. Concordance of PCNA and CD44v6 expression between the primary and corresponding metastatic sites was observed in 65% of patients (22 of 34 patients) but no difference in prognosis was observed in relation to this concordance. Cox multivariate analyses indicated that expression of PCNA was a significant prognostic indicator for both primary and metastatic sites (P = 0.014 and P = 0.0095, respectively). CONCLUSIONS: This study demonstrated that PCNA expression was a significant prognostic factor for both primary and metastatic lesions in PM patients. CD44v6 showed histogenesis of the tumor, but no relation with the prognosis could be ascertained.

Adenocarcinoma↗

Role of saccharides on lung preservation.

BACKGROUND: Saccharides are considered to play a role as osmotic impermeants and serve as an energy source for the organ during ischemia. However, previous studies on the effectiveness of saccharides on organ preservation have yielded conflicting results. We compared the preservative effects of a monosaccharide (glucose), disaccharides (trehalose, maltose, sucrose), and a trisaccharide (raffinose) to investigate whether the effects of saccharides on lung preservation depend on their molecular weight, energy-level maintenance, and cytoprotective effects. METHODS: We used an ex vivo rat lung model using homologous blood as the perfusate. In the fresh group, the lungs were reperfused immediately after flush. In the other groups, the lungs were flushed with one of the solutions containing glucose, trehalose, maltose, sucrose, or raffinose and preserved for 14 hours. RESULTS: The results of the trehalose group were comparable to those of the fresh group. The glucose, maltose, and raffinose groups showed significantly higher levels of shunt fraction, pulmonary artery pressure, and peak inspiratory pressure compared with the fresh and trehalose groups. There were no differences among the groups in the levels of total adenine nucleotides, adenosine triphosphate of the lung after flush, and preservation. However, after reperfusion, levels of total adenine nucleotides became significantly lower in the glucose, sucrose, maltose, and raffinose groups. Ultrastructural examination revealed endothelial cell injury in the glucose, sucrose, maltose, and raffinose groups. CONCLUSIONS: These results show that the effects of saccharides may depend on their cytoprotective effect rather than on impermeant activity or energy-level maintenance of the preserved lung. Trehalose proved to be superior to the other saccharides.

Adenine Nucleotides↗

Geraniol is a potent inducer of apoptosis-like cell death in the cultured shoot primordia of Matricaria chamomilla.

We screened various isoprenoids to search for inducers of apoptosis-like cell death ("apoptosis") in the shoot primordia of Matricaria chamomilla and found that geraniol has the most potent apoptosis-inducing activity among terpenoids, as judged from DNA fragmentation in the cells. The apoptosis-inducing activity of geraniol is concentration and time dependent. Morphologically, the nuclei of the cells treated with 5 mM geraniol were condensed and fragmented.

Acyclic Monoterpenes↗

Tissue distribution of placenta-type 6-phosphofructo- 2-kinase/fructose-2,6-bisphosphatase.

Several isozymes of 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase have been characterized from mammalian tissues and, based on tissue origin, they are classified as liver, skeletal muscle, heart, testis, and placenta isozymes. In this paper, we examined the tissue distribution of placenta-type isozyme in rat tissues at the levels of transcription and translation. Analysis by Northern blotting showed that placenta, brain, testis, liver, kidney, and skeletal muscle expressed mRNA of placenta-type isozyme. Western blot analysis of fractions from POROS-HQ column chromatography of extracts from various rat tissues showed that proteins of placenta-type isozyme are expressed in placenta, brain, testis, liver, spleen, heart and lung, but not in kidney and skeletal muscle. An immunohistochemical study showed that, in liver, placenta-type isozyme is localized in Kupffer cells. These results indicate that isozymes of this particular enzyme may occur in particular cell types within each tissue.

Animals↗