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Biomedical subjects

T Sakamoto

Publications and source records attributed to T Sakamoto.

At least 505 records · Page 28Linked to original sources

[Surgical treatment of double outlet left ventricle].

Double outlet left ventricle (DOLV) is an uncommon congenital cardiac defect which was first reported by Sakakibara and associates. We studied anatomical aspects relevant to surgical repair in our 4 cases and additional 28 cases which had been reported on the literatures. The locations of ventricular septal defect (VSD) were subaortic in 20 cases, subpulmonary in 7, doubly-committed in 2 and unknown in 2. The positions of the aorta relative to the pulmonary artery were right or left anterior oblique in 11 cases, right or left side-by-side in 9, right or left posterior oblique in 10 and unknown in 3. The definitive surgery included Rastelli type operation in 18 cases, right ventricular outflow tract repair with closure of VSD in 6, intraventricular conduit repair in 4 and Fontan type operation in 4. In regard to the selection of the procedure, the size of right ventricle, the presence of pulmonary stenosis, the location of VSD and the relationship of the great arteries are very important. Although most of the patients underwent the Rastelli type operation, the right ventricular outflow tract reconstruction or intraventricular conduit repair could be adopted particularly in patients with posterior oblique position of the aorta relative to the pulmonary artery or subpulmonary VSD.

Adolescent↗

[Thoracoscopic resection of mediastinal parathyroid adenoma with cyst and hyperparathyroidism--a case report].

A 51-year-old woman was admitted because of general fatigue assumed to be due to climacteric disturbance. Laboratory examinations suggested a diagnosis of hyperparathyroidism based on high serum levels of Ca, intact PTH, and high-sensitivity PTH. No abnormalities were found in the cervix. Chest X-ray films and CT scans showed a mass measuring 4 cm in the right postero-upper mediastinum. The tumor was positive on a parathyroid scintigram, and was diagnosed as a mediastinal parathyroid adenoma with hyperparathyroidism. Operative findings showed a tumor surrounded by the superior vena cava, vagus nerve, azygos vein, and brachiocephalic artery. Careful resection of the hypervascular tumor from surrounding tissues was performed using a thoracoscopic technique alone. The postoperative course was uneventful. Serum values of Ca, intact PTH, and high-sensitivity PTH decreased to within normal ranges. after surgery. Pathohistological study confirmed the diagnosis of parathyroid adenoma including a cyst caused by degeneration of the tumor.

Adenoma↗

Mechanoenergetics of rat left ventricles in in situ and excised blood-perfused hearts and in unloaded rat left ventricular slices.

We investigated rat left ventricular (LV) mechanoenergetics in three different preparations. We obtained an upward convex curvilinear end-systolic pressure-volume relation (ESPVR) regardless of left ventricular (LV) contractility in in situ and excised cross-circulated rat hearts. We also obtained a linear myocardial O2 consumption per beat (VO2)-end-systolic pressure-volume area (PVA; a measure of ventricular total mechanical energy) relation regardless of the curvilinear LV ESPVR in the excised cross-circulated heart. The slope and the VO2 intercept of the VO2-PVA relation was similar to those in other species. The basal metabolism obtained by KCl arrest was higher than those in other species. In the whole heart preparation, the VO2 intercept may include O2 consumption for residual crossbridge cycling as well as O2 consumption for excitation-contraction (E-C) coupling and basal metabolism. Therefore, we established a new system for measuring myocardial O2 consumption of mechanically unloaded (zero PVA) rat LV slices. The increment in O2 consumption by stimulation of the slices represents O2 consumption for E-C coupling (but not for crossbridge cycling). Most O2 consumption for E-C coupling seems consist of O2 consumption for the sarcoplasmic reticulum Ca2+ pump.

Animals↗

[Sero-negative tsutsugamushi disease (scrub typhus) diagnosed by polymerase chain reaction].

We report a case of sero-negative tsutsugamushi disease diagnosed by polymerase chain reaction (PCR). A 54-year-old man who worked in Nagano prefecture presented with flu-like symptoms that did not respond to cephalosporin therapy. On admission to another hospital, chest roentgenography revealed abnormal shadows; liver dysfunction was also present. Despite therapy, the patient's condition gradually worsened and he was transferred to our intensive care unit. Erythema on all extremities and scabs on the right medial femoral region and the dorsum of the left foot suggested a diagnosis of tsutsugamushi disease. We administered minocycline and gave percutaneous cardiopulmonary support for adult respiratory distress syndrome. Despite all efforts, the patient died. Although serologic tests were not positive, Karp strains of R. tsutsugamuschi were identified on PCR amplification. Autopsy revealed evidence of acute hemorrhagic pancreatitis, which has not been reported previously in tsutsugamushi disease. We conclude that PCR techniques may be useful in confirming a diagnosis of early tsutsugamushi disease.

DNA, Bacterial↗

Structure-activity correlation for an HDV ribozyme composed of three RNA strands.

Hepatitis delta virus (HDV) RNA ribozyme system which consists of three RNA oligomer strands (substrate 8-mer; enzyme 16-mer plus 35-mer, Fig. 1) was designed. Effects of Mg2+ concentration on the pseudo first-order rate constant (kobs) of RNA cleavage reaction and on conformation of ribozyme complex were examined. The secondary structure of the complex was also analyzed by limited digestion with ribonucleases. The kobs and CD data were analyzed by curve-fitting analysis using equations derived for two-Mg2+ and three-Mg2+ ion binding models. The result revealed that a three-Mg2+ binding model can explain the Mg(2+)-concentration-dependent changes of both conformation and activity of the HDV ribozyme.

Base Sequence↗

Molecular cloning and nucleotide sequence of the gene encoding phosphate-inducible pectin lyase of Bacillus subtilis.

The gene encoding the pectin lyase (PNL; EC 4.2.2.10) of Bacillus subtilis has been cloned, sequenced, and characterized. A coding sequence for the PNL composed of 345 amino acids including a 24-amino-acid signal peptide was assigned. No sequence resembling a LexA binding site was found upstream of the structural gene. Furthermore, PNL activity of the gene product expressed in Escherichia coli DH5alpha was detected intracellularly, which might suggest that expression of the gene was not controlled by RecA. Regulation of the gene expression seemed to be quite different from that of other bacterial PNL genes previously reported.

Amino Acid Sequence↗

Molecular cloning and cold-inducible gene expression of ferritin H subunit isoforms in rainbow trout cells.

Cold acclimation of rainbow trout cells is considered to be mediated by alterations in the mRNAs and proteins present in cold-treated cells. A subtracted cDNA library from cold-treated rainbow trout RTG-2 cells was constructed and screened to isolate cDNA induced in the cold-treated cells in order to elucidate which genes are induced by cold acclimation. A set of cDNA clones encoding three members of ferritin H isoforms was isolated as cold-inducible genes. Northern blot analysis and nuclear run-on transcription assay showed that the transcription and accumulation of the ferritin H isoforms mRNA were enhanced by cold acclimation. Furthermore, the ferritin level in the trout cells increased on cold acclimation in response to a temperature shift from 22 degrees C to 4 degrees C. When the trout cells were subjected to 4 degrees C under the condition of a decreased ferritin H level obtained by the addition of an antisense oligonucleotide, cell growth was apparently inhibited. These findings indicate an association between the induction of ferritin H and cellular mechanisms during cold acclimation of trout cells.

Amino Acid Sequence↗

Activities of expiratory neurones of the Bötzinger complex during vocalization in decerebrate cats.

Repetitive electrical stimulation of the midbrain peri-aqueductal grey (PAG) terminates quiet breathing and initiates inspiration that precedes vocalization. To understand the neuronal mechanisms underlying this phenomenon, activities of expiratory neurones (n = 39) of the Bötzinger complex (BOT) were examined in decerebrate cats. Most augmenting expiratory (E-aug) neurones (20/22) of the BOT, including 15 bulbospinal neurones, decreased their activities (9/20) or ceased to discharge (11/20) after the onset of stimulation of the PAG. This suggests that suppression of E-aug neurones of the BOT, which project to phrenic motoneurones, results in disinhibition of these neurones, and, in turn, terminates expiration and initiates inspiration preceding vocalization.

Animals↗

Role of tachykinin and bradykinin receptors and mast cells in gaseous formaldehyde-induced airway microvascular leakage in rats.

We have investigated the effects of CP-99,994 [(+)-(2s,3s)-3-(2-methoxybenzylamino)-2-phenylpiperidine], a tachykinin NK1 receptor antagonist, HOE 140 (D-Arg[Hyp3,Thi5,D-Tic7,Oic8]bradykinin), a bradykinin B2 receptor antagonist, and ketotifen (4-(1-methyl-4-piperidylidene)4 H-benzo[4,5]cycloheptal[1,2-b]thiophen-10(9H)-one hydrogen fumarate), a histamine H1 receptor antagonist with mast cell-stabilizing properties, on microvascular leakage induced by gaseous formaldehyde. Extravasation of Evans blue dye into airway tissues was used as an index of airway microvascular leakage. Leakage of dye in the trachea and main bronchi increased significantly in a concentration-dependent fashion after 10 min inhalation of formaldehyde (5-45 parts per million (ppm)). The airway response induced by 10 min inhalation of 15 ppm formaldehyde (trachea: 119.5 +/- 13.9 ng/mg, n = 7; main bronchi: 139.6 +/- 7.9 ng/mg, n = 7) was abolished by the administration of CP-99,994 (3 and 6 mg/kg i.v.), but not by the administration of HOE 140 (0.65 mg/kg i.v.) nor ketotifen (1 mg/kg i.v.). The increase in vascular permeability induced by formaldehyde in the rat airway was mediated predominantly by NK1 receptor stimulation. Activation of bradykinin receptors and mast cells did not appear to play an important role in this airway response.

Animals↗

Recombinant bovine conglutinin, lacking the N-terminal and collagenous domains, has less conglutination activity but is able to inhibit haemagglutination by influenza A virus.

Conglutinin is a bovine serum protein which was first described as a vertebrate lectin. This protein belongs to the family of C-type lectins. These lectins are composed of four characteristic domains: (1) an N-terminal cysteine-rich domain, (2) a collagen-like domain, (3) a neck domain and (4) a carbohydrate recognition domain (CRD). Recently lectins have been shown to function as immunoglobulin-independent defence molecules due to a complement-mediated mechanism or opsonization. Our previous study showed that bovine conglutinin can inhibit haemagglutination by influenza A viruses and act by directly neutralizing them due to its lectin properties. In order to elucidate the biological role of the collagen-like domain, a recombinant partial conglutinin lacking this collagen-like domain was produced in an Escherichia coli system and its biological activities were examined. A 497 bp sequence, consisting of a short collagen region (two repeats of G-X-Y amino acid sequences), the neck domain, and the CRD of conglutinin cDNA, was amplified by the reverse-transcriptase PCR technique. The cDNA was transferred to a bacterial expression vector system (pRSET-A) and stable transfectants with a high level of conglutinin production were obtained. SDS/PAGE and Western blotting analyses showed a recombinant fusion protein of 27 kDa. Results of a cross-linking study and gel-filtration assay indicated that the recombinant conglutinin can form a trimeric structure and that it has sugar binding activity and specificity similar to that of native conglutinin. The recombinant conglutinin was also found to inhibit haemagglutination caused by influenza A virus as well as to possess less conglutination activity. These results suggest that in order for conglutinin to inhibit haemagglutination caused by the influenza virus, as well as to have sugar binding activity or to form trimers, it does not require the N-terminal and collagenous domains; however, they are essential for full conglutination activity.

Amino Acid Sequence↗

Retrovirus-mediated suicide gene transduction in the vitreous cavity of the eye: feasibility in prevention of proliferative vitreoretinopathy.

In proliferative vitreoretinopathy (PVR), retinal pigment epithelial cells, fibroblasts, or other proliferating cells form contractile membranes in the vitreous cavity of the eye, resulting in traction retinal detachment. Retroviral vector-mediated transfer is a suitable method to transduce the herpes simplex virus thymidine kinase (HSV-tk) gene into proliferating cells in PVR, allowing for the selective killing of these cells. To determine the potential of gene transduction in the environment of the vitreous cavity, we evaluated the effect of vitreous humor on retroviral vector-mediated gene transduction of rabbit dermal fibroblasts in vitro and studied in vivo transduction in rabbit experimental PVR with retroviral vector G1BgSvNa. In addition, we studied the bystander effect in vitro and in vivo in a rabbit model of PVR, with low percentages of HSV-tk-positive cells. Finally, we evaluated the efficacy of intravitreal administration of HSV-tk retroviral vector G1TkSvNa followed by ganciclovir (GCV) in the prevention of experimental PVR. Vitreous humor reduced gene transfer efficiency in vitro in a dose-dependent manner. LacZ expression was found in cells of preretinal or intravitreal membranes of animals of both in vivo and in vitro transduction groups; however, in vivo transduction resulted in a decreased number of transduced cells, with a relative transduction efficiency of approximately 2%. Transduction of HSV-tk was associated with a powerful bystander effect both in vitro and in vivo with significant effects even when HSV-tk-positive cells represented only 1% of the population. In vivo transduction with G1TkSvNa followed by GCV significantly inhibited the development of PVR (p < 0.05). These results suggest that retroviral vector-mediated transfer of HSV-tk into the proliferating cells in PVR may be feasible and may provide a novel therapeutic strategy for this disease.

Animals↗

Modulation of cell death pathways to apoptosis and necrosis of H2O2-treated rat thymocytes by lipocortin I.

Lipocortin I, also called annexin I, a calcium and phospholipid binding protein, protected rat thymocytes from H2O2-elicited necrosis and facilitated H2O2-induced apoptosis, while anti-lipocortin I antibody enhanced H202-elicited necrosis by blocking H202-induced apoptosis. Essentially similar results were obtained with phospholipase A2 inhibitors and activators such as 3,4-octyadecyl-benzylacrylic acid and melittin, respectively. Available evidence suggests that lipocortin I modulates signals for cell death pathways of H2O2-treated rat thymocytes to apoptosis and necrosis by regulating cellular phospholipase A2 activities but not by inhibiting membrane lipid peroxidation.

Animals↗

Induction of proteins in response to cold acclimation of rainbow trout cells.

The in vitro translation of poly(A)-RNA isolated from control and cold-treated cells showed that low temperatures induced changes in the population of translatable mRNAs. When cellular proteins extracted from cold-treated cells were subjected to two-dimensional gel electrophoresis, the 70 kDa protein was found to be synthesized during the cold treatment. N-terminal sequence analysis showed that the 70 kDa cold-inducible protein was a homolog of the mammalian valosin-containing protein and yeast CDC48p. The changes in mRNA and protein contents during cold acclimation may result from the expression of genes involved in the adjustment of cellular metabolism to low temperature or the induced proteins may be directly involved in the cold acclimation.

Acclimatization↗

Role of bradykinin B2 receptors and mast cells in the bradykinin-induced skin response in the rat.

We investigated the role of activation of bradykinin receptors and mast cells in the microvascular leakage of the vessels of the skin induced by the intracutaneous (i.c.) injection of bradykinin in the rat. We evaluated the effects of HOE140 (D-Arg-[Hyp3,Thi5,D-Tic7,Oic8]bradykinin), a bradykinin B2 receptor antagonist, and ketotifen (4-(1-methyl-4-piperidylidene)4H-benzo[4,5]cyclohepta[1, 2-b]thiophen-10(9H)-one hydrogen fumarate), a histamine H1 receptor antagonist with mast cell stabilizing properties, on the skin response. Evans blue dye extravasation served as an index of the increase in vascular permeability. Bradykinin (2-100 nmol/site i.c.) induced the extravasation of Evans blue dye in a dose-dependent manner. Ketotifen (20 mg/kg i.p.) significantly inhibited the leakage of dye induced by bradykinin (10 nmol/site i.c.) by 66.2%, while HOE140 (1 mg/kg i.v.) had no effect. The concomitant injection of HOE140 (0.2, 2 nmol/site) and bradykinin (10 nmol/site i.c.), also did not significantly reduce the extravasation of dye. We conclude that the extravasation of plasma induced by the i.c. injection of bradykinin is mediated mainly by stimulation of the skin mast cells, but not by bradykinin B2 receptors.

Animals↗

Poor correlation with loss of heterozygosity on chromosome 17p and p53 mutations in ovarian cancers.

To define the target of chromosome 17p deletions, allelic losses in the 17p11.2 to 13.3 regions of 32 ovarian cancers were investigated. Twenty-one (68%) of 31 informative cancers had deletions on chromosome 17p. None of these 21 cancers involved deletions in the entire chromosome 17p even if deletions of a small chromosome region were infrequent. Of these 21, 17 cancers contained deletions at 17p13.1 or neighboring regions. The remaining 4 cancers with 17p deletions were uninformative for deletions at 17p13.1. Thus, most 17p deletions seemed to target the 17p13.1 region in ovarian cancers. Of the 30 ovarian cancers screened, 6 contained p53 mutations. One p53 allele was lost as a consequence of deletion and the other was mutated in 4 cancers. Seventeen cancers with deletions on 17p showed no evidence of p53 mutations. Thus, deletions on 17p that are common in ovarian cancers are not always accompanied by p53 gene mutations.

Alleles↗