Biomedical subjects

H Sakamoto

Publications and source records attributed to H Sakamoto.

At least 163 records · Page 9Linked to original sources

PubMed1999

Induction of apoptosis and suppression of clonogenicity of ovarian carcinoma cells with estrogen mustard.

BACKGROUND: This study was conducted to evaluate whether estramustine (estrogen mustard [EM]) is a promising alternative in the treatment of patients with epithelial ovarian carcinoma (OVCA). EM is a microtubule-associated protein (MAP) specific antimicrotubule agent with low toxicity. METHODS: The authors investigated the ability of EM to induce apoptosis and suppress colony formation of OVCA cells. Paclitaxel was used as a positive control. DNA electrophoresis and terminal deoxynucleotidyl dUTP-X nick end labeling (TUNEL) assays were used to detect internucleosomal DNA fragmentation. Flow cytometry was used to identify apoptotic cells and disturbance of the cell cycle of EM-treated OVCA cells further. Quantitation of detached cultured cells also provided a relative estimate of the apoptotic response of OVCA cells to treatment with EM. The colony formation assay was used to evaluate the effects of EM on clonogenicity. RESULTS: The effects of EM on four OVCA cell lines in culture were highly similar to those of paclitaxel in causing apoptosis and inhibiting clonogenicity. DNA electrophoresis and TUNEL assays showed that EM induced internucleosomal DNA fragmentation in OVCA cells. Flow cytometry showed changes typical of apoptotic changes and cell cycle block and synchronization at the G2/M-phase. Counting of detached cells showed a log-dose response to EM treatment. The colony formation assay also showed a log-dose response suppression of OVCA cell clonogenicity after treatment with EM. CONCLUSIONS: EM may be a promising candidate in the clinical treatment of patients with OVCA. The lower toxicity and MAP specific action of EM is a novel chemotherapy for OVCA.

Antineoplastic Agents, Alkylating↗

PubMed1999

Naturally occurring somatic motoneuron death in a teleost angelfish, Pterophyllum scalare.

Naturally occurring somatic motoneuron death in a teleost angelfish, Pterophyllum scalare, was investigated histochemically and electron microscopically. The number of motor axons in the ventral root, which corresponds to the motoneuron number in spinal hemisegment, was rapidly increased beyond the adult value within 3 days after hatching, and then decreased to reach the adult value within a few weeks. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) histochemistry, which detects fragmented nuclear DNA characteristic to apoptotic cells, showed that the apoptotic cells are located in the motor column of the cord in the larvae at specific developmental stages. Electron microscopic observations of the spinal cells further confirmed the motoneuron apoptosis. The present data suggest that the massive death of somatic motoneurons at certain ontogenic stages which has been known to occur in higher vertebrates also takes place in fish.

Animals↗

PubMed1999

Identification of a novel promoter and exons of the c-ERBB-2 gene.

The c-ERBB-2 gene is frequently amplified in various cancers. During the course of studies on the structural characterization of the amplification units, we isolated four cDNA clones, A39, GRB7, C51 and CAB1 with corresponding genes localized on the c-ERBB-2 locus. A tentative gene, C51, was located at about 12 kb upstream of the c-ERBB-2 gene. By molecular cloning of a full-length cDNA clone of C51 and the reverse transcription PCR (RT-PCR) method, we identified a novel transcript of c-ERBB-2 containing new 5' sequences including the C51 sequence, demonstrating that the c-ERBB-2 gene has a novel promoter and new exons. The structural organization of the novel promoter and exons of c-ERBB-2 was revealed by complete sequence analysis of a total size of about 20 kb of genomic DNA clones containing the 5'-flanking region of the previously described c-ERBB-2 gene. Transient expression of the newly identified promoter-reporter gene constructs in breast cancer cell line MCF-7 showed that the elements responsible for promoter activity were contained in a 697 bp region upstream of the transcriptional start site. The new transcript may encode a protein different in the portion of the extracellular domain. Although the presence of the predicted protein product was not examined, this report is important in that it provides a new aspect of the c-ERBB-2 protooncogene products.

Adult↗

PubMed1999

Structural basis for recognition of the tra mRNA precursor by the Sex-lethal protein.

The Sex-lethal (Sxl) protein of Drosophila melanogaster regulates alternative splicing of the transformer (tra) messenger RNA precursor by binding to the tra polypyrimidine tract during the sex-determination process. The crystal structure has now been determined at 2.6 A resolution of the complex formed between two tandemly arranged RNA-binding domains of the Sxl protein and a 12-nucleotide, single-stranded RNA derived from the tra polypyrimidine tract. The two RNA-binding domains have their beta-sheet platforms facing each other to form a V-shaped cleft. The RNA is characteristically extended and bound in this cleft, where the UGUUUUUUU sequence is specifically recognized by the protein. This structure offers the first insight, to our knowledge, into how a protein binds specifically to a cognate RNA without any intramolecular base-pairing.

Amino Acid Sequence↗

PubMed1999

The JAK-binding protein JAB inhibits Janus tyrosine kinase activity through binding in the activation loop.

The Janus family of protein tyrosine kinases (JAKs) regulate cellular processes involved in cell growth, differentiation and transformation through their association with cytokine receptors. However, compared with other kinases, little is known about cellular regulators of the JAKs. We have recently identified a JAK-binding protein (JAB) that inhibits JAK signaling in cells. In the studies presented here we demonstrate that JAB specifically binds to the tyrosine residue (Y1007) in the activation loop of JAK2, whose phosphorylation is required for activation of kinase activity. Binding to the phosphorylated activation loop requires the JAB SH2 domain and an additional N-terminal 12 amino acids (extended SH2 subdomain) containing two residues (Ile68 and Leu75) that are conserved in JAB-related proteins. An additional N-terminal 12-amino-acid region (kinase inhibitory region) of JAB also contributes to high-affinity binding to the JAK2 tyrosine kinase domain and is required for inhibition of JAK2 signaling and kinase activity. Our studies define a novel type of regulation of tyrosine kinases and might provide a basis for the design of specific tyrosine kinase inhibitors.

Amino Acid Sequence↗

PubMed1999

APS, an adaptor protein containing PH and SH2 domains, is associated with the PDGF receptor and c-Cbl and inhibits PDGF-induced mitogenesis.

Previously we cloned a novel adaptor protein, APS (adaptor molecules containing PH and SH2 domains) which was tyrosine phosphorylated in response to c-kit or B cell receptor stimulation. Here we report that APS was expressed in some human osteosarcoma cell lines, markedly so in SaOS-2 cells, and was tyrosine-phosphorylated in response to several growth factors, including platelet derived growth factor (PDGF), insulin-like growth factor (IGF), and granulocyte-macrophage colony stimulating factor (GM-CSF). Ectopic expression of the wild type APS, but not C-terminal truncated APS, in NIH3T3 fibroblasts suppressed PDGF-induced MAP kinase (Erk2) activation, c-fos and c-myc induction as well as cell proliferation. In vitro binding experiments suggest that APS bound to the beta type PDGF receptor, mainly via phosphotyrosine 1021 (pY1021). Indeed, tyrosine phosphorylation of PLC-gamma, which has been demonstrated to bind to pY1021, but not that of PI3 kinase and associated proteins, was reduced in APS transformants. PDGF induced phosphorylation of the tyrosine residue of APS close to the C-terminal end. In vitro and in vivo binding experiments indicate that the tyrosine phosphorylated C-terminal region of APS bound to c-Cbl, which has been shown to be a negative regulator of tyrosine kinases. Since coexpression of c-Cbl with wild type APS, but not C-terminal truncated APS, synergistically inhibited PDGF-induced c-fos promoter activation, c-Cbl could be a mechanism of inhibitory action of APS on PDGF receptor signaling.

Adaptor Proteins, Signal Transducing↗

PubMed1999

Enhancement of glucuronosyl etoposide transport by glutathione in multidrug resistance-associated protein-overexpressing cells.

Multidrug resistance-associated protein (MRP) has been shown to transport glutathione (GSH) S-conjugates such as leukotriene C4 (LTC4) and S-(2,4-dinitrophenyl)-glutathione (DNP-SG). On the other hand, it has while it has been reported that MRP-overexpressing cells exhibit decreased sensitivity to drugs which do not form GSH S-conjugates. In this study, we found that GSH affects the transport of glucuronosyl etoposide as a major metabolite of etoposide in MRP-overexpressing KB/VP-4 cells. The relative resistance level of KB/VP-4 cells to etoposide was 70-fold that of wild-type KB cells. Membrane vesicles prepared from KB/VP-4 cells exhibited markedly enhanced ATP-dependent transport of glucuronosyl etoposide as well as LTC4. Transport of glucuronosyl etoposide was augmented in the presence of GSH. Treatment of KB/VP-4 cells with buthionine sulfoximine (BSO), an inhibitor of GSH synthesis, resulted in about 75% depletion of cellular GSH levels, a four-fold increase of the sensitivity to etoposide and depression of glucuronosyl etoposide efflux. These results suggest that GSH plays a role in the enhancement of MRP-mediated glucuronosyl etoposide transport.

ATP-Binding Cassette Transporters↗

PubMed1999

Accessory cells with immunophenotypic and functional features of monocyte-derived dendritic cells are recruited to the lung during pulmonary inflammation.

Pulmonary macrophages (Mphi) increase in tissue and bronchoalveolar lavage (BAL) fluid during inflammation caused by bleomycin (BLM). This study demonstrates that increasing numbers of exudate Mphi in BLM lung injury exhibit dendritic cell (DC) features. After the intratracheal administration of BLM (0.075 U), adherent mononuclear cells from the bronchoalveolar lavage fluid (BAMC) of C57BL/6 mice were characterized for morphology, immunophenotype, and accessory cell activities. At day 7 post-BLM, 48% of CD11b+ BAMC displayed features of DC differentiation, as judged by dendritic morphology, expression of class II MHC, 33D1, Factor XIIIa, CD80, and CD86 antigens, and the ability to support a primary allogeneic lymphocyte response (MLR). After BLM treatment, CD11b+ peripheral blood monocytes also showed increased expression of 33D1, Factor XIIIa, CD86, and the ability to stimulate an MLR. We conclude that inflammatory DC with immunophenotypic features of monocyte-derived DC increase in the peripheral blood and lung after an inflammatory stimulus.

Animals↗

PubMed1999

Fish calcitonin genes: primitive bony fish genes have been conserved in some lower vertebrates.

Using the polymerase chain reaction method, we amplified calcitonin genes from the cDNA of the ultimobranchial glands or from the genomic DNA of the blood cells and liver of various fishes. The fishes examined were primitive bony fishes (lungfish, polypterus, sturgeon, and gar), 16 species of teleosts, and cartilaginous fishes (stingray). Sequenced calcitonin genes were compared among fishes and with those of reptiles and chickens. The similarity of the calcitonin genes was the highest between the reptile and chicken groups and the primitive bony fishes (sequence similarity of the nucleotides 81-90%). The values between teleosts and the primitive bony fishes were 70-81%, with the exception of eels. Eel calcitonin genes were very similar to those of primitive bony fishes (83-88%). Stingray calcitonin genes were relatively more similar to those of the primitive bony fishes (74-78%) than to teleosts (63-73%). In goldfish and sardine, two types of calcitonin genes were found. We concluded that the genes of primitive bony fish are placed at a fundamental position in this hormone, at least among these vertebrates.

Animals↗

PubMed1999

Thymidine phosphorylase expression is predominantly observed in stroma of well-differentiated adenocarcinoma of endometrium and correlates with a frequency of vascular involvement.

Thymidine phosphorylase (dThdPase) expression was studied in 20 cases of well-differentiated endometrial carcinomas to examine the clinicopathological significance. Immunohistochemical study showed predominant dThdPase expression in tumor stroma. No expression was detected in coexisting normal endometrial stroma or stroma of endometriosis. Seven cases (35%) showed strong stain and 13 (65%) showed weak stain. Based on the strength of immunoreactivity, patients were grouped into two groups: group S (strong) and group W (weak). In group S, 5 (71%) showed vascular involvement whereas in group W, only 3 (23%) cases were positive, giving a significant difference in the frequency of vascular involvement between the two groups (P < 0.03, chi2 analysis). No correlation was found between dThdPase expression and myometrial invasion. Patients with vascular involvement resulted in poorer outcome (P < 0.003) whereas between group S and W, there showed no difference in survival (group S vs group W; P < 0.15). A multivariate analysis including stage, vascular involvement, and dThdPase stain as variables showed none as independent risk factors. However, univariate analysis showed that the presence of a vascular involvement was a risk factor for a shorter survival (relative risk = 6.277, 95% range = 1.2-32.6, P < 0. 03). It is concluded that expression of dThdPase in endometrial carcinoma is a marker of a desmoreaction and a risk factor for vascular invasion. Since vascular invasion is a risk factor for poor outcome, the significance of dThdPase expression in endometrial carcinoma requires further clinical evaluations.

Adenocarcinoma↗

PubMed1999

An anomalous right gastroepiploic artery arising from the superior mesenteric artery.

During a routine dissection, the right gastroepiploic artery was found to arise from the superior mesenteric artery. The gastroduodenal artery ran in front of the common bile duct and descended along the posterior surface of the head of the pancreas (posterior superior pancreaticoduodenal artery). The enlarged pancreatic branch arising from the superior mesenteric artery mainly supplied the anterior surface of the head of the pancreas and then continued to become the right gastroepiploic artery. This route seemed to be formed due to the lack of connection between the posterior superior pancreaticoduodenal artery and the common trunk of the anterior superior pancreaticoduodenal and right gastroepiploic arteries.

Aged↗

PubMed1999

Intraoperative irradiation after surgery for locally recurrent rectal cancer.

PURPOSE: This study retrospectively evaluated the effects of intraoperative electron beam irradiation on patients with locally recurrent (pelvic) rectal cancer. METHODS: From November 1, 1975, to December 31, 1997, 51 patients underwent surgery for locally recurrent rectal or rectosigmoid cancer, and 27 patients received intraoperative electron beam irradiation. The intraoperative electron beam irradiation dose was 15 to 30 Gy. Kaplan-Meier survival estimates at three and five years were analyzed for the 47 patients who recovered postoperatively. RESULTS: Statistically significant factors related to survival included intraoperative electron beam irradiation vs. no intraoperative electron beam irradiation (P=0.0007), amount of residual tumor (slight vs. gross; P=0.0022), and symptom status (P=0.0024). Factors not associated with survival included distant metastases at reoperation, type of surgery for the recurrent tumor, external beam irradiation, pathologic grade, age, and gender. Surgical resection without intraoperative electron beam irradiation resulted in three-year and five-year survival rates of 5 and 0 percent, respectively. For patients who received intraoperative electron beam irradiation, the three-year survival rate was 43 percent and five-year survival rate was 21 percent. Intraoperative electron beam irradiation was a statistically significant factor related to survival in patients with and without distant metastasis (P=0.04 and P=0.0035, respectively), with slight residual tumor (P=0.0003), or with palliative surgery (P=0.0276). CONCLUSION: The trends seen in resection with intraoperative electron beam irradiation are encouraging with regard to improvements in survival as compared with studies not using intraoperative electron beam irradiation treatment.

Adult↗

PubMed1999

Two cases of chronic tonsillitis studied by FDG-PET.

We report two cases of chronic tonsillitis studied by FDG-PET. Symmetrical high FDG uptake by the tonsils was observed in both cases. On histopathologic examination of the resected tonsils, follicular hyperplasia was observed with proliferation of lymphocytes in the germinal centers. Increased glucose metabolism in active inflammation involving lymphocyte proliferation was thought to be a cause of high FDG uptake by tonsils in chronic tonsillitis.

Adolescent↗

PubMed1999

The passage of blood-borne horseradish peroxidase into the amygdaloid area of the mouse brain.

The passage of blood-borne horseradish peroxidase (HRP) into the amygdaloid area of the mouse brain was examined with light and electron microscopy. Staining reaction for HRP appeared in medial portions of the amygdaloid area, especially adjacent to the optic tract. Ultrastructural examination of some vessels in that area revealed that the staining reaction for HRP appeared in the perivascular space, the basal lamina, the cytoplasm or vesicular structures of the perivascular cells, vesicular profiles of the endothelial cell cytoplasm including abluminal pits and the adjacent extracellular space. These findings suggest that intravascular macromolecules can invade medial portions of the amygdaloid area of the mouse brain.

Amygdala↗

PubMed1999

Motor innervation by enteric nerve fibers containing both nitric oxide synthase and galanin immunoreactivities in the striated muscle of the rat esophagus.

The relationship between nitric oxide synthase (NOS)- and galanin-immunoreactive nerve terminals and the origin of NOS-immunoreactive nerve terminals on the motor endplates in the striated muscles of the rat esophagus was investigated. Double immunohistochemical staining revealed a dual innervation of motor endplates by calcitonin gene-related peptide (CGRP)-immunoreactive axons and by axons that were immunoreactive for both NOS and galanin. On average, 91% of NOS terminals were galanin immunoreactive. NOS-immunoreactive fibers were revealed at 67% of endplates, identified by the presence of CGRP terminals. The left vagus and superior laryngeal nerve were cut and 15 days allowed for terminals to degenerate. This caused a significant loss of CGRP fibers, but did not affect the density of innervation of the striated muscle by NOS-immunoreactive fibers. Thus the NOS/galanin fibers are deduced to originate from ganglia in the esophageal wall. This is supported by our observation of numerous NOS-immunoreactive nerve cell bodies in the myenteric ganglia of the esophagus, 74% of which were galanin immunoreactive. There were no CGRP-immunoreactive nerve cell bodies in the wall of the esophagus.

Animals↗

PubMed1999

Expansive suboccipital cranioplasty for the treatment of syringomyelia associated with Chiari malformation.

In order to treat syringomyelia associated with adult type Chiari malformation, the authors developed a method of expansive suboccipital cranioplasty (ESC) that involves enlarging the small posterior fossa to obtain a sufficient flow of cerebrospinal fluid (CSF). The relative effectiveness of ESC with the obex plugged and not plugged was also examined, as well as other factors influencing the operative results. Twenty patients without arachnoid adhesion at the major cistern underwent ESC without opening the arachnoid membrane at the major cistern. After surgery, all improved with no recurrence and CSF flow study using magnetic resonance (MR) imaging showed significant improvement of the flow at the major cistern. Another 20 patients without arachnoid adhesion also underwent ESC but with obex plugging. Sixteen improved and one displayed only temporary improvement with recurrent syringomyelia due to postoperative arachnoid adhesions. The remaining three showed no change in spite of shrinkage of the syrinx on postoperative MR imaging. These three patients had displayed pre-operative symptoms over an approximately 10-year period involving almost the entire axial plain of the spinal cord, and presented a large syrinx before surgery. In 4 patients with arachnoid adhesions, all required intra-arachnoid procedures in addition to ESC. Intra-arachnoid procedures are not necessary to facilitate restoration of CSF flow in patients without arachnoid adhesions, because ESC can release the CSF flow blockage in the major cistern even without plugging of the obex. An associated arachnoid adhesion at the major cistern or a long-standing syringomyelia with irreversible damage of the spinal cord results in a poor operative prognosis. When posterior fossa surgery fails, insufficient decompression or postoperative arachnoid adhesions at the major cistern as the cause of treatment's failure should be evaluated by CSF flow studies using phase contrast MR imaging.

Adult↗

PubMed1999

Isolation of bacteria from cervical lymph nodes in patients with oral cancer.

Thirty patients with oral mucosal cancer were studied in relation to oral mucosal damage and bacterial translocation to the regional lymph nodes in the neck. All 30 patients (gingiva 11, tongue 13, cheek mucosa four, oral floor two) underwent extensive, clean-contaminated, head-and-neck surgery (including neck dissection) with free flap reconstruction. A total of 153 lymph nodes was harvested for microbial and histological examination. Viable bacteria were isolated from 70 lymph nodes (45.8%) from 25 patients (83.3%). Bacterial cells in the nodes were detected by scanning electron microscopy. Bacterial translocation was found more often in metastatic nodes (75.0%) than in uninvolved nodes (40.3%) (p = 0.015; chi2 test). Gingival carcinoma yielded 56.4% of bacterial growth in the regional lymph nodes compared to tongue (40.3%), oral floor (40.0%) and cheek mucosa (37.5%). As the gingival carcinoma group includes more T4 cases (11/11; 100%) than the other three groups (7/19; 36.8%), bacterial translocation in uninvolved nodes could be caused by the size and invasion of the primary oral tumor. Oral streptococci (Streptococcus intermedius, Strep. constellatus, Strep. oralis, Strep. mitis, Strep. sanguis, Strep. salivarius) were the most common isolates. Aerobic enteric bacteria (Enterococcus, Escherichia, Klebsiella etc.) were also found in the lymph nodes. Among the anaerobic bacteria, Peptostreptococcus spp. were isolated from 12 patients. Damaged oral mucosa in patients with oral cancer might allow the new bacterial colonization on the surface and subsequently drain the bacteria into the regional lymph nodes as well as the general circulation.

Bacteria↗

PubMed1999

"White coat effect" induced by therapist's presence during speech therapy for stroke rehabilitation: a single case study.

The excessive pressor response triggered in patients by an alerting reaction to a doctor's presence has been termed the "white coat effect." A 68-year-old man with verbal apraxia after multiple lacunar infarctions was referred to the hospital for speech rehabilitation. He experienced difficulty in talking with the speech therapist during therapy sessions but not when talking with his friends or family. Because the therapist's presence was stressful to the patient, it was considered that his anxiety might produce an excessive increase in blood pressure. Blood pressure monitoring was performed during 2 separate days of speech therapies consisting of two sessions each. In one session, therapy was directed by the therapist; in the other, therapy was self-directed. The therapist-directed approach substantially increased both systolic and diastolic blood pressures, whereas the self-directed therapy slightly increased only systolic pressure. It was concluded that the excessive pressor response seen in this patient during therapist-directed speech therapy resulted from the white coat effect induced by the therapist's presence.

Aged↗