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Biomedical subjects

T Sugimoto

Publications and source records attributed to T Sugimoto.

At least 379 records · Page 21Linked to original sources

Synthetic of Uniform Spindle-Type Titania Particles by the Gel-Sol Method

Uniform spindle-type titania (TiO2) particles were prepared by a novel gel-sol method. A mixed solution of titanium(IV) isopropoxide and triethanolamine at molar ratio 1:2 prepared in a drybox was mixed with doubly distilled water to make the final concentration of titanium(IV) 0.50 mol dm-3. The mixture was further mixed with an equal volume of 2.0 mol dm-3 ammonia solution, and the resulting solution was aged for 24 h at 100°C. By this aging a highly viscous gel was formed, but there was no sign of nucleation of titania at this stage. The gel was then aged at 140°C for 3 days to nucleate and grow the titania particles. The gel network formed in the first aging played a decisive role in the second aging to produce the uniform particles of titania, as a reservoir of the metal ions to lower the supersaturation for preventing extensive nucleation and as an anticoagulant fixing the growing particles in the matrix. The product was identified by XRD as anatase particles elongated in the direction of the c-axis. The aspect ratio, as well as the size of the resulting particles, was reduced as the concentration of ammonia was decreased. Copyright 1997Academic Press

Journal Article↗

Stimulatory effects of parathyroid hormone and 1,25-dihydroxyvitamin D3 on insulin-like growth factor-binding protein-5 mRNA expression in osteoblastic UMR-106 cells: the difference between transient and continuous treatments.

The transient treatment with parathyroid hormone (PTH) for 12 h, followed by its removal for 36 h, stimulated insulin-like growth factor-binding protein (IGFBP)-5 mRNA expression more strongly than the continuous treatment for 48 h in osteoblastic UMR-106 cells. The transient but not continuous treatment with A23187 also stimulated it. In contrast, 1,25-dihydroxyvitamin D3 stimulated it, irrespective of the treatment design. IGFBP-5 stimulated type-1 procollagen mRNA expression. The present study first indicated that the transient treatment with PTH more effectively stimulated IGFBP-5 mRNA expression than its continuous treatment partly via an increase in intracellular calcium and suggested that IGFBP-5 might be involved in the anabolic action of PTH in bone.

Animals↗

1-desamino-8-D-arginine vasopressin (DDAVP) as an agonist on V1b vasopressin receptor.

1-desamino-8-D-arginine vasopressin (DDAVP) is considered a standard vasopressin V2 receptor-selective agonist with a potent antidiuretic effect through V2 receptor without the induction of vasoconstriction through V1a receptor. Furthermore, DDAVP was reported to act as an agonist on non-V1a, non-V2 receptor to cause the accumulation of intracellular Ca2+ in several tissues. However, the agonistic activity of DDAVP against the other vasopressin receptor, V1b (or V3), which can accumulate intracellular Ca2+ and which we recently cloned, has not been clarified. Hence, we compared the characteristics of DDAVP on V1b receptor with those on the other vasopressin receptors. In binding experiments, DDAVP more strongly inhibited [3H]arginine vasopressin binding to V1b than to V2 receptor (Ki: 5.84 nM vs 65.9 nM). In addition, DDAVP dose-dependently stimulated inositol turnover in human V1b receptor-expressing COS-1 cells. DDAVP acted as a full agonist on human V1b receptor (EC50: 11.4 nM) as well as on human V2 receptor (EC50: 23.9 nM). However, DDAVP behaved as a partial agonist toward rat V1b receptor (intrinsic activity: 0.7, EC50: 43.5 nM), while there was no significant difference in the agonistic properties of arginine vasopressin on human and rat V1b receptor. In conclusion, DDAVP acts as an agonist on V1b receptor, as it does on V2 receptor. These findings will allow us to better understand the physiological role of V1b receptor in pancreatic beta cells and in the renal inner medullary collecting duct, and help us to identify as yet unknown vasopressin receptors through which DDAVP cause the accumulation of intracellular Ca2+ in other tissues.

Animals↗

Cloning of a mRNA preferentially expressed in chondrocytes by differential display-PCR from a human chondrocytic cell line that is identical with connective tissue growth factor (CTGF) mRNA.

Chondrocyte- or chondrosarcoma cell line (HCS)-specific DNA fragments were obtained using differential display-PCR. Nucleotide sequences of 32 species derived from HCS cells were determined. One of the sequence tags (tag no. 24) corresponded to the nucleotide sequence of connective tissue growth factor (CTGF). Northern blot analysis showed that CTGF was highly expressed in HCS cells and rabbit growth cartilage cells in culture but was not expressed in osteoblastic cells in culture. In situ hybridization revealed that CTGF was expressed only in the hypertrophic chondrocytes of costal cartilage and the vertebral column in embryonic mice. The expression of CTGF in HCS cells was up-regulated by the addition of TGF-beta or BMP-2. These findings suggest that CTGF participates in endochondral ossification.

Animals↗

Unrestrained nociceptive response and disregulation of hearing ability in mice lacking the nociceptin/orphaninFQ receptor.

In the G-protein-coupled receptor superfamily, the opioid receptor subfamily is constituted of the three distinct opioid receptors (namely delta-, mu- and kappa-subtypes) and the receptor for nociceptin (also designated orphaninFQ). The members of the opioid receptor subfamily were known to mediate a variety of cellular inhibitory effects. The three opioid receptors are known to play central roles in mediating analgesia and many other physiological activities; however, the nociceptin receptor was identified recently and less is known about its physiological roles. Here we report the generation and characterization of mice lacking the nociceptin receptor. The knockout mice showed no significant differences in nociceptive threshold and locomotor activity compared with control mice, but they lost nociceptin-induced behavioral responses. These results indicate that the nociceptin system is not essential for regulation of nociception or locomotor activity. On the other hand, we found insufficient recovery of hearing ability from the adaptation to sound exposure in the mutant mice. Thus, the nociceptin system appears to participate in the regulation of the auditory system.

Animals↗

Interference of a methotrexate derivative with urinary oncopterin [N2-(3-aminopropyl)biopterin] measurement by high-performance liquid chromatography with fluorimetric detection.

We previously reported a HPLC assay method using fluorimetric detection for the simultaneous determination of urinary N2-(3-aminopropyl)biopterin (oncopterin, a natural pteridine newly found in urine from cancer patients), biopterin and neopterin. We now have observed that an unknown substance, which may be derived from methotrexate, in urine from a patient with stomach cancer interfered with the assay of oncopterin and demonstrated that oncopterin could be completely separated from the unidentified substance by HPLC using a Nucleosil 100-5SA strong cation-exchange column. Furthermore, oncopterin was not detectable by this HPLC-fluorimetric method in urine samples from patients with stomach cancer who were not treated with methotrexate. The content of urinary oncopterin from cancer patients is supposed to be very low, with less than 1 mumol/mol creatinine. The present results indicate that the peak found with elution from the C18 column was a methotrexate-derived compound and co-eluted with the analyte oncopterin.

Biomarkers, Tumor↗

Central projection of calcitonin gene-related peptide (CGRP)- and substance P (SP)-immunoreactive trigeminal primary neurons in the rat.

Substance P (SP) is implicated in transmission of primary afferent nociceptive signals. In primary neurons, SP is colocalized with calcitonin gene-related peptide (CGRP), which is another neuropeptide marker for small to medium primary neurons. CGRP coreleased with SP augments the postsynaptic effect of SP and thereby modulates the nociceptive transmission. This study demonstrates the distribution of CGRP-like immunoreactivity (-ir) and SP-ir in the lower brainstem of normal rats and after trigeminal rhizotomy or tractotomy at the level of subnucleus interpolaris (Vi). By comparing the results obtained from normal and deafferented rats, we analyzed the central projection of trigeminal primary nociceptors. The CGRP-immunoreactive (-ir) trigeminal primaries projected to the entire rostrocaudal extent of the spinal trigeminal nucleus, the principal nucleus (PrV), the paratrigeminal nucleus (paraV), and the lateral subnucleus of solitary tract nucleus (STN) on the ipsilateral side. The trigeminal primaries projecting to the spinal trigeminal nucleus, paraV and STN also contained SP-ir. The ipsilateral trigeminal primaries were the exclusive source of CGRP-ir terminals in the PrV, the Vi and the dorsomedial nucleus within the subnucleus oralis (Vo). The medullary dorsal horn (MDH) and the lateral edge of Vo received convergent CGRP-ir projection from the ipsilateral trigeminal primaries and other neurons. The glossopharyngeal and vagal primaries are candidates for the source of CGRP-ir projection to the Vo and the MDH, while the dorsal root axons supply the MDH with CGRP-ir terminals. In addition, contralateral primary neurons crossing the midline appear to contain CGRP and to terminate in the MDH.

Animals↗

S100 protein-immunoreactive primary sensory neurons in the trigeminal and dorsal root ganglia of the rat.

The cell body size (cross-sectional area) of S100-immunoreactive (-ir) primary neurons was measured in the trigeminal (TG) and lumbar dorsal root ganglia (DRG). About a half of neurons exhibited S100-immunoreactivity (-ir) in the DRG (44.0%) and TG (59.0%). DRG neurons with cell bodies > 1200 microm2 mostly exhibited S100-ir (96.5%), whereas S100-ir DRG neurons < 600 microm2 were rare (8.0%). 36.6% of DRG neurons in the cell size range 600-1200 microm2 showed the ir. TG neurons > 800 microm2 mostly exhibited S100-ir (93.1%), whereas those < 400 microm2 were devoid of it (positive cells 10.5%). 58.3% of TG cells in the range 400-800 microm2 contained S100-ir. Double-immunofluorescence method revealed the co-expression of S100 and other calcium-binding proteins. Parvalbumin-ir neurons mostly exhibited S100-ir in the DRG (97.4%) and TG (97.0%). The co-expression of S100 and calbindin D-28k was very rare in the DRG, because the DRG contained few calbindin D-28k-ir neurons. Unlike in the DRG, numerous neurons co-expressed S100- and calbindin D-28k-ir in the TG. Most calbindin D-28k-ir TG neurons were also immunoreactive for S100 (90.7%). Sub-populations of calretinin (CR)-ir neurons co-expressed S100-ir in both the DRG (68%) and TG (50.0%). Virtually all CR-ir neurons > 1400 microm2 co-expressed S100-ir in the DRG (100%) and TG (95.9%). CR-ir neurons < 800 microm2 were rarely exhibited S100-ir (DRG 18.0%, TG 21.9%). 71.3 and 60.5% of CR-ir neurons in the range 800-1400 microm2 co-expressed S100-ir in the DRG and TG, respectively. The present study indicates that S100 is closely correlated to the primary neuronal cell size in the DRG and TG.

Animals↗

Cloning and sequencing of trehalose synthase gene from Thermus aquaticus ATCC33923.

The gene encoding trehalose synthase (catalyzing the conversion of maltose into alpha, alpha-trehalose by transglucosylation) was cloned from Thermus aquaticus ATCC33923. Sequence analysis revealed a 2892 bp synthase gene and a 963 residue amino-acid sequence. The 547 N-terminal residues were homologous to the full-length synthase from Pimelobacter sp. R48 (53.8% identity).

Amino Acid Sequence↗

Deletion of the long arm of chromosome 20 in a patient with chronic neutrophilic leukemia: cytogenetic findings in chronic neutrophilic leukemia.

We encountered a 67-year-old female with chronic neutrophilic leukemia (CNL). Cytogenetic study showed she had a deletion in the long arm of chromosome 20. This finding indicates that CNL, in this case, is a clonal disorder. Most CNL patients have normal karyotypes, and only four patients with cytogenetic abnormalities, including two cases who received chemotherapy before the cytogenetic abnormality was detected, have been reported. Four of those cases, including our case, had abnormalities in the long arm of chromosome 20. This locus may be associated with the development of CNL. To our knowledge, this is the first case with CNL who showed deletion of the long arm of chromosome 20 before treatment was started.

Aged↗

Insulin-like growth factor-I mediates osteoclast-like cell formation stimulated by parathyroid hormone.

There have been several lines of evidence that parathyroid hormone (PTH) stimulates production of insulinlike growth factor I (IGF-I) in bone and that IGF-I stimulates osteoclast formation. Thus, the present study was performed to clarify the possible role of IGF-I in PTH-stimulated osteoclastlike cell formation and the role of PTH-responsive dual signal transduction systems (cyclic [c] AMP-dependent protein kinase [PKA] and calcium/protein kinase C [PKC]) in its mechanism. Treatment with anti-IGF-I antibody (1-10 micrograms/ml) partially but significantly blocked hPTH-(1-34)-stimulated osteoclastlike cell formation in unfractionated mouse bone cell cultures, although it did not affect osteoclastlike cell formation stimulated by 1,25-dihydroxyvitamin D3. Rp-cAMP5 (10(-4) M), a direct PKA inhibitor, as well as two types of PKC inhibitors, H-7 (10 microM) and staurosporine (3 nM), and dantrolene (10(-5) M), an inhibitor of calcium mobilization from intracellular calcium stores, all significantly blocked PTH-stimulated osteoclastlike cell formation. Anti-IGF-I antibody (3 micrograms/ml) significantly blocked osteoclastlike cell formation stimulated by 10(-4) M dbcAMP, 10(-4) M Sp-cAMPS, a direct PKA activator, and 10(-5) M forskolin in mouse bone cell cultures. Dibutyryl cAMP, forskolin, and hPTH-(1-34) significantly stimulated mRNA expression of both IGF-I and IGF-binding protein 5 (IGFBP-5) in these cultures, but neither 10(-7) M PMA, a PKC activator, nor 10(-7) M A23187 did. Moreover, anti-IGF-I antibody significantly blocked osteoclastlike cell formation stimulated by the conditioned medium from MC3T3-E1 cells pretreated with 10(-8) PTH-(1-34), which induced IGF-I and IGFBP-5 mRNA expression in these cells. In conclusion, the present study indicates that IGF-I mediates osteoclastlike cell formation stimulated by PTH and that the PKA pathway is involved in its mechanism. However, IGF-I does not seem to be the sole effector molecule to be active in this system.

Animals↗

Geriatric prescribing in the medical wards of a teaching hospital in Nepal.

A retrospective study on prescribing patterns for 100 randomly selected geriatric patients admitted over a period of 1 year to the medical wards of the Tribhuvan University Teaching Hospital (TUTH) in Nepal showed that polypharmacy was prevalent. During a hospital stay, 73% patients received more than five, 54% received more than eight, and 24% received more than nine drugs concurrently. Although the average drug exposure per patient during a hospital stay was found to be 11.2, this figure would actually go up to 14.5 if all the active ingredients of the fixed-dose combination products prescribed (15.4% of all drugs) were taken into account. Intravenous fluids were the most commonly prescribed drugs and were given to 91% of the patients. Antibiotics (excluding metronidazole and antituberculous drugs) were given to over three-quarters (77%) of the patients. Ciprofloxacin was the most commonly prescribed antibiotic. Of the 42 patients treated with this drug, 31 (73.8%) received it intravenously, either for a part of or throughout the course. This antibiotic was prescribed concurrently with theophylline in 14 patients without the facility for monitoring plasma-theophylline levels. It was also administered at the same time as antacid in nine patients. Nearly half (46.4%) of the drugs were prescribed by brand or proprietary names. The prescribing error of leaving the prescription card undated or unsigned when prescribing or stopping drugs was found to be high. The results of this survey indicate that there is considerable scope for improving geriatric prescribing practices in the medical wards of TUTH.

Journal Article↗

Parvalbumin- and calbindin D-28k-immunoreactive innervation of orofacial tissues in the rat.

Parvalbumin- and calbindin D-28k-immunoreactive (-ir) innervation was examined in orofacial tissues of the rat. Labial and facial skins were devoid of the calcium-binding protein (CaBP)-ir nerve endings, while the infraorbital and mental nerves contained numerous parvalbumin-ir axons. Labial and gingival mucosae were also devoid of the CaBP-ir nerve endings. The buccal mucosa and incisive papilla contained both encapsulated and unencapsulated endings, while the hard palate mucosa excluding the incisive papilla contained only unencapsulated endings. Encapsulated endings were found just beneath the epithelium or attached to the cartilaginous core of the incisive papilla. Unencapsulated endings in the lamina propria were subdivided into two types: simple (unramified) and complex (ramified). Neurites of simple endings were straight, curved, or coiled, while those of complex endings exhibited a bush-like appearance due to the ramification. In addition, palatal rugae contained intraepithelial endings. The unencapsulated complex endings in palatal rugae coexpressed parvalbumin- and calbindin D-28k-irs, whereas other endings were immunoreactive for parvalbumin alone. The pterygopalatine ganglion contained calbindin D-28k-ir pericellular fibers but not the ir cell bodies. A subpopulation of trigeminal ganglion neurons coexpressed both CaBPs. CaBP-ir encapsulated and unencapsulated endings in the oral mucosa probably include low-threshold mechanoreceptors, while parvalbumin-ir intraepithelial endings in the palatal mucosa may be involved in nociception.

Animals↗

Efficient in vivo gene transfer into the heart in the rat myocardial infarction model using the HVJ (Hemagglutinating Virus of Japan)--liposome method.

The lack of efficient treatment for myocardial infarction remains an unresolved problem in the field of cardiovascular disease. Gene therapy may be a potential therapeutic strategy for the treatment of myocardial infarction. However, current methods of in vivo gene transfer into the heart are limited by their low efficiency and/or potential toxicity. In the present study, we developed an efficient technique of gene transfer into the intact heart in vivo using the Sendai virus (HVJ: Hemagglutinating Virus of Japan)--liposome method. We used the beta-galactosidase gene, luciferase gene and human angiotensin converting enzyme (ACE) gene as markers. In vivo gene transfer into the rat heart was performed as follows: (1) direct injection into the rat heart, (2) incubation within the pericardium, and (3) infusion into a coronary artery. Direct injection of the HVJ-liposome complex containing the beta-galactosidase vector into the rat heart resulted in limited staining of beta-galactosidase 3 days after transfection. To compare transfection efficiency between "naked" plasmid DNA transfection and the HVJ-liposome method, we also transfected the luciferase reporter gene into the heart. Luciferase activity was significantly higher in hearts transfected by the HVJ-liposome method than that in hearts transfected by direct "naked" plasmid transfection (P < 0.01). To confirm the successful gene in the protein level, we measured ACE activity in the hearts. Cardiac ACE activity was significantly increased in hearts transfected with human ACE gene as compared to hearts transfected with control vector (P < 0.01). On the other hand, incubation of HVJ-liposome complex, containing beta-galactosidase vector, within the pericardium resulted in widespread staining of cardiac myocytes and fibroblasts, mainly located in several surface layers beneath the pericardium. More importantly, widespread stained areas of beta-galactosidase were also observed in the middle of the myocardium around the vasa vasorum. We also examined the efficiency of gene transfer by the HVJ-liposome method in a rat myocardial infarction model. In the infarction model, using the pericardium incubation approach, staining for beta-galactosidase was observed in the viable cells around the infarction area. Finally, direct infusion of the HVJ complex, containing the beta-galactosidase vector, into coronary artery also resulted in widespread staining of beta-galactosidase in cardiac myocytes around the microvasculature. Using direct injection, we found significant injury to the myocardium and severe fibrosis at the injection site, whereas no apparent injury was observed using pericardium incubation and coronary infusion. There was no evidence of cytotoxicity or inflammation caused by the HVJ-liposome complex itself. Overall, we have established an efficient in vivo gene transfer method into the heart using the HVJ-liposome method. Direct infusion into the coronary artery resulted in widespread transfection without damaging the myocytes; incubation within the pericardium demonstrated the usefulness of the HVJ-liposome method for studying cardiac function and as a means of gene therapy for cardiovascular diseases.

Animals↗

Neurogenic potential of Ewing's sarcoma cells.

We investigated the capacity of eight well-characterized Ewing's sarcoma cell lines to differentiate towards a neural phenotype. Ewing's sarcoma cells expressed the neuroectoderm-associated antigens such as nerve growth factor (NGF) receptor, N-CAM (6H7 and Leu-19) and Leu-7. Ewing's sarcoma cells also exhibited the potential for neural differentiation at the mRNA level; neuron-specific medium- and low-sized filament (NF-M and NF-L) expression was induced by dibutyryladenosine cyclic monophosphate. The pattern of expression of NF-L obtained by using alternative polyadenylation sites in Ewing's sarcoma cells differed from that in peripheral primitive neuroectodermal tumour (PNET) cells, and was similar to that in undifferentiated neural tissues. Furthermore, the NGF receptors detected by immunohistochemistry were found to be non-functional as assayed by c-fos induction with NGF treatment. The results indicate that Ewing's sarcoma cells maintain a primitive phenotype and have the potential to differentiate into a neural phenotype, indicating that Ewing's sarcoma is distinct from PNET.

Bone Neoplasms↗

Thyroid cancer in children: report of three cases and a review of the Japanese literature.

We experienced three cases of thyroid cancer in children less than 15 years of age between 1982 and 1995. We herein present these three cases with a review of 141 reported cases of childhood thyroid cancer in Japan. Our patients were 6, 13, and 14 years old. The patients, all girls, were diagnosed as having thyroid cancer based on diagnostic imaging. One of them was also diagnosed by a fine-needle aspiration biopsy (FNAB). One of them underwent subtotal thyroidectomy, and the other two underwent lobectomy. Modified neck dissections were performed on all three. Pathologically, the tumors were all papillary carcinomas. Multiple lymph node metastases were present in all patients. However, the postoperative courses have been good, and there have been no signs of recurrence, 10, 8, and 2 years after their respective operations. In 144 reported cases of childhood thyroid cancer in Japan including ours, the youngest patient was a 2-year-old boy, and the female to male ratio was 2.1:1. FNAB was performed in 25 cases, and 23 (92%) of the tumors were diagnosed as malignant. Histologically, 76% were papillary carcinoma and 20% follicular carcinoma. At operation, lymph node metastases were found in 80% of the cases, and lung metastases in 17%. For treatment, 88% of the patients received a more extensive operation than a lobectomy. Of the 144 patients, 8 died.

Adolescent↗

Epithelioid sarcoma producing granulocyte colony-stimulating factor.

An epithelioid sarcoma of the perineum of a 60-year-old man with widespread metastases produced leukocytosis, myeloid hyperplasia of the bone marrow, and splenomegaly. High titers of granulocyte colony-stimulating factor (G-CSF) were found in the patient's serum and primary culture medium of the tumor tissue. The tumor tissue extract contained m-RNA for G-CSF in large quantities, proving that the tumor was the source of this cytokine.

Granulocyte Colony-Stimulating Factor↗