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Biomedical subjects

T Sugimoto

Publications and source records attributed to T Sugimoto.

At least 19 recordsLinked to original sources
PubMed1995

EWS-FLI-1 and EWS-ERG chimeric mRNAs in Ewing's sarcoma and primitive neuroectodermal tumor.

The t(11;22)(q24;q12) and t(21;22)(q22;q12) are specific chromosomal translocations found in the Ewing family of tumors including ES, PNET and Askin tumors. In these translocations, the amino-terminal portion of the EWS gene located in 22q12 fuses to the carboxyl-terminal portion of the FLI-1 gene located in 11q24 or the ERG gene located in 21q22, which belong to the ets oncogene superfamily of transcription activators. We investigated the chimeric mRNAs of 15 ESs (7 cell lines and 8 tumor samples) and 7 PNETs (3 cell lines and 4 tumor samples) using the RT-PCR method and sequencing. We detected 2 types of EWS-ERG chimeric mRNA in 2 ES cell lines and 1 PNET tumor sample in addition to 4 types of EWS-FLI-1 chimeric mRNA in 11 ESs (4 cell lines and 7 tumor samples) and 4 PNETs (2 cell lines and 2 tumor samples). There seemed to be no association between the type of chimeric mRNA and clinical features such as sex, age, primary site and histopathology of the patients. All of the chimeric mRNAs are generated from in-frame junctions and are thought to encode fusion proteins that may be the molecular mechanism involved in the Ewing family of tumors.

Adolescent↗
PubMed1995

Molecular characterization of cDNA encoding a novel protein related to transcriptional enhancer factor-1 from neural precursor cells.

We identified a novel cDNA related to that of transcriptional enhancer factor-1 (TEF-1) during the course of isolation and characterization of cDNAs, whose mRNAs are preferentially expressed in the mouse neural precursor cells. The putative polypeptide, termed embryonic TEA domain-containing factor (ETF), deduced from the nucleotide sequence contains 445 amino acids and shares 66% amino acid identity with mouse and human TEF-1 proteins. The primary structure of the TEA domain, a probable DNA-binding domain, and the specific DNA binding activity to the GT-IIC motif of ETF are indistinguishable from those of the known vertebrate TEF-1 proteins. However, the expression of the ETF gene is strictly regulated in developing embryos and is limited to certain tissues, such as the hindbrain of a 10-day-old mouse embryo, in contrast to the ubiquitous expression pattern of the TEF-1 gene. These results suggest that ETF is a novel mammalian member of the TEA domain-containing transcription factor family and may be involved in the gene regulation of the neural development. We have discussed the possible existence of multiple subtypes of the mammalian TEF-1 family proteins, which may play different roles in cellular and development gene regulation.

Amino Acid Sequence↗
PubMed1995

Molecular cloning and characterization of rat V1b vasopressin receptor: evidence for its expression in extra-pituitary tissues.

Arginine vasopressin (AVP) modulates the secretion of ACTH through vasopressin receptor subtype V1b in the pituitary. We recently cloned human V1b, but several inconsistencies were found between the characteristics of this cloned receptor and those of rat pituitary membrane shown by previous workers. To clarify this issue, we report here the molecular cloning and functional expression of the cDNA encoding V1b in rat pituitary. This receptor encodes 425 amino acid protein having the highest identity with the human V1b (81%). Expression of this receptor in COS-1 cells showed pharmacological characteristics of V1b consistent with those of rat pituitary membrane. Northern blot and RT-PCR analyses revealed that mRNA of this receptor was expressed not only in anterior pituitary, but also in other tissues. This finding raises the possibility that V1b may play a role in regulating cell functions of these tissues.

Amino Acid Sequence↗
PubMed1995

The structure of the Aplysia kurodai gene encoding ADP-ribosyl cyclase, a second-messenger enzyme.

The complete nucleotide (nt) sequences of the cDNA and gene encoding the marine mollusk Aplysia kurodai (Ak) ADP-ribosyl cyclase (ADRC) which synthesizes cyclic ADP-ribose (cADP-ribose), a second messenger for Ca2+ mobilization from endoplasmic reticulum, were determined. Ak ADRC consists of 258 amino acids (aa) (29 kDa). It shares 86% aa sequence homology with that from A. californica, and 31-32% homology with the human, rat and mouse cluster of differentiation 38 (CD38) that has both ADRC and cADP-ribose hydrolase activities. The Ak ADRC-encoding gene (ADRC) spans approx. 7 kb and contains eight exons and seven introns. The transcription start point (tsp) determined by primer extension analysis and S1 mapping is 28 bp downstream from the TATA box. This gene is expressed specifically in the ovotestis, although the mammalian CD38-encoding gene is expressed in many kinds of tissues and cells. The 5'-flanking region contains several consensus sequences responsible for the germ-cell-specific expression of the mouse zona pellucida 3 (ZP3) and Drosophila melanogaster chorion genes. The existence of the consensus sequences located at nt -1649, -1161, -234 and -90 may account for the ovotestis-specific expression of the Ak ADRC gene.

ADP-ribosyl Cyclase↗
PubMed1995

Parvalbumin- and calretinin-immunoreactive trigeminal neurons innervating the rat molar tooth pulp.

Calcium-binding proteins and neuropeptides were examined in trigeminal neuronal cell bodies retrogradely labeled with Fast blue (FB) from the maxillary molar tooth pulp of the rat. FB-labeled cells were located in the maxillary division of the trigeminal ganglion. Approximately 30 and 50% of the labeled cells were immunoreactive for parvalbumin and calcitonin gene-related peptide (CGRP), respectively. The coexpression of these substances was observed in 9.5% of FB-labeled cells. On the other hand, 2.4% of FB-labeled cells exhibited calretinin-immunoreactivity (CR-ir) and 20% tachykinin (TK)-ir. The coexpression of CR and TK was observed in 1.9% of FB-labeled cells, i.e., most of CR-ir FB-labeled neurons coexpressed TK-ir. An immuno-EM method revealed that all parvalbumin-ir nerve fibers in the root pulp were myelinated and that CGRP-ir nerve fibers were both myelinated (15%) and unmyelinated (85%). The present study indicated that primary nociceptors innervating the rat molar tooth pulp contained parvalbumin and CR and coexpressed these calcium-binding proteins and neuropeptides. It was suggested that peripheral axons of parvalbumin-ir tooth pulp primary neurons are all myelinated. Most peripheral CR-ir axons are probably unmyelinated because TK-ir myelinated axons have never been demonstrated in any peripheral organ.

Animals↗
PubMed1995

CGRP in peripherally axotomized mesencephalic trigeminal neurones of the rat.

Immunoreactivities for substance P (SP-ir) and calcitonin gene-related peptide (CGRP-ir) were examined in neurones in the mesencephalic trigeminal tract nucleus of nerve-injured rats. One week following unilateral transection of the trigeminal root, the masseteric nerve (MN) or the inferior alveolar nerve (IAN), CGRP-ir but not SP-ir was detected in these neurones bilaterally with ipsilateral dominance. Most of the CGRP-ir (immunoreactive) neurones were retrogradely labelled when fluorogold (FG) had been applied to the proximal stump of the MN or the IAN. Sham surgery including insult to the masseter muscle induced CGRP-ir while skin incision did not.

Animals↗
PubMed1995

Plasma adenosine concentrations are elevated in Dahl salt-sensitive rats.

We measured plasma levels of adenosine in Dahl salt-sensitive rats (DS) and Dahl salt-resistant rats (DR) to examine the potential role of adenosine in cardiovascular regulation in this type of hypertension. Plasma adenosine concentrations were significantly higher in DS than in DR. The NaCl content in the diet did not affect plasma adenosine concentration in either DS or DR. Significant positive correlation was found between adenosine concentrations and systolic blood pressure when the data for DS and DR were analyzed together. These results suggest that adenosine may play an important role in the pathophysiology of hypertension in DS.

Adenosine↗
PubMed1995

Determination of urinary valproylcarnitine by gas chromatography-mass spectrometry with selected-ion monitoring.

A modified method for the determination of valproylcarnitine in urine samples of patients receiving sodium valproate by gas chromatography-mass spectrometry with selected-ion monitoring is described. The chemically analogous internal standard 2-ethylpentanoylcarnitine was added to the urine samples. Valproic acid and its metabolites were removed by extraction with chloroform at pH 5.0. The samples were then applied onto a C18 Sep-Pak column. Inorganic and water soluble compounds were washed out with water. Valproylcarnitine and internal standard were eluted with methanol and were derivatized to the corresponding acyl-containing lactones by heating at 100 degrees C for 60 min in dimethylformamide. Urinary valproylcarnitine levels of epileptic patients receiving valproate were determined according to the present method. The data obtained might be useful for diagnosis of carnitine deficiency.

Calibration↗
PubMed1995

Differences in body temperature changes during dressing change in surviving and non-surviving burned patients.

Change in body temperature (delta BT) while dressings were being changed was measured in 52 extensively burned adults. The patients were divided into four groups based on outcome and the prognostic burn index (PBI; age + burn index): group I (15 survivors, PBI less than or equal to 80), group II (15 survivors, PBI between 80 and 120), group III (14 non-survivors, PBI between 80 and 120), and group IV (eight non-survivors, PBI greater than 120). The body temperature before the change of dressings was the same in the four groups. The mean(s.d.) delta BT in all patients was -0.5(0.8) degrees C. The magnitude of delta BT was not related to the time required for changing dressings (r = 0.04) or to the non-epithelialized area present at each change of dressings (r = -0.05). All groups showed a similar increase in delta BT during the early period after the burn, but in group I it began to decrease progressively after the 16th day after the burn (P < 0.05). A reduction in delta BT was noted in group II from day 25, whereas it was unchanged in groups III and IV (P < 0.05). It also significantly increased 2 days before the diagnosis of sepsis (P < 0.05). These results emphasize that delta BT may provide useful information about prognosis and the development of sepsis.

Adult↗
PubMed1995

Adaptive cytoprotection in cultured rat gastric mucus-producing cells. Role of mucus and prostaglandin synthesis.

In cultured gastric mucosal cells, we investigated whether: (1) adaptive cytoprotection was associated with stimulation of endogenous prostaglandin synthesis; (2) prostaglandins given exogenously were cytoprotective against ethanol-induced gastric mucosal cell damage; and (3) a relationship existed between cytoprotection and mucus release. Cytolysis was quantified by measuring 51Cr release from prelabeled cells. Mucus release was determined by measurement of [3H]glucosamine release. Concentrations of ethanol > 12% caused cell damage and increased 51Cr release dose dependently. Pretreatment with low concentrations of ethanol (0.5-1.5%) decreased ethanol-induced 51Cr release, but also decreased prostaglandin E2 synthesis. Prostaglandin E2 and 16,16-dimethyl prostaglandin E2 given exogenously were cytoprotective against ethanol-induced gastric mucosal cell damage. Treatment with low concentrations of ethanol (1.5%) increased mucus release from cultured gastric mucosal cells. However, prostaglandin E2 and 16,16-dimethyl prostaglandin E2 did not affect mucus release. We conclude that in cultured gastric mucus-producing cells: (1) adaptive cytoprotection occurs without stimulation of endogenous prostaglandin synthesis but with increase in mucus release; and (2) exogenous prostaglandins are cytoprotective against ethanol-induced gastric mucosal cell damage without stimulating mucus release in vitro. We postulate that adaptive cytoprotection in cultured gastric mucus-producing cells is not mediated by prostaglandin, but by mucus released in response to a mild irritant.

16,16-Dimethylprostaglandin E2↗
PubMed1995

Retinoic acid induces osteoclast-like cell formation by directly acting on hemopoietic blast cells and stimulates osteopontin mRNA expression in isolated osteoclasts.

Although retinoic acid (RA) has been considered to be a bone-resorbing agent both in vivo and in vitro, its mechanism remains still unclear. The present study was performed to examine the effect of RA on osteoclast-like cell formation in the presence or absence of osteoblasts and to study whether RA would affect osteopontin mRNA expression in isolated rabbit osteoclasts. RA (10(-8) and 10(-6) M) significantly stimulated the formation of osteoclast-like cell in osteoblast-containing mouse bone cell cultures. Also, RA caused a stimulation of osteoclast-like cell formation from hemopoietic blast cells supported by granulocyte macrophage-colony stimulating factor (GM-CSF) in mouse spleen cell cultures. However, RA did not affect blast cell number in these cultures and significantly inhibited GM-CSF-stimulated proliferation of hemopoietic blast cells. On the other hand, RA stimulated the bone-resorbing activity of mature osteoclasts in mouse bone cell cultures. Moreover, RA caused a stimulation of osteopontin mRNA expression in isolated rabbit osteoclasts. The present study demonstrated for the first time that RA stimulated osteoclast-like cell formation, presumably through directly acting on the hemopoietic blast cells, and that RA stimulated osteopontin mRNA expression in isolated rabbit osteoclasts.

Animals↗
PubMed1995

A subpopulation of large ganglion neurons express IsK protein mRNA: an in situ hybridization analysis in the rat eye.

Rat IsK protein is a 130-amino acid membrane protein and subserves generation of K+ outward current. The expression of this putative K+ ion channel mRNA was explored in the eye with in situ hybridization. The ganglion cell layer of the retina expressed IsK mRNA in a subpopulation of ganglion cells composed of large cell bodies. The hybridization-positive cells were scattered throughout the retina. The hybridization signal was also localized to the epithelial cells throughout the cornea. The results provided evidence for IsK message in neuronal cells. Expression of IsK message in the eye was thus shown to be restricted to particular cell types.

Animals↗
PubMed1995

Successful treatment of hyperphosphatemic tumoral calcinosis with long-term acetazolamide.

We describe a patient with tumoral calcinosis, in which acetazolamide (ACZ) was, for the first time, tested for its therapeutic efficacy. The 19-year-old Japanese man had been suffering from multiple recurrent calcific masses with tenderness around the finger, knee, and toe joints since 10 months of age. Radiographs revealed several calcific subcutaneous masses around the finger joints, and calcific myelitis around the right knee joint and in the calvarium. The patient had hyperphosphatemia with elevated maximal threshold of renal phosphate excretion in the presence of normal kidney function and normocalcemia, suggesting a reduced ability to excrete phosphorus in the urine. A delay of disappearance of orally administered phosphate from the blood stream was found. A serum parathyroid hormone (PTH) level was normal, and responses to PTH and ACZ were also normal regarding the induction of phosphaturia. Since the masses tended to recur easily despite repeated surgical resections, we started medical treatment with phosphorus deprivation by oral aluminum hydroxide. However, the drug alone had no effect on hyperphosphatemia or calcific lesions, and ACZ was added in expectation of making the patient's phosphorus balance negative by its phosphaturic effect. Fourteen years of administration of the two drugs apparently improved the patient's symptoms, the biochemical findings, and the calcific lesions on radiographs. Thus, ACZ appeared to be useful for tumoral calcinosis resistant to phosphorus deprivation by aluminum hydroxide alone.

Acetazolamide↗
PubMed1995

Low oxygen extraction despite high oxygen delivery causes low oxygen consumption in patients with burns recovering slowly from operative hypothermia.

BACKGROUND: Patients with burns who eventually succumbed to their injuries tended to recover more slowly from operative hypothermia than those who survived. Slower recovery was associated with a lower postoperative oxygen consumption (VO2). We have now investigated whether this was due to impairment of oxygen delivery or extraction. METHODS: This study was performed in 13 adult patients with severely burns. One hundred four measurements of VO2 by indirect calorimetry were made during recovery from 23 episodes of operative hypothermia in 11 patients. Sixty-six measurements of oxygen transport variables by balloon-tipped pulmonary artery catheter were made after 17 episodes of operative hypothermia in six patients. Body temperature was monitored in the urinary bladder. RESULTS: The rate of temperature rise (T) showed a strong positive correlation with VO2 measured both by indirect calorimetry (r = 0.91, p < 0.001) and by balloon-tipped pulmonary artery catheter (r = 0.83, p < 0.001). Oxygen delivery (DO2) was above normal in nearly all patients. Oxygen extraction was low in patients recovering slowly (T < 1.0 degree C/hr) and high in those recovering quickly (T > or = 1.0 degree C/hr). During fast recovery VO2 (373 +/- 77 ml.min-1.m-2; mean +/- SD) was approximately three times normal and was independent of DO2. In contrast, a strong linear relationship existed between VO2 and DO2 during slow recovery (r = 0.76, p < 0.001). CONCLUSIONS: Patients with burns with slow recovery from operative hypothermia exhibited impaired oxygen extraction and dependence of VO2 on DO2 over a wide range. This picture resembles that in patients with critical illness.

Adult↗
PubMed1995

Hydrolysis of a carboxy-terminal fragment of parathyroid hormone-related protein by rat kidney: evidence for a crucial role of meprin.

Although parathyroid hormone-related protein (PTHrP) is known to be secreted into the circulation as heterogeneous forms, with its N-terminal and C-terminal fragments as well as the intact form, the fate of these molecules in the plasma has not been fully understood. As for a C-terminal fragment, the kidney seems to be physiologically important in its metabolism, because it is elevated in circulating plasma in patients with chronic renal failure. In this study, we examined the mechanism by which a C-terminal fragment of PTHrP was metabolized by rat kidney and by other rat organs in vitro. When human (h) PTHrP-(109-141) was incubated for 2 h with an extract of rat kidney, it was almost completely hydrolyzed. This hydrolysis was readily blocked by the additions of o-phenanthrolline and dithiothreitol, indicating the participation in this process of a metallo-protease possessing disulfide bonds. This hydrolytic activity showed a meprin-like character, with being sensitive to actinonin but not to phosphoramidon. Furthermore, the participation of meprin itself in this process was directly confirmed by the experiment comparing degradation products of the peptide by the microvillar membranes of rat kidney with those by a purified rat meprin, in which a large part of the metabolites by rat kidney membranes corresponded to those by rat meprin. Among other rat organs examined, the extracts of the small intestine pancreas, spleen and urinary bladder exerted remarkable hydrolytic activities for hPTHrP-(109-141).(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1995

Prolongation of intraventricular conduction time associated with fatal [correction of fetal] impairment of defibrillation efficiency during treatment with class I antiarrhythmic agents.

To test whether fatal deterioration of defibrillation efficiency during antiarrhythmic therapy can be prevented by avoiding extreme decrease in ventricular prevented by avoiding extreme decrease in ventricular conduction or toxic plasma drug levels, we determined the defibrillation threshold (DFT) before and during infusion of incremental doses of disopyramide (n = 8), mexiletine (n = 9), or flecainide (n = 9) in anesthetized dogs. Disopyramide did not alter DFT [from 4.4 +/- 1.5 to 4.4 +/- 1.6 J (3.1 +/- 1.2 micrograms/ml)]. Mexiletine tended to increase DFT [from 4.6 +/- 1.2 to 6.1 +/- 2.0 J (1.8 +/- 0.6 micrograms/ml); p < 0.05], and defibrillation eventually was unsuccessful in 3 of the 9 dogs. Although the plasma mexiletine level before refractory fibrillation was far beyond the human therapeutic range, prolongation of intraventricular conduction time (CT) was moderate (16 +/- 3%). Flecainide increased DFT from 4.2 +/- 1.3 to 6.1 +/- 1.5 J at a plasma level of 1.04 +/- 0.37 micrograms/ml (p < 0.0005). In 3 of 5 dogs that developed refractory fibrillation, plasma flecainide level before terminal ventricular fibrillation (VF) was not toxic, but prolongation of CT in the 5 dogs was remarkable (30 +/- 9%). Thus, VF resistant to defibrillation is not necessarily associated with both toxic plasma drug level and remarkably decreased conduction. Reliability of these valuables as indicators of fatally deteriorated defibrillation efficiency may vary among antiarrhythmic agents.

Analysis of Variance↗
PubMed1995

Relative contribution of alpha 1-adrenoceptor blocking activity to the hypotensive effect of the novel calcium antagonist monatepil.

Monatepil, a novel calcium antagonist, has alpha 1-adrenoceptor blocking activity; in the present study, we examined the relative contribution of this alpha 1-blocking activity to its hypotensive effect. Monatepil and diltiazem produced dose-dependent hypotensive effects in anesthetized rats with the same potency. Prazosin and monatepil inhibited the L-phenylephrine (L-PE)-induced pressor response, whereas diltiazem scarcely did. The injection of prazosin produced a decrease in blood pressure (BP) in anesthetized rats. The decrease was recovered with angiotensin II (AII) infusion in a dose-dependent manner. We developed a new rat model by first intravenously injecting prazosin and then infusing AII in anesthetized rats. In this model, diltiazem produced the same hypotensive effect as it did in pretreated conditions, although the hypotensive effect of monatepil was attenuated by 20-35% as compared with pretreated conditions. These results suggest that monatepil exerts alpha 1-adrenoceptor blocking action in vivo and 20-35% of the hypotensive effect of monatepil is attributed to its alpha 1-adrenoceptor blocking activity.

Adrenergic alpha-Antagonists↗