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Biomedical subjects

Y Maruoka

Publications and source records attributed to Y Maruoka.

At least 19 recordsLinked to original sources

Bone with a vascular flap induced from fat tissue with the use of rhBMP-2 in rats.

Here we report that successful bone formation with a vascular flap inside a cylindrical mold was induced from fat tissue with the use of recombinant human bone morphogenetic protein-2 in rats. Fat tissue connected to blood vessels was prepared to fit into the mold and implanted intramuscularly into the hind leg in Wistar rats. RhBMP-2 (20 micro g) was applied in a collagen sheet previously placed on the inside surface of the mold. Bone formation was confirmed radiologically and morphologically at 2, 4, and 8 weeks after the surgery. In the control group without rhBMP-2 or the group with ligation of the blood vessels before the implantation, bone formation was not observed. Our success in bone formation having a definite size, shape, and blood supply may lead to a therapeutic approach to effective bone reconstitution. The present study is the first report on bone induction from fat tissue by rhBMP-2 in vivo.

Adipocytes↗

Blocking the glycine-binding site of NMDA receptors prevents the progression of ischemic pathology induced by bilateral carotid artery occlusion in spontaneously hypertensive rats.

This study was designed to investigate the participation of N-methyl-D-aspartate (NMDA) receptors in the progression of the pathology induced by bilateral carotid artery occlusion (BCAo) in spontaneously hypertensive rats (SHRs). We examined the effects of the selective NMDA receptor glycine-binding site antagonist SM-18400 on the mortality rate, deterioration of neurological signs, and formation of brain edema in the SHR-BCAo model. SM-18400 (15 or 30 mg/kg) was administered via the tail vein immediately and 2 h after BCAo. Neurological signs were monitored continuously for 8 h after BCAo, and the mortality rates were followed for 5 days. All SM-18400-treated animals were still alive 5 h after BCAo, whereas 38% of the animals died in the vehicle-treated group. The mortality rates of the SM-18400-treated groups were still lower than those of the vehicle-treated group 5 days after BCAo. In addition, SM-18400 markedly prevented the deterioration of neurological signs. The water content of the telencephalon and diencephalon/mesencephalon in the vehicle-treated group, measured 3 h after BCAo, was significantly higher than in the sham-operated group. SM-18400 significantly inhibited the increase in water content in both regions in a dose-dependent manner. These findings suggest that NMDA receptors participate in the increase in the mortality rate, deterioration of neurological signs, and formation of brain edema following ischemic brain damage in the SHR-BCAo model, and that SM-18400 can prevent ischemic insults.

Animals↗

Fusion variations of pancreatic ducts in patients with anomalous arrangement of pancreaticobiliary ductal system.

Fusion variations of the pancreatic ducts were studied to elucidate the significance of such variations. We classified structural fusion anomalies of the main and accessory pancreatic ducts on endoscopic retrograde cholangio-pancreatography (ERCP) in 37 patients with anomalous arrangement of the pancreaticobiliary ductal system (AAPB). The fusion variations of the pancreatic ducts were classified into five types: common, ansa pancreatica, branch fusion, looped, and separated. These fusion variations, except for common type, were found in 68% of the 37 patients with AAPB on ERCP. Fusion variations of the pancreatic ducts were very frequent (93%) in the 30 patients with congenital cystic dilatation of the common bile duct (CCD). The branch confluence fashion, in which the terminal bile duct communicated with a pancreatic duct branch, was found only in patients with cystic dilatation cyst of the CCD, and it appeared that cystic dilatation cyst of CCD might differ from spindle or cylindrical cyst originating from embryonic formation of an anomalous confluence. It was also suggested that in patients with fusion variations of the pancreatic ducts, the flow of pancreatic juice might be disordered, leading to the development of acute pancreatitis or pancreatic dysfunction. Consequently, it appears to be necessary to carefully examine patients with AAPB for the presence or absence of any fusion variations of the pancreatic ducts and to observe such patients with long-term monitoring by ERCP, and computed temography, and with pancreatic function tests.

Adult↗

Comparison of the expression of three highly related genes, Fgf8, Fgf17 and Fgf18, in the mouse embryo.

In mammals, 16 members of the Fgf family have so far been described with diverse roles in embryonic cell growth and differentiation. Here, we report the expression from early streak stage to midgestation of two newly-identified murine genes, Fgf17 and Fgf18, that are most closely related to Fgf8 (63.7% and 56.8% identical, respectively, at the amino acid level). Fgf17 is expressed during gastrulation but at lower levels than Fgf8, while Fgf18 RNA is not expressed until later, in paraxial mesoderm. In the developing tail bud, each Fgf gene shows a different pattern of transcription. Distinct and overlapping expression patterns are also described in the developing brain and limbs.

Animals↗

Overexpression of bcl-2 protein inhibits terminal differentiation of oral keratinocytes in vitro.

The bcl-2 proto-oncogene is a known inhibitor of apoptosis; in normal human stratified squamous epithelium, its expression is restricted to the basal cell layer. To investigate the functional role of bcl-2 protein in the process of differentiation of oral keratinocytes, bcl-2 expression vector was transfected into SCC-25 cells, which normally undergo squamous cell differentiation in vitro while expressing specific differentiation markers, e.g., keratin 10/11 and involucrin. In bcl-2 transfected SCC-25 cells, the expression of these differentiation markers was markedly suppressed. The bcl-2 proto-oncogene may play a critical role in opposing the commitment to terminal differentiation and apoptosis of oral keratinocytes.

Apoptosis↗

Effects of the novel tricyclic quinoxalinedione derivatives, SM-18400 and its analogs, on N-methyl-D-aspartate (NMDA) receptor-mediated synaptic transmission in the isolated neonatal rat spinal cord in vitro.

We examined the effects of novel tricyclic quinoxalinedione derivatives, SM-18400 ((S)-9-chloro-5-[p-aminomethyl-o-(carboxymethoxy)phenylcarbamoylmethy l]-6,7-dihydro-1H,5H-pyrido[1,2,3-de]quinoxaline-2,3-dione hydrochloride trihydrate) and its analogs (i.e., ID-17263 and ID-17332), on the N-methyl-D-aspartate (NMDA) receptor-mediated polysynaptic reflex (PSR) in the isolated spinal cord of neonatal rats in vitro. Application of SM-18400 selectively suppressed the PSR activity in a concentration-dependent manner without affecting the monosynaptic reflex (MSR). Differential suppression of the PSR was also obtained with ID-17263, ID-17332 and other known NMDA receptor glycine-binding site antagonists, 5,7-dichlorokynurenate (5,7-diClkyn) and L-689,560 (4-trans-2-carboxy-5,7-dichloro-4-phenylaminocarbonylamino-1,2,3,4 -tetrahydroquinoline). Relative potencies of the test drugs for inhibition of the PSR were as follows: SM-18400 >> L-689,560 > ID-17332 > ID-17263 > 5,7-diClkyn. In addition, the inhibitory effects of SM-18400 on PSR were markedly antagonized by simultaneous application of D-serine, an agonist for NMDA receptor glycine-binding sites. These findings suggest that SM-18400 is a potent NMDA receptor glycine-binding site antagonist and blocks the NMDA receptor-mediated synaptic neurotransmission in the spinal cord in vitro.

Aminoquinolines↗

Keratinocytes become terminally differentiated in a process involving programmed cell death.

Oral keratinocytes originate from basal cells, differentiate during migration to the surface, and finally are shed. Apoptosis occurs at the end of differentiation, but the precise relationship between terminal differentiation and apoptosis is not clear. In the present study, Bcl-xL was expressed in the basal cell and spinous cell layers, and Bax was expressed in the spinous cell and granular cell layers. In cultured keratinocytes, Bcl-xL was expressed under conditions of 0.1 mM calcium (low Ca2+) but disappeared under conditions of 1.0 mM calcium (high Ca2+); the latter induces keratinocyte differentiation. Bax was not expressed in keratinocytes with low Ca2+ but was expressed in cells with high Ca2+. Finally keratinocytes with high Ca2+ underwent apoptosis, which was detected by the TUNEL method and by 180-bp DNA fragmentation. These results suggest that the process of terminal differentiation in gingival epithelium is a pathway to apoptosis.

Apoptosis↗

Selective depression of the spinal polysynaptic reflex by the NMDA receptor antagonists in an isolated spinal cord in vitro.

1. The effects of N-methyl-D-aspartate (NMDA) receptor glycine-binding site antagonists 7-chlorokynurenate (7-Clkyn) and (+/-)-3-amino-1-hydroxy-2-pyrrolidone (HA-966) on spinal reflexes in an isolated spinal cord that was maintained in Mg(2+)-free medium in vitro were examined. The actions of 7-Clkyn and HA-966 were compared with those of the channel-site antagonist (i.e., dizocilpine) and NMDA-binding site antagonists--that is, 3-[(+/-)-2-carboxypiperazin-4-yl]-propyl-1-phosphonate (CPP) and DL-2-amino-5-phosphonovalerate (APV). 2. 7-Clkyn and HA-966 produced a selective depression of the polysynaptic reflex (PSR) while negligibly affecting the activity of the monosynaptic reflex (MSR). The PSR was also differentially suppressed by dizocilpine, CPP and APV. The PSR inhibitory activity of the NMDA antagonists was in the following order: dizocilpine > CPP > APV = 7-Clkyn > HA-966. 3. The inhibitory effects of 7-Clkyn on PSR were markedly antagonized by the simultaneous application of D-serine, an agonist for the NMDA receptor glycine-binding sites. However, PSR inhibition by dizocilpine and CPP was unaffected. 4. Inhibition of the PSR by 7-Clkyn persisted in the presence of strychnine, which markedly increased the PSR activity by itself. 5. These findings suggest that the NMDA receptor glycine-binding sites play a role in generating the NMDA receptor-mediated PSR in the spinal cord in vitro.

Animals↗

Expression of mRNA encoding intestinal type alkaline phosphatase in rat liver and its increase by fat-feeding.

The presence of types of alkaline phosphatase (ALP) other than the tissue non-specific type enzyme in rat liver and its increase by fat feeding are known. In order to examine expression of intestinal type ALP in liver, specific oligonucleotide primers corresponding to two types of mRNAs of rat intestinal ALP (RTIN-1 and -2) were designed and amplified by means of the reverse transcriptase-polymerase chain reaction (RT-PCR). It was found that RTIN-1 mRNA was expressed only in the intestine but not in the liver, while RTIN-2 mRNA was expressed both in the intestine and in the liver. By fat feeding, expression of RTIN-1 mRNA increased in the intestine and that of RTIN-2 mRNA increased both in the intestine and in the liver. Thus, it was concluded that rat liver expressed one of the intestinal type ALP (RTIN-2) which was enhanced by fat feeding.

Alkaline Phosphatase↗

Blood pressure response during dental surgery.

To investigate blood pressure and pulse rate responses to dental surgery, 21 patients 18 to 73 years of age (mean age, 42 +/- 4 years) who visited our hospital for tooth extraction were studied. Before dental treatment, the patients underwent a mental arithmetic stress test, electrocardiography, and an anxiety evaluation with the State-Trait Anxiety Inventory. Baseline blood pressure and pulse rate were 118 +/- 4/70 +/- 3 mmHg and 69 +/- 2 beats/min, respectively. Blood pressure rose by 24 +/- 3/17 +/- 2 mmHg during the mental stress test, and the magnitude of the rise in systolic blood pressure was significantly correlated with age (r = 0.81, p < 0.001) and baseline blood pressure (r = 0.56, p < 0.01). After the topical injection of local anesthetic containing 1: 80,000 epinephrine, a transient increase in systolic blood pressure was observed. The maximum blood pressure and pulse rate increases during dental surgery were 24 +/- 4/13 +/- 2 mmHg and 17 +/- 3 beats/min, respectively. Similarly, the rate pressure product increased from 8,196 +/- 486 to 11,802 +/- 682. The magnitude of the blood pressure increase during dental surgery was not correlated with age, sex, family history of hypertension, baseline blood pressure, anxiety score, or response to mental stress. On the other hand, when the subjects were divided into two subgroups according to the blood pressure response during dental surgery, the larger response group (increase in mean blood pressure greater than 15 mmHg, n = 9) required a significantly larger dose of local anesthetic than did the smaller response group. The number of cases of pericoronitis of the third molar tended to be greater in the larger response group. These results indicate that an increase in blood pressure during dental surgery cannot be predicted on the basis of baseline blood pressure or the response to mental stress, but is related to the cause of tooth extraction and the volume of local anesthetics required to control the pain.

Adolescent↗

Identification of bone-type alkaline phosphatase mRNA from human periodontal ligament cells.

Tissue-nonspecific-type alkaline phosphatase is found in the bone, liver, kidney, and other tissues, and its gene consists of 12 exons with the coding sequence beginning in the second exon. Recently, an alternative noncoding first exon was identified in the liver message which differed from that of the previously known osteoblast-derived cDNA sequence. Although these two mRNAs produce an identical protein, they have different promoter regions. The periodontal ligament tissue expresses a high level of alkaline phosphatase activity. To identify its mRNA type, we isolated a full-length cDNA for alkaline phosphatase from a cultured human periodontal ligament cell expression library, using bone-derived tissue-nonspecific alkaline phosphatase cDNA as a hybridization probe. The size of this clone was 2.5 kb, and its 5' and 3' untranslated sequences were identical to those of the human tissue-nonspecific type isolated from osteoblastic cells but not to those of the liver type. In addition, the same fragments as in bone-derived tissue-nonspecific-type cDNA were detected by the treatment of the cDNA clone with restriction enzymes Hinc II and Pst I. The results suggest that expression of the same alkaline phosphatase isozyme in human periodontal ligament cells may be regulated by the same transcriptional mechanism as in bone.

Alkaline Phosphatase↗

Molecular cloning and sequence of bovine Msx-1 homeobox-containing gene cDNA from a bovine odontoblast library.

Screening of a bovine odontoblast cDNA library from developing incisor with murine Msx-1 and Msx-2 cDNA probes led to the isolation of three positive clones. All of them encoded for a sequence of a protein containing 297 amino acids. The responsible gene was designated as bovine Msx-1 (bMsx-1) due to the high homology with the human MSX-1 and mouse Msx-1 sequences.

Amino Acid Sequence↗

Cloning and sequence of bone morphogenetic protein 4 (BMP-4) from a human placental cDNA library.

A cDNA clone encoding bone morphogenetic protein-4 (BMP-4) has been isolated from a human placental cDNA library. Sequence analysis of this clone revealed that the nucleotide sequence of 5' region was different from that of human osteosarcoma BMP-4 and the deduced amino acid sequence indicated deletion of N-terminal 6 amino acids. We confirmed the expression of this type of BMP-4 mRNA in one human osteogenic cell line in addition to the placenta by the polymerase chain reaction (PCR).

Amino Acid Sequence↗

Production of functional human bone morphogenetic protein-2 using a baculovirus/Sf-9 insect cell system.

A clone of a human Bone Morphogenetic Protein-2 (hBMP-2) cDNA was obtained from a cDNA library established from human dental pulp cells. After subcloning hBMP-2 cDNA into Autographa californica nuclear polyhedrosis virus, the recombinant baculovirus was transfected to Sf-9 cells. Immuno-reactive recombinant hBMP-2 (rhBMP-2) was detected by a polyclonal antibody against Xenopus BMP-2 in the transfected insect cells but not in the culture media. Three days after treatment with the lysate of the transfected Sf-9 cells, increase in alkaline phosphatase activity of a murine stromal cell line, ST2, was detected. Subcutaneous implantation of rhBMP-2 produced in the insect cells induced formation of cartilage, bone and bone marrow in the rats. The present data indicated that the rhBMP-2 preparation produced in the insect Sf-9 cells had a comparable activity to that produced in mammalian cells.

Alkaline Phosphatase↗

Modulation of responses to TGF-beta by 1, 25 dihydroxyvitamin D3 in MG-63 osteoblastic cells: possible involvement of regulation of TGF-beta type II receptor.

The effect of TGF-beta 1 on the expression of alkaline phosphatase (ALPase) activity was examined during osteoblastic cell line (MG-63) differentiation induced by 1,25-dihydroxyvitamin D3 (1,25D3). TGF-beta 1 and 1,25D3 were found to enhance ALPase activity. However, preincubation of the cells with 1,25D3 transiently abolished the effects of TGF- beta 1. Kinetics of the complex responses to TGF- beta 1 and 1,25D3 were found to correlate well with that of the expression level of type II receptor for TGF- beta. These results suggest that 1,25D3 may regulate the cellular responses to TGF- beta 1 in part via regulation of functional receptor for TGF- beta.

Alkaline Phosphatase↗