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Biomedical subjects

N Koyama

Publications and source records attributed to N Koyama.

At least 73 records · Page 4Linked to original sources

Differential activation of spinal dorsal horn units by subcutaneous formalin injection in the cat: an electrophysiological study.

In our previous report we found that subcutaneous (s.c.) formalin injection into the cutaneous receptive field (RF) of dorsal horn wide-dynamic-range (WDR) units and nociceptive primary afferent units resulted in a tonic, long-lasting increase in firing. However, s.c. formalin injection only resulted in a short-lasting increase in firing of non-nociceptive primary afferent units. In the present study, by using extracellular single-unit recording techniques we further studied effects of s.c. formalin on response properties of identified superficial-layer nociceptive-specific (NS) units and deeper-layer, low-threshold mechanoreceptive (LTM) units of L7 dorsal horn in urethane-chloralose-anesthetized cats. s.c. formalin injection into the RF of NS units resulted in a tonic, long-lasting increase in firing (7.08+/-0.42 spikes/s, n = 5), for more than 1 h, compared with the spontaneous background (1.42+/-0.03 spikes/s, n = 5). Formalin injection into the RF of LTM units also resulted in an increase in firing; however, the duration was short-lasting, for 25-520 s (152.92+/-46.73 s, n = 12). The present study demonstrated that s.c. injection of dilute formalin solution resulted in activation of not only nociceptive but also non-nociceptive dorsal horn units, suggesting that tissue injury caused by s.c. formalin results in vigorous injury discharges of peripheral nerve terminals, which subsequently leads to activation of primary afferent neurons and secondary dorsal horn neurons.

Animals↗

Possible involvement of cysteine and histidine residues in the (NH4+ + Na+)-activated ATPase of an anaerobic alkaliphile, Amphibacillus xylanus.

Effect of various inhibitors on the (NH4+ + Na+)-activated ATPase of an anaerobic alkaliphile, Ep01(a strain of Amphibacillus xylanus), was examined. Among the chemicals tested, the enzyme was drastically inactivated by p-chloromercuribenzoic acid and diethyl pyrocarbonate. The ATPase activity of the enzyme, which was inactivated by p-chloromercuribenzoic acid and diethyl pyrocarbonate, was remarkably restored by beta-mercaptoethanol and hydroxylamine, respectively, suggesting the involvement of cysteine and histidine residues in the enzyme activity. Analysis of the inhibition kinetics by diethyl pyrocarbonate indicated that modification of a single histidine residue per ATPase molecule was sufficient to inactivate the enzyme.

Adenosine Triphosphatases↗

Distribution of nociceptive neurons in the ventrobasal complex of macaque thalamus.

In urethane-chloralose anesthetized Japanese macaques, the distribution of nociceptive neurons within the thalamic ventrobasal (VB) complex was studied. Nociceptive specific (NS) and wide dynamic range (WDR) neurons were found in the periphery of the contralateral integument compartment of the VB complex. Thus, they formed a shell at the perimeter of this compartment with a somatotopic organization. The compartment consisted of large parts of nucleus ventralis posteromedialis (VPM) and nucleus ventralis posterolateralis, pars caudalis (VPLc). NS neurons were located more caudally than WDR neurons. In the NS zone of VPM, the forehead was represented caudally, and oral structures rostrally. In the ventral NS zone of VPM, there was a sequential representation of the tongue, gum and mandibular skin from the medial to the lateral edge. The hand was represented medially in the NS zone of VPLc, and its representation dominated in the rostral NS zone. There was a sequential representation of the cervical, thoracic, lumbar, sacral and caudal segments mediolaterally along the dorsal VPLc. In the medial half of ventral NS zone of VPLc, the upper body half was represented, and in the lateral half, the lower body half. The foot was represented at or near the medial edge of lateral half. In the rostral WDR zone, the trunk and peripheral face were represented.

Animals↗

Enhancement of retroviral gene transduction on a dish coated with a cocktail of two different polypeptides: one exhibiting binding activity toward target cells, and the other toward retroviral vectors.

CH-296, a recombinant fragment of human fibronectin (FN) composed of the cell-binding domain (C-domain), heparin-binding domain II (H-domain), and CS1 site, enhances the retrovirus-mediated gene transduction (GT) of hematopoietic stem cells. The RGD sequence in the C-domain is recognized by a variety of cell types through integrin VLA-5, and the LDV sequence in the CS1-site is recognized by integrin VLA-4. Retrovirus particles were also found to bind to the H-domain. Consequently, the CH-296 fragment can enhance GT through binding to both retrovirus particles and target cells that express integrins VLA-5 and/or VLA-4. In this study, we found that the GT efficiency can be maintained at levels comparable to that of CH-271, a FN fragment similar to CH-296 but lacking the CS1 site, when a cocktail of separated functional domains of CH-271 is used. When a dish was coated with a mixture of the C-domain and H-domain (molar ratio, 1:10), the GT efficiency of NIH3T3 cells reached the same level as that of the mother fragment, CH-271. The H-domain in the cocktail can be replaced with other virus-binding components, polylysine, FGF, and the insulin-binding domain of ColV, without the loss of GT efficiency. With other than FN fragments, a cocktail of erythropoietin and polylysine caused higher GT efficiency of Epo-receptor expressing TF-1 cells than in the case of each component alone.

Animals↗

Development of flat adenoma and superficial rectal cancer after pelvic radiation.

Little is known about the early development of rectal cancer in patients with radiation-induced colitis. We describe two patients with a history of radiation colitis who developed rectal cancer. The macroscopic appearance of these lesions suggested that they arose de novo, but the histologic findings were more consistent with progression along the adenoma-to-carcinoma sequence, a hybrid of de novo lesion and adenoma. Early rectal cancer associated with chronic inflammation should be considered if a nonpolypoid adenomatous lesion is detected in a patient who has received pelvic radiation.

Adenocarcinoma, Mucinous↗

A case of atypical chronic myeloid leukemia regarded as MDS with myeloproliferative features.

We report a case of atypical chronic myeloid leukemia (aCML) who showed marked neutrophilia without dysplastic features, basophilia or monocytosis. These findings diverged somewhat from the FAB criteria for aCML. The patient's erythroid cells and megakaryocytes were dysplastic. His marrow cells formed no spontaneous colonies, as shown by cell culture. The cells formed many small-sized neutrophil colonies with G-CSF stimulation. Interestingly, they formed mainly neutrophil colonies with GM-CSF stimulation. These findings were different from those of chronic myelomonocytic leukemia cells and chronic granulocytic leukemia cells. This aCML case showed the cytological features of myelodysplastic syndrome.

Humans↗

Synthesis of new peptides with prolactin-releasing activity by a combination of recombinant DNA technology and a cysteine-specific cyanylation reaction.

A newly isolated peptide from bovine hypothalamus with prolactin-releasing activity (prolactin-releasing peptide; PrRP) was synthesized by a combination of recombinant DNA technology and a cysteine-specific cyanylation reaction, together with rat and human homologs. The peptides were expressed in the form of fusion proteins with basic fibroblast growth factor mutein, which were purified by heparin-affinity chromatography. The fusion proteins were cleaved at the cysteine residues of the junction site by cyanylation, followed by treatment with ammonia for C-terminal amidation. Purification of the resulting crude peptides was performed using chromatography on a gel-filtration column, a cation-exchange column, and a reversed-phase column. As an example, about 90 mg of bovine PrRP (bPrRP) was obtained from 201 of culture bloth. The purified b PrRP showed full biological activities in binding to its receptor expressed on CHO cells and releasing arachidonic acid metabolite from the same cells, while the C-terminal acid form of bPrRP had little of these activities. These results indicate that the C-terminal amide structure is very important for expressing biological activity. The peptides obtained here might be very useful for studies on their biological significance and roles in vivo.

Amino Acid Sequence↗

Superovulatory responses in Japanese black beef cows following largest follicle aspiration or human chorionic gonadotrophin (hCG) treatment.

The effect of largest follicle aspiration or hCG administration before induction of superovulation on the ovarian response of Japanese Black beef cows was investigated using a crossover design in which induction of superovulation was attempted in every cow. The superovulatory response of cows whose largest follicle had been aspirated from the ovaries by ultrasound-guided follicular aspiration 1 day before induction of superovulation, did not differ from the response in non-treated control cows. In contrast, in cows given 5,000 IU of hCG 3 days before induction of superovulation, the proportions of fertilized ova and transferable embryos significantly decreased compared with the other groups.

Animals↗

[MIDCAB procedure by original stabilizer and usual devices].

UNLABELLED: The key of MIDCAB is how to stabilize the left anterior descending artery. We introduce our MIDCAB procedure by using our original stabilizer and usual devices in this paper. METHOD: Between March 1997 and August 1997, 13 patients underwent left internal thoracic artery to left anterior descending artery revascularization without extracorporeal circulation, and one case underwent right internal thoracic artery to right coronary artery. In all cases, we exposed the internal thoracic artery through a 8 cm anterior fifth space thoracotomy. One case of MIDCAB to right coronary artery was added an inferior hemisternotomy for anastomosis to the right coronary artery. The internal thoracic artery was harvested by using the Kent retractor. The anastomosis were done by using our original stabilizer and retract O tapes. RESULTS: In our method, there was no mortality and no postoperative cardiac event. There were 5 postoperative complications, one was reoperation due to bleeding from a branch of the internal thoracic artery, two cases were wound infection. Cerebral infarction, bleeding due to urinary bladder tumor and lumbar compression fracture was seen in one case each. CONCLUSION: MIDCAB procedure was performed safety and effectively by using our original stabilizer and usual devices.

Aged↗

Application of cryoprecipitate as a hematostatic glue.

BACKGROUND: The effectiveness of cryoprecipitate, harvested from a patient's own fresh frozen plasma, for use in cardiac surgery as a hematostatic glue was studied in 32 randomized elective adult cardiac surgery patients from January 1993 to July 1994. MATERIALS AND METHODS: Patients from the Toho Sakura Hospital were randomly allocated to two groups: Group 1 (n=11) received conventional fibrin glue presently available in our institution; while Group 2 (n=21) received autologous cryoprecipitate as a hematostatic glue. Surgical procedures broken down by group were as follows: Group 1: 4 CABG, 5 valvular surgeries and 2 other. Group 2: 11 CABG, 6 valvular surgery, 4 other. We preserved the patient's own blood and stored pure red cell and fresh frozen plasma (FFP). Cryoprecipitate was prepared from the FFP and preserved until required. RESULTS: Cryoprecipitate had a 5-fold increase in fibrinogen activity (1190+/-311 mg/dl vs 238+/-34 mg/dl p<0.001), a 10-fold increase in factor VIII activity (362+/-219% vs 34+/-11%, p=0.001), and 4.5-fold increase in factor XIII activity (538+/-213% vs 119+/-50%, p<0.001), compared to serum. The amount of bleeding postoperatively was slightly lower in the cryoprecipitate glue group compared to the conventional glue group, but this was not significantly different. CONCLUSIONS: We conclude that autologous samples of human cryoprecipitate prepared from a patient's own FFP had a strong hematostatic effect compared to conventional fibrin glue and was a very valuable hematostatic agent during cardiac surgery.

Adhesives↗

Bovine oocyte diameter in relation to developmental competence.

This study was conducted to determine the diameter of bovine oocytes that were able to attain their full developmental competence to blastocysts. Oocytes were recovered by aspiration of surface-visible follicles (1 to 7 mm in diameter) from slaughterhouse ovaries. Only healthy-looking cumulus-oocyte complexes were used for in vitro maturation, and they were divided into six groups based on diameter: < 110 microm, 110 to < 115 microm, 115 to < 120 microm, 120 to < 125 microm, 125 to < 130 microm and >/= 130 microm. Oocytes were processed through standard procedures for in vitro maturation, fertilization and culture. Following in vitro maturation or fertilization, some oocytes were stained to assess nuclear maturation and penetration rates. The numbers of embryos that cleaved at 42 h post insemination and developed to blastocysts and hatched blastocysts after 8 days of culture were recorded. The mean oocyte diameters were 114.0 +/- 4.8 microm. The oocytes displayed size-related ability to undergo meiotic maturation. The rates of nuclear maturation of oocytes in the greater than 115-microm size range were significantly higher than those of oocytes with diameters < 115 microm. In the < 120 microm diameter groups, the polyspermic fertilization rates of oocytes < 115 microm were significantly higher than those of oocytes 115 to < 120 microm in diameter. The rates of cleavage and development to blastocysts and hatched blastocysts rose as oocyte diameter increased. Among oocytes with a diameter >110 microm, oocytes < 120 microm were found to have significantly lower developmental competence than oocytes 120 to < 130 microm in diameter. These results suggest that bovine oocytes have acquired full meiotic competence at a diameter of 115 microm but not yet attained full developmental competence to blastocysts, and that oocytes have acquired full developmental competence at a diameter of 120 microm.

Journal Article↗

The locus coeruleus of the Japanese monkey (Macaca fuscata) does not express mu-opioid receptor-like immunoreactivity.

It is well known that locus coeruleus (LC) of the rat shows intense mu-opioid receptor-like immunoreactivity (MOR-LI). In the course of our study on the distribution of MOR-LI in the brain of the Japanese monkey (Macaca fuscata), however, no MOR-LI was found in the LC although the distribution pattern of MOR-LI in other regions of the lower brainstem of the monkey was essentially the same as that observed in the rat. It was also found that immunoreactivity for Met-enkephalin, the most potent endogenous ligand for MOR, was intense in the rat LC, but very weak, if any, in the monkey LC. MOR may not be expressed in the monkey LC.

Amino Acid Sequence↗

Cryopreservation of mature bovine oocytes following centrifugation treatment.

In vitro matured bovine oocytes were frozen slowly in 1.6 M 1,2-propanediol following centrifugation treatment for polarization of lipid droplets in the cytoplasm. After thawing, the survival of the oocytes was assessed morphologically and also by in vitro fertilization and culture. The polarization of cytoplasmic lipid droplets had a negative effect on the survival of frozen-thawed oocytes. Thus, this treatment did not improve the frequency of normal fertilization and development to blastocysts, compared with that of frozen control oocytes. However, the frequency of polyspermy and activation of lipid-polarized oocytes that survived after freezing-thawing and subsequent in vitro fertilization tended to be less than those of surviving control oocytes. In addition, the effect of centrifugation treatment was to produce a small but significant increase in the cleavage rate of oocytes that survived after freezing-thawing and the development rates to blastocysts of surviving lipid-polarized oocytes tended to increase, compared with those of surviving control oocytes. These results suggest that the freezing tolerance of the spindle and other organelles of in vitro matured bovine oocytes is associated with lipid droplets and may be improved by the polarization of cytoplasmic lipid droplets before cryopreservation.

Animals↗

Cultured retinal pericytes stimulate in vitro angiogenesis of endothelial cells through secretion of a fibroblast growth factor-like molecule.

Interaction between cultured endothelial cells (EC) and pericytes (PC) was studied in vitro to clarify the mechanism of diabetic proliferative retinopathy. Conditioned medium (CM) from retinal PC strongly increased the proliferation and moderately stimulated migration of retinal EC. Moreover, CM from PC caused stimulation of angiogenesis of retinal EC and umbilical cord vein EC in vitro at the same extent as basic fibroblast growth factor (bFGF). PC also stimulated angiogenesis by EC in mixed cultures. The angiogenic, proliferative and migration activities in CM from PC were inhibited by an antibody to bFGF. These data suggest that PC play an important role in angiogenesis through secretion of an FGF-like molecule.

Animals↗

Transcriptional regulation of the N-acetylglucosaminyltransferase V gene in human bile duct carcinoma cells (HuCC-T1) is mediated by Ets-1.

N-Acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to alpha-6-D-mannoside to produce the beta1-6 linked branching of N-glycan oligosaccharides, which controls the polylactosamine content. The expression of N-acetylglucosaminyltransferase V, which contains 17 exons and spans 155 kilobase pairs, is expressed in a tissue- and cell type-specific manner and is regulated at the level of transcription by multiple promoters (Saito, H., Gu, J., Nishikawa, A., Ihara, Y., Fujii, J., Kohgo, Y., and Taniguchi, N. (1995) Eur. J. Biochem. 233, 18-26). To elucidate the mechanism by which the GnT-V gene is expressed in a cell- and tissue-specific manner, cell-restricted expression was analyzed using the 5'-upstream regions of the human GnT-V gene spanning base pairs -2760 to +23 in a human bile duct carcinoma cell line, HuCC-T1. We characterized two cis-acting elements that are potentially important in HuCC-T1 cell-specific expression. The two elements each contain an Ets-1 binding site, 5'-GGA-3'. Specific binding of Ets-1 to the respective elements was demonstrated by competition analysis as well as by antibody supershift experiments. Cotransfection of an Ets-1 expression plasmid along with a GnT-V promoter-luciferase reporter plasmid revealed the participation of Ets-1 in the regulation of the GnT-V gene transcription. These data indicated that the transcriptional regulation of the GnT-V gene was mediated by transcription factor Ets-1.

Bile Duct Neoplasms↗