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N Yoshida

Publications and source records attributed to N Yoshida.

At least 91 records · Page 5Linked to original sources

EMBRYONIC FLOWER2, a novel polycomb group protein homolog, mediates shoot development and flowering in Arabidopsis.

In higher plants, developmental phase changes are regulated by a complex gene network. Loss-of-function mutations in the EMBRYONIC FLOWER genes (EMF1 and EMF2) cause Arabidopsis to flower directly, bypassing vegetative shoot growth. This phenotype suggests that the EMF genes play a major role in repression of the reproductive program. Positional cloning of EMF2 revealed that it encodes a zinc finger protein similar to FERTILIZATION-INDEPENDENT SEED2 and VERNALIZATION2 of Arabidopsis. These genes are characterized as structural homologs of Suppressor of zeste 12 [Su(z)12], a novel Polycomb group gene currently identified in Drosophila. In situ hybridization studies have demonstrated that EMF2 RNA is found in developing embryos, in both the vegetative and the reproductive shoot meristems, and in lateral organ primordia. Transgenic suppression of EMF2 produced a spectrum of early-flowering phenotypes, including emf2 mutant-like phenotype. This result confirms the role of EMF2 in phase transitions by repressing reproductive development.

Arabidopsis↗

Development of a magnetic sensing device for tooth displacement under orthodontic forces.

We have developed a system for measuring tooth displacement from orthodontic force. Eight small magnetic sensors and a magnet are combined to measure three-dimensional displacement. Sensors, arranged cubically in the three planes of space, are placed in the mouth and fixed to the posterior teeth by a splint. A magnet is placed in the center of the eight sensors and attached to a front tooth that is subjected to orthodontic force. Sensors detect the magnet's movement as target tooth displacement. The system was designed to achieve displacement resolution of 1 microm. The mean percentage of measurement errors was determined to be less than 1% in a 600-cubic-microm volume from calibration. The system was tested clinically on human teeth. Although the oral environment, with high temperature and humidity, was not agreeable with the sensors, this system was stable and accurate enough for quantitative measurement of tooth displacement. The advantage of this system is the ability to detect tooth trajectories by decomposing displacement into translation and rotation and to determine the position of the center of rotation from these parameters.

Calibration↗

Modulation of ventricular repolarization and R-R interval is altered in patients with globally impaired cardiac 123I-MIBG uptake.

BACKGROUND: Cardiac (123)I-metaiodobenzylguanidine (MIBG) imaging is widely used to assess cardiac sympathetic neuronal function. However, physiologic significance of impaired cardiac MIBG uptake is not fully elucidated. The purpose of the present study was to determine influences of abnormal cardiac sympathetic neuronal function on heart rate variability (HRV) and ventricular repolarization process. METHODS: Twenty-nine patients with prior myocardial infarction were divided into two groups by a heart-to-mediastinum ratio (H/M) of MIBG scintigraphy. Ten patients with globally decreased MIBG uptake (group I: H/M < 1.5), 19 patients with partially decreased MIBG uptake (group II: H/M >or= 1.5), and 17 control subjects with normal MIBG uptake (group III) were studied. Holter recording and a standard 12-lead electrocardiography were used for evaluation of HRV, QT-RR relation, and QT dispersion. RESULTS: Low, high, and total frequency components decreased in groups I and II, as compared to that of group III. The reduction of these frequency domain measures was more severe in group I than in group II, but the differences did not reach statistical significance. Circadian variation of frequency domain measures disappeared in group I. The slope of QT-RR relation was significantly greater in group I than in groups II and III. QT dispersion was also greater in group I (64 +/- 25 msec) than in group II (43 +/- 19 msec) and group III (28 +/- 9 msec). CONCLUSION: These results suggest that patients with sympathetic neuronal dysfunction inferred from globally impaired cardiac MIBG uptake have an altered modulation of ventricular repolarization process as well as decreased HRV.

3-Iodobenzylguanidine↗

Targeted reduction in expression of Trypanosoma cruzi surface glycoprotein gp90 increases parasite infectivity.

A previous study had shown that the expression of gp90, a stage-specific surface glycoprotein of Trypanosoma cruzi metacyclic trypomastigotes, is inversely correlated with the parasite's ability to invade mammalian cells. By using antisense oligonucleotides complementary to a region of the gp90 gene implicated in host cell adhesion, we investigated whether the selective inhibition of gp90 synthesis affected the capacity of metacyclic forms to enter target cells. Parasites were incubated for 24 h with 20 microM PS1, a phosphorothioate oligonucleotide based on a sequence of the gp90 coding strand; PS2, the antisense counterpart of PS1; or PO2, the unmodified version of PS2 containing phosphodiester linkages, and the expression of surface molecules was analyzed by flow cytometry and immunoblotting using specific monoclonal antibodies. PS2 but not PS1 or PO2 inhibited the expression of gp90. Inhibition by PS2 was dose dependent. Northern blot analysis revealed that steady-state gp90 mRNA levels were diminished in PS2-treated parasites compared to untreated controls. Treatment with PS2 did not affect the expression of other metacyclic stage surface glycoproteins involved in parasite-host cell interaction, such as gp82 and the mucin-like gp35/50. Expression of gp90 was also inhibited by other phosphorothioate oligonucleotides targeted to the gp90 gene (PS4, PS5, PS6, and PS7) but not by PS3, with the same base composition as PS2 but a mismatched sequence. Parasites treated with PS2, PS4, or PS5 entered HeLa cells in significantly higher numbers than untreated controls, whereas the invasive capacity of PS1- and PS3-treated parasites was unchanged, confirming the inverse association between infectivity and gp90 expression.

Animals↗

Fast, but not slow, effects of olivocochlear activation are resistant to apamin.

Olivocochlear (OC) efferent suppression of auditory-nerve responses comprises a fast effect lasting tens of milliseconds and a slow effect building and decaying over tens of seconds. Both fast and slow effects are mediated by activation of the same alpha 9 nicotinic receptor. We have hypothesized that fast effects are generated at the OC synapse, but that slow effects reflect activation of calcium-activated potassium (K(Ca)) channels by calcium release from the subsurface cisternae on the basolateral wall of the hair cells. We measured in vivo effects of apamin, a blocker of small-conductance (SK) K(Ca) channels, and charybdotoxin, a blocker of large-conductance K(Ca) channels, perfused through scala tympani, on fast and slow effects evoked by electrical stimulation of the OC bundle in anesthetized guinea pigs. Apamin selectively and reversibly reduced slow-effect amplitude without altering fast effects or baseline amplitude of the auditory-nerve response, but only when perfused at concentrations of 100 microM. In contrast, the effects of charybdotoxin were noted at 30 nM, but were not specific, reducing both afferent and efferent responses. The very high concentrations of apamin needed to block efferent effects contrasts with the high sensitivity of isolated hair cells to apamin's block of acetylcholine's effects. The results suggest that in vivo fast OC effects are dominated by a conductance that is not apamin sensitive.

Action Potentials↗

Interleukin-4 is needed for the infection of monocytes by human cytomegalovirus.

OBJECTIVE: We studied the ability of human cytomegalovirus (HCMV) to infect peripheral blood mononuclear cells (PBMC) pretreated with or without Th2-cytokine interleukin-4 (IL-4) in vitro. METHODS: Adherent cells and nonadherent cells were obtained from PBMC. We inoculated these cells with HCMV at concentrations ranging from 0 to 10 ng/ml of IL-4. Immediate-early antigen-1 (IE-1) and glycoprotein H (gH) mRNAs were detected using the reverse-transcription polymerase chain reaction. RESULTS: IE-1 and gH mRNAs could be detected in monocytes pretreated with IL-4. In contrast, no IE-1 mRNA was detected in monocytes pretreated without IL-4. We tested whether higher infectious titers could result in the infection of monocytes whether or not they were pretreated with IL-4. However, no IE-1 mRNA was detected in the monocytes not pretreated with IL-4. To elucidate how HCMV-infected monocytes affect lung tissue, human embryonic lung fibroblasts MRC-5 were cocultured with HCMV-infected monocytes. The cytopathic effects of HCMV were observed microscopically and was confirmed by direct immunoperoxidase staining with a human monoclonal antibody against the HCMV IE-1. CONCLUSION: Our data strongly suggest that the ability of HCMV to infect monocytes may correlate with the presence of IL-4.

Cytomegalovirus↗

Role of P-selectin and intercellular adhesion molecule-1 in TNB-induced colitis in rats.

It has been proposed that neutrophil-endothelial cell interactions mediated by adhesion molecules are involved in the pathogenesis of inflammatory bowel disease. The objective of the present study was to determine the effects of monoclonal antibodies (MAbs) directed against endothelial adhesion molecules, P-selectin and intercellular adhesion molecule-1 (ICAM-1), in rats with colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNB). Colonic inflammation was induced by administering an enema of TNB dissolved in 50% ethanol (120 mg/ml) to male Wistar rats (at a total volume of 0.25 ml per rat) after a 48-hour fast. Anti-P-selectin MAb or anti-ICAM-1 MAb was injected via the tail vein at a dose of 1 mg/kg after the induction of colitis. Rats in the control group received nonbinding mouse immunoglobulin G1. The plasma level of soluble P-selectin showed an increase within 48 h after the TNB enema. Colonic inflammation was assessed at 1 week after TNB administration. The colonic damage score and the wet weight of the colon were significantly decreased by treatment with either MAb. The increase of myeloperoxidase (MPO) activity, an index of neutrophil accumulation, and the increase of thiobarbituric acid-reactive substances (TBA-RS), an index of lipid peroxidation, in the colonic mucosa were inhibited by both MAbs. These results suggest that neutrophil-endothelial cell interactions via P-selectin and ICAM-1 play an important role in the development of TNB-induced colitis in rats.

Animals↗

A549 cells can express interleukin-16 and stimulate eosinophil chemotaxis.

Alveolar epithelial cells produce many types of chemokines such as regulated on activation, normal T cells expressed and secreted (RANTES), eotaxin induced by interleukin (IL)-1 beta, or tumor necrosis factor (TNF)-alpha and may contribute to allergic disease by recruiting eosinophils. However, identification of the eosinophil chemotacic activity (ECA) release from A549 cells, an alveolar type II cell line, has not yet been completed. Recently, IL-16 was also reported to be a potent chemotactic stimulus for CD4(+) T lymphocytes and eosinophils in asthma and other pulmonary diseases. To test the possibility that alveolar epithelial cells produce IL-16, we analyzed RNA and culture supernatant from A549 cells by reverse transcription/ polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). The release of ECA from A549 cells was assessed using a blind-well chemotactic chamber. IL-16 release was increased in a concentration-dependent manner by stimulation with IL-1 beta or TNF-alpha. A549 cells also expressed IL-16 messenger RNA. The combination of IL-4 and IL-1 beta or TNF-alpha had an additive effect on IL-16 production. The release of ECA was induced by IL-1 beta or TNF-alpha in a dose-dependent manner. The combination of these cytokines had a greater effect than one alone. The blockade of eotaxin and IL-16 caused 70% inhibition of ECA, but anti-RANTES antibodies only caused 30% inhibition and anti-IL-8 antibodies failed to affect inhibition. These findings suggest a role for chemokines released by alveolar epithelial cells in the recruitment of eosinophils into the lung in pulmonary disorders such as asthma and interstitial lung diseases, and suggested that eotaxin and IL-16 are potent and effective eosinophil chemoattractants.

Base Sequence↗

Dietary phosphorus deprivation induces 25-hydroxyvitamin D(3) 1alpha-hydroxylase gene expression.

Dietary phosphorus deprivation causes hypophosphatemia and an increase in serum 1alpha,25-dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)] concentrations. To determine the molecular mechanisms of this regulation, the effects of dietary phosphorus deprivation and hypophysectomy on 25-hydroxyvitamin D(3) 1alpha-hydroxylase (1alpha-hydroxylase) protein and messenger RNA (mRNA) expression were examined in rats. A low phosphorus diet (LPD) for 4 days resulted in hypophosphatemia and an increase in serum 1,25-(OH)(2)D(3) levels. This increase was caused by the induction of 1alpha-hydroxylase protein and mRNA expression (4- and 10-fold increases, respectively). Administration of the LPD or normal phosphorus diet to hypophysectomized (HPX) rats resulted in hypophosphatemia and suppression of 1alpha-hydroxylase gene expression, indicating that hypophosphatemia itself is not sufficient to induce 1alpha-hydroxylase mRNA expression. Administration of GH to HPX rats fed LPD could partially restore 1alpha-hydroxylase mRNA expression, whereas supplementation with insulin-like growth factor I, T(3), estrogen, or corticosterone had no effect. We also examined Phex gene expression in the bone, because the clinical features of X-linked hypophosphatemia resemble those of HPX rats. Phex mRNA expression, however, was not altered in HPX rats. In conclusion, we demonstrated that the increase in serum 1,25-(OH)(2)D(3) levels caused by dietary phosphorus deprivation is due to the induction of 1alpha-hydroxylase mRNA expression, and this increase is mediated in part by a GH-dependent mechanism.

25-Hydroxyvitamin D3 1-alpha-Hydroxylase↗

Gene expression profiles in Ciona intestinalis tailbud embryos.

A set of 3423 expressed sequence tags derived from the Ciona intestinalis tailbud embryos was categorized into 1213 independent clusters. When compared with DNA Data Bank of Japan database, 502 clusters of them showed significant matches to reported proteins with distinct function, whereas 184 lacked sufficient information to be categorized (including reported proteins with undefined function) and 527 had no significant similarities to known proteins. Sequence similarity analyses of the 502 clusters in relation to the biosynthetic function, as well as the structure of the message population at this stage, demonstrated that 390 of them were associated with functions that many kinds of cells use, 85 with cell-cell communication and 27 with transcription factors and other gene regulatory proteins. All of the 1213 clusters were subjected to whole-mount in situ hybridization to analyze the gene expression profiles at this stage. A total of 387 clusters showed expression specific to a certain tissue or organ; 149 showed epidermis-specific expression; 34 were specific to the nervous system; 29 to endoderm; 112 to mesenchyme; 32 to notochord; and 31 to muscle. Many genes were also specifically expressed in multiple tissues. The study also highlighted characteristic gene expression profiles dependent on the tissues. In addition, several genes showed intriguing expression patterns that have not been reported previously; for example, four genes were expressed specifically in the nerve cord cells and one gene was expressed only in the posterior part of muscle cells. This study provides molecular markers for each of the tissues and/or organs that constitutes the Ciona tailbud embryo. The sequence information will also be used for further genome scientific approach to explore molecular mechanisms involved in the formation of one of the most primitive chordate body plans.

Animals↗

Effects of ovariectomy on intramuscular energy metabolism in young rats: how does sports-related-amenorrhea affect muscles of young female athletes?

The purpose of this study was to evaluate the effects of ovariectomy on intramuscular energy metabolism in young rats. Twenty-four Sprague-Dawley rats (7 weeks old) were used. Twelve of them underwent ovariectomy (OVX), and the others were sham-operated on. Seven OVX rats were examined 1-week after surgery (OVX-1 group), and the other five, 4 weeks after surgery (OVX-4 group). The gastrocnemius-plantaris-soleus (GPS) muscles group was subjected to the following measurements, and the data were compared with those of the sham group (Sham-1: n = 7, or Sham-4 group: n = 5). From the 31P-MR spectra of the GPS muscles group at rest and during electric stimulation, the muscular oxidative capacity was measured. Maximum tension and wet weight of the whole GPS muscles group were also measured. Body weight in the OVX-4 group was significantly (p < 0.01) larger than that in the Sham-4 group. The weights of the whole GPS muscles group in the Sham-1, Sham-4, OVX-1 and OVX-4 groups were 1.17, 1.51, 1.25 and 1.71 (g), respectively. The muscle weight in the OVX group tended to be greater than that in the Sham group (p < 0.10). The maximum tension and oxidative capacity did not differ significantly among the groups. These data indicated that in young rats, ovariectomy induced an increase in body and muscle weight, but did not affect the maximum tension nor oxidative capacity.

Amenorrhea↗

Evaluation of disuse atrophy of rat skeletal muscle based on muscle energy metabolism assessed by 31P-MRS.

The purpose of this study was to evaluate disuse atrophy of skeletal muscle using a hind-limb suspension model, with special reference to energy metabolism. Twenty-four Sprague-Dawley rats were divided into four groups: control group (C), hind-limb suspended for 3 days (HS-3), for 7 days (HS-7) and for 14 days (HS-14). The gastrocnemius-plantaris-soleus (GPS) muscles in each group were subjected to the following measurements. After a 2-min rest, contraction of the GPS muscles was induced by electrical stimulation of the sciatic nerve at 0.25 Hz for 10 min, then the frequency was increased to 0.5 and 1.0 Hz every 10 min. During the stimulation, twitch forces were recorded by a strain gauge, and 31P-MRS was performed simultaneously. Maximum tension was measured at the muscle contraction induced at 0.25 Hz; the wet weight of the whole and each muscle in the GPS muscles was also measured. From the 31P-MR spectra during muscle contraction, the oxidative capacity was calculated and compared among the groups. The weights of the whole GPS muscles in C, HS-3, HS-7 and HS-14, were 2.66 +/- 0.09, 2.39 +/- 0.21, 2.34 +/- 0.21 and 2.18 +/- 0.14 (g) respectively. Thus, the muscle mass significantly decreased with time (p < 0.05). Among the GPS muscles, the decrease in weight of the soleus muscle was especially remarkable; in the HS-14 group its weight decreased to 60% of that in the C group. We evaluated maximum tension and oxidative capacity as the muscle function. The maximum tensions in C, HS-3, HS-7 and HS-14 were 519 +/- 43, 446 +/- 66, 450 +/- 23 and 465 +/- 29 (g), respectively. This was significantly greater in the C group than in any other groups, however there were no significant differences among the three HS groups. The oxidative capacity during muscle contraction in the C group was higher than in any HS group and it did not further decrease even if the suspension of the limbs was prolonged beyond 3 days. The present study showed that in disuse atrophy, muscle mass and muscle function did not change simultaneously. Thus, it is necessary to develop countermeasures to prevent muscle atrophy and muscle function deterioration independently.

Animals↗

Role of gamma/delta T-cells in the peripheral blood of patients with pulmonary tuberculosis.

Although gamma/delta T-cells are known to contain the highest frequency of mycobacteria-reactive cells in humans, and recent studies have suggested that they play an important role in the initial immune response to Mycobacterium tuberculosis (Mtb), very few studies have attempted to analyze these cells in patients with active pulmonary tuberculosis (TB). The aim of the present study was therefore to evaluate the gamma/delta T-cell populations present in the peripheral blood and the IFN-gamma production of gamma/delta T-cells stimulated by PMA before and after anti-TB chemotherapy in patients in the initial treatment stage for primary active pulmonary TB. Cell populations were measured by three-color flow cytometry of peripheral blood mononuclear cells. We compared the population of gamma/delta T-cells and the production of IFN-gamma between normal healthy controls and TB patients. Absolute numbers of gamma/delta T-cells remained constant in the peripheral blood of TB patients. However, production of IFN-gamma in gamma/delta T-cells was dramatically suppressed prior to anti-TB chemotherapy in comparison with healthy control subjects, and further reduced following anti-TB chemotherapy. We also examined the influence of isoniazid (INH) in anti-TB chemotherapy. INH suppressed IFN-gamma production of gamma/delta and alpha/beta T-cells. The findings demonstrated a strong correlation between the production of IFN-gamma in gamma/delta T-cells and manifestation of primary active pulmonary TB, which was consistent with the hypothesized role for gamma/delta T-cells in the protective immune response to Mtb infection.

Adult↗

In vitro transmission of HCMV between fibroblasts and peripheral blood leukocytes in the presence of IL-4.

We demonstrated transmission of human cytomegalovirus (HCMV) from the human lung fibroblast MRC-5 to peripheral blood leukocytes (PBLs). mRNA of the HCMV immediately-early (IE) antigen was detected in PBLs cultured with IL-2 or IL-2 + IL-4 that made direct contact with HCMV-infected MRC-5, whereas it was not detected in PBLs prevented from making cell-to-cell contact. However, mRNA of HCMV IE was not detected in PBLs cultured with IL-2 and IFN-gamma that made direct contact with HCMV-infected MRC-5. Transmission of the pp65 antigen was increased in culture medium containing IL-4. At a higher viral infection titer, cell-free HCMV infected adherent PBLs cells. The subset, which did not adhere, did not infect cell-free viruses even at a very high multiplicity of infection. Moreover, the adhered subset of PBLs infected with HCMV was able to transmit HCMV to non-infected fibroblasts. Our results suggest that cell-to-cell contact (when PBLs make direct contact with HCMV-infected cells) is important in the mechanism of HCMV transmission and that the adherent cells of PBLs are one of the most important vehicles for HCMV infection. Moreover, we suggest that type 2 cytokines such as IL-4 enhance the transmission of HCMV to PBLs.

Antigens, Viral↗

[Progress in study for mechanism of gastric mucosal injury].

Inflammatory reactions mediated by leukocytes, cytokines, adhesion molecules, active oxygen species, nitric oxide(NO) or prostaglandins have been implicated in the pathogenesis of gastric mucosal injury induced by Helicobacter pylori(H. pylori) or nonsteroidal anti-inflammatory drugs(NSAID). The molecular mechanism of gastric mucosal injury has been investigated in view of cell proliferation, apoptosis or necrosis. Eradication therapy of H. pylori, Cox2 inhibitors or NO-releasing NSAID may become new strategy for protection against gastric mucosal injury.

Animals↗

[A case report of advanced hepatocellular carcinoma successfully treated with surgical multi-ablation therapy based on radio-frequency thermal ablation].

We report a patient with advanced hepatocellular carcinoma (HCC) who was successfully treated with multi-ablation therapy by the surgical method of laparotomy and partial thoracotomy. A 67-year-old man, who had undergone transcatheter arterial embolization (TAE) three times for advanced HCC with hepatitis C, presented at our hospital in May 2001. Although TAE had been performed 3 times starting 2 years earlier, interventional therapy for recurrent tumors is difficult because of stenosis and obstruction of regional hepatic arteries. Computed tomography of the liver revealed five tumors in both lobes and the tumor in segment VIII presented difficulty for percutaneous ablation treatment due to its closeness to the trunk of the hepatic vein. Multi-ablation therapy consisting of radio-frequency ablation, microwave ablation and ethanol injection was selected for this patient. By laparotomy and partial thoracotomy, we performed multi-ablation therapy safely and accurately for each tumor. After this treatment, 5 tumors in both lobes showed no viability on dynamic CT and the patency of the right hepatic vein was preserved. Six months after the treatment, one remnant mass appeared in segment V. An additional two sessions of percutaneous RFA were performed for this tumor. At the end of these treatments, enhanced CT revealed no viability of the tumors, and serum alpha-fetoprotein and PIVKA-II level dropped to the normal range. This case suggests that multiple ablation therapy with a surgical method is feasible and useful in cases of advanced hepatocellular carcinoma.

Aged↗