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Biomedical subjects

S Arai

Publications and source records attributed to S Arai.

At least 127 records · Page 7Linked to original sources

Prior muscular contraction enhances disposal of glucose analog in the liver and muscle.

To noninvasively investigate in vivo glucose disposal in muscle and liver after exercise, 19F magnetic resonance spectroscopy (19F-MRS) was applied using 3-fluoro-3-deoxy-D-glucose (3FDG) as a metabolic probe. After 30 minutes of muscle contraction of rabbit hindlimb by a 1-Hz electrical stimulation, 3FDG 250 mg/kg was injected intravenously and 19F-MRS was performed on the postcontracted hindlimb or the liver. Rabbits subjected to muscle contraction showed 1.5- and 1.7-fold higher peaks for 3FDG signal intensity in the liver and muscle than those not subjected to it. 3FDG was converted to 3-fluoro-3-deoxy-gluconic acid (3FGA) in the muscle and liver, and 3FDG oxidation was not affected by muscle contraction. During intraportal 3FDG infusion for 120 minutes at a dose of 2 mg x kg-1 x min-1 after termination of muscle contraction, the postcontracted rabbits showed a continuous increase in the signal intensity of 3FDG and a 2.1-fold higher total signal intensity of 3FDG than those not subjected to muscle contraction. In conclusion, 19F-MRS allows direct noninvasive observation of 3FDG disposal in rabbit muscle and liver. The increased intensity of 3FDG in the liver after muscle contraction suggests that exercise enhances disposal of the glucose analog in the liver, as well as in muscle, and these effects persist for at least 2 hours after exercise.

Animals↗

In vivo metabolism of the vitamin D analog, 22-oxacalcitriol: evidence for side-chain truncation and 17-hydroxylation.

After intravenous administration of the vitamin D3 analog, 22-oxacalcitriol (OCT), to normal rats plasma metabolites were investigated by HPLC, GC-MS and LC-MS. Five side-chain oxidation metabolites, 24R(OH)OCT, 24S(OH)OCT, (25R)-26(OH)OCT, (25S)-26(OH)OCT and 24oxoOCT, were identified by comparison with the corresponding synthetic compounds. These side-chain oxidation metabolites were similar to those of calcitriol [1alpha,25(OH)2 vitamin D3] described previously. Besides these five metabolites, two unique side-chain cleavage metabolites, 20S(OH)-hexanor-OCT and 17,20S(OH)2-hexanor-OCT, were identified as main metabolites in plasma by GC-MS and LC-MS using a specific chemical reaction. Our studies suggest that OCT is extensively metabolized and circulates in blood as a number of metabolites as well as unchanged OCT. This metabolism includes both unique pathways of C23-O22 cleavage and 17-hydroxylation, in addition to the side-chain oxidation metabolites similar to those of 1alpha,25-(OH)2D3.

Animals↗

In Situ Transmission Electron Microscope Observation of Melting of Aluminum Particles.

; The processes of melting and freezing of aluminum (Al) particles have been observed directly in a transmission electron microscope. The liquid phase nucleated preferentially at the surface of an Al particle, surrounding the crystalline solid at the onset of melting. The liquid phase then propagated inside the Al particle at the expense of the solid.

Journal Article↗

Apoptosis and suppression of tumor growth by artepillin C extracted from Brazilian propolis.

Artepillin C was extracted from Brazilian propolis. Artepillin C (3,5-diprenyl-4-hydroxycinnamic acid) has a molecular weight of 300.40 and possesses antibacterial activity. When artepillin C was applied to human and murine malignant tumor cells in vitro and in vivo, artepillin C exhibited a cytotoxic effect and the growth of tumor cells was clearly inhibited. The artepillin C was found to cause significant damage to solid tumor and leukemic cells by the MTT assay, DNA synthesis assay, and morphological observation in vitro. When xenografts of human tumor cells were transplanted into nude mice, the cytotoxic effects of artepillin C were most noticeable in carcinoma and malignant melanoma. Apoptosis, abortive mitosis, and massive necrosis combined were identified by histological observation after intratumor injection of 500 microg of artepillin C three times a week. In addition to suppression of tumor growth, there was an increase in the ratio of CD4/CD8 T cells, and in the total number of helper T cells. These findings indicate that artepillin C activates the immune system, and possesses direct antitumor activity.

Animals↗

Crystallization and preliminary X-ray diffraction studies of a rice cysteine proteinase inhibitor, Oryzacystatin-I.

Oryzacystatin-I from rice seeds was overexpressed in Escherichia coli, purified, and crystallized by the sitting-drop vapor diffusion method. Crystals obtained with 2-methyl-2,4-pentanediol as a precipitant exhibited space group I4122, with unit cell parameters of a = b = 100.0 A, c = 54.2 A, and diffracted up to 2.8 A resolution at 100 K. The crystals have one molecule per asymmetric unit.

Base Sequence↗

Functional expression and enzymatic properties of two Sitophilus zeamais cysteine proteinases showing different autolytic processing profiles in vitro.

To characterize in more detail the cathepsin L-like cysteine proteinases from Sitophilus zeamais (SCPs) cloned in our previous study [Matsumoto et al. (1997) J. Biochem. 121, 464-476], we established a system for their functional expression and purification using a glutathione S-transferase (GST) fusion gene vector from Escherichia coli. The proenzyme forms of two representative SCPs, proSCPc1 and proSCPg3, were expressed as GST-fusion proteins and purified on a glutathione Sepharose column. GST-proSCPc1 undergoes autoproteolytic cleavage into the mature form efficiently at acidic pH, and exhibits significant proteolytic activity toward various substrates including hemoglobin and Z-Phe-Arg-MCA. The enzymatic characteristics of the activated form of SCPc1 are similar to those of mammalian cathepsin L, but its pH optimum for the hydrolysis of hemoglobin is significantly lower. The other proSCP, GST-proSCPg3, which has a shorter COOH-terminal domain than SCPc1, undergoes almost no autolytic processing and shows only very slight proteolytic activity, although the other enzymatic characteristics of GST-proSCPg3 are similar to those of GST-proSCPc1.

Amino Acid Sequence↗

Characteristics of UVA-induced delayed tanning: seasonal variation.

We studied seasonal variation in UVA-induced delayed tanning and compared it to that in UVA + B-induced delayed tanning. The minimal tanning dose (MTD) and the minimal melanogenic dose (MMD) were determined one week after UVA exposure or UVA + B exposure, respectively. The MTD determined in the summer (June to August) was significantly higher than that in winter (December to February). In addition, while MTD in the winter correlated well with skin color, there was no correlation between MTD and skin color in summer. MTD tended to correlate with self-reported tanning history at the initiation of tanning in the winter, but they were not correlated with each other in the summer. The MMD measured in the summer was significantly lower than that in the winter. MMD correlated well with skin color in both seasons; the lighter the skin, the smaller the MMD. Also, a correlation was detected between MMD and self-reported tanning history. These results suggest that the susceptibilities of delayed tanning to UVA and UVA + B are affected by different factors.

Adult↗

Enteric lesions induced by different pseudorabies (Aujeszky's disease) virus strains inoculated into closed intestinal loops of pigs.

Three different strains of pseudorabies (Aujeszky's disease) virus were inoculated into ligated closed loops in the jejunum and ileum of five specific-pathogen-free pigs. Infected areas were compared with respect to distribution of histologic lesions and pseudorabies virus antigen. Two wild-type strains of pseudorabies virus produced enteric lesions consisting of necrosis of the subepithelial macrophages in the basilar crypt epithelium, necrosis of the lymphoid follicles in the Peyer's patches, degeneration of the epithelial cells in the crypt and villi, degeneration of the neuronal cells in the myenteric plexuses, and formation of intranuclear inclusion bodies on postinoculation days 2-4. Pseudorabies virus antigen was initially detected in subepithelial macrophages of the dome of Peyer's patches on postinoculation day 2 and subsequently extended to superficial epithelium and deeper into the lymphoid follicles and myenteric plexuses on postinoculation days 3-4. Many pseudorabies virus particles were also detected in the center of their necrotic foci. However, 1 mutant strain (ara-T-resistant) of pseudorabies virus did not produce enteric lesions. The results suggest that the primary target of infection by wild-type strains of pseudorabies virus might be the macrophages distributed in the subepithelial area of the dome Peyer's patches.

Animals↗

Antigenic and genetic diversities of Babesia ovata in persistently infected cattle.

Exploring the antigenic and genetic diversities of Babesia ovata, we obtained several field isolates from grazing cattle in the Okushiri island, Japan. Parasite isolation was greatly facilitated by using bovine red blood cell-substituted SCID mice (Bo-RBC-SCID mice), into which the blood samples of the cattle were inoculated. Isolates from different individuals within a herd of cattle were compared in immunoblot analysis with an anti-B. ovata serum and also in Southern blot analysis with a probe for the small subunit ribosomal RNA gene. In both analyses, the isolates exhibited banding patterns that were significantly different from each other. We were also able to obtain a series of parasite isolates from a single cow in different seasons of a nine months period, including winter when active vector ticks were not in the field environment. Different seasonal isolates showed different banding patterns in both immunoblot and Southern blot analyses. By contrast, these analyses detected little difference among the parasites that had been passed various times in Bo-RBC-SCID mice, where no specific immune responses should be generated. These results indicate that individual animals within a herd of cattle were infected with antigenically and genetically diversified populations of B. ovata, and that the parasites could persistently infect a single animal with dynamic change in their predominant subpopulations.

Animals↗

Higher sensitivity in induction of apoptosis in fibroblast cell lines derived from LEC strain rats to UV-irradiation.

The proportion of S-phase cells in a WKAH rat cell population decreased and that of G1-phase cells increased at 8 and 18 hr post-incubation following UV-irradiation, although no significant change was observed in the ratio of the proportion of S-phase to G1-phase cells in a LEC rat cell population. Thus, UV-radiation-induced delay in the progression from the G1 to S phase was observed in WKAH rat cells but was not apparent in LEC rat cells. The fraction of LEC rat cells containing a sub-G0 DNA content increased with post-incubation time after UV-irradiation, but not that of WKAH rat cells. The proportion of the sub-G0 fraction in LEC rat cells increased with increasing doses of UV-rays. Low molecular weight DNAs extracted from UV-irradiated LEC rat cells exhibited an intense DNA ladder pattern at 18 and 24 hr post-irradiation by electrophoretic analysis, but not those from UV-irradiated WKAH rat cells. These results showed a higher sensitivity of LEC rat cells in induction of apoptosis than that of WKAH rat cells to UV-irradiation, although there was no difference in the survival curves among the cell lines from LEC and WKAH rats after UV-irradiation.

Animals↗

Higher sensitivity in LEC rat cells to a topoisomerase II inhibitor, ellipticine.

A concentration of ellipticine, an inhibitor of topoisomerase II, required to reduce cell survival to 37% (D37) is used as an index to compare the cellular sensitivity. D37 values of LEC and WKAH rat cells were 1.2 and 2.2 microM, respectively. Thus, LEC rat cells were approximately 1.8-fold more sensitive than WKAH rat cells to ellipticine. There was no significant difference between the topoisomerase II activities in nuclear extracts of LEC and WKAH rat cells. These results suggested that the high sensitivity of LEC rat cells to ellipticine is not associated with the level of topoisomerase II activity.

Animals↗

Candida guilliermondii infection in SCID mice in association with the acceleration of the elimination of transfused human red blood cells.

An acceleration of the elimination of transfused human (Hu) red blood cells (RBC) was found in C.B-17 scid (SCID) mice that were kept in our facility. Yeast like organisms were isolated from their tap water just as a pure culture and the two isolates SW5 and SW6 were assigned to be Candida guilliermondii by analysing their generic small subunit ribosomal RNA sequences. To test whether the isolates are infectious in mice, we inoculated SCID and BALB/c mice orally with SW5 and observed them for 63 and 48 days, respectively. The yeasts were frequently recovered from oral swabs, feces and their tap water throughout the experiment. Although none of the mice developed clinical signs or histopathological changes, a positive sero-conversion was confirmed in 4 of 5 SW5-inoculated BALB/c mice. Moreover, a significant acceleration of Hu-RBC elimination in all of the SW5-infected SCID mice was demonstrated. We believe this to be the first report of an inapparent but significant outbreak of C. guilliermondii infection in mice.

Animals↗

[Outcomes of primary angioplasty in patients aged 85 years old and older who have acute myocardial infarction].

Outcomes of primary angioplasty in 6 elderly patients with acute myocardial infarction who were admitted to the hospital between July of 1994 and June of 1997 were reviewed retrospectively. Emergency coronary angiography was done in 7 of 16 patients (44%) who were at least 85 years old and primary angioplasty was done in 6 patients (38%). Dilatation was successful in all 6 patients. Congestive heart failure occurred in 4 patients and cardiogenic shock occurred in 1 patient, but no patient died during hospitalization. Blood transfusion and surgical resection were done in 1 patient because of a giant hematoma and pseudoaneurysm at the puncture site. Although the creatinine level increased after angioplasty in all 6 patients, dialysis therapy was not needed. All patients were alive and none had angina at follow-up (mean follow-up period = 16.5 months). Primary angioplasty was successful in patients at least 85 years old; both short-term and long-term outcomes were good. Primary angioplasty should be considered to be an effective treatment for acute myocardial infarction in people 85 years old and older.

Aged↗

[Reassessment of a combination of cerebrospinal fluid scintigraphy and nasal pledget counts in patients with suspected rhinorrhea].

A combination study of cerebrospinal fluid scintigraphy and nasal pledget counts was performed using 37 MBq of 111In-DTPA in 12 patients with suspected rhinorrhea. A pledget was inserted and dwelled in each nasal cavity for 6 hours, with the patient prone during at least 30 minutes. A total of 18 studies was implemented and nasal pledget counting method successfully diagnosed all of CSF rhinorrhea. Diagnosis was possible when pledget counts were greater than 1 kcpm. In patients with persistent, intermittent and occult/no nasal discharge, rhinorrhea was found in 100% (5/5), 60% (3/5), 25% (2/8), respectively. Two cases only exhibited positive scintigraphy. MRI or CT cisternography should be first performed in patients with persistent discharge, but in patients with intermittent/occult discharge pledget counting method might take priority of other diagnostic modalities. In conclusion, nasal pledget counting method is a simple and useful tool for detecting rhinorrhea.

Adolescent↗

Effect of FK506 on the activation state of hepatic macrophages in Propionibacterium acnes-treated rats.

Activated hepatic macrophages can provoke massive liver necrosis following endotoxin stimulation through microcirculatory disturbances due to sinusoidal fibrin deposition in rats pretreated with heat-killed Propionibacterium acnes. In these rats, FK506 (tachlorinus) administered 24 h before and at the time of endotoxin injection, significantly attenuated liver injury compared with the rats given no FK506. The effect of FK506 on hepatic macrophage activation and its action sites were studied in Propionibacterium acnes-treated rats. When rats received Propionibacterium acnes intravenously, hepatic-mRNA expression of interferon-gamma-inducing factor and interleukin-2 and splenic-mRNA expression of interferon-gamma were significantly increased compared with normal rats. Hepatic-mRNA expression of CD14, a receptor for lipopolysaccharide and its binding protein complex, was also increased preceding the expressions of the three cytokines in the liver and spleen. FK506 administration attenuated hepatic-mRNA expression of interleukin-2 and both superoxide anions as well as tumour necrosis factor-alpha production by hepatic macrophages, but did not change CD14-mRNA expression in Propionibacterium acnes-treated rats. It is suggested that a cytokine network through interferon-gamma-inducing factor, interferon-gamma and interleukin-2 may operate during activation of hepatic macrophages in rats treated with heat-killed Propionibacterium acnes, while CD14 expression on the cells may increase independently of this network. FK506 seems to attenuate such activation by suppressing hepatic interleukin-2 expression, without affecting CD14 expression on the cells.

Animals↗

Taste buds have a cyclic nucleotide-activated channel, CNGgust.

Cyclic nucleotide-gated (CNG) channels have been characterized as important factors involved in physiological processes including sensory reception for vision and olfaction. The possibility thus exists that a certain CNG channel functions in gustation as well. In the present study, we carried out reverse transcription-polymerase chain reaction and genomic DNA cloning and characterized a CNG channel (CNGgust) as a cyclic nucleotide-activated species expressed in rat tongue epithelial tissues where taste reception takes place. Several types of 5'-rapid amplification of cDNA ends clones of CNGgust cDNA were obtained with various 5'-terminal sequences. As the CNGgust gene was a single copy, the formation of such CNGgust variants should result from alternative splicing. The encoded protein was homologous to known vertebrate CNG channels with 50-80% similarities in amino acid sequence, and particularly homologous to bovine testis CNG channel and human cone CNG channel with 82% similarities. CNGgust was functional when expressed in human embryonic kidney cells, where it opened upon the addition of cGMP or cAMP. Immunohistochemical analysis using an antibody raised against a CNGgust peptide demonstrated the channel to be localized on the pore side of each taste bud in the circumvallate papillae, with no signal observed for degenerated taste buds after denervation of the glossopharyngeal nerve. All these results, together with the indication that cyclic nucleotides play a role gustatory signaling pathway(s), strongly suggest the involvement of CNGgust in taste signal transduction.

Amino Acid Sequence↗