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Biomedical subjects

A Ohta

Publications and source records attributed to A Ohta.

At least 109 records · Page 6Linked to original sources
PubMed1998

Reconstitution of immune systems in RAG2-/- mice by transfer with interleukin-12-induced splenic hematopoietic progenitor cells.

The administration of a high dose of IL-12 into the mice resulted in the induction of splenomegaly. From the flow cytometry analysis of cellularity in an enlarged spleen, it was demonstrated that Thyl.2-CD45RB-c-Kit + Sca-1 + Lin- hematopoietic progenitor cells markedly increased in IL-12-administered mouse spleen compared with untreated mouse spleen. The IL-12-induced hematopoietic progenitor cells showed a greatly enhanced colony-forming activity in CFU-granulocyte/macrophage (CFU-GM), blast-forming units-erythroid (BFU-E) and CFU-spleen (CFU-S) assay. Moreover, it was initially demonstrated that the transfer of IL-12-induced splenic hematopoietic progenitor cells into immunodeficient RAG2-/- mice caused a complete reconstitution of their immune functions including T- and B-cell-mediated immunity. Thus, the evidence that IL-12 has a capability of inducing hematopoietic progenitor cells possessing stem cell-like activity in vivo, indicated another important immunomodulating activity of IL-12 in immunotherapy.

Animals↗
PubMed1998

Enhanced efficacy of nilvadipine in hypertensives whose raised ambulatory blood pressure is sustained during sleep.

This study was designed to clarify the relationship between the antihypertensive effects of the calcium antagonist nilvadipine, and circadian changes in blood pressure. Based on measurements using an ambulatory blood pressure monitoring system (ABPM), 17 outpatients with untreated essential hypertension were divided into two groups: a sustained hypertensive group (with a fall in blood pressure during sleep < 10%, n = 7) and a waking time hypertensive group (with a fall in blood pressure during sleep > or = 10%, n = 10). During treatment with nilvadipine (8 mg/day, > or = 2 weeks), patients were reexamined by ABPM. The antihypertensive effect of nilvadipine was significantly and negatively correlated with the night time fall in blood pressure: this effect was significantly greater in the sustained hypertensive group than in the waking time hypertensive group. These data suggest that the long acting calcium antagonist nilvadipine has more potent antihypertensive effects in patients with sustained hypertension ("nondippers") than in those whose hypertension lessens during sleep ("dippers").

Aged↗
PubMed1998

Dietary fructooligosaccharides increase calcium absorption and levels of mucosal calbindin-D9k in the large intestine of gastrectomized rats.

BACKGROUND: In gastrectomized rats intestinal calcium absorption and bone calcium levels markedly decrease and the levels recover as a result of feeding fructooligosaccharides (FOS). In the present study we examined the effects of gastrectomy and dietary FOS on intestinal calbindin-D9k (CaBP) levels. METHODS: One group of rats was subjected to a sham operation and fed a control diet. Two other groups of rats were gastrectomized, and those in one group were fed the control diet, whereas those in the other group were fed a diet containing 10% FOS. Intestinal calcium, magnesium, and phosphorus absorption levels and intestinal CaBP levels were measured. RESULTS: Gastrectomy increased CaBP levels in the distal small intestine, cecum, and colorectum but markedly decreased calcium absorption. Dietary FOS increased CaBP levels in the cecum and colorectum in the case of gastrectomized rats and improved calcium absorption. CONCLUSIONS: The results suggest that dietary FOS not only improve intestinal calcium absorption but also serve to maintain local calcium homeostasis in the intestine by increasing mucosal CaBP levels in the large intestine of gastrectomized rats.

Animals↗
PubMed1998

Dietary fructooligosaccharides prevent osteopenia after gastrectomy in rats.

Postgastrectomy osteopenia is observed generally in humans. Fructooligosaccharides increase the absorption of calcium from the large intestine of healthy rats. Thus, we have examined whether they stimulate calcium absorption and prevent osteopenia in rats following total gastrectomy. Rats were subjected to either a sham surgical operation or Billoth II gastrectomy. Seven rats from each surgical treatment group were fed a control diet, and another seven rats of each treatment group were fed a diet containing fructooligosaccharides (75 g/kg diet) for 4 wk. For 5 d each week, feces were collected, and the calcium and phosphorus contents were measured for calculation of the absorption of these minerals. At the end of the experiment, the rats were killed and bones were collected. The net calcium absorption, calcium content and bone mineral density of the femur and tibia in gastrectomized rats fed the control diet were significantly less than those in sham-operated rats fed control diet. The net calcium absorption in rats fed the fructooligosaccharides diet was greater than that in rats fed control diet. Moreover, dietary fructooligosaccharides prevented the decrease in the calcium content and bone mineral density in gastrectomized rats. Dietary fructooligosaccharides enhanced calcium absorption and prevented the changes indicative of postgastrectomy osteopenia such as decreases in bone calcium content and bone mineral density in gastrectomized rats.

Absorption↗
PubMed1998

True calcium absorption in the intestine is enhanced by fructooligosaccharide feeding in rats.

Fructooligosaccharides (FOS) have been shown to stimulate apparent calcium absorption in the intestine. In this study, we examined the effect of FOS on true calcium absorption using the calcium balance in combination with the 45Ca kinetics method. Sixteen 45-d-old male Wistar rats were randomly divided into two groups, a control group (n = 8) and a FOS group (n = 8). The diet fed to the FOS group contained 5% FOS, at the expense of half of the sucrose in the control diet. After an adaptation period (3 d) and a free-access period (3 d) that were used to estimate the amount of food required for pair-feeding on the basis of calcium, all of the rats were pair-fed throughout the experiment from the age of 51 d. A constant amount of calcium was fed to the rats in each group (95 mg /d). At age 60 d, a 3-d metabolic study was started by the intravenous injection of 45Ca. Several variables were calculated on the bases of measurements of calcium intake, calcium in feces and serum, and 45Ca in feces, urine and serum. Both true and apparent calcium absorption in the intestine (Vad and Vna) and urinary calcium were significantly greater in rats that had been fed FOS. There were no differences between the groups in endogenous net calcium excretion into feces (Vf; Vad - Vna). The calcium balance was also enhanced by FOS. Calcium balance in the FOS group was significantly correlated with the absorbed calcium (r2 = 0.936, P < 0.01), as was that in the control group (r2 = 0.994, P < 0.01). These results suggest that the increased true calcium absorption and balance produced by FOS feeding might improve bone calcification.

Animals↗
PubMed1998

Dietary fructooligosaccharides prevent postgastrectomy anemia and osteopenia in rats.

Gastrectomized rats develop anemia and osteopenia, and ingestion of fructooligosaccharides leads to an increase in iron absorption and promotes recovery from anemia in iron-deficient rats. Laparotomized (sham-operated control) rats and totally gastrectomized (Billoth II) rats, in groups of 14 each, were fed a control diet without fructooligosaccharides or a diet containing fructooligosaccharides (75 g/kg of diet) for 6 wk. All rats received an intramuscular injection of vitamin B-12 every 2 wk. Tail blood was collected every week for determination of hematocrit and hemoglobin concentration. At the end of the experiment, the rats were killed and the femur and tibia were collected for measurement of bone mineral density (BMD). The hematocrit, hemoglobin concentration, hemoglobin regeneration efficiency, and BMD of both femurs and tibias were significantly lower in gastrectomized rats fed the control diet than in the other three groups. Dietary fructooligosaccharides prevented anemia and osteopenia in totally gastrectomized rats.

Anemia↗
PubMed1998

Dietary fructooligosaccharides change the concentration of calbindin-D9k differently in the mucosa of the small and large intestine of rats.

Previously, we confirmed that dietary fructooligosaccharides (FOS) increase calcium absorption in rats. In this study, we examined the influence of FOS feeding on the concentration of calbindin-D9k of several intestinal segments in rats. Rats in the control group were fed a diet without FOS. Rats in the other two groups were fed the diet containing FOS at either 50 or 100 g/kg for 10 d and subjected to a calcium absorption study. On the final day of feeding, the rats were killed and the entire intestine was removed. The intestinal mucosa was collected from four segments, i.e., the proximal and distal segments of the small intestine, the cecum and the colorectum, respectively. The apparent absorption of calcium increased dose dependently (r = 0.9256, P < 0.0001). Significant positive correlations between apparent calcium absorption and the relative amounts of calbindin in both large intestinal segments were observed (cecum, r = 0.8956, P = 0.0011; colorectum, r = 0.8828, P = 0.0016). Also, significant negative correlations between apparent calcium absorption and the relative amounts of calbindin-D9k in both small intestinal segments were observed (proximal, r = -0.7149, P = 0. 0304; distal, r = -0.8740, P = 0.0021). In conclusion, FOS feeding increases levels of calbindin-D9k in the large intestine, but decreases those in the small intestine. Moreover, these results suggest that part of the stimulatory effect of fructooligosaccharides relates to the transcellular route of calcium absorption in the large intestine of rats.

Absorption↗
PubMed1998

Isolation and characterization of EPD1, an essential gene for pseudohyphal growth of a dimorphic yeast, Candida maltosa.

Additional copies of the centromeric DNA (CEN) region induce pseudohyphal growth in a dimorphic yeast, Candida maltosa (T. Nakazawa, T. Motoyama, H. Horiuchi, A. Ohta, and M. Takagi, J. Bacteriol. 179:5030-5036, 1997). To understand the mechanism of this transition, we screened the gene library of C. maltosa for sequences which could suppress this morphological change. As a result, we isolated the 5' end of a new gene, EPD1 (for essential for pseudohyphal development), and then cloned the entire gene. The predicted amino acid sequence of Epdlp was highly homologous to those of Ggp1/Gas1/Cwh52p, a glycosylphosphatidylinositol-anchored protein of Saccharomyces cerevisiae, and Phr1p and Phr2p of Candida albicans. The expression of EPD1 was moderately regulated by environmental pH. A homozygous EPD1 null mutant showed some morphological defects and reduction in growth rate and reduced levels of both alkali-soluble and alkali-insoluble beta-glucans. Moreover, the mutant could not undergo the transition from yeast form to pseudohyphal form induced by additional copies of the CEN sequence at pH 4 or by n-hexadecane at pH 4 or pH 7, suggesting that EPD1 is not essential for yeast form growth but is essential for transition to the pseudohyphal form. Overexpression of the amino-terminal part of Epd1p under the control of the GAL promoter suppressed the pseudohyphal development induced by additional copies of the CEN sequence, whereas overexpression of the full-length EPD1 did not. This result and the initial isolation of the 5' end of EPD1 as a suppressor of the pseudohyphal growth induced by the CEN sequence suggest that the amino-terminal part of Epd1p may have a dominant-negative effect on the functions of Epd1p in the pseudohyphal growth induced by the CEN sequence.

Amino Acid Sequence↗
PubMed1998

A morphological and morphometrical study of the retina in aging SAM mice.

We investigated whether neuronal cell loss occurred as a part of normal aging of the retina in the Senescence-Accelerated Mouse, strains SAMP1 and SAMR1, and in the BALB/c mouse. All three strains showed age-related atrophy of the retina after histologically normal development. Morphometrical study revealed the following facts. The rate of loss of photoreceptor cells in the peripheral retina was greater than in the central retina in all three strains. In the central retina, the rate of loss of photoreceptor cells was greater in the SAMP1 and SAMR1 mice than in the BALB/c mice. In the peripheral retinal, the SAMR1 and SAMP1 strains had fewer cells than the BALB/c strain at all ages, but the rate of loss of these cells did not differ among the three strains. The rate of loss of ganglion cells did not differ between the peripheral and central retinas in the three strains. The SAMR1 and SAMP1 strains had fewer ganglion cells in the peripheral retina than the BALB/c strain at all ages. Because the rate of age-related loss of these cells in SAMP1 mice was not accelerated, and they were short-lived, SAMP1 mice did not show marked age-related loss. On the contrary, the SAMR1 mice showed a marked loss of photoreceptor cells and ganglion cells late in life because of their longer life span, and we propose that this strain is a suitable animal model for the study of mechanisms of age-related loss of neuronal cells in the retina.

Aging↗
PubMed1998

Inhibitory effect of 2,5-dimethylpyrazine on oxytocic agent-induced uterine hypercontraction of normal or pregnant female rats.

Intraperitoneal administration of 2,5-dimethylpyrazine (2,5-DMP) was found to inhibit oxytocin- and prostaglandin F2alpha-induced tetanic uterine contractions in normal or pregnant female rats. This suggests that 2,5-DMP may be used as a countercontraction agent or relaxant for preventing oxytocic agent-induced medical accident including uterine rupture or pressure death of the fetus due to uterine contractions.

Animals↗
PubMed1998

Cloning and characterization of a chitinase-encoding gene (chiA) from Aspergillus nidulans, disruption of which decreases germination frequency and hyphal growth.

We cloned a chitinase-encoding gene from Aspergillus nidulans by polymerase chain reaction using degenerated oligonucleotide primers designed from the conserved amino acid sequences among chitinases from yeasts and Rhizopus spp. The cloned gene, named chiA, encoded a polypeptide consisting of 660 amino acids. Disruption of chiA had no effect on hyphal or conidiophore morphology, but germination frequency and hyphal growth rate decreased substantially. Expression of chiA was investigated using Escherichia coli beta-galactosidase as a reporter enzyme. The beta-galactosidase activity was present during hyphal growth and increased twice as the conidiophores developed. In situ staining of beta-galactosidase activity found high expression in metulae, phialides, and conidia during conidiophore development, indicating that the expression of chiA is developmentally regulated. This is the first report to isolate a chitinase gene from A. nidulans and investigate its functions using the gene disruption technique and gene fusion methods in filamentous fungi.

Amino Acid Sequence↗
PubMed1998

Effect of magnetite on the hematocrit value in exsanguinated rats.

Magnetite prepared by an enzyme-dependent reaction gradually released iron ion into the acidic-to-neutral buffer solution. A preparatory experiment was performed to examine the efficiency of magnetite as an iron supplement. Feeding exsanguinated rats with being magnetite resulted in the hematocrit value being recovered without any serious adverse effect on the digestive organs.

Animals↗
PubMed1997

Manipulation of Th1/Th2 balance in vivo by adoptive transfer of antigen-specific Th1 or Th2 cells.

We have investigated the possibility that the Th1/Th2 balance in vivo may be modulated by adoptive transfer of Th1 or Th2 cells induced in vitro. Thl cells were induced from I-Ad-binding OVA323-339-specific T-cell receptor-transgenic (TCR-Tg) mouse spleen cells by culturing with OVA323-339 peptide and antigen presenting cells (APC) in the presence of IL-2, IL-12 and anti-IL-4 mAb. Th2 cells were induced from TCR-Tg mouse spleen cells by culturing with IL-2, IL-4 and anti-IL-12 mAb in addition to OVA323-339 plus APC. Immunomodulating activities of both Th1 and Th2 cells were determined by their effect on delayed type hypersensitivity (DTH) responses or cytokine production. No significant DTH responses (footpad swelling) were observed in untreated BALB/c mice following a single injection of OVA323-339-pulsed syngeneic spleen cells. However, adoptive transfer of Th1 cells into BALB/c mice induced strong dose dependent DTH responses in response to I-Ad-bound OVA323-339 but not unrelated peptide. In contrast, only slight DTH responses were detected in BALB/c mice transferred with Th2 cells. In parallel with the DTH responses, increased levels of serum IFN-gamma were demonstrated in mice adoptively transferred with Th1, while no significant increase was observed in Th2-transferred mice. In vitro analysis also demonstrated that both spleen cells and popliteal lymph node cells prepared from Th1-transferred mice showed Th1-type cytokine production, while cells obtained from Th2-transferred mice revealed Th2-dominant cytokine production. Such immune deviation induced by antigen-specific Th1 cells was demonstrated up to three months after cell transfer. Therefore, it may be possible to manipulate the Th1/Th2 balance in vivo by adoptive transfer of antigen-specific Th1 or Th2 cells.

Adoptive Transfer↗
PubMed1997

Accumulation of misfolded protein aggregates leads to the formation of russell body-like dilated endoplasmic reticulum in yeast.

RNAP-1, an aspartic proteinase from a filamentous fungus Rhizopus niveus, is secreted very efficiently in Saccharomyces cerevisiae. It is synthesized first as a precursor form with signal sequence and prosequence in its amino-terminus. Our previous study indicated that the prosequence of RNAP-I had important roles in its correct folding and secretion in yeast, and that a prosequence-deleted derivative of RNAP-I, delta pro, was not secreted but was retained and degraded in the yeast endoplasmic reticulum (ER). In the present study, we show that the accumulation of delta pro in the yeast ER caused elevated synthesis of ER resident chaperones, indicating that delta pro is recognized as an unfolded protein species in the ER. Our biochemical data demonstrated that delta pro formed aggregates which contained BiP, but not protein disulfide isomerase (PDI), in the ER. Immunoelectron microscopical analysis revealed that the delta pro aggregates were indeed visible as electron-dense regions in the ER and nuclear envelope. Such 'chaperone-associated misfolded protein bodies' were observed for the first time in yeast. Morphologies of the ER and nucleus were drastically altered by the accumulation of the delta pro aggregates. The ER lost its flat cisternal shape; the ER lumen extended aberrantly and the ER membrane irregularly proliferated. The misfolded delta pro proteins are probably sorted from the ordinary ER lumen to form the aggregates so that the ER function would not be grossly impaired, and the dilated ER may represent an ER subcompartment where the delta pro aggregates are degraded.

Amino Acid Sequence↗
PubMed1997

Structures of phenolic glycolipids from Mycobacterium kansasii.

From the lipid fraction of cells of Mycobacterium kansasii, four phenolic glycolipids K5, K6, K7 and K8, were isolated, in addition to three known phenolic glycolipids KI, KII and KIV. K5 was identified as a tetraglycosyl phenolic glycolipid whose sugar moiety was 2,6-dideoxy-4-O-methyl-L-alpha-arabinohexopyranosyl(1-->3)-4-O-pro pionyl-2-O-methyl-L-alpha-fucopyranosyl(1-->3)-2-O-methyl-L-alpha-rhamno pyranosyl-(1-->3)-2,4-di-O-methyl-L-alpha-rhamnopyranosyl(1-->) and phenolic glycolipid K6 as deacetyl-KI. Glycolipids K7 and K8 were triglycosyl phenolic glycolipids having the sugar moieties of 2-O-methyl-L-alpha-fucopyranosyl(1-->3)-2-O-methyl-L-alpha-rhamnopyranos yl(1-->3)-2,4-di-O-methy-L-alpha-rhamnopyranosyl(1-->) and 2-O-methyl-L-alpha-fucopyranosyl(1-->3)-2-O-methyl-L-alpha-rhamnopyranos yl(1-->3)-2-O-methyl-L-alpha-rhamnopyranosyl(1-->), respectively. Phenolic glycolipids K6 and K7 have been referred to as controlled degradation products of phenolic glycolipid KI previously. Also isolated was 5-mycolyl-beta-arabinofuranosyl(1-->2)-5-mycolyl-alpha-ar abinofurnosyl(1-->1')-glycerol, an analogue of glycolipid ai, originally isolated from Mycobacterium avium-Mycobacterium intracellulare complex, having mycolic acids of M. kansasii.

Antigens, Bacterial↗
PubMed1997

A novel fungal gene encoding chitin synthase with a myosin motor-like domain.

A csmA gene that encodes chitin synthase with a myosin motor-like domain was isolated from the filamentous fungus Aspergillus nidulans. Initially, we obtained the csmA as a homolog of the Aspergillus fumigatus chsE-partial fragment. A large open reading frame encoding a polypeptide of 1,852 a.a. was identified by determining the cDNA sequences. The chitin synthase conserved region was situated at the C-terminus and classified into class V as reported previously. On the other hand, the N-terminal region showed significant similarity to myosin motors and could not be classified into any types of myosins identified so far. Thus, it is suggested that this is the first report of unconventional myosin fused to a metabolic enzyme. The finding of this new type of chitin synthase gene suggests that localization of chitin synthesis may be guided by association with cytoskeletal structures.

Amino Acid Sequence↗
PubMed1997

The role of phorbol ester-sensitive protein kinase C isoforms in lymphokine-activated killer cell-mediated cytotoxicity: dissociation between perforin-dependent and Fas-dependent cytotoxicity.

Treatment of lymphokine-activated killer (LAK) cells with phorbol ester (PMA) caused the downmodulation of LAK activity concomitantly with the inhibition of serine esterase (SE) release, which has been shown as a marker for perforin-dependent cell-mediated cytotoxicity. The reduction of perforin-dependent LAK activity by PMA-treatment appeared to be due to the disappearance of PMA-sensitive protein kinase C (PKC) isoforms such as PKC alpha, gamma, epsilon, theta. In contrast, Fas-mediated LAK activity was refractory against PMA-induced downregulation. Treatment of LAK cells with PMA caused a disappearance of cytotoxicity against Fas L5178Y tumor cells, while cytotoxicity against Fas+ transfectants was not affected by PMA treatment. Moreover, Fas-mediated LAK activity of perforin-knockout mice was not inhibited by PMA treatment. These results clearly demonstrated that Fas-mediated cytotoxicity could be dissociated from perforin-mediated cytotoxicity by their different requirement of PMA-sensitive PKC isoforms.

Animals↗