Search PubMed⌕ Search

Biomedical subjects

T Iwanaga

Publications and source records attributed to T Iwanaga.

At least 55 records · Page 3Linked to original sources

MTABC3, a novel mitochondrial ATP-binding cassette protein involved in iron homeostasis.

Atm1p, a mitochondrial half-type ATP-binding cassette (ABC) protein in Saccharomyces cerevisiae, transports a precursor of the iron-sulfur (Fe/S) cluster from mitochondria to the cytosol. We have identified a novel half-type human ABC protein, designating it MTABC3 (mammalian mitochondrial ABC protein 3). MTABC3 mRNA is ubiquitously expressed in all of the rat and human tissues examined. MTABC3 protein is shown to be present in the mitochondria, as assessed by immunoblot analysis and confocal microscopic analysis of subcellular fractions of Chinese hamster ovary cells stably expressing MTABC3. Accumulation of iron in the mitochondria, mitochondrial DNA damage, and respiratory dysfunction in the yeast ATM1 mutant strain (atm1-1 mutant cells) were almost fully reversed by expressing MTABC3 in these mutant cells. These results indicate that MTABC3 is a novel ortholog of the yeast and suggest an important role in mitochondrial function. Interestingly, the human MTABC3 gene has been mapped to chromosome 2q36, a region within the candidate locus for lethal neonatal metabolic syndrome, a disorder of the mitochondrial function associated with iron metabolism, indicating that MTABC3 is a candidate gene for this disorder.

ATP-Binding Cassette Transporters↗

Effect of germinated barley foodstuff administration on mineral utilization in rodents.

Germinated barley foodstuff (GBF), which is derived from the aleurone and scutellum fractions of germinated barley, is rich in water-insoluble dietary fiber, which reportedly modulates mineral absorption in vivo. This study was conducted to examine whether GBF affects mineral absorption. We performed three experiments. In experiment 1, female ICR mice (4 weeks old) were fed a diet supplemented with 10% GBF or cellulose (CE; as a control) for 24 weeks. Body weight and food intake were recorded twice weekly. On the last experimental day, serum biochemical parameters, including electrolyte parameters, were determined, and major digestive organs were examined histopathologically. In experiment 2, male Sprague-Dawley rats (5 weeks old) were fed the CE and GBF diets, with the mineral content of the CE diet adjusted to the same level as that in the GBF diet, for 14 days, and the intake and fecal excretion of minerals (calcium, Ca; magnesium, Mg: Iron, Fe; Phosphate, P) were determined for the last 3 days. In experiment 3, the binding of Ca and Mg to GBF or cellulose and the release of Ca and Mg from GBF or cellulose was measured in phosphate buffer (pH 6.8). Long-term administration of GBF did not affect the growth rate of mice. GBF did not inhibit the absorption of Ca and Mg, and rather promoted the absorption of Ca and Mg by the gastrointestinal tract. The absorption of Fe and P was not attenuated by GBF administration, and GBF contained only a low level of phytic acid. GBF did not inhibit mineral absorption, differing from the effect of other dietary fibers, and is expected to be useful as a dietary source of Ca and Mg.

Absorption↗

Inhibitory effect of TMK-688 on late asthmatic responses as well as T-cell and eosinophilic infiltration in guinea pigs with asthmatic reactions.

The effects of an oral anti-allergic agent, TMK-688, which inhibits 5-lipoxygenase, at doses of 3.2 and 10 mg/kg were studied in guinea pigs with dual-phase asthmatic response. We previously observed that pretreatment with TMK-688 inhibited the late asthmatic response (LAR) induced by ovalbumin inhalation exposure. The present study focused on the effect of TMK-688 on infiltration by T-cells and eosinophils. TMK-688 inhibited both T-cell and eosinophilic infiltration. These findings suggest that TMK-688 is effective in inhibiting infiltration of T-cells and eosinophilic chemotaxis, and thereby suppresses LAR.

Animals↗

cAMP-GEFII is a direct target of cAMP in regulated exocytosis.

Although cAMP is well known to regulate exocytosis in many secretory cells, its direct target in the exocytotic machinery is not known. Here we show that cAMP-GEFII, a cAMP sensor, binds to Rim (Rab3-interacting molecule, Rab3 being a small G protein) and to a new isoform, Rim2, both of which are putative regulators of fusion of vesicles to the plasma membrane. We also show that cAMP-GEFII, through its interaction with Rim2, mediates cAMP-induced, Ca2+-dependent secretion that is not blocked by an inhibitor of cAMP-dependent protein kinase (PKA). Accordingly, cAMP-GEFII is a direct target of cAMP in regulated exocytosis and is responsible for cAMP-dependent, PKA-independent exocytosis.

Animals↗

Atypical hyperplasia of choledocho-pancreatic duct epithelium in an Otsuka Long Evans Tokushima Fatty strain of rats.

Epithelial papillary hyperplasia of choledocho-pancreatic duct, associated with cellular atypism, was observed in Otsuka Long Evans Tokushima Fatty (OLETF) rats, a strain originally established as an animal model for non-insulin-dependent diabetes mellitus (NIDDM). To investigate the potential feasibility of OLETF rats as an animal model for pancreatic ductal carcinoma, we examined the pathological characteristics of ductal lesions in OLETF rats aged from 5 to 50 weeks. Hyperplastic lesions in OLETF rats became apparent after 10 weeks of age and increased in severity and frequency of atypical changes in hyperplastic epithelium appearing after 20 weeks. We compared ductal lesions from OLETF rats with those from age-matched Long Evans Tokushima Otsuka (LETO) rats, which share a similar genetic background with OLETF rats but do not develop NIDDM. While LETO rats also display a tendency toward ductal hyperplasia, lesions from OLETF rats were more numerous and larger in size than those from age-matched LETO rats. In addition, lesions from OLETF rats contained a significantly higher number of proliferating cell nuclear antigen-positive cells than those from LETO rats. Finally, lesions in OLETF rats were accompanied by inflammation, and the observed morphological alteration of lesions correlated well with the grade of inflammation.

Aging↗

Gq protein alpha subunits Galphaq and Galpha11 are localized at postsynaptic extra-junctional membrane of cerebellar Purkinje cells and hippocampal pyramidal cells.

Following cell surface receptor activation, the alpha subunit of the Gq subclass of GTP-binding proteins activates the phosphoinositide signalling pathway. Here we examined the expression and localization of Gq protein alpha subunits in the adult mouse brain by in situ hybridization and immunohistochemistry. Of the four members of the Gq protein alpha subunits, Galphaq and Galpha11 were transcribed predominantly in the brain. The highest transcriptional level of Galphaq was observed in cerebellar Purkinje cells (PCs) and hippocampal pyramidal cells, while that of Galpha11 was noted in hippocampal pyramidal cells. Antibody against the C-terminal peptide common to Galphaq and Galpha11 strongly labelled the cerebellar molecular layer and hippocampal neuropil layers. In these regions, immunogold preferentially labelled the cytoplasmic face of postsynaptic cell membrane of PCs and pyramidal cells. Immunoparticles were distributed along the extra-junctional cell membrane of spines, dendrites and somata, but were almost excluded from the junctional membrane. By double immunofluorescence, Galphaq/Galpha11 was extensively colocalized with metabotropic glutamate receptor mGluR1alpha in dendritic spines of PCs and with mGluR5 in those of hippocampal pyramidal cells. Together with concentrated localization of mGluR1alpha and mGluR5 in a peri-junctional annulus on PC and pyramidal cell synapses (Baude et al. 1993, Neuron, 11, 771-787; Luján et al. 1996, Eur. J. Neurosci., 8, 1488-1500), the present molecular-anatomical findings suggest that peri-junctional stimulation of the group I metabotropic glutamate receptors is mediated by Galphaq and/or Galpha11, leading to the activation of the intracellular effector, phospholipase Cbeta.

Amino Acid Sequence↗

VIP- and PACAP-induced salivary chromogranin A secretion in the isolated perfused submandibular gland of rats.

In the study reported in this paper, sensitive ELISA for rat CgA was developed using synthetic rat CgA(359-389) as antigen, N alpha-biotinylated glycylglycyl rat CgA(359-389), and antirat CgA(359-389) serum for the measurement of CgA-LI in rat saliva. CgA-LI in rat submandibular tissues and saliva was characterized by both immunohistochemical and immunochemical methods. Using isolated perfused rat submandibular gland. VIP at 0.1-1.0 nM in the presence of 0.1 microM ACh was found to cause CgA-LI secretion, whereas neither PACAP-27 nor PACAP-38 showed any effect on CgA secretion.

Amino Acid Sequence↗

Mode of action of ANG II on ion transport in guinea pig distal colon.

The effect of ANG II on mucosal ion transport and localization of ANG type 1 receptor (AT(1)R) in the guinea pig distal colon was investigated. Submucosal/mucosal segments were mounted in Ussing flux chambers, and short-circuit current (I(sc)) was measured as an index of ion transport. Serosal addition of ANG II produced a concentration-dependent (10(-9)-10(-5) M) increase in I(sc). The maximal response was observed at 10(-6) M; the increase in I(sc) was 164.4 +/- 11.8 microA/cm(2). The ANG II (10(-6) M)-evoked response was mainly due to Cl(-) secretion. Tetrodotoxin, atropine, the neurokinin type 1 receptor antagonist FK-888, and piroxicam significantly reduced the ANG II (10(-6) M)-evoked response to 28, 45, 58, and 16% of control, respectively. Pretreatment with prostaglandin E(2) (10(-5) M) resulted in a threefold increase in the ANG II-evoked response. The AT(1)R antagonist FR-130739 completely blocked ANG II (10(-6) M)-evoked responses, whereas the ANG type 2 receptor antagonist PD-123319 had no effect. Localization of AT(1)R was determined by immunohistochemistry. In the immunohistochemical study, AT(1)R-immunopositive cells were distributed clearly in enteric nerves and moderately in surface epithelial cells. These results suggest that ANG II-evoked electrogenic Cl(-) secretion may involve submucosal cholinergic and tachykinergic neurons and prostanoid synthesis pathways through AT(1)R on the submucosal plexus and surface epithelial cells in guinea pig distal colon.

Angiotensin II↗

Once-daily theophylline reduces serum eosinophil cationic protein and eosinophil levels in induced sputum of asthmatics.

BACKGROUND: Because eosinophilic airway inflammation is a characteristic feature of bronchial asthma, the treatment of airway inflammation is important in the management of asthma. Theophylline has been reported to reduce airway inflammation, in addition to its well-known bronchodilating effect. OBJECTIVE: In order to evaluate the effects of theophylline on airway inflammation, we investigated 48 subjects with mild and moderate asthma. METHODS: The patients were randomly divided into two groups, with or without theophylline treatment (control n = 24; theophylline, n = 24). We examined the level of serum eosinophil cationic protein (ECP), induced sputum samples, and peak expiratory flow (PEF) and obtained spirograms before and after 4 weeks of treatment with once-daily theophylline (200-600 mg/day) of subjects with mild or moderate asthma. RESULTS: Theophylline significantly increased morning and evening PEF and significantly decreased the diurnal variation of PEF. After treatment with theophylline, both serum ECP and the percentage of eosinophils in induced sputum were significantly decreased. In contrast, peripheral blood eosinophil count was unchanged after treatment with theophylline. CONCLUSIONS: These findings suggest that theophylline reduces airway inflammation and the severity of asthma, presumably via suppression of both eosinophil activity and subsequent eosinophil infiltration of the airways.

Adolescent↗

Distribution of sarcophytol A in soft coral of the Sarcophyton genus.

The distribution of sarcophytol-A in the Sarcophyton genus was investigated in seven samples belonging to S. glaucum (3 samples), S. infundibulifurme (2 samples), S. crassocaule (1 sample) and S. trocheliophorum (1 sample) that were collected on Ishigaki Island in Okinawa Prefecture. Sarcophytol-A was present in one sample each of S. glaucum and S. infundibulifurme. This study indicates that the composition of cembranoids in the Sarcophyton genus is not related with the respective species, but with the individual samples collected.

Animals↗

Molecular cloning of equine chromogranin A and its expression in endocrine and exocrine tissues.

Chromogranin A (CGA) is a member of a family of highly acidic proteins co-stored and co-released with catecholamines in the adrenal medullary cells as well as in other neurons and paraneurons. The nucleotide sequence encoding equine CGA was determined using RT-PCR and rapid amplification of complementary DNA (cDNA) ends (RACE) techniques. A total 1,828 bp of the nucleotide sequence reveals that equine CGA is a 448-residue protein preceded by an 18-residue signal peptide. Comparison of the amino acid sequence of equine CGA with those of human, porcine, bovine, mouse, rat and frog CGA showed high conservation at the NH2-terminal 1-77 amino acids regions (94.8%, 93.5%, 92.2%, 81.8%, 83.1% and 66.2%, respectively) and COOH-terminal 314-430 amino acids regions (90.6%, 81.4%, 90.6%, 80.5%, 83.3% and 39.0%, respectively), as well as a potential dibasic cleavage site, whereas the middle portion showed marked sequence variation (52.5%, 49.1%, 38.9%, 26.6%, 27.9% and 6.2%, respectively). Northern blot analysis and RT-PCR elucidated the tissue distribution of equine CGA mRNA. Its expression was confirmed not only in the adrenal medullary cells but also in other organs (cerebrum, cerebellum, pituitary gland, spinal cord, liver, thyroid gland, striated muscle, lung, spleen, kidney, parotid gland and sublingual gland). Further, in adrenal chromaffin cells and pituitary cells of the anterior-intermediate lobe, the expression was confirmed by in situ hybridization with anti-sense CGA cRNA probe.

Amino Acid Sequence↗

Differential distribution of oestrogen receptor-alpha and -beta mRNAs in the female reproductive organ of rats as revealed by in situ hybridization.

The cellular distribution of two oestrogen receptor (ER) subtypes, ERalpha and ERbeta mRNAs, was studied in the entire female reproductive organ of the rat using in situ hybridization. Expression of ERalpha and ERbeta mRNAs was predominant in the reproductive tract and ovary respectively. ERalpha mRNA had the most pronounced expression in epithelial cells and subepithelial stromal cells from the oviduct to the vagina, while in the ovary it was moderately detected in only the theca folliculi and interstitial glands. The oviduct showed a region-dependent expression of ERalpha mRNA: the isthmus had the most intense signals while the infundibulum revealed a low intensity of expression. Signals for ERbeta mRNA in the ovary were most intense in the granulosa cells of healthy follicles, whereas degenerating follicles lacked any significant expression. Less intense signals for ERbeta mRNA were localized in the theca folliculi and corpus luteum. Detectable levels of ERbeta mRNA were observed in the subepithelial stromal cells from the oviduct to the vagina. This study shows that the two ER subtypes are differentially expressed in cells and compartments of the reproductive organ, suggesting that the mediation of oestrogen action in these tissues may be accomplished through the respective predominant receptor.

Animals↗

Developmental changes of the oestrogen receptor-alpha and -beta mRNAs in the female reproductive organ of the rat--an analysis by in situ hybridization.

This study employed an in situ hybridization technique to compare the cellular expression of oestrogen receptor (ER) subtypes, ER alpha and beta, in the female reproductive organ of the rat during prenatal and postnatal periods. Diffuse signals of ER alpha and beta mRNAs were co-expressed in the foetal ovary; they were weak and inconsistent before onset of gonadal differentiation, but increased in intensity with age. ER beta mRNA signals in the ovary sharply increased in intensity to adult levels by postnatal days 6-7, whereas those of ER alpha mRNA remained unchanged after birth. ER alpha was the sole subtype expressed during the prenatal period from the oviduct to the vagina, being localized mainly to the sub-epithelial stromal cells, and remained predominant thereafter. Signals for ER alpha mRNA in the epithelia were confined to the oviduct during prenatal and early postnatal periods; those in uterine and vaginal epithelia first appeared by postnatal days 4-5 and 6 respectively. Expressions of ER beta mRNA in the reproductive tract were absent during the prenatal period, and were weakly expressed during the postnatal period. Thus, oestrogen action in the developing ovary may be co-mediated by both ER alpha and beta, whereas ER alpha may be the primary mediator in the differentiation and growth of the female reproductive tract.

Animals↗

Morphology and functional roles of synoviocytes in the joint.

The joint capsule exhibits a unique cellular lining in the luminal surface of the synovial membrane. The synovial intimal cells, termed synoviocytes, are believed to be responsible for the production of synovial fluid components, for absorption from the joint cavity, and for blood/synovial fluid exchanges, but their detailed structure and function as well as pathological changes remain unclear. Two types of synoviocytes, macrophagic cells (type A cells) and fibroblast-like cells (type B cells) have been identified. Type A synoviocytes are non-fixed cells that can phagocytose actively cell debris and wastes in the joint cavity, and possess an antigen-presenting ability. These type A cells, derived from blood-borne mononuclear cells, can be considered resident macrophages (tissue macrophages) like hepatic Kupffer cells. Type B synoviocytes are characterized by the rich existence of rough endoplasmic reticulum, and dendritic processes which form a regular network in the luminal surface of the synovial membrane. Their complex three-dimensional architecture was first revealed by our recent scanning electron microscopy of macerated samples. The type B cells, which are proper synoviocytes, are involved in production of specialized matrix constituents including hyaluronan, collagens and fibronectin for the intimal interstitium and synovial fluid. The proliferative potentials of type B cells in loco are much higher than type A cells, although the transformation of subintimal fibroblasts into type B cells can not be excluded. In some mammals, type B cells show features suggesting endocrine and sensory functions, but these are not recognized in other species. The synoviocytes, which form a discontinuous cell layer, develop both fragmented basement membranes around the cells and junctional apparatus such as desmosomes and gap junctions. For an exact understanding of the mechanism of arthritis, we need to establish the morphological background of synoviocytes as well as their functions under normal conditions.

Animals↗

Immunoreactivity to a monoclonal antibody (OS-3) is shared by osteoclasts and bicarbonate-secreting cells.

OS-3, a monoclonal antibody raised against macrophagic cells derived from cultured rat glomeruli, reacts with the plasma membrane of various bicarbonate-secreting cells such as epithelial cells of the pancreatic excretory duct and type B intercalated cells of the kidney, suggesting that the antigenic molecule of OS-3 is involved in bicarbonate production and/or secretion. Since osteoclasts must vigorously extrude bicarbonate to maintain cytoplasmic pH in a physiologic range during proton secretion, we examined the localization of OS-3 immunoreactivity in the bone tissue to determine the involvement of the detected molecule in the transmembrane transport of bicarbonate in osteoclasts. The OS-3 selectively stained the basolateral plasma membrane of osteoclasts. Ultrastructurally, the immunoreactivity with OS-3 was associated with small cytoplasmic projections and microplicae of the basolateral plasma membrane. This finding suggests that osteoclasts express the molecule common to bicarbonate-secreting cells to utilize it for bicarbonate transport during bone resorption.

Animals↗

Immunohistochemical demonstration of nerve terminals in the whole hard palate of rats by use of an antiserum against protein gene product 9.5 (PGP 9.5).

Sensory innervation of the entire hard palate was investigated in the rat using serial sections immunostained for protein gene product 9.5 (PGP 9.5), a neuronal marker. PGP 9.5-immunoreactive nerve endings were widely distributed in the hard palate, but the innervation pattern and density differed among portions. They were numerous at papillary protrusions including the incisal papilla, antemolar/intermolar rugae, and postrugal filiform papillae. Immunoreactive free nerve endings gathered at the summits of the connective tissue papillae, some of them entering deeply into the epithelium. Electron microscopy demonstrated that nerves in the postrugal filiform papillae reached the stratum corneum. The atrial region, possibly the most sensitive in the hard palate, showed unique innervation: its anterior part, adjacent to incisors, developed intraepithelial networks of fine and beaded nerves, whereas its posterior part revealed cone-shaped nerve terminals formed on the connective tissue papillae of the atrial folds which comprised two lines of longitudinal flaps. Taste bud-like corpuscles gathered in the medial walls of the incisal canals and in the "Geschmacksstreifen" (taste stripes) present at the most anterior part of the soft palate. The hard palate of the rat is thus richly innervated, and is characterized by region-specific nerve endings which may be involved in mechano- and chemoreception in the oral cavity.

Animals↗