Prevention of reperfusion injury after rat pancreas preservation using rinse solution containing nafamostat mesilate.
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Adenosine↗
Biomedical subjects
M Nishihara
Publications and source records attributed to M Nishihara.
Effect of donor fasting on survival of pancreas and heart grafts after warm ischemia.
Livers from fasted animals are believed to be more vulnerable to ischemic injury than those from fed donors. However, we have recently shown the opposite: livers from fasted rats were more tolerant to ischemic injury. Indeed, the survival rate of 60 min warm ischemic damaged livers increased from 0 to 90% if donor rats were fasted for three days. In this study, we examined how donor fasting affects the outcome of pancreas and heart preservation. BN rats were used as both donors and recipients, and recipients of pancreatic grafts were rendered diabetic prior to transplantation. Pancreatic or heart grafts were subjected to 90 min or 25 min of warm ischemia and were transplanted into the right side of the necks of recipients rats. The viability rate of hearts transplanted from fed donors into fed recipients was only about 11% (1/9) after transplantation. However, the viability rate with fasted donors was 75% (6/8). The rate of successful pancreatic grafting from fed donors into fed recipients was 28.6% (2/7), and that from fasted donors to fed recipients was 41.7% (5/12). These results confirm that the nutritional status of the donor is an important factor in the outcome of not only liver, but also pancreas and heart preservation during transplantation, although the effect of fasting on pancreatic graft is marginal.
Animals↗
Excitability of neurons in the ventromedial nucleus in rat hypothalamic slices: modulation by amino acids at cerebrospinal fluid levels.
Free amino acid composition of cerebrospinal fluid (CSF) in the rat was measured to examine if electrical activity of neurons in the ventromedial nucleus of the hypothalamus (VMH) is modulated by amino acids at CSF levels. In CSF collected through a cannula in the cerebromedullary cistern of freely moving rats, the total concentration of amino acids was 979 microM, while that in plasma was 3.50 mM. Glutamine was the major (60%) component and each of other amino acids was less than 8.0% (lysine) of total. Among neutral amino acids, concentrations of serine, threonine and alanine were much higher than those of others. Although concentrations of basic amino acids such as lysine and arginine were also relatively high, those of acidic amino acids such as glutamate and aspartate were very low. Spontaneous firing rates of 73% of the neurons in the VMH in brain slices in vitro were facilitated by changing the perfusion medium from artificial CSF (ACSF) without amino acids to one containing each amino acid close to CSF levels (AA-ACSF). Glutamine (600 microM) alone similarly facilitated the activity in 64% of VMH neurons, though the facilitatory effect of amino acids was blocked by simultaneous application of neither AP-5 nor CNQX, antagonists for NMDA or non-NMDA receptors, respectively, to AA-ACSF at 100 microM.(ABSTRACT TRUNCATED AT 250 WORDS)
Action Potentials↗
Two new phospholipids, hydroxyarchaetidylglycerol and hydroxyarchaetidylethanolamine, from the Archaea Methanosarcina barkeri.
The structures of two new ether phospholipids of the methanogenic Archaea, Methanosarcina barkeri, were determined as hydroxyarchaetidylglycerol and hydroxyarchaetidylethanolamine by means of chemical, chromatographic and enzymatic analyses, and fast atom bombardment-mass spectrometry. These lipids are hydroxy diether analogs of phosphatidylglycerol and phosphatidylethanolamine, respectively, with beta-hydroxyarachaeol (2-O-(3'-hydroxy)phytanyl-3-O-phytanyl-sn-glycerol) as a core lipid. In addition, two other ether phospholipids, usual archaetidylglycerol and archaetidylethanolamine, were also identified in the organism. The stereochemical structure of the unalkylated glycerophosphate of hydroxyarchaetidylglycerol and archaetidylglycerol was determined as sn-glycerol-3-phosphate by use of sn-glycerol-3-phosphate dehydrogenase. The stereochemical configuration of the glycerophosphoglycerol backbone of these lipids was a mirror image of that of diacylphosphatidylglycerol from the organisms of the domains Bacteria and Eucarya, and it was shared with extremely halophilic Archaea. These four phospholipids, in addition to five lipids that had already been reported, accounted for 88% of the total polar lipids of this organism.
Chromatography, Gas↗
Promoter analysis of seed storage protein genes from Canavalia gladiata D.C.
A number of A/T-rich sequences and a CATGCAT/A sequence are contained in the 5'-upstream regions of the genes encoding concanavalin A (Con A) and canavalin, two major seed storage proteins of Canavalia gladiata D.C. To study the role of these sequences in the seed-specific gene expression, we constructed 5'-deletion mutants and examined the transient expression of beta-glucuronidase reporter gene by particle bombardment and the stable expression by Agrobacterium-mediated transformation of tobacco plants. Positive regulatory elements were located in the -894/-602 and -602/-74 regions of the Con A gene, and in the -428/-376, -281/-155 and -155/-50 regions of the canavalin gene. In addition, the results suggested that the A/T-rich sequences in the 5'-upstream region of the Con A gene play a role in transcriptional activation, but that those of the canavalin gene have little effect on the gene expression. The CATGCAT/A sequence was not sufficient by itself for high levels of expression of both the Con A and canavalin genes. The canavalin polypeptide amounted to about 1% of the total extractable protein in the transgenic tobacco seeds, but the Con A polypeptide was not detected in the extractable protein.
Base Sequence↗
Successful expression in pollen of various plant species of in vitro synthesized mRNA introduced by particle bombardment.
Gold particles coated with beta-glucuronidase (GUS) mRNA with a 5' cap structure that had been synthesized in vitro were introduced, by use of a pneumatic particle gun, into pollen grains of lily (Lilium longiflorum), freesia (Freesia refracta) and tulip (Tulipa gesneriana). A fluorometric assay for the GUS activity indicated that in vitro synthesized GUS mRNA introduced into these pollen cells by particle bombardment was successfully expressed. GUS activity in extracts of the bombarded lily pollen became detectable fluorometrically within 30 min after bombardment, peaked at 6 h, then gradually decreased. This activity changed as a function of the developmental stage of the pollen cell of lily.
Gene Expression↗
Peculiar enlargement of the nasopharynx in patients with anorexia nervosa.
We examined the nasopharynx and brain in 17 patients with anorexia nervosa by CT and compared the findings with those of 44 normal subjects and of 5 patients of the same age with marked emaciation caused by various psychiatric disorders. An enlarged nasopharyngeal space with a flattened posterior wall and enlarged lateral pharyngeal recesses were demonstrated in all patients with anorexia nervosa whose weight was lowest at the time of the CT examination, and these CT features regressed or became normal quickly after they had gained some weight. This characteristic enlargement of the nasopharynx and lateral pharyngeal recesses was observed neither in the markedly emaciated patients (2 with schizophrenia, 1 with major depression, 1 with stupor and the other with an extremely unbalanced diet) nor in 44 normal subjects without emaciation. These features were therefore thought to be characteristic and of diagnostic significance.
Adolescent↗
Synthesis of type III collagen and type IV collagen by tubular epithelial cells in diabetic nephropathy.
Overproduction of extracellular matrix (ECM) is considered to be primarily responsible for both glomerular and tubulointerstitial (TI) changes in diabetic nephropathy (DN). To clarify the possible role of the collagens in TI damage in DN, type III interstitial collagen and type IV basement membrane collagen were studied in 10 cases of DN and 10 control cases by immunohistochemistry and in situ hybridization techniques. In control cases, no immunostaining for type III collagen was found in the renal tubules, while strongly positive in the adjacent interstitium. On the other hand, type IV collagen was found weakly in the tubular basement membrane (TBM) in control cases. In DN, increased immunostaining for both type III and type IV collagens were found in the damaged tubulointenstitium (TI). To determine the sources of these collagens in TI damage, non-radioactive in situ hybridization was performed utilizing thymine-thymine (T-T) dimerized synthetic oligonucleotides complementary to either human pro alpha 1 (III) chain or pro alpha 1 (IV) chain mRNA as probe. In normal tubules, tubular epithelial cells were not uniformly but persistently positive for pro alpha 1 (IV) mRNA. Meanwhile, no specifically detectable positive hybridization signals for pro alpha 1 (III) mRNA was found in the normal tubular epithelial cells. Accelerated synthesis of both type III and type IV collagens by tubular epithelial cells was noted in TI damage in DN. From the results we concluded that excessive synthesis of both type III and type IV collagens by tubular epithelial cells might significantly contribute to the TI damage found in DN.
Collagen↗
sn-glycerol-1-phosphate dehydrogenase in Methanobacterium thermoautotrophicum: key enzyme in biosynthesis of the enantiomeric glycerophosphate backbone of ether phospholipids of archaebacteria.
One of the most characteristic features of archaebacterial ether phospholipids is the enantiomeric configuration of their glycerophosphate backbone (sn-glycerol-1-phosphate), that is the mirror image of the structure of the eubacterial or eukaryotic counterpart. The enzyme that forms glycerophosphate of this configuration was found for the first time in a cell-free extract of the methanogen, Methanobacterium thermoautotrophicum, and was identified as sn-glycerol-1-phosphate:NAD+ oxidoreductase (sn-glycerol-1-phosphate dehydrogenase) after partial purification. Because sn-glycerol-1-phosphate has been found to be a precursor of ether lipids of this organism, sn-glycerol-1-phosphate dehydrogenase is a key enzyme in the biosynthesis of the enantiomeric ether lipids of methanogens.
Animals↗
Elevation of plasma noradrenaline levels in urethane-anaesthetized rats by activation of central prostanoid EP3 receptors.
1. We studied the effects of intracerebroventricular (i.c.v.) administration of prostaglandin E2 (PGE2) and its receptor subtype ligands on plasma levels of catecholamines in urethane-anaesthetized rats. 2. Administration of PGE2 (0.15, 0.3 and 1.5 nmol per animal, i.c.v.) dose-dependently elevated plasma levels of noradrenaline (NA), while the levels of adrenaline were not affected. 3. Administration of sulprostone (EP3/EP1 agonist) and misoprostol (EP3/EP2 agonist) effectively elevated plasma NA levels in a dose-dependent manner (0.1, 0.3, and 1.0 nmol per animal). Butaprost (EP2 agonist) (0.3, 1.0 and 3.0 nmol per animal) was without effect. 17-Phenyl-omega-trinor PGE2 (EP1/EP3 agonist) effectively elevated plasma NA levels only at its highest dose (1.0 nmol per animal), but this elevation was not attenuated by pretreatment with SC-19220 (selective EP1 antagonist) (20 nmol per animal, i.c.v.). 4. The potency of these test agents in elevating plasma levels of NA was as follows; misoprostol > sulprostone > PGE2 > > 17-phenyl-omega-trinor PGE2 > > > butaprost. These results suggest that activation of central prostanoid EP3-receptors induces central sympathetic outflow in rats.
Abortifacient Agents, Nonsteroidal↗
Molecular cloning of cDNA for equine ovarian inhibin/activin beta A subunit.
cDNAs encoding equine inhibin/activin beta A subunit were isolated from an equine follicle cDNA library and characterized. Using primers based on the rat inhibin/activin beta A subunit cDNA sequence, a RT-PCR was performed to generate the probe for screening. Four positive clones were isolated. Analysis of the nucleotide sequence of these clones revealed that two pairs of identical clones were present, Eq-beta A-1 (0.9 kb) and Eq-beta A-2 (1.5 kb). Eq-beta A-2 clone contained a complete open reading frame encoding 426 amino acids. The deduced amino acid sequence of equine inhibin/activin beta A subunit showed high similarity (> 90%) to those of five other mammalian species. Northern blot analysis revealed that placenta from mare on day 180 of pregnancy contained a 1.5 kb inhibin/activin beta A subunit mRNA.
Activins↗
Molecular cloning of DNA for inhibin alpha-subunit from equine ovary.
cDNA encoding equine inhibin alpha-subunit precursor protein was isolated from an equine ovarian cDNA library. For screening, the DNA probe was amplified by the RT-PCR using primers designed based on the rat inhibin alpha-subunit cDNA sequence. Out of 1.2 x 10(5) plaques screened, 19 positive clones were isolated, and one of these clones (Eq-alpha-11) contained a complete open reading frame encoding 367 amino acids. The similarity of the deduced amino acid sequences of both equine inhibin alpha-subunit precursor protein and the mature protein were greater than 80% to those of other six mammalian species, including bovine, porcine, ovine, human, rat and mouse. Northern blot analysis revealed that a single major band at 1.5 kb was present in the RNA samples both from equine follicles and testes.
Amino Acid Sequence↗
Hypothalamic gonadotropin-releasing hormone gene expression during rat estrous cycle.
Expression of gonadotropin-releasing hormone (GnRH) gene in the hypothalamus of female rats was studied by the quantitative reverse transcription-polymerase chain reaction (RT-PCR) method. During the estrous cycle, the GnRH mRNA level did not change from diestrus I to II, and then increased with a peak at 1100 h on proestrus. In the afternoon of proestrus, GnRH mRNA decreased rapidly with a nadir at 1600 h, and thereafter increased again and reached a peak at 1100 h on estrus. Since hypothalamic GnRH mRNA was found to be increased by subcutaneous implantation of estradiol-containing silastic tubing in ovariectomized rats, the peak of GnRH gene expression in the proestrous morning might be due to an increase in circulating estrogen in this phase of the estrous cycle. The surge of luteinizing hormone in the proestrous afternoon and the subsequent increase in GnRH mRNA were completely blocked by the injection of MK801, an antagonist for NMDA receptors, suggesting that the excitation of GnRH neurons leads to an increase in GnRH gene expression. This was further supported by the in vitro observation that high K(+)-induced membrane depolarization markedly increased GnRH mRNA in hypothalamic slices. This increase in GnRH mRNA due to neuronal excitation does not seem to require newly synthesized proteins because anisomycin, a protein synthesis inhibitor at the level of translation, did not affect GnRH gene expression. These results suggest that the hypothalamic GnRH mRNA level reached two peaks during the rat estrous cycle, i.e., in proestrous morning and estrous morning, and that estrogen and GnRH neuronal excitation play pivotal roles in regulating GnRH gene expression.
Animals↗
Prolonged survival of hamster-to-rat pancreas xenografts by FK 506 and splenectomy.
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Animals↗
Induction of unresponsiveness to islet allografts with donor spleen cell inoculation followed by a single injection of FK 506.
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Animals↗
Participation of donor adhesion molecules in islet xenograft rejection.
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Animals↗
Potent immunosuppressive effect of anti-LFA-1 monoclonal antibody on islet allograft rejection.
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Animals↗
Concomitant regulation of running activity and metabolic change by the ventromedial nucleus of the hypothalamus.
The aim of this study was to elucidate the involvement of kainate (KA) type glutaminergic, GABAergic and adrenergic receptors in the ventromedial nucleus of the hypothalamus (VMH) in inducing running activity and metabolic adaptations. Injection of either KA or bicuculline methiodide (BM), a GABAA receptor antagonist, into the VMH of conscious rats resulted in an increase in plasma glucose, norepinephrine, epinephrine and corticosterone, as well as running activity. KA or BM increased plasma glucose and catecholamine even under urethane anesthesia. Co-injection of either alpha- or beta-adrenergic receptor antagonist, i.e. phentolamine or timolol, respectively, with KA into the VMH of conscious rats elicited only a slight increase in plasma glucose and catecholamines, though it successfully induced hyper-running. However, plasma corticosterone was higher in the animals injected with adrenergic blockers, suggesting that an insufficient supply of energy substrates would enhance the activity of the hypothalamo-pituitary-adrenal system. We conclude that: (1) KA type glutaminergic and GABAergic receptors in the VMH are involved in regulating running activity and the sympathetic nervous system; (2) the brain noradrenergic system may mediate the KA action on the sympathetic nervous system.
Animals↗