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Biomedical subjects

H Kume

Publications and source records attributed to H Kume.

At least 73 records · Page 4Linked to original sources
PubMed1998

Loss of imprinting of igf2 in renal-cell carcinomas.

Loss of imprinting (LOI) of the igf2 and h19 genes has been found not only in embryonal tumors but also in common adult cancers. To determine any possible role of genomic imprinting in the development of renal-cell carcinomas (RCCs), we examined the imprinting status of igf2 and h19 in a series of 22 such tumors, and studied its relation to their mRNA expression. Of 14 RCC specimens heterozygous for the ApaI polypmorphism, 7 (50%) showed LOI of igf2. In contrast, for h19 all 9 informative cases maintained imprinting. Furthermore, all 7 cases with LOI transcribed igf2 mRNA at elevated levels, while H19 expression was low regardless of the imprinting status compared with that of background level in each case. These results suggest that LOI of igf2, but not of h19, plays a role in the case of human RCC. However, in contrast to that in Wilms' tumor, LOI in RCC was not associated with any specific down-regulation of h19.

Adult↗
PubMed1998

Stimulation of rat hepatocyte proliferation in vitro and in vivo by factors derived from the bovine small intestinal mucosa.

A factor with a molecular weight of less than 1 kDa in the mucosa of the bovine small intestine (low molecular weight factor or LMW factor) stimulated DNA synthesis in rat hepatocytes in primary culture. This factor only showed its activity when it was added with a larger factor with a molecular weight of 30 kDa that was also found in the same tissue (high molecular weight factor or HMW factor). The LMW factor probably acts to enhance the action of a hepatotrophic growth factor, since EGF and HGF can substitute for the HMW factor. The action of the LMW factor was not due to the actions of low molecular weight substances such as norepinephrine, estradiol, triiodothyronine, and putrescine, which enhance the action of EGF or HGF, since substantial amounts of these substances were not found in the extract. When intraperitoneally administered into rats, after two-thirds hepatectomy, the LMW factor enhanced hepatocyte proliferation without the administration of the HMW factor. In the regenerating liver, a hepatotrophic growth factor(s), which acts synergistically with the LMW factor, might be properly provided, but the supply of the LMW factor might be below the level that maximally stimulates hepatocyte proliferation.

Animals↗
PubMed1998

Neutral endopeptidase 3.4.24.11 inhibition potentiates the inhibitory effects of type-C natriuretic peptide on leukotriene D4-induced airway changes.

1. Microvascular leakage, a primary feature of inflammation, is well known for worsening the asthmatic condition. Gene expression of and a specific receptor for type-C natriuretic peptide (CNP), initially considered a neuropeptide, have been detected in the human vascular wall and secretion of CNP from vascular endothelial cells has recently been demonstrated. These facts suggest the presence of a vascular natriuretic peptide system and led us to expect that CNP may act beneficially on airway microvascular leakage in asthma. In the present study, we investigated the effects of CNP against leukotriene (LT) D4-induced airway microvascular leakage and bronchoconstriction and how these effects were potentiated by thiorphan, a potent neutral endopeptidase 3.4.24.11 (NEP) inhibitor. 2. Anaesthetized male guinea-pigs, ventilated via a tracheal cannula, were placed into a plethysmograph for 10 min, in order to measure pulmonary mechanics and mean blood pressure, after challenge with 2 micrograms/kg LTD4 and then the extravasation of 20 mg/kg Evans blue dye into airway tissue was investigated to indicate and evaluate microvascular leakage. 3. Intravenous administration of CNP (100, 300 and 1000 micrograms/kg) significantly inhibited the LTD4-induced microvascular leakage and bronchoconstriction in a dose-dependent manner. These inhibitory effects were enhanced by pretreatment with 20 mg/kg thiorphan, suggesting the important role of NEP in the pulmonary metabolism of CNP. 4. We believe that these results are encouraging for the further investigation of the therapeutic applications of exogenous CNP in asthma.

Animals↗
PubMed1998

Phosphodiesterase IV inhibitors synergistically potentiate relaxation induced by forskolin in guinea-pig trachea.

1. Beta-adrenoceptor receptor agonists are the principal bronchodilator agents used in the treatment of bronchial asthma. However, the regular use of beta-adrenoceptor agonists in asthmatic patients is likely to increase asthma severity because of a defective beta-adrenoceptor function. Bronchodilators that bypass this defective function are therefore needed. 2. Our objectives in this study were: (i) to assess the effects of an agent that directly activates adenylate cyclase (forskolin) on guinea-pig tracheal smooth muscle; (ii) to study the interactions between selective cyclic nucleotide phosphodiesterase (PDE) inhibitors and forskolin by measuring isometric tension; and (iii) to compare these results with the interaction between PDE inhibitors and terbutaline, a beta-adrenoceptor agonist. 3. The relaxant effects of forskolin alone, which is now under development as a new bronchodilator for bronchial asthma therapy, were slightly weaker than those of terbutaline on guinea-pig tracheal smooth muscle. 4. Both denbufylline and Ro 20-1724, cyclic nucleotide PDE IV inhibitors, synergistically increased the relaxant effects of forskolin and terbutaline, while other PDE isozyme inhibitors (amrinone, vinpocetine and zaprinast) had only a minor influence. 5. In conclusion, a good synergistic interaction between forskolin and PDE IV inhibitors, especially denbufylline, may provide a means for bypassing beta-adrenoceptors. Thus, the combination of forskolin and PDE inhibitors would become useful in the treatment of bronchial asthma.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1998

Evaluation of antifungal activity of an antifungal drug by in vitro simulation of in vivo pharmacokinetics of the drug against fungal hyphal growth.

An automatic drug concentration simulator (DCS) has been developed and its applicability has been demonstrated by in vitro simulation of the human plasma concentration-time curve of fluconazole (FLCZ) against hyphal growth of Candida albicans and Aspergillus fumigatus. The response of hyphal growth to FLCZ was continually monitored and analyzed using an automatic hyphal growth analyzing system (Bio-Cell Tracer). The simulated concentration of FLCZ by DCS was confirmed by HPLC. The DCS assay was reproducible with a mean coefficient of variation (C.V., n=3) of 5.38 %. When the growth of C. albicans hyphae was tested, there was a lag of onset of FLCZ effect between the time when FLCZ concentration became maximal (C MAX, 7.95 microg/ml ) and the point at which hyphal growth ceased. In contrast, FLCZ was found inactive against A. fumigatus. The newly devised technique could provide clinicians with important information in determining optimal dosing regimens for antifungal drugs.

Antifungal Agents↗
PubMed1998

The presenilin 2 loop domain interacts with the mu-calpain C-terminal region.

Presenilin 2 (PS2) is a gene responsible for the early-onset familial Alzheimer's disease (AD). PS2 mutations are considered to be closely related to the pathogenesis of AD. We screened for proteins that interact with PS2 to understand its pathological and physiological functions. Using the PS2 loop domain as the bait, the yeast two-hybrid system was used for screening, and mu-calpain was identified as a PS2 binding protein. In COS-1 cells, the interaction of PS2 with mu-calpain was confirmed by immunoprecipitation. These results suggested that PS2 and mu-calpain interact with each other, and might regulate each other's functions.

Animals↗
PubMed1998

Pityriasis versicolor with a unique clinical appearance.

We experienced an atypical case of pityriasis versicolor with a unique clinical appearance and undescribed mycological features. Although Malassezia sp. was cultured from the keratotic material, the fungal elements observed in the material were not readily identified as Malassezia. The diagnosis was established with the aid of immunohistochemical and ultrastructural studies with the aetiological agent being identified as M. globosa.

Diagnosis, Differential↗
PubMed1998

Effects of atrial natriuretic peptide and 8-brom cyclic guanosine monophosphate on human tracheal smooth muscle.

The relaxant effects of intracellular concentration of cyclic guanosine monophosphate (cGMP) on spontaneous tone in human tracheal smooth muscle were investigated in comparison with guinea pig, using isometric tension records. In both human and guinea pig tracheas, application of atrial natriuretic peptide (ANP) and 8-brom cGMP (a membrane permeable analogue of cGMP) caused an inhibition of spontaneous tone in a concentration-dependent fashion. However, ANP was less potent in relaxation of tracheal smooth muscle in human than guinea pigs, and values of % relaxation induced by 1 mmol/l ANP in human and guinea pigs were 37.1 +/- 5.3 and 82.7 +/- 10.5%, respectively (n = 6). In the presence of 30 nmol/l iberiotoxin (IbTX), a potent and selective large conductance Ca(2+)-activated K+ (BKCa) channel inhibitor, relaxant actions of ANP on human tracheal smooth muscle were markedly suppressed, and values of % relaxation by 1 mmol/l ANP decreased to 8.4 +/- 1.2% (n = 6). On the other hand, 8-brom cGMP was roughly equipotent in relaxating tracheal smooth muscle in these two species, different from ANP, and inhibitory effects of 8-brom cGMP on both human and guinea pig tracheal smooth muscle were also markedly suppressed in the presence of 30 nmol/l IbTX, similar to ANP. These results demonstrate that augmentation of BKCa channel activity may play a functionally important role in the cGMP-induced relaxation in human airway smooth muscle. However, ANP may have modest potency as a bronchodilator.

Aged↗
PubMed1997

A novel brain gene, norbin, induced by treatment of tetraethylammonium in rat hippocampal slice and accompanied with neurite-outgrowth in neuro 2a cells.

Tetraethylammonium (TEA) induces long-term potentiation (LTP)-like synaptic enhancement in rat hippocampal slices. To find the genes related to this phenomenon, subtraction screening was performed between the mRNA of TEA-treated slices and that of untreated whole brain. One of the clones induced by the TEA treatment, named as norbin, was expressed only in neural tissues. The predicted protein sequence of norbin consisted of 729 amino acids, and no homologies in the sequence were found with known genes or proteins. Overexpression of norbin in cultured Neuro 2a cells by cDNA transfection induced neurite-outgrowth. Since in the course of neural plasticity the formation of new synapses should occur, the neurite-outgrowth-related protein, norbin, might play an important role in neural plasticity.

Amino Acid Sequence↗
PubMed1997

Ethanolamine modulates the rate of rat hepatocyte proliferation in vitro and in vivo.

A low molecular weight, heat-resistant hepatotrophic factor in an extract from the bovine intestinal mucosa was purified and identified as ethanolamine by structural analyses. The mode of action of ethanolamine in vitro and in vivo coincided with that of the crude extract of the tissue, indicating that ethanolamine is the active component. Ethanolamine synergistically elevated the stimulation of DNA synthesis in hepatocytes in primary culture when added together with a growth factor, such as epidermal growth factor, with the ED50 being 20 microM, although it showed little stimulatory effect by itself. Contrary to these in vitro results, the intraperitoneal administration of ethanolamine hydrochloride (24 mg of ethanolamine per kg of body weight) enhanced hepatocyte proliferation in regenerating rat livers after two-thirds hepatectomy without the administration of any growth factors. In the regenerating liver, hepatocyte proliferation may be initiated by an endogenous growth factor, but the supply of ethanolamine in circulation may not be sufficient for optimal hepatocyte proliferation; thus, the exogenous administration of ethanolamine may further enhance hepatocyte proliferation. Ethanolamine in circulation may be a humoral hepatotrophic factor.

Animals↗
PubMed1997

Cleft palate and decreased brain gamma-aminobutyric acid in mice lacking the 67-kDa isoform of glutamic acid decarboxylase.

In addition to its role as an inhibitory neurotransmitter, gamma-aminobutyric acid (GABA) is presumed to be involved in the development and plasticity of the nervous system. GABA is synthesized by glutamic acid decarboxylase (GAD), but the respective roles of its two isoforms (GAD65 and 67) have not been determined. The selective elimination of each GAD isoform by gene targeting is expected to clarify these issues. Recently we have produced GAD65 -/- mice and demonstrated that lack of GAD65 does not change brain GABA contents or animal behavior, except for a slight increase in susceptibility to seizures. Here we report the production of GAD67 -/- mice. These mice were born at the expected frequency but died of severe cleft palate during the first morning after birth. GAD activities and GABA contents were reduced to 20% and 7%, respectively, in the cerebral cortex of the newborn GAD67 -/- mice. Their brain, however, did not show any discernible defects. Previous pharmacological and genetic investigations have suggested the involvement of GABA in palate formation, but this is the first demonstration of a role for GAD67-derived GABA in the development of nonneural tissue.

Aging↗
PubMed1997

The presenilin 2 mutation (N141I) linked to familial Alzheimer disease (Volga German families) increases the secretion of amyloid beta protein ending at the 42nd (or 43rd) residue.

To gain insights into the significance of presenilins (PS) in the pathogenetic mechanisms of early-onset familial Alzheimer disease (FAD), we expressed cDNAs for wild-type PS2 and PS2 with the Volga German (N141I) mutation in cultured cells and then examined the metabolism of the transfected proteins and their effect on the C-terminal properties of secreted amyloid beta protein (A beta). PS2 was identified as a 50- to 55-kDa protein, which was cleaved to produce N-terminal fragments of 35-40 kDa and C-terminal fragments of 19-23 kDa. The Volga German (N141I) mutation did not cause any significant change in the metabolism of PS2. COS-1 cells doubly transfected with cDNAs for N141I mutant PS2 and human beta-amyloid precursor protein (betaAPP) or a C-terminal fragment thereof, as well as mouse Neuro2a neuroblastoma cells stably transfected with N141I mutant PS2 alone, secreted 1.5- to 10-fold more A beta ending at residues 42 (or 43) [A beta42(43)] compared with those expressing the wild-type PS2. These results strongly suggest that the PS2 mutation (N141I) linked to FAD alters the metabolism of A beta/betaAPP to foster the production of the form of A beta that most readily deposits in amyloid plaques. Thus, mutant PS2 may lead to AD by altering the metabolism of A beta/betaAPP.

Alzheimer Disease↗
PubMed1997

Acetylcholine-induced vasoconstrictor response of coronary vessels in rats: a possible contribution of M2 muscarinic receptor activation.

A mechanism by which acetylcholine (ACh) may elicit vasoconstrictor response in coronary vessels was studied in rat hearts perfused at a constant flow rate. In spontaneously beating hearts, bolus injections of ACh and carbachol (CCh) produced biphasic changes in coronary perfusion pressure (CPP): a transient increase at the initial period followed by a sustained decrease. In KCl-arrested hearts, ACh and CCh produced a monophasic increase in CPP, which was attenuated by either removal of endothelial cells by saponin or cyclooxygenase inhibition by diclofenac sodium. In the spontaneously beating heart, ACh-induced vasoconstriction was almost abolished by atropine (0.1 microM) and was markedly attenuated by an M2 antagonist, methoctramine (0.1 microM), but not by an M1 antagonist, pirenzepine (1 microM). Arecaidine propargyl ester (APE), an M2 agonist, produced coronary artery constriction which was attenuated by methoctramine (0.1 microM) but not by pirenzepine (0.1 microM) in both spontaneously beating and KCl-arrested hearts. McN-A-343, an M1 agonist, increased CPP in both beating and KCl-arrested hearts, but to a lesser degree than APE. These results suggest that the release of vasoconstrictor prostaglandins from endothelial cells contributes to the vasoconstrictor response to ACh in perfused rat coronary vessels, and the response to ACh appears to be mediated, in part, via the M2 subtype of muscarinic receptors.

Acetylcholine↗
PubMed1997

Effects of BKCa channels on the reduction of cytosolic Ca2+ in cGMP-induced relaxation of guinea-pig trachea.

1. In order to examine the mechanisms of cGMP-induced relaxation in airway smooth muscle, the effects of atrial natriuretic peptide (ANP) and 8-brom cGMP on muscle tone were studied by measuring isometric tension, while the effects on cytosolic Ca2+ concentrations were studied by measuring the spectra of fura-2 loaded in guinea-pig tracheal strips. 2. Atrial natriuretic peptide and 8-brom cGMP caused a concentration-dependent inhibition of spontaneous tone in the guinea-pig trachea. The relaxant effects of these agents on spontaneous tone were markedly suppressed in the presence of iberiotoxin (IbTX), a selective inhibitor of large-conductance Ca2(+)-activated K+ (BKCa) channels. Iberiotoxin (30 nmol/L) markedly affected the maximal effect induced by ANP and 8-brom cGMP and augmented EC70 values for ANP and EC50 values for 8-brom cGMP approximately 27- and 17-fold, respectively. The inhibitory effects of IbTX on relaxation induced by these agents were diminished in the presence of 1 mumol/L nifedipine, an antagonist of voltage-operated Ca2+ channels (VOCC). 3. The inhibitory action of ANP and 8-brom cGMP on spontaneous tone was not affected by the presence of 10 mumol/L glibenclamide, an inhibitor of ATP-sensitive K+ channels, and 100 nmol/L apamin, an inhibitor of small-conductance Ca2(+)-activated K+ channels. When these agents were applied to tissues precontracted by high (40 mmol/L) K+, the relaxant effects of these agents markedly diminished. 4. The extracellular Ca2(+)-dependent contraction was inhibited in the presence of 0.3 mumol/L ANP or 0.1 mmol/L 8-brom cGMP. Concentration-response curves to extracellular Ca2+ (0.03-2.4 mmol/L) were markedly diminished by exposure to these agents. The maximal effect induced by extracellular Ca2+ was affected by these agents. 5. Atrial natriuretic peptide caused an inhibition of spontaneous tone accompanied by a reduction in the intracellular Ca2+ concentration. In the presence of IbTX, the elimination of both muscle tone and cytosolic Ca2+ by ANP was suppressed. 6. We conclude that ANP and 8-brom cGMP activate BKCa channels and that the inhibition of Ca2+ influx through VOCC, mediated by BKCa channel activation, may be involved in cGMP-dependent bronchodilation.

Animals↗
PubMed1997

Inhibitory effects of Gs on desensitization of beta-adrenergic receptors in tracheal smooth muscle.

We examined the reduction of mechanical responsiveness to beta-agonists after continuous or repeated exposure to agonists in guinea pig tracheal smooth muscle, using an isometric tension record. The inhibitory action of 0.1 microM isoproterenol (Iso) on contraction induced by 1 microM methacholine (MCh) was suppressed markedly after incubation with 0.0003-3 microM Iso for 1 h. The effects of Iso on MCh-induced contraction gradually decreased after repeated application of these agents at intervals of 1 h. When Iso was perfused cumulatively to tissues repeatedly precontracted by MCh, the effects of Iso were reduced at the second application. This reduced responsiveness to beta-agonists was not mimicked by forskolin and did not occur after preexposure of the strips to 2 micrograms/ml cholera toxin for 6 h. We concluded that this reduced responsiveness to beta-agonists is homologous desensitization and that the irreversible activation of the stimulatory G proteins, Gs, may play an important role in prevention of the reduced responsiveness to beta-agonists.

Adrenergic beta-Agonists↗
PubMed1997

[Significance of transrectal ultrasound and sextant systematic core biopsy for performing radical prostatectomy].

BACKGROUND: To estimate the usefulness of sextant systematic core biopsy or transrectal ultrasonography (TURS) for performing radical prostatectomy. METHODS: The findings of sextant biopsy and TRUS were compared with 52 step-sectioned specimens obtained from radical prostatectomy. RESULTS: In 34 cases with no influence of hormonal therapy at the time of TRUS and biopsy, sextant systematic core biopsy provided tumor distribution rather precisely. In 33% of the cases who had received hormonal therapy, tumor cells were not detected by this sextant biopsy series. In these cases, majority of residual cancer existed in transition zone, paraurethral or fibromuscular stroma. Six cases showed small adenocarcinoma in only one biopsy tip obtained from sextant biopsy, while 4 cases were revealed well differentiated adenocarcinoma (Gleason score less than 4) by these core biopsies. Comparing with tumor mapping, Gleason score, PSA level and pT stage of the radical prostatectomy specimens, these tumors presented as, not clinically insignificant, but clinically significant prostate cancer. Playing special attention to distraction of normal ultrasound zonal configuration, TRUS detected neurovascular invasion with 94.7% sensitivity, 78.3% positive predictive value and 90. 9% negative predictive value, while seminal vesicle invasion with 75% sensitivity, 50% positive predictive value, 90.9% negative value. CONCLUSION: Sextant biopsy tended to underestimate the tumors located in the transition zone, paraurethral and fibromuscular lesion. Additional or direct biopsies in transition zone are indispensable for accurate diagnosis. Findings of TRUS and distribution of positive core biopsy from sextant biopsy enable to extract stage C prostate cancer providing negative surgical margin.

Biopsy↗
PubMed1997

[Retrograde radical cystectomy. Advantages of our "vesico-rectal tunnel" method].

BACKGROUND: The facilitation of dissecting the vesical pedicles and undisturbed preservation of the membranous urethra were investigated during radical cystectomy using the retrograde technique. METHODS: First, just as with radical prostatectomy, the prostate and the rectum were separated by blunt digital dissection. After the deep dorsal vein complex and the urethra were cut, the bladder and the rectum were also separated in a retrograde manner towards the Denonvillier's fascia. Then the peritoneum was opened and its lowest part was incised above the cul-de-sac. A tunnel was made beginning at the cut-end of the urethra to the cul-de-sac. Consequently, the bladder was lifted up by hand inserted this "vesico-rectal tunnel" and the bilateral remaining lateral pedicles were ligated and transsected without difficulty. RESULTS: Fourteen patients underwent radical cystectomy using this technique. Among them, the average operating time and blood loss in 4 patients received retrograde radical cystectomy accompanied with ileal conduit were 5 hours 15 minutes and 1606 ml, respectively. These in 9 patients received retrograde radical cystectomy followed by bowel orthotopic urinary reservoir were 7 hours and 6 minutes and 1086 ml, respectively. Another patient received ureterocutaneoustomy. CONCLUSION: Creating a "vesico-rectal tunnel" during radical cystectomy primarily by the retrograde extraperitoneal technique can afford to preserve the urethral sphincter and to ligate the pedicles easily. This method is fundamentally familiar to us because retrograde radical prostatectomy is now widely adopted and it may help to reduce the operating time even when there is a shortage of manpower.

Aged↗