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Biomedical subjects

H Wakabayashi

Publications and source records attributed to H Wakabayashi.

At least 91 records · Page 5Linked to original sources

Effect of arachidonate on lipid metabolism in ethanol-treated rats fed with lard.

Two groups of rats, ethanol-treated and sucrose-administered control rats, were fed diets with different AA content (0, 2 and 3% weight) for 14 days. Ethanol was administered by gavage at a single daily dose of 3 g/kg body weight. The ethanol-treated rats showed significantly higher levels (p < 0.01) of serum ALT activity. The dietary AA supplement lowered the serum ALT activity and liver triglyceride both in control and ethanol-treated rats. Significantly lower levels of 20:4n - 6 and 20:4n - 6/18:2n - 6 ratio and higher levels of 18:1n - 9 in both the serum and liver triglyceride were observed in the ethanol-treated rats. The AA-supplemented diet induced a marked increase of 20:4n - 6 and subsequent significant decrease of 18:2n - 6 both in the liver and serum phospholipid in control and ethanol-treated rats. 18:1n - 9 in the serum and liver triglyceride in both groups was also markedly decreased by AA supplement. No significant difference was observed in the liver 6-keto-PGF(1 alpha) level between the ethanol-treated and control rats. In the ethanol-treated rats, the level of 6-keto-PGF(1 alpha) was elevated in the rats fed the 3% AA-supplemented diet. Though the liver leukotriene B4 levels were increased by ethanol administration in all rats, these levels were not increased by dietary AA.

6-Ketoprostaglandin F1 alpha↗

[An epidemiological study of HGV infection in an epidemic area of HCV infection in Japan].

We examined the prevalence on HGV infection in a hepatitis C epidemic area where 27.8% of inhabitants 40 years or over were anti-HCV positive. Of the 200 HCV RNA positive inhabitants 17(8.5%) were HGV RNA positive, although one of the 100 anti-HCV negative inhabitants in the same epidemic area, showed HGV RNA positive (1.0%). It might suggest that HGV infections in this HCV infection epidemic area have occurred in relation to HCV infection. The rate of abnormal transaminase in inhabitants HGV RNA and HCV RNA both positive was not different from that in inhabitants HGV RNA negative, but HCV RNA positive. This indicates that a further study is needed about the relationship between liver damage and HGV infection.

Biomarkers↗

Measurement of the expiratory ammonia concentration and its clinical significance.

Although gaseous ammonia (NH3) can freely enter cells through the plasma membrane where NH3 is cyto(neuro)toxic, NH3 and ionic ammonia (NH4+) contents have not been studied in biological materials. We developed a new method for measurement of expiratory NH3 concentration, which may reflect blood NH3 concentrations. The method is a sensor tube type-gas assay system. Expiratory NH3 concentration in patients with chronic liver diseases increased when their blood ammonia (NH4(+)+NH3) concentrations increased above 90 micrograms/dl (normal range; 12-66 micrograms/dl). However, cirrhotic patients, who had relatively higher expiratory NH3 concentration compared to blood NH3 concentrations (calculated from Henderson-Hasselbalch formula), were found to have subclinical encephalopathy. Measurement of expiratory NH3 concentration may be of clinical significance for the diagnosis of encephalopathy associated with hyperammonemia.

Ammonia↗

Etiology and prognosis of cryptogenic liver cirrhosis: possible contribution of hepatitis B virus.

Three female patients without type B or type C viral hepatitis, alcoholic, metabolic or autoimmune liver disease, were selected from 250 cases with histologically proven liver cirrhosis (M:F = 183:67). All three cases showed at least one positive aspect among three parameters of serum anti-HBc (RPHA, x1), HBV-DNA (gene S, nested PCR) and liver HBs and/or pre-S2 antigen (immunoperoxidase methods). Two cases may suggest a spontaneous disappearance of HBV from sera. Another case may suggest a contribution of mutant HBV which can not be detected by the routine tests. These HBV-related cirrhotic patients have done well clinically and have not been associated with hepatocellular carcinoma during the period from 6 to 12 years of follow-up when compared with 59.6% and 65.4% prevalence of hepatocarcinogenesis in type B and type C hepatitis-associated cirrhosis during the observation period of six and seven years on average, respectively.

Aged↗

Autoimmune hepatitis types IIa and IV in young men.

Two young men with autoimmune hepatitis types IIa and IV are reported here. These rare types of the disease could be diagnosed by using the diagnostic criteria for scoring of autoimmune hepatitis established by the International Group.

Actins↗

Direct evidence for a role of glutamate in the expression of the opioid withdrawal syndrome.

To investigate the role of glutamate in the expression of the withdrawal signs from opioids, rats were intracerebroventricularly (i.c.v.) infused continuously with morphine (a mu-opioid receptor agonist, 26 nmol/microliters per h) or butorphanol (a mixed mu/delta/kappa-opioid receptor agonist, 26 nmol/microliters per h) through osmotic minipumps for 3 days. An i.c.v. injection of glutamate (5 and 50 nmol/5 microliters) dose dependently induced withdrawal signs in morphine- or butorphanol-dependent animals. The withdrawal signs precipitated by the glutamate injection were comparable to those precipitated by an opioid receptor antagonist, naloxone (48 nmol/5 microliters), except for the expression of some specific behaviors and the duration of withdrawal signs. Glutamate or naloxone challenge failed to precipitate any withdrawal signs in saline controlled animals. On the other hand, the expression of the withdrawal signs precipitated by glutamate or naloxone in opioid-dependent animals was completely blocked by pretreatment with MK-801 [a NMDA receptor antagonist, (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cycloheptan-5,10-imine], 0.1 mg/kg, i.p. These unique actions of glutamate in continuously opioid-infused rats suggest that a rapid central release of glutamate may be a key factor in the expression of withdrawal signs from opioids. Furthermore, this effect may be mediated by the NMDA subtype of glutamate receptors.

Animals↗

Differential motility stimulation but not growth stimulation or adhesion of metastatic human colorectal carcinoma cells by target organ-derived liver sinusoidal endothelial cells.

Liver is the most common distant metastatic site for colorectal cancers and when blood-borne colorectal cancer cells reach the liver, they first encounter hepatic capillary and sinusoidal endothelial cells. Thus we studied differences between highly (HT-29LMM) and poorly (HT-29P) liver-metastatic sublines of human colorectal cancer cells by examining the interactions between tumor cells and liver microvessel endothelial cells. Using hepatic sinusoidal endothelial (HSE) and lung microvessel endothelial (MLE) cell-conditioned medium we measured the growth and motility stimulating activities released from these endothelial cells and adhesion of these cancer cells to the endothelial cells. Differences in the ability of HSE-conditioned medium (HSE-CM) or MLE-conditioned medium (MLE-CM) to stimulate HT-29 cell growth were not observed. There was a small but significant increase in the rate of adhesion of highly metastatic HT-29LMM cells to HSE cell monolayers than poorly metastatic HT-29P cells, but there was no difference in adhesion to MLE cell monolayers. HSE-CM stimulated the motility of highly metastatic colorectal cancer cells to a greater extent than the poorly metastatic cells. Motility-stimulating activity for the colorectal cancer cell lines was not detected in MLE-CM. The HSE-CM motility-stimulating activity for human HT-29 cells was not removed using antibodies against hepatocyte growth factor (HGF/SF), complement component C3 or laminin, indicating that it is not related to these known liver-derived motility factors. The results suggest that the ability of highly metastatic HT-29LMM colorectal cancer cells to colonize the liver is related to their ability to respond to liver sinusoidal endothelial cell-derived motility factors and to a lesser degree to adhere to liver sinusoidal endothelial cells.

Animals↗

Increase in portal blood interleukin-6 soon after the commencement of digestive surgery.

To determine whether cytokines produced in the operative field during digestive surgery selectively spill over into the portal blood, the changes in interleukin-6 (IL-6) levels in portal and peripheral venous blood were assayed at several points in time from the commencement of surgery until 14 days later, in 11 patients. Similar changes in the IL-6 levels were observed in the portal and peripheral blood samples; however, the IL-6 levels in the portal blood reached a maximum 6-12h after the commencement of surgery, being earlier than in the peripheral venous blood. In fact, between 3 and 12h after the commencement of surgery, the IL-6 levels were higher in the portal blood by 33-81 pg/ml. By 24h or more after the commencement of surgery, the IL-6 levels did not differ significantly in the two types of blood samples. Moreover, the C-reactive protein levels 2 days after surgery were even more closely correlated to the maximum IL-6 levels in the portal blood than to those in the peripheral venous blood. These results suggest that IL-6 produced during intra-abdominal digestive surgery initially enters the portal blood, being trapped by IL-6 receptors in the liver, where it may regulate the synthesis of acute-phase proteins as a hepatocyte-stimulating factor.

Adult↗

Protective effects of antithrombin III supplementation on warm ischemia and reperfusion injury in rat liver.

The effect of antithrombin III (AT III) supplementation on energy status, microcirculation, cytoprotection, and prostacyclin (PGI2) production during and after a period of warm ischemia of the rat liver was investigated. AT III supplementation (250 units/kg) stimulate prostaglandin I2 (PGI2) production from 1 hour after administration, with maximal production observed at 3 hours. Ischemia was induced by occluding the hepatoduodenal ligament for 30 minutes, and experiments were continued for 60 minutes after reperfusion. The rats received AT III (250 units/kg IC) 30 minutes before induction of liver ischemia (AT III group). In the AT III group, recovery of the beta-ATP/inorganic phosphate ratio measured by 31P nuclear magnetic resonance showed significant improvement (p < 0.01), and the recovery of tissue blood flow markedly improved (p < 0.01) compared to the saline-treated group (control group). Leakages of aspartame aminotransferase, alanine aminotransferase, and lactate dehydrogenase were mitigated in the AT III group (p < 0. 05). Ultrastructural alterations of sinusoidal endothelial cells were markedly reduced in the AT III group. The PGI2 level at the end of reperfusion was significantly elevated (p < 0.01) in the AT III group compared to the control group. The results of this study indicated that pretreatment with AT III significantly improved the energy status and microcirculation, as well as histologic damage, after liver ischemia and reperfusion. One of the fundamental effects of AT III might be mediated through the production of prostacyclin.

6-Ketoprostaglandin F1 alpha↗

Naloxone-precipitated changes in biogenic amines and their metabolites in various brain regions of butorphanol-dependent rats.

Influence of a naloxone (an opioid receptor antagonist) challenge (5 mg/kg, IP) on levels of biogenic amines and their metabolites in various brain regions of rats infused continuously with butorphanol (a mu/delta/kappa mixed opioid receptor agonist; 26 nmol/microliter/h) or morphine (a mu-opioid receptor agonist; 26 nmol/microliter/h) was investigated using high-performance liquid chromatography with electrochemical detection (HPLC-ED). Naloxone precipitated a withdrawal syndrome and decreased the levels of: dopamine (DA) in the cortex and striatum, 3,4-dihydroxyphenylacetic acid (DOPAC) in the striatum, homovanilic acid (HVA) in the striatum, limbic, midbrain, and pons/medulla regions in butorphanol-dependent rats. However, the levels of norepinephrine (NE), serotonin (5-hydroxytryptamine; 5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) in the regions studied were not affected by naloxone-precipitated withdrawal. In addition, naloxone increased the HVA/DA ratio in the cortex, while this ratio was reduced in the limbic, midbrain, and pons/medulla. The reduction of 5-HIAA/5-HT ratio was also detected in the limbic area. In the animals rendered dependent on morphine, the results obtained were similar to those of butorphanol-dependent rats except for changes of 5-HIAA levels in some brain regions. These results suggest that an alteration of dopaminergic neuron activity following a reduction of DA and its metabolites in specific brain regions (e.g., striatum, limbic, midbrain, and pons/medulla) play an important role in the expression of the opioid withdrawal syndrome.

Animals↗

Cooperative anti-Candida effects of lactoferrin or its peptides in combination with azole antifungal agents.

The effects of lactoferrin (LF), an antimicrobial protein secreted in body fluids, and its peptides in combination with azole antifungal agents were investigated by the micro-broth-dilution method in a study of Candida albicans. In the case of LF, its pepsin hydrolysate (LFhyd) or the LF-derived antimicrobial peptide Lactoferricin B (LF-B), the concentrations required to inhibit the growth of Candida decreased in the presence of relatively low concentrations of clotrimazole (CTZ). The minimum inhibitory concentration (MIC) of all azole antifungal agents tested was reduced by 1/4-1/16 in the presence of a sub-MIC level of each of these LF-related substances. Polyene and fluoropyrimidine antifungal agents did not show such a combined effect with these LF-related substances. The anti-Candida activity of LF or LF-B in combination with CTZ was shown to be synergistic by checkerboard analysis. These results indicate that LF-related substances function cooperatively with azole antifungal agents against C. albicans.

Antifungal Agents↗

Purification and characterization of a 39kDa apurinic/apyrimidinic endonuclease from mouse ascites sarcoma cells.

We purified an apurinic/apyrimidinic (AP) endonuclease from mouse ascites sarcoma (SR-C3H/He) cells. The enzyme showed nicking activity on acid-depurinated DNA but not on untreated, intact DNA. It also showed priming activity for DNA polymerase on both acid-depurinated and bleomycin-damaged DNA. The priming activity on bleomycin-damaged DNA was two times higher than that on an acid-depurinated DNA. The enzymatic properties indicate that the enzyme is a class II AP endonuclease having DNA 3' repair diesterase activity. The purified enzyme has a molecular weight of 39,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimal pH for AP endonuclease activity was 8.0 in 50 mM Tris-HCl buffer. The AP endonuclease activity depended on divalent cation such as Mg2+ and Co2+ ions, and was inhibited by 2 mM EDTA with no addition of the divalent cation. An appropriate concentration of sodium or potassium salt stimulated the activity. Partial digestion of the AP endonuclease with Staphylococcus aureus V8 protease produced 4 major peptide fragments which may be used for protein sequencing.

Acids↗

cDNA cloning of rat major AP endonuclease (APEX nuclease) and analyses of its mRNA expression in rat tissues.

APEX nuclease is a mammalian DNA repair enzyme having apurinic/apyrimidinic (AP) endonuclease, 3'-5'-exonuclease, DNA 3' repair diesterase and DNA 3'-phosphatase activities. It is also a redox factor (Ref-1), stimulating DNA binding activity of AP-1 binding proteins such as Fos and Jun. In the present paper, a cDNA for the enzyme was isolated from a rat brain cDNA library using mouse Apex cDNA as a probe and sequenced. The rat Apex cDNA was 1221 nucleotides (nt) long, with a 951-nt coding region. The amino acid sequence of rat APEX nuclease has 98.4% identity with mouse APEX nuclease. Using the rat Apex cDNA as a probe for Northern blot analysis, the size of rat Apex mRNA was shown to be approximately 1.5 kb. Its expression was compared in 9 rat organs on postnatal days 7 and 28. Although Apex mRNA was expressed ubiquitously, the levels varied significantly, suggesting organ- or tissue-specific expression of the Apex gene. The highest level was observed in the testis, relatively high levels in the thymus, spleen, kidney and brain, and the lowest level in the liver. The level of expression at postnatal day 28, with the exception of the testis, was almost the same as or lower in respective organs than that at postnatal day 7. Postnatal developmental changes of Apex mRNA expression in the testis and thymus were further studied. The expression in testis was markedly increased on postnatal days 21 and 28. The expression in thymus increased once at postnatal day 14, and then decreased. The developmental changes of Apex mRNA expression in testis and thymus suggest that APEX nuclease is involved in processes such as recombinational events.

Amino Acid Sequence↗

Importation of aquaculture seedlings to Japan.

Japan is one of the largest consumers of fishery products in the world. Importation of various fishery commodities - including larvae and eggs for aquaculture - is increasing due to high consumer demand. Elvers of Anguilla anguilla have been imported from Europe since the late 1960s. Eyed eggs of coho salmon (Oncorhynchus kisutch) are imported from North America. During the last decade, over-production caused the price of Japanese amberjack (or 'yellowtail') (Seriola quinqueradiata) and Japanese seabream (Pagrus major) to drop, forcing farmers to venture into the culture of other species. A variety of marine fish seedlings are now imported from Asian countries. Almost all aquaculture production of coho salmon, greater amberjack (Seriola dumerili), seabass (Lateolabrax japonicus), rockfish (Sebastes spp.) and groupers (Epinephlus septemfaciatus, etc.) depends on imported seedlings. These species accounted for approximately 9% by value of marine finfish production in Japan in 1993. Apart from ubiquitous or endemic pathogens, a considerable number of exotic microorganisms and parasites are suspected to have accompanied fish eggs and larvae imported for aquaculture purposes. On a voluntary basis, 80-90% of imported salmon eggs are being examined for bacterial kidney disease and salmon viruses by semi-governmental organisation. Voluntary inspections are also being conducted on some other fish species imported from foreign countries. No legislation exists, however, for the control of exotic fish pathogens.

Animals↗

Hereditary deficiency of lactate dehydrogenase H-subunit.

We report herein the fifth family of hereditary deficiency of lactate dehydrogenase (LDH) H-subunit with an autosomal recessive inheritance including two cases of complete deficiency. Their LDH activities were low both in the serum and in the red blood cells (RBC). Electrophoretic analysis revealed that the patients with the complete deficiency had only the LDH5 isozyme. The complete deficiency was associated with marked elevation of fructose-1, 6-diphosphate (FDP) and dihydroxyacetonephosphate (DHAP) and a less marked rise in glyceraldehyde-3-phosphate (GA3P) among glycolytic intermediates in the RBC. Furthermore, hemolysis was observed in the present cases, but this finding was not included in the other reports.

Consanguinity↗