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Biomedical subjects

K Honda

Publications and source records attributed to K Honda.

At least 181 records · Page 10Linked to original sources

Preservation of pathological tissue specimens by freeze-drying for immunohistochemical staining and various molecular biological analyses.

Conditions of preserving DNA, RNA and protein in pathological specimens are of great importance as degradation of such macromolecules would critically affect results of molecular biological analysis. The feasibility of freeze-drying as a means of preserving pathological tissue samples for molecular analysis has previously been shown. In the present study, further tests on long-term storage conditions and analyses of freeze-dried samples by polymerase chain reaction (PCR), reverse transcriptase (RT)-PCR, western blotting and immunohistochemistry are reported. Rat chromosomal DNA of freeze-dried samples stored for 4 years showed slight degradation while RNA degradation was more prominently seen at an earlier stage of storage. However, these 4 year DNA and RNA samples were still able to serve as a template for some PCR and RT-PCR analyses, respectively. Overexpression of c-erbB-2 and p53 protein was demonstrated by western blotting and immunohistochemical staining using freeze-dried human breast cancer tissues. Although macromolecules in freeze-dried samples degrade to some extent during the preservation period, they should still be of value for certain molecular biological analyses and morphological examination; hence, providing more convenient and inexpensive ways of pathological tissue storage.

Animals↗

Protective effect of lactosucrose on intracolonic indomethacin-induced small-intestinal ulcers in rats.

BACKGROUND: Little is known about the role of intestinal microflora in the development of indomethacin-induced enteropathy. The aim of this study was to evaluate the effects of lactosucrose, an indigestible oligosaccharide, on intestinal microflora and on indomethacin-induced enteropathy in rats. METHODS: Male Wistar rats were fed either sucrose (SC) or lactosucrose (LS) for 2 weeks. Indomethacin (24 mg/kg/ day) was administered into the colon twice, 24 h apart, and intestinal ulcers in SC and LS groups were compared macroscopically. In another experiment the bacterial composition in the mid-small-intestinal segment was determined in both groups before and after treatment with indomethacin. RESULTS: After indomethacin treatment small-intestinal ulcers were less severe in the LS than in the SC group (ulcer index: median, 0.13 (range, 0.05-0.19) versus 0.23 (0.13-0.34); P < 0.05). Total bacterial count did not differ significantly between the two groups. Indomethacin increased the number of Enterobacteriaceae in both groups, but the increase was less in the LS group. The number of streptococci was also significantly increased in the SC group but not in the LS group. CONCLUSION: These results suggest that LS has some protective effects on indomethacin-induced enteropathy and that this protective effect is in part due to the maintenance of intestinal microflora.

Animals↗

Studies on the HLA-DRB1 genotypes in Japanese women with severe pre-eclampsia positive and negative for anticardiolipin antibody using a polymerase chain reaction-restriction fragment length polymorphism method.

The human leukocyte antigen (HLA)-DR genotype was determined in 54 Japanese women with severe pre-eclampsia in order to elucidate the relationship between HLA-DR antigen systems and pre-eclampsia. The patients were divided into two groups according to positivity for the anticardiolipin antibody (ACA), i.e. one patient group negative for ACA (n = 41) and the other patient group positive for ACA (n = 13). The frequency of each HLA-DRB1 allele in both groups was compared with that in 81 normally fertile Japanese women who had not experienced pre-eclampsia. The genotypes of HLA-DR antigens were determined using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The frequency of DRB1*04 and DRB1*0403 in the patient group positive for the ACA was significantly higher compared with that in the group of normal fertile women (P< 0.05). The frequency of each HLA-DRB1 allele was not significantly different between patient group with pre-eclampsia negative for ACA and group of normal fertile women. These results suggest a difference in the immunogenetic background between the patient groups with severe pre-eclampsia positive and negative for the ACA.

Adult↗

IL-7 receptor alpha+ CD3(-) cells in the embryonic intestine induces the organizing center of Peyer's patches.

Peyer's patch (PP) organogenesis proceeds through three histologically distinct steps: formation of organizing centers expressing VCAM-1 and ICAM-1 in segregated regions of the intestine at 15.5 days post-coitus (d.p.c.) (step I), accumulation of blood cells expressing different sets of surface markers to this region at 16.5-17.0 d.p.c. (step II), and entry of CD3+ and B220+ lymphocytes just before birth (step III). PP formation of both Il7ra-/- and Lta-/- mice is impaired from step I, suggesting involvement of the two molecules at the same timing in PP organogenesis. Expression of lymphotoxin (LT) alpha and LTbeta in IL-7 receptor (IL-7R) alpha+ cells in the intestine indicates that defects of Il7ra-/- and Lta-/- mice are due to functional inability of IL-7Ralpha+ cells in the induction of PP anlage. Blocking of IL-7Ralpha function by a single injection of the antagonistic mAb in 15.5 d.p.c. embryos suppressed appearance of VCAM-1(+) spots and expression of LTalpha and LTbeta in the intestine, which eventually resulted in mice without PP but are otherwise normal. Intestinal IL-7Ralpha+ cells are lymphoid in morphology but CD3(-) and functional in both nu/nu and Rag2-/- mice. These results implicate IL-7Ralpha+ CD3(-) cells as the direct inducer of the organizing center of PP.

Animals↗

Accumulation of advanced glycation end products in the peritoneal vasculature of continuous ambulatory peritoneal dialysis patients with low ultra-filtration.

BACKGROUND: Ultra-filtration failure is a serious complication of long-term continuous ambulatory peritoneal dialysis (CAPD). This complication is related to histological changes of the peritoneum, i.e. severe interstitial fibrosis and microvascular sclerosis. Although their pathogenesis has not been elucidated yet, advanced glycation end products (AGEs) have been shown to accumulate in the peritoneal tissue of CAPD patients. METHODS: Peritoneal biopsy specimens from 14 CAPD patients with low ultra-filtration (n = 9) and high ultra-filtration (n = 5) capacity were immunohistochemically investigated using a monoclonal antibody against AGEs (6D12). The severity of peritoneal fibrosis, microvascular sclerosis and intensity of AGE accumulation were semi-quantitatively evaluated. Peritoneal ultra-filtration capacity was evaluated by calculating daily ultrafiltration volume per body weight (UFV/BW) and D/D0 (glucose) of the peritoneal equilibration test. RESULTS: In all patients with low ultra-filtration, AGE accumulated in the peritoneal fibrous tissue and microvascular walls. Remarkably, AGE accumulated more intensely in hyalinized fibrosis of small venular media. Extent of AGE accumulation in peritoneal interstitium and vascular walls correlated with the progression of interstitial fibrosis (rho = 0.727, P = 0.0088) and vascular sclerosis (rho = 0.915, P = 0.001). UFV/BW was inversely correlated to interstitial fibrosis (rho = -0.660, P = 0.0174), microvascular sclerosis (rho = -0.671, P = 0.0155) and microvascular AGE accumulation (rho = -0.678, P = 0.0145). CONCLUSIONS: In CAPD patients, AGE formation in the peritoneum correlates with the development of severe interstitial fibrosis and microvascular sclerosis, which is associated clinically with impaired peritoneal ultra-filtration.

Adult↗

The hsp operons are repressed by the hrc37 of the hsp70 operon in Staphylococcus aureus.

The heat-shock proteins are coded for the polycistronic operons hsp70 and hsp60 in Staphylococcus aureus. The hsp70 operon is comprised of five genes, hrc37, hsp20, hsp70, hsp40 and orf35, and the hsp60 is comprised of two genes, hsp10 and hsp60. The hsp70 operon transcribed five different sizes of mRNA from three promoters: P1, the most active promoter, transcribed 6.0 and 3.6 kb mRNAs; P2 transcribed a single 1.8 kb mRNA; and P3 transcribed 4.2 and 2.4 kb mRNAs. The hsp60 operon transcribed a single 2.1 kb transcript from only one promoter, P1. Both operons had a common structure of inverted repeat element (CIRCE, Controlling Inverted Repeat of Chaperon Expression) at the promoter region. All of the transcripts were heat (46 C) inducible. One of the unidentified genes, hrc37, was characterized. The disruptant of the hrc37 in the hsp70 operon enhanced the transcription of both operons at 37 C (derepression). Complementation of the disruptant with the cloned hrc37 plasmid recovered the repression of the transcription of both operons at 37 C. The product of hrc37, Hrc37, was found to bind to the CIRCE element. These findings indicated that Hrc37 from the hsp70 operon repressed the transcription of both the hsp70 and hsp60 operons by binding to the CIRCE element located at the promoter region.

Base Sequence↗

Metabolic and morphologic characteristics of adipose tissue associated with the growth of malignant tumors.

Changes in total body fat and the metabolic and morphologic characteristics of adipose tissue were sequentially investigated in individual rabbits implanted with VX2 tumors to elucidate the pathology of the fat reduction in animals with malignant tumors as compared with that of diet-restricted rabbits. Lipogenesis in normal, VX2-implanted, and diet-restricted rabbit groups on day 40 after the start of the experiments was 19.1 +/- 2.9, 13.3 +/- 3.5, and 41.7 +/- 6.0 x 10(5) cpm/g/h, respectively, and glycerol liberation by their adipose tissue was 199 +/- 21, 528 +/- 94, and 301 +/- 45 nmol/g/h, respectively. In addition, apoptotic cells were noted in the adipose tissue of VX2-implanted rabbits on days 20-30 after implantation, but not in diet-restricted rabbits. The results showed clear differences between the total body fat reduction profiles of VX2-implanted rabbits and diet-restricted rabbits, suggesting a characteristic lipid metabolism with enhanced lipolysis and diminished lipogenesis in VX2-implanted rabbits. The results strongly suggest that adipocyte apoptosis might be involved in these phenomena.

Adipose Tissue↗

Hepatorenal interaction in glutathione redox state during partial hepatic ischemia-reperfusion in rats.

The hepatic glutathione metabolism during partial hepatic ischemia-reperfusion was studied in a rat model with special reference to the hepatorenal glutathione metabolism system. Bile samples, and in- and outflow blood and tissue samples of the rat liver, kidney and small intestine were collected during 60 min of 70% partial liver ischemia and reperfusion. Both reduced and oxidized glutathione levels were determined by HPLC. The tissue ratio of reduced to oxidized glutathione (GSH/GSSG ratio) decreased significantly in the ischemic hepatic lobe at 60 min after reperfusion. The GSH/GSSG ratio in bile from the ischemic hepatic lobes decreased significantly after 60 min of ischemia and gradually recovered after reperfusion. The net release of GSH from the nonischemic hepatic lobe increased at 60 min after reperfusion, since the calculated net release of GSH from the whole liver increased significantly whereas there was no change in the net release from the ischemic hepatic lobe. The tissue GSH level in the kidney increased significantly at 180 min after reperfusion. The calculated net uptake of GSH into the kidney, and the net release of total cysteine from the kidney, tended to decrease at the end of 60 min of ischemia, to increase at 60 min after reperfusion and then decrease at 180 min after reperfusion. We found that the hepatorenal glutathione metabolism was changed by partial hepatic ischemia-reperfusion. These changes might reflect a hepatorenal interaction to maintain the glutathione redox state of the vital organs.

Animals↗

Intermittent cisplatin therapy for stage-III ovarian cancer patients following clinical remission.

Intermittent cisplatin (CDDP) administration as consolidation therapy for stage-III ovarian cancer was studied. No patients with residual disease at surgery or at second-look operation or patients with <8 U/ml of CA125 after remission induction chemotherapy were enrolled in this study. Fifteen subjects received intermittent chemotherapy, and 10 control patients did not. CDDP of 20 mg/m2 for 5 days was administered every 3-4 months for 5 years. A significant increase in 5-year survival rate was found in the intermittent chemotherapy group when compared with the control group. Side effects from the intermittent chemotherapy were controllable. Intermittent CDDP therapy seems to be an effective consolidation therapy for stage-III ovarian cancer patients.

Antineoplastic Agents↗

Leiomyomas in uterine remnants associated with vaginal agenesis.

A premenopausal woman with vaginal agenesis underwent surgery for leiomyomas in rudimentary uterine horns. Immunohistochemical study of surgical specimens revealed that progesterone receptors are distributed in the nuclei of a majority of the leiomyoma cells and in the nuclei of a minority of the myometrial cells. Estrogen receptors are not detected in the surgical specimens.

Cell Nucleus↗

Eating problems and related weight control behaviour in adult Japanese women.

BACKGROUND: Fewer studies concerning eating problems have been conducted in adult than in adolescent female populations. The aims of this study are to ascertain the proportion of eating problems and clarify weight control behaviour in adult Japanese women. METHODS: This study employed a questionnaire survey with a cross-sectional design. Subjects were adult females aged 20-39 years, working in a computer factory. Four hundred and six women agreed to participate in the study. The questionnaire elicited self-reported age, body height, body weight and various histories of dieting, fasting and purging behaviour. The Eating Attitudes Test 26-item version (EAT-26) was also administered. Subjects showing an EAT-26 score equal to or greater than 20 were identified as having eating problems. RESULTS: Among the subjects, 172 women (42.4%) had some history of dieting behaviour and 24 (5.9%) had some history of fasting behaviour. Regarding misuse of laxatives, diet pills and diuretics for the purpose of weight loss, 50 (14.3%) were past or current users of diet pills, followed by 42 (10.3%) laxative abusers and 15 (3.7%) diuretic abusers. Six (1.5%) subjects had an EAT-26 score of 20 or greater, 5 (1.9%) in their 20s, and 1 (0.8%) in her 30s, the difference not being statistically significant. CONCLUSIONS: Although dieting behaviour is common in adult Japanese working women, the proportion of eating problems is low in comparison with Western adult populations.

Adult↗

Whole-blood flow-cytometric analysis of induction of adhesion molecules expression on eosinophils stimulated by phorbol-12-myristate-13-acetate/ionomycin.

BACKGROUND: Activation of eosinophils is closely associated with the pathology of allergic inflammatory disease, especially bronchial asthma. We recently investigated the activation of eosinophils by applying whole blood to a flow cytometer. We measured here beta1 and beta2 integrin on eosinophils stimulated by phorbol-12-myristate-13-acetate (PMA)/ionomycin to evaluate eosinophil activation in vitro using whole blood. METHODS: Heparinized whole blood was diluted with the same volume of RPMI 1640, then cells were incubated in the presence or absence of PMA and ionomycin for 45 min at 37 degrees C. After hemolyzation with lysing solution, flow-cytometric findings for CR3, LFA1-alpha, LFA1-beta and VLA-4 expression on eosinophils were examined. RESULTS: Mean fluorescent intensity (MFI) of CR3 and LFA1-beta stimulated by PMA and ionomycin was significantly higher than that of the unstimulated control. MFI of LFA1-alpha showed no significant difference from the unstimulated control. On the other hand, MFI of VLA-4 tended to decrease. CONCLUSIONS: Our method to distinguish eosinophils from various cell groups in whole blood is simple and time-saving, similar to conditions in vivo and may allow intensive investigation of eosinophils in clinical laboratories as well as in research laboratories. We are currently investigating the influence of different kinds of stimulations, regulation factors or agents on eosinophils using this method.

Carcinogens↗

CCR3 mRNA expression in bronchial epithelial cells and various cells in allergic inflammation.

BACKGROUND: RANTES and eotaxin are important chemokines involved in the activation and migration of eosinophils and are considered to play a major role in allergic inflammation. METHODS: In this study, we used RT-PCR to investigate the kinds of cells that express mRNA for CCR3, a common receptor of these chemokines, and eotaxin, a ligand for CCR3. RESULTS: CCR3 mRNA was expressed in eosinophils, peripheral mononuclear cells, an eosinophilic cell line (EoL-1), a bronchial epithelial cell line (NCI-H(292)), human endothelial cells and nasal washings from patients with allergic rhinitis. CONCLUSION: These results suggest that the CCR3-eotaxin system plays an important role in generating inflammation, since these substances are expressed not only in cells implicated in activation or migration of eosinophils but also in various other cells involved in allergic inflammation.

Bronchi↗

Effect of eotaxin on the generationof reactive oxygen species from eosinophil cell line, YY-1.

BACKGROUND: The CC chemokine eotaxin is a selective chemoattractant for eosinophils. Eosinophils have been considered to be the major effector cells in allergic inflammation, since not only eosinophil-specific granule proteins but also reactive oxygen species (ROS) from eosinophils may cause the damage to the cells or tissue of the mucosal epithelium. In this study, we examined the effect of eotaxin on ROS from an eosinophil cell line, YY-1. METHODS: ROS in luminol-dependent reaction were examined. Calcium ionophore A23187 were added to the mixture of YY-1 cells with luminol, and then ROS were determined. RESULTS: Eotaxin primed the production of ROS from YY-1 cells. ROS from untreated YY-1 cells evoked with calcium ionophore A23187 in luminol-dependent chemiluminescence gave a maximal value of 1,928 +/- 223 intensity counts (IC; mean +/- SE, n = 4) and an integral value of 17.04 +/- 1. 51 IC (x10(-4)), while eosinophils that were treated with eotaxin gave a maximal value of 2,264 +/- 86 IC (10 nM), 2,691 +/- 124 IC (100 nM) and an integral value of 21.22 +/- 0.67 IC (x10(-4); 10 nM), 26.20 +/- 1.41 IC (x10(-4); 100 nM). CONCLUSION: Eotaxin might play important roles in the pathogenesis of allergic inflammation through eosinophil activation by priming of eosinophil oxidative metabolism as well as involvement in selective eosinophil chemotaxis.

Cell Line↗

Growth hormone-releasing hormone and morphine attenuate growth hormone secretagogue-induced activation of the arcuate nucleus in the male rat.

Growth hormone secretagogues (GHS) administered systemically selectively induce growth hormone (GH) release from the pituitary and the expression of Fos protein in arcuate nucleus neurons. Both the control of GH release and the expression of the GHS receptor in the arcuate nucleus are thought to be regulated, at least in part, by the negative feedback actions of GH. In this study, we utilized the immunocytochemical detection of Fos protein to examine the effects of morphine- and GH-releasing hormone (GHRH)-induced GH release on the activation of arcuate nucleus neurons following GHS administration. Given alone, two structurally different GHS induced significant amounts of Fos-LI in the arcuate nucleus of male rats, suggesting activation of cells in this region. Prior administration of morphine or GHRH significantly reduced the number of Fos-positive cells in the arcuate nucleus of rats injected with either GHS, although when given together, morphine and GHRH did not produce a greater reduction in Fos expression than when given alone. In no case was there a complete reduction in Fos expression, indicating that some arcuate nucleus neurons are not subject to the feedback effects of endogenous GH. These results provide evidence that, in the male rat, GH can feedback to the hypothalamus, altering the responsiveness of neurons involved in the central response to GHS.

Animals↗

Inhaled nitric oxide reduces tyrosine nitration after lipopolysaccharide instillation into lungs of rats.

Nitric oxide (NO) may either protect against or contribute to inflammatory lung injury. In this study we investigated whether inhalation of 20 ppm NO alters tyrosine nitration, and we assessed the degree of lung inflammation and edema in rats after lipopolysaccharide (LPS) instillation. The amount of nitrotyrosine relative to the total amount of tyrosine was measured in lung homogenates, and lung tissue sections were stained for nitrotyrosine and aminotyrosine (a reduced form of nitrotyrosine). Leukocytes in bronchoalveolar lavage fluid (BALF) were counted, and myeloperoxidase activity was measured in lung homogenate. Lung edema and inflammatory cell accumulation in lung tissue were estimated by extravascular lung water weight (EVLW) and extravascular dry lung weight (EVDW), respectively. LPS instillation caused increases in nitrotyrosine concentration and immunohistochemical staining of nitrotyrosine and aminotyrosine in the lungs. LPS instillation increased the BALF leukocyte count, myeloperoxidase activity in lung tissue, and both EVLW and EVDW. Inhalational exposure to 20 ppm NO induced nitrotyrosine and aminotyrosine formation only in bronchial epithelial cell surface of the lungs not instilled with LPS. NO inhalation reduced the increases in nitrotyrosine and aminotyrosine in LPS-instilled lung tissue as well as the leukocyte count in BALF and myeloperoxidase activity in lung tissue, but it did not significantly change EVLW or EVDW. Leukocyte depletion in LPS-instilled rats reduced interstitial inflammatory cells, which were stained with nitrotyrosine and aminotyrosine, and attenuated the nitrotyrosine staining of alveolar capillaries. These results suggest that inhalation of 20 ppm NO reduces leukocyte accumulation in the lungs and inhibits tyrosine nitration caused by LPS instillation.

Administration, Inhalation↗

Lipolytic activity of anemia-inducing substance from tumor-bearing rabbits.

Anemia-inducing substance (AIS) is a protein of approximately 50,000 molecular weight secreted by malignant tumor tissue that depresses erythrocyte and immuno-competent cell functions; in this study, its biological effects on adipocytes were examined. Changes in body weight, total body fat, and food intake were investigated in rabbits after VX2 carcinoma transplantation, and the results showed reductions of 11%, 24%, and 30%, respectively, at 40 days after transplantation compared with baseline values (before transplantation). The values were even more markedly reduced 70 days after transplantation. When cyclic plasma perfusion (2 times/wk) was started at 40 days after transplantation, the values at 70 days after transplantation (30 days after beginning plasma perfusion) recovered to 91%, 84%, and 87%, respectively, of the baseline values. AIS fractions were isolated from rabbit plasma by using a phenyl-Sepharose column before transplantation, 40 and 70 days after transplantation, and 30 days after start of plasma perfusion, and AIS activity and lipolytic activity were measured. The results showed enhancement of AIS activity and lipolytic activity as the tumors grew. Lipolytic activity also returned to baseline value as AIS was removed by adsorption by plasma perfusion, and there was a high correlation between lipolytic activity and AIS kinetics. These results strongly suggest that AIS might be one of the substances involved in the enhanced lipolytic activity in advanced tumor-bearing subjects.

Adipose Tissue↗