Radiotherapy is commonly used in treating many kinds of cancers which cannot be cured by other therapeutic strategies. However, radiotherapy also induces the damages on the normal tissues. Radiation-induced fibrosis is frequently observed in the patients undergoing radiotherapy, and becomes a major obstacle in the treatment of intrahepatic cancer. Hedgehog (Hh) that is an essential in the liver formation during embryogenesis is not detected in the healthy liver, but activated and modulates the repair process in damaged livers in adult. The expression of Hh increases with the degree of liver damage, regulating the proliferation of hepatic progenitors and hepatic stellate cells (HSC). In addition, Hh induces epithelial-to-mesencymal transition (EMT) and activation of myofibroblasts. In the irradiated livers, up-regulated expression of Hh signaling was associated with proliferation of progenitors, EMT induction, and increased fibrosis. Female-specific expression of Hh leaded to the expansion of progenitors and the accumulation of collagen in the irradiated livers of female mice, indicating that gender disparity in Hh expression may be related with radiation-susceptibility in female. Hence, Hh signaling becomes a novel object of studies for fibrogenesis induced by radiation. However, the absence of the established experimental animal models showing the similar physiopathology with human liver diseases and fibrosis-favorable microenvironment hamper the studies for the radiation-induced fibrosis, providing a few descriptive results. Therefore, further research on the association of Hh with radiation-induced fibrosis can identify the cell and tissue-specific effects of Hh and provides the basic knowledge for underlying mechanisms, contributing to developing therapies for preventing the radiation-induced fibrosis.
Proceedings of the Korean Society for Food Science of Animal Resources Conference
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2004.05a
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pp.89-119
/
2004
This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.
Background : Several studies have suggested that impaired mucociliary clearance plays a role in the pathophysiology of bronchial asthma. Cough productive of mucoid sputum is common, and mucous plugs in the airways are frequently observed. These clinical features are in keeping with the histologic lesions of asthma, which involve primarily the epithelial and mucous-producing structures of the conducting airways. Some studies have shown that the mucociliary clearance is impaired in adult asthma, but it has not been studied in childhood asthma. The objectives of this study were to examine whether the mucociliary clearance is impaired in childhood asthma and to estimate the degree of impairment in comparison with that of immotile cilia syndrome. Method : Thirteen children with mild stable asthma and eight patients with immotile cilia syndrome completed this study. Ten healthy children were recruited as a normal control group. The whole-lung mucociliary clearance was measured by the radioaerosol technique. Aerosols, tin colloid particles tagged with the radionuclide technetium-99m($^{99m}Tc$), were generated by means of nebulizer, and inhaled via a mouthpiece. The retention of radioactivity was measured at 30, 60, 90 and 120 minutes by gamma camera, and mucociliary clearance was calculated as percent retention at each time. Results: 1) In each subject, the percent retention decreased variably with the lapse of time. 2) The percent retention of radionuclide decreased at each time in order of normal control, bronchial asthma and immotile cilia syndrome and the percent retention of immotile cilia syndrome was significantly higher than that of normal control at each time(p<0.05). 3) At two hours, the percent retention of bronchial asthma($65.0{\pm}1.8$(SE)%) was significantly higher than that of the normal control($54.4{\pm}3.5%$, p<0.05), and significantly lower than that of immotile cilia syndrome($73.3{\pm}1.4%$, p<0.01). 4) When the percent retention was analyzed according to $PC_{20}$ in the children with bronchial asthma, they had no relationship with each other. Conclusion: Mucociliary clearance in the children with bronchial asthma was significantly lower than normal control. This finding indicates that impaired mucociliary clearance operates in childhood asthma as well, and suggests that it may be one contributing factor in the pathogenesis of asthma. The degree of impairment, however, was not so severe as immotile cilia syndrome.
This experiment was performed to evaluate the morphological responses of the gastric epithelial cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (tumor control group and BCG-treated group). In the experimental groups, each mouse was inoculated with $1{\times}10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8{\sim}0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of saline or BCG, each mouse was injected with a single dose of 0.7 ${\mu}Ci/g$ of methyl-$^3H$-thymidine (25 Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and gastric tissues were taken and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM-1 (Amersham Lab., England) in a dark room. The number of labeled epithelial cells in the gastric mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. And for electron microscopic observation, gastric tissues were prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. On the light microscopic study, gastric mucosae had no morphological changes following the injection of BCG. On the electron microscopic study, in the BCG-treated mice, myelin figures and multivesicular bodies within the gastric epithelial cells were observed more frequently than in those of the normal control ones. On the autoradiographic study, number of the labeled cells of normal control, tumor control and BCG-treated mice were 380.2 (${\pm}31.35$), 426.1 (${\pm}28.43$) and 301.8 (${\pm}34.63$), respectively. In the BCG-treated mice, poorly-labeled cells containing only a few silver grains of 3H-thymidine were observed more frequently as compared in those of the normal control and tumor control ones. From the above results, BCG may suppress the DNA synthesis of the gastric epithelial cells, but does not results severe fine structural defect on the gastric epithelial cells. These results suggest that BCG is expected as one of the effective supplemental anticancer drugs.
The effect of intravesical formalin instillation as a therapeutic modality for intractable bladder hemorrhage is well known. And despite clear evidence of therapeutic efficacy of intravesical cytotoxic drugs and/or BCG immunotherapy, there have been substantial recurrences during followup after transurethral resection for superficial bladder tumor. If formalin injected at the bed of superficial bladder tumor is able to coagulate and necrotize the tumor, it will be greatly helpful to the patients With recurrent bladder tumor developed during followup. Since this technique is applicable on outpatient basis, an economical as well as a psychological burden of the patients can be reduced considerably. The purpose of this study is to evaluate the effect of submucosal formalin injection on rat bladder wall, 36 healthy adult male Sprague-Dawley rats (weighing 350gm in average) were divided into 3 groups: In Group I (control group), 0.01ml of normal saline was injected submucosally at the left posterolateral wall of the bladder opened under intraperitoneal Nembutal anesthesia ; In Group II and III, 0.01 ml of 10% and 4% formalin, respectively, were administered at the same site as in the Group I, two rats in each group were sacrificed at day 1, 2, and 3, and week 1, 2 and 4 after injection, respectively. Gross and microscopic examination of the cystectomized specimen were done in each group. In the Group II, bladder stones were formed at week I, and in both the Group I and III, stones were seen at week 2 post injection. There was no significant difference III histologic findings of the bladder between the group II and III. Mucosal ulcer and/or prominent mucosal disruption was observed at 24 hours after injection in both Group II and III. Epithelial regeneration began at day 2, and was marked at day 3, and epithelial lining was almost normalized one week after injection. Subepithelial edema, telangiectasia and inflammatory reaction were prominent at 24 hours post formalin injection. Subepithelial edema persisted in moderate degree for 1 week. Telangiectasia and inflammatory reaction were noted for 4 weeks. Mild degree of these findings also appeared In the control group. Fibroblastic proliferation appeared at day 2 and persisted in moderate degree for 4 weeks. There has been no mortality or bladder perforation. These results suggest that clinical application of this technique is feasible for the selected cases of recurrent, solitary superficial bladder tumor. However, optimal dosage of formalin in relation to the size of the lesion remains to be investigated.
This study was performed to determine the minimum dose of Poloxamer/Sodium alginate (PX/SA) mixture on preventing intraperitoneal adhesions to evaluate organ toxicity. Twenty five healthy adult mongrel dogs (weighing 4.68${\pm}$1.67 kg) were divided into five experimental groups composed of five dogs respectively; negative control group (NC, non-treated), positive control group (PC, 2% carboxymethyl chitosan solution treated), and experiment 1 group (E1, 0.25 ml PX/SA mixture of abraded area), experiment 2 group (E2, 0.5 ml PX/SA mixture of abraded area), experiment 3 group (E3, 1.0 ml PX/SA mixture of abraded area). Venous blood specimens were collected from all experimental animals for hematologic and biochemical analysis: WBC, fibrinogen, AST, ALT, ALP, BUN and creatinine. The anti-adhesion effect was evaluated using a serosa abrasion model. The denuded ileum was coated with PX/SA mixture, carboxymethyl chitosan solution or neither. The tensile strength of the adhesion site was evaluated with a tensiometer. For histopathological examination, tissue samples of the liver and kidney were collected from all dogs. According to the results, the frequency and tensile strength values for adhesion separation in PX/SA group were significantly lower than those in negative control group (p < 0.05). In E2 group, the tensile strength was significantly decreased in consideration of PX/SA dose. The values of AST, ALT, ALP, BUN and creatinine of the control and the experimental groups showed no statistical differences. No obvious microscopic differences were noted among tissue sections obtained from all groups. The results suggest that PX/SA mixture may be effective on reducing peritoneal adhesion formation in dog and that 0.5 ml PX/SA mixture of abraded area is most effective dose. Moreover, PX/SA mixture was considered not to have toxicity for the liver and the kidney.
Non-alcoholic fatty liver disease accompanies the rise in the prevalence of obesity, diabetes and the tendency toward high-fat dietary habits. Specifically, the higher prevalence of non-alcoholic fatty liver disease in men and postmenopausal women seems to be caused by the protective effects of estrogen against liver fibrosis, or lack thereof. There are no effective preventive therapies for liver diseases because the mechanisms underlying the progression of fatty liver diseases to chronic liver diseases and the protective effects of estrogen against fibrogenesis remain unclear. Recently, it has been reported that the hedgehog signaling pathway plays an important role in the progression of chronic liver diseases. Hedgehog, a morphogen regulating embryonic liver development, is expressed in injured livers but not in adult healthy livers. The level of hedgehog expression parallels the stages of liver diseases. Hedgehog induces myofibroblast activation and hepatic progenitor cell proliferation and leads to excessive liver fibrosis, whereas estrogen inhibits the activation of hepatic stellate cells to myofibroblasts and prevents liver fibrosis. Although the mechanism underlying the opposing actions of hedgehog and estrogen on liver fibrosis remain unclear, the suppressive effects of estrogen on the expression of osteopontin, a profibrogenic extracellular matrix protein and cytokine, and the inductive effects of hedgehog on osteopontin transcription suggest that estrogen and hedgehog are associated with liver fibrosis regulation. Therefore, further research on the estrogen-mediated regulatory mechanisms underlying the hedgehog-signaling pathway can identify the mechanism underlying liver fibrogenesis and contribute to developing therapies for preventing the progression of fibrosis to chronic liver diseases.
Park, In Won;Koh, Hyung Ki;Kang, Yoon Jung;Choi, Jae Sun;Yoo, Jee Hoon;Shin, Jong Wook;Lim, Seong-Yong;Choi, Byoung Whui;Seo, Seung Cheon;Na, Moon Jun;Hue, Sung Ho
Tuberculosis and Respiratory Diseases
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v.44
no.1
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pp.154-161
/
1997
Background : Although the long term adverse effects of cigarette smoking on health are well known, the acute possible detrimental effects of smoking on pulmonary or cardiovascular function, especially when these systems are stressed by the metabolic demands of exercise, have not been well studied The purpose of this study is to determine the acute action of cigarette smoking on cardiopulmonary function under stress. Method : Twenty -one healthy smoking subjects were studied. Before exrecise testing, history taking, physical examination and baseline studies, including CBC, chest PA, PFT and EKG, were done. The subjects performed an incremental bicycle exercise test to exhaustion on two occasions, one without smoking and the other after smoking 5 cigarettes/h for 2 hours. All indices of P.F.T. and bicycle ergometry were compared between before and after smoking. Results : 1. $VO_2$max and $O_2$ pulse showed significant decrease in smoking day. 2. Although there were no significant differences, anaerobic threshold showed a tendency of decrease and HRmax showed that of increase in smoking day. 3. P.F.T. and respiratory indices showed no significant change io smoking day. Conclusion : Cigarette smoking has immediate adverse effect, especially on the cardiovascular system rather than the respiratory system. These results would be due to the effect of elevated HbCO and/or impaired blood flow in response to the exercise stimulus.
Background : In acute lung injury, activated neutrophils play an important role in tissue damage. For neutrophils to participate in lung inflammation, chemotactic factors released from mononuclear phagocytes are needed to bring these cells to the local site of inflammation, with interleukin-8 (IL-8) being one of the most specific and important chemotactic factors for neutrophils. IL-8 also induces the expression of adhesion molecules and activates neutrophils to release various inflammatory mediators. Lipopolysaccharide(LPS) is one of the most important causes of adult respiratory distress syndrome and can cause release of many inflammatory cytokines including IL-8 leading to acute lung injury. But little is known about the regulatory mechanism of LPS-induced IL-8 gene expression in mononuclear phagocytes. Method : Human alveolar macrophages(HAM) and peripleral blood monocytes(PBMC) were isolated from healthy volunteers. Time and dose relationship of LPS-induced IL-8 mRNA expression was observed by Northern blot analysis. To evaluate the regulatory mechanism of LPS-induced IL-8 gene expression, pretreatment of actinomycin D(AD, $5{\mu}g/ml$) and cycloheximide(CHX, $5{\mu}g/ml$) was done and Northern blot analysis for IL-8 mRNA and ELISA for immunoreactive IL-8 protein in culture supernatant were performed. Results : 1) In HAM, dose and time dependent LPS-induced IL-8 mRNA expression was observed with peak mRNA level at 8 hours post-stimulation. 2) In PBMC, dose and time dependent LPS-induced IL-8 mRNA expression was also observed with peak mRNA level at 4 hours post-stimulation. 3) AD decreased expression of LPS-induced IL-8 gene expression at both mRNAand protein levels in both types of cells. 4) CHX decreased expression of LPS-induced IL-8 gene expression at protein level in both cell types but in HAM, superinduction of IL-8 mRNA was observed while decreased expression of IL-8 mRNA was observed in PBMC. Conclusion : Time and dose dependent LPS-induced IL-8 gene expression was observed in mononuclear phagocytes which is at least partly regulated pretranslationally. LPS-induced IL-8 mRNA expression in HAM needs no de novo protein synthesis and may be under the control of a labile repressor protein while de novo protein synthesis may be needed in PBMC.
In order to evaluate the significance of thyrotropin-binding inhibiting immunoglobulin (TBII) in the patients with autoimmune thyroid diseases, the authors investigated 402 cases of Graves' disease and 230 cases of Hashimoto's thyroiditis comparing 30 cases of normal healthy adult at Kyung Pook University Hospital from February 1993 to August 1994. The TBII was tested by radioimmunoassay and assesed on the dynamic change with the disease course, thyroid functional parameters, and other thyroid autoantibodies : antithyroglobulin antibody (ATAb) and antimicrosomal antibody (AMAb) including thyroglobulin. The serum level of TBII was $40.82{\pm}21.651(mean{\pm}SD)%$ in hyperthyroid Graves' disease and $8.89{\pm}14.522%$ in Hashimoto's thyroiditis and both were significant different from normal control of which was $3.21{\pm}2.571%$. The frequency of abnormally increased TBII level was 92.2% in hyperthyroid Graves' disease, 46.7% in euthyroid Graves' disease or remission state of hyperthyroidism, and 23.9% in Hashimoto's thyroiditis. The serum levels of increased TBII in Graves' disease were positively correlated with RAIU, serum T3, T4, and FT4, but negatively correlated with serum TSH(each P<0.001). The TBII in Graves' disease had significant positive correlation with serum thyroglobulin and AMAb, but no significant correlation with ATAb. In the Hashimoto's thyroiditis, the serum levels of TBII were positively correlated with RAIU, serum T3, TSH and AMAb, but not significantly correlated with serum T4, FT4, thyroglobulin and ATAb. Therefore serum level of TBII seemed to be a useful mean of assessing the degree of hyperthyroidism in Graves' disease and correlated well with thyroidal stimulation. The serum level of TBII in Hashimoto's thyroiditis is meaningful for the degree of both functional abnormality reflecting either hyperfunction or hypofunction and the immune logic abnormality.
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