Objectives : Iron (Fe) is an essential element in biological processes; however excessive Fe is harmful to human health. Some air pollutants contain a high level of Fe, and the human lung could therefore be over-exposed to Fe through inhaled air pollutants. This study was performed to investigate the role of metal transporters (divalent metal transporter 1, DMT1, and metal transporter protein 1, MTP1) in the lung under the environments of Fe deficiency in the body and Fe over-exposure in the lung. Methods : Rats were fed Fe deficient (FeD, 2-6 mg Fe/kg) or Fe supplemented (FeS, 120 mg Fe/kg) diet for 4 weeks, followed by a single intratracheal instillation of ferrous sulfate at low (10 mg/kg) or high (20 mg/kg) dose. Fe concentration was analyzed in the serum, lung and liver, and histopathological findings were observed in the lung at 24 hours after Fe administration. The level of DMT1 and MTP1 expression in the lung was analyzed by RT-PCR. Also, the effect of Fe deficiency in the body was evaluated on the level of Fe concentration and metal transporters compared to FeS-diet fed rats at the end of 4-week FeD or FeS diet. Results : The 4-week FeD diet in rats induced an Fe deficiency anemia with decreased serum total Fe, increased unsaturated Fe binding capacity and hypochromic microcytic red blood cells. The concentration of Fe in the lung and liver was lower in the FeD-diet fed rats than in the FeS-diet fed rats. The level of metal transporters mRNA expression was higher in the FeD-diet fed rats than in the FeS-diet. The concentration of Fe in the lung was increased in a dose-dependent pattern after intratracheal instillation of Fe into the rats, while the level of Fe in the serum and liver was not increased in the low-dose Fe administered rats. Therefore, DMT1 and MTP1 mRNA was highly expressed in both FeD-diet and FeS-diet fed rats, after intratracheal instillation of Fe. Conclusions : DMT1 and MTP1 mRNA were more highly expressed in FeD-diet fed rats than in FeS-diet fed rats. The over-exposure of Fe intratracheally induced high expression of metal transporters and increased Fe deposition in the lung in both FeD-diet and FeS-diet fed rats, but did not increase the Fe level of the serum and liver in low-dose Fe administered rats. These results suggest that the role of metal transporters in the lung might be different in a part from the duodenum under the environment of over-exposure to Fe.
Three experiments were conducted to investigate the effect of feeding yeast accumulated transgenic ferritin(FRT, Saccharomyces cerevisiae) as a probiotic on the performance, iron contents in the liver, spleen, bone and yolk of laying hens and broiler chicks. Effects of feeding FRT were compared with that of feeding wild-type yeast(W0) and yeast grown on 20 mM ferric citrate-added medium (W20). In Expt 1, to investigate the effect of feeding yeast (control, W0 FRT) on performance and iron content of organs of broiler chicks which were fed basal diet supplemented with 75mg/kg iron(Fe75) or not (Fe0), three hundred sixty one-day-old male broiler chicks were fed a corn-sov based diet for five weeks. Weight gain, feed intake and feed conversion were measured weekly. In Expt 2, fifteen 33-week-old ISA Brown laying hens were placed in individual cages and were fed control, W0 and FRT diets for Four weeks. In Expt 3, twenty four 45-week-old ISA Brown laying hens were placed in individual cages and were fed a basal diet for a week. Then, experimental diets (control, W0, W20, FRT) were fed for three weeks. Iron contents in the liver, heart, spleen and tibia were determined at the end of all experiments. Iron content in yolk was measured weekly (expt 2, 3). The level of yeast added and iron concentration of FRT were $1{\times}10^8$cfu/kg diet and 500 mg/kg cell (DM) respectively in Expt 3, yeast was supplemented at $2{\times}10^{10}$cfu/kg diet and the iron content of FRT was 1000mg/kg cell (DM). In Expt 1. birds fed Fe75 showed significantly higher weight gain compared with Fe0 (P<0.05). However, weight gain and feed intake of birds fed FRT was significantly lower than control (P<0.05). In Expt 2, the iron content of the liver was decreased in the FRT treatment (P<0.05). In Expt 3, iron concentration of the liver and spleen tended to be increased by feeding FRt. However, the iron content of the tibia tended to be decreased in the FRT treatment. These results suggest that feeding FRT as a probiotic cannot improve performance and iron content in organs of broiler chicks and laying hens.
Oh Sin Geun;Yang Kwang Mo;Hur Won Joo;Yoo Young Hyun;Suh Hong Suk;Lee Hyung Sik
Radiation Oncology Journal
/
v.20
no.4
/
pp.367-374
/
2002
Purpose : To investigate the growth inhibitory effects, and the underlying mechanism of human colon cancer cell (HT-29) death, induced by a new synthetic bile acid derivative (HS-1200). Materials and Methods : Human colon cancer cells (HT-29), in exponential growth phase, were treated with various concentrations of a new synthetic bile acid derivative (HS-1200). The growth inhibitory effects on HT-29 cells were examined using a frypan blue exclusion assay. The extent of apoptosis was determined using agarose gel electrophoresis, TUNEL assays and Hoechst staining. The apoptotic cell death was also confirmed by Western blotting of PARP, caspase-3 and DNA fragmentation factor (DFF) analysis. To investigate the involvement of mitochondria, we employed immunofluorescent staining of cytochrome c and mitochondrial membrane potential analyses. Results : The dose required for the half maximal inhibition $(IC_{50})$ of the HT-29 cell growth was $100\~150\;{\mu}M$ of HS-1200. Several changes, associated with the apoptosis of the HT-29 cells, were reveal by the agarose gel eletrophoresis, TUNEL assays and Hoechst staining, following their treatment with $100\;{\mu}M$ of HS-1200. HS-1200 treatment also induced caspase-3, PARP and DFF degradations, and the western blotting showed the processed caspase-3 p20, PARP p85 and DFF p30 and p11 cleaved products. Mitochondrial events were also demonstrated. The cytochrome c staining indicated that cytochrome c had been released from the mitochondria in the HS-1200 treated cells. The mitochondrial membrane potential $(\Delta\Psi_m)$ was also prominently decreased in the HS-1200 treated cells. Conclusion : These findings suggest that the HS-1200 - induced apoptosis of human colon cancer cells (HT-29) is mediated via caspase and mitochondrial pathways.
In order to study the regional distribution and relative frequency of the immunoreactive endocrine cells in the gastrointestinal tract of the Mongolian gerbil, Meriones unguiculatus, the gastrointestinal tract was divided into 9 portions (cardia, fundus, pylorus, duodenum, jejunum, ileum, cecum, colon and rectum) and immunostained by immunohistochemical (PAP) method using 8 types of specific antisera against cholecystokinin (CCK)-8, gastrin, secretin, pancreatic polypeptide(PP), somatostatin, serotonin, glucagon and insulin. CCK-8-, gastrin-, somatostatin- and serotonin-immunoreactive cells were demonstrated in this study. These immunoreactive cells were found in the gastric gland regions of the pylorus or between parietal and chief cells of the fundus with round to spherical shaped, and in the interepithelial regions of the intestinal tract with spherical to spindle shaped except for jejunum where some of immunoreactive cells were also observed in the intestinal glands with round to spherical shaped. CCK-8-immunoreactive cells were restricted to the pylorus and duodenum with numerous and a few frequency, respectively. Gastrin-immunoreactive cells were restricted to the pylorus with numerous frequency. Similar to those of gastrin-immunoreactive cells, somatostatin-immunoreactive cells were restricted to pylorus with moderate frequency. Serotonin-immunoreactive cells were detected throughout whole gastrointestinal tract except for cardia and cecum with moderate or numerous frequency. However, no secretin-, PP-, glucagon- and insulin-immunoreactive cells were observed in this study. From these results, the appearance type, regional distribution and relative frequency of immunoreactive endocrine cells in the gastrointestinal tract of the Mongolian gerbils were somewhat lowered or restricted compared to those of other mammals and these differences were might be caused by feeding habits and species specification.
Background: Tissue hypoxia is characteristic of many human malignant neoplasm, and hypoxia inducible factor-1(HIF-1) plays a pivotal role in essential adaptive response to hypoxia, and activates a signal pathway for the expression of the hypoxia-regulated genes, resulting in increasing $O_2$ delivery or facilitating metabolic adaptation to hypoxia. Increased level of HIF-$1{\alpha}$ has been reported in many human malignancies, but in non-small cell lung carcinoma the influence of HIF-$1{\alpha}$ on tumor biology, including neovascularization, is not still defined. In present study the relationship of HIF-$1{\alpha}$ expression on angiogenetic factors, relationship between the tumor proliferation and HIF-$1{\alpha}$ expression, interaction of HIF-$1{\alpha}$ expression and p53, and relationship between HIF-$1{\alpha}$ expression and clinico-pathological prognostic parameters were investigated. Material and Method: Archival tissue blocks recruited in this study were retrieved from fifty-nine patients with primary non-small cell lung carcinoma, who underwent pneumonectomy or lobectomy from 1997 to 1999. HIF-$1{\alpha}$, VEGF(vascular endothelial growth factor), and p53 protein expression and Ki-67 labeling index in tumor tissues were evaluated, using a standard avidin-biotin-peroxidase complex(ABC) immunohistochemistry. Relationship between the HIF-$1{\alpha}$ expression and VEGF, p53 overexpression and correlation between the HIF-$1{\alpha}$ expresseion and Ki-67 index were analyzed. Clinico-pathologic prognostic parameters were also analyzed. Result: HIF-$1{\alpha}$ expression in cancer cells was found in 24 of 59 cases of non-small cell lung carcinoma(40.7%). High HIF-$1{\alpha}$ expression was significantly associated with several pathological parameters, such as pathological TMN stage(p=0.004), pT stage(p=0.020), pN stage (p=0.029), and lymphovascular invasion(p=0.019). High HIF-$1{\alpha}$ expression was also significantly associated with VEGF immunoreactivity(p<0.001), and aberrant p53 expression(p=0.040). but was marginally associated with Ki-67 labeling index(p=0.092). The overall 5-year survival rate was 42.3%. The survival curve of patients with a high HIF-$1{\alpha}$ expression was worse than that of patients with low-expression(p=0.002). High HIF-$1{\alpha}$ expression was independent unfavorable factors with a marginal significance in multivariate analysis performed by Cox regression. Conclusion: It is suggested that high HIF-$1{\alpha}$ expression may be associated with intratumoral neovascularization possibly through HIF-VEGF pathway, and high HIF-$1{\alpha}$ expression could be associated with lymph node metastasis and post operative poor prognosis in patients with non-small cell lung carcinoma.
Purpose: This study was conducted to determine the incidence, causative pathogens, risk factors and mortality for early onset sepsis in the first three days in very low birth weight infants. Methods: The medical records of 1,124 very low birth weight infants admitted to the neonatal intensive care unit of Samsung Medical Center between November 1994 and December 2008 were retrospectively reviewed. The incidence, causative pathogens, risk factors, and mortality for early onset sepsis in the first 3 days of life in very low birth weight infants were evaluated. Results: Early onset sepsis, as confirmed by positive blood cultures, was present in 17 of 1,124 infants (1.5%). Sixty-four percent of the isolated pathogens were gram-positive bacteria and 35% of the isolated pathogens were gram-negative bacteria. The dominant pathogens of early onset sepsis included Staphylococcus aureus (23.5%), Esherichia coli (23.5%), and Enterococcus (17.6%). Vaginal delivery (adjusted odds ratio [OR], 3.7; 95% confidence interval [CI], 1.3-10.3; P=0.01) was associated with early onset sepsis. The overall mortality (adjusted hazard ratio, 3.0; 95% CI, 1.4-6.5; adjusted P=0.0039) and mortality within 72 hours of life (adjusted hazard ratio, 6.5; 95% CI, 2.2-18.9; adjusted P=0.0005) of infants with early onset sepsis were higher than that of uninfected infants. Conclusion: Early onset sepsis remains an uncommon, but potentially lethal problem among very low birth weight infants. Knowledge of the likely causative organisms and risk factors for early onset sepsis can aid in instituting prompt and appropriate therapy, in order to minimize mortality.
Cho, Kyung-Soon;Park, Sun-Mee;Kim, Seang-Joon;Jung, Myung-Ju;Lee, Joo-Yeon;Kang, Chun
Korean Journal of Microbiology
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v.43
no.3
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pp.173-178
/
2007
The occurrence of acute respiratory infections caused by the influenza virus are particularly high during the winter season in Busan, Korea. In 2004 and 2005, a study of the rate of occurrences of the influenza virus was conducted. The results reveal that in 2004, of the 1,869 people with an acute respiratory infection that 154 (8.2%) people were infected by the influenza virus. In 2005, of the 1,579 people infected with an acute respiratory infection that 19 people (1.2%) were infected with the influenza virus. The study shows a decrease in the numbers of an influenza virus infection from 2004 to 2005. Data was collected by inspecting throat swabs and nasal discharge from those with an acute respiratory infection. Further inspection of the throat swab and nasal discharge from the infected individuals during 2004 and 2005 study show the occurrence of the different types of influenza virus in the population: 6 cases (3.5%) of influenza type A/H1N1, 129 cases (74.5%) of A/H3N2, and 38 cases (22.0%) of type B. The study conducted in 2004 and 2005 reveal that children between the ages of two and five were more likely to be infected than any other age group. In the study, about 62.2% of the infected individuals were between two and five years old. The detection rates between males and females are similar. However, it is notable that females are slightly more likely to develop an acute respiratory infection caused by the influence virus compared to their male counterparts.
A Pn10 DNA probe was introduced as a Prevotella nigrescens ATCC $33563^T$-specific DNA probe. In that study, the specificity of the Pn10 was tested with only type or reference strains of 5 oral bacterial species. The purpose of this study is to evaluate the specificity of the Pn10 using the wild type strains of P. nigrescens and is to develop the P. nigrescens ATCC $33563^T$-specific PCR primers based on the nucleotide sequence of the Pn10. The specificity of the Pn10 DNA probe was determined by Southern blot analysis. The nucleotide sequence of Pn10 DNA probes was determined by chain termination method. The PCR primers were designed based on the nucleotide sequence of cloned DNA fragment. The data showed that Pn10 DNA probe were hybridized with the genomic DNAs from P. nigrescens ATCC $33563^T$ and KB6. The Pn10 homologous region, KB6-Pn10, of P. nigrescens KB6 was cloned by PCR and sequenced. The Pn10 and KB6-Pn10 DNA fragments were consisted of 1,875 bp and 1,873 bp, respectively. The percent identity of the two was 98.8% and the divergence of them was 0.6%. The two primer sets (Pn10-F-AC/ Pn10-R-AC and Pn10-F-A/ Pn10-R-A), designed base on the nucleotide sequences of Pn10 DNA probe, were specific to the P. nigrescens ATCC $33563^T$. The two PCR primer sets could detect as little as 4 pg of genomic DNA of P. nigrescens ATCC $33563^T$. These results indicate that the two PCR primer sets have proven useful for the identification of P. nigrescens ATCC $33563^T$, especially with regard to the maintenance of the strain.
Ecological variations of Arbuscular mycorrhizal (AM) fungi were determined from the soils collected from different sites of the polluted and unpolluted areas related to the soil pollutions. Average 24.5 spores $(per\;20g)^{-1}$ soil of AM fungal spore were counted from the 32 sites of soils collected from On-san (polluted), whereas average 4.1 spores $(per\;20g)^{-1}$ soil from the 18 sites of those from Mt. Kwanak (polluted); Average 23.6 spores $(per\;20g)^{-1}$ soil of AM fungal spore were also counted from the 30 sites of soils collected from Chung-mu (unpolluted), whereas average 15.8 spores $(per\;20g)^{-1}$ soil from the 14 sites of those from Mt. Chungwang (unpolluted).The spores of AM fungi were the species of Glomus, Gigaspora, Acaulospora and Scutellospora. Among the above four genera, the species of Glomus were observed to be more abundant than the other genera in the soils collected from On-san, Chung-mu and Mt. Chungwang whereas the species of Gigaspora in those from Mt. Kwan-ak. The parameters of soils measured showed some variations between the polluted and unpolluted areas; 12.9 to 16.4% in the soil moisture, 5.6 to 8.3% in the organic matter and 4.3 to 5.7 at soil pH (polluted to unpolluted areas, respectively). The soils collected, thereby, appeared to be more strongly acidic and also lower in the contents of soil moisture or organic matter at the polluted area than unpolluted area. Based on the ecological criteria, the species richness or species diversity had significant differences (p<0.05) between polluted and unpolluted area. The spore density of genus Glomus or Gigaspora was significantly different (p<0.05) among the soils of three different plant vegetations (conifer plants, broad leaf plants, and grass plants). Also, there were significant differences (p<0.05) in the species evenness or species diversity among the soils referring to three different plant vegetations. There was a direct relationship $(r^2=0.38)$ between soil moisture and organic matter measured from 94 soil samples. Since there was a direct relationship $(r^2=0.22)$ between organic matter and total spores, it seems to be likely to presume that mycorrhizal spores can be increased in proportion to enhanced organic matters in soils. The species richness or species diversity was inclined to increase in proportion to enhanced soil pH and total spores in soils.
The studies were carried out to obtain the basic data for maximizing the protoplast yields from the mycelia of P. ostreatus and P. sajor-caju. Some factors affecting the regeneration of the protoplast of both species and the productivity of their reversion were also examined. The maximum yields of protoplasts were obtained from four days cultured mycelia of both species on cellophan membrane placed on the surface of PSA or MCM media in a petri dish. The optimal concentration of lytic enzyme Novozym 234 for protoplast releasing was 5 mg per ml of 0.5 M phosphate buffer solution with 0.6 M sucrose or 0.6 M $MgSO_4$ at pH 6.0. The greatest number of protoplasts was released 3 hours after incubation of the mycelia of P. ostreatus and after 4 hours for the P. sajor-caju in the lytic enzyme solution. Among the osmotic stabilizer solutions tested 0.6 M sucrose and 0.6 M KCl showed the best regeneration rates of the protoplasts of both species. When 0.75 % agar solution was over-layed on the regeneration media immediately after inoculation of the protoplast the regeneration rates were greatly enhanced. The ampicillin added to the agar solution prevented bacteria from infection. The reverted isolates produced the sporophores and basidial spores just like their parents without any mutations when they were cultivated in a broad mouth bottle with sawdust substrates.
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