Kim, Chul Hwan;Kim, Hye Soo;Kim, Hong Chul;Kwon, Hyun Sook;Cheong, Jong-Chun;Kong, Won-Sik;Cho, Soo Jeong
Journal of Mushroom
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v.13
no.4
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pp.314-318
/
2015
This study was carried out to investigated the avaiability of dried Koojongsi persimmon peels (KPP) as a useful mushroom medium using Pleurotus eryngii ASI 2312. Mushroom cultivation medium used in this study was mixed with medium mixture, corn cob and sawdust (220:65:15, v/v). Dried KPP was replaced mushroom cultivation medium (control) with 5, 10, 15, 20, 30, 40, 50% dried KPP. The T-N content of dried KPP treatments decreased to increase replaced ratio of the dried KPP and C/N ratio was increased to increase replaced ratio of the dried KPP. But T-C content of dried KPP treatments was similar to untreated control. The average cultivating periods of mycelium on dried KPP treatments was delayed to increase replaced ratio of the dried KPP and cultivating periods was delayed over 30% dried KPP treatments. The length of stipe of dried KPP treatments was longer than that of the untreated control to increase replaced ratio of the dried KPP and thickness of stipes was tend to be thinner than that of the untreated control to increase replaced ratio of the dried KPP. The moisture, carbohydrate, crude protein and crude ash content of mycelial were similar to untreated control, but crude fatty acid was increased to increase replaced ratio of the dried KPP. The ${\beta}-glucan$ content of 10% and 15% treatments were higher than untreatment control. The results based on cultivation yield and ${\beta}-glucan$ content indicated that optimal mixture ratio dried KPP was 15%.
Vitellogenin(Vg) is a sex specific serum protein present in sexually maturing female blood of oviparous vertebrates. Estrogen($E_2$) is a main inducer of hepatic Vg synthesis. We investigated the effects of androgen and growth hormone(GH) on regulation of Vg and estrogen receptor(ER) genes in Japanese eel. Immature eels($200{\sim}250\;g$) were given a single injection of $E_2(5{\sim}5,000\;{\mu}g/kg\;bw)$ alone, or in combination with eel recombinant GH(eGH, $1{\sim}10\;{\mu}g/kg$) or methyltestosterone(MT, $1{\sim}5\;mg/kg$) and sacrificed 10 days after the hormone treatments. Expression levels of ER and Vg genes from the liver were determined by means of reverse transcription and polymerase chain reaction(RT-PCR). Administration of $E_2$ stimulated Vg gene expression in a dose dependent manner. Levels of Vg mRNA after the injection of $E_2(500\;{\mu}g/kg)$ with MT(5mg/kg) or eGH($10\;{\mu}g/kg$) were much higher than in that of $E_2$ alone($500\;{\mu}g/kg$). Whereas, injection of either vehicle, eGH ($10\;{\mu}g/kg$) or MT(5mg/kg) alone did not induce the expression of Vg gene in the liver. ER mRNA was detected from the fish treated with vehicle alone. $E_2$ injection($5{\sim}500\;{\mu}g/kg\;bw$) increased this ER expression but dose dependent response was not clear. Addition of MT(5mg/kg) or eGH($10\;{\mu}g/kg$) did not affect $E_2-stimulated$ ER mRNA expression. This study confirms the necessity of $E_2$ as the primary factor for Vg gene expression and requirement of additional hormones such as MT or GH for the full expression of Vg mRNA, and suggests that the additive effect of MT or GH on Vg gene expression would be mediated by some unknown factors other than ER.
Sporulation in the fission yeast Schizosaccharomyces pombe has been regarded as an important model of cellular development and differentiation. S. pombe cells proliferate by mitosis and binary fission on growth medium. Deprivation of nutrients especially nitrogen sources, causes the cessation of mitosis and initiates sexual reproduction by matting between two sexually compatible cell types. Meiosis is then followed in a diploid cell in the absence of nitrogen source. DNA fragment complemented with the mutations of sporulation gene was isolated from the S. pombe gene library constructed in the vector, pDB 248' and designated as pDB(spo5)1. We futher analyzed six recombinant plasmids, pDB(spo5)2, pDB(spo5)3, pDB(spo5)4, pDB(spo5)5, pDB (spo5)6, pDB(spo5)7 and found each of these plasmids is able to rescue the spo5-2, spo5-3, spo5-4, spo5-5, spo5-6, spo5-7 mutations, respectively. Mapping of the integrated plasmid into the homologous site of the S. pombe chromosomes demonstrated that pDB(spo5)1, and pDB(spu5)Rl contained the spo5 gene. Transcripts of spo5 gene were analyzed by Northern hybridization. Two transcripts of 3.2 kb and 2.5kb were detected with 5kb Hind Ⅲ fragment containing a part of the spo5 gene as a probe. The small mRNA(2.5kb) appeared only when a wild-type strain was cultured in the absence of nitrogen source in which condition the large mRNA (3.2kb) was produced constitutively. Appearance of a 2.5kb spo5-mRNA depends upon the function of the meil, mei2 and mei3 genes.
The varying characteristics between Korean's commercial and traditional soy sauces may be initiated by raw materials and fermentation techniques such as the koji and mezu process. We have examined properties of polysaccharides isolated from two different soy sauces which were made by the commercial process (CSP-0) and the traditional Korean process (KTSP-0) as well as their macrophage activities. Two polysaccharides have not effected the RAW 264.7 cells viability. The effects of CSP-0 and KTSP-0 on RAW 264.7 cells were demonstrated by the production of nitric oxide (NO), and reactive oxygen species (ROS). The CSP-0 and KTSP-0 significantly augmented NO and ROS productions by RAW 264.7 cells under a dose dependent manner. However, the activity of KTSP-0 was more potent than that of the CSP-0 at $1,000{\mu}g/m{\ell}$. The productions of IL-6 and TNF-${\alpha}$ were determined by real-time PCR and ELISA. mRNA expression levels of IL-6 and TNF-${\alpha}$ by KTSP-0 at $1,000{\mu}g/m{\ell}$ indicated 63 and 71 times higher than negative controls, respectively. Also, the production of IL-6 and TNF-${\alpha}$ by KTSP-0 at $1,000{\mu}g/m{\ell}$ showed 32.1 and 4.5 times higher than those by the CSP-0. To assess phagocytosis activities, the effects of CSP-0 and KTSP-0 on mRNA expression of Fc receptor I and II (FcR I, II) are being determined by RT-PCR products. Only the KTSP-0 showed enhanced expressions of mRNA expression for FcR I in a dose dependent manner, whereas the CSP-0 did not affect either the FcR I or II expressions. The above data lead us to conclude that the macrophage activations of Korean traditional soy sauce polysaccharide are higher than that of the commercial soy sauce polysaccharide.
Pollen germination viability is an essential factor to produce seeds from pollination and fertilization, which are required to maintain plant generation. In this study, we tried to identify the effect of boric acid on pollen germination and tube grouch in non-transgenic and transgenic plants expressing monoclonal antibodies (anti-colorectal cancer mAb CO17-1A, anti-breast cancer mAb BR55, and anti-rabies virus mAb57). The pollen of non-transgenic plant was treated with different concentration of boric acid (0, 5, 10, 15, 20, $40{\mu}g/mL$) in germination buffer to investigate its effect on in vitro pollen germination. At $20{\mu}g/mL$ of boric acid, tile pollen germination rate was the highest (49.5%) compared to other concentrations. In general, the germination rate significantly increased 3-10 folds in boric acid ($20{\mu}g/mL$) treated group in non-transgenic and transgenic plants. Also, the pollen tube length increased in boric acid ($20{\mu}g/mL$) treated groups. In the treated group, the pollen tube length increased until 3 h boric acid treatment and decreased after the 3 h, indicating that the 3 h is the most appropriate incubation time period. Western blot analysis showed that the mAb transgene expression was more stable in leaf than pollen in transgenic plants. This study suggested that $20{\mu}g/mL$ of boric acid is ideal concentration to induce in vitro pollen germination of transgenic plants expressing therapeutic monoclonal antibodies, indicating stable pollination and fertilization in transgenic plants.
Sin, Sang-Min;Cha, Jae-Young;Ha, Se-Eun;Sim, Sun-Mi;Kim, Hyoung-Do;Lee, Jung-Sup;Park, Jong-Kun
Journal of Life Science
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v.19
no.1
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pp.101-110
/
2009
The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel $mas2^+$ (mitosis associated protein) gene, a homolog of human SMARCAD1 was isolated and characterized from a fission yeast Schizosaccharomyces pombe (S. pombe) using gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 922 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that an SNF2 domain is located, which is involved in the chromosome remodeling. The quantitative analysis of the $mas2^+$ transcript against $adh1^+$ showed that the expression level of $mas2^+$ is high before septum formation in S. pombe. When $mas2^+$ null mutant cells were grown at 27 and $35^{\circ}C$, the cytokinesis of $mas2^+$ null mutant was greatly delayed and a large number of multi-septate and mis-segregated cells were produced. In addition, the number of multi-septate cells significantly increased. When cells were cultured in YES rich medium to increase proliferation, the abnormal phenotypes $mas2^+$ null mutant dramatically increased. These phenotypes could be rescued by an over-expression of the mast gene. The Mas2 protein localized in the nuclei of S. pombe, as evidenced by Mas2-EGFP signals. These results suggest that the $mas2^+$ is homologous to human SMARCAD1 gene and involved in septum formation and chromosome remodeling control.
Severe brain injuries induced by toxin pose one of the most important problems on our health care because of their high morbidity and mortality, are implicated to leucocyte infiltration more premature or immature brain than mature brain. Chemokines are the induction meditators for infiltration of inflammatory cells to the inflammation sites. In order to study the mechanism of leucocyte infiltration, the expression of several chemokines, MCP-1, $MIP-1{\alpha}$ and MIP-2 was studied in lipopolysaccharide(LPS)-stimulated neonatal and adult brain. One week old Sprague-Dawley rats or adult male rats weighing 300-350 g were used for the experiment. After anesthetization, $1\;{\mu}l$ LPS (0.5 mg/ml) subsequently was injected in the right caudate nucleus of the brain with stereotaxic frame. Animals were sacrificed at 6 hours, 24 hours, and 72 hours after injection. The present study was carried out using RT-PCR for the mRNA and immunohistochemistry for the expression of the proteins. In the neonatal rat brain, prominent interstitial edema with significant accumulation of leukocytes was detected at 24 and 72 hours after LPS injection. A semiquantitative analysis of RT-PCR revealed that the MCP-1, $MIP-1{\alpha}$, and MIP-2 mRNA expression peaked at 24 hours in neonatal and adult rat brain. Neonatal rats showed about 2.6, 1.4, and 1.2 times more expression of the MCP-1, $MIP-1{\alpha}$, and MIP-2 than that of the adult rats in the brain tissue. Immunohistochemical analysis also showed that MCP-1 immunoreactivity was paralleled with the RT-PCR results. MCP-1 protein was significantly detected at 24 and 72 hours in the brain parenchyma. $MIP-1{\alpha}$protein was highly expressed at 24 hours. The results of leukocyte infiltration in H&E stain was parallelled with that of the immunohistochemistry. Chemokine proteins were markedly detected at 24 hours after injection of LPS and neutrophil influx into intraparenchymal was prominent at 24 hours. These results suggest that the leukocyte infiltration in the intracranial infection may be controlled by mechanisms influenced by chemokine producing cells in the central nervous system such as microglia, astrocyte and endothelial cell.
In order to investigate the compositional change oil composts during the growing of cultivated mushroom (Agaricus bisporus), composts and mushrooms during the period of filling to ending under commercial conditions were subjected to chemical analyses. The results are summarized as follows and the mechanism of composting for mushroom cultivation was proposed. 1) The temperature change of growing bed and room was observed and the yield of mushroom for each cropping time was recorded to get $15.6kg/m^2$ in total crops. 2) Composts after filling showed pH 8.2 which dropped to 6.4 after casing and continued so up to ending. 3) On the dry weight basis of composts, crude ash increased whereas total nitrogen, ether extract and crude fibre decreased gradually to bring about the lowering of organic matter. 4) Total nitrogen of composts decreased gradually and more insoluble nitrogen was lost than soluble nitrogen. The C/N ratio of composts was initially 21 which was gradually lowered to 16. 5) The losses of ${\alpha}-cellulose$, pentosan and lignin in composts were 87%, 75%, and 60%, respectively, in which ${\alpha}-cellulose$ decreased markedly after casing. 6) Free reducing sugars of composts increased continuously. Gradually increased free amino acids till second cropping decreased again thereafter. Composts at the filling stage contained alanine, glutamic acid, glycine and serine in which glycine decreased markedly whereas proline increased remarkably upon mushroom cultivation. 7) Among minerals of composts, phosphorus and zinc tended to decrease, potassium and copper tended to increase anti sodium showed no marked change. 8) In comparison of mushrooms from different cropping time with respect to proximate composition, minerals, free reducing sugars and amino acids, no marked difference was observed. However, a little higher values were observed in crude fat, free reducing sugars and sodium content for early crops and in free amino acids and phosphorus content for late crops. Twelve free amino acids including alanine, serine, threonine, and glutamic acid were detected in the cultivated mushroom. 9) According to above experimental results, it was possible to support the mechanism of compositing that the formation of ammonia and decomposition of carbohydrates by mesophiles are followed by protein biosynthesis, formation of microbial bodies and nitrogen-rich lignin humus complex by thermophiles, thus supplying necessary nutrients for mushroom growth, along with residual carbohydrates.
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.
Pathways both mediated by and independent of transferrin(Tf) and the TfR have been described for the accumulation of iron. Although it is not clear whether the same systems take up iron and gallium, these pathways may suggest the contention that uptake of Ga-67 can, in fact, occur by both Tf-independent and Tf-dependent systems and may share with Fe-59 in part the same mechanism for uptake. The predominant system by which uptake of both radiometals occurs may be different in the degree of the transformation of tumor. Transformed(MMSV/3T3) and untransformed(BALB/3T3) cells were incubated with luM of Ga-67-citrate of Fe-59-chloride for 15 min. at $37^{\circ}C$ in either the presence or absence of Tf. After then, the monolayers were washed with HBSS or PBS, and the cells were solubilized in 1% SDS for gamma well counting and protein determinations. There were similarities, as well as differences, in the pattern of uptake of Fe-59 and Ga-67 presented both in ionic from and as bound to Tf. Both radiometals appeared gain to cells in either ionic or Tf-bound forms. Transformed cells appeared to accumulate more radiometal, either Ga-67 or Fe-59 in the presence of Tf than do the their untransforemd counterparts. Conversly the presentation of either radiometal in ionic form resulted in significantly greater accumulation of metal by the untransformed cells than those transformed. The efficiency for uptake of Ga-67 or Fe-59 in the absence of Tf was greater than for uptake of the Ga-Tf or Fe-Tf. However, the magnitude of difference in efficiency of uptake was greater for Fe-59(10-fold) than for Ga-67 (3-fold). Our results Supports the theory that both Tf-independent and Tf-dependent systems for the uptake of Ga-67 both systems operate oppositely between transformed cells and those untransformed, with uptake by the predominating in transformed cells by the Tf-mediated system and in untransformed cells by the Tf-independent. The uptake of Ga-67 by tumor may share with Fe-59 in part the same mechanism.
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