• Journal of Mushroom
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• v.9 no.1
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• pp.3-16
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• 2011

In the bipolar basidiomycete Pholiota nameko, a pair of homeodomain protein genes located at the A mating-type locus regulates mating compatibility. In the present study, we used a DNA-mediated transformation system in P. nameko to investigate the homeodomain proteins that control the clamp formation. When a single homeodomain protein gene (A3-hox1 or A3-hox2) from the A3 monokaryon strain was introduced into the A4 monokaryon strain, the transformants produced many pseudo-clamps but very few clamps. When two homeodomain protein genes (A3-hox1 and A3-hox2) were transformed either separately or together into the A4 monokaryon, the ratio of clamps to the clamp-like cells in the transformants was significantly increased to approximately 50%. We, therefore, concluded that the gene dosage of homeodomain protein genes is important for clamp formation. When the sip promoter was connected to the coding region of A3-hox1 and A3-hox2 and the fused fragments were introduced into NGW19-6 (A4), the transformants achieved more than 85% clamp formation and exhibited two nuclei per cell, similar to the dikaryon (NGW12-163 ${\times}$ NGW19-6). The results of real-time RT-PCR confirmed that sip promoter activity is greater than that of the native promoter of homeodomain protein genes in P. nameko. So, we concluded that nearly 100% clamp formation requires high expression levels of homeodomain protein genes and that altered expression of the A mating-type genes alone is sufficient to drive true clamp formation.

• Mycobiology
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• v.30 no.1
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• pp.31-36
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• 2002

Soil bacteria were screened for the ability to control cucumber anthracnose caused by Colletotrichum orbiculare through induced systemic resistance(ISR). Sixty-four bacterial strains having in vitro antifungal activity were used for selecting ISR-inducing strains in cucumber. Cucumber seeds(cv. Baeknokdadagi) were sown in potting mixtures incorporated with the soil bacteria, at a rate of ca. $10^8$ cells per gram of the mixture. Two week-old plants were then transplanted into the steam-sterilized soil. Three leaf-stage plants were inoculated with a conidial suspension($5{\times}10^5$ conidia/ml) of C. orbiculare. Diseased leaf area(%) and number of lesions per $cm^2$ leaf were evaluated on third leaves of the plants, $5{\sim}6$ days after inoculation. Among 64 strains tested, nine strains, GC-B19, GC-B35, GK-B18, MM-B22, PK-B14, RC-B41, RC-B64, RC-B65, and RC-B77 significantly(P=0.05) reduced anthracnose disease compared to the untreated control. In contrast, some bacterial strains promoted susceptibility of cucumber to the disease. From the repeated experiments using the nine bacterial strains, GC-B19, MM-B22, PK-B14, and RC-B65 significantly(P=0.05) reduced both diseased leaf area(%) and number of lesions per $cm^2$ leaf in at lease one experiment. These strains with control efficacy of $37{\sim}80%$ were determined to be effective ISR-inducing strains.

• YAKHAK HOEJI
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• v.37 no.6
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• pp.605-614
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• 1993

To investigate the endothelial dependence of angiotensin II(A II)-induced responses in the systemic and pulmonary arterial system of acute renal hypertensive rats of 2-kidney, 1-ligation type (RHRs), A II-induced vasocontractile and pressor effects were evaluated in isolated arteries and in vivo, respectively. A II dose-dependently contracted intact thoracic aorta and pulmonary artery (E$_{max}$:40% at 10$^{-7}$M and 80% at 3$\times$10 $^{-8}$M, respectively) from normotensive rats(NRs), which was significantly increased by removal of endothelial cells or pretreatment with EDRF inhibitors. In NRs, A II increased mean systemic and pulmonary arterial pressure(33 and 5.6mmHg at 0.1 $\mu\textrm{g}$/kg, respectively), the effect being significantly increased (P<0.01) by L-NAME(30mg/kg, i.v.). However, A II-induced contraction of intact thoracic aorta and pulmonary artery(E$_{max}$: 33% at 10$^{-7}$M and 93% at 3$\times$10$^{-8}$M, respectively) from RHRs were not changed after endothelial function was disrupted as above; similarly, pressor effects of A II on the systemic and pulmonary arterial pressure in RHRs did not altered by L-NAME. A II tachyphylactic responses for intact thoracic aorta from NRs and RHRs(65 and 87% at 10$^{-8}$M, respectively) were greater than those for pulmonary artery(19 and 19% at 10$^{-8}$M, respectively). Distruption of endothelial function significantly (P<0.01) depressed A II tachyphylaxis for thoracic aorta, but not for pulmonary artery. These results suggest that vascular reactivity to A II is not altered in RHRs, and it is greater for pulmonary arterial system than for systemic arterial system. A II reactivity is EDRF-dependent in both arterial systems of NRs, but EDRF-independent for RHRs. Finally, EDRF is one of the major factors underlying A II tachyphylaxis for thoracic aorta, but not for pulmonary artery.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.19 no.1
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• pp.54-60
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• 2016

Purpose: The aim of this study is to investigate the association between elevated alanine aminotransferase (ALT) and urosepsis in children with acute pyelonephritis (APN). Methods: We retrospectively identified all children who were managed in our hospital with APN during a decade period. In our study a diagnosis of APN was defined as having a positive urine culture and a positive (99m)Tc-dimercaptosuccinic acid scintigraphy. We compared those with elevated ALT and those with normal ALT according to the following variables: age, gender, duration of fever prior to admission, presence of hypotension, C-reactive protein (CRP), creatinine, presence of anemia, white blood cells count, platelet count, blood culture result, and grades of vesicoureteral reflux. In addition, the correlation between elevated ALT and positive blood culture was analyzed in detail. Results: A total of 996 children were diagnosed with APN, of which 883 were included in the study. ALT was elevated in 81 children (9.2%). In the analysis of demographic characteristics, the number of children with elevated ALT was higher in children between 0 to 3 months, boys, and in those with positive blood culture (p=0.002, 0.036, and 0.010, respectively). In multivariate analysis of variables associated with positive blood culture, age younger than 3 months, elevated ALT, elevated CRP, and elevated creatinine showed statistical significance (p=0.004, 0.030, 0.043, and 0.044, respectively). Conclusion: Our study demonstrates the association between elevated ALT and increased prevalence of urosepsis in addition to elevated CRP, elevated creatinine, and age younger than 3 months in children with APN.

• Korean Journal of Animal Reproduction
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• v.19 no.2
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• pp.81-88
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• 1995

Culture medium (ASF-301) of tHUE-2 cell, human endothelial cell line, and culture medium of these cells (conditioned medium : CM) which affect embryonic development of in vivo fertilized 1-cell embryos of mouse were examined. Two-cell stage block of mouse embryos was overicomed in ASF-301 and CM without EDTA, which usually added in basic medium (modified Whitten Medium: MWM, control) to overcome the 2-cell stage block. The developmental rates of embryos to the blastocyst stage were significantly increased in MWM containing 12.5% of growth factors added to ASF-301 (10mg/ $\ell$ transferrin, 1mg/$\ell$ insulin, 0.01mg/$\ell$ EGF) than those of 100% addition and control, 78.0% vs 20.8 and 52.3%, respectively (P<0.05), but the growth factors was not affected the hatching rate of blastocyst. Using ASF-301 or CM which was not treated, embryonic development into the blastocyst and hatched blastocyst stages were not affected. However, proportions of embryonic development into the blastocyst and hatched blastocyst stages were significantly higher in dilution (ASF-301 1:10; CM 1:3~1:6) than those in control (P,0.05). In ASF-301 dialyzed M.W.<10000 dialysis membrane, the developmental rate upto the hatched blastocyst stage was significantly increased, compared to ASF-301 which was not dialyzed (P<0.05), and hatching rate of blastocyst of these group was singnificantly increased than those in MWM (P<0.05). Compared to CM which was not dialyzed, however, in dialyzed CM was significantly decreased, compared to untreated CM (P<0.05), especially any hatched blastocyst was not appeared. As a result of these experiments indicated that a kind or porper treatment such as a dilution of complex synthetic cell culture medium and conditioned medium, and that a optimal concentration of growth factors are usuful for embryo cultrue in vitro.

• Molecules and Cells
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• v.19 no.3
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• pp.418-427
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• 2005

When inoculated into sensitive tobacco Xanthi-nn plants, the crucifer and garlic-infecting Tobacco mosaic virus (TMV-Cg) induces local necrotic lesions that resemble those seen in the hypersensitive response (HR) of resistant tobacco plants. However, unlike these, tobacco Xanthi-nn plants do not become resistant to infection and the virus spreads systemically causing a severe disease characterized by necrotic lesions throughout the plant. To identify the viral protein that elicits this necrotic response, we used a set of hybrid viruses constructed by combination of TMV-Cg and the tobacco mosaic virus strain U1 (TMV-U1). In this study we present evidence that the coat protein of TMV-Cg (CPCg) is the elicitor of the necrotic response in tobacco Xanthi-nn plants. Local and systemic necrotic lesions induced by TMV-Cg and by the hybrid U1-CPCg -that carries CPCg in a TMV-U1 context- are characterized by cell death and by the presence of autoflorescent phenolic compounds and $H_2O_2$, just like the HR lesions. In addition, defense-related genes and detoxifying genes are induced in tobacco Xanthi-nn plants after TMV-Cg and U1-CPCg inoculation. We postulate that in our system, CPCg is recognized by sensitive tobacco plants that mount an incomplete defense response. We call this an HR-like since it is not enough to induce plant resistance.

• KSBB Journal
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• v.16 no.2
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• pp.128-132
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• 2001

Cyclodextrin glucanotransferase (EC 2.4.1.19 : 1,4-$alpha$-glucan 4-$alpha$-D-(1,4-glucano) transferase, cyclizing; CGTase) was recovered by starch adsorption. The adsorption and desorption of CGTase to starch was studied as a function of pH, temperature, and starch type. The optimal pH, temperature, and starch for adsorption were, 8.0, $4^{circ}C$, and 1% (w/v) corn starch, respectively, per 205 U/mL enzyme activity in the presence of 25% (w/v) ammonium sulfate. The maximum adsorption ratio was 95%. On the other hand, the optimal pH, temperature, and starch type for desorption were 8.0 (tris-buffer), $50^{circ}C$, and oxidized starch, respectively. The maximum desorption ratio was 98% by tris-buffer solution at pH 8.0. The efficiency of adsorption and desorption were affected slightly by the removal of cells from the fermentation broth.

• IMMUNE NETWORK
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• v.1 no.3
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• pp.236-243
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• 2001

Background: An immunological approach for aging mechanism appears to be important. Lymphocyte subsets analysis in peripheral blood is widely performed to assess the immune status and to diagnose and monitor various diseases. Some lymphocyte subsets are known to change with age, but only few data about age-related reference ragnes for these subsets in healthy individuals have been reported. So we attempted to report reference ranges for these subsets in each age group and review changes of the results with age for the secondary studies about immune cell function as lymphocyte blast transformation and immunoglobulin gene rearrangement (VDJ) including recombination activating genes (RAG-1 and RAG-2). Methods: Lymphocyte subset analysis was performed on 302 subjects, 189 males and 113 females with age group of all decades of life. Two color direct immunofluorescene flow cytometry (FCM) was done using $Simultest^{TM}$ IMK-Lymphocyte kit (Becton Dickinson, USA), $FACScan^{TM}$ (Becton Dickinson, USA) and $FACSCalibur^{TM}$ (Becton Dickinson, USA). Lymphocyte subsets analysed were T ($CD3^+$) and B cells ($CD19^+$), helper/inducer T ($CD4^+$) and suppressor/cytotoxic T cells ($CD8^+$), helper/suppressor ($CD4^+/CD8^+$) ratio and natural killer (NK) cells ($CD3^-CD16^+/CD56^+$). The absolute numbers of each subset were calculated from total lymphocyte counts. Data collected was analysed using SAS 6.12. A P-value of < 0.05 was considered significant. Results: We reported the counts and percentages of lymphocyte and these subsets in each age group. There were no statistically significant differences between male and female subjects. The percentage of $CD4^+$ T cells, and the count of NK cells did not show the significant difference among the various age groups. The age-related changes observed in our study were as following: 1) a decrease in the percentages of T cells, B cells and $CD8^+$ T cells ; 2) a decrease in the counts of B cells and $CD8^+$ T cells ; 3) an increase in the percentage and count of NK cells ; and 4) an increase in the $CD4^+/CD8^+$ ratio. Conclusion: The characteristics of aging process appeared to be showing a marked decrease of lympocyte subsets T and B cells as well as T8 ($CD8^+$). The age-related increase of the percentage of cells bearing NK marker can be interpreted as a compensatory consequence to cope with the decrease of T cells related to the thymic involution. These changes with age appeared to be for the secondary study about immune cell function as lymphocyte blast transformation and immunoglobulin gene rearrangement.

• Tuberculosis and Respiratory Diseases
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• v.52 no.2
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• pp.107-116
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• 2002

Background: Matrix metalloproteinases(MMP) are essential enzymes for tumor invasion and metastasis. Among the MMP family, elevated MMP-9 and stromelysin-3(STR-3) expression have been reported to be poor prognostic factors in lung cancer patients. To evaluate the possibility of a molecular diagnosis of lung cancer using peripheral blood, the mRNA expression level of MMP-9 and STR-3 was measured using a reverse transcriptase-polymerase chain reaction (RT-PCR) in patients with lung cancer. Methods : Ninety six patients(44 patients with lung cancer, 19 pulmonary infection, and 33 control) were included. To detect MMP-9 and STR-3 mRNA expression, RT-PCR was performed in peripheral blood mononuclear cells. ELISA was also used to measure the serum level of MMP-9. Results : MMP-9 was expressed more frequently in patients with a pulmonary infection(18/19, 94.7%) compared to lung cancer patients(26/44, 59.1%) or the controls (23/33, 69.7%) (p=0.018). On the other hand, STR-3 expression was observed more frequently in patients with lung cancer(37/44, 84.1%) compared to the lung infection patients(8/19, 42.1%) or control(20/33, 60.6%) (p=0.003). Among the lung cancer patients, MMP-9 was expressed more frequently when a tumor invaded the lymph nodes(17/24, 70.8%) compared to when a tumor did not(3/13, 23.1%) (p=0.005). The MMP-9 and STR-3 expression levels had no relationship with age, sex, tumor size, distant metastasis, or tumor histology. The serum MMP-9 concentration was not higher in lung cancer patients compared to patients with a pulmonary infection or the control subjects. Conclusion : STR-3 may be used as a diagnostic marker in the peripheral blood of lung cancer patients using RT-PCR. Further studies to evaluate the clinical significance of elevated STR-3 expression in lung cancer patients is recommended.

• The Journal of Korean Medicine
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• v.30 no.2
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• pp.63-78
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• 2009

Objective: The purpose of experimental study was to prove the effects of Boyangmakseong-bang (BYMSB) treatment on cBSA-induced in a MN mouse model. Methods: We divided mice into 4 groups. The Normal group had no treatment. We used cBSA and induced MN mouse model to the other 3 groups. The Control group was treated with cBSA (9mg/kg i.p) only. The second group, named 'BY-250', was treated with cBSA (9mg/kg i.p) and BYMSB extract (250mg/kg, p.o). The third group, named 'BY-500', was treated with cBSA (9mg/kg i.p) and BYMSB extract (500mg/kg, p.o). After cBSA and BYMSB extract treatment for 4 weeks, the increase in percentage of body weight, proteinuria, serum albumin, total cholesterol, creatinine and BUN of all groups were measured. The CD3+, CD19+, CD4+, CD8+ cell levels of spleen of all groups were analyzed. IgA, IgG, IgM, IL-$1{\beta}$, TNF-${\alpha}$, IL-6 and IFN-${\gamma}$ levels of all groups were gauged. H&E staining, immunofluorescence staining and electron microscopy of kidney were observed. Results: BYMSB showed significant decrease in the 24hrs proteinuria, serum total cholesterol, serum IgG levels and BUN levels, and showed significant increase in the serum albumin levels compared with the control group. BYMSB showed increase in the increasing percentage of body weight and IFN-${\gamma}$ levels compared with the control. BYMSB showed decrease in the CD3+ T cells, CD4+ Th cells, IL-$1{\beta}$, TNF-${\alpha}$ and IL-6 levels, but did not show significant change compared with the control. BYMSB showed considerable decrease in the thickening of the GBM on H&E staining, deposition of IgG on immunofluorescence staining and deposition of electron-density on electron microscopy of kidney compared with the control. Conclusions: According to the above results, it is suggested that BYMSB decreases the symptoms of MN induced by cBSA in a mouse model. Therefore BYMSB seems to be applicable to MN in clinical practice.