Rice blast caused by the filamentous fungus Magnaporthe oryzae, is arguably the most devastating rice disease worldwide. Development of a high-throughput and reliable field blast resistance evaluation system is essential for resistant germplasm screening, resistance genes identification and resistant varieties breeding. However, the occurrence of rice blast in paddy field is easily affected by various factors, particularly lack of sufficient inoculum, which always leads to the non-uniform occurrence and reduced disease severity. Here, we described a procedure for adequately inducing the occurrence of rice seedling blast in paddy field, which involves pretreatment of diseased straw, initiation of seedling blast for the first batch of spreader population, inducing the occurrence of the second batch of spreader population and test materials. This procedure enables uniform and consistent infection, which facilitates efficient and accurate assessment of seedling blast resistance for diverse rice materials.
Sungmin Hwang;Jung Up Park;Bohyun Yun;Ji-Won Park;WonWoo Lee
Proceedings of the Plant Resources Society of Korea Conference
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2023.04a
/
pp.12-12
/
2023
Plant biomass, or lignocellulose, is one of the most abundant natural resources on earth. Lignocellulosic biomass, such as agricultural and forestry residue, serves as a renewable feedstock for microbial cell factories due to its low price and abundant availability. However, the recalcitrance of lignocellulosic biomass requires a pretreatment process prior to microbial fermentation, from which fermentable sugars including xylose and glucose are generated along with various inhibitory compounds. The presence of furan derivatives, such as 5-hydroxymethyl-2-furaldehyde and 2-furaldehyde (furfural), hampers the microbial conversion of lignocellulosic biomass into value-added commodities. In this study, furfural tolerance was improved by investigating the detoxification mechanism in non-model yeast. The genes encoding aldehyde dehydrogenases were overexpressed to enhance furfural tolerance and resulted in improving cell growth and lipid production that can be converted into biofuel. Taken together, this approach contributes to the understanding of the reducing toxicity mechanism of furfural by the aldehyde dehydrogenases and provides a promising strategy that the use of microorganism as an industrial workhorse to treat efficiently lignocellulosic biomass as sustainable plant derivatives.
Microplastics in water resources have been recognized as a serious problem. The discharge of microplastics from wastewater treatment plants is considered a major contributor to environmental pollution in water resources. However, a reliable analytical method for quantifying microplastics in wastewater treatment plants has not yet been established. This study proposes a reliable, quick, and easy analytical method for quantifying microplastics. For the removal of organic particles, preprocessing steps were applied including oxidation, sonication, washing, and sieving. Nile Red staining was used to visualize microplastics, and quantitative analysis was conducted using fluorescent imaging. The stained microplastics were ultimately quantified through image analysis software. Among the preprocessing steps, sonication and washing stages were particularly effective in efficiently removing interfering substances from wastewater, enhancing the accuracy of the microplastic analysis. Additionally, various solvents (methanol, acetone, and N-hexane) for the Nile Red staining solution were tested. When N-hexane was applied as the solvent, the quantity of stained microplastics was lower compared to methanol and acetone. This suggests that N-hexane has a greater potential of reducing false staining and counting of non-plastic particles. In summary, this research demonstrates a robust method for quantifying microplastics in wastewater treatment plants by employing effective preprocessing steps and optimizing the staining process with Nile Red and N-hexane.
The study was conducted on 30 true acyclic Sahiwal cows (15 cows, ${\geq}90$ days postpartum; 15 postpubertal heifers, ${\geq}30$ months of age) and a similar 20 untreated controls (10 cows, 10 heifers). An 'Eazi' breed Controlled Internal Drug Release (CIDR) device (containing 1.38 g progesterone) was inserted intravaginally for 7 days (days 0 to 7) followed by 500 IU eCG i.m. at CIDR removal in all the treated animals. Heifers also received 5 mg oestradiol valerate i.m at CIDR insertion. The reproductive performance of these animals was recorded in terms of oestrus induction response, conception and pregnancy rates. Plasma progesterone ($P_4$) and oestradiol-$17{\beta}$ ($E_2$) profiles of 4 representative animals from each treatment group before, during and after CIDR treatment were also monitored. An oestrus induction response of 100% was observed in treated cows and heifers. The majority of cows (53.3%) and heifers (60%) were induced to oestrus within 24-36 and 36-48 h, respectively after CIDR withdrawal; with mean intervals of $44{\pm}3.18$ and $48{\pm}2.35h$, respectively. The conception rate at induced oestrus was higher in cows (40%) than heifers (20%). The final pregnancy rates after 2 subsequent oestruses were 80 and 60% in cows and heifers, respectively (overall 70% for all treated animals). In comparison, only 10% of control animals (2 cows only, 2/20) showed oestrus and become pregnant (10%) during theentire study period. The pretreatment (day 0) mean plasma P4 levels were statistically (p>0.05) similar in cows and heifers ($0.40{\pm}0.04$ and $0.49{\pm}0.11ng/ml$, respectively). The peak $P_4$ levels were observed on day 1 in cows ($13.94{\pm}1.41ng/ml$) and day 2 in heifers ($19.15{\pm}3.30ng/ml$) with a progressive decline up to the day of CIDR withdrawal ($3.35{\pm}0.92$ and $8.79{\pm}1.71ng/ml$, respectively). Mean $P_4$ levels on day 9 and 10 in cows and heifers did not differ significantly from their respective day 0 values and the lowest values were recorded on day 10 both in cows and heifers ($0.13{\pm}0.03$ and $0.14{\pm}0.02ng/ml$, respectively). Wide variations in individual pretreatment $E_2$ levels were observed both in the cows (range = 4-26, mean = $13.00{\pm}4.65pg/ml$) and heifers (range = 10-14, mean = $11.50{\pm}0.96pg/ml$). Thereafter also, $E_2$ levels in cows showed variation and reached a peak level ($53.50{\pm}2.99pg/ml$) on day 8. In heifers, peak mean $E_2$ level ($111.25{\pm}39.81pg/ml$) was recorded on day 1, followed by a non-significant decline on day 2, a significant fall on day 6 and a non-significant increase on day 9 and 10. However, mean $E_2$ levels on days 7 (p<0.05), 8 and 9 (p<0.01) were significantly higher in cows compared to heifers. The post-CIDR withdrawal mean highest $P_4$ and lowest $E_2$ levels coincided with the period when the majority of animals were induced to oestrus. CIDR and eCG treatment resulted in effective induction of oestrus with satisfactory pregnancy rates in true acyclic Sahiwal cows and heifers.
This study was conducted to establish optimized ${\beta}-glucan$ extraction method through enzymatic hydrolysis from Phellinus baumii and investigate ${\beta}-glucan$ contents and physicochemical properties. The optimal condition was obtained with the enzyme concentration of 0.66% (v/v), reaction time of 6.08 h ($R^2=0.9245$) and the ${\beta}-glucan$ contents from the Phellinus baumii extracts under the optimized condition was 1.9594 g/100 g. ${\beta}-Glucan$ yield (0.76-16.40%) of enzyme beta-glucan extract (EBE) was three fold higher than that of non-enzyme beta-glucan extract (NEBE). ${\beta}-Glucan$ purity (11.15-59.05%) of non-enzyme beta-glucan (NEB) and that of enzyme beta-glucan (EB) were higher than that of NEBE and that of EBE. ${\beta}-Glucan$ purity of EB (59.05%) and ${\beta}-glucan$ contents of EB (3.38 g/100 g) showed higher than those of others. Total sugar contents (0.61-1.17 mg/mL) showed that NEB and EB were higher than that of NEBE and EBE, EB had the highest total sugar content as 1.17 mg/mL, respectively. Protein contents (0.44-11.73 mg/mL) of NEBE and that of EBE were higher than that of NEB, that of EB. In FT-IR spectrum, the band at $890cm^{-1}$ of microcapsule was attributed to a ${\beta}-1,3-glucan$. The toxicities of ${\beta}-glucan$ from Phellinus baumii in both melanoma cell lines was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoli um bromide assay and ${\beta}-glucan$ from Phellinus baumii has no toxicity until $30{\mu}g/mL$. The effects of ${\beta}-glucan$ from Phellinus baumii on inhibition of cancer cell proliferation were detected by using a wound healing assay. The effect of NEB and EB were higher than NEBE and EBE, especially $30{\mu}g/mL$ of EB had the highest in both melanoma cell lines.
Potassium $(K^+)$ channels are present in airway smooth muscle cells, and their activation results in hyperpolarization and relaxation. Because these effects may have therapeutic relevance to hypersensitivity and asthma, we examined the effect of a potassium channel activator, cromakalim (BRL 34915, CK) on the release of mediators from superfused tracheal and parenchymal strips after passive sensitization with $IgG_1$ antibody. Both tissues were superfused with CK $(2{\times}10^{-6}\;M)$ for 30 min and challenged with CK and antigen (Ox-HSA). Using monodispersed, partially purified, highly purified guinea pig lung mast cells, we also examined the effect of CK on mediator release from these cells after passive sensitization with $IgG_{1}$ antibody $({\alpha}-OA)$. Guinea pig lung mast cells were purified using enzyme digestion method, count current elutriation, and discontinuous Percoll density gradient. After CK pretreatment, passively sensitized mast cells were challenged with varying concentration of antigen (OA, immunological stimuli) or with varying concentration of calcium ionophore (CaI, non-immunological stimuli). Histamine (Hist) release was determined by spectrophotofluorometry, and leukotrienes (LT) by radioimmunoassy. CK pretreatment decreased Hist by 35% and LT release by 40% in the antigen-induced tracheal tissue after $IgG_1$ sensitization but did not decrease the contractile response. In the antigen-induced parenchymal tissue CK decreased Hist release by 25% but poorly decreased LT. Both immunologic and non-immunologic stimuli caused a dose-dependent release of Hist and LT from monodispersed, partially purified and highly purified lung mast cells. Verification of LT release was obtained by the use of 5-lipoxygenase inhibitor, A64077 (Zileuton). CK decreased Hist and LT release by 20% respectively in the OA-induced guinea pig lung mast cells after $IgG_1$ sensitization. The inhibitory effects of CK on the Hist and LT release in the Ox-HSA-induced airway smooth muscle tissues or in the OA-induced and CaI-induced mast cells after $IgG_1$ sensitization were completely blocked by TEA and GBC. These studies show that guinea pig lung mast cells seem to be an important contributor to LT release, and that CK (which has been known as an airway smooth muscle relaxant) can in part act to inhibit mediator release in the antigen-induced airway smooth muscle, and that CK may also act to inhibit mediator release in the OA-induced and CaI-induced highly purified mast cells. These results suggest that Hist and LT release evoked by mast cell activation might in part be associated with $K{^+}4 channel activity.
Background : Neutrophils or monocytes separated in vitro by the adherence to plastic surface are known to be activated by surface adherence itself and subsequent experimental data might be altered by surface adherence. In the process of surface adherence, adhesion molecules have a clear role in intracellular signal pathway of cellular activation. Human alveolar macrophages(HAM) are frequently purified by the adherence procedure after bronchoalveolar lavage. But the experimental data of many reports about alveolar macrophages have ignored the possibility of adhesion-induced cellular activation. Method : Bronchoalveolar lavage was performed in the person whose lung of either side was confirmed to be normal by chest CT. With the measurement of hydrogen peroxide release from adherent HAM to plastic surface and non-adherent HAM with or without additional stimulation of phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine (fMLP), we observed the effect of the adherence to plastic surface. We also evaluated the effect of various biological surfaces on adhesion-induced activation of HAM. Then, to define the intracellular pathway of signal transduction, pretreatment with cycloheximide, pertussis toxin and anti-CD11/CD18 monoclonal antibody was done and we measured hydrogen peroxide in the culture supernatant of HAM. Results : 1) The adherence itself to plastic surface directly stimulated hydrogen peroxide release from human alveolar macrophages and chemical stimuli such as phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine(fMLP) colud not increase hydrogen peroxide release in these adherent macrophages which is already activated. 2) PMA activated human alveolar macrophages irrespective of the state of adhesion. However, fMLP stimulated the release of hydrogen peroxide from the adherent macrophages, but not from the non-adherent macrophages. 3) HAM adherent to A549 cell(type II alveolar epithelium-like human cell line) monolayer released more hydrogen peroxide in response to both PMA and fMLP. This adherence-dependent effect of fMLP was blocked by pretreatment of macrophages with cycloheximide, pertussis toxin and anti-CD18 monoclonal antibody, Conclusion : These results suggest that the stimulatory effect of PMA and fMLP can not be found in adherent macrophage because of the activation of human alveolar macrophage by the adherence to plastic surface and the cells adhered to biologic surface such as alveolar epithelial cells are appropriately responsive to these stimuli. It is also likely that the effect of fMLP on the adherent macrophage requires new protein synthesis via G protein pathway and is dependent on the adhesion between alveolar macrophages and alveolar epithelial cells by virtue of CD11/CD18 adhesion molecules.
In our previous studies, it was found that activities of maternal peripheral lymphocytes and thymocytes were depressed during the implantation period in rats and rabbits. This study was therefore attempted to clarify further this immunosuppression locally by determining lymphocyte response in lymph nodes draining the uterus (DLN) and to elucidate the mechanism by which prostaglandin E (PGE) modulates immune response during the implantation process in rats. As compared with non-pregnant rats, the response of DLN lymphocytes to concanavalin A (Con A) was depressed during the implantation period in 100% of rats studied. The activity of DLN lymphocytes depressed on day 8 of pregnancy was, however, restored partially by the treatment of indomethacin (ID), indicating that prostaglandin (PG) might be one of factors responsible for immunomodulation during the process of implantation. DLN lymphocyte activity in non-pregnant rats was suppressed if PGE was pre-treated prior to Con A and this suppression was partially restored by the treatment of ID. Furthermore, DLN lymphocytes pre-treated with PGE produced PGE in vitro and this PGE production was blocked by the treatment of ID, suggesting that PGE induced PGE-producing cells. However, the pretreatment of estradiol, progesterone, and hCG at doses enough to suppress lymphocyte activity was ineffective in inducing PGE-producing cells. From these results, it is suggested that PGE induces PGE-producing suppressor cells, thereby increasing PGE concentration and PGE in turn depresses maternal local immune response as well as systemic immune response during the implantation period in rats.
Background: Several prognostic factors have been studied in NSCLC, although it is unknown which is most useful. In this study, we aimed to investigate whether pre-treatment serum albumin level has prognostic value in patients with Stage IIIB NSCLC. Materials and Methods: This cross-sectional study included a total of 204 patients with Stage IIIB NSCLC who met the inclusion criteria. Pre-treatment serum albumin levels and demographic, clinical, and histological characteristics, as well as laboratory variables were recorded. A cut-off value was defined for serum albumin level and the patients were stratified into four groups on thios basis. Results: The majority of the patients was males and smokers, with a history of weight loss, and squamous histological type of lung cancer. The mean serum albumin level was $3.2{\pm}1.7g/dL$ (range, 2.11-4.36 g/dL). A cut-off value 3.11 g/dL was set and among the patients with a lower level, 68% had adenocarcinoma and 82% were smokers. The patients with low serum albumin levels had a lower response rate to e first-line chemotherapy with a shorter progression-free survival and overall survival. Multivariate analysis showed that low serum albumin level was an independent poor prognostic factor for NSCLC. Conclusions: This study results suggest that low serum albumin level is an independent poor prognostic factor in patients with Stage IIIB NSCLC, associated with reduction in the response rate to first-line therapy and survival rates.
Objective: To explore the relationships between primary tumor $^{18}F$-FDG uptake measured as the SUVmax and local extension, and nodal or distant organ metastasis in patients with NSCLC on pretreatment PET-CT. Methods: 93 patients with NSCLC who underwent $^{18}F$-FDG PET-CT scans before the treatment were included in the study. Primary tumor SUVmax was calculated; clinical stages, presence of local extension, nodal and distant organ metastases were recorded. The patients with SUVmax${\geq}2.5$ were divided into low and high SUVmax groups by using the median SUVmax. The low SUVmax group consisted of 45 patients with SUVmax<10.5, the high SUVmax group consisted of 46 patients with SUVmax${\geq}10.5$. Their data were compared statistically. Results: 91 cases with SUVmax${\geq}2.5$ were included for analysis. The mean SUVmax in patients without any metastasis was $7.42{\pm}2.91$ and this was significantly lower than that ($12.18{\pm}4.94$) in patients with nodal and/or distant organ metastasis (P=0.000). In the low SUV group, 19 patients had local extension, 22 had nodal metastasis, and 9 had distant organ metastasis. In the high SUV group, 31 patients had local extension, 37 had nodal metastasis, and 18 had distant organ metastases. There was a significant difference in local extension (P =0.016), distant organ metastasis (P =0.046), and most significant difference in nodal metastasis rate (P =0.002) between the two groups. In addition, there was a moderate correlation between SUVmax and tumor size (r = 0.642, P<0.001), tumor stage (r = 0.546, P<0.001), node stage (r = 0.388, P<0.001), and overall stage (r = 0.445, P= 0.000). Conclusion: Higher primary tumor SUVmax predicts higher extensional or metastatic potential in patients with NSCLC. Patients with higher SUVmax may need a close follow-up and more reasonable individual treatment because of their higher extensional and metastatic potential.
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