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The Synergistic Effect of Juglans sinensis, Psoralea corylifolia, and Their Combination on the Asthmatic Murine Model (천식모델을 이용한 호도, 파고지 및 두 배합약물의 실험적 연구)

  • Kwon, Taek-Hyun;Lee, Young-Cheol;Lee, Jang-Cheon
    • The Korea Journal of Herbology
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    • v.24 no.2
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    • pp.39-48
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    • 2009
  • Objectives : To clarify the possible effect of JS (Juglans sinensis), PCF (Psoralea corylifolia L.), and J+P(JS+PCF), we examined their influence on the development of pulmonary eosinophilic inflammation in the asthmatic murine model. Methods : All mice were immunized on two different days (21 days and 7 days before inhalational exposure) by intraperitonial injections of 0.2 ml alum-precipitated Ag containing 100 ${\mu}$g of OVA bound to 4 mg of aluminum hydroxide in PBS. Seven days after the second sensitization, mice were exposed to aerosolized ovalbumin for 30 minutes/day on 3 days/week for 8 weeks (at a flow rate of 250 L/min, 2.5% ovalbumin in normal saline) and, JS, PCF and J+P (200 mg/kg, 400 mg/kg) were orally administered 3 times per week for 8 weeks. Results : The suppressive effects of JS, PCF, and J+P were demonstrated by the accumulation of eosinophils into airways, with the reduction of eosinophils and lung leukocytes. These were correlated with the marked reduction of IL-4, IL-5, IL-13 levels in the BALF and serum. OVA-specific IgE levels were also decreased in serum and BAL from these mice. And also JS, PCF, and J+P decreased eosinophilic CCR3 and CD11b expression in lung tissue. Conclusions : These results indicate that JS, PCF, and J+P have deep inhibitory effects on airway inflammation and hyper-responsiveness in the asthmatic murine model. The suppression of IL-5, IgE, and eosinophilils and the increase of IFN-${\gamma}$ production in BALF seem to contribute to these effects. Specially, esosinophils and TNF-a in J+P combination group were significantly reduced in BALF and lung tissue. Hence, the results indicated that JS, PCF, and J+P could act as an immuno-modulator which possesses anti-inflammatory and anti-asthmatic property by modulating the imbalance between Th1 and Th2 cytokines.

T Cell Microvilli: Finger-Shaped External Structures Linked to the Fate of T Cells

  • Hye-Ran Kim;Jeong-Su Park;Won-Chang Soh;Na-Young Kim;Hyun-Yoong Moon;Ji-Su Lee;Chang-Duk Jun
    • IMMUNE NETWORK
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    • v.23 no.1
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    • pp.3.1-3.14
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    • 2023
  • Microvilli are outer membrane organelles that contain cross-linked filamentous actin. Unlike well-characterized epithelial microvilli, T-cell microvilli are dynamic similar to those of filopodia, which grow and shrink intermittently via the alternate actin-assembly and -disassembly. T-cell microvilli are specialized for sensing Ags on the surface of Ag-presenting cells (APCs). Thus, these finger-shaped microprotrusions contain many signaling-related proteins and can serve as a signaling platforms that induce intracellular signals. However, they are not limited to sensing external information but can provide sites for parts of the cell-body to tear away from the cell. Cells are known to produce many types of extracellular vesicles (EVs), such as exosomes, microvesicles, and membrane particles. T cells also produce EVs, but little is known about under what conditions T cells generate EVs and which types of EVs are released. We discovered that T cells produce few exosomes but release large amounsts of microvilli-derived particles during physical interaction with APCs. Although much is unanswered as to why T cells use the same organelles to sense Ags or to produce EVs, these events can significantly affect T cell fate, including clonal expansion and death. Since TCRs are localized at microvilli tips, this membrane event also raises a new question regarding long-standing paradigm in T cell biology; i.e., surface TCR downmodulation following T cell activation. Since T-cell microvilli particles carry T-cell message to their cognate partner, these particles are termed T-cell immunological synaptosomes (TISs). We discuss the potential physiological role of TISs and their application to immunotherapies.

Association Analysis of MUC5AC Promoter Polymorphism with Asthma (MUC5AC 프로모터의 유전자 다형성과 천식과의 연관성)

  • Han, Seon-Sook;Sung, Ji Hyun;Lee, Mi-Eun;Lee, Seung-Joon;Lee, Sung Joon;Kim, Woo Jin
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.3
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    • pp.235-241
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    • 2007
  • Background: Airway mucus hypersecretion plays an important role in the pathogenesis of asthma, and is associated with the induction of MUC5AC expression in airway secretion. The MUC5AC gene is highly polymorphic; however, there are few studies about the association between the polymorphisms of the MUC5AC gene and asthma susceptibility or asthma phenotypes. We have investigated the association of MUC5AC promoter polymorphisms with the risk of asthma and asthma phenotypes. Methods: We determined the genotypes of the MUC5AC promoter (-1274G>A) in 78 asthma patients and in 78 age, sex-matched control individuals in the Korean population. Genomic DNAs from blood were analyzed by PCR and RFLP (restriction fragment length polymorphism) determination. We examined $FEV_1$, total eosinophil count, serum IgE level, $PC_{20}$ and the presence of atopy (by a skin test) in asthma patients. Results: The mean age of the patients was $47.7{\pm}16.1$ years and 38.5% were men, and the mean $FEV_1$ was $84.4{\pm}22.3%$ of predicted in the asthma patients. The -1274G>A polymorphism of the MUC5AC promoter in asthma patients was not significantly different as compared with normal individuals (GG 57.7%, AG 34.6% and AA 7.7% in asthma patients vs. GG 56.4%, AG 38.5% and AA 5.1% in control subject, p = 0.752, Cod). Several clinical parameters in asthma patients such as $FEV_1$, total eosinophil count, serum IgE level, $PC_{20}$ and the presence of atopy, were not associated with the -1274G>A polymorphism of the MUC5AC promoter. Conclusion: The -1274G>A single nucleotide polymorphism (SNP) frequency of the MUC5AC promoter was not associated with asthma in a Korean population.

Studies on the Cellulase produced by Myriococcum of albomyces (Myriococcum albomyces가 생산하는 Cellulase에 관한 연구)

  • Chung, Dong-Hyo
    • Applied Biological Chemistry
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    • v.14 no.1
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    • pp.59-97
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    • 1971
  • As a study on the cellulase of Myriococcum albomyces the culture media for enzyme formation and properties of its crude preparation were investigated and the crude enzyme preparation was further fractionated. The results are summarized as follows: 1. Wheat bran solid culture produced stronger activities of cellulase than rice bran or defatted soy bean meal solid culture. 2. Shaking culture with wheat bran, rice bran or defatted soy bean meal produced higher cellulase activities than solid culture with the corresponding media. 3. The enzyme formation was higher at $45^{\circ}C$ than at $37^{\circ}C$ or $50^{\circ}C$ regardless of the kind of culture medium. 4. The formation of CMCase activity was more promoted by organic nitrogen source than inorganic nitrogen source. 5. The formation of cellulase activities were increased 1.5 to 3.0-fold by adding CMC, Avicel or cellulose powder as an inducer into 5% wheat bran basal medium. 6. Cellulase production using a tank culture procedure with addition of CMC or Avicel as an inducer was the highest at fifth day and thereafter decreased slightly. 7. The crude enzyme preparation showed pH optimum in 4.0 to 4.5, and pH stability in the range of 3.5 to 8.0. Optimum temperature for the activity was $65^{\circ}C$ which was higher than among other cellulases and it was stable at $60^{\circ}C$ for 120 minutes. 8. Dialyzed crude enzyme was activated by $Ca^{++}$ and $Mg^{++}$, but inhibited by $Hg^{++}$, $Cu^{++}$ and $Ag^{+}$. 9. Four different types of cellulase, i. e., fraction I, fraction II-a, fraction II-b, and fraction III were purified from the culture filtrate of Myriococcum albomyces through a sequence of ammonium sulfate fractionation, and elution chromatography on DEAE-Sephadex A-25, Amberlite CG-25 type 2 and hydroxyapatite columns. 10. These four cellulase fractions were showed to be homogenous by electrophoresis and ultracentrifugation and also gave a typical ultraviolet absorption spectrum of protein. 11. Four purified fraction showed different specificity toward substrates, fraction I has a stronger activity toward Avicel, cellulose powder, and gauze than that of other cellulase fractions. Fraction II-a had a powerful activity toward cellobiose but it was almost inactive agaisnt fibrous cellulose contrary to fraction I. On the contrary, the main component fraction II-b had a fairly higher activity on CMC and Avicel. Activity of fraction II-b toward cellobiose was about one-third of that of fraction II-a and activity on Avicel was lower than that of fraction I. Fraction III had a more powerful activity in decreasing viscosity of CMC. 12. Final hydrolysis products of fibrous cellulose by each fraction were cellobiose and glucose. Whereas oligosaccharides were predominant in the early stage of hydrolysis, prolonged reaction produced more glucose than cellobiose. Fraction I and fraction II-a acted synergically on Avicel. 13. Optimum pH for the activities of cellulase fraction I, fraction II-a, fraction II-b and fraction III were found to be 5.5, 5.0, 4.0 and $4.0{\sim}4.5$, respectively. These fractions were found to be stable in the range of pH $3.0{\sim}7.5$. 14. Optimum temperature for the activities of fraction I, fraction II-a, fraction II-b, and fraction III were $50^{\circ}C$, $55^{\circ}C$, $60^{\circ}C$ and $55^{\circ}C$, respectively. No less of activity was found by heating 120 minutes at $55^{\circ}C$ and fraction II-a was more stable than the others at $60^{\circ}C$. 15. Fraction I and fraction II-b were activated by $Ca^{++}$ and $Mg^{++}$ but inhibited by $Hg^{++}$ and $Ag^{+}$.

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Production and evaluation of PRRS resistant pigs (PRRS 저항성 유전형 자돈의 생산 및 평가)

  • Jeong, Chang-Gi;Khatun, Amina;Nazki, Salik;Lee, Sim-In;Kim, Tae-Hun;Kim, Kwan-Suk;Park, Choi-Kyu;Kim, Won-Il
    • Korean Journal of Veterinary Service
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    • v.42 no.1
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    • pp.1-7
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    • 2019
  • Porcine reproductive and respiratory syndrome (PRRS) is economically the most important and challenging disease in swine industries worldwide and caused by PRRS virus (PRRSV). Previous studies reported that pigs with heterozygous genotypes in the guanylate-binding proteins (GBP1 and GBP5) exhibited increased resistance against PRRSV infection. The present study was conducted to produce higher numbers of the heterozygous pigs based on the PRRS resistant polymorphisms found in GBP1 (GBP1E2 and WUR) and GBP5, and evaluate the resistance of heterozygous pigs against challenge with a type 2 PRRSV (JA142) in comparison with homozygous pigs. In the challenge study, 12, 4 week-old PRRSV-negative piglets were selected based on the genotypes of the 3 polymorphisms (GBP1E2, WUR and GBP5). Among them, 8 piglets [homozygous (n=4) and heterozygous (n=4)] were challenged with JA142 and kept in the same room, and the remaining 4 piglets were kept separately as a negative control. In results, the sperms collected from the boars of GBP1E2-GG genotype produced approximately 28~41% higher numbers of heterozygous piglets as compared with those from the boars of GBP1E2-AG genotype. In the challenge study, we found that heterozygous piglets showed the significantly lower levels of viremia than homozygous piglets at 14, 21 and 28 dpc. Consistently, these heterozygous piglets also exhibited significantly higher ADWG than homozygous piglets. Therefore, in the current study, selection of boars based on SNP markers could increase the production of PRRS resistant piglets and the PRRS resistant pigs were found to be more resistant to PRRSV infection.

Growth and Photocurrent Properties of CdIn2S4/GaAs Single Crystal Thin Film by Hot Wall Epitaxy (Hot Wall Epitaxy 법에 의한 CdIn2S4 단결정 박막의 성장과 광전류 특성)

  • Lee, Sang-Youl;Hong, Kwang-Joon;Park, Jin-Sung
    • Journal of Sensor Science and Technology
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    • v.11 no.5
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    • pp.309-318
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    • 2002
  • A stoichiometric mixture of evaporating materials for $CdIn_2S_4$ single crystal thin films was prepared from horizontal electric furnace. To obtain the single crystal thin films, $CdIn_2S_4$ mixed crystal was deposited on thoroughly etched semi-insulating GaAs(100) substrate by the Hot Wall Epitaxy (HWE) system. The source and substrate temperatures were $630^{\circ}C$ and $420^{\circ}C$, respectively. The crystalline structure of the single crystal thin films was investigated by the photoluminescence and double crystal X-ray diffraction (DCXD). The carrier density and mobility of $CdIn_2S_4$ single crystal thin films measured with Hall effect by van der Pauw method are $9.01{\times}10^{16}\;cm^{-3}$ and $219\;cm^2/V{\cdot}s$ at 293 K, respectively. The temperature dependence of the energy band gap of the $CdIn_2S_4$ obtained from the absorption spectra was well described by the Varshni's relation, $E_g(T)=2.7116\;eV-(7.74{\times}10^{-4}\;eV)T^2/(T+434)$. The crystal field and the spin-orbit splitting energies for the valence band of the $CdIn_2S_4$ have been estimated to be 0.1291 eV and 0.0248 eV, respectively, by means of the photocurrent spectra and the Hopfield quasi cubic model. These results indicate that the splitting of the ${\Delta}so$ definitely exists in the ${\Gamma}5$ states of the valence band of the $AgInS_2$/GaAs epilayer. The three photocurrent peaks observed at 10K areascribed to the $A_1$-, $B_1$-, and C1-exciton peaks for n = 1.

DNA Polymorphisms in SREBF1 and FASN Genes Affect Fatty Acid Composition in Korean Cattle (Hanwoo)

  • Bhuiyan, M.S.A.;Yu, S.L.;Jeon, J.T.;Yoon, D.;Cho, Y.M.;Park, E.W.;Kim, N.K.;Kim, K.S.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.6
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    • pp.765-773
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    • 2009
  • Sterol regulatory element binding factor 1 (SREBF1) and fatty acid synthase (FASN) genes play an important role in the biosynthesis of fatty acids and cholesterol, and in lipid metabolism. This study used polymorphisms in the intron 5 of bovine SREBF1 and in the thioesterase (TE) domain of FASN genes to evaluate their associations with beef fatty acid composition. A previously identified 84-bp indel (L: insertion/long type and S: deletion/short type) of the SREBF1 gene in Korean cattle had significant associations with the concentration of stearic (C18:0), linoleic (C18:2) and polyunsaturated fatty acids (PUFA). The stearic acid concentration was 6.30% lower in the SS than the LL genotype (p<0.05), but the linoleic and PUFA contents were 11.06% and 12.20% higher in SS compared to LL (p<0.05). Based on the sequence analysis, five single nucleotide polymorphisms (SNPs) g.17924G>A, g.18043C>T, g.18440G>A, g.18529G>A and g.18663C>T in the TE domain of the FASN gene were identified among the different cattle breeds studied. Among these, only g.17924 G>A and g.18663C>T SNPs were segregating in the Hanwoo population. The g.17924G>A SNP is a non-synonymous mutation (thr2264ala) and was significantly associated with the contents of palmitic (C16:0) and oleic acid (C18:1). The oleic acid concentration was 3.18% and 2.79% higher in Hanwoo with the GG genotype than the AA and AG genotypes, respectively (p<0.05), whereas the GG genotype had 3.8% and 4.01% lower palmitic acid than in those cattle with genotype AA and AG, respectively (p<0.05). Tissue expression data showed that SREBFI and FASN genes were expressed in a variety of tissues though they were expressed preferentially in different muscle tissues. In conclusion, the 84-bp indel of SREBF1 and g.17924G>A SNP of the FASN gene can be used as DNA markers to select Hanwoo breeding stock for fatty acid composition.

Purification and Characterization of $\beta$-Xylosidase B of Bacillus stearothemophilus No.236 Produced by Recombinant Escherichia coli. (재조합 균주 Escherichia coli가 생산하는 Bacillus stearothermophilus No.236 $\beta$-Xylosidase B의 정제 및 특성)

  • 장욱진;조쌍구;최용진
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.297-302
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    • 1998
  • $\beta$-Xylosidase B was produced by Escherichia coli HB101/pKMG12 carrying the xylB gene of Bacillus stearothermophilus No.236 on its recombinant plasmid. The $\beta$-xylosidase B produced was purified by ammonium sulfate fractionation, DEAE-Sepharose CL-6B, Sephacryl S-200 and Superdex 200 HR gel filtration. The purified enzyme showed the highest activity at pH 6.5 and 5$0^{\circ}C$. But, the enzyme was observed to be very sensitive to the pH and temperature of the reaction mixture. The enzyme was activated about 35% of its original activity in the presence of 1 mM of $Mn^{2+}$ but it was completely inhibited by $Ag^{+}$, $Cu^{2+}$and $Hg^{2+}$ions. In contrast with the $\beta$-xylosidase A, the B enzyme was found to have $\alpha$-arabinofuranosidase activity though the activity was fairly low compared with the $\alpha$-arabinofuranosidase produced from the arfI gene of the same Bacillus stearothermophilus. Therefore, $\beta$-xylosidase B is considered to be more suitable than $\beta$-xylosidase A at least for the biodegradation of arabinoxylan. The $K_{m}$ and V$_{max}$ values of the $\beta$-xylosidase B for o-nitrophenyl-$\alpha$-D-xylopyranoside were 6.43 mM and 1.45 $\mu$mole/min, respectively. Molecular mass of the enzyme was determind to be about 54 kDa by SDS-PAGE and 160 kDa by Superdex 200HR gel filtration, indicating that the functional $\beta$-xylosidase B was composed of three identical subunits.s.

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Transcriptional Regulation of a DNA Repair Gene in Saccharomyces cerevisiae

  • Jang, Yeon-Kyu;Sancar, Gwen-B.;Park, Sang-Dai
    • Proceedings of the Zoological Society Korea Conference
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    • 1998.10b
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    • pp.113-113
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    • 1998
  • In Saccharomyces cerevisiae UV irradiation and a variety of chemical DNA -damaging agents induce the transcription of specific genes, including several involved in DNA repair. One of the best characterized of DNA -damage inducible genes is PHRI, which encodes the apoenzyme for DNA photolyase. Basal-level and damage-induced expression of PHRI require an upstream activation sequence, UASPHRI. Here we report the identification of the UlvIE6 gene of S. cerevisiae as a regulator of UASPHRl activity. Surprisingly, the effect of deletion of UME6 is growth phase dependent. In wild-type cells PHRI is induced in late exponential phase, concomitant with the initiation of glycogen accumulation that precedes the diauxic shift. Deletion of UNIE6 abolishes this induction, decreases the steady-state concentration of photolyase molecules and PHRI mRNA, and increases the UV sensitivity of a rad2 mutant. The results suggest that UM E6 contributes to the regulated expression of a subset of damage-responsive genes in yeast. Furthermore, the upstream repression sequence, URSPHRI, is required for repression and damage-induced expression of PHRl. Here we show identification of YER169W and YDR096W as putative regulators acting through $URS_{PHRI}$. These open reading frames were designated as RPHI (YERl69W) and RPH2 (YDR096W) indicating regulator of PHRI. Simultaneous disruption of both genes showed a synergistic effect, producing a four-fold increase in basal level expression and a similar decrease m the induction ratio following treatment of methyl methanesulfonate(MMS). Mutation of the sequence ($AG_4$) bound by Rphlp rendered the promoter of PHRI insensitive to changes in RPHI or RPH2 status. The data suggest that RPHI and RPH2 act as damage-responsive negative regulators of PHRI. Surprisingly, the sequence bound by Rphlp in vitro is found to be $AG_4$ which is identical to the consensus binding site for the regulators Msn2p and Msn4p involved in stress-induced expression. Deletion of MSN2 and MSN4 has little effect on the induction$.$ ratio following DNA damage. However, all deletions led to a significant decrease in basal-level and induced expression of PHRI. These results imply that MSN2 and MSN4 are positive regulators of P HRI but are not required for DNA damage repression. [Supported by grant from NIH]om NIH]

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Development of a gridded crop growth simulation system for the DSSAT model using script languages (스크립트 언어를 사용한 DSSAT 모델 기반 격자형 작물 생육 모의 시스템 개발)

  • Yoo, Byoung Hyun;Kim, Kwang Soo;Ban, Ho-Young
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.20 no.3
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    • pp.243-251
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    • 2018
  • The gridded simulation of crop growth, which would be useful for shareholders and policy makers, often requires specialized computation tasks for preparation of weather input data and operation of a given crop model. Here we developed an automated system to allow for crop growth simulation over a region using the DSSAT (Decision Support System for Agrotechnology Transfer) model. The system consists of modules implemented using R and shell script languages. One of the modules has a functionality to create weather input files in a plain text format for each cell. Another module written in R script was developed for GIS data processing and parallel computing. The other module that launches the crop model automatically was implemented using the shell script language. As a case study, the automated system was used to determine the maximum soybean yield for a given set of management options in Illinois state in the US. The AgMERRA dataset, which is reanalysis data for agricultural models, was used to prepare weather input files during 1981 - 2005. It took 7.38 hours to create 1,859 weather input files for one year of soybean growth simulation in Illinois using a single CPU core. In contrast, the processing time decreased considerably, e.g., 35 minutes, when 16 CPU cores were used. The automated system created a map of the maturity group and the planting date that resulted in the maximum yield in a raster data format. Our results indicated that the automated system for the DSSAT model would help spatial assessments of crop yield at a regional scale.