• Parasites, Hosts and Diseases
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• v.61 no.4
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• pp.405-417
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• 2023

Chagas disease, caused by Trypanosoma cruzi parasite, is a significant but neglected tropical public health issue in Latin America due to the diversity of its genotypes and pathogenic profiles. This complexity is compounded by the adverse effects of current treatments, underscoring the need for new therapeutic options that employ medicinal plant extracts without negative side effects. Our research aimed to evaluate the trypanocidal activity of Bidens pilosa fractions against epimastigote and trypomastigote stages of T. cruzi, specifically targeting the Brener and Nuevo León strains-the latter isolated from Triatoma gerstaeckeri in General Terán, Nuevo León, México. We processed the plant's aerial parts (stems, leaves, and flowers) to obtain a methanolic extract (Bp-mOH) and fractions with varying solvent polarities. These preparations inhibited more than 90% of growth at concentrations as low as 800 ㎍/ml for both parasite stages. The median lethal concentration (LC₅₀) values for the Bp-mOH extract and its fractions were below 500 ㎍/ml. Tests for cytotoxicity using Artemia salina and Vero cells and hemolytic activity assays for the extract and its fractions yielded negative results. The methanol fraction (BPFC3MOH1) exhibited superior inhibitory activity. Its functional groups, identified as phenols, enols, alkaloids, carbohydrates, and proteins, include compounds such as 2-hydroxy-3-methylbenzaldehyde (50.9%), pentadecyl prop-2-enoate (22.1%), and linalool (15.4%). Eight compounds were identified, with a match confirmed by the National Institute of Standards and Technology (NIST-MS) software through mass spectrometry analysis.

• Journal of Microbiology and Biotechnology
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• v.33 no.11
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• pp.1437-1447
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• 2023

A recently bioinformatic analysis of genomic sequences of fungi indicated that fungi are able to produce more secondary metabolites than expected. Despite their potency, many biosynthetic pathways are silent in the absence of specific culture conditions or chemical cues. To access cryptic metabolism, 108 fungal strains isolated from various sites were cultured with or without Streptomyces sp. 13F051 which mainly produces trichostatin analogues, followed by comparison of metabolic profiles using LC-MS. Among the 108 fungal strains, 14 produced secondary metabolites that were not recognized or were scarcely produced in mono-cultivation. Of these two fungal strains, Myrmecridium schulzeri 15F098 and Scleroconidioma sphagnicola 15S058 produced four new compounds (1-4) along with a known compound (5), demonstrating that all four compounds were produced by physical interaction with Streptomyces sp. 13F051. Bioactivity evaluation indicated that compounds 3-5 impede migration of MDA-MB-231 breast cancer cells.

• FOCUS: LIFE SCIENCE
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• no.1
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• pp.3.1-3.7
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• 2024

The article discusses the critical role of chromatography in the analysis and purification of proteins in biopharmaceuticals, emphasizing the importance of comprehensive characterization for ensuring their safety and efficacy. It highlights the use of Avantor® ACE® HPLC columns for the separation and purification of proteins, focusing on the analysis of intact proteins using reversed-phase liquid chromatography (RPLC) with fully porous particles. This article also details the application of different mobile phase additives, such as TFA and formic acid, and emphasizes the advantages of using type B ultra-pure silica-based columns for efficiency and peak shape in biomolecule analysis. Additionally, it addresses the challenges of analyzing intact proteins due to slow molecular diffusion and introduces the concept of solid-core (or superficially porous) particles, emphasizing their benefits over traditional porous particles for the analysis of therapeutic proteins. Furthermore, it discusses the development of Avantor® ACE® UltraCore BIO columns, specifically designed for the high-efficiency separation of large biomolecules, such as proteins, and demonstrates their effectiveness in achieving high-resolution separations, even for higher molecular weight proteins like monoclonal antibodies (mAbs). In addition, it underscores the complexity of analyzing and characterizing intact protein biopharmaceuticals, requiring a range of analytical techniques and the use of wide-pore stationary phases, operated at elevated temperatures and with relatively shallow gradients. It highlights the comprehensive range of options offered by Avantor® ACE® wide pore columns, including both fully porous and solid-core particles, bonded with a variety of complementary stationary phase chemistries to optimize selectivity during method development. The use of ultrapure and highly inert base silica is emphasized for enabling the use of lower concentrations of mobile phase modifiers without compromising analyte peak shape, particularly beneficial for LC-MS applications. Then the article concludes by emphasizing the significance of reversed-phase liquid chromatography and its compatibility with mass spectrometry as a valuable tool for the separation and analysis of intact proteins and their closely related variants in biopharmaceuticals.

• Applied Chemistry for Engineering
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• v.28 no.4
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• pp.479-484
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• 2017

In this study, antioxidative activities and cellular protective effects of 70% ethanol extracts and fractions from lavender were evaluated. The scavenging activity ($FSC_{50}$) of free radical (1,1-phenyl-2-picrylhydrazyl, DPPH) was 46.6, 45.5 and $477.5{\mu}g/mL$ in the 70% ethanol extract, ethyl acetate fraction and aglycone fraction, respectively. The reactive oxygen species scavenging activities (${OSC_{50}$) of 70% ethanol extract, ethyl acetate fraction and aglycone fraction were 8.1, 3.3 and $17.6{\mu}g/mL$, respectively, and they showed lower antioxidative activity than that of using L-ascorbic acid ($1.5{\mu}g/mL$). However, the aglycone fraction showed higher photohemolysis protective effect than that of using the 70% ethanol extract and ethyl acetate fraction. At $50{\mu}M$ concentration, the cellular protective effect (${\tau}_{50}$) of 70% ethanol extract, ethyl acetate fraction and aglycone fraction from lavender was 70.6, 87.2 and 165.2 min, respectively. In particular, the lavender aglycone fraction showed 3.8 times higher cellular protective effect than that of (+)-${\alpha}$-tocopherol. The lavender fractional components including luteolin 7-O-glucuronide, vitextin, rosmarinic acid, luteolin, and apigenin were identified using TLC and LC-MS. However, the lavender aglycone fraction did not show any significant increase in flavonoids (luteolin and apigenin) compared to that of the ethyl acetate fraction. In conclusion, it is suggested that lavender may be applied as an antioxidant material in cosmetic industries.

• Journal of Applied Biological Chemistry
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• v.61 no.1
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• pp.83-91
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• 2018

Advances in bacterial and fungal genome mining uncover a plethora of cryptic secondary metabolite biosynthetic gene clusters. Guided by the genome information, targeted transcriptional derepression could be employed to determine the product of a cryptic gene cluster and to explore its biological role. Monascus spp. are food grade filamentous fungi popular in eastern Asia and several genome data belong to them are now available. We achieved transcription activation of a cryptic fungal polyketide synthase-nonribosomal peptide synthase gene Mpfus1 in Monascus purpureus ${\Delta}MpPKS5$ by inserting Aspergillus gpdA promoter at the upstream of Mpfus1 through double crossover gene replacement. The gene cluster with Mpfus1 show a high similarity to those for the biosynthesis of conjugated polyene derivatives with 2-pyrrolidone ring and the mycotoxin fusarin is the representative member of this group. The ${\Delta}MpPKS5$ is incapable of producing azaphilone pigment, providing an excellent background to identify chromogenic and UV-absorbing compounds. Activation of Mpfus1 resulted in a yellow hue on mycelia and its methanol extract exhibit a maximum absorption at 365 nm. HPLC analysis of the organic extracts indicated the presence of a variety of yellow compounds in the extract. This implies that the product of MpFus1 is metabolically or chemically unstable. LC-MS analysis guided us to predict the MpFus1 product and to propose that the Mpfus1-containing gene cluster encode the biosynthesis of a desmethyl analogue of fusarin. This study showcases the genome mining in Monascus and the possibility to unveil new biological activities embedded in it.

• Applied Chemistry for Engineering
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• v.29 no.2
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• pp.176-184
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• 2018

In the present study, we investigated the antioxidative properties, cellular protective effects and component analyses of 50% ethanol extract, ethyl acetate fraction and aglycone fraction obtained from Lysimachia christinae Hance (L. christinae Hance). In the evaluation of antioxidative properties, the free radical scavenging activities ($FSC_{50}$) of 50% ethanol extract, ethyl acetate fraction and aglycone fraction were 146.8, 22.2 and $27.2{\mu}g/mL$, respectively and total antioxidant capacities ($OSC_{50}$) were 29.3, 2.9 and $4.5{\mu}g/mL$, respectively. The ethyl acetate fraction showed the highest free radical scavenging activity and total antioxidant capacity. Also, the cellular protective effects (${\tau}_{50}$) of 50% ethanol extract, ethyl acetate fraction and aglycone fraction on $^1O_2$ induced photohemolysis of human erythrocytes were 26.9, 57.5 and 103.9 min at $5{\mu}g/mL$, respectively. In particular, ${\tau}_{50}$ of the aglycone fraction exhibited a higher cellular protective effect than that of (+)-${\alpha}$-tocopherol (37.7 min). The cell viability of the ethyl acetate fraction on the UVB-induced cell damage increased up to 90.1%. In addition, the ethyl acetate fraction ($5-25{\mu}g/mL$) showed cellular protective effects on the $H_2O_2-induced$ cell damages in a dose-dependent manner. TLC, HPLC, UV-vis spectroscopy and LC-MS were used to analyse components of the ethyl acetate fraction and the main components were quercetin, kaempferol and their glycosides. In conclusion, L. christinae Hance extract/fraction can function as antioxidants to protect the skin exposed to UV radiation and may also be used as a novel functional cosmetic material, for example, an antioxidant against skin photoaging.

• Journal of Veterinary Clinics
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• v.29 no.3
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• pp.233-236
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• 2012

Cefquinome, a fourth generation cephalosporin, has been solely used for veterinary medicine and has a broad antibacterial spectrum against gram-negatives and gram-positives being very stable to ${\beta}$-lactamases. This study was conducted to evaluate the bioequivalence of two cefquinome 2.5% products in piglets. Plasma cefquinome concentrations were analyzed by liquid chromatography-mass spectrometry (LC/MS). Mean maximum concentration ($C_{max}$) of test product ($Cequus^{(R)}$) and reference product ($Cobactan^{(R)}$) were $4.34{\pm}0.58$ and $4.22{\pm}0.47{\mu}g/mL$, and mean area under the concentration time curve ($AUC_{0{\rightarrow}{\infty}}$) values were $10.43{\pm}1.96$ and $10.25{\pm}2.98{\mu}g{\cdot}h/mL$, respectively. The 90% confidence intervals for the ratio of $C_{max}$ (0.941-1.115), and $AUC_{0{\rightarrow}{\infty}}$ (0.927-1.172) values for the test and reference products were within the acceptable bioequivalence limit of 0.80-1.25. It is concluded that two commercial cefquinome injectable solutions are bioequivalent in their extent of drug absorption in piglets.

• Food Science of Animal Resources
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• v.26 no.4
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• pp.418-425
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• 2006

This study was conducted to evaluate extraction properties of crude saponin and ginsenosides, and their effects on sensory properties of emulsified pork sausage. Non-dried ginseng root was boiled in 0 (e.g., 100% distilled water), 20, 40, 60, 80 or 100% ethanol, and powdered by a freezing dry method. Weight of dried powder for the 0% ethanol extraction was 20% of initial non-dried ginseng weight, while $20{\sim}80%$ and 100% ethanol extractions resulted in approximately 15 and 10% of their initial weights, respectively. On the other hand, crude saponin content in the dried powder was linearly increased for a higher ethanol content where 100% ethanol extraction resulted in 123.52 mg/g. LC/MS analysis of crude saponin for quantifying ginsenosides showed that Rb1, Rb2 and Rc were significantly (p<0.05) higher levels for both 80 and 100% ethanol extractions. In the case of Rg1 ginsenoside, 60, 80 and 100% ethanol extractions resulted in significantly (p<0.05) higher levels. Emulsified pork sausages containing 0, 1 or 2% ginseng extracts were smoked or non-smoked and their sensory characteristics and preference were evaluated. Smoking process significantly (p<0.05) decreased juiciness and tenderness, but the treatment significantly (p<0.05) improved flavor and consumer preference. It was particularly noticed that a 2% addition of ginseng extract prevented the adverse effects of smoking process on juiciness and tenderness while the 2% addition significantly (p<0.05) improved consumer preference. The current results implied that addition of ginseng extract in emulsified pork sausage could improve sensory quality.

• Analytical Science and Technology
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• v.27 no.5
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• pp.234-242
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• 2014

Imicyafos which is a nematicide for controlling root-knot nematodes has been registered in the Republic of Korea in 2012, and the maximum residue limits of imicyafos are set to watermelon and korean melon as each 0.05 mg/kg. Extremely reliable and sensitive analytical method is required for ensuring food safety on imicyafos residues in agricultural commodities. Imicyafos residues in samples were extracted with acetone, partitioned with hexane and dichloromethane, and then purified with florisil. The purified samples were analyzed by HPLC-UVD and confirmed with LC-MS. Linear range was between 0.1~5 mg/kg with the correlation coefficient ($r^2$) 0.99997. Average recoveries of imicyafos ranged from 77.0 to 115.4% at the spiked levels of 0.02 and 0.05 mg/kg with the relative standard deviations of 2.2~9.6%. Limit of detection and quantification were 0.005 and 0.02 mg/kg, respectively. An inter-laboratory study was conducted to validate the determination method in depth, and the results were satisfactory. All of the validation results revealed that the developed analytical method in this study is relevant for imicyafos determination in agricultural commodities and will be used as an official analytical method.

• Journal of Life Science
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• v.27 no.7
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• pp.796-804
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• 2017

This study was carried out to evaluate the antioxidant effect of methanolic extract of Houttuynia cordata (HCME) and to identify a compound having antioxidant effect. The ethyl acetate fraction of HCME showed the highest antioxidant effect in organic solvent fractions. The fraction was then separated into 12 fractions by open column chromatography. Among these fractions, the fraction 10 (Fr. 10) with the highest antioxidant activity was isolated, and its antioxidant effect was evaluated by DPPH radical scavenging activity, reducing power, TBARS, cell viability, DNA oxidation and DCF fluorescence. The Fr. 10 at a $64{\mu}g/ml$ showed 60% of inhibitory effect similar to that of vitamin C at $10{\mu}g/ml$, compared with blank group. The Fr. 10 at $64{\mu}g/ml$ showed 264% of reducing power, compared with blank group. TBARS assay showed that the Fr. 10 at $64{\mu}g/ml$ had 35.5% of inhibitory effect similar to that of vitamin E at $1,000{\mu}g/ml$, compared with blank group. The Fr. 10 above $32{\mu}g/ml$ displayed cytotoxicity. However, it was observed that the Fr. 10, above $1{\mu}g/ml$ reduced DNA damage. DCF fluorescence assay showed that the Fr. 10 inhibited oxidative stress by $H_2O_2$ in a dose dependent manner. The compound of Fr. 10 was identified to be rutin whose molecular weight is 610 by the IR and LC-MS analyses. Therefore, these results suggest that the rutin of Fr. 10 could use as a natural antioxidant for development of cosmetics and functional foods.