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Optimization of Streptococcus macedonicus MBF10-2 Lysate Production in Plant-based Medium by Using Response Surface Methodology

  • Andyanti, Dini;Dani, Fatin M.;Mangunwardoyo, Wibowo;Sahlan, Muhamad;Malik, Amarila
    • Microbiology and Biotechnology Letters
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    • v.47 no.2
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    • pp.220-233
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    • 2019
  • Bacterial lysates have become a common ingredient for natural health care. Lactic acid bacteria (LAB) could serve as potential candidates for lysate production: the lactic acids produced by LAB have been utilized for their moisturizing, antimicrobial, and rejuvenating effects, while other substances provide topical benefits and health effects for the skin. Our study aimed to obtain lysate from a LAB S. macedonicus MBF 10-2 through an optimized fermentation using the Response Surface Methodology. Strain MBF10-2 was cultivated in a 2L fermenter tank in de Man Rogosa and Sharpe (MRS) medium and in plant-based peptone modified MRS, i.e. Soy-peptone and Vegitone. The duration and the medium composition (dextrose and soy peptone or proteose peptone) were adjusted to obtain an optimum production of cell lysate. Central Composite Design was employed for Design Expert 7.0.0 by adjusting 3 factors: dextrose (1%, 1.5%, 2%, 2.5%, 3%), soy or proteose peptone (0.5%, 0.75%, 1%, 1.25% and 1.5%), and duration of fermentation (8, 10, 12 14, and 16 h for MRS-Soy peptone and 15, 17, 19, 21, and 23 h for MRS Vegitone). Bacteriocin-Like Inhibitor Substance activity of lysate and pH were used as indicators. The optimum condition for lysate production using MRS Soy Peptone and Vegitone are as follows: dextrose concentration 2.5%, plant-based peptone 1.25%, while optimum fermentation duration were 11.18 h (MRS Soy Peptone) and 17 h (MRS Vegitone) with a starter concentration of 10% at $OD_{600nm}$ $0.2{\pm}0.05$. However, the standard MRS medium produced better quality lysate compared to MRS plant-based peptones.

Tolerability and Effect of Early High-Dose Amino Acid Administration in Extremely Low Birth Weight Infants

  • Choi, Jin Wha;Kim, Jisook;Ahn, So Yoon;Chang, Yun Sil;Park, Won Soon;Sung, Se In
    • Neonatal Medicine
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    • v.25 no.4
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    • pp.153-160
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    • 2018
  • Purpose: The aim of this study is to examine the tolerability and effect of early high-dose amino acid administration in extremely low birth weight infants (ELBWIs). Methods: This retrospective cohort study included ELBWI (birth weight <1,000 g, n=142). Biochemical, nutritional, and neurodevelopmental data were compared between infants who received conventional low amino acid (LAA; 1.5 g/kg/day) and those who received high amino acid (HAA; 3 g/kg/day) within the first 48 hours after birth. Neurodevelopmental data included weight, height, and head circumference at discharge, 12 to 14 and 18 to 24 months of corrected age and the Korean Bayley Scale of Infant Development II (K-BSID-II) score at 18 to 24 months of corrected age. Results: The HAA group demonstrated higher peak plasma albumin ($3.0{\pm}0.4$ vs. $3.2{\pm}0.5$, P<0.05) and lower serum creatinine ($1.7{\pm}0.9$ vs. $1.4{\pm}0.8$, P<0.05) during the first 14 days than the LAA group. Full enteral feeding was achieved significantly earlier in infants in the HAA group than in infants in the LAA group ($46.2{\pm}23.0days$ vs. $34.3{\pm}21days$, P<0.01). There was no difference between the two groups in the z score changes in all growth indicators from birth to discharge and at 12 to 14 and 18 to 24 months of corrected age, as well as in the K-BSID-II score at 18 to 24 months of corrected age. Conclusion: Aggressive administration of amino acids during the first 2 days of life in ELBWI was well tolerated and correlated with earlier full enteral feeding, but did not improve growth and neurodevelopment.

Synthesis of Azo Based Disperse Dyes for Dyeing Polyester Fiber in Supercritical Carbon Dioxide (초임계 유체 염색용 아조계 분산염료 합성 및 PET 섬유에 대한 염색 특성 연구)

  • Shin, Seung-Rim;An, Kyoung-Lyong;Lee, Sunhye;Ko, Eunhee;Lee, In-Yeol;Kim, Changil;Jun, Kun
    • Textile Coloration and Finishing
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    • v.33 no.1
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    • pp.10-23
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    • 2021
  • In order to synthesize the dye suitable for supercritical carbon dioxide(ScCO2) dyeing, a series of azo based disperse dyes were prepared using various aniline derivatives as diazo components and indol derivatives as coupling components. Dyeing process in ScCO2 of the synthesized dyes was performed on PET fiber at 120℃ for 2 hrs under 250bar pressure with 0.5% o.w.f. of dye concentration. The absorption maxima varied from 400 to 580nm depending on the substituted groups in aniline derivatives and the indol derivatives. The dyes showed high molar extinction coefficients(ε) of 27,000~61,000M-1cm-1. Dyed PET fiber exhibited excellent brightness and good light, washing and perspiration(acid/alkali) fastness properties.

Stimultaneous Determination of Ephedrine Alkaloids in Ephedra sinica and Wolbigachul-tang by High Performance Liquid Chromatography (고성능 액체 크로마토그래피를 이용한 마황 및 월비가출탕 전탕액에서 에페드린류의 동시분석)

  • Song, Miyoung;Kim, Jung-Ok;Leem, HyunHee;Kim, Hojun
    • Journal of Korean Medicine for Obesity Research
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    • v.20 no.2
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    • pp.97-108
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    • 2020
  • Objectives: Ephedra sinica and Wolbigachul-tang which contains Ephedra sinica are used to treat obesity in Korean medicine. The aim of this study was to analyze the quantities of ephedrine alkaloids by high-performance liquid chromatography. Methods: The analysis was performed using a YMC-Triat C18 column with operating at 25℃, and UV detection at 210 nm. The mobile phase used a gradient flow with 0.1% H3PO4 in water and acetonitrile. Specificity, linearity, precision, accuracy, limit of detection, and limit of quantification were measured for validation anaylsis. This method was applied to analyze the quantities of ephedrine alkaloids in Ephedra sinica and Wolbigachul-tang. Results: The concentration per Ephedra sinica (gram) of ephedrine and pseudoephedrine in Ephedra sinica decoction are 4.74±0.22 mg and 2.19±0.10 mg, respectively and in Wolbigachul-tang decoction are 6.39±0.34 mg and 2.97±0.21 mg, respectively. The retention time of ephedrine was 23.6 min and that of pseudoephedrine was 25.8 min, and norephedrine and methylephedrine were not detected. Conclusions: In conclusion, analyzed the concentration of ephedrine alkaloids in Ephedra sinica and Wolbigachul-tang by the developed validation method.

Analytical characterization of O3 samples prepared for investigation of tropospheric heterogeneous reactions

  • Kim, Mihyeon;Park, Jong-Ho
    • Analytical Science and Technology
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    • v.35 no.5
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    • pp.212-217
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    • 2022
  • In this study, ozone (O3) samples were prepared for investigating the heterogeneous reactions between O3 and tropospheric aerosols and were characterized by spectroscopic methods. O3 generated from an ozone generator was purified by selective adsorption on refrigerated silica gel, followed by transfer to a sample bulb. The amount of UV light (λ = 256 nm) absorbed by O3 was measured as a function of time at two different temperatures (room temperature and 50 ℃) and under different irradiation conditions. A correlation plot of 1/[O3] versus time showed that O3 decomposition follows the 2nd order reaction rate under a steady-state approximation. The initial concentration of O3, observed rate constants (kobs), and the half-life of O3 in the sample stored at room temperature were determined to be 2.74 [±0.14] × 1016 molecules·cm-3, 4.47 [±0.64] × 10-23 molecules-1·cm3·s-1, and 9.5 [±1.4] days, respectively. The evaluation of O3 stability under various conditions indicated that special care should be taken to prevent the exposure of the O3 samples to hightemperature environment and/or UV radiation. This study established a protocol for the preparation of highly purified O3 samples and confirmed that the O3 samples can be stored for a day after preparation for further experiments.

Novel Antibacterial, Cytotoxic and Catalytic Activities of Silver Nanoparticles Synthesized from Acidophilic Actinobacterial SL19 with Evidence for Protein as Coating Biomolecule

  • Wypij, Magdalena;Ostrowski, Maciej;Piska, Kamil;Wojcik-Pszczola, Katarzyna;Pekala, Elzbieta;Rai, Mahendra;Golinska, Patrycja
    • Journal of Microbiology and Biotechnology
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    • v.32 no.9
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    • pp.1195-1208
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    • 2022
  • Silver nanoparticles (AgNPs) have potential applications in medicine, photocatalysis, agriculture, and cosmetic fields due to their unique physicochemical properties and strong antimicrobial activity. Here, AgNPs were synthesized using actinobacterial SL19 strain, isolated from acidic forest soil in Poland, and confirmed by UV-vis and FTIR spectroscopy, TEM, and zeta potential analysis. The AgNPs were polydispersed, stable, spherical, and small, with an average size of 23 nm. The FTIR study revealed the presence of bonds characteristic of proteins that cover nanoparticles. These proteins were then studied by using liquid chromatography with tandem mass spectrometry (LC-MS/MS) and identified with the highest similarity to hypothetical protein and porin with molecular masses equal to 41 and 38 kDa, respectively. Our AgNPs exhibited remarkable antibacterial activity against Escherichia coli and Pseudomonas aeruginosa. The combined, synergistic action of these synthesized AgNPs with commercial antibiotics (ampicillin, kanamycin, streptomycin, and tetracycline) enabled dose reductions in both components and increased their antimicrobial efficacy, especially in the case of streptomycin and tetracycline. Furthermore, the in vitro activity of the AgNPs on human cancer cell lines (MCF-7, A375, A549, and HepG2) showed cancer-specific sensitivity, while the genotoxic activity was evaluated by Ames assay, which revealed a lack of mutagenicity on the part of nanoparticles in Salmonella Typhimurium TA98 strain. We also studied the impact of the AgNPs on the catalytic and photocatalytic degradation of methyl orange (MO). The decomposition of MO was observed by a decrease in intensity of absorbance within time. The results of our study proved the easy, fast, and efficient synthesis of AgNPs using acidophilic actinomycete SL19 strain and demonstrated the remarkable potential of these AgNPs as anticancer and antibacterial agents. However, the properties and activity of such particles can vary by biosynthesized batch.

Endoscopic Laser Irradiation Condition of Urethra in Tubular Structure (관형 요도 조직 대상 내시경적 레이저 조사 조건 연구)

  • Hwarang, Shin;Seonghee, Lim;Yeachan, Lee;Hyun Wook, Kang
    • Journal of Biomedical Engineering Research
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    • v.44 no.1
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    • pp.85-91
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    • 2023
  • Stress urinary incontinence (SUI) occurs when abdominal pressure increases, such as sneezing, exercising, and laughing. Surgical and non-surgical treatments are the common methods of SUI treatment; however, the conventional treatments still require continuous and invasive treatment. Laser have been used to treat SUI, but excessive temperature increase often causes thermal burn on urethra tissue. Therefore, the optimal conditions must be considered to minimize the thermal damage for the laser treatment. The current study investigated the feasibility of the laser irradiation condition for SUI treatment using non-ablative 980 nm laser from a safety perspective through numerical simulations. COMSOL Multiphysics was used to analyze the numerical simulation model. The Pennes bioheat equation with the Beer's law was used to confirm spatio-temporal temperature distributions, and Arrhenius equation defined the thermal damage caused by the laser-induced heat. Ex vivo porcine urethral tissue was tested to validate the extent of both temperature distribution and thermal damage. The temperature distribution was symmetrical and uniformly observed in the urethra tissue. A muscle layer had a higher temperature (28.3 ℃) than mucosal (23.4 ℃) and submucosal layers (25.5 ℃). MT staining revealed no heat-induced collagen and muscle damage. Both control and treated groups showed the equivalent thickness and area of the urethral mucosal layer. Therefore, the proposed numerical simulation can predict the appropriate irradiation condition (20 W for 15 s) for the SUI treatment with minimal temperature-induced tissue.

Antibacterial activity of enrofloxacin loaded gelatin-sodium alginate composite nanogels against intracellular Staphylococcus aureus small colony variants

  • Luo, Wanhe;Liu, Jinhuan;Algharib, Samah Attia;Chen, Wei
    • Journal of Veterinary Science
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    • v.23 no.3
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    • pp.48.1-48.12
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    • 2022
  • Background: The poor intracellular concentration of enrofloxacin might lead to treatment failure of cow mastitis caused by Staphylococcus aureus small colony variants (SASCVs). Objectives: In this study, enrofloxacin composite nanogels were developed to increase the intracellular therapeutic drug concentrations and enhance the efficacy of enrofloxacin against cow mastitis caused by intracellular SASCVs. Methods: Enrofloxacin composite nanogels were formulated by an electrostatic interaction between gelatin (positive charge) and sodium alginate (SA; negative charge) with the help of CaCl2 (ionic crosslinkers) and optimized by a single factor test using the particle diameter, zeta potential (ZP), polydispersity index (PDI), loading capacity (LC), and encapsulation efficiency (EE) as indexes. The formation mechanism, structural characteristics, bioadhesion ability, cellular uptake, and the antibacterial activity of the enrofloxacin composite nanogels against intracellular SASCVs strain were studied systematically. Results: The optimized formulation was comprised of 10 mg/mL (gelatin), 5 mg/mL (SA), and 0.25 mg/mL (CaCl2). The size, LC, EE, PDI, and ZP of the optimized enrofloxacin composite nanogels were 323.2 ± 4.3 nm, 15.4% ± 0.2%, 69.6% ± 1.3%, 0.11 ± 0.02, and -34.4 ± 0.8 mV, respectively. Transmission electron microscopy showed that the enrofloxacin composite nanogels were spherical with a smooth surface and good particle size distributions. In addition, the enrofloxacin composite nanogels could enhance the bioadhesion capacity of enrofloxacin for the SASCVs strain by adhesive studies. The minimum inhibitory concentration, minimum bactericidal concentration, minimum biofilm inhibitory concentration, and minimum biofilm eradication concentration were 2, 4, 4, and 8 ㎍/mL, respectively. The killing rate curve had a concentration-dependent bactericidal effect as increasing drug concentrations induced swifter and more radical killing effects. Conclusions: This study provides a good tendency for developing enrofloxacin composite nanogels for treating cow mastitis caused by intracellular SASCVs and other intracellular bacterial infections.

Antibacterial activity of florfenicol composite nanogels against Staphylococcus aureus small colony variants

  • Liu, Jinhuan;Ju, Mujie;Wu, Yifei;Leng, Nannan;Algharib, Samah Attia;Luo, Wanhe
    • Journal of Veterinary Science
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    • v.23 no.5
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    • pp.78.1-78.13
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    • 2022
  • Background: Florfenicol might be ineffective for treating Staphylococcus aureus small colony variants (SCVs) mastitis. Objectives: In this study, florfenicol-loaded chitosan (CS)-sodium tripolyphosphate (TPP) composite nanogels were prepared to allow targeted delivery to SCV infected sites. Methods: The formulation screening, the characteristics, in vitro release, antibacterial activity, therapeutic efficacy, and biosafety of the florfenicol composite nanogels were studied. Results: The optimized formulation was obtained when the CS and TPP were 10 and 5 mg/mL, respectively. The encapsulation efficiency, loading capacity, size, polydispersity index, and zeta potential of the optimized florfenicol composite nanogels were 87.3% ± 2.7%, 5.8% ± 1.4%, 280.3 ± 1.5 nm, 0.15 ± 0.03, and 36.3 ± 1.4 mv, respectively. Optical and scanning electron microscopy showed that spherical particles with a relatively uniform distribution and drugs might be incorporated in cross-linked polymeric networks. The in vitro release study showed that the florfenicol composite nanogels exhibited a biphasic pattern with the sustained release of 72.2% ± 1.8% at 48 h in pH 5.5 phosphate-buffered saline. The minimal inhibitory concentrations of commercial florfenicol solution and florfenicol composite nanogels against SCVs were 1 and 0.25 ㎍/mL, respectively. The time-killing curves and live-dead bacterial staining showed that the florfenicol composite nanogels were concentration-dependent. Furthermore, the florfenicol composite nanogels displayed good therapeutic efficacy against SCVs mastitis. Biological safety studies showed that the florfenicol composite nanogels might be a biocompatible preparation because of their non-toxic effects on the renal tissue and liver. Conclusions: Florfenicol composite nanogels might improve the treatment of SCV infections.

Classification of Red Wines by Near Infrared Transflectance Spectroscopy

  • W.Guggenbichler;Huck, C.W.;M.Popp;G.K.Bonn
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.1516-1516
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    • 2001
  • During the recent years, wine analysis has played an increasing role due the health benefits of phenolic ingredients in red wine [1]. On the other hand there is the need to be able to distinguish between different wine varieties. Consumers want to know if a wine is an adulterated one or if it is based on the pure grape. Producers need to certificate their wines in order to ensure compliance with legal regulations. Up to now, the attempts to investigate the origin of wines were based on high-performance liquid chromatography (HPLC), gas chromatography (GC) and pyrolysis mass spectrometry (PMS) [l,2,3]. These methods need sample pretreatment, long analysis times and therefore lack of high sample throughput. In contradiction to these techniques using near infrared spectroscopy (NIRS), no sample pretreatment is necessary and the analysis time for one sample is only about 10 seconds. Hence, a near infrared spectroscopic method is presented that allows a fast classification of wine varieties in bottled red wines. For this, the spectra of 50 bottles of Cabernet Sauvignon, Lagrein and Sangiovese (Chianti) were recorded without any sample pretreatment over a wavelength range from 1000 to 2500 nm with a resolution of 12 cm$\^$-1/. 10 scans were used for an average spectrum. In order to yield best reproducibility, wines were thermostated at 23$^{\circ}C$ and a optical layer thickness of 3 mm was used. All recorded spectra were partitioned into a calibration and validation set (70% and 30%). Finally, a 3d scatter plot of the different investigated varieties allowed to distinguish between Cabernet Sauvignon, Lagrein and Sangiovese (Chianti). Considering the short analysis times this NRS-method will be an interesting tool for the quality control of wine verification and also for experienced sommeliers.

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