• Asian-Australasian Journal of Animal Sciences
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• v.32 no.7
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• pp.1052-1061
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• 2019

Objective: This study was conducted to identify duck liver-expressed antimicrobial peptide 2 (LEAP-2) and demonstrate its antimicrobial activity against various pathogens. Methods: Tissue samples were collected from 6 to 8-week-old Pekin ducks (Anas platyrhynchos domesticus), total RNA was extracted, and cDNA was synthesized. To confirm the duck LEAP-2 transcript expression levels, quantitative real-time polymerase chain reaction was conducted. Two kinds of peptides (a linear peptide and a disulfide-type peptide) were synthesized to compare the antimicrobial activity. Then, antimicrobial activity assay and fluorescence microscopic analysis were conducted to demonstrate duck LEAP-2 bactericidal activity. Results: The duck LEAP-2 peptide sequence showed high identity with those of other avian species (>85%), as well as more than 55% of identity with mammalian sequences. LEAP-2 mRNA was highly expressed in the liver with duodenum next, and then followed by lung, spleen, bursa and jejunum and was the lowest in the muscle. Both of LEAP-2 peptides efficiently killed bacteria, although the disulfide-type LEAP-2 showed more powerful bactericidal activity. Also, gram-positive bacteria was more susceptible to duck LEAP-2 than gram-negative bacteria. Using microscopy, we confirmed that LEAP-2 peptides could kill bacteria by disrupting the bacterial cell envelope. Conclusion: Duck LEAP-2 showed its antimicrobial activity against both gram-positive and gram-negative bacteria. Disulfide bonds were important for the powerful killing effect by disrupting the bacterial cell envelope. Therefore, duck LEAP-2 can be used for effective antibiotics alternatives.

• Journal of Veterinary Science
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• v.23 no.1
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• pp.1.1-1.11
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• 2022

Background: The poor bioadhesion capacity of tilmicosin resulting in treatment failure for Staphylococcus aureus small colony variants (SASCVs) mastitis. Objectives: This study aimed to increase the bioadhesion capacity of tilmicosin for the SASCVs strain and improve the antibacterial effect of tilmicosin against cow mastitis caused by the SASCVs strain. Methods: Tilmicosin-loaded chitosan oligosaccharide (COS)-sodium carboxymethyl cellulose (CMC) composite nanogels were formulated by an electrostatic interaction between COS (positive charge) and CMC (negative charge) using sodium tripolyphosphate (TPP) (ionic crosslinkers). The formation mechanism, structural characteristics, bioadhesion, and antibacterial activity of tilmicosin composite nanogels were studied systematically. Results: The optimized formulation was comprised of 50 mg/mL (COS), 32 mg/mL (CMC), and 0.25 mg/mL (TPP). The size, encapsulation efficiency, loading capacity, polydispersity index, and zeta potential of the optimized tilmicosin composite nanogels were 357.4 ± 2.6 nm, 65.4 ± 0.4%, 21.9 ± 0.4%, 0.11 ± 0.01, and -37.1 ± 0.4 mV, respectively; the sedimentation rate was one. Scanning electron microscopy showed that tilmicosin might be incorporated in nano-sized crosslinked polymeric networks. Moreover, adhesive studies suggested that tilmicosin composite nanogels could enhance the bioadhesion capacity of tilmicosin for the SASCVs strain. The inhibition zone of native tilmicosin, tilmicosin standard, and tilmicosin composite nanogels were 2.13 ± 0.07, 3.35 ± 0.11, and 1.46 ± 0.04 cm, respectively. The minimum inhibitory concentration of native tilmicosin, tilmicosin standard, and tilmicosin composite nanogels against the SASCVs strain were 2, 1, and 1 ㎍/mL, respectively. The in vitro time-killing curves showed that the tilmicosin composite nanogels increased the antibacterial activity against the SASCVs strain. Conclusions: This study provides a potential strategy for developing tilmicosin composite nanogels to treat cow mastitis caused by the SASCVs strain.

• BMB Reports
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• v.57 no.2
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• pp.71-78
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• 2024

Melanoma is one of the most aggressive skin tumors, and conventional treatment modalities are not effective in treating advanced melanoma. Although immunotherapy is an effective treatment for melanoma, it has disadvantages, such as a poor response rate and serious systemic immune-related toxic side effects. The main solution to this problem is the use of biological materials such as hydrogels to reduce these side effects and amplify the immune killing effect against tumor cells. Hydrogels have great advantages as local slow-release drug carriers, including the ability to deliver antitumor drugs directly to the tumor site, enhance the local drug concentration in tumor tissue, reduce systemic drug distribution and exhibit good degradability. Despite these advantages, there has been limited research on the application of hydrogels in melanoma treatment. Therefore, this article provides a comprehensive review of the potential application of hydrogels in melanoma immunotherapy. Hydrogels can serve as carriers for sustained drug delivery, enabling the targeted and localized delivery of drugs with minimal systemic side effects. This approach has the potential to improve the efficacy of immunotherapy for melanoma. Thus, the use of hydrogels as drug delivery vehicles for melanoma immunotherapy has great potential and warrants further exploration.

• IMMUNE NETWORK
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• v.16 no.2
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• pp.140-145
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• 2016

Ophiocordyceps sinensis is a natural fungus that has been valued as a health food and used in traditional Chinese medicine for centuries. The fungus is parasitic and colonizes insect larva. Naturally occurring O. sinensis thrives at high altitude in cold and grassy alpine meadows on the Himalayan mountain ranges. Wild Ophiocordyceps is becoming increasingly rare in its natural habitat, and its price limits its use in clinical practice. Therefore, the development of a standardized alternative is a great focus of research to allow the use of Ophiocordyceps as a medicine. To develop an alternative for wild Ophiocordyceps, a refined standardized extract, CBG-CS-2, was produced by artificial fermentation and extraction of the mycelial strain Paecilomyces hepiali CBG-CS-1, which originated from wild O. sinensis. In this study, we analyzed the in vitro immune-modulating effect of CBG-CS-2 on natural killer cells and B and T lymphocytes. CBG-CS-2 stimulated splenocyte proliferation and enhanced Th1-type cytokine expression in the mouse splenocytes. Importantly, in vitro CBG-CS-2 treatment enhanced the killing activity of the NK-92MI natural killer cell line. These results indicate that the mycelial culture extract prepared from Ophiocordyceps exhibits immune-modulating activity, as was observed in vivo and this suggests its possible use in the treatment of diseases caused by abnormal immune function.

• Korean Journal of Veterinary Research
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• v.34 no.4
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• pp.857-866
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• 1994

The present study was undertaken to examine the effects of obioactin, lonomycin A, and MDP on toxoplasmacidal activities in glycogen-induced mouse peritoneal macrophages. The killing effect of obioactin on Toxoplasma multiplication was increased significantly in proportion to its concentrations. $O_2{^-}$ generation in obioactin-treated macrophages was also increased from twofold to threefold when compared with that of untreated control. Similarly, $H_2O_2$ continued to rise in parallel with increase of the concentration of obioactin. Lonomycin A-treated macrophages also exhibited a good effect of dose-response on toxoplasmacidal activities. However, $O_2{^-}$ and $H_2O_2$ were not generated significantly in lonomycin A-treated macrophages. Macrophages treated with muramyl dipeptide (MDP) were not found to inhibit the prolifi:ration of Toxoplasma but showed the enhancement of $O_2{^-}$ and $H_2O_2$, generation. The released lysozyme levels from macrophages into cultured media were decreased tn dose-dependent fashion by in vitro treatment of obioactin, lonomycin A, and MDP. The intracellular lysozyme levels appeared to be a constant value regardless of increasing the concentrations of obioactin, lonomycin A, and MDP. Therefore, these results suggest that Toxoplasma multiplication within macrophages treated with obioactin was inhibited by the generation of $O_2{^-}$ and $H_2O_2$ and that lysozyme per se within or released from macrophages had no effect on toxoplasmacidal activity.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.88-95
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• 2013

Candida biofilms are self-organized microbial communities growing on the surfaces of host tissues and medical devices. These biofilms have been displaying increasing resistance against conventional antifungal agents. The roots of Scutellaria baicalensis have been widely used for medicinal purpose throughout East Asia. The aim of the present study was to evaluate the effect of S. baicalensis aqueous extract upon the preformed biofilms of 10 clinical C. albicans isolates, and assess the mechanism of the antibiofilm activity. Its effect on preformed biofilm was judged using an XTT reduction assay and the metabolic activity of all tested strains were reduced ($57.7{\pm}17.3$%) at MIC values. The S. baicalenis extract inhibited (1, 3)-${\beta}$-D-glucan synthase activity. The effect of S. baicalensis on the morphology of C. albicans was related to the changes in growth caused by inhibiting glucan synthesis; most cells were round and swollen, and cell walls were densely stained or ruptured. The anticandidal activity was fungicidal, and the extract also arrested C. albicans cells at $G_0/G_1$. The data suggest that S. baicalensis has multiple fatal effects on target fungi, which ultimately result in cell wall disruption and killing by inhibiting (1, 3)-${\beta}$-D-glucan synthesis. Therefore, S. baicalensis holds great promise for use in treating and eliminating biofilm-associated Candida infections.

• Korean Journal of Food Science and Technology
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• v.30 no.3
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• pp.660-664
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• 1998

Death of non-growing microorganisms in saline was studied to observe the inhibitory effect of NaCl in foods on the viability of microorganisms. When Leuconostoc mesenteroides LA10, Staphylococcus aureus B31 and Escherichia coli B34 were incubated in McIlvaine buffer with 0, 10, 20, 30% NaCl at $30^{\circ}C$, they survived best at pH 6, 5, 7, respectively. The survival of 5 lactic acid bacteria, 9 other bacteria and 2 yeasts was tested at pH 5, 6, 7 with 10% NaCl. Gram-positive bacteria survived in saline better than Gram-negative bacteria, and lactic acid bacteria and S. aureus survived better than other bacteria. The number of survivors decreased as concentrations of NaCl increased and as pH moved to acidic or alkaline side from the above-mentioned. When L. mesenteroides LA10 was incubated in saline with those materials which are known to protect microorganisms from the killing effect of NaCl, protective effect was not observed.

• Korean Journal of Materials Research
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• v.11 no.2
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• pp.120-125
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• 2001

Generally, hydroxyapatite(HAp), zeolite, carbon molecular sieve , activated carbon and alumina are used as heavy metal ions adsorption materials. Among those adsorption materials, HAp which has good positive ion-exchange ability with metal ion, and zeolite are utilized in wastewater treatment. Most of water pollutions are caused by hazardous heavy metals ions as well as bacteria in waste water. In this study, a adsorption materials (HAP and zeolite) are ion-exchanged with a well known antimicrobial metal ions, such as $Ag^+,\;Cu^{2+},\;and\;Zn^{2+}$, in order to give a adsorption of heavy metal ions and a killing effects of bacteria. The antimicrobial effects of adsorption materials are observed using by E. Coli. The results show that there is a complete antimicrobial effect in the adsorption materials with $Ag^+$ at the concentration of $1{\times}10^{-4}$cell/$m\ell$ of E. Coli until 24 hours. However, there is not good antimicrobial effects in the adsorption materials with $Cu^{2+},\;and\;Zn^{2+}$ substitution. Feng et. al. showed the denaturation effects of silver ions which induces the condensed DNA molecules and losing their replication abilities.

• Applied Chemistry for Engineering
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• v.32 no.3
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• pp.299-304
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• 2021

This study focuses on the preparation of acetaminophen (AP) imprinted functional biomaterials for a transdermal drug delivery using mung bean starch (MBS), polyvinyl alcohol (PVA), sodium benzoate (S) as a crosslinking agent, glycerol (GL) as a plasticizer, and melanin (MEL) as a photothermal agent. The prepared AP imprinted biomaterials were characterized using FE-SEM and their physical properties were evaluated. The photothermal effect and AP release property for functional biomaterials were examined with the irradiation of near infrared (NIR) laser (1.5 W/cm²). When the NIR laser was irradiated on functional biomaterials with/without the addition of MEL, the temperature of MEL added biomaterial increased from 25 ℃ to 41 ℃, whereas the biomaterial without MEL increased from 25 ℃ to 28 ℃. Results indicate that there is the photothermal effect of prepared biomaterial with the addition of MEL. Based on the results, AP release properties were evaluated using standard buffer solutions and artificial skin. It was found that AP release rates of MEL added AP loaded biomaterials were 1.2 times faster than those of MEL non-added AP loaded biomaterials when irradiating with NIR laser. We envision that the developed functional biomaterials can be utilized for an acute pain-killing treatment.

• Biomolecules & Therapeutics
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• v.30 no.6
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• pp.529-539
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• 2022

Rheumatoid arthritis (RA) is a multifactorial immune-mediated disease, the pathogenesis of which involves different cell types. T-cell activation plays an important role in RA. Therefore, inhibiting T-cell activation is one of the current therapeutic strategies. Cytotoxic T-lymphocyte antigen 4-immunoglobulin (CTLA4-Ig), also known as abatacept, reduces cytokine secretion by inhibiting T-cell activation. To achieve a homeostatic therapeutic effect, CTLA4-Ig has to be administered repeatedly over several weeks, which limits its applicability in RA treatment. To overcome this limitation, we increased the number of sialic acid-capped antennas by genetically engineering the CTLA4 region to increase the therapeutic effect of CTLA4-Ig. N-acetylglucosaminyltransferase (GnT) and α2,6-sialyltransferase (α2,6-ST) were co-overexpressed in Chinese hamster ovary (CHO) cells to generate a highly sialylated CTLA4-Ig fusion protein, named ST6. The therapeutic and immunogenic effects of ST6 and CTLA4-Ig were compared. ST6 dose-dependently decreased paw edema in a mouse model of collagen-induced arthritis and reduced cytokine levels in a co-culture cell assay in a similar manner to CTLA4-Ig. ST6- and CTLA4-Ig-induced T cell-derived cytokines were examined in CD4 T cells isolated from peripheral blood mononuclear cells after cell killing through irradiation followed by flow- and magnetic-bead-assisted separation. Interestingly, compared to CTLA4-Ig, ST6 was substantially less immunogenic and more stable and durable. Our data suggest that ST6 can serve as a novel, less immunogenic therapeutic strategy for patients with RA.