• Food Science of Animal Resources
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• v.32 no.6
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• pp.706-712
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• 2012

The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 ${\mu}g/mL$, having ca. $10^8$ coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of $10^5$ CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.6
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• pp.942-949
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• 2010

Morus alba L., a kind of Oriental medicinal herbs, has been traditionally used to treat pulmonary asthma and congestion. According to recent studies, extracts of M. alba L. have showed anti-inflammatory, anti-oxidant, anti-tumor and hypoglycemic effects. However, the molecular mechanisms on how it acts as a death-inducer in cancer cells have not been fully understood. In this study, we investigated the cell death effects of methylene chloride extracts of M. alba L. (MEMA) in A549 human lung carcinoma cells. It was shown that MEMA induced the apoptotic cell death proved by increased sub-G1 phase cell population, apoptotic body formation and chromatin condensation. MEMA treatment induced the expression of death receptor-related proteins such as death receptor (DR) 4, DR5, Fas and FasL, which further triggered the activation of caspase-8 and the cleavage of Bid in a concentration-dependent manner. However, MEMA reduced anti-apoptotic Bcl-2 and Bcl-xL expression which contributed to the loss of mitochondrial membrane potential (MMP), and the activations of caspase-9 and caspase-3. Meanwhile, the morphological study indicated a characteristic finding of autophagy, such as the formation of autophagosomes in MEMA-treated cells. Furthermore, markers of autophagy, namely, the increased MDC-positive cells, conversion of microtubule-associated protein light chain 3 (LC3)-I to LC3-II and increased beclin-1 accumulation, were observed. Taken together, these findings demonstrated that MEMA triggered both autophagy and apoptosis in A549 cancer cells. They might suggest that M. alba L. could be a prospective clinical application to treat human lung cancers.

• Journal of Veterinary Clinics
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• v.37 no.2
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• pp.106-108
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• 2020

An eight-month-old, outdoor, intact male English Pointer dog weighing 23.5 kg presented to the hospital with signs of hematochezia, soft stools, and weight-loss. There were no remarkable findings on physical examination, complete blood count, serum biochemistry, electrolyte and gas analysis, and radiography. The serologic and Polymerase Chain Reaction (PCR) tests for canine parvovirus were negative. A fecal smear examination showed rod-shaped, sporeforming bacteria. Additionally, a fecal flotation test showed ova of Ancylostoma spp. The size of ova was 60 × 40 ㎛, and it was identified as Ancylostoma caninum using light microscopy. The PCR test indicated a Clostridial perfringens infection and the presence of C. perfringens alpha toxin. The diagnosis given was C. perfringens enterotoxicosis with ancylostomiasis. Treatment included antibiotics (metronidazole, trimethoprim-sulfamethoxazole) and anthelmintics (afoxolaner, milbemycin oxime). After two weeks, the clostridial infection resolved, but ancylostomiasis persisted for six weeks. The anthelmintic was changed to Drontal^â plus (praziquantel/pyrantel pamoate/febantel). After four weeks, there were no remarkable findings in the fecal samples, but the patient still presented with watery stools and hematochezia. Survey of abdominal ultrasound had performed, and a target-like sign with multiple rings was seen in the cecocolic region. The patient was diagnosed with A. caninum-induced cecocolic intussusception from the history and clinical signs. After a surgery, he recovered fully. This is the first clinical case report of Ancylostoma caninum parasitizing from the small intestine and causing an intussusception in the large intestine.

• Molecules and Cells
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• v.23 no.2
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• pp.175-181
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• 2007

The present study was undertaken to determine whether $SM22{\alpha}$ participates in the regulation of vascular smooth muscle contractility using $SM22{\alpha}$ knockout mice and, if so, to investigate the mechanisms involved. Aortic ring preparations were mounted and equilibrated in organ baths for 60 min before observing contractile responses to 50 mM KCl, and then exposed to contractile agents such as phenylephrine and phorbol ester. Measurement of isometric contractions using a computerized data acquisition system was combined with molecular or cellular experiments. Interestingly, the aortas from $SM22{\alpha}$-deficient mice ($SM22^{-/-LacZ}$) displayed an almost three-fold increase in the level of $SM22{\beta}$ protein compared to wild-type mice, but no change in the levels of caldesmon, actin, desmin or calponin. $Ca^{2+}$-independent contraction in response to phenylephrine or phorbol ester was significantly decreased in the $SM22{\alpha}$-deficient mice, whereas in the presence of $Ca^{2+}$ neither contraction nor subcellular translocation of myosin light chain kinase (MLCK) in response to phenylephrine or 50 mM KCl was significantly affected. A decrease in phosphorylation of extracellular signal regulated kinase (ERK) 1/2 was observed in the $SM22{\alpha}$-deficient mice and this may be related to the decreased vascular contractility. Taken together, this study provides evidence for a pivotal role of $SM22{\alpha}$ in the regulation of $Ca^{2+}$-independent vascular contractility.

• Molecules and Cells
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• v.39 no.9
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• pp.669-679
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• 2016

N-acetyl-D-glucosamine kinase (GlcNAc kinase or NAGK) primarily catalyzes phosphoryl transfer to GlcNAc during amino sugar metabolism. Recently, it was shown NAGK interacts with dynein light chain roadblock type 1 (DYNLRB1) and upregulates axo-dendritic growth, which is an enzyme activity-independent, non-canonical structural role. The authors examined the distributions of NAGK and NAGK-dynein complexes during the cell cycle in HEK293T cells. NAGK was expressed throughout different stages of cell division and immunocytochemistry (ICC) showed NAGK was localized at nuclear envelope, spindle microtubules (MTs), and kinetochores (KTs). A proximity ligation assay (PLA) for NAGK and DYNLRB1 revealed NAGK-dynein complex on nuclear envelopes in prophase cells and on chromosomes in metaphase cells. NAGK-DYNLRB1 PLA followed by Lis1/NudE1 immunostaining showed NAGK-dynein complexes were colocalized with Lis1 and NudE1 signals, and PLA for NAGK-Lis1 showed similar signal patterns, suggesting a functional link between NAGK and dynein-Lis1 complex. Subsequently, NAGK-dynein complexes were found in KTs and on nuclear membranes where KTs were marked with CENP-B ICC and nuclear membrane with lamin ICC. Furthermore, knockdown of NAGK by small hairpin (sh) RNA was found to delay cell division. These results indicate that the NAGK-dynein interaction with the involvements of Lis1 and NudE1 plays an important role in prophase nuclear envelope breakdown (NEB) and metaphase MT-KT attachment during eukaryotic cell division.

• Korean Journal of Microbiology
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• v.36 no.1
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• pp.74-78
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• 2000

Vibrio, Photobacterium, Alteromonas and Xenorhabdus species are capable of emitting light, called bioluminescence. They exist in marine, freshwater and terrestrial environments. Bacterial bioluminescent reaction is that reduced riboflavin phosphates and a long-chain aldehyde are oxidized in the presence of molecular oxygen and enzyme luciferase. This experiment aims to develop the proper culture media and to optimize the storage condition for the recovery of bioluminescent activity in Photobacterium phosphoreum. The Luria broth (LB) medium was modified for cultivation of Photobacterium phophoreum, called as modified LB(mLB) medium. The mLB medium is LB fortified with 3% glycerol and 1.5% NaCl. In mLB medium. bacterial growth and bioluminescent activity are 25% higher than those in a Nutrient broth medium. When the cell stocks were stored at $-20^{\circ}C$, $-70^{\circ}C$ and LN2 for 3 months, cell growth and bioluminescent activity of culture after stored at $-20^{\circ}C$ were better than those of other treatments. The highest bioluminescent activity obtained at the late exponential phase in all treatments. When the cell stock was freeze-dried with 5% adonitol as a cryoprotectant, the recovery of cell was better than those of control and freeze-dried cell stock without addition of cryoprotectant.

• Journal of Periodontal and Implant Science
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• v.44 no.6
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• pp.280-287
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• 2014

Purpose: The purpose of this randomized single-blind controlled trial was to elucidate the clinical and antimicrobial effects of daily phototherapy (PT) as an adjunct to scaling and root planing (SRP) in patients with chronic periodontitis. Methods: The study was conducted from December 2013 to May 2014 at Ewha Womans University Mokdong Hospital, Seoul, Korea. Forty-one patients with mild to moderate chronic periodontitis were randomly divided into two therapeutic groups in a 1:1 ratio: SRP+PT and SRP (control) groups. All participants underwent full-mouth SRP. PT was performed thrice a day for a month by using electric toothbrushes with embedded light-emitting diodes. Plaque index, gingival index, probing pocket depth (PPD), clinical attachment level (CAL), and bleeding on probing were assessed before (baseline) and four weeks after (follow-up) the treatment. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum, Parvimonas micra, Campylobacter rectus, Eikenella corrodens, Streptococcus mutans, and Streptococcus sobrinus levels were detected by a real-time polymerase chain reaction at the same points in time. Results: The clinical parameters improved in both the groups. At the follow-up assessment, PPD was significantly decreased in the SRP+PT group (P=0.00). Further, PPD and CAL showed significantly greater changes in the SRP+PT group than in the SRP group (PPD, P=0.03; CAL, P=0.04). P. gingivalis and T. forsythia levels decreased in this group, but no significant intergroup differences were noted. Conclusions: Adjunctive PT seems to have clinical benefits, but evidence of its antimicrobial effects is not sufficient. Long-term studies are necessary to develop the most effective PT protocol and compare the effectiveness of PT with and without exogenous photosensitizers.

• Korean Chemical Engineering Research
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• v.58 no.1
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• pp.122-126
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• 2020

In this study, we produced the micropatterned channel system using polydimethylsiloxane (PDMS) and micromolding in capillaries (MIMIC) technology and evaluated cellular polarity signals through high-resolved imaging at the single-cell level. In cells treated with platelet-derived growth factor (PDGF), three types of key signals in cell migration; phosphoinositide 3-kinase (PI3 K), Rac, and Actin, were strongly activated in the front area compared to the rear region, whereas myosin light chain (MLC) showed no notable activity in the front and rear areas. Our results will, therefore, provide important information and methodology for studying the correlation between cell polarity signals and cell migration under the newly defined microenvironment.

• The Korea Journal of Herbology
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• v.31 no.5
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• pp.41-46
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• 2016

Objectives : The Skin is composed of multiple layers, including the epidermis, dermis, and hypodermis. It provides a vital barrier structure that protects vertebrates from external environmental antigens, solvents, ultraviolet light, microorganisms, toxins, and weather conditions. Although several biological effects of Mori Follium have been reported, beneficial effects of Mori Follium in skin health remain unclear. In this study, we prepared water extract of Mori Follium (MLE) and evaluated the effects on melanin accumulation and expression levels of skin fibril-related proteins.Methods : The cytotoxicities of MLE in B16F10 melanoma and human skin fibroblasts (HSF) were examined by MTT assay. Inhibitory effect of MLE on the α-MSH- and IBMX-induced melanosis in B16F10 melanoma was examined. The expression levels of fibronectin, collagen 1α2, and CCN2 in MLE-treated HSF were analyzed by reverse transcription-polymer chain reaction (RT-PCR) and western blotting.Results : The MLE treatment for 24 h did not affect to the B16F10 and HSF at concentrations of 1, 10, 50, 100, 200, 400 and 800 ㎍/ml. The MLE treatment for 72 h significantly and dose dependently suppressed melanin accumulation in B16F10 melanoma. In addition, the MLE treatment up-regulated expression levels of skin fibril-related genes such as fibronectin, collagen 1α2, and CCN2 in HSF. Our western blot analysis revealed MLE-induced up-regulation of skin fibril-related genes required the activation of CCN2 protein.Conclusions : In conclusion, these findings suggest that the MLE could be used in development of cosmetic natural material of maintaining healthy skin.

• Animal cells and systems
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• v.5 no.3
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• pp.253-262
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• 2001

Ligand-receptor clustering event is the most important step in leukocyte adhesion and spreading on endothelial cells. Intercellular adhesion molecule-1 (ICAM-1) has been shown to enhance leukocyte adhesion, but the molecular event during the process of adhesion is unclear. To visualize the dynamics of ICAM-1 movement during adhesion, we have engineered stable Chinese hamster ovary cell lines expressing ICAM-1 fused to a green fluorescent protein (IC1_GFP/CHO) and examined them under the fluorescence microscopy. The transfection of IC1_GFP alone in these cells was sufficient to support the adhesion of K562 cells that express $\alpha$L$\beta$2 (LFA-1) integrin stimulated by CBR LFA-1/2 mAb. This phenomenon was mediated by ICAM-1-LFA-1 interactions, as an mAb that specifically inhibits ICAM-1-LFA-1 interaction (RRl/l) completely abolished the adhesion of LFA-1＊ cells to IC1_ GFP/CHO cells. We found that the characteristic of adhesion was followed almost immediately (~10 min) by the rapid accumulation of ICAM-1 on CHO cells at a tight interface between the two cells. Interestingly, ICI_GFP/CHO cells projected plasma membrane and encircled approximately half surface of LFA-1+ cells, as defined by confocal microscopy. This unusual phenomenon was also confirmed on HUVEC after adhesion of LFA-1＊ cells. Neither cytochalasin D nor 2,3-butanedione 2-monoxime an inhibitor of myosin light chain kinase blocked LFA-1-mediated ICAM-1 clustering, indicating that actin cytoskeleton and myosin-dependent contractility are not necessary for ICAM-1 clustering. Taken together, we suggest that leukocyte adhesion to endothelial cells induces specialized form of ICAM-1 clustering that is distinct from immunological synapse mediated by T cell interaction with antigen presenting cells.