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Multilocus sequence type-dependent activity of human and animal cathelicidins against community-, hospital-, and livestock-associated methicillin-resistant Staphylococcus aureus isolates

  • Sun Do, Kim;Geun-Bae, Kim;Gi Yong, Lee;Soo-Jin, Yang
    • Journal of Animal Science and Technology
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    • v.64 no.3
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    • pp.515-530
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    • 2022
  • Sequence type (ST) 5 methicillin-resistant Staphylococcus aureus (MRSA) with staphylococcal cassette chromosome mec (SCCmec) type II (ST5-MRSA-II) and ST72-MRSA-IV represent the most significant genotypes for healthcare- (HA) and community-associated (CA) MRSA in Korea, respectively. In addition to the human-type MRSA strains, the prevalence of livestock-associated (LA) MRSA clonal lineages, such as ST541 and ST398 LA-MRSA-V in pigs and ST692 LA-MRSA-V and ST188 LA-MRSA-IV in chickens, has recently been found. In this study, clonotype-specific resistance profiles to cathelicidins derived from humans (LL-37), pigs (PMAP-36), and chickens (CATH-2) were examined using six different ST groups of MRSA strains: ST5 HA-MRSA-II, ST72 CA-MRSA-IV, ST398 LA-MRSA-V, ST541 LA-MRSA-V, ST188 LA-MRSA-IV, and ST692 LA-MRSA-V. Phenotypic characteristics often involved in cathelicidin resistance, such as net surface positive charge, carotenoid production, and hydrogen peroxide susceptibility were also determined in the MRSA strains. Human- and animal-type MRSA strains exhibited clonotype-specific resistance profiles to LL-37, PMAP-36, or CATH-2, indicating the potential role of cathelicidin resistance in the adaptation and colonization of human and animal hosts. The ST5 HA-MRSA isolates showed enhanced resistance to all three cathelicidins and hydrogen peroxide than ST72 CA-MRSA isolates by implementing increased surface positive charge and carotenoid production. In contrast, LA-MRSA strains employed mechanisms independent of surface charge regulation and carotenoid production for cathelicidin resistance. These results suggest that human- and livestock-derived MRSA strains use different strategies to counteract the bactericidal action of cathelicidins during the colonization of their respective host species.

Species Profiles and Antimicrobial Resistance of Non-aureus Staphylococci Isolated from Healthy Broilers, Farm Environments, and Farm Workers

  • Ji Heon Park;Gi Yong Lee;Ji Hyun Lim;Geun-Bae Kim;Kun Taek Park;Soo-Jin Yang
    • Food Science of Animal Resources
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    • v.43 no.5
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    • pp.792-804
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    • 2023
  • Non-aureus staphylococci (NAS), particularly antimicrobial-resistant NAS, have a substantial impact on human and animal health. In the current study, we investigated (1) the species profiles of NAS isolates collected from healthy broilers, farm environments, and farm workers in Korea, (2) the occurrence of antimicrobial-resistant NAS isolates, especially methicillin resistance, and (3) the genetic factors involved in the methicillin and fluoroquinolone resistance. In total, 216 NAS isolates of 16 different species were collected from healthy broilers (n=178), broiler farm environments (n=18), and farm workers (n=20) of 20 different broiler farms. The two most dominant broiler-associated NAS species were Staphylococcus agnetis (23.6%) and Staphylococcus xylosus (22.9%). Six NAS isolates were mecA-positive carrying staphylococcal cassette chromosome mec (SCCmec) II (n=1), SCCmec IV (n=1), SCCmec V (n=2), or nontypeable SCCmec element (n=2). While two mecA-positive Staphylococcus epidermidis isolates from farm workers had SCCmec II and IV, a mecA-positive S. epidermidis isolate from broiler and a Staphylococcus haemolyticus isolate farm environment carried SCCmec V. The occurrence of multidrug resistance was observed in 48.1% (104/216 isolates) of NAS isolates with high resistance rates to β-lactams (>40%) and fusidic acid (59.7%). Fluoroquinolone resistance was confirmed in 59 NAS isolates (27.3%), and diverse mutations in the quinolone resistance determining regions of gyrA, gyrB, parC, and parE were identified. These findings suggest that NAS in broiler farms may have a potential role in the acquisition, amplification, and transmission of antimicrobial resistance.

Molecular characteristics of Escherichia coli from bulk tank milk in Korea

  • Yoon, Sunghyun;Lee, Young Ju
    • Journal of Veterinary Science
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    • v.23 no.1
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    • pp.9.1-9.11
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    • 2022
  • Background: Escherichia coli, which causes subclinical or clinical mastitis in cattle, is responsible for transmitting antimicrobial resistance via human consumption of raw milk or raw milk products. Objectives: The objective of this study was to investigate the molecular characteristics of 183 E. coli from bulk tank milk of five different dairy factories in Korea. Methods: The molecular characteristics of E. coli such as serogroup, virulence, antimicrobial resistance, and integron genes were detected using polymerase chain reaction and antimicrobial susceptibility were tested using the disk diffusion test. Results: In the distribution of phylogenetic groups, group D was the most prevalent (59.6%) and followed by group B1 (25.1%). The most predominant serogroup was O173 (15.3%), and a total of 46 different serotypes were detected. The virulence gene found most often was fimH (73.2%), and stx1, fimH, incC, fyuA, and iutA genes were significantly higher in isolates of phylogenetic group B1 compared to phylogenetic groups A, B2, and D (p < 0.05). Among 64 E. coli isolates that showed resistance to at least one antimicrobial, the highest resistance rate was observed for tetracyclines (37.5%). All 18 integron-positive E. coli carried the integron class I (int1) gene, and three different gene cassette arrangements, dfrA12+aadA2 (2 isolates), aac(6')-Ib3+aac(6')-Ib-cr+aadA4 (2 isolates), and dfrA17+aadA5 (1 isolate) were detected. Conclusions: These data suggest that the E. coli from bulk tank milk can be an indicator for dissemination of antimicrobial resistance and virulence factors via cross-contamination.

Chemoquiescence with Molecular Targeted Ablation of Cancer Stem Cells in Gastrointestinal Cancers

  • Jong-Min Park;Young-Min Han;Migyeong Jeong;Eun Jin Go;Napapan Kangwan;Woo Sung Kim;Ki Baik Hahm
    • Journal of Digestive Cancer Research
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    • v.4 no.1
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    • pp.1-9
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    • 2016
  • The abundance of multi-drug resistance ATPase binding cassette and deranged self-renewal pathways shown in cancer stem cells (CSCs) played a crucial role in tumorigenesis, tumor resistance, tumor recurrence, and tumor metastasis. Therefore, elucidation of CSCs biology can improve diagnosis, enable targeted treatment, and guide the follow up of GI cancer patients. In order to achieve chemoquiescence, seizing cancer through complete ablation of CSCs, CSCs are rational targets for the design of interventions that will enhance responsiveness to traditional therapeutic strategies and contribute in the prevention of local recurrence as well as metastasis. However, current cancer treatment strategies fail to either detect or differentiate the CSCs from their non-tumorigenic progenies mostly due to the absence of specific biomarkers and potent agents to kill CSCs. Recent advances in knowledge of CSCs enable to produce several candidates to ablate CSCs in gastrointestinal (GI) cancers, especially cancers originated from inflammation-driven mutagenesis such as Barrett's esophagus (BE), Helicobacter pylori-associated gastric cancer, and colitis-associated cancer (CAC). Our research teams elucidated through revisiting old drugs that proton pump inhibitor (PPI) and potassium competitive acid blocker (p-CAB) beyond authentic acid suppression, chloroquine for autophage inhibition, sonic hedgehog (SHH) inhibitors, and Wnt/β-catenin/NOTCH inhibitor can ablate CSCs specifically and efficiently. Furthermore, nanoformulations of these molecules could provide an additional advantage for more selective targeting of the pathways existing in CSCs just like current molecular targeted therapeutics and sustained action, while normal stem cells intact. In this review article, the novel approach specifically to ablate CSCs existing in GI cancers will be introduced with the introduction of explored mode of action.

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Glucosamine increases macrophage lipid accumulation by regulating the mammalian target of rapamycin signaling pathway

  • Sang-Min Kim;Dong Yeol Kim;Jiwon Park;Young-Ah Moon;Inn-Oc Han
    • BMB Reports
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    • v.57 no.2
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    • pp.92-97
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    • 2024
  • Elevated blood glucose is associated with an increased risk of atherosclerosis. Data from the current study showed that glucosamine (GlcN), a normal glucose metabolite of the hexosamine biosynthetic pathway (HBP), promoted lipid accumulation in RAW264.7 macrophage cells. Oleic acid- and lipopolysaccharide (LPS)-induced lipid accumulation was further enhanced by GlcN in RAW264.7 cells, although there was no a significant change in the rate of fatty acid uptake. GlcN increased acetyl CoA carboxylase (ACC), fatty acid synthase (FAS), scavenger receptor class A, liver X receptor, and sterol regulatory element-binding protein-1c (SREBP-1c) mRNA expression, and; conversely, suppressed ATP-binding cassette transporter A1 (ABCA-1) and ABCG-1 expression. Additionally, GlcN promoted O-GlcNAcylation of nuclear SREBP-1 but did not affect its DNA binding activity. GlcN stimulated phosphorylation of mammalian target of rapamycin (mTOR) and S6 kinase. Rapamycin, a mTOR-specific inhibitor, suppressed GlcN-induced lipid accumulation in RAW264.7 cells. The GlcN-mediated increase in ACC and FAS mRNA was suppressed, while the decrease in ABCA-1 and ABCG-1 by GlcN was not significantly altered by rapamycin. Together, our results highlight the importance of the mTOR signaling pathway in GlcN-induced macrophage lipid accumulation and further support a potential link between mTOR and HBP signaling in lipogenesis.

Ellagic acid, a functional food component, ameliorates functionality of reverse cholesterol transport in murine model of atherosclerosis

  • Sin-Hye Park;Min-Kyung Kang;Dong Yeon Kim;Soon Sung Lim;Il-Jun Kang;Young-Hee Kang
    • Nutrition Research and Practice
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    • v.18 no.2
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    • pp.194-209
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    • 2024
  • BACKGROUND/OBJECTIVES: High levels of plasma low-density lipoprotein (LDL) cholesterol are an important determinant of atherosclerotic lesion formation. The disruption of cholesterol efflux or reverse cholesterol transport (RCT) in peripheral tissues and macrophages may promote atherogenesis. The aim of the current study was to examine whether bioactive ellagic acid, a functional food component, improved RCT functionality and high-density lipoprotein (HDL) function in diet-induced atherogenesis of apolipoproteins E (apoE) knockout (KO) mice. MATERIALS/METHODS: Wild type mice and apoE KO mice were fed a high-cholesterol Paigen diet for 10 weeks to induce hypercholesterolemia and atherosclerosis, and concomitantly received 10 mg/kg ellagic acid via gavage. RESULTS: Supplying ellagic acid enhanced induction of apoE and ATP-binding cassette (ABC) transporter G1 in oxidized LDL-exposed macrophages, facilitating cholesterol efflux associated with RCT. Oral administration of ellagic acid to apoE KO mice fed on Paigen diet improved hypercholesterolemia with reduced atherogenic index. This compound enhanced the expression of ABC transporters in peritoneal macrophages isolated from apoE KO mice fed on Paigen diet, indicating increased cholesterol efflux. Plasma levels of cholesterol ester transport protein and phospholipid transport protein involved in RCT were elevated in mice lack of apoE gene, which was substantially reduced by supplementing ellagic acid to Paigen diet-fed mice. In addition, ellagic acid attenuated hepatic lipid accumulation in apoE KO mice, evidenced by staining of hematoxylin and eosin and oil red O. Furthermore, the supplementation of 10 mg/kg ellagic acid favorably influenced the transcriptional levels of hepatic LDL receptor and scavenger receptor-B1 in Paigen diet-fed apoE KO mice. CONCLUSION: Ellagic acid may be an athero-protective dietary compound encumbering diet-induced atherogenesis though improving the RCT functionality.

Why A Multimedia Approach to English Education\ulcorner

  • Keem, Sung-uk
    • Proceedings of the KSPS conference
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    • 1997.07a
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    • pp.176-178
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    • 1997
  • To make a long story short I made up my mind to experiment with a multimedia approach to my classroom presentations two years ago because my ways of giving instructions bored the pants off me as well as my students. My favorite ways used to be sometimes referred to as classical or traditional ones, heavily dependent on the three elements: teacher's mouth, books, and chalk. Some call it the 'MBC method'. To top it off, I tried audio-visuals such as tape recorders, cassette players, VTR, pictures, and you name it, that could help improve my teaching method. And yet I have been unhappy about the results by a trial and error approach. I was determined to look for a better way that would ensure my satisfaction in the first place. What really turned me on was a multimedia CD ROM title, ELLIS (English Language Learning Instructional Systems) developed by Dr. Frank Otto. This is an integrated system of learning English based on advanced computer technology. Inspired by the utility and potential of such a multimedia system for regular classroom or lab instructions, I designed a simple but practical multimedia language learning laboratory in 1994 for the first time in Korea(perhaps for the first time in the world). It was high time that the conventional type of language laboratory(audio-passive) at Hahnnam be replaced because of wear and tear. Prior to this development, in 1991, I put a first CALL(Computer Assisted Language Learning) laboratory equipped with 35 personal computers(286), where students were encouraged to practise English typing, word processing and study English grammar, English vocabulary, and English composition. The first multimedia language learning laboratory was composed of 1) a multimedia personal computer(486DX2 then, now 586), 2) VGA multipliers that enable simultaneous viewing of the screen at control of the instructor, 3) an amplifIer, 4) loud speakers, 5)student monitors, 6) student tables to seat three students(a monitor for two students is more realistic, though), 7) student chairs, 8) an instructor table, and 9) cables. It was augmented later with an Internet hookup. The beauty of this type of multimedia language learning laboratory is the economy of furnishing and maintaining it. There is no need of darkening the facilities, which is a must when an LCD/beam projector is preferred in the laboratory. It is headset free, which proved to make students exasperated when worn more than- twenty minutes. In the previous semester I taught three different subjects: Freshman English Lab, English Phonetics, and Listening Comprehension Intermediate. I used CD ROM titles like ELLIS, Master Pronunciation, English Tripple Play Plus, English Arcade, Living Books, Q-Steps, English Discoveries, Compton's Encyclopedia. On the other hand, I managed to put all teaching materials into PowerPoint, where letters, photo, graphic, animation, audio, and video files are orderly stored in terms of slides. It takes time for me to prepare my teaching materials via PowerPoint, but it is a wonderful tool for the sake of presentations. And it is worth trying as long as I can entertain my students in such a way. Once everything is put into the computer, I feel relaxed and a bit excited watching my students enjoy my presentations. It appears to be great fun for students because they have never experienced this type of instruction. This is how I freed myself from having to manipulate a cassette tape player, VTR, and write on the board. The student monitors in front of them seem to help them concentrate on what they see, combined with what they hear. All I have to do is to simply click a mouse to give presentations and explanations, when necessary. I use a remote mouse, which prevents me from sitting at the instructor table. Instead, I can walk around in the room and enjoy freer interactions with students. Using this instrument, I can also have my students participate in the presentation. In particular, I invite my students to manipulate the computer using the remote mouse from the student's seat not from the instructor's seat. Every student appears to be fascinated with my multimedia approach to English teaching because of its unique nature as a new teaching tool as we face the 21st century. They all agree that the multimedia way is an interesting and fascinating way of learning to satisfy their needs. Above all, it helps lighten their drudgery in the classroom. They feel other subjects taught by other teachers should be treated in the same fashion. A multimedia approach to education is impossible without the advent of hi-tech computers, of which multi functions are integrated into a unified system, i.e., a personal computer. If you have computer-phobia, make quick friends with it; the sooner, the better. It can be a wonderful assistant to you. It is the Internet that I pay close attention to in conjunction with the multimedia approach to English education. Via e-mail system, I encourage my students to write to me in English. I encourage them to enjoy chatting with people all over the world. I also encourage them to visit the sites where they offer study courses in English conversation, vocabulary, idiomatic expressions, reading, and writing. I help them search any subject they want to via World Wide Web. Some day in the near future it will be the hub of learning for everybody. It will eventually free students from books, teachers, libraries, classrooms, and boredom. I will keep exploring better ways to give satisfying instructions to my students who deserve my entertainment.

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Molecular Epidemiologic Study of a Methicillin-resistant Staphylococcus aureus Outbreak at a Newborn Nursery and Neonatal Intensive Care Unit

  • Kang, Hyun Mi;Park, Ki Cheol;Lee, Kyung-Yil;Park, Joonhong;Park, Sun Hee;Lee, Dong-Gun;Kim, Jong-Hyun
    • Pediatric Infection and Vaccine
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    • v.26 no.3
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    • pp.148-160
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    • 2019
  • Purpose: This study aimed to investigate the molecular epidemiology of a methicillin-resistant Staphylococcus aureus (MRSA) outbreak at a newborn nursery and neonatal intensive care unit (NICU). Methods: During the outbreak, from August to September 2017, MRSA isolates collected from neonates and medical staff underwent genotyping and screened for virulence factors. Antibiotic susceptibilities were tested. Results: During the study period, 41 neonates were admitted at the nursery (n=27) and NICU (n=14). Of these, 7 had MRSA infections (skin infection [n=6] and sepsis [n=1]) and 4 were colonized with MRSA. Associated medical staff (n=32) were screened; three were nasal MRSA carriers. Staphylococcal chromosomal cassette mec (SCCmec) type II, sequence type (ST) 89, spa type t375 was found to be the skin infection outbreak causing strain, with multi-drug resistance including low-level mupirocin resistance. SCCmec type IVa, ST 72, and a novel spa type designated t17879, was the cause of MRSA sepsis. Many different types of MRSA were colonized on the neonates; however, SCCmec type IVa, ST 72, spa type t664 was colonized in both neonates and a NICU nurse. All MRSA isolates from colonized infants were positive for the Panton-Valentine leukocidin (PVL) toxin gene. Conclusions: The strain causing an outbreak of skin infections had multi-drug resistance. Also, MRSA colonized in the neonates were found to carry the PVL toxin gene. Because different strains are present during an outbreak, molecular epidemiologic studies are important to identify the outbreak strain and colonized strains which aid in effective control and prevention of future MRSA outbreaks.

Use of Human Serum Albumin Fusion Tags for Recombinant Protein Secretory Expression in the Methylotrophic Yeast Hansenula polymorpha (메탄올 자화효모 Hansenula polymorpha에서의 재조합 단백질 분비발현을 위한 인체 혈청 알부민 융합단편의 활용)

  • Song, Ji-Hye;Hwang, Dong Hyeon;Oh, Doo-Byoung;Rhee, Sang Ki;Kwon, Ohsuk
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.17-25
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    • 2013
  • The thermotolerant methylotrophic yeast Hansenula polymorpha is an attractive model organism for various fundamental studies, such as the genetic control of enzymes involved in methanol metabolism, peroxisome biogenesis, nitrate assimilation, and resistance to heavy metals and oxidative stresses. In addition, H. polymorpha has been highlighted as a promising recombinant protein expression host, especially due to the availability of strong and tightly regulatable promoters. In this study, we investigated the possibility of employing human serum albumin (HSA) as the fusion tag for the secretory expression of heterologous proteins in H. polymorpha. A set of four expression cassettes, which contained the methanol oxidase (MOX) promoter, translational HSA fusion tag, and the terminator of MOX, were constructed. The expression cassettes were also designed to contain sequences for accessory elements including His8-tag, $2{\times}(Gly_4Ser_1)$ linkers, tobacco etch virus protease recognition sites (Tev), multi-cloning sites, and strep-tags. To determine the effects of the size of the HSA fusion tag on the secretory expression of the target protein, each cassette contained the HSA gene fragment truncated at a specific position based on its domain structure. By using the Green fluorescence protein gene as the reporter, the properties of each expression cassette were compared in various conditions. Our results suggest that the translational HSA fusion tag is an efficient tool for the secretory expression of recombinant proteins in H. polymorpha.

Evaluation of Usefulness of Portal Image Using Electronic Portal Imaging Device (EPID) in the Patients Who Received Pelvic Radiation Therapy (골반강 내 방사선 치료 환자에서 Electronic Portal Imaging Device(EPID)를 이용한 Portal Image의 유용성에 관한 연구)

  • Kim Woo Chul;Park Won;Kim Heon Jong;Park Seong Young;Cho Young Kap;Loh John J;Suh Chang Ok;Kim Gwi Eon
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.497-504
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    • 1998
  • Purpose : To evaluate the usefulness of electronic portal imaging device through objective compare of the images acquired using an EPID and a conventional port film Materials and Methods : From Apr. to Oct. 1997, a total of 150 sets of images from 20 patients who received radiation therapy in the pelvis area were evaluated in the Inha University Hospital and Severance Hospital. A dual image recording technique was devised to obtain both electronic portal images and port film images simultaneously with one treatment course. We did not perform double exposure five to ten images were acquired from each patient. All images were acquired from posteroanterior (PA) view except images from two patients. A dose rate of 100-300 Mu/min and a 10-MV X-ray beam were used and 2-10 MUs were required to produce a verification image during treatment. Kodak diagnostic film with metal/film imaging cassette which was located on the top of the EPID detector was used for the port film. The source to detector distance was 140 cm. Eight anatomical landmarks (pelvic brim, sacrum, acetabulum. iliopectineal line, symphysis, ischium, obturator foramen, sacroiliac joint) were assessed. Four radiation oncologist joined to evaluate each image. The individual landmarks in the port film or in the EPID were rated - very clear (1), clear (2), visible (3), not clear (4), not visible (5). Results : Using an video camera based EPID system. there was no difference of image quality between no enhanced EPID images and port film images. However, when we provided some change with window level for the portal image, the visibility of the sacrum and obturator foramen was improved in the portal images than in the port film images. All anatomical landmarks were more visible in the portal images than in the port film when we applied the CLAHE mode enhancement. The images acquired using an matrix ion chamber type EPID were also improved image qualify after window level adjustment. Conclusion : The quality of image acquired using an electronic portal imaging device was comparable to that of the port film. When we used the enhance mode or window level adjustment. the image quality of the EPID was superior to that of the port film. EPID may replace the port film.

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