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The antimicrobial resistance rate in bacteria has increased over the last several decades. The transfer of antimicrobial resistant determinants on mobile genetic elements could cause the accelerated emergence and spread of multidrug resistant bacteria. This study investigated the aminoglycoside resistance determinants transferred by mobile genetic elements in a total of 33 aminoglycoside non-susceptible E. coli isolated from clinical specimens in Chungcheong province. 16S ribosomal RNA methyl-transferases (RMTases) and aminoglycoside-modifying enzyme (AME) genes were detected via PCR and DNA sequencing. The most common AME genes were aac(3′)-II gene (54.5%), followed by aph(3′)-Ia (18.2%) and aac(6′)-Ib (15.2%). None of the evaluated RMTase genes were detected in the 33 isolates. Seventeen of the 18 isolates harboring aac(3′)-II gene were resistant to gentamicin, and 16 of them were resistant to tobramycin. The 5 isolates harboring aac(6′)-Ib gene were all resistant to tobramycin. In this study, we confirmed that one of the important mechanisms of aminoglycoside resistance in E. coli isolated from human is the acquisition of AME genes. Continuing investigations of antimicrobial resistant determinants in bacteria isolated from human may be required to prevent dissemination of antimicrobial resistant bacteria.

Objectives: This study aimed to test the effect of photodynamic therapy (PDT) on the inhibition and removal of biofilms containing multidrug-resistant Acinetobacter baumannii.Methods: Using multidrug-resistant A. baumannii strains, an antibiotic susceptibility test was performed using the Gram-negative identification card of the Vitek 2 system (bioMérieux Inc., France), as well as an analysis of resistance genes, the effects of treatment with a light-emitting diode (LED) array using Radachlorin (RADA-PHARMA Co., Ltd., Russia), and transmission and scanning electron microscopy to confirm the biofilm-inhibitory effect of PDT.Results: The antibiotic susceptibility test revealed multiple resistance to the antibiotics imipenem and meropenem in the carbapenem class. A class-D–type β-lactamase was found, and OXA-23 and OXA-51 were found in 100% of 15 A. baumannii strains. After PDT using Radachlorin, morphological observations revealed an abnormal structure due to the loss of the cell membrane and extensive morphological changes, including low intracellular visibility and small vacuoles attached to the cell membrane.Conclusion: PDT involving a combination of LED and Radachlorin significantly eliminated the biofilm of multidrug-resistant A. baumannii. Observations made using electron microscopy showed that PDT combining LED and Radachlorin was effective. Additional studies on the effective elimination of biofilms containing multidrug-resistant bacteria are necessary, and we hope that a treatment method superior to sterilization with antibiotics will be developed in the future.

Acinetobacter baumannii는 광범위한 항생제에 대한 저항성으로 인해 감염된 환자의 사망률이 높아지는 적색 경보 병원체로 분류됩니다. 이 연구에서 다제 내성 A. baumannii(MRAB)의 18가지 임상 분리 균주에 대해 일부 에센셜 오일(티트리, 로즈마리, 라벤더 오일)의 항균 활성을 평가하고자 하였다. Carbapenemase 선별을 위한 Hodge 시험법은 A. baumannii의 20 가지 균주가 모두 imipenem에 내성이 있음을 보여주었습니다. 다제 내성 미생물의 확인은 VITEK 시스템을 통해 수행 하였다. 에센셜 오일의 항균 활성은 MRAB에 대한 디스크 확산 방법으로 평가하였다. 디스크 확산 방법에서 tee tree는 라벤더 오일에 비해 억제 크기가 가장 크게 증가했으며, 로즈마리는 항균 효과가 없었다. 티 트리 오일은 가장 일반적인 인간 병원균 및 MRAB 감염의 치료 및 예방을 위한 대체 천연 제품으로 유용할 것으로 보인다. 따라서 이 연구의 결과는 다제 내성 A. baumannii의 항균 효과를 입증했으며, 미래에 천연 에센셜 오일을 사용하는 손 소독제와 같은 항균제로 사용될 것으로 예상 됩니다.

임상병리학과 학제 단일화에 대한 의견을 교수와 병원근무자들을 대상으로 설문조사 형식으로 조사하여 분석하였다. 2018년 1월 18일부터 25일까지 웹 버전의 설문지가 255명의 대학교수들과 4,000명의 병원근무자들에게 배포되었다. 회수율은 교수들은 79명(30.9%), 병원실무자들은 1,368명(34.2%)이 응답하였다. 임상병리학의 학제 이원화에 대한 인식조사에서는 교수와 병원근무자 모두 4년제로 학제 통합이 필요하다고 조사되었다. 4년제 통합 전제조건은 임상병리 인증평가원 설립과 교육과정의 표준화가 높게 조사되었고, 4년제 통합의 필요조건은 학생정원조정과 교육과정의 표준화가 높게 조사되었다. 4년제 통합방식은 인증평가를 통과한 대학만이 4년제를 운영해야 한다는 의견이 교수와 병원근무자 모두 가장 높게 조사되었다. 4년제 통합시 대학정원 적정인원은 40명 미만이 가장 높게 조사되었다. 결론적으로 교수들과 병원실무자들은 의료환경의 급격한 변화에 부응하는 전문성을 가진 임상병리사를 배출하기 위해서는 임상병리학과 학제의 4년제 통합이 필수적이고, 이를 위해 임상병리인증평가원 설립, 교육과정의 표준화, 학생정원조정 이 전제되어야 한다고 생각하고 있음을 보여준다.

식물로부터 유래되는 휘발성 유기 화합물인 에센셜 오일은 인체에 약리효과를 나타낸다고 알려져 있다. 본 연구에서는 화농성 질환과 병독성 질환 비뇨기계통 감염에 나올 수 있는 병원성 세균 Staphylococcus aureus (그람양성알균), Escherichia coli (그람음성막대균), Pseudomonas aeruginosa (그람음성막대균, 극성단모균)을 대상으로 천연 허브 오일 중에 비교적 많이 알려져 있는 라벤더(lavender), 티트리(tea tree), 로즈마리(rosemary), 멜리사(melisa) 4종류의 단일 오일과 비교해, 티트리와 로즈마리, 멜리사를 블렌딩하였을 때 항균력이 상승되는 효과가 있는지 연구하였다. 그 결과 디스크 확산법에서 블렌딩오일(티트리, 로즈마리, 멜리사)의 경우 단독의 허브오일보다 S. aureus에서18 mm, E. coli 에서 22 mm 와 P. aeruginosa는13mm의 우수한 항균능력을 확인할 수 있었다. 뿐만 아니라 블렌딩 오일의 경우 최소억제농도(Minimum Inhibitory Concentrations, MIC) 분석에서 S. aureus, E. coli 에서는 0.3125%에서, P. aeruginosa는 1.25%에서 성장억제효과가 확인되었다. 따라서 본 실험에서는 블렌딩 오일이 Oxacillin과 로즈마리, 티트리보다 항균효과가 우수한 것으로 증명되어 단독의 허브오일보다 블렌딩 오일이 우수한 항균력을 나타나는 것을 확인할 수 있었다. 그러므로 허브의 블렌딩을 활용하면 다제 내성균에 대한 항균시너지효과를 발휘할 수 있음을 입증했을 뿐만 아니라 향후 천연보존제와 건강기능성 식품 및 화장품에 활용 가능성을 기대해본다.

This study analyzed the distribution of the blood glucose level according to the fasting status. Moreover, a relationship was analyzed between fasting blood glucose level and glycemic control indicators. A total of 707 outpatients, who visited Dankook University Hospital, were included and classified into either the fasting group and the non-fasting group. The mean blood glucose level of each group was calculated and analyzed by sex, age, and clinic. In addition, blood glucose, HbA1c, fructosamine, and 1,5-AG were measured in 153 fasting health check-up patients, and the correlation between the blood glucose level and glycemic control indicators was evaluated. Blood glucose averages between the two groups (non-fasting 111.9 vs. fasting 103.6 mg/dL) were different (p＜0.05); and the mean difference was lower in women (4.8 mg/dL) than in men (12.2 mg/dL). A significant difference of the median glucose values among the age groups was only observed in the non-fasting group (Kruskal-Wallis test, p＜0.01), and not in the fasting group. A 1,5-Anhydroglucitol was estimated to be significantly correlated with the fast blood glucose level in the range of the criteria of impaired fasting glucose (IFG). We presented an assessment of the distribution of blood glucose level in accordance with the fasting status among outpatients, and estimated that 1,5-anhydroglucitol was well correlated with the fasting blood glucose than fructosamine and HbA1c, through the analysis of results of health screening subjects. It is suggested that the use of glycemic indicators that reflect short-term blood glucose control can be used together with the blood glucose measurement in the screening of diabetes mellitus.

Secondary metabolism in actinomycetes has been known to be controlled by a small molecule, -butyrolactone autoregulator, the binding of which to each corresponding receptor leads to the regulation of the transcriptional expression of the secondary metabolites. We expected that expression of an autoregulator receptor or a pleiotropic regulator in a non-host was to be gained insight of effective production of new metabolic materials. In order to study the function of the receptor protein (seaR), which is isolated from Saccharopolyspora erythraea, we introduced the seaR gene to Streptomyces coelicolor A3(2) as host strains. An effective transformation procedure for S. coelicolor A3(2) was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a -derived integration vector, pSET152, which contained int, oriT, attP and (erythromycin promotor). Therefore, the pEV615 was introduced into S. coelicolor A3(2) by conjugation and integrated at the attB locus in the chromosome of the recipients by the integrase (int) function. Exconjugant of S. coelicolor A3(2) containing the seaR gene was confirmed by PCR and transcriptional expression of the seaR gene in the transformant was analyzed by RT-PCR. In case of S. coelicolor A3(2), a phenotype microarray was used to analyze the phenotype of transformant compared with wild type by seaR expression. After that, in order to confirm the accuracy of the results obtained from the phenotype microarray, an antimicrobial susceptibility test was carried out. This test indicated that sensitivity of the transformant was higher than wild type in tetracycline case. These results indicated that some biosynthesis genes or resistance genes for tetracycline biosynthesis in transformant might be repressed by seaR expression. Therefore, subsequent experiments, analysis of transcriptional pattern of genes for tetracycline production or resistance, are needed to confirm whether biosynthesis genes or resistance genes for tetracycline are repressed or not.

Photodynamic therapy (PDT) apply photosensitizers and light. The purpose of this study was to evaluate the in vitro efficacy of PDT using blue LED (light emitting diode) with photofrin and radachlorin for Propionibacterium acnes. The colony forming units method was used to assess the antibacterial activity. Suspension (1 mL) containing P. acnes at 1×105 CFU/mL were prepared and then 2 fold serial diluted to 12.5 g/mL from 50 g/mL concentration of photofrin and radachlorin. After 60 minutes incubation, light was irradiated for 10 to 30 minutes using the following light source of wavelength 460 nm, each energy density 36, 72 and 108 J/cm2. Bacterial growth was evaluated after 72 hours incubation in a Phenylethanol Blood Agar (PEBA) culture. In addition, flow cytometric analysis were performed to measure the live cell after PDT. Also transmission electron microscopy (TEM) was employed to evaluate the effect of pathogens by PDT. The PDT Group was perfectly killed to all kind of photosensitizers dose of 12.5 g/mL with irradiation of 10 minutes. Also other Groups were killed to all kind of photosensitizers dose of 6.25 g/mL with irradiation time of 20 and 30 minutes. The flow cytometry showed a lower number of viable bacteria in the PDT group compared to the control group. The images of the TEM results were showed in cytoplasmic membrane damage and partially deformed to cell morphologies. These results suggest that radachlorin and photofrin combine blue LED PDT can be effectively treated when was proved treatment for acnes therapy.

Lateral canal benign paroxysmal vertigo (BPPV) causing dizziness is a common cause is not found while continuing to appeal for vertigo is a typical disease. It is characterized by acute stand up, brief and rotatory vertigo attacks provoked by change in head position. Treatment requires only one treatment visit in most patients. However, there are significant numbers of patients who require multiple treatment visits for relief. The purpose of this study benign paroxysmal positional vertigo treatment of type affect is to analyze the cause. Dizziness and vertigo patient`s in patients admitted to the dizziness center of lateral canal benign paroxysmal positional vertigo were classified. In patients with lateral canal benign paroxysmal positional vertigo and accompanying lateral 15 treatment affects disease were investigated. March 2008 to November 2010 lateral canal benign paroxysmal positional vertigo 166 people cure rate of patients was investigated. First time the success rate of 74.1%, twice times the success rate of 12.0%, three times the success rate of 9.6%, more than three times the success rate was 4.2%. Affecting factor treatment of benign paroxysmal positional vertigo in post-traumatic, medicine disease, headache, cerebral infarction, small vessel disease, vestibulopathy, (p＜0.05). Statistical analysis using SPSS (version 12K) in coefficient measure through descriptive statistical of cross table.

Twenty-four male Sprague-Dawley rats were randomly divided into 1 ㎃(0.11 ㎃/㎠), 2 ㎃(0.22 ㎃/㎠), 4 ㎃(0.44 ㎃/㎠), and 8 ㎃(0.89 ㎃/㎠) groups. These rats were exposed to anodic iontophoresis of 1% MB using a direct current for 15 minutes. The penetration depth were measured using light microscopy from cryosections of skin tissue. The tissue concentration and transdermal transport were measured using biochemical analysis from target skin tissues. The data were analyzed with one-way analysis of variance.

목적: 본 연구는 저강도 맥동 초음파적용이 흰쥐의 전층 상처 치유와 아교질 축적 및 아교질 mRNA발현에 미치 는 영향을 규명할 목적으로 시행하였다. 방법: 12마리의 Sprague-Dawley계 흰쥐를 저강도 맥동 초음파군 (n=6)과 대조군 (n=6)에 무작위 배정하고 등에 19.63 mm2 크기의 전층 적출 상처를 만든 다음 저강도 맥동 초음파군은 3 MHz, 순환주기 20%, SATA 강도 0.4 W/cm2 로 1일 1회, 1회 5분씩 초음파를 적용하고, 대조군은 가짜 초음파를 적용하였다. 7일간 처치 후 초음파군과 대조군 의 아교질 축적, 아교질 mRNA 발현, 상처치유율, 절반치 유시간을 비교하였다. 결과 : 초음파군의 아교질 축적 (p

The fluorescence detection of intracellular metal ions are high interest in the fields of organic molecular chemistry and cellular biology. This study was purposed to detection for mercury and zinc in the cell using fluorescent chemosensor (FS). FS exhibits a weak fluorescence, but emits strong fluorescence upon Zn

Objectives/Hypothesis: The aim of this study was to evaluate the antibacterial effects of photodynamic therapy (PDT) on common bacteria causing otitis media with effusion (OME). Methods: An in vitro study was carried out using a hematoporphyrin derivative sensitizer (Photogem; Lemonosov Institute of Fine Chemical, Moscow, Russia) and a 632-nm diode laser on Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae. The presence of colony-forming units of the bacteria was examined, the microscopic structures of the bacteria were examined by transmission electron microscopy (TEM), and flow cytometry of the bacteria was performed. An in vivo PDT study was performed using gerbils. S. pneumoniae or H. influenzae were injected into bullae. The Photogem was injected into the bullae 2 days later when OME developed, and transcanal irradiation with the 632-nm diode laser (90 J) was performed. Middle ear and bulla were washed with Dulbecco's phosphate buffered saline (DPBS) and the washed DPBS was cultured. The presence of bacterial colonies was examined. Results: The PDT was effective in killing all three kinds of bacteria. TEM showed damaged bacterial cell membranes and cytoplasmic structures, and the flow cytometry showed a lower number of viable bacteria in the PDT group compared to the control group. PDT was effective in killing S. pneumoniae in 87% of the infected bullae with OME, whereas it was effective in eradicating H. influenzae in 50% of the infected bullae with OME. Conclusions: The results of these studies demonstrated that PDT may be effective to treat otitis media. PDT may have clinical implications in the treatment of otitis media that is resistant to antibiotic therapy. Laryngoscope, 2009

The fluorescence of NABQ increases remarkably in the presence of Zn(2+) and is quenched by Hg(2+). As shown by confocal imaging, NABQ-Zn(2+) can penetrate cells, where the bound Zn(2+) is exchanged for Hg(2+). This results in the concomitant export of Hg(2+) from the cells, showing that NABQ can act as a Zn(2+) carrier and as a Hg(2+) extracting agent in living cells.

We present the design, synthesis, spectroscopic properties, and biological evaluation of a single galactose-appended naphthalimide (1). Probe 1 is a multifunctional molecule that incorporates a thiol-specific cleavable disulfide bond, a masked phthalamide fluorophore, and a single galactose moiety as a hepatocyte-targeting unit. It constitutes a new type of targetable ligand for hepatic thiol imaging in living cells and animals. Confocal microscopic imaging experiments reveal that 1, but not the galactose-free control system 2, is preferentially taken up by HepG2 cells through galactose-targeted, ASGP-R-mediated endocytosis. Probe 1 displays a fluorescence emission feature at 540 nm that is induced by exposure to free endogenous thiols, most notably GSH. The liver-specificity of 1 was confirmed in vivo via use of a rat model. The potential utility of this probe in indicating pathogenic states and as a possible screening tool for agents that can manipulate oxidative stress was demonstrated in experiments wherein palmitate was used to induce lipotoxicity in HepG2 cells.

The aim of this study was to evaluate the antibacterial effects of photodynamic therapy (PDT) on common bacteria causing otitis media with effusion (OME).An in vitro study was carried out using a hematoporphyrin derivative sensitizer (Photogem; Lemonosov Institute of Fine Chemical, Moscow, Russia) and a 632-nm diode laser on Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae. The presence of colony-forming units of the bacteria was examined, the microscopic structures of the bacteria were examined by transmission electron microscopy (TEM), and flow cytometry of the bacteria was performed. An in vivo PDT study was performed using gerbils. S. pneumoniae or H. influenzae were injected into bullae. The Photogem was injected into the bullae 2 days later when OME developed, and transcanal irradiation with the 632-nm diode laser (90 J) was performed. Middle ear and bulla were washed with Dulbecco's phosphate buffered saline (DPBS) and the washed DPBS was cultured. The presence of bacterial colonies was examined.The PDT was effective in killing all three kinds of bacteria. TEM showed damaged bacterial cell membranes and cytoplasmic structures, and the flow cytometry showed a lower number of viable bacteria in the PDT group compared to the control group. PDT was effective in killing S. pneumoniae in 87% of the infected bullae with OME, whereas it was effective in eradicating H. influenzae in 50% of the infected bullae with OME.The results of these studies demonstrated that PDT may be effective to treat otitis media. PDT may have clinical implications in the treatment of otitis media that is resistant to antibiotic therapy.

A novel coumarin-based fluorogenic probe bearing the 2-picolyl unit (1) was developed as a fluorescent chemosensor with high selectivity and suitable affinity in biological systems toward Cu(2+) over other cations tested. The fluorescence on-off mechanism was studied by femtosecond time-resolved fluorescence (TRF) upconversion technique and ab initio calculations. The receptor can be applied to the monitoring of Cu(2+) ion in aqueous solution with a pH span 4-10. To confirm the suitability of 1 for biological applications, we also employed it for the fluorescence detection of the changes of intracellular Cu(2+) in cultured cells. The results indicate that 1 should be useful for the fluorescence microscopic imaging and the study on the biological functions of Cu(2+).

Background Tuberculosis (TB) remains an important cause of morbidity and mortality throughout the world. The surge of TB has been accompanied by an increase in multi-drug-resistant tuberculosis (MDR-TB). In this study, we developed a denaturing HPLC (DHPLC) method for detecting rpoB gene mutation as a rifampin resistance based on sequence. Methods In this study, we used 99 mycobacterial isolates grown in Ogawa media. At first, we used a PCR method that can amplify the 235 bp and 136 bp rpoB DNAs of Mycobacterium tuberculosis complex (MTB) and Non-tuberculous mycobacteria (NTM). And then, PCR-restriction fragment length polymorphism (RFLP) of rpoB DNA (342 bp), which comprises the Rif(T) region, was used for the differential identification of Mycobacteria. Finally, we detected these amplicons by DHPLC, compared to PCR-RFLP results, and performed sequencing. Results Among 99 mycobacterial isolates, 80 (81%) were MTB and 19 (19%) were NTM. NTM were identified to 7 different species by DHPLC and PCR-RFLP. rpoB mutation was detected in 9 (11%) of the MTB specimens. These results were confirmed by using sequencing. Conclusions DHPLC provided a rapid, simple, and automatable performance for detection of rifampin resistant Mycobacterium tuberculosis complex and would be helpful as a supplemental method in high-throughput clinical laboratories.

The aim of this study was to evaluate antibacterial effects of PDT on common bacteria causing otitis me dia with effusion (OME). In vitro study was carried out using a hematoporphyrin derivative sensitizer (photogem) and 632 nm diode laser on H. influenzae, M. catarrhalis, and S. pneumoniae, . One ml of each bacterial suspension was incubated for 3 hours and various concentrations of photogem were administered into the suspension. The suspensions were irradiated with 632 diode laser (15 J/cm 2 ). The presence of colony forming units of the bacteria was examined, microscopic structures of bacteria were examined by TEM, and cytometry of bacter ia was performed. The PDT was effective in killing all 3 kinds of bacteria. TEM showed damaged bacterial cell membrane and cytoplasmic structures and the flow cytometr y showed lower number of viable bacteria in PDT group comparing to the control group. In vivo PDT study was performed using gerbil. S. pneumoniae or H. influenzae was injected into bullae. Photogem was injected into bullae in 2 days by when OME was developed and transcanal irradiation of 632 nm diode laser (90 J) was performed with a fiber perforated through an ear drum into a middle ear cavity and bulla. Four days after PDT, middle ear and bulla were washed with DPBS and the washed DPBS was cultured. The presence of bacterial col onies was examined. PDT was effective in killing S. pneumoniae in 87 % of the infected bullae with OME while it was effective to eradicate H. influenzae in 50 % of the infected bullae with OME. The results of these studies demonstrated that PDT may be effec tive to treat otitis media. It may have clinical implication to treat otitis media that is resistant to antibiotic therapy. Key Words: Otitis Media, Photodynamic Therapy, Photosensitizer, Diode Laser, OME, S. pneumonia, H. in fluenza, M. catarrhalis

A rationally designed, coumarin-based fluorescent sensor imino-coumarin (IC) displays high selectivity for Cu(2+) over a variety of competing metal ions in aqueous solution with a significant fluorescence increase. DFT/TDDFT calculations support that the fluorescence 'turn-on' of IC originates from blocking the electron transfer of the nitrogen lone pair upon complexation with Cu(2+). IC was successfully applied to microscopic imaging for detection of Cu(2+) in LLC-MK2 cells (in vitro) and several living organs (in vivo).

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most prevalent dermatology pathogens in hospitals and increasingly recognized in communities. We determined PFGE pattern of SmaI-restricted genomic DNA, coagulase type, and antimicrobial susceptibility of MRSA isolated in 2008 from dermatology inpatients and healthy hospital employees in A Hospital and from primary school children in Iksan city, Korea. Overall, the isolation rate of MRSA was 3.8% from the 788 normal persons: 4.9% from hospital employees and 1.1% from primary school children. MRSA was isolated in six of 13 (46.2%) family members of four school children with MRSA. The most prevalent coagulase serotype was Ⅱ from patients and V from healthy individuals. Ten of twenty and six of twenty MRSA isolates from patients and from healthy personnel, respectively, had identical PFGE patterns, suggesting that these are originated from identical clones. Against MRSA from patients, only vancomycin was the most active (MIC range ≤2 μg/㎖), whereas the resistance rates were 35% to rifampin and 65% to mupirocin. The resistance rates of patient isolates were ≥90% to amikacin, clindamycin, ciprofloxacin, erythromycin, fusidic acid, gentamicin and tetracycline. In conclusion, the MRSA carriage rates of healthy hospital workers were relatively high, 2.3~7.7%, depending on groups. Family members of a few primary school children with MRSA showed a high carriage rate, suggesting that intrafamily transmission occurred. MRSAs isolated from dermatology inpatients were relatively more resistant to various antimicrobial agents, including mupirocin, but all isolates were susceptibility to vancomycin.