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Improving the Rapidity and Accuracy of Carbapenem-Resistant Enterobacteriaceae Detection by Shortening the Enrichment Duration

  • Miyoung, Lee
    • Microbiology and Biotechnology Letters
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    • v.50 no.4
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    • pp.584-591
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    • 2022
  • Identifying carbapenem-resistant Enterobacteriaceae (CRE) is necessary to prevent nosocomial CRE infection outbreaks. Here, a rapid identification method with reduced enrichment time was developed without compromising accuracy. A total of 49 rectal swabs requested for CRE screening at the Department of Diagnostic Medicine at Hospital B in Busan, Korea, were included in this study. Specimens were inoculated on MacConkey and CHROMID Carba media either directly or following enrichment for 3, 6, and 24 h in 100 μl trypticase soy broth containing an ertapenem disk. The enriched cultures were further inoculated on CHROMID Carba or MacConkey media containing an ertapenem disk. In total, 19 CRE and 5 carbapenem-intermediate Enterobacteriaceae isolates were obtained from the 49 swabs. Among the 19 CRE isolates, carbapenemase-producing Enterobacteriaceae constituted 13 strains. Moreover, of the 19 CRE isolates, 16 (81.25%) and 17 (88.24%) were identified from the direct cultures on MacConkey and CHROMID Carba media, respectively. After 3 h of enrichment, the proportions of the CRE identified in the media were: MacConkey medium, 16/19 (81.25%); CHROMID Carba medium, 17/19 (88.24%); and MacConkey medium containing an ertapenem disk, 17/19 (88.24%). The detection rates after 6 h of enrichment were the same for all three media (19/19 strains, 100%), whereas those after 24 h of enrichment were 21, 22, and 24 strains, respectively, but included false positives. These findings suggest that a 6-h enrichment before inoculation on the CHROMID Carba medium is optimal for the rapid and accurate detection of CRE in clinical samples.

Risk Factors of Carbapenem-resistant Enterobacteriaceae Acquisition at a Community-based Hospital (지역기반 상급종합병원 내 카바페넴 내성 장내세균 획득에 관한 위험인자)

  • Lee, Yeonju;Kang, Ji Eun;Ham, Jung Yeon;Lee, Ja Gyun;Rhie, Sandy Jeong
    • Korean Journal of Clinical Pharmacy
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    • v.30 no.2
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    • pp.120-126
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    • 2020
  • Objective: The rising number of carbapenemase-resistant Enterobacteriaceae (CRE) cases has become a concern worldwidely. This study investigated patient characteristics with CRE and analyzed the risk factors associated with its acquisition. Methods: A retrospective review of the electronic medical records of the Kangbuk Samsung Medical Center from May 2016 to April 2019 was performed. The inclusion criterion was hospitalized patients aged ≥18 years with confirmed CRE acquisition. Patients were divided by CRE acquired and non-required patients. CRE acquired patients were those with CRE confirmed by their active surveillance cultures, while non-acquired patients were those with carbapenemase-sensitive Enterobacteriaceae (CSE). If CRE was isolated more than once during hospitalization, only the first isolation was used for data analysis. Patient characteristics, antibiotic used, and the duration of use were compared between two groups using univariate analysis, and the risk factors associated with CRE were analyzed using multiple logistic regression analysis. Results: Among the 73 CRE acquired patients, 44 (60.3%) were positive for carbapenemase-producing Enterobacteriaceae (CPE). Infection from Klebsiella pneumonia (42 cases, 57.5%), Escherichia coli (17 cases, 23.3%), and Enterobacter cloacae (5 cases, 6.8%). The risk of CRE acquisition was significantly increased by 4.99 times [confidence interval (CI), 1.40-17.78; p=0.013] with mechanical ventilation, 3.86 times (CI, 1.59-9.36; p=0.003) with penicillin administration, and 21.19 times (CI, 6.53-68.70; p<0.001) with carbapenem administration. Conclusions: Proper antibiotic use including the selection, frequency, and duration, and patients on mechanical ventilators need close monitoring.

Anti-parkinsonian effect of Cyperi Rhizoma via inhibition of neuroinflammatory action (향부자(香附子)의 염증 억제 작용을 통한 항파킨슨 효과)

  • Kim, Hyo Geun;Sim, Yeomoon;Oh, Myung Sook
    • The Korea Journal of Herbology
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    • v.28 no.5
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    • pp.21-28
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    • 2013
  • Objectives : The aim of this study was to investigate the neuroprotective effects and mechanisms of Cyperi Rhizoma extracts (CRE) using in vitro and in vivo models of Parkinson's disease (PD). Methods : We evaluated the neuroprotective effect of CRE against 1-methyl-4-phenylpyridinium (MPP+) toxicity using tyrosine hydroxylase immunohistochemistry (IHC) in primary rat mesencephalic dopaminergic neurons. In addition, the effect of CRE was evaluated in mice PD model induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). For evaluations, C57bl/6 mice were orally treated with CRE 50 mg/kg for 5 days and were injected intraperitoneally with MPTP (20 mg/kg) at 2 h intervals on the last day. To identify the CRE affects on MPTP-induced neuronal loss of dopaminergic neurons in substantia nigra pars compacta (SNpc) and striatum of mice, the behavioral tests and IHC analysis were carried out. Also, we conducted nitric oxide (NO) and tumor necrosis factor-alpha (TNF-${\alpha}$) assay in dopaminergic neurons and IHC using glial markers in SNpc of mice to assess the anti-inflammation effects. Results : In primary mesencephalic culture system, CRE protected dopaminergic cells against $10{\mu}M$ MPP+-induced toxicity at 0.2 and $1.0{\mu}g/mL$. In the behavior tests, CRE treated group showed improved motor deteriorations than those in the MPTP only treated group. CRE significantly protected striatal dopaminergic damage from MPTP-induced neurotoxicity in mice. Moreover, CRE inhibited productions of NO and TNF-${\alpha}$ in dopaminergic culture system and activation of astrocyte and microglia in SNpc of the mice. Conclusion : We concluded that CRE shows anti-parkinsonian effect by protecting dopaminergic neurons against MPP+/MPTP toxicities through anti-inflammatory actions.

Risk factors for the colonization of carbapenem-resistant Enterobacteriaceae in patients transferred to a small/medium-size hospital in Korea: a retrospective study (중소병원으로 전원 온 환자의 카바페넴내성장내세균속균종 보균 위험요인)

  • Misun Lee;Hyunjung Kim
    • Journal of Korean Biological Nursing Science
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    • v.25 no.4
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    • pp.285-294
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    • 2023
  • Purpose: This study aimed to identify the colonization rate of carbapenem-resistant Enterobacteriaceae (CRE), the characteristics of CRE isolates, and risk factors for CRE colonization in patients transferred to the general wards of a small/medium-sized hospital. Methods: This retrospective study was conducted on patients who underwent CRE culture tests within 24 hours of admission among patients transferred to a small/medium-sized hospital. Forty-seven patients confirmed as positive for CRE were classified as belonging to the patient group. For the control group, 235 patients (five times the number of the patient group) were matched by sex, age, and diagnosis, and then selected at random. Data were analyzed using descriptive analysis and multiple logistic regression analysis. Results: The CRE colonization rate was 5% (47 out of 933 patients), and Klebsiella pneumoniae (68.0%) was the most common isolate of CRE. The positivity rate of carbapenemase-producing Enterobacteriaceae was 61.7%. The risk factors for CRE colonization included renal disease (odds ratio [OR]=4.93; 95% confidence interval [CI], 1.49-16.31), heart disease (OR=3.86; 95% CI, 1.35-11.01), indwelling urinary catheters (OR=4.43; 95% CI, 1.59-12.36), and cephalosporin antibiotic use (OR=8.57; 95% CI, 1.23-59.60). Conclusion: Having a comorbid renal or cardiac disease, an indwelling urinary catheter, or a history of exposure to cephalosporin antibiotics could be classified as risk factors for CRE colonization in patients transferred to small and medium-size hospitals. It is necessary to perform active infection control through proactive CRE culture testing of patients with risk factors.

Catabolite Repression of the Bacillus stearothermophilus $\beta$-Xylosidase Gene (xylA) in Bacillus subtilis

  • Cho, Ssang-Goo;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.8 no.1
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    • pp.21-27
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    • 1998
  • The xylA gene of Bacillus stearothermophilus encoding the major ${\beta}$-xylosidase was previously cloned and sequenced. In the present study we examined the regulation of the cloned xylA gene expression in Bauillus subtilis MW15 carrying the xylA::aprA fusion plasmids. The induction of the fused xylA gene expression remained uninfluenced by any of the carbon sources tested but the gene expression was repressed about 2-3 fold in the presence of glucose. Two CRE-like sequences (CRE-1: nucleotides + 124 to +136 and CRE-2: +247 to +259) were recognized within the reading frame region of the xylA gene. The deletion experiments showed that the CRE-2 sequence had a role in catabolite repression (CR) as a true CRE of the xylA gene, but the CRE-1 had no effect on CR of the xylA gene expression. Surprisingly, the deletion of the CRE- 1 sequence reduced about 2~3 fold of the expression of the xylA fused gene. The repression ratios of the xylA gene expression were estimated to be about 0.4 from the assay of subtilisin activity, and about 0.3 at the level of transcription by determining the amounts of xylA transcripts in B. subtilis. While, the level of CR of the xylA gene was assessed to be about l0-fold in previous work when the relative amounts of the xylA transcripts were measured in B. stearothermophilus.

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Cis-acting Replication Element Variation of the Foot-and-mouth Disease Virus is Associated with the Determination of Host Susceptibility (구제역바이러스의 숙주 특이성 결정에 연관되어있는 구제역바이러스 cis-acting replication element 변이 분석 연구)

  • Kang, Hyo Rin;Seong, Mi So;Ku, Bok Kyung;Cheong, JaeHun
    • Journal of Life Science
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    • v.30 no.11
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    • pp.947-955
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    • 2020
  • The foot-and-mouth disease virus (FMDV), a member of the Aphthovirus genus in the Picornaviridae family, affects wild and domesticated ruminants and pigs. During replication of the FMDV RNA (ribonucleic acid) genome, FMDV-encoding RNA polymerase 3D acts in a highly location-specific manner. This suggests that specific RNA structures recognized by 3D polymerase within non-coding regions of the FMDV genome assist with binding during replication. One such region is the cis-acting replication element (CRE), which functions as a template for RNA replication. The FMDV CRE adopts a stem-loop conformation with an extended duplex stem, supporting a novel 15-17 nucleotide loop that derives stability from base-stacking interactions, with the exact RNA nucleotide sequence of the CRE producing different RNA secondary structures. Here, we show that CRE sequences of FMDVs isolated in Korea from 2010 to 2017 exhibit A and O genotypes. Interestingly, variations in the RNA secondary structure of the Korean FMDVs are consistent with the phylogenetic relationships between these viruses and reveal the specificity of FMDV infections for particular host species. Therefore, we conclude that each genetic clade of Korean FMDV is characterized by a unique functional CRE and that the evolutionary success of new genetic lineages may be associated with the invention of a novel CRE motif. Therefore, we propose that the specific RNA structure of a CRE is an additional criterion for FMDV classification dependent on the host species. These findings will help correctly analyze CRE sequences and indicate the specificity of host species for future FMDV epidemics.

Effects of the Coptidis Rhizoma Extract on the Membranous Nephropathy induced by Cationic Bovine Serum Albumin in Mice (황연(黃連)이 Cationic Bovine Serum Albumin 투여로 유발된 Membranous Nephropathy Mouse Model에 미치는 영향)

  • Chae, Eun-Young;Cho, Chung-Sik;Kim, Cheol-Jung
    • The Korea Journal of Herbology
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    • v.24 no.1
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    • pp.99-110
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    • 2009
  • Objectives : The current treatment regimens for patients with nephrotic syndrome due to membranous nephropathy(MN) are based on steroids or immunosuppressive therapy with the aim of reducing proteinuria and improving outcome. Although these treatments attenuate the deterioration of renal function in MN patients, it has been suggested that all are burdened by significant toxicity. Therefore, more specific and less toxic therapies are needed. This study was to evaluate the effects of Coptidis Rhizoma Extract(CRE) on the MN induced by cBSA in mice. Methods : Mice were divided into 4 groups. One group named for 'Normal' was injected with a saline solution not to be immunized. The rest groups were treated as follows; After mice were immunized with 0.2 mg of cBSA and Freund's complete adjuvant one time every two weeks for 6 weeks, they received intra-peritoneal injection of 10 mg/kg of cBSA daily for 4 weeks. Also, they were divided into 3 groups. The first named for 'Control' was not given CRE. The second for 'CRE-250' was given oral administration of 250 mg/kg of CRE daily for 4 weeks. The third for 'CRE-500' was given 500 mg/kg of CRE. All of mice were sacrificed 4 weeks after the first immunization. We measured a body weight and 24hrs proteinuria as well as serological analysis. The morphologic changes of renal glomeruli were also observed with a light microscope and an electron microscope. Results : The levels of 24 hrs proteinuria, triglyceride, IgG, IL-6 were significantly decreased in both CRE groups. And the level of IgM was significantly decreased in CRE-250 group. In histological findings of kidney tissue, thickening of GBM and deposition of electron-density were consideraly decreased in both CRE groups. Conclusions : The present study suggests that CRE is highly effective when treating mice with MN induced by cBSA. More clinical data and studies are to be done for efficient application.

Effects of self-ligating brackets and other factors influencing orthodontic treatment outcomes: A prospective cohort study

  • Jung, Min-Ho
    • The korean journal of orthodontics
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    • v.51 no.6
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    • pp.397-406
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    • 2021
  • Objective: The purpose of this study was to evaluate the effects of self-ligating brackets (SBs) and other factors that influence orthodontic treatment outcomes. Methods: This two-armed cohort study included consecutively treated patients in a private practice. The patients were asked to choose between SBs and conventional brackets (CBs); if any patient did not have a preference, he or she was randomly allocated to the CB or SB group. All patients were treated using an identical archwire sequence. Evaluated parameters were as follows: treatment duration, number of bracket failures, poor oral hygiene, poor elastic wear, extraction, use of orthodontic mini-implants (OMI), OMI failure, American Board of Orthodontics (ABO) Discrepancy Index (DI), arch length discrepancy, and ABO Cast-Radiograph Evaluation (CRE) score. Stepwise regression analysis was performed to generate the equation for prediction of the CRE. Results: The final sample comprised 134 patients with an average age of 22.73 years. The average DI, CRE, and treatment duration were 21.81, 14.25, and 28.63 months, respectively. Analysis of covariance showed a significant difference in CRE between the CB and SB groups after adjusting for the effects of confounding variables. Stepwise regression analysis using four variables, namely extraction, SB use, poor elastic wear, and additional appliance use, could explain only 25.2% of the variance in the CRE. Conclusions: Although the CRE was significantly better for CBs than for SBs, the clinical significance of this result seems to be limited. Extraction, SB use, poor elastic wear, and additional appliance use may have significant effects on treatment outcomes.

Factors Affecting Performance of the Infection Control of Carbapenem Resistant Enterobacteriaceae of Nursing Staffs in Long-term Care Hospitals (요양병원 간호인력의 카바페넴 내성 장내세균속균종 감염관리 수행 정도에 미치는 요인)

  • Kim, Gyuri;Lee, Jong-Eun
    • Journal of Home Health Care Nursing
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    • v.29 no.2
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    • pp.204-215
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    • 2022
  • Purpose: This study aimed to identify factors affecting the carbapenem-resistant enterobacteriaceae (CRE) infection control performance of nursing staff, who closely contact patients with CRE in long-term care hospitals. Methods: A cross-sectional study design was used. A total of 135 nursing staffs working in seven long-term care hospitals in the southern and northern areas of the K province in Korea were included. We measured the CRE infection control general characteristics, knowledge, perception, and performance. Results: The main factors affecting the CRE infection control performance were education, knowledge, and perception. The model explained the 60.8% total variance in CRE infection control. Conclusion: Appropriate infection control strategies should be prepared to provide high quality nursing care and prevent the spread of CRE infection in long-term care hospitals. Establishing an efficient infection control system in long-term care hospitals is necessary.

Deletion Timing of Cic Alleles during Hematopoiesis Determines the Degree of Peripheral CD4+ T Cell Activation and Proliferation

  • Guk-Yeol Park;Gil-Woo Lee;Soeun Kim;Hyebeen Hong;Jong Seok Park;Jae-Ho Cho;Yoontae Lee
    • IMMUNE NETWORK
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    • v.20 no.5
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    • pp.43.1-43.11
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    • 2020
  • Capicua (CIC) is a transcriptional repressor that regulates several developmental processes. CIC deficiency results in lymphoproliferative autoimmunity accompanied by expansion of CD44hiCD62Llo effector/memory and follicular Th cell populations. Deletion of Cic alleles in hematopoietic stem cells (Vav1-Cre-mediated knockout of Cic) causes more severe autoimmunity than that caused by the knockout of Cic in CD4+CD8+ double positive thymocytes (Cd4-Cre-mediated knockout of Cic). In this study, we compared splenic CD4+ T cell activation and proliferation between whole immune cell-specific Cic-null (Cicf/f;Vav1-Cre) and T cell-specific Cic-null (Cicf/f;Cd4-Cre) mice. Hyperactivation and hyperproliferation of CD4+ T cells were more apparent in Cicf/f;Vav1-Cre mice than in Cicf/f;Cd4-Cre mice. Cicf/f;Vav1-Cre CD4+ T cells more rapidly proliferated and secreted larger amounts of IL-2 upon TCR stimulation than did Cicf/f;Cd4-Cre CD4+ T cells, while the TCR stimulation-induced activation of the TCR signaling cascade and calcium flux were comparable between them. Mixed wild-type and Cicf/f;Vav1-Cre bone marrow chimeras also exhibited more apparent hyperactivation and hyperproliferation of Cic-deficient CD4+ T cells than did mixed wild-type and Cicf/f;Cd4-Cre bone marrow chimeras. Taken together, our data demonstrate that CIC deficiency at the beginning of T cell development endows peripheral CD4+ T cells with enhanced T cell activation and proliferative capability.