Discovery of benzimidazole-indazole derivatives as potent FLT3-tyrosine kinase domain mutant kinase inhibitors for acute myeloid leukemia.

The FMS-like tyrosine kinase 3 (FLT3) gene encodes a class III receptor tyrosine kinase that is expressed in hematopoietic stem cells. The mutations of FLT3 gene found in 30% of acute myeloid leukemia (AML), leads to an abnormal constitutive activation of FLT3 kinase of the receptor and results in immature myeloblast cell proliferation. Although small molecule drugs targeting the FLT3 kinase have been approved, new FLT3 inhibitors are needed owing to the side effects and drug resistances arising from kinase domain mutations, such as D835Y and F691L. In this study, we have developed benzimidazole-indazole based novel inhibitors targeting mutant FLT3 kinases through the optimization of diverse chemical moieties substituted around the core skeleton. The most optimized compound 22f exhibited potent inhibitory activities against FLT3 and FLT3/D835Y, with IC50 values of 0.941 and 0.199 nM, respectively. Furthermore, 22f exhibited strong antiproliferative activity against an AML cell line, MV4-11 cells with a GI50 of 0.26 nM. More importantly, 22f showed single-digit nanomolar GI50 values in the mutant FLT kinase expressed Ba/F3 cell lines including FLT-D835Y (GI50 = 0.29 nM) and FLT3-F691L (GI50 = 2.87 nM). Molecular docking studies indicated that the compound exhibits a well-fitted binding mode as a type 1 inhibitor in the homology model of active conformation of FLT3 kinase.

Risk factors for Pneumocystis pneumonia with acute respiratory failure among kidney transplant recipients.

PURPOSE: One of the rare life-threatening fungal infections is pneumocystis pneumonia (PCP). Immunocompromised patients are the main vulnerable population. We investigate the risk factors associated with the development of severe PCP infection with acute respiratory failure after kidney transplantation. MATERIALS AND METHODS: This is a retrospective, single-center, case-control study. PCP patients who are kidney transplant recipients and required high-flow oxygen support or mechanical ventilation between March 2009 and February 2017 were included in the study. The comparison was conducted between the non-severe and severe PCP groups. To identify associated risk factors, we performed univariate and multivariate logistic regression. RESULTS: Among the total 2,330 kidney transplant recipients, 50 patients (2.1%) were diagnosed with PCP. Of these, 27 patients (54.0%) had severe PCP and 7 patients (14.0%) died, all of them were severe PCP patients. In the severe PCP group, the time from transplantation to PCP diagnosis (23.4 ± 24.9 months vs. 13.7 ± 9.9 months, p = 0.090) was insignificantly faster than in the non-severe PCP group. According to multiple logistic regression analysis, the significant risk factors associated with severe PCP were as follows, age (odds ratios (OR) 1.07; 95% confidence intervals (CI): 1.01-1.13; p = 0.027), time from transplantation to PCP diagnosis (odds ratios (OR) 0.92; 95% confidence intervals (CI): 0.86-0.99; p = 0.024), lymphopenia (OR 6.48; 95% CI: 1.05-40.09; p = 0.044), and history of acute rejection within 1 year (OR 8.28; 95% CI: 1.29-53.20; p = 0.026). CONCLUSION: Patients who have lymphopenia at the time of hospital admission or have been recently treated with acute rejection are more likely to progress to severe PCP, requiring intensive monitoring and aggressive treatment.

A Novel Muscle Atrophy Mechanism: Myocyte Degeneration Due to Intracellular Iron Deprivation.

Muscle atrophy is defined as the progressive degeneration or shrinkage of myocytes and is triggered by factors such as aging, cancer, injury, inflammation, and immobilization. Considering the total amount of body iron stores and its crucial role in skeletal muscle, myocytes may have their own iron regulation mechanism. Although the detrimental effects of iron overload or iron deficiency on muscle function have been studied, the molecular mechanism of iron-dependent muscle atrophy has not been elucidated. Using human muscle tissues and in the mouse rotator cuff tear model, we confirmed an association between injury-induced iron depletion in myocytes and muscle atrophy. In differentiated C2C12 myotubes, the effects of iron deficiency on myocytes and the molecular mechanism of muscle atrophy by iron deficiency were evaluated. Our study revealed that the lower iron concentration in injured muscle was associated with the upregulation of ferroportin, an iron exporter that transports iron out of cells. Ferroportin expression was increased by hypoxia-inducible factor 1α (HIF1α), which is activated by muscle injury, and its expression is controlled by HIF1 inhibitor treatment. Iron deprivation caused myocyte loss and a marked depletion of mitochondrial membrane potential leading to muscle atrophy, together with increased levels of myostatin, the upstream regulator of atrogin1 and muscle RING-finger protein-1 (MuRF1). Myostatin expression under iron deficiency was mediated by an orphan nuclear receptor, dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome (DAX1).

Differential regulation of the water channel protein aquaporins in chondrocytes of human knee articular cartilage by aging.

Knee cartilage is in an aqueous environment filled with synovial fluid consisting of water, various nutrients, and ions to maintain chondrocyte homeostasis. Aquaporins (AQPs) are water channel proteins that play an important role in water exchange in cells, and AQP1, -3, and -4 are known to be expressed predominantly in cartilage. We evaluated the changes in AQP expression in chondrocytes from human knee articular cartilage in patients of different ages and identified the key factor(s) that mediate age-induced alteration in AQP expression. The mRNA and protein expression of AQP1, -3 and -4 were significantly decreased in fibrocartilage compared to hyaline cartilage and in articular cartilage from older osteoarthritis patients compared to that from young patients. Gene and protein expression of AQP1, -3 and -4 were altered during the chondrogenic differentiation of C3H10T1/2 cells. The causative factors for age-associated decrease in AQP included H2O2, TNFα, and HMGB1 for AQP1, -3, and -4, respectively. In particular, the protective effect of AQP4 reduction following HMGB1 neutralization was noteworthy. The identification of other potent molecules that regulate AQP expression represents a promising therapeutic approach to suppress cartilage degeneration during aging.

Two-Year Hospital-Wide Surveillance of Central Line-Associated Bloodstream Infections in a Korean Hospital.

BACKGROUND: Surveillance and interventions of central line-associated bloodstream infections (CLABSIs) had mainly been targeted in intensive care units (ICUs). Central lines are increasingly used outside ICUs. Therefore, we performed a hospital-wide survey of CLABSIs to evaluate the current status and develop strategies to reduce CLBASI rates. METHODS: All hospitalized patients with central venous catheters (CVCs) were screened for CLABSIs from January 2014 through December 2015 at a 1,328 bed tertiary care teaching hospital in Korea using an electronic data-collecting system. Clinical information including type of CVC was collected. CLABSI rates were calculated using the definitions of the National Health and Safety Network after excluding mucosal barrier injury laboratory-confirmed bloodstream infection (BSI). RESULTS: A total of 154 CLABSIs were identified, of which 72 (46.8%) occurred in general wards and 82 (53.2%) in ICUs (0.81 and 2.71 per 1,000 catheter days), respectively. Non-tunneled CVCs were most common (68.6%) among 70 CLABSI events diagnosed within one week of their maintenance. On the other hand, tunneled CVCs and peripherally inserted central catheters (PICCs) were more common (60.5%) among 114 CLABSI events diagnosed more than a week after maintenance. Whereas the majority (72.2%) of CLABSIs in ICUs were associated with non-tunneled CVCs, tunneled CVCs (38.9%) and PICCs (36.8%) were more common in general wards. CONCLUSION: CLABSIs are less common in general wards than in ICUs, but they are more often associated with long-term indwelling catheters. Therefore, interventions to prevent CLABSIs should be tailored according to the type of ward and type of catheter.

Impact of a hand hygiene campaign in a tertiary hospital in South Korea on the rate of hospital-onset methicillin-resistant Staphylococcus aureus bacteremia and economic evaluation of the campaign.

BACKGROUND: Hand hygiene (HH) is the most important factor affecting health care-associated infections. METHODS: We introduced a World Health Organization HH campaign in October 2010. The monthly procurement of hand sanitizers per 1,000 patient days was calculated, and the monthly incidence of methicillin-resistant Staphylococcus aureus bacteremia (MRSAB), classified into community- and hospital-onset (HO), was measured from a microbiologic laboratory database. Trends of MRSAB incidence were assessed using Bayesian structural time series models. A cost-benefit analysis was also performed based on the economic burden of HO MRSAB in Korea. RESULTS: Procurement of hand sanitizers increased 134% after the intervention (95% confidence interval [CI], 120%-149%), compared with the preintervention period (January 2008-September 2010). In the same manner, HH compliance improved from 33.2% in September 2010 to 92.2% after the intervention. The incidence of HO MRSAB per 100,000 patient days decreased 33% (95% CI, -57% to -7.8%) after the intervention. Because there was a calculated reduction of 65 HO MRSAB cases during the intervention period, the benefit outweighed the cost (total benefit [$851,565]/total cost [$167,495] = 5.08). CONCLUSIONS: Implementation of the HH campaign led to increased compliance and significantly reduced HO MRSAB incidence; it was also cost saving.

The education and practice program for medical students with quantitative and qualitative fit test for respiratory protective equipment.

Tuberculosis infection is prevalent in Korea and health care workers are vulnerable to tuberculosis infection in the hospital. The aims of this study were to develop and validate an education program that teaches senior medical students how to wear and choose the proper size and type of respiratory protective equipment (RPE), which may help reduce the risk of contracting Mycobacterium tuberculosis (MTB) from patients. Overall, 50 senior medical students participated in this education program. Methods of choosing the proper type of RPE, performing a fit check of the RPE, and choosing a suitable mask size were taught by certified instructors using the real-time quantitative fit test (QNFT). The validity of education program was evaluated with qualitative fit test (QLFT) before and after the education as pass or fail. The education program was effective, as shown by the significantly pass rate (increased 30 to 74%) in the QLFT after the education program (p<0.05). Among study participants, changing mask size from medium to small significantly increased the pass rate (p<0.001). Incorporation of this program into the medical school curriculum may help reduce risk of MTB infection in medical students working in the hospital.

Assessing real-time performances of N95 respirators for health care workers by simulated workplace protection factors.

To assess performances of N95 respirators for Health Care Workers (HCWs) in a simulated health-care setting, we measured the Simulated Workplace Protection Factors (SWPFs) in real-time from the volunteers. A total of 49 study subjects, wearing 3 M respirator Model N95 1860 and 1860S, were fit tested using the OSHA Exercise Regimen. The test subjects were asked to perform simulated scenarios, including patient assessments, suction, and intravenous injection (IV) treatment. Two TSI PortaCount instruments continuously measured concentrations in the respirator and the room concentration. For Quantitative Fit Testing (QNFT), 36 out of 49 (73.5%) passed the fit factor (FF) criteria set at 100 and 13 (26.5%) failed. The results of QNFT were found to have a low correlation with SWPF, with R(2)=0.32. The geometric means (GM) and geometric standard deviations (GSD) of SWPF were 68.8 (1.1) for those subjects who passed and 39.6 (1.3) for those who failed. Real-time assessments of SWPF showed that lower SWPFs were; moving head up and down, and bending at the waist. This study identifies the needs for providing different sizes of respirators for HCWs and the importance of performing fit tests for HCWs regularly. And particular movements were identified as attributing factors affecting more on SWPFs.

Successful containment of carbapenem-resistant Enterobacteriaceae by strict contact precautions without active surveillance.

BACKGROUND: Carbapenem-resistant Enterobacteriaceae (CRE) are a growing problem worldwide. Guidelines focus on carbapenemase-producing organisms, and little is known about whether strict adherence to infection control measures is effective for CRE without carbapenemase. During 2009, CRE increased markedly in a tertiary hospital, and enhanced infection control measures without active surveillance were adopted. METHODS: Beginning in April 2010, enhanced antimicrobial stewardship, strict contact precautions, and cohort isolation were adopted. After September 2010, hand hygiene performance was prospectively monitored by active surveillance, and results were monthly fed back to medical personnel. Available carbapenem-resistant Escherichia coli (ECO) and carbapenem-resistant Klebsiella pneumoniae (KPN) isolated during 2008-2010 were characterized. Imipenem and meropenem minimal inhibitory concentrations were confirmed by E-test (AB biodisk, Solna, Sweden). Phenotypic screening assays and polymerase chain reaction (PCR) amplification of known ß-lactamase and carbapenemase genes were performed. RESULTS: From 3,511 ECO and 2,279 KPN, 44 (0.76%) were CRE (3 ECO, 41 KPN). CRE incidence rates rose from 1.61 in 2008 to 5.49 in 2009; they rose further to 9.81 per 100,000 patient days in early 2010. After adoption of strict infection control measures, CRE frequency fell back in 2011 and remained at baseline afterward. Phenotypic screening and PCR showed AmpC ß-lactamase and extended spectrum ß-lactamases with or without loss of porins; carbapenemases were not detected. CONCLUSION: Enhanced infection control measures, even without active surveillance, seem effective to prevent further spread of CRE in a low-prevalence setting with mainly carbapenemase-nonproducing CRE.

Outcome of cefazolin prophylaxis for total knee arthroplasty at an institution with high prevalence of methicillin-resistant Staphylococcus aureus infection.

OBJECTIVES: The aim of this study was to evaluate the outcome of cefazolin prophylaxis for total knee arthroplasty (TKA) in a hospital with a high prevalence of methicillin-resistant Staphylococcus aureus (MRSA) infection. METHODS: Since July 1, 2006, we have applied a 'care bundle' to TKA to prevent surgical site infection (SSI) without using vancomycin as antimicrobial prophylaxis, in accordance with the 1999 Hospital Infection Control Practices Advisory Committee guidelines. All patients undergoing TKA from July 1, 2006 to September 30, 2009 were enrolled. We reviewed data on SSI collected prospectively as part of routine infection control surveillance. RESULTS: Of 1323 TKAs, an SSI developed in 14 (1.06%) cases, which is comparable to the percentage obtained in other previous reports. When stratified by the National Nosocomial Infection Surveillance risk index, SSI rates were 0.86% (8/926), 1.30% (5/384), and 7.69% (1/13) in risk categories 0, 1, and 2, respectively. Of 14 SSIs, four (29%) were classified as superficial incisional, two (14%) as deep incisional, and eight (57%) as organ-space SSI. CONCLUSIONS: Our data suggest that antimicrobial prophylaxis using only cefazolin can maintain low SSI rates if other important infection management measures are employed, even where there is a high prevalence of MRSA infection.

Effect of storage in juice with or without pulp and/or calcium lactate on the subsequent survival of Escherichia coli O157:H7 in simulated gastric fluid.

A study was conducted to evaluate the effect of storing Escherichia coli O157:H7 in fruit or vegetable juices with or without pulp and/or calcium lactate, on the bacterial resistance to a simulated gastric fluid (SGF, pH 1.5). Apple, carrot, orange, and tomato juices containing pulp or freed from pulp by filtration were used in this study. Calcium lactate at about 1.4 g/l was added to juices to obtain calcium supplemented juices. Juices with or without pulp and/or calcium lactate were inoculated with E. coli O157:H7 and then were stored at 7 degrees C for 0, 1, 2, or 4 days. The acid resistance of cells stored in juices with or without pulp and/or calcium lactate was determined by incubating in SGF for 90 or 240 min at 37 degrees C. Cells stored in apple juice for 4 days, carrot juice for 2 days, and orange juice for 4 days with pulp only had greater acid resistance, while all cells stored in tomato juice with pulp had greater acid resistance than cells stored in juice without pulp. The D-values of cells stored in supplemented apple and orange juices with calcium lactate declined 1.7-3.5 fold, whereas D-values of cells stored in supplemented tomato juice decreased by about 1.4-fold when compared to cells stored in juice without calcium lactate after exposure in SGF. These results indicate that storing E. coli O157:H7 in juices with pulp had little or no effect on the acid resistance of cells during subsequent exposure in SGF. Calcium lactate supplemented into juices could dramatically decrease the ability of E. coli O157:H7 to survive in SGF, possibly reducing the risk of foodborne illness by juice products.