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Glucuronidation is an enzymatic process that terminally inactivates steroid hormones, including estrogens and androgens, thereby influencing carcinogenesis in hormone-dependent cancers. While estrogens drive breast carcinogenesis via the estrogen receptor alpha (ERα), androgens play a critical role as prohormones for estrogen biosynthesis and ligands for the androgen receptor (AR). In this study, the expression and regulation of two androgen-inactivating enzymes, the UDP-glucuronosyltransferases UGT2B15 and UGT2B17, was assessed in breast cancer. In large clinical cohorts, high UGT2B15 and UGT2B17 levels positively influenced disease-specific survival in distinct molecular subgroups. Expression of these genes was highest in cases positive for ERα. In cell line models, ERα, AR, and the transcription factor FOXA1 cooperated to increase transcription via tandem binding events at their proximal promoters. ERα activity was dependent on FOXA1, facilitated by AR activation, and potently stimulated by estradiol as well as estrogenic metabolites of 5α-dihydrotestosterone. AR activity was mediated via binding to an estrogen receptor half-site 3′ to the FOXA1 and ERα-binding sites. Although AR and FOXA1 bound the UGT promoters in AR-positive/ERα-negative breast cancer cell lines, androgen treatment did not influence basal transcription levels. Ex vivo culture of human breast tissue and ERα+ tumors provided evidence for upregulation of UGT2B15 and UGT2B17 by estrogen or androgen treatment. ERα binding was evident at the promoters of these genes in a small cohort of primary tumors and distant metastases. Collectively, these data provide insight into sex steroid receptor-mediated regulation of androgen-inactivating enzymes in ERα+ breast cancer, which may have subtype-specific consequences for disease progression and outcomes. Cancer Res; 76(19); 5881–93. ©2016 AACR.

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Supplementary Figure 1. Correlation plots depicting the relationship between UGT2B15 and UGT2B17 mRNA expression in breast cancer. Supplementary Figure 3. Steroid-mediated regulation of GREB1, UGTB215 and UGTB17 in MCF7 cells. Supplementary Figure 4. Effect of ERα, FOXA1 or AR knockdown on UGT2B15 and UGT2B17 mRNA expression in MCF7 cells. Supplementary Figure 5. Genome browser images of showing FOXA1, ERα and AR chromatin occupancy at the UGT2B15 and UGT2B17 promoters in ZR75-1 and MCF7 cells. Supplementary Figure 6. Regulation of UGT2B15 and UGT2B17 mRNA expression by the AR agonist Enobosarm in MCF7 cells (A and B) and steroid-mediated regulation of UGT2B15 and UGT2B17 mRNA expression in normal breast tissues (C).

Supplementary Figure 2. Kaplan-Meier survival plots showing relationship between UGTB215 and UGTB17 levels and outcome in distinct breast cancer subtypes.

Vancomycin is widely used for treatment of infection caused by methicillin-resistant Staphylococcus aureus (MRSA) leading to an increasing appearance of low-level vancomycin-resistant isolates called heterogeneous vancomycin-intermediate S. aureus (hVISA). The mechanism of vancomycin tolerance in hVISA is still unclear. This study aimed to investigate the fatty acid compositions of S. aureus isolates under the stress environment with vancomycin. The different responses of hVISA and vancomycin-susceptible S. aureus (VSSA) may lead to more understanding the mechanism. The bacterial lipid profiles were tested three times from three extractions of each isolate cultured on tryptic soy agar (TSA) and TSA with vancomycin. Of the 30 MRSA isolates studied, 13, 12, and 5 isolates were VSSA, hVISA, and VISA, respectively. The analysis of bacterial lipid profiles showed that under vancomycin stress, there was a reduction of straight chain fatty acids (SCFAs) in VSSA isolates but an increase in branched chain fatty acids (BCFAs). In contrast, the hVISA group exhibited an increase only in the BCFAs but not in SCFAs. Of interest, vancomycin had no effect on either BCFAs or SCFAs of the VISA cells. This study provided information of bacterial adaptation during stress with vancomycin that may be helpful to overcome the resistant bacteria.

Myristicafragrans Houtt. (Nutmeg) is a widely known folk medicine across several parts of Asia, particularly used in antimicrobial treatment. Bacterial resistance involves the expression of efflux pump systems (chromosomal norA and mepA) in methicillin-resistant Staphylococcus aureus (MRSA). Crude extract (CE) and essential oil (EO) obtained from nutmeg were applied as efflux pump inhibitors (EPIs), thereby enhancing the antimicrobial activity of the drugs they were used in. The major substances in CE and EO, which function as EPIs, in a descending order of % peak area include elemicin, myristicin, methoxyeugenol, myristicin, and asarone. Here, we investigated whether the low amount of CE and EO used as EPIs was sufficient to sensitize MRSA killing using the antibiotic ciprofloxacin, which acts as an efflux system. Interestingly, synergy between ciprofloxacin and CE or EO revealed the most significant viability of MRSA, depending on norA and mepA, the latter being responsible for EPI function of EO. Therefore, CE and EO obtained from nutmeg can act as EPIs in combination with substances that act as efflux systems, thereby ensuring that the MRSA strain is susceptible to antibiotic treatment.

This research aimed 1) to study the components and indicators of creative problem solving of teachers in Mahasarakham municipal schools, 2) to study the current condition, desirable condition and how to enhance teachers in creative problem solving for schools under Mahasarakham minicipality, and 3) to develop teacher development program for creative problem solving for schools under Mahasarakham municipality. The results were as follows: 1. The components and indicators of creative problem solving were divided into 7 components and 15 indicators, including 1) problem finding ; consisting of 3 indicators: (1) emphasizing the problems (2) surveying data (3) identifying problems and causes, 2) idea finding ; consisting of 2 indicators: (1) brainstorming (2) problem solving, 3) solution finding ; consisting of 2 indicators: (1) setting criteria for alternative solution evaluating (2) alternative evaluation and improving approaches for problem solving, 4) making decision for the best solution ; consisting of 2 indicators: (1) choosing solution (2) expecting effects, 5) acceptance finding ; consisting of 2 indicators: (1) data searching and difficulties (2) problem solving plan, 6) implementation ; consisting of 2 indicators: (1) implementation (2) facing problems, 7) evaluate the problem ; consisting of indicators: (2) defining indicator for the evaluation (2) following up. 2. To study the current situation, desirable condition, and how to enhance teachers in creative problem solving for schools under Mahasarakham municipality. 2.1 The current implementation of creative problem solving of teachers in educational institutes under Mahasarakham municipality ; the overall level was moderate while the desirable implementation creative problem solving of teachers was very high. 2.2 The analysis of teacher development methods in creative problem solving which was sorted by descending order included training, seminar, workshop, internal supervision, and study visit. 3. The analysis results of enhancing teachers in creative problem solving for schools under Mahasarakham municipality were: 3.1 The needs for teacher development in creative problem solving for schools under Mahasarakham municipality, sorted by descending order were 1) finding alternative solutions (PNI modified = 0.634), 2) finding solutions (PNI modified = 0.629), 3) searching for problems (PNI modified = 0.616), 4) finding and acceping problems (PNI modified = 0.615), 5) solving problem (PNI modified = 0.602), 6) evaluating of problem solving (PNI modified = 0.594), and 7) making decision for choosing the best solution (PNI modified = 0.579). 3.2 The level of appropriateness of teacher development program in creative problem solving for educational institutes under Mahasarakam Municipality was at the highest level and the level of risks of teacher development program in creative problem solving for educational institutions in Mahasarakam Municipality was at the highest level.

This research aimed to: 1) study the factors and indicators for the health conditions of the students in the small-sized primary schools, 2) study the current conditions and problemsand the desired health conditions of the students in the small-sized primary schools. The research procedures included 2 phases composing of Phase 1: the study on the factors and indicators for the health conditions of the students in the small-sized primary schools, Phase 2: studying the current conditions and problemsand desired health conditions of the students in the small-sized primary schools. The instruments for collecting the data consisted of the questionnaires, Evaluation form and interview forms. The statistics for the data analysis covered the mean, standard deviation, percentage and the content analysis. The research findings revealed that: 1. The factors and indicators for the health conditions of the students in the small-sized primary schools included 4 factors within 42 indicators. 2. The current conditions and problemsand the desired health conditions of the students in the small-sized primary schools revealed that: 2.1 The current conditions and problemsin the health conditions of the students in the small-sized primary schools implied that most families were rather poor and gained few incomes ; the students did not live with parents but with grandparents or cousins brought the students to lack enough care affecting to the poor welfare ; the guardians did not promote them in studying, the students lacked caring for their physical health and obtained innutritious food enabling them to lack warmth causing rough mind, selfishness, untrusting people, being self-unconfident, engrossing games, stealing, damaging public assets, indiscipline and irresponsibility. 2.2 The desired health conditions of the students in the small-sized primary schools revealed that: the needs on the development for the health conditions of the students were in 4 aspects, containing physical health, mental health, social health and Spiritual health conditions.

Bloodstream infection (BSI) is a major cause of mortality in hospitalized patients worldwide. Staphylococcus aureus is one of the most common pathogens found in BSI. The conventional workflow is time consuming. Therefore, we developed a lateral flow immunoassay (LFIA) for rapid detection of S. aureus-protein A in positive blood culture samples. A total of 90 clinical isolates including 58 S. aureus and 32 non-S. aureus were spiked in simulated blood samples. The antigens were extracted by a simple boiling method and diluted before being tested using the developed LFIA strips. The results were readable by naked eye within 15 min. The sensitivity of the developed LFIA was 87.9% (51/58) and the specificity was 93.8% (30/32). When bacterial colonies were used in the test, the LFIA provided higher sensitivity and specificity (94.8% and 100%, respectively). The detection limit of the LFIA was 107 CFU/mL. Initial evaluation of the LFIA in 20 positive blood culture bottles from hospitals showed 95% agreement with the routine methods. The LFIA is a rapid, simple and highly sensitive method. No sophisticated equipment is required. It has potential for routine detection particularly in low resource settings, contributing an early diagnosis that facilitates effective treatment and reduces disease progression.

Colistin is the last resort for the treatment of infections with carbapenem-resistant (CR) Gram-negative bacteria particularly Acinetobacter baumannii (CRAB). Currently, both colistin-resistant and -heteroresistant A. baumannii isolates have been reported globally. We therefore investigated the colistin heteroresistance rate in 75 non-duplicate colistin-susceptible CRAB clinical isolates from a Thai university collected in 2016. Minimum inhibitory concentrations (MICs) of colistin for all isolates were determined by broth microdilution method and carbapenemase genes were detected by PCR methods. All isolates were genotyped by ERIC-PCR method and screened for colistin heteroresistance by modified population analysis profile (PAP) method. The colistin MIC range for the 75 isolates was 0.5–2 µg/mL, with MIC50 and MIC90 of 1 and 2 µg/mL, respectively. Thirty-three isolates (44%) were considered colistin-heteroresistant with subpopulations growing at 3–8 μg/mL of colistin. After three daily passages of the subpopulations on antibiotic-free medium, their colistin MICs ranged from 4 to > 32 µg/mL, with MIC50 and MIC90 of 32 and > 32 µg/mL, respectively. Eight different ERIC-PCR profiles were obtained among the 33 isolates and all carried blaOXA-23-like. The high rate of colistin heteroresistance in the CRAB isolates highlights the possibility of treatment failure of CRAB infections by colistin due to the selection of colistin-resistant subpopulations.

Methicillin-resistant staphylococci (MRS) are important antimicrobial-resistant pathogens in sepsis. Conventional blood cultures take 24–72 h. The polymerase chain reaction (PCR)-based methods give faster results (2–3 h) but need expensive thermal cyclers. We therefore developed an isothermal recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD) assay for rapid detection of MRS in spiked blood culture samples. Fifty-six clinical isolates including 38 mecA-carrying staphylococci and 18 non-mecA-carrying organisms as confirmed by PCR methods were studied. RPA primer set and probe specific for mecA gene (encoding penicillin-binding protein 2a) were designed. RPA reaction was carried out under isothermal condition (45 °C) within 20 min and read by LFD in 5 min. The RPA-LFD provided 92.1% (35/38) sensitivity for identifying MRS in positive blood culture samples, and no cross-amplification was found (100% specificity). This test failed to detect three mecA-carrying S.sciuri isolates. The detection limits of RPA-LFD method for identifying MRS were equal to those of PCR method. The RPA-LFD is simple, fast, and user-friendly. This method could detect the mecA gene directly from the positive blood culture samples without requirement for special equipment. This method would be useful for appropriate antibiotic therapy and infection control, particularly in a low-resource setting.

Staphylococcus aureus strains resistant to the last line antibiotic, vancomycin, have been of clinical concern. These include heterogeneous vancomycin-intermediate S. aureus (hVISA) and VISA. The hVISA phenotype cannot be detected by routine laboratory methods. Characterization of hVISA/VISA by new technologies is necessary to differentiate them rapidly from the vancomycin-susceptible isolates (VSSA). In this study, we developed a model for discrimination of hVISA from VSSA by using attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy combined with multivariate data analysis, displaying a phenotypic signature of the bacteria. ATR-FTIR spectra were acquired from a total of 59 clinical methicillin-resistant S. aureus (MRSA) isolates comprising 28 hVISA and 31 VSSA strains. Principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) were used to analyze 351 spectra of 39 isolates and develop a discrimination model for identifying hVISA and VSSA. The classification model, which was used for blind testing of 90 spectra from each of 10 hVISA, and 10 VSSA isolates, provided 100% sensitivity and specificity. The modeling revealed that the major discrimination between hVISA and VSSA phenotypes involved bands related to cell wall content (1087 and 1057 cm−1). This study showed that ATR-FTIR technique may be an alternative method for rapid detection of low-level vancomycin-resistant S. aureus.