BNIP3 Plays Crucial Roles in the Differentiation and Maintenance of Epidermal Keratinocytes

Transcriptome analysis of the epidermis of Hes1−/− mouse revealed the direct relationship between Hes1 (hairy and enhancer of split-1) and BNIP3 (BCL2 and adenovirus E1B 19-kDa-interacting protein 3), a potent inducer of autophagy. Keratinocyte differentiation is going along with activation of lysosomal enzymes and organelle clearance, expecting the contribution of autophagy in this process. We found that BNIP3 was expressed in the suprabasal layer of the epidermis, where autophagosome formation is normally observed. Forced expression of BNIP3 in human primary epidermal keratinocytes (HPEKs) resulted in autophagy induction and keratinocyte differentiation, whereas knockdown of BNIP3 had the opposite effect. Intriguingly, addition of an autophagy inhibitor significantly suppressed the BNIP3-stimulated differentiation of keratinocytes, suggesting that BNIP3 plays a crucial role in keratinocyte differentiation by inducing autophagy. Furthermore, the number of dead cells increased in the human epidermal equivalent of BNIP3 knockdown keratinocytes, which suggests that BNIP3 is important for maintenance of skin epidermis. Interestingly, although UVB irradiation stimulated BNIP3 expression and cleavage of caspase3, suppression of UVB-induced BNIP3 expression led to further increase in cleaved caspase3 levels. This suggests that BNIP3 has a protective effect against UVB-induced apoptosis in keratinocytes. Overall, our data provide valuable insights into the role of BNIP3 in the differentiation and maintenance of epidermal keratinocytes

Sustainable and Resilient Neighbourhood Design

The objective of this thesis is to support initiation of resilience work at the neighbourhood scale, where “resilience” is the capability of dealing with future shocks and stresses (climate change related and non-related) and continuing to function. First, a framework for analyzing and improving neighbourhood resilience is developed. The key parts of this framework are the definition of the essential needs of the community, identification of future shocks/stresses, and the execution of a series of resilience analyses. This framework was partially applied to a case study neighbourhood, a proposed mixed use neighbourhood in Ontario. It was applied to identify ways to improve the resilience of the neighbourhood with respect to energy, and was found to be straightforward to use and effective in generating ideas. Second, the relationship between sustainability and resilience was explored, by examining the resilience potential of actions in the LEED for Neighbourhood Development (LEED-ND) sustainability rating system to a wide range of future shocks/stresses. LEED-ND was found to contribute resilience to a number of future shocks/stresses (especially energy shortage and heat waves), and had minimal conflicting qualities, but did not address resilience needs comprehensively, thus confirming the need for a neighbourhood resilience framework. Third, literature reviews were conducted to come up with a single set of resilience strategies to aid design for resilience. A design aid was developed consisting of 59 resilience strategies (e.g., avoidance, diversity, social capital) that improve “core,” “specified,” and “general” resilience. The design aid was used to find ways to improve the resilience of the case study neighbourhood to any threat, and also to heat waves specifically. It was found to be effective in generating many ideas, and relatively straightforward to use, albeit a little difficult in some parts and lengthy in process. Finally, a step-wise procedure for showing how the framework and design aid can be used together is presented.Ph.D

Sustained Improvements in Antimicrobial Therapy and Clinical Outcomes following a Pharmacist-Led Antimicrobial Stewardship Intervention: Uncontrolled Before–After Study

Our antimicrobial pharmacist-led intervention included: (a) a structured review of antibiotic prescriptions; (b) educating prescribers on antimicrobial therapy; (c) monthly reporting of department-level rates of blood sampling for culture. Daily review began in May 2018 and was discontinued after 10 months; however, the other interventions were conducted throughout the study period. This study aimed to evaluate the sustained impact of pharmacist’s interventions on antimicrobial therapy and clinical outcomes between the baseline (May–December 2017), intervention (May–December 2018), and post-intervention (May–December 2019) periods. The rate of blood culture collections before starting antipseudomonal agent therapy was significantly increased from the baseline to post-intervention periods (71% vs. 85%, p p = 0.038). Total use of antipseudomonal agents was reduced from the baseline to intervention periods and persisted during the post-intervention period (50.5 vs. 41.8 and 42.6 DDD per 1000 patient-days, p = 0.016 and p = 0.022, respectively). During the study period, there were significant reductions in the incidence of hospital-acquired Clostridioides difficile infection (1.12, 0.54, and 0.51 per 10,000 patient-days, respectively, p = 0.031) and 30-day mortality with bacteremia (19%, 18%, and 12%, respectively, p = 0.005). Our pharmacist-led interventions sustainably achieved appropriate antimicrobial therapy and improved clinical outcomes

Efficacy of educational intervention on reducing the inappropriate use of oral third-generation cephalosporins

Purpose: This study aimed to evaluate the efficacy of an educational intervention on reducing the inappropriate use of oral third-generation cephalosporins, the prevalence of resistant bacteria, and clinical outcomes. Methods: A before-after study was conducted to compare the data for 1 year before and after intervention at a Japanese university hospital. Educational intervention included lectures for all medical staff on oral antibiotics and educational meetings with each medical department. The primary outcome was the use of oral third-generation cephalosporins in inpatients as measured by the monthly median days of therapy (DOTs) per 1000 patient days. Secondary outcomes included the use of each oral antibiotic in inpatients and outpatients, proportion of β-lactamase-nonproducing ampicillin-resistant Haemophilus influenzae (BLNAR), penicillin-resistant Streptococcus pneumoniae (PRSP) and extended-spectrum β-lactamase producing Escherichia coli (ESBLEC), the incidence of hospital-acquired Clostridioides difficile infection (HA-CDI), and hospital mortality. Results: The use of oral third-generation cephalosporins in inpatients was significantly decreased after intervention [DOTs (interquartile range): 24.2 (23.5–25.1) vs. 3.7 (0.0–7.1), P < 0.001], and the value in outpatients was also decreased significantly. The use of fluoroquinolones and macrolides did not increase after intervention. The proportion of BLNAR, PRSP and ESBLEC did not change significantly during the study period. The incidence of HA-CDI was significantly decreased, and hospital mortality did not change after intervention. Conclusion: Educational intervention was effective in reducing the use of oral third-generation cephalosporins without increasing the use of broad-spectrum antibiotics and worsening clinical outcome. The prevalence of resistant bacteria did not change during the study period

Beneficial Effects of the Genus Aloe on Wound Healing, Cell Proliferation, and Differentiation of Epidermal Keratinocytes.

Aloe has been used as a folk medicine because it has several important therapeutic properties. These include wound and burn healing, and Aloe is now used in a variety of commercially available topical medications for wound healing and skin care. However, its effects on epidermal keratinocytes remain largely unclear. Our data indicated that both Aloe vera gel (AVG) and Cape aloe extract (CAE) significantly improved wound healing in human primary epidermal keratinocytes (HPEKs) and a human skin equivalent model. In addition, flow cytometry analysis revealed that cell surface expressions of β1-, α6-, β4-integrin, and E-cadherin increased in HPEKs treated with AVG and CAE. These increases may contribute to cell migration and wound healing. Treatment with Aloe also resulted in significant changes in cell-cycle progression and in increases in cell number. Aloe increased gene expression of differentiation markers in HPEKs, suggesting roles for AVG and CAE in the improvement of keratinocyte function. Furthermore, human skin epidermal equivalents developed from HPEKs with medium containing Aloe were thicker than control equivalents, indicating the effectiveness of Aloe on enhancing epidermal development. Based on these results, both AVG and CAE have benefits in wound healing and in treatment of rough skin

Behavior of Environmental Tritium at NIFS Toki Site of Japan

The levels of tritium in the atmosphere are nowadays almost only of natural origin and of the same range as before the era of the nuclear tests. In order to appraise the influence of tritium released from nuclear facilities to the environment, it is necessary to confirm the effect of tritium appearing overlapped on background tritium levels. Tritium concentrations and stable isotopes of oxygen and hydrogen in rain water, stream water and groundwater at the NIFS (National Institute for Fusion Science, Gifu prefecture, Japan) site were analyzed to understand behavior of the natural tritium in coupling with rain event. Conductivity, temperature and flow rate of the stream were monitored continuously. The range of tritium concentrations in rain for three year period was 0.09-0.78 Bq/l (average 0.37 +- 0.14 Bq/l). The tritium concentrations of stream water and groundwater were almost constant, 0.34 Bq/l and 0.25 Bq/l, respectively. The isotopic ratio of oxygen and hydrogen showed a typical seasonal pattern observed in Japan. Two component separation analysis was carried out for the stream water at the time of rain using isotopic ratio, conductivity and tritium concentration

Influence of AVG and CAE on the epidermal development in human epidermal equivalents.

<p>HPEKs were treated with AVG or CAE (10 μg/mL or 100 μg/mL). Frozen sections of skin equivalent model were then subjected to hematoxylin and eosin (HE) staining (A) or immunofluorescent staining for loricrin (LOR, green) and keratin 15 (K15, red) (B). The blue signals indicate nuclear staining. Scale bars, 20 μm. (C) The graphs indicate the mean ± SEM values for thickness of the whole epidermis or granular layer of the skin equivalent in micrometers. N = 20. **P<0.01, *P<0.05.</p

Influence of AVG and CAE on the expression of adhesion molecules in human epidermal keratinocytes.

<p>(A-C) HPEKs were treated with 10 μg/mL AVG or CAE for 2 days, and subjected to flow cytometry analysis (A), q-PCR analysis (B), and western blot analysis (C). (A) Representative histograms are shown. The graphs indicate the mean ± SEM values for median fluorescent intensity (MFI) from 6 independent experiments. **P<0.01, *P<0.05. (B) The graphs indicate the mean ± SEM values for relative expression from 4 independent experiments. (C) The extracted proteins were immunoblotted with the indicated antibodies. Graphs indicate the relative band intensities as determined by ImageJ software and plotted as the means of 3 independent experiments.</p