Chronic adiponectin deficiency leads to Alzheimer’s disease-like cognitive impairments and pathologies through AMPK inactivation and cerebral insulin resistance in aged mice

(a) Immunoblotting analysis of IRβ in the hippocampus and frontal cortex of 18-month old wildtype and APN-KO mice. (b) Densitometric analysis of the ratio of IRβ. Mean ± S.E.M.; ***p < 0.001, n.s. statistically not significant; Scale bar: 100 μm. (JPG 30 kb

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals &lt;1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

First time in the European Parliament Elections: Central and Eastern Europe in the 2004 European Parliament Elections

Prve volitve novih članic Evropske unije v Evropski parlament so se prikazale kot globok paradoks v smislu povratnega procesa evropske integracije. Pridružitev je namreč na višji (elitni) ravni omogočila političnim strankam innjihovim vodjem tako nove priložnosti kot tudi nove polemike v povezavi s pojavom polarizacije mnenj o pomenu evropske integracije. Na nižji ravni pa je velika večina volilcev volitve v Evropski parlament prezrla ali celo namenoma ignorirala. Ta članek služi kot uvod v omenjeno problematiko. Njegov namen je, kot prvo razložiti pomanjkanje interesa za volitve v Evropski parlament, kot drugo raziskati domača politična gibanja pred volitvami in kot tretje pretehtati kakšen vpliv imajo te volitve na delovanje razširjene Evropske unije. Glede prihodnosti procesa evropske integracije je pomembna ozaveščenost, da vsestarnkarska podpora v novo pridruženih članicah ni več samoumevna. Namesto tega lahko pričakujemo vzpon vpilva evroskepticizma znotraj Evropske unijeThe first European Parliament elections in the new Member States in Central and Eastern Europe demonstrated a profound paradox in terms of being a feedback process of European integration. At the elite level, the accession tothe European Union has offered political parties and their leaders both new opportunities as well as a new set of issues with the emergence of a significant divide over the meanings of European integration. At the mass level, however, the first European Parliament elections were ignored by a vastmajority of voters. This paper serves as a systematic analysis of the subject. Our objective is three-fold: to explain a lack of interest in the polls, to examine the domestic political dynamics leading to the elections andto consider the implications of the elections for the workings of the enlarged European Union. As for the prospects for European integration, it is important to note that one may no longer assume a supportive cross-party consensus in the new Member States on the EU. Rather, popular antipathy towards the EU is expected to ris

Silencing BRE Expression in Human Umbilical Cord Perivascular (HUCPV) Progenitor Cells Accelerates Osteogenic and Chondrogenic Differentiation

<div><p>BRE is a multifunctional adapter protein involved in DNA repair, cell survival and stress response. To date, most studies of this protein have been focused in the tumor model. The role of BRE in stem cell biology has never been investigated. Therefore, we have used HUCPV progenitor cells to elucidate the function of BRE. HUCPV cells are multipotent fetal progenitor cells which possess the ability to differentiate into a multitude of mesenchymal cell lineages when chemically induced and can be more easily amplified in culture. In this study, we have established that BRE expression was normally expressed in HUCPV cells but become down-regulated when the cells were induced to differentiate. In addition, silencing <i>BRE</i> expression, using <i>BRE</i>-siRNAs, in HUCPV cells could accelerate induced chondrogenic and osteogenic differentiation. Hence, we postulated that BRE played an important role in maintaining the stemness of HUCPV cells. We used microarray analysis to examine the transcriptome of <i>BRE</i>-silenced cells. <i>BRE</i>-silencing negatively regulated <i>OCT4</i>, <i>FGF5</i> and <i>FOXO1A</i>. <i>BRE</i>-silencing also altered the expression of epigenetic genes and components of the TGF-β/BMP and FGF signaling pathways which are crucially involved in maintaining stem cell self-renewal. Comparative proteomic profiling also revealed that <i>BRE</i>-silencing resulted in decreased expressions of actin-binding proteins. In sum, we propose that BRE acts like an adaptor protein that promotes stemness and at the same time inhibits the differentiation of HUCPV cells.</p></div

HUCPV cells differentiated into osteoblasts and chondrocytes, 3 and 4 weeks after induction, respectively.

<p>(A) Alizarin red S staining and immunofluorescence staining with pro-collagen type-I antibody were used to demonstrate osteogenic differentiation by HUCPV cells maintained in osteogenic (Ost) medium and control (Ctl) media for 3 weeks. (B) Alcian blue staining and immunofluorescence staining with SOX9 antibody were used to demonstrate chondrogenic differentiation in HUCPV cells cultured in chondrogenic (Chon) and control (Ctl) media for 4 weeks. The nuclei were counterstained with DAPI. N = 3 independent experiments.</p

Comparative proteomics analysis of differentially expressed proteins in BRE silenced HUCPV cells.

<p>A representative silver stained 2-DE gel of total protein extracted from HUCPV cells that had been transfected with <i>BRE</i>-siRNA. When compared with control 2-DE gels, the ESI-MS/MS analysis identified proteins that were up- and down-regulated as a result of silencing <i>BRE</i>. Cytoskeletal binding proteins were especially affected. N = 3 independent experiments.</p