Non-wage benefits, corporate ownership and firm performance in post-communist economies: evidence from Ukraine

International audienceThe economic reforms in the transition economies of Central and Eastern Europe have fundamentally reshaped ownership and governance of economic production, notably through the privatization of former state-owned enterprises. These reforms were expected to transform management practices by displacing ‘cradle-to-grave’ welfare arrangements administered by state-owned enterprises. Using data drawn from two large samples of Ukrainian establishments, we investigate, in two different time points, the relationship between non-wage benefits and firm performance during the period of transition to a market economy (1994–2004). We found that non-wage benefits continued to be a critical feature of HRM practices in Ukraine during this period, and were positively associated with firm performance

The Convergence of High-Consequence Livestock and Human Pathogen Research and Development: A Paradox of Zoonotic Disease

Citation: Michelotti, J.M.; Yeh, K.B.; Beckham, T.R.; Colby, M.M.; Dasgupta, D.; Zuelke, K.A.; Olinger, G.G. The Convergence of High-Consequence Livestock and Human Pathogen Research and Development: A Paradox of Zoonotic Disease. Trop. Med. Infect. Dis. 2018, 3, 55.The World Health Organization (WHO) estimates that zoonotic diseases transmitted from animals to humans account for 75 percent of new and emerging infectious diseases. Globally, high-consequence pathogens that impact livestock and have the potential for human transmission create research paradoxes and operational challenges for the high-containment laboratories that conduct work with them. These specialized facilities are required for conducting all phases of research on high-consequence pathogens (basic, applied, and translational) with an emphasis on both the generation of fundamental knowledge and product development. To achieve this research mission, a highly-trained workforce is required and flexible operational methods are needed. In addition, working with certain pathogens requires compliance with regulations such as the Centers for Disease Control (CDC) and the U.S. Department of Agriculture (USDA) Select Agent regulations, which adds to the operational burden. The vast experience from the existing studies at Plum Island Animal Disease Center, other U.S. laboratories, and those in Europe and Australia with biosafety level 4 (BSL-4) facilities designed for large animals, clearly demonstrates the valuable contribution this capability brings to the efforts to detect, prepare, prevent and respond to livestock and potential zoonotic threats. To raise awareness of these challenges, which include biosafety and biosecurity issues, we held a workshop at the 2018 American Society for Microbiology (ASM) Biothreats conference to further discuss the topic with invited experts and audience participants. The workshop covered the subjects of research funding and metrics, economic sustainment of drug and vaccine development pipelines, workforce turnover, and the challenges of maintaining operational readiness of high containment laboratories

Characterization of BCE-1, a Transcriptional Enhancer Regulated by Prolactin and Extracellular Matrix and Modulated by the State of Histone Acetylation

We have previously described a 160-bp enhancer (BCE-1) in the bovine β-casein gene that is activated in the presence of prolactin and extracellular matrix (ECM). Here we report the characterization of the enhancer by deletion and site-directed mutagenesis, electrophoretic mobility shift analysis, and in vivo footprinting. Two essential regions were identified by analysis of mutant constructions: one binds C/EBP-β and the other binds MGF/STAT5 and an as-yet-unidentified binding protein. However, no qualitative or quantitative differences in the binding of these proteins were observed in electrophoretic mobility shift analysis using nuclear extracts derived from cells cultured in the presence or absence of ECM with or without prolactin, indicating that prolactin- and ECM-induced transcription was not dependent on the availability of these factors in the functional cell lines employed. An in vivo footprinting analysis of the factors bound to nuclear chromatin in the presence or absence of ECM and/or prolactin found no differences in the binding of C/EBP-β but did not provide definitive results for the other factors. Neither ECM nor prolactin activated BCE-1 in transient transfections, suggesting that the chromosomal structure of the integrated template may be required for ECM-induced transcription. Further evidence is that treatment of cells with inhibitors of histone deacetylase was sufficient to induce transcription of integrated BCE-1 in the absence of ECM. Together, these results suggest that the ECM induces a complex interaction between the enhancer-bound transcription factors, the basal transcriptional machinery, and a chromosomally integrated template responsive to the acetylation state of the histones

Testing therapeutics in cell-based assays: Factors that influence the apparent potency of drugs.

Identifying effective antivirals for treating Ebola virus disease (EVD) and minimizing transmission of such disease is critical. A variety of cell-based assays have been developed for evaluating compounds for activity against Ebola virus. However, very few reports discuss the variable assay conditions that can affect the results obtained from these drug screens. Here, we describe variable conditions tested during the development of our cell-based drug screen assays designed to identify compounds with anti-Ebola virus activity using established cell lines and human primary cells. The effect of multiple assay readouts and variable assay conditions, including virus input, time of infection, and the cell passage number, were compared, and the impact on the effective concentration for 50% and/ or 90% inhibition (EC50, EC90) was evaluated using the FDA-approved compound, toremifene citrate. In these studies, we show that altering cell-based assay conditions can have an impact on apparent drug potency as measured by the EC50. These results further support the importance of developing standard operating procedures for generating reliable and reproducible in vitro data sets for potential antivirals

Flow chart of the steps of the EBOV drug screen assay.

<p>Cells and media are prepared in 100 μl/well cell plates and incubated overnight. Drugs in 50 μl/well are transferred from drug dilution plates to cell plates using a 96-well manual benchtop pipettor for 1 h of contact. In the biosafety level-4 (BSL4) laboratory, EBOV in 50 μl/ well is transferred to the cell/drug plates using a 96-well manual benchtop pipettor for a final volume of 200 μl/well. At specific assay endpoints, cells are fixed and transferred to the BSL-2. Immunostaining was performed with a EBOV-specific antibody against VP40 and a fluorescent or chemiluminescent secondary antibody using a plate washer/Dispenser. Fluorescence is quantified on a plate reader. The HCI system (Operetta) is used to detect EBOV-positive cells and count cells with a nuclei stain (Hoechst 33342). In parallel, cytotoxicity assays (CellTiter Glo) with mock infected cells are performed at BSL-2. Luminescence is read on the Infinite^® M1000 Tecan plate reader. Data are analyzed using GraphPad Prism and/or Columbus software (Operetta).</p

Anti-EBOV activity of toremifene citrate in Vero E6 and Huh 7 cells under different conditions.

<p>(A) Vero E6 cells and (B) Huh 7 cells were infected at varying MOIs with different assay end points and treated with toremifene citrate. EC₅₀s were determined from 8-point dose response curves using the fluorescent assay. (C, D) EC₅₀s of toremifene citrate with corresponding assay end points or MOIs are shown for comparison. Representative graphs from 1 to 4 independent experiments are shown. Abbreviations: EBOV, Ebola virus; EC₅₀, half maximal effective concentration; h, hour; MOI, multiplicity of infection; N.A., not applicable.</p

Impact of exposure time and virus input on efficacy of toremifene citrate.

<p>(A) Huh 7 cells were infected at an MOI of 1, and antiviral activity of toremifene citrate was evaluated at indicated time points. (B) Huh 7 cells were infected at indicated MOIs and antiviral activity of toremifene citrate was evaluated at 72 hpi. (C, D) EC₅₀s with corresponding assay endpoints or MOIs are shown for comparison. The experiment was performed twice using the fluorescent assay. Representative graphs are shown. Abbreviations:EC₅₀, half maximal effective concentration; hpi, hours post-inoculation; MOI, multiplicity of infection.</p

Effect of virus input and assay endpoint on the performance of EBOV drug screen assays.

<p>Effect of virus input and assay endpoint on the performance of EBOV drug screen assays.</p

Impact of cell passaging on EBOV infection.

<p>Vero E6 (A) and Huh 7 (B) cells were passaged and infected with EBOV/Mak at an MOI of 1 at 48 hpi, the plates were fixed, stained, and the percentage of EBOV-positive cells was determined by HCI. Each point is the mean of 3 replicate wells and represents an independent experiment. For each passage, the median value of all experiments was determined. Abbreviations: EBOV, Ebola virus; HCI, high content imaging; hpi, hours post-inoculation; Mak, Makona.</p