The human ankyrin 1 promoter insulator sustains gene expression in a β-globin lentiviral vector in hematopoietic stem cells.

Lentiviral vectors designed for the treatment of the hemoglobinopathies require the inclusion of regulatory and strong enhancer elements to achieve sufficient expression of the β-globin transgene. Despite the inclusion of these elements, the efficacy of these vectors may be limited by transgene silencing due to the genomic environment surrounding the integration site. Barrier insulators can be used to give more consistent expression and resist silencing even with lower vector copies. Here, the barrier activity of an insulator element from the human ankyrin-1 gene was analyzed in a lentiviral vector carrying an antisickling human β-globin gene. Inclusion of a single copy of the Ankyrin insulator did not affect viral titer, and improved the consistency of expression from the vector in murine erythroleukemia cells. The presence of the Ankyrin insulator element did not change transgene expression in human hematopoietic cells in short-term erythroid culture or in vivo in primary murine transplants. However, analysis in secondary recipients showed that the lentiviral vector with the Ankyrin element preserved transgene expression, whereas expression from the vector lacking the Ankyrin insulator decreased in secondary recipients. These studies demonstrate that the Ankyrin insulator may improve long-term β-globin expression in hematopoietic stem cells for gene therapy of hemoglobinopathies

Water Footprint Assessment of a Tea Product in Sri Lanka A Case Study of Black Tea and Selected Tea Flavours

The rising global population and depleting aquifers will lead to a global water crisis indifferent perspectives. In this context business world will face physical risk, reputational risk,regulatory risk, and financial risk that may translate into increased costs and/or reducedrevenues. Presently most of the corporate sector is working towards to efficient watermanagement in their business operations to overcome the farseeing water related risks. Waterfootprint is the one of the emerging tool to assess the total impact to the water by an entity.This study emphasizes the business valve of estimation of product water footprint for a SriLankan tea product to capture the competitive advantage in global market.The aim of this study is to assess the ‘virtual water footprint’ through life cycle of a teaproduction, mainly focusing on the fresh water consumption starting from the cultivationstage of green tea leaves and continuing on to tea processing and transportation to finally thepackaging stage. The study distinguishes between three types of impact: evaporation ofinfiltrated rainwater for tea growth (green water use), withdrawal of ground- or surface waterfor irrigation or processing (blue water use) and water pollution during growth or processing.The latter impact is quantified in terms of the dilution volume necessary to assimilate thepollution (Gary water). For the study, virtual water footprints of six flavours of tea productswere analysed according to the water footprint manual published by water FootprintNetwork.According to the study, the average virtual water footprint of the 1.5 g tea bag from the greenleaf production up to the manufacturer’s gate for black tea is 19.8 liters (l), green tea is 18.46l and average virtual water footprint for flavoured tea is 20.22 l ± 7.04 l per tea bag. Averagevirtual water footprint for the ingredients including raw tea and flavours contribute 93.6% forthe total water footprint and the 6.4% of the total water footprint accountable for the virtualwater footprint of the packaging materials.

Values and Diversity of Organisational Work Ethicality: Lessons Learnt from Sri Lankan Entrepreneurs

The aim of this paper is to examine values as a component of organisational work ethicality, uncovering the contextual realities of values and explore the diversity of work ethicality in the entrepreneurial context in Sri Lanka. Organisational work ethicality is explained through egoistic work ethicality, utilitarian work ethicality and deontological work ethicality. With a view to get a deeper understanding of the diversity of organisational work ethicality, a qualitative approach is adopted. Twelve entrepreneurs were selected for in-depth interviews. A thematic analysis was carried out for field notes with the support of NVivo 6 version. Findings of this study reveal that the entrepreneurial thinking and their interpretations derived different social meanings of organisational work ethicality. The final outcome shows that rather than single dominant work ethicality, a combination of work ethicalities was preferred by most of the entrepreneurs. Within this mix of ethicalities, deontological ethicality and egoistic work ethicality seem to be equally practised by the entrepreneurs. The utilitarian ethicality seems to be practised at a minimum level. As a result, the implicational value of this study lies on at the organisational level and policy making level to rethink and reestablish a mechanism to improve ethical aspects of the businesses in order to maximise social well-being while doing the right things for the society and strengthening shareholders’ protection. KeywordsBusiness Ethics, Business Practices, Deontological Work Ethicality,Egoistic Work Ethicality, Entrepreneur, Utilitarian Work Ethicality, Value

Gas-phase Fragmentation of Deprotonated p-Hydroxyphenacyl Derivatives

Electrospray ionization of methanolic solutions of p-hydroxyphenacyl derivatives HO-C6H4-C(O)-CH2-X (X = leaving group) provides abundant signals for the deprotonated species which are assigned to the corresponding phenolate anions −O-C6H4-C(O)-CH2-X. Upon collisional activation in the gas phase, these anions inter alia undergo loss of a neutral “C8H6O2” species concomitant with formation of the corresponding anions X−. The energies required for the loss of neutral roughly correlate with the gas phase acidities of the conjugate acids (HX). Extensive theoretical studies performed for X = CF3COO in order to reveal the energetically most favorable pathway for the formation of neutral “C8H6O2” suggest three different routes of similar energy demands, involving a spirocyclopropanone, epoxide formation, and a diradical, respectively

Ion transport on self-assembled block copolymer electrolytes with different side chain chemistries

Ion-exchange membranes (IEMs) are used in electrochemical systems for a wide variety of applications, including water purification, mineral recovery, and energy storage and conversion. These materials often dictate the ohmic overpotential drop in electrochemical systems and can have a profound impact on process efficiency. Central to the rationale design of ion-conducting polymers is a fundamental understanding as to how chemical composition and macromolecular architecture influence ion and water transport. Herein, we report the preparation of three microphase separated block copolymer electrolytes (BCEs) with long-range order that have different side chain chemistries in the ionic domain. The side chain variants are alkyl and alkoxy pendants and a zwitterionic group. The side chain chemistries were installed post-assembly of crosslinked block copolymers. Unexpectedly, we observed that more hydrophobic alkyl side chain yields about an order of magnitude greater ionic conductivity when compared to alkoxy and zwitterionic side chains despite similar IEC values and water uptake. Molecular dynamics simulations reveal that the hydrophilic alkoxy moieties and zwitterion structure coordinate with water making less free water available for mediating ionic conductivity. Conversely, the hydrophobic alkyl side chains give rise to large, interconnected water clusters that promote ionic conduction of the counterion

Rho GTPase Cdc42 Is a Direct Interacting Partner of Adenomatous Polyposis Coli Protein and Can Alter Its Cellular Localization

Adenomatous Polyposis Coli (APC) is a tumor suppressor gene product involved in colon cancer. APC is a large multidomain molecule of 2843 amino acid residues and connects cell-cell adhesion, the F-actin/microtubule cytoskeleton and the nucleus. Here we show that Cdc42 interacts directly with the first three armadillo repeats of APC by yeast two-hybrid screens. We confirm the Cdc42-APC interaction using pulldown assays in vitro and FRET assays in vivo. Interestingly, Cdc42 interacts with APC at leading edge sites where F-actin is enriched. In contrast, Cdc42 interacts with the truncated mutant APC1–1638 in cellular puncta associated with the golgi-lysozome pathway in transfected CHO cells. In HCT116 and SW480 cells, Cdc42 induces the relocalization of endogenous APC and the mutant APC1–1338 to the plasma membrane and cellular puncta, respectively. Taken together, these data indicate that the Cdc42-APC interaction induces localization of both APC and mutant APC and may thus play a direct role in the functions of these proteins

Hacking into bacterial biofilms: a new therapeutic challenge

Microbiologists have extensively worked during the past decade on a particular phase of the bacterial cell cycle known as biofilm, in which single-celled individuals gather together to form a sedentary but dynamic community within a complex structure, displaying spatial and functional heterogeneity. In response to the perception of environmental signals by sensing systems, appropriate responses are triggered, leading to biofilm formation. This process involves various molecular systems that enable bacteria to identify appropriate surfaces on which to anchor themselves, to stick to those surfaces and to each other, to construct multicellular communities several hundreds of micrometers thick, and to detach from the community. The biofilm microbial community is a unique, highly competitive, and crowded environment facilitating microevolutionary processes and horizontal gene transfer between distantly related microorganisms. It is governed by social rules, based on the production and use of "public" goods, with actors and recipients. Biofilms constitute a unique shield against external aggressions, including drug treatment and immune reactions. Biofilm-associated infections in humans have therefore generated major problems for the diagnosis and treatment of diseases. Improvements in our understanding of biofilms have led to innovative research designed to interfere with this process

Lack of the Delta Subunit of RNA Polymerase Increases Virulence Related Traits of Streptococcus mutans

The delta subunit of the RNA polymerase, RpoE, maintains the transcriptional specificity in Gram-positive bacteria. Lack of RpoE results in massive changes in the transcriptome of the human dental caries pathogen Streptococcus mutans. In this study, we analyzed traits of the ΔrpoE mutant which are important for biofilm formation and interaction with oral microorganisms and human cells and performed a global phenotypic analysis of its physiological functions. The ΔrpoE mutant showed higher self-aggregation compared to the wild type and coaggregated with other oral bacteria and Candida albicans. It formed a biofilm with a different matrix structure and an altered surface attachment. The amount of the cell surface antigens I/II SpaP and the glucosyltransferase GtfB was reduced. The ΔrpoE mutant displayed significantly stronger adhesion to human extracellular matrix components, especially to fibronectin, than the wild type. Its adhesion to human epithelial cells HEp-2 was reduced, probably due to the highly aggregated cell mass. The analysis of 1248 physiological traits using phenotype microarrays showed that the ΔrpoE mutant metabolized a wider spectrum of carbon sources than the wild type and had acquired resistance to antibiotics and inhibitory compounds with various modes of action. The reduced antigenicity, increased aggregation, adherence to fibronection, broader substrate spectrum and increased resistance to antibiotics of the ΔrpoE mutant reveal the physiological potential of S. mutans and show that some of its virulence related traits are increased

Calibration and simulation of ionization signal and electronics noise in the ICARUS liquid argon time projection chamber

The ICARUS liquid argon time projection chamber (LArTPC) neutrino detector has been taking physics data since 2022 as part of the Short-Baseline Neutrino (SBN) Program. This paper details the equalization of the response to charge in the ICARUS time projection chamber (TPC), as well as data-driven tuning of the simulation of ionization charge signals and electronics noise. The equalization procedure removes non-uniformities in the ICARUS TPC response to charge in space and time. This work leverages the copious number of cosmic ray muons available to ICARUS at the surface. The ionization signal shape simulation applies a novel procedure that tunes the simulation to match what is measured in data. The end result of the equalization procedure and simulation tuning allows for a comparison of charge measurements in ICARUS between Monte Carlo simulation and data, showing good performance with minimal residual bias between the two