Multicistronic lentiviral vectors containing the FMDV 2A cleavage factor demonstrate robust expression of encoded genes at limiting MOI

BACKGROUND: A number of gene therapy applications would benefit from vectors capable of expressing multiple genes. In this study we explored the feasibility and efficiency of expressing two or three transgenes in HIV-1 based lentiviral vector. Bicistronic and tricistronic self-inactivating lentiviral vectors were constructed employing the internal ribosomal entry site (IRES) sequence of encephalomyocarditis virus (EMCV) and/or foot-and-mouth disease virus (FMDV) cleavage factor 2A. We employed enhanced green fluorescent protein (eGFP), O(6)-methylguanine-DNA-methyltransferase (MGMT), and homeobox transcription factor HOXB4 as model genes and their expression was detected by appropriate methods including fluorescence microscopy, flow cytometry, immunocytochemistry, biochemical assay, and western blotting. RESULTS: All the multigene vectors produced high titer virus and were able to simultaneously express two or three transgenes in transduced cells. However, the level of expression of individual transgenes varied depending on: the transgene itself; its position within the construct; the total number of transgenes expressed; the strategy used for multigene expression and the average copy number of pro-viral insertions. Notably, at limiting MOI, the expression of eGFP in a bicistronic vector based on 2A was ~4 times greater than that of an IRES based vector. CONCLUSION: The small and efficient 2A sequence can be used alone or in combination with an IRES for the construction of multicistronic lentiviral vectors which can express encoded transgenes at functionally relevant levels in cells containing an average of one pro-viral insert

Is Diversity the Missing Link in Coastal Fisheries Management?

Fisheries management has historically focused on the population elasticity of target fish based primarily on demographic modeling, with the key assumptions of stability in environmental conditions and static trophic relationships. The predictive capacity of this fisheries framework is poor, especially in closed systems where the benthic diversity and boundary effects are important and the stock levels are low. Here, we present a probabilistic model that couples key fish populations with a complex suite of trophic, environmental, and geomorphological factors. Using 41 years of observations we model the changes in eastern Baltic cod (Gadus morhua), herring (Clupea harengus), and Baltic sprat (Sprattus sprattus balticus) for the Baltic Sea within a Bayesian network. The model predictions are spatially explicit and show the changes of the central Baltic Sea from cod- to sprat-dominated ecology over the 41 years. This also highlights how the years 2004 to 2014 deviate in terms of the typical cod–environment relationship, with environmental factors such as salinity being less influential on cod population abundance than in previous periods. The role of macrozoobenthos abundance, biotopic rugosity, and flatfish biomass showed an increased influence in predicting cod biomass in the last decade of the study. Fisheries management that is able to accommodate shifting ecological and environmental conditions relevant to biotopic information will be more effective and realistic. Non-stationary modelling for all of the homogeneous biotope regions, while acknowledging that each has a specific ecology relevant to understanding the fish population dynamics, is essential for fisheries science and sustainable management of fish stocks

3-(2H-1,3-Benzodioxol-5-ylmeth­yl)-2-(2-meth­oxy­phen­yl)-1,3-thia­zolidin-4-one

The title mol­ecule, C18H17NO4S, features a 1,3-thia­zolidine ring that is twisted about the S—C(methyl­ene) bond. With reference to this ring, the 1,3-benzodioxole and benzene rings lie to either side and form dihedral angles of 69.72 (16) and 83.60 (14)°, respectively, with the central ring. Significant twisting in the mol­ecule is confirmed by the dihedral angle of 79.91 (13)° formed between the outer rings. Linear supra­molecular chains along the a-axis direction mediated by C—H⋯O inter­actions feature in the crystal packing

4-(Pyrimidin-2-yl)-1-thia-4-aza­spiro­[4.5]decan-3-one

The title compound, C12H15N3OS, features an envelope conformation for the 1,3-thia­zolidin-4-one ring with the S atom as the flap atom. The pyrimidine ring is almost orthogonal to the 1,3-thia­zolidin-4-one ring as indicated by the N—C—C—N torsion angle of −111.96 (18)°. Supra­molecular dimers are formed in the crystal structure through the agency of C—H⋯O contacts occurring between centrosymmetrically related mol­ecules. These are linked into supra­molecular tapes along [100] via C—H⋯S contacts

(E)-1-(2,4-Dinitro­phen­yl)-2-pentyl­idenehydrazine

The title compound, C11H14N4O4, is essentially planar with an r.m.s. deviation for the 19 non-H atoms of 0.152 Å. The conformation about the C=N bond is E, and the mol­ecule has a U-shape as the butyl group folds over towards the aromatic system. An intra­molecular C—H⋯N inter­action occurs. The crystal packing is dominated by N—H⋯O hydrogen bonding and C—H⋯O contacts, leading to twisted zigzag supra­molecular chains along the c direction. The crystal packing brings two nitro O atoms into an unusually close proximity of 2.686 (4) Å. While the nature of this inter­action is not obvious, there are several precendents for such short nitro–nitro O⋯O contacts of less than 2.70 Å in the crystallographic literature

Progress and prospects for event tourism research

This paper examines event tourism as a field of study and area of professional practice updating the previous review article published in 2008. In this substantially extended review, a deeper analysis of the field’s evolution and development is presented, charting the growth of the literature, focusing both chronologically and thematically. A framework for understanding and creating knowledge about events and tourism is presented, forming the basis which signposts established research themes and concepts and outlines future directions for research. In addition, the review article focuses on constraining and propelling forces, ontological advances, contributions from key journals, and emerging themes and issues. It also presents a roadmap for research activity in event tourism