Clinical applications of gene therapy for rare diseases: A review

Rare diseases collectively exact a high toll on society due to their sheer number and overall prevalence. Their heterogeneity, diversity, and nature pose daunting clinical challenges for both management and treatment. In this review, we discuss recent advances in clinical applications of gene therapy for rare diseases, focusing on a variety of viral and non-viral strategies. The use of adeno-associated virus (AAV) vectors is discussed in the context of Luxturna, licenced for the treatment of RPE65 deficiency in the retinal epithelium. Imlygic, a herpes virus vector licenced for the treatment of refractory metastatic melanoma, will be an example of oncolytic vectors developed against rare cancers. Yescarta and Kymriah will showcase the use of retrovirus and lentivirus vectors in the autologous ex vivo production of chimeric antigen receptor T cells (CAR-T), licenced for the treatment of refractory leukaemias and lymphomas. Similar retroviral and lentiviral technology can be applied to autologous haematopoietic stem cells, exemplified by Strimvelis and Zynteglo, licenced treatments for adenosine deaminase-severe combined immunodeficiency (ADA-SCID) and β-thalassaemia respectively. Antisense oligonucleotide technologies will be highlighted through Onpattro and Tegsedi, RNA interference drugs licenced for familial transthyretin (TTR) amyloidosis, and Spinraza, a splice-switching treatment for spinal muscular atrophy (SMA). An initial comparison of the effectiveness of AAV and oligonucleotide therapies in SMA is possible with Zolgensma, an AAV serotype 9 vector, and Spinraza. Through these examples of marketed gene therapies and gene cell therapies, we will discuss the expanding applications of such novel technologies to previously intractable rare diseases

Sticky central limit theorems on open books

Given a probability distribution on an open book (a metric space obtained by gluing a disjoint union of copies of a half-space along their boundary hyperplanes), we define a precise concept of when the Fr\'{e}chet mean (barycenter) is sticky. This nonclassical phenomenon is quantified by a law of large numbers (LLN) stating that the empirical mean eventually almost surely lies on the (codimension $1$ and hence measure $0$) spine that is the glued hyperplane, and a central limit theorem (CLT) stating that the limiting distribution is Gaussian and supported on the spine. We also state versions of the LLN and CLT for the cases where the mean is nonsticky (i.e., not lying on the spine) and partly sticky (i.e., is, on the spine but not sticky).Comment: Published in at http://dx.doi.org/10.1214/12-AAP899 the Annals of Applied Probability (http://www.imstat.org/aap/) by the Institute of Mathematical Statistics (http://www.imstat.org

A Categorical Equivalence between Generalized Holonomy Maps on a Connected Manifold and Principal Connections on Bundles over that Manifold

A classic result in the foundations of Yang-Mills theory, due to J. W. Barrett ["Holonomy and Path Structures in General Relativity and Yang-Mills Theory." Int. J. Th. Phys. 30(9), (1991)], establishes that given a "generalized" holonomy map from the space of piece-wise smooth, closed curves based at some point of a manifold to a Lie group, there exists a principal bundle with that group as structure group and a principal connection on that bundle such that the holonomy map corresponds to the holonomies of that connection. Barrett also provided one sense in which this "recovery theorem" yields a unique bundle, up to isomorphism. Here we show that something stronger is true: with an appropriate definition of isomorphism between generalized holonomy maps, there is an equivalence of categories between the category whose objects are generalized holonomy maps on a smooth, connected manifold and whose arrows are holonomy isomorphisms, and the category whose objects are principal connections on principal bundles over a smooth, connected manifold. This result clarifies, and somewhat improves upon, the sense of "unique recovery" in Barrett's theorems; it also makes precise a sense in which there is no loss of structure involved in moving from a principal bundle formulation of Yang-Mills theory to a holonomy, or "loop", formulation.Comment: 20 page

The Pour-through Procedure Preferentially Extracts Substrate Solution from the Bottom of the Container in Conventional and Stratified Substrates

Due to the widespread use of the pour-through extraction procedure in horticultural production and research, the objective of this study was to determine if the method is biased by preferentially extracting substrate solution near the bottom of the container in both conventionally filled containers as well as intentionally stratified containers. Eight treatments were created using 2.5-L, 17.5-cm tall plastic nursery containers. The first four treatments were created by layering a conventional pine bark substrate (CONV) that was either amended (+A) or nonamended (−A) with fertilizer and lime with the following layers: amended substrate throughout the entire container profile (+A/+A); amended substrate in the top half (top 8.5 cm) over nonamended substrate in the bottom half of the container profile (+A/−A); nonamended substrate in the top half over amended substrate in the bottom half (−A/+A); and nonamended substrate throughout the profile (−A/−A). An additional four treatments were created by intentionally stratifying (STRAT) a fine pine bark substrate (FINE) over a coarse pine bark substrate (CRSE) with the same amendment combinations of +A or −A. On 0 and 42 d after potting, substrate pH and electrical conductivity (EC) were determined on samples collected by the pour-through procedure and 1:1 water extracts of the top and bottom layers in the container. At 42 d after potting, nutrient ions (NO3−, PO42−, K, Ca, Mg, and SO42−) were also measured in both pour-through and 1:1 water extracts of the top and bottom layers. At both dates and in both CONV and STRAT containers, pour-through substrate pH and EC more closely reflected those measurements in the bottom half of the container as determined by the 1:1 water extract. At 42 d after potting, nutrient ions determined by the pour-through procedure were more highly correlated to the 1:1 water extracts from the bottom half of the container compared with the top half of the container in both CONV and STRAT substrates. Evidence herein demonstrates that the pour-through procedure is more reflective of the lower half of the container than the upper half for both CONV and stratified substrates

The strength of sintered and adhesively bonded zirconia/veneer-ceramic bilayers

AbstractObjectivesRecently all-ceramic restorative systems have been introduced that use CAD/CAM technology to fabricate both the Y-TZP core and veneer-ceramic layers. The aim was to identify whether the CAD/CAM approach resulted in more favourable stressing patterns in the veneer-ceramic when compared with a conventionally sintered Y-TZP core/veneer-ceramic.MethodsNominally identical Vita VM9 veneer-ceramic disc-shaped specimens (0.7mm thickness, 12mm diameter) were fabricated. 20 specimens received a surface coating of resin-cement (Panavia 21); 20 specimens were bonded with the resin-cement to fully sintered Y-TZP (YZ Vita Inceram Vita) discs (0.27mm thickness, 12mm diameter). A final series of 20 Y-TZP core/veneer-ceramic specimens were manufactured using a conventional sintering route. Biaxial flexure strength was determined in a ball-on-ring configuration and stress at the fracture origin calculated using multilayer closed-form analytical solutions. Fractography was undertaken using scanning electron microscopy. The experimental test was simulated using Finite Element Analysis. Group mean BFS were compared using a one-way ANOVA and post hoc Tukey tests at a 95% significance level.ResultsResin cement application resulted in significant strengthening of the veneer-ceramic and further significant strengthening of the veneer-ceramic (p<0.01) occurred following bonding to the Y-TZP core. The BFS calculated at the failure origin for conventionally sintered specimens was significantly reduced when compared with the adhesively bonded Y-TZP/veneer-ceramic.ConclusionsUnder the test conditions employed adhesive cementation between CAD/CAM produced Y-TZP/veneer-ceramic layers appears to offer the potential to induce more favourable stress states within the veneer-ceramic when compared with conventional sintered manufacturing routes.Clinical significanceThe current investigation suggests that the stressing patterns that arise in all-ceramic restorations fabricated using CAD/CAM for both the core and veneer-ceramic layers differ from those that occur in conventionally sintered bilayer restorations. Further work is required to ascertain whether such differences will translate into improved clinical outcomes

Molecular gut content analysis of different spider body parts

Molecular gut-content analysis has revolutionized the study of food webs and feeding interactions, allowing the detection of prey DNA within the gut of many organisms. However, successful prey detection is a challenging procedure in which many factors affect every step, starting from the DNA extraction process. Spiders are liquid feeders with branched gut diverticula extending into their legs and throughout the prosoma, thus digestion takes places in different parts of the body and simple gut dissection is not possible. In this study, we investigated differences in prey detectability in DNA extracts from different parts of the spider's body: legs, prosoma and opisthosoma, using prey-specific PCR and metabarcoding approaches. We performed feeding trials with the woodlouse hunter spider Dysdera verneaui Simon, 1883 (Dysderidae) to estimate the time at which prey DNA is detectable within the predator after feeding. Although we found that all parts of the spider body are suitable for gut-content analysis when using prey-specific PCR approach, results based on metabarcoding suggested the opisthosoma is optimal for detection of predation in spiders because it contained the highest concentration of prey DNA for longer post feeding periods. Other spiders may show different results compared to D. verneaui, but given similarities in the physiology and digestion in different families, it is reasonable to assume this to be common across species and this approach having broad utility across spiders.Peer reviewe

Generalized r-Modes of the Maclaurin Spheroids

Analytical solutions are presented for a class of generalized r-modes of rigidly rotating uniform density stars---the Maclaurin spheroids---with arbitrary values of the angular velocity. Our analysis is based on the work of Bryan; however, we derive the solutions using slightly different coordinates that give purely real representations of the r-modes. The class of generalized r-modes is much larger than the previously studied `classical' r-modes. In particular, for each l and m we find l-m (or l-1 for the m=0 case) distinct r-modes. Many of these previously unstudied r-modes (about 30% of those examined) are subject to a secular instability driven by gravitational radiation. The eigenfunctions of the `classical' r-modes, the l=m+1 case here, are found to have particularly simple analytical representations. These r-modes provide an interesting mathematical example of solutions to a hyperbolic eigenvalue problem.Comment: 12 pages, 3 figures; minor changes and additions as will appear in the version to be published in Physical Review D, January 199