Calculating Colimits Compositionally

We show how finite limits and colimits can be calculated compositionally using the algebras of spans and cospans, and give as an application a proof of the Kleene Theorem on regular languages

Identification of Ilarviruses in almond and cherry fruit trees using nested PCR assays

In this study nested PCR assays have been developed for the detection of Prune dwarf virus (PDV), Prunus necrotic ringspot virus (PNRSV) and Apple mosaic virus (ApMV) modifying a previously reported assay for the generic detection of ilarviruses. In all cases one generic upstream primer was used along with a virus-specific downstream primer in respective nested PCR assays. The application of the same thermocycling profile allowed all amplifications to run in parallel. Ilarvirus isolates from different hosts were used for the evaluation of the detection range of the assays, which were afterwards applied for screening almond and cherry plant material. In almond trees the incidence of PNRSV and PDV was 41% and 21.5%, respectively. In cherry orchards the opposite was observed with PDV (56.6%) being the prevalent virus followed by PNRSV (19.4%). Mixed infections with both viruses were also encountered in approximately 10 and 17% of cherry and almond trees, respectively. ApMV was not detected in any of the samples tested. This is the first extensive survey conducted in Greece in order to monitor the distribution of these viruses using molecular assays. Keywords: Prune dwarf virus, Prunus necrotic ringspot virus, Apple mosaic virus, cherry, almond, nested PC

FAS2FURIOUS: Moderate-Throughput Secreted Expression of Difficult Recombinant Proteins in <em>Drosophila</em> S2 Cells

Copyright \ua9 2022 Coker, Katis, Fairhead, Schwenzer, Clemmensen, Frandsen, de Jongh, Gileadi, Burgess-Brown, Marsden, Midwood and Yue. Recombinant protein expression in eukaryotic insect cells is a powerful approach for producing challenging targets. However, due to incompatibility with standard baculoviral platforms and existing low-throughput methodology, the use of the Drosophila melanogaster “S2” cell line lags behind more common insect cell lines such as Sf9 or High-Five™. Due to the advantages of S2 cells, particularly for secreted and secretable proteins, the lack of a simple and parallelizable S2-based platform represents a bottleneck, particularly for biochemical and biophysical laboratories. Therefore, we developed FAS2FURIOUS, a simple and rapid S2 expression pipeline built upon an existing low-throughput commercial platform. FAS2FURIOUS is comparable in effort to simple E. coli systems and allows users to clone and test up to 46 constructs in just 2 weeks. Given the ability of S2 cells to express challenging targets, including receptor ectodomains, secreted glycoproteins, and viral antigens, FAS2FURIOUS represents an attractive orthogonal approach for protein expression in eukaryotic cells

Regulation of inositol 5-phosphatase activity by the C2 domain of SHIP1 and SHIP2

SHIP1, an inositol 5-phosphatase, plays a central role in cellular signaling. As such, it has been implicated in many conditions. Exploiting SHIP1 as a drug target will require structural knowledge and the design of selective small molecules. We have determined apo, and magnesium and phosphate-bound structures of the phosphatase and C2 domains of SHIP1. The C2 domains of SHIP1 and the related SHIP2 modulate the activity of the phosphatase domain. To understand the mechanism, we performed activity assays, hydrogen-deuterium exchange mass spectrometry, and molecular dynamics on SHIP1 and SHIP2. Our findings demonstrate that the influence of the C2 domain is more pronounced for SHIP2 than SHIP1. We determined 91 structures of SHIP1 with fragments bound, with some near the interface between the two domains. We performed a mass spectrometry screen and determined four structures with covalent fragments. These structures could act as starting points for the development of potent, selective probes

Varying numbers of players in small-sided soccer games modifies action opportunities during training

This study examined the effects of the numbers of players involved in small-sided team games (underloading and overloading) on opportunities for maintaining ball possession, shooting at goal and passing to teammates during training. These practice constraint manipulations were assumed to alter values of key performance variables identified in previous research, such as interpersonal distances between players and time to intercept shots and passes. Fifteen male soccer players (age: 19.60±1.99 years) were grouped into three teams and played against each other in different versions of small-sided soccer games, in which the number of players was manipulated in three different conditions: 5 vs. 5, 5 vs. 4 and 5 vs. 3. Dependent variables were the values of interpersonal distance between an outfield attacker and nearest defender (ID), and the relative distance of a defender needed to intercept the trajectory of a shot (RDishot) or pass (RDipass). Statistical analyses revealed that mean ID values were significantly lower in 5 vs. 5 than in 5 vs. 4 and 5 vs. 3 conditions, and significantly lower in 5 vs. 4 than 5 vs. 3. They also revealed that mean values of RDishot were significantly higher in 5 vs. 3 than in 5 vs. 5 conditions. Finally, results showed that the mean values of RDipass were significantly higher in 5 vs. 3 than in 5 vs. 5. Findings revealed how task constraints in SSGs can be manipulated to vary values of key spatial and temporal performance variables (interpersonal distance and time to intercept) to influence the nature of interpersonal interactions between competing players during practice. We observed that these manipulations tended to decrease opportunities for maintaining ball possession during training when equal numbers of attackers and defenders existed in SSGs, and led to more shots and passes emerging when the number of defenders was decreased relative to attackers

Parameterized Synthesis with Safety Properties

Parameterized synthesis offers a solution to the problem of constructing correct and verified controllers for parameterized systems. Such systems occur naturally in practice (e.g., in the form of distributed protocols where the amount of processes is often unknown at design time and the protocol must work regardless of the number of processes). In this paper, we present a novel learning based approach to the synthesis of reactive controllers for parameterized systems from safety specifications. We use the framework of regular model checking to model the synthesis problem as an infinite-duration two-player game and show how one can utilize Angluin's well-known L* algorithm to learn correct-by-design controllers. This approach results in a synthesis procedure that is conceptually simpler than existing synthesis methods with a completeness guarantee, whenever a winning strategy can be expressed by a regular set. We have implemented our algorithm in a tool called L*-PSynth and have demonstrated its performance on a range of benchmarks, including robotic motion planning and distributed protocols. Despite the simplicity of L*-PSynth it competes well against (and in many cases even outperforms) the state-of-the-art tools for synthesizing parameterized systems.Comment: 18 page

Discovery of FERM domain protein-protein interaction inhibitors for MSN and CD44 as a potential therapeutic approach for Alzheimer\u27s disease.

Proteomic studies have identified moesin (MSN), a protein containing a four-point-one, ezrin, radixin, moesin (FERM) domain, and the receptor CD44 as hub proteins found within a coexpression module strongly linked to Alzheimer\u27s disease (AD) traits and microglia. These proteins are more abundant in Alzheimer\u27s patient brains, and their levels are positively correlated with cognitive decline, amyloid plaque deposition, and neurofibrillary tangle burden. The MSN FERM domain interacts with the phospholipid phosphatidylinositol 4,5-bisphosphate (PI

Novel Role of Phosphorylation-Dependent Interaction between FtsZ and FipA in Mycobacterial Cell Division

The bacterial divisome is a multiprotein complex. Specific protein-protein interactions specify whether cell division occurs optimally, or whether division is arrested. Little is known about these protein-protein interactions and their regulation in mycobacteria. We have investigated the interrelationship between the products of the Mycobacterium tuberculosis gene cluster Rv0014c-Rv0019c, namely PknA (encoded by Rv0014c) and FtsZ-interacting protein A, FipA (encoded by Rv0019c) and the products of the division cell wall (dcw) cluster, namely FtsZ and FtsQ. M. smegmatis strains depleted in components of the two gene clusters have been complemented with orthologs of the respective genes of M. tuberculosis. Here we identify FipA as an interacting partner of FtsZ and FtsQ and establish that PknA-dependent phosphorylation of FipA on T77 and FtsZ on T343 is required for cell division under oxidative stress. A fipA knockout strain of M. smegmatis is less capable of withstanding oxidative stress than the wild type and showed elongation of cells due to a defect in septum formation. Localization of FtsQ, FtsZ and FipA at mid-cell was also compromised. Growth and survival defects under oxidative stress could be functionally complemented by fipA of M. tuberculosis but not its T77A mutant. Merodiploid strains of M. smegmatis expressing the FtsZ(T343A) showed inhibition of FtsZ-FipA interaction and Z ring formation under oxidative stress. Knockdown of FipA led to elongation of M. tuberculosis cells grown in macrophages and reduced intramacrophage growth. These data reveal a novel role of phosphorylation-dependent protein-protein interactions involving FipA, in the sustenance of mycobacterial cell division under oxidative stress