Herein, we report the use of a versatile photocleavable nitrobenzyl linker to micropattern a wide variety of bioactive molecules and photorelease them on demand. On one end, the linker has an NHS ...group that can be coupled with any amine, such as peptides, proteins or amine-linkers, and on the other end an alkyne for convenient attachment to materials with an azide functional group. This linker was conjugated with NTA-amine or the cell adhesion peptide cRGD to enable straightforward patterning of His6-tagged proteins or cells, respectively, on PEGylated glass surfaces. This approach provides a practical way to control the presentation of a wide variety of bioactive molecules with high spatial and temporal resolution. The extent of photocleavage can also be controlled to tune the biomolecule density and degree of cell attachment to the surface.
Developing orthogonal chemical communication pathways in diverse synthetic cell communities is a considerable challenge due to the increased crosstalk and interference associated with large numbers ...of different types of sender‐receiver pairs. Herein, the authors control which sender‐receiver pairs communicate in a three‐membered community of synthetic cells through red and blue light illumination. Semipermeable protein‐polymer‐based synthetic cells (proteinosomes) with complementary membrane‐attached protein adhesion communicate through single‐stranded DNA oligomers and synergistically process biochemical information within a community consisting of one sender and two different receiver populations. Different pairs of red and blue light‐responsive protein‐protein interactions act as membrane adhesion mediators between the sender and receivers such that they self‐assemble and socially self‐sort into different multicellular structures under red and blue light. Consequently, distinct sender‐receiver pairs come into the signaling range depending on the light illumination and are able to communicate specifically without activation of the other receiver population. Overall, this work shows how photoswitchable membrane adhesion gives rise to different self‐sorting protocell patterns that mediate member‐specific DNA‐based communication in ternary populations of synthetic cells and provides a step towards the design of orthogonal chemical communication networks in diverse communities of synthetic cells.
Herein, the self‐assembly and social self‐sorting into different multicellular in a three‐membered community of synthetic cells under red and blue light are controlled. Consequently, distinct sender‐receiver pairs come into the signaling range depending on the light illumination and are able to communicate specifically without activation of the other receiver population.
The Fermi Gamma-ray Space Telescope (Fermi) is producing the most detailed inventory of the gamma-ray sky to date. Despite tremendous achievements approximately 25 per cent of all Fermi extragalactic ...sources in the Second Fermi Large Area Telescope Catalogue (2FGL) are listed as active galactic nuclei (AGN) of uncertain type. Typically, these are suspected blazar candidates without a conclusive optical spectrum or lacking spectroscopic observations. Here, we explore the use of machine-learning algorithms - random forests and support vector machines - to predict specific AGN subclass based on observed gamma-ray spectral properties. After training and testing on identified/associated AGN from the 2FGL we find that 235 out of 269 AGN of uncertain type have properties compatible with gamma-ray BL Lacertae and flat-spectrum radio quasars with accuracy rates of 85 per cent. Additionally, direct comparison of our results with class predictions made after following the infrared colour-colour space of Massaro et al. shows that the agreement rate is over four-fifths for 54 overlapping sources, providing independent cross-validation. These results can help tailor follow-up spectroscopic programmes and inform future pointed surveys with ground-based Cherenkov telescopes.
In this paper, we introduce two methods for determining feeding buffer sizes in critical chain project scheduling. Both methods integrate project characteristics into the formulation. Specifically, ...one of them incorporates resource tightness while the other uses network complexity. Both methods are tested and compared to two commonly suggested methods in the literature, the cut and paste method and the root square error method, as well as using no buffer as a benchmark. The comparison is done by means of a simulation study using the Patterson data set. The test results indicate that both of the suggested methods generate smaller buffer sizes while providing sufficient protection against delays in project completion time.
We present a detailed analysis of the interstellar medium towards the tera electron volt (TeV) ...-ray sources HESS J1640-465 and HESS J1641-463 using results from the Mopra Southern Galactic Plane ...CO Survey and from a Mopra 7 mm-wavelength study. The ...-ray sources are positionally coincident with two supernova remnants (SNRs) G338.3-0.0 and G338.5+0.1, respectively. A bright complex of H ii regions connect the two SNRs and TeV objects. Observations in the CO(1-0) transition lines reveal substantial amounts of diffuse gas positionally coincident with the ...-ray sources at multiple velocities along the line of sight, while 7 mm observations in CS, SiO, HC sub( 3)N and CH sub( 3)OH transition lines reveal regions of dense, shocked gas. Archival H i data from the Southern Galactic Plane Survey was used to account for the diffuse atomic gas. Physical parameters of the gas towards the TeV sources were calculated from the data. We find that for a hadronic origin for the ...-ray emission, the cosmic ray enhancement rates are ~10 super( 3) and 10 super( 2) times the local solar value for HESS J1640-465 and HESS J1641-463, respectively. (ProQuest: ... denotes formulae/symbols omitted.)
Introduction
We characterized five patients affected with von Willebrand disease (VWD) carrying the p.Arg1379Cys mutation. One was diagnosed as VWD type 1 and four as type 2M. The 2M patients also ...have the variant p.Ala1377Val in cis with p.Arg1379Cys.
Aim
To evaluate the role of p.Ala1377Val and p.Arg1379Cys von Willebrand factor (VWF) variants to explain patients' phenotype.
Methods
Conventional phenotype tests were used to evaluate patients' plasma and platelets. Direct sequence analysis of exon 28 was carried out. The allele frequency of p.Ala1377Val was evaluated using online database. pcDNA3.1‐VWF‐WT and mutant (A1377V, R1379C and A1377V‐R1379C) expression vectors were transiently transfected in HEK293 cells. The capacity of WT and mutant recombinant (r)VWF (along with patients' plasma VWF) to bind glycoprotein Ibα (GpIbα) were evaluated, using two ELISA assays. One with a wild‐type (WT) recombinant (r)GpIbα at increasing ristocetin concentrations (from 0 to 1.50 mg mL−1) and the other with a gain‐of‐function mutant rGpIbα (VWF:GPIbM).
Results
The substitution c.4130C>T (p.Ala1377Val) was reported as rare variant in online databases. At 0.25 mg mL−1 of ristocetin, WT, A1377V and R1379C showed 6, 7.5 and 12‐fold increased binding to rGpIbα, respectively. A1377V‐R1379C rVWF showed no increased binding to rGpIbα at the same ristocetin concentration and reached the highest binding, of only 3‐fold increased, at 1.50 mg mL−1 of ristocetin. The VWF:GPIbM showed strongly reduced values for the A1377V‐R1379C rVWF and the 2M patients' plasma.
Conclusion
Our study showed that the presence of both p.Ala1377Val and p.Arg1379Cys mutations (synergistic effect) abolishes the binding of rVWF to rGpIbα, explaining patients' 2M phenotype.
Summary
Background
Diagnosis of von Willebrand disease (VWD) type 2 usually relies on the discrepancy between the von Willebrand factor (VWF) ristocetin cofactor activity (VWF:RCo) and VWF antigen ...(VWF:Ag). Type 2B patients can be discriminated from other qualitative VWD variants by using ristocetin‐induced platelet agglutination (RIPA) test. The major limitation of RIPA is the requirement of fresh blood sample.
Objectives
In this study, we evaluated the VWF gain‐of‐function mutant GPIb binding (VWF:GPIbM) and VWF:RCo assays to investigate whether the VWF:GPIbM/VWF:RCo ratio was able to identify the type 2B variant among an heterogeneous VWD population, previously characterized following the ISTH‐SSC guidelines.
Patients/methods
Seventy‐six VWD patients and 31 healthy subjects were evaluated by using VWF:Ag, VWF:RCo, and VWF:GPIbM assays.
Results
The mean (minimum–maximum values) VWF:GPIbM/VWF:RCo ratio was higher in type 2B patients (2.53, 0.84–6.11) than in healthy controls (1.05, 0.87–1.34), type 1 (0.85, 0.51–1.15), 2A (1.20, 0.36–2.82), and 2M (1.07, 0.91–1.38) (P < 0.0001). Type 2B variants were divided into four groups (A, B, C, and D) according to their different multimeric patterns. The mean value of the VWF:GPIbM/VWF:RCo ratio in the four groups showed an increasing trend from group A (1.08) to D (3.69), proportional to the loss of high molecular weight multimers. Among 32 type 2B patients, previously diagnosed with RIPA, 8 (mainly with a type I New York/Malmö phenotype) were not confirmed using the VWF:GPIbM/VWF:RCo ratio.
Conclusions
Whenever the RIPA test is not feasible, the VWF:GPIbM/VWF:RCo ratio might help to identify severe type 2B VWD patients.
Promotion of neurite outgrowth is an important limiting step for regeneration in nerve injury and depends strongly on the local expression of nerve growth factor (NGF). The rational design of ...bioactive materials is a promising approach for the development of novel therapeutic methods for nerve regeneration, and biomaterials capable of presenting NGF to nerve cells are especially suitable for this purpose. In this study, we show bioactive peptide amphiphile (PA) nanofibers capable of promoting neurite outgrowth by displaying high density binding epitopes for NGF. A high-affinity NGF-binding sequence was identified by phage display and combined with a beta-sheet forming motif to produce a self-assembling PA molecule. The bioactive nanofiber had higher affinity for NGF compared to control nanofibers and in vitro studies revealed that the NGF binding peptide amphiphile nanofibers (NGFB-PA nanofiber) significantly promote the neurite outgrowth of PC-12 cells. In addition, the nanofibers induced differentiation of PC-12 cells into neuron-like cells by enhancing NGF/high-activity NGF receptor (TrkA) interactions and activating MAPK pathway elements. The NGFB-PA nanofiber was further shown as a promising material to support axonal outgrowth from primary sensory neurons. These materials will pave the way for the development of new therapeutic agents for peripheral nervous system injuries.
Purpose - The applicability of the customer life time value (CLV) concept goes beyond consumer markets. Specifically, the purpose of this paper is to show how a make-to-order manufacturing company in ...a supply chain can set customer-focus manufacturing strategies using CLV.Design methodology approach - Data from an integrated steel plant is used to calculate the life time value of customers based on the past value, the potential value, and their loyalty. The past value of a customer is based on the historical data and the future value of a customer is then forecasted. The loyalty index of a customer is determined by survey results.Findings - In general, it was found that the CLV for the most valuable customers increases exponentially and the top 28 percent of customers constitute 80 percent of the total value of all customers.Research limitations implications - This study focuses on make-to-order manufacturing organizations and the three strategies suggested for business process improvement need to be re-evaluated for make-to-stock or mass production.Practical implications - Based on these results, the authors suggest three strategies for business process improvement and revenue growth for the plant.Originality value - This study constitutes an initial effort to develop a CLV model for make-to-order manufacturing organizations for improving plant performance. The model links customers with not only the front office functions but also with ERP systems. Organizations that are part of value chains can benefit significantly from CLV applications.
Summary
Introduction
We characterized four unrelated patients with von Willebrand disease type 2A/IIE, sharing the same von Willebrand factor (VWF) in‐frame deletion (p.P1127_G1180delinsR;=) ...resulting from exon 26 skipping (Δ26).
Objectives
To identify the VWF mutations and how they caused the mRNA splicing alteration, to evaluate the deletion by in vitro expression studies, and to assess whether or not the heterogeneity of the patients’ phenotype might be related to a different degree of expression of the deleted subunit in patient plasma VWF.
Methods
Sequence analysis was performed with patient genomic DNA and platelet mRNA. Semiquantitative RT‐PCR was also carried out to compare the expression of the wild‐type (WT) and Δ26 alleles in the four patients. In silico analysis was performed with prediction splicing programs. Expression studies were performed to evaluate mutant recombinant VWF (rVWF) (Δ26 and Δ26/WT) as compared with WT rVWF.
Results
Three patients shared the synonymous single‐nucleotide substitution (SSS) c.3390C>T;=, whereas the novel mutation c.3380‐2A>G;= was present in the fourth patient. Semiquantitative RT‐PCR of platelet mRNA revealed a different ratio of the WT and Δ26 alleles in the patients, consistent with the different VWF:FVIIIB values present in patient plasma. Expression studies confirmed reduced VWF–FVIII binding of rVWF‐Δ26/WT.
Conclusions
SSS can induce alternative splicing, and those like c.3390C>T, which impact on the poorly conserved splicing regulatory elements, are difficult to predict, so that their role can be evaluated only by mRNA analysis. Moreover, these mutations seem to have different effects on the efficiency of alternative splicing, producing heterogeneous VWF variants among the four patients.
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