Role of PTHrP in human intestinal Caco-2 cell response to oxidative stress

AbstractWe have previously demonstrated that parathyroid hormone (PTH) induces apoptosis in human colon adenocarcinoma Caco-2 cells but the effects of its tumoral analog PTH-related peptide (PTHrP) in this cell line are still unknown. In the present work we investigated whether PTHrP, as PTH, is able to induce Caco-2 cell apoptosis or if it exerts protective effects under apoptotic conditions. Using Caco-2 cells cultured under serum deprivation in the presence or absence of PTHrP we demonstrated that, differently to PTH, its analog employed at the same concentration (10−8M) is not a pro-apoptotic hormone. Cells were exposed to an oxidative insult in the form of hydrogen peroxide to induce apoptosis, which leads to a 50% loss of cell viability determined by MTS assay, morphological changes observed under fluorescence microscopy and Western blot analysis. Herein we demonstrate, for the first time, that pre-treatment with PTHrP prior to H2O2 incubation, prevents cell death induced by the apoptotic inductor; and using specific inhibitors we evidenced that protein kinase B (AKT), extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase 1/2 (JNK1/2) and p38 mitogen-activated protein kinase (MAPK) mediate this anti-apoptotic effect. Also, we found that PTHrP decreases the pro-apoptotic protein BAX levels and increases the protein expression of the anti-apoptotic HSP27. Immunoblot analysis revealed that H2O2 increases the phosphorylation levels of AKT and MAPKs, exhibiting a cellular defense response; and consequently increases phospho-BAD levels. The H2O2-induced activation of protein kinases is reverted when cells are pre-treated with PTHrP. Altogether these results evidence a protective effect of PTHrP under apoptotic conditions in intestinal cells, which may be mediated by AKT and MAPKs

Impact of Natural Blind Spot Location on Perimetry.

We study the spatial distribution of natural blind spot location (NBSL) and its impact on perimetry. Pattern deviation (PD) values of 11,449 reliable visual fields (VFs) that are defined as clinically unaffected based on summary indices were extracted from 11,449 glaucoma patients. We modeled NBSL distribution using a two-dimensional non-linear regression approach and correlated NBSL with spherical equivalent (SE). Additionally, we compared PD values of groups with longer and shorter distances than median, and larger and smaller angles than median between NBSL and fixation. Mean and standard deviation of horizontal and vertical NBSL were 14.33° ± 1.37° and -2.06° ± 1.27°, respectively. SE decreased with increasing NBSL (correlation: r = -0.14, p \u3c 0.001). For NBSL distances longer than median distance (14.32°), average PD values decreased in the upper central (average difference for significant points (ADSP): -0.18 dB) and increased in the lower nasal VF region (ADSP: 0.14 dB). For angles in the direction of upper hemifield relative to the median angle (-8.13°), PD values decreased in lower nasal (ADSP: -0.11 dB) and increased in upper temporal VF areas (ADSP: 0.19 dB). In conclusion, we demonstrate that NBSL has a systematic effect on the spatial distribution of VF sensitivity

Design and Validation of an Open-Hardware Print-Head for Bioprinting Application

In the last decades drop-on-demand inkjet technology played an increasing role in industrial and medical applications. This is due to the ability to deposit a small amount of material in precisely defined position. In the field of Biofabrication, inkjet printers are used to build 2D and 3D scaffolds and gels with biological molecules, including living cells. Several works, including seminal papers on inkjet bioprinting, were carried out with modified office printers. These printers have fixed structural characteristics and operating size, especially on the print-head, limiting the range of materials that can be dispensed. The aim of the present work is the design and fabrication of an open-source piezoelectric inkjet print-head, optimized for the bioprinting field. This low-cost, reproducible, reliable, versatile and biocompatible device will enable various research laboratories to work with a shared device; the open source allowing for parts to be modified to suit specific needs. The design was carried out by Finite Element (FE) modelling of the piezoelectric, mechanical, fluid dynamics and their coupling. The design was optimized for shear rate, which we minimized in order to be able to print cells. The mechanical frame of the printer was designed and built using a low-cost 3D printer. The nozzle plate was fabricated from a polycarbonate disc coated with biocompatible silicone, to increase the hydrophobicity of the outer surface of the disc, preventing ink adhesion on the edge of the nozzle; the refilling system, and the electronic control were also part of the project and will be freely available to download. The FE models were validated with ad-hoc experiments, printing water, gelatin solution, and cell culture media, by modulating the wave power in amplitude, frequency and duty cycle. The tests showed a large working window both respect to viscosity and to surface tension. Finally Human Skin Fibroblasts (ATCC-CRL- 2522, Teddington UK), suspended in culture media, were printed. Cell viability, assessed by CellTiter-Blue and LIVE / DEAD tests, resulted comparable with the control, demonstrating the validity of the first open source piezoelectric inkjet print-head for biofabrication

An Artificial Intelligence Approach to Detect Visual Field Progression in Glaucoma Based on Spatial Pattern Analysis.

Purpose: To detect visual field (VF) progression by analyzing spatial pattern changes. Methods: We selected 12,217 eyes from 7360 patients with at least five reliable 24-2 VFs and 5 years of follow-up with an interval of at least 6 months. VFs were decomposed into 16 archetype patterns previously derived by artificial intelligence techniques. Linear regressions were applied to the 16 archetype weights of VF series over time. We defined progression as the decrease rate of the normal archetype or any increase rate of the 15 VF defect archetypes to be outside normal limits. The archetype method was compared with mean deviation (MD) slope, Advanced Glaucoma Intervention Study (AGIS) scoring, Collaborative Initial Glaucoma Treatment Study (CIGTS) scoring, and the permutation of pointwise linear regression (PoPLR), and was validated by a subset of VFs assessed by three glaucoma specialists. Results: In the method development cohort of 11,817 eyes, the archetype method agreed more with MD slope (kappa: 0.37) and PoPLR (0.33) than AGIS (0.12) and CIGTS (0.22). The most frequently progressed patterns included decreased normal pattern (63.7%), and increased nasal steps (16.4%), altitudinal loss (15.9%), superior-peripheral defect (12.1%), paracentral/central defects (10.5%), and near total loss (10.4%). In the clinical validation cohort of 397 eyes with 27.5% of confirmed progression, the agreement (kappa) and accuracy (mean of hit rate and correct rejection rate) of the archetype method (0.51 and 0.77) significantly (P \u3c 0.001 for all) outperformed AGIS (0.06 and 0.52), CIGTS (0.24 and 0.59), MD slope (0.21 and 0.59), and PoPLR (0.26 and 0.60). Conclusions: The archetype method can inform clinicians of VF progression patterns

Levy Anomalous Diffusion and Fractional Fokker--Planck Equation

We demonstrate that the Fokker-Planck equation can be generalized into a 'Fractional Fokker-Planck' equation, i.e. an equation which includes fractional space differentiations, in order to encompass the wide class of anomalous diffusions due to a Levy stable stochastic forcing. A precise determination of this equation is obtained by substituting a Levy stable source to the classical gaussian one in the Langevin equation. This yields not only the anomalous diffusion coefficient, but a non trivial fractional operator which corresponds to the possible asymmetry of the Levy stable source. Both of them cannot be obtained by scaling arguments. The (mono-) scaling behaviors of the Fractional Fokker-Planck equation and of its solutions are analysed and a generalization of the Einstein relation for the anomalous diffusion coefficient is obtained. This generalization yields a straightforward physical interpretation of the parameters of Levy stable distributions. Furthermore, with the help of important examples, we show the applicability of the Fractional Fokker-Planck equation in physics.Comment: 22 pages; To Appear in Physica

The liminality of trajectory shifts in institutional entrepreneurship

In this paper, we develop a process model of trajectory shifts in institutional entrepreneurship. We focus on the liminal periods experienced by institutional entrepreneurs when they, unlike the rest of the organization, recognize limits in the present and seek to shift a familiar past into an unfamiliar and uncertain future. Such periods involve a situation where the new possible future, not yet fully formed, exists side-by-side with established innovation trajectories. Trajectory shifts are moments of truth for institutional entrepreneurs, but little is known about the underlying mechanisms of how entrepreneurs reflectively deal with liminality to conceive and bring forth new innovation trajectories. Our in-depth case study research at CarCorp traces three such mechanisms (reflective dissension, imaginative projection, and eliminatory exploration) and builds the basis for understanding the liminality of trajectory shifts. The paper offers theoretical implications for the institutional entrepreneurship literature

Persistence, extinction and spatio-temporal synchronization of SIRS cellular automata models

Spatially explicit models have been widely used in today's mathematical ecology and epidemiology to study persistence and extinction of populations as well as their spatial patterns. Here we extend the earlier work--static dispersal between neighbouring individuals to mobility of individuals as well as multi-patches environment. As is commonly found, the basic reproductive ratio is maximized for the evolutionary stable strategy (ESS) on diseases' persistence in mean-field theory. This has important implications, as it implies that for a wide range of parameters that infection rate will tend maximum. This is opposite with present results obtained in spatial explicit models that infection rate is limited by upper bound. We observe the emergence of trade-offs of extinction and persistence on the parameters of the infection period and infection rate and show the extinction time having a linear relationship with respect to system size. We further find that the higher mobility can pronouncedly promote the persistence of spread of epidemics, i.e., the phase transition occurs from extinction domain to persistence domain, and the spirals' wavelength increases as the mobility increasing and ultimately, it will saturate at a certain value. Furthermore, for multi-patches case, we find that the lower coupling strength leads to anti-phase oscillation of infected fraction, while higher coupling strength corresponds to in-phase oscillation.Comment: 12page

At-line boar taint classification by means of Rapid Evaporative Ionisation Mass Spectrometry (REIMS)

Increasing awareness of animal welfare has led to a European incentive to ban the surgical castration of piglets. A valid alternative for castration is the rearing of entire male pigs, but this allows the (re)occurrence of boar taint, an off-odour in meat from entire boars (1). Hence, due to adverse consumer reactions to pork with boar taint, the rearing of entire boars requires valid boar taint mitigation strategies. However, the introduction of Rapid Evaporative Ionisation MS (REIMS) offers compelling perspectives for the rapid as well as accurate at-line detection of boar taint by significantly reducing analysis time and workload, yet enhancing research output and efficiency (2). In this study, REIMS was used as a direct analysis technique to train predictive models for identification of boar taint above the odour threshold (based on sensory (soldering iron method) as well as chemical analysis (UHPLC-HRMS analysis of indole, skatole and androstenone levels) 3. Adipose tissue was sampled using a prototype bipolar handheld sampling device connected directly to a Xevo G2-XS Q-TOF system equipped with REIMS source (Fig. 1). The results demonstrate that untargeted mass spectrometric profiling in negative ionisation mode enables the construction of predictive models using LiveID, AMX and Simca (Q2 = 0.547, R2Y = 0.652 and p = 0) for the classification of carcasses according to boar taint status based on alterations in lipid profiles. As REIMS eliminates sample pre-treatment with analysis taking < 10 seconds, it offers significant potential as the first technique enabling accurate in-situ detection of boar taint. REIMS is a promising and highly innovative tool for several types of food quality and safety applications, furthermore allowing us to move state-of-the-art equipment and applications from bench to production site. Acknowledgments: This research was partly funded by the Flemish Government, department of animal welfare (LNE/STG/DWZ/16/11). References: [1] K. Verplanken, Ghent University PhD dissertation, (2018) 1-15. [2] K. Verplanken et al., Talanta, 169 (2017) 30–36. [3] K. Bekaert et al., Journal of Chromatography A, 1239 (2012) 49-55