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With the objective of studying the effect of two nutrient solutions and two crop systems (greenhouse and openfield) on nitrate accumulation, incidence of tipburn and chlorophyll content, endive (cv. Cuartana) was planted in 8 L pots, filled with a mixture of coconut coir:perlite (1:1) in three different cycles C1 (winter), C2 (spring) and C3 (summer). Plants were irrigated with two nutrient solutions of different nitrate content: S1, low ([NO−3] = 7.91 mmol L−1) and S2 moderate nitrate content ([NO−3] = 16.91 mmol L−1). Nitrate content was determined by reflectometry, tipburn was evaluated using a qualitative scale and chlorophyll content by soil plant analysis development(SPAD) values. Plants irrigated with S2 showed higher nitrate accumulation in leaves in all cycles, however, no influence of the nutrient solution was observed on the incidence of tipburn. Greenhouse-cultivated plants accumulated more nitrates than those cultivated in open field and also showed a higher incidence of tipburn and SPAD values. [ABSTRACT FROM PUBLISHER]

Cape gooseberry (Physalis peruvianaL.) is a solanaceous plant. The growth and time-course of nutrient accumulation of the plant and its partitioning between roots, stems, leaves, and fruits were examined. The study was conducted analyzing two nutrient solutions in soilless culture under greenhouse conditions during two consecutive seasons. The macronutrient contents were analyzed. On average, the yield was 8.9 t·ha−1. Growth of the plant until 90 d after transplanting obeys an exponential function of time and the relative growth rate for this period was determined. Nitrogen (N) was the element that showed the highest concentration, corresponding to leaves (4.67%), followed by potassium (K) in stems (4.46%). The highest accumulations of N, phosphorous (P), calcium (Ca), and magnesium (Mg) were found in leaves and of K in the stems. Potassium showed the highest nutrient accumulation (29 g·plant−1) and the highest specific uptake rate. [ABSTRACT FROM PUBLISHER]

The plug plant technique for the commercial propagation of strawberries is increasing in importance. Several factors, including the properties of the substrate, can affect plug plant quality. Tests on nine substrates containing different proportions of perlite [from 0-75%, (v/v)], and dark and light peat [both from 0- 80% (v/v)], were performed using a simplex-lattice design in order to establish a model for strawberry plug plant production based not only on the single component composition of the substrate, but also on the influences of the chemical and physical properties of the substrate on plug plant quality. Notable differences in physical and chemical properties were found among the nine different substrates tested, as a consequence of the broad range of their component compositions. Substrate mixes containing medium-to-high proportions [from 60 - 70% (v/v)] of light peat and low proportions of dark peat and perlite are recommended, as these resulted in a low nutrient content, a high organic matter content, a low pH, and a low "coarseness" index, which led to high-grade plug plants with greater root and crown dry weights. [ABSTRACT FROM AUTHOR]

To determine their influence on leaf tipburn incidence and yields six nutrient solutions with different proportions of potassium (K), calcium (Ca), and magnesium (Mg) were tested in 'Camarosa' strawberry plants. Response models were made to predict tipburn incidence, yields and fruit weight. A high Ca proportion in the nutrient solution was determinant for high yields in the first season, while a high Mg content was fundamental for high marketable yields during the two last seasons. In 2001 and 2002, the average fruit weight was higher with high Mg levels, and in 2000 the lowest fruit weights coincided with low levels of Ca in the nutrient solution. The models for tipburn incidence in 2000 and 2002 indicated that solutions with intermediate to high levels of Mg and intermediate to low levels of Ca resulted in a more frequent incidence of tipburn. Conversely, in 2001, the highest tipburn incidence appeared with high K contents. [ABSTRACT FROM AUTHOR]

The efficiency of fertilisation in agriculture is often low and, provided one knows the nutrient uptake rate, the efficiency can be improved by synchronising nutrient supply with nutrient demand. Growth, the time-course of nutrient accumulation and its partitioning between different organs in chufa (Cyperus esculentus L. var. sativus Boeck.), an under-exploited cultivated plant, were examined. The study was conducted in soilless, open-field conditions, at a planting density equivalent to 55,500 plants ha1 in three consecutive seasons. Plants were sampled, fractionated into leaves, roots, and tubers, then dried and weighed. Their macronutrient contents were analysed each fortnight, On average, the yield was 5.0 kg fresh weight of tuber m-2. Growth of whole plants, until 90 d after planting, obeyed an exponential function of time. The relative growth rate (RGR) for this period was therefore determined. The highest N and K concentrations were recorded in leaves, and the highest P, Ca, and Mg concentrations were found in roots. The highest accumulations of N and P were found in tubers, and of K and Ca in leaves. Nitrogen had the highest nutrient accumulation (58.3 g m-2) as well as the highest specific uptake rate. [ABSTRACT FROM AUTHOR]

The effect on fruit quality of grafting and fruit set treatments, was determined in the triploid watermelon (Citrullus lanatus (Thun.) Matsum. and Nakai) cultivar 'Reina de Corazones' grown in a heated greenhouse. Grafting the scion onto 'Shintoza' (Cucurbita maxima × Cucurbita moschata) rootstock and setting fruit through application of the synthetic cytokinin CPPU [1-(2-chloro-4-pyridyl)-3-phenylurea] retarded the accumulation of sugar and development of the flesh colour of the fruit, compared to fruit from non-grafted, pollinated plants. The effects of grafting and of CPPU application on sugar accumulation were additive. At commercial maturity, the fruit of grafted and/or CPPU-treated plants had lower total soluble solids and sugar concentrations, and lower sucrose to hexose ratios, than fruit from control plants. Fruit from CPPU-treated plants also had a weaker red colour than fruit set by pollination. Despite this loss of quality, fruit from grafted plants, set with CPPU and grown in a heated greenhouse, had a total soluble solids concentration similar to fruit from pollinated, non-grafted plants, grown outdoors. [ABSTRACT FROM AUTHOR]

A two-year study was conducted to analyze the yield and irrigation water-use efficiency of chufa crop in response to planting configuration and drip irrigation scheduling as a function of the volumetric soil water content. The planting configurations were: beds with three plant rows and three driplines (B3), beds with three plant rows and two driplines (B2), beds with two plant rows and two driplines ( b ), and ridges ( R ). The yield was affected by the planting configuration; greater yields were obtained in beds (on average 2.36 kg m −2 ) than in R (2.14 kg m −2 ). Considerably less irrigation water was applied in R and in B2 than in beds B3 and b . The irrigation water-use efficiency was affected by the planting configuration in the same line that the irrigation water was applied, with greater values being obtained in B2 (7.58 kg m −3 ) than in the R (6.63 kg m −3 ), which in turn was higher than B3 (5.92 kg m −3 ) and b (5.69 kg m −3 ). These values of the irrigation water-use efficiency were considerably higher than those obtained in previous experiments (based on the volumetric soil water content in the ridges). Neither the yield nor the average tuber weight were affected by the position of the different planting rows in the bed. [ABSTRACT FROM AUTHOR]

High salinity decreases the productivity of crops worldwide. Pepper is particularly sensitive to high salt concentrations. Herein, we subjected three tolerant pepper accessions (C12, B14 and A25) to high sodium chloride concentration (70 mM NaCl). The aerial and root biomass, leaf and root osmotic potential (Ψ π ), Na + , Cl - , K + and proline concentrations and the relative expression of the putative genes CaSOS1, CaHKT1, three CaNHXs and CaP5CS were measured. Different salinity tolerance strategies depending on the pepper accession were identified. In C12, tolerance was attributed to the accumulation of Na + in vacuoles and endosomes by the activation of vacuolar CaNHXs genes and the reduction in Ψ π ; additionally, the activation of CaHKT1 and CaSOS1 in leaves and roots moved and accumulated Na + ions in the xylem and xylem parenchyma cells (XPC) as well as expulsed it out of the root cells. A25 accession, on the contrary, was specialized in compartmentalizing Na + ions in root and leaf vacuoles and root XPC by the up-regulation of CaNHXs and CaHKT1, respectively, avoiding a toxic accumulation in leaves. Finally, B14 accession moved and accumulated Na + in xylem and XPC, reducing its concentration in roots by the activation of CaSOS1 and CaHKT1. This study shade light on different tolerance mechanisms of pepper plants to overcome salt stress., (© 2023 Scandinavian Plant Physiology Society.)

In the current climate change scenario, water stress is a serious threat to limit crop growth and yields. It is necessary to develop tolerant plants that cope with water stress and, for this purpose, tolerance mechanisms should be studied. NIBER® is a proven water stress- and salt-tolerant pepper hybrid rootstock (Gisbert-Mullor et al., 2020; López-Serrano et al., 2020), but tolerance mechanisms remain unclear. In this experiment, NIBER® and A10 (a sensitive pepper accession (Penella et al., 2014)) response to short-term water stress at 5 h and 24 h was studied in terms of gene expression and metabolites content in roots. GO terms and gene expression analyses evidenced constitutive differences in the transcriptomic profile of NIBER® and A10, associated with detoxification systems of reactive oxygen species (ROS). Upon water stress, transcription factors like DREBs and MYC are upregulated and the levels of auxins, abscisic acid and jasmonic acid are increased in NIBER®. NIBER® tolerance mechanisms involve an increase in osmoprotectant sugars (i.e., trehalose, raffinose) and in antioxidants (spermidine), but lower contents of oxidized glutathione compared to A10, which indicates less oxidative damage. Moreover, the gene expression for aquaporins and chaperones is enhanced. These results show the main NIBER® strategies to overcome water stress., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Phytohormones play an important role in regulating the plant behavior to drought. In previous studies, NIBER ® pepper rootstock showed tolerance to drought in terms of production and fruit quality compared to ungrafted plants. In this study, our hypothesis was that short-term exposure to water stress in young, grafted pepper plants would shed light on tolerance to drought in terms of modulation of the hormonal balance. To validate this hypothesis, fresh weight, water use efficiency (WUE) and the main hormone classes were analyzed in self-grafted pepper plants (variety onto variety, V/V) and variety grafted onto NIBER ® (V/N) at 4, 24, and 48h after severe water stress was induced by PEG addition. After 48h, WUE in V/N was higher than in V/V, due to major stomata closure to maintain water retention in the leaves. This can be explained by the higher abscisic acid (ABA) levels observed in the leaves of V/N plants. Despite the interaction between ABA and the ethylene precursor, 1-aminocyclopropane-1-carboxylic acid (ACC), in relation to stomata closure is controversial, we observed an important increase of ACC at the end of the experiment in V/N plants coinciding with an important rise of the WUE and ABA. The maximum concentration of jasmonic acid and salicylic acid after 48h was found in the leaves of V/N, associated with their role in abiotic stress signaling and tolerance. Respect to auxins and cytokinins, the highest concentrations were linked to water stress and NIBER ® , but this effect did not occur for gibberellins. These results show that hormone balance was affected by water stress and rootstock genotype, where NIBER ® rootstock displayed a better ability to overcome short-term water stress., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Padilla, Gisbert-Mullor, López-Galarza, Albacete, Martínez-Melgarejo and Calatayud.)

Although parental socialization has an influence on child development, current research is questioning which combination of parental strictness and warmth acts as protective or risk factors, especially during adolescence when the child is more vulnerable. The sample was 2125 participants, 58.7% female, divided into four age groups: adolescents (28.57%), young adults (28.38%), middle-aged adults (23.95%), and older adults (19.11%). The families were classified into four parenting styles: neglectful, indulgent, authoritative, and authoritarian according to their warmth and strictness scores. The psychosocial adjustment was measured by children's scores on academic/professional self-concept, self-esteem, delinquency during adolescence, and benevolence values. A MANOVA 4 × 2 × 4 was applied with parenting styles, sex, and age group as independent variables. The results showed that, for adolescents and adult children, only parenting styles characterized by warmth (i.e., indulgent, and authoritative) were found to factor against delinquency during adolescence and benefit greater academic/professional self-concept, self-esteem, and benevolence values, while parenting without warmth (i.e., authoritarian, and neglectful) were identified as risk factors. Contrary to classical research, the present findings seriously question the universal benefits of strict parenting as the only optimal strategy to protect not only against delinquency, but also to foster an adequate self and the internalization of social values.

Quinoa is an important crop for food security and food sovereignty in Ecuador. In this study, we evaluated the nutritional value, bioactive compounds, and antinutrient compounds of leaves and grains of the Ecuadorian quinoa variety Tunkahuan, and we identified significant differences between the nutrient content in the leaves and grains. The quinoa leaves presented a higher protein content than the grains, as well as inorganic nutrients such as calcium, phosphorus, iron, and zinc. Both the grains and leaves had an appreciable phenolic content. In addition, the quinoa grains presented a higher content of the antinutrient saponin than the leaves, while the leaves contained more nitrates and oxalates than the grains. Thus, quinoa leaves and grains exhibit excellent potential for application in the food and pharmaceutical industries.

Currently, limited water supply is a major problem in many parts of the world. Grafting peppers onto adequate rootstocks is a sustainable technique used to cope with water scarcity in plants. For 1 month, this work compared grafted peppers by employing two rootstocks (H92 and H90), with different sensitivities to water stress, and ungrafted plants in biomass, photosynthesis, and antioxidant response terms to identify physiological-antioxidant pathways of water stress tolerance. Water stress significantly stunted growth in all the plant types, although tolerant grafted plants (variety grafted onto H92, Var/H92) had higher leaf area and fresh weight values. Var/H92 showed photosynthesis and stomata conductance maintenance, compared to sensitive grafted plants (Var/H90) and ungrafted plants under water stress, linked with greater instantaneous water use efficiency. The antioxidant system was effective in removing reactive oxygen species (ROS) that could damage photosynthesis; a significant positive and negative linear correlation was observed between the rate of CO 2 uptake and ascorbic acid (AsA)/total AsA (AsA t ) and proline, respectively. Moreover, in Var/H92 under water stress, both higher proline and ascorbate concentration were observed. Consequently, less membrane lipid peroxidation was quantified in Var/H92.

Background: Pepper is one of the most cultivated crops worldwide, but is sensitive to salinity. This sensitivity is dependent on varieties and our knowledge about how they can face such stress is limited, mainly according to a molecular point of view. This is the main reason why we decided to develop this transcriptomic analysis. Tolerant and sensitive accessions, respectively called A25 and A6, were grown for 14 days under control conditions and irrigated with 70 mM of NaCl. Biomass, different physiological parameters and differentially expressed genes were analysed to give response to differential salinity mechanisms between both accessions., Results: The genetic changes found between the accessions under both control and stress conditions could explain the physiological behaviour in A25 by the decrease of osmotic potential that could be due mainly to an increase in potassium and proline accumulation, improved growth (e.g. expansins), more efficient starch accumulation (e.g. BAM1), ion homeostasis (e.g. CBL9, HAI3, BASS1), photosynthetic protection (e.g. FIB1A, TIL, JAR1) and antioxidant activity (e.g. PSDS3, SnRK2.10). In addition, misregulation of ABA signalling (e.g. HAB1, ERD4, HAI3) and other stress signalling genes (e.g. JAR1) would appear crucial to explain the different sensitivity to NaCl in both accessions., Conclusions: After analysing the physiological behaviour and transcriptomic results, we have concluded that A25 accession utilizes different strategies to cope better salt stress, being ABA-signalling a pivotal point of regulation. However, other strategies, such as the decrease in osmotic potential to preserve water status in leaves seem to be important to explain the defence response to salinity in pepper A25 plants.

Changes in the elastic properties of brain tissue have been correlated with injury, cancers, and neurodegenerative diseases. However, discrepancies in the reported elastic moduli of brain tissue are persistent, and spatial inhomogeneities complicate the interpretation of macroscale measurements such as rheology. Here we introduce needle induced cavitation rheology (NICR) and volume-controlled cavity expansion (VCCE) as facile methods to measure the apparent Young's modulus E of minimally manipulated brain tissue, at specific tissue locations and with sub-millimeter spatial resolution. For different porcine brain regions and sections analyzed by NICR, we found E to be 3.7 ± 0.7 kPa and 4.8 ± 1.0 kPa for gray matter, and white matter, respectively. For different porcine brain regions and sections analyzed by VCCE, we found E was 0.76 ± 0.02 kPa for gray matter and 0.92 ± 0.01 kPa for white matter. Measurements from VCCE were more similar to those obtained from macroscale shear rheology (0.75 ± 0.06 kPa) and from instrumented microindentation of white matter (0.97 ± 0.40 kPa) and gray matter (0.86 ± 0.20 kPa). We attributed the higher stiffness reported from NICR to that method's assumption of a cavitation instability due to a neo-Hookean constitutive response, which does not capture the strain-stiffening behavior of brain tissue under large strains, and therefore did not provide appropriate measurements. We demonstrate via both analytical modeling of a spherical cavity and finite element modeling of a needle geometry, that this strain stiffening may prevent a cavitation instability. VCCE measurements take this stiffening behavior into account by employing an incompressible one-term Ogden model to find the nonlinear elastic properties of the tissue. Overall, VCCE afforded rapid and facile measurement of nonlinear mechanical properties of intact, healthy mammalian brain tissue, enabling quantitative comparison among brain tissue regions and also between species. Finally, accurate estimation of elastic properties for this strain stiffening tissue requires methods that include appropriate constitutive models of the brain tissue response, which here are represented by inclusion of the Ogden model in VCCE., (Copyright © 2020. Published by Elsevier Ltd.)

Linear and non-linear models used in applications of genomic selection (GS) can fit different types of responses (e.g., continuous, ordinal, binary). In recent years, several genomic-enabled prediction models have been developed for predicting complex traits in genomic-assisted animal and plant breeding. These models include linear, non-linear and non-parametric models, mostly for continuous responses and less frequently for categorical responses. Several linear and non-linear models are special cases of a more general family of statistical models known as artificial neural networks, which provide better prediction ability than other models. In this paper, we propose a Bayesian Regularized Neural Network (BRNNO) for modelling ordinal data. The proposed model was fitted using a Bayesian framework; we used the data augmentation algorithm to facilitate computations. The proposed model was fitted using the Gibbs Maximum a Posteriori and Generalized EM algorithm implemented by combining code written in C and R programming languages. The new model was tested with two real maize datasets evaluated for Septoria and GLS diseases and was compared with the Bayesian Ordered Probit Model (BOPM). Results indicated that the BRNNO model performed better in terms of genomic-based prediction than the BOPM model., (© 2020 The Authors. The Plant Genome published by Wiley Periodicals, LLC on behalf of Crop Science Society of America.)

Grafting is a widely utilized agronomical technique to improve yield, disease resistance, and quality of fruit and vegetables. This work aims to assess the effect of grafting and fruit ripening on the production, physico-chemical characteristics, and nutritional quality of fruit from Spanish local pepper landraces. Landraces "Cuerno," "Sueca," and "Valencia" were used as scions, and "NIBER ® " as the rootstock. Two ripening stages of the fruits were sampled: green and red. Grafting improved the yield and marketable quality and did not negatively influence the physico-chemical and nutritional characteristics of the fruit. It was noteworthy that the bioactive compound contents and antioxidant capacity were more related to maturity stage and genotype, and red fruit had a higher antioxidant capacity than green fruit. However, in all the scions, grafting significantly enhanced lycopene content in both red and green fruit. Another important effect of grafting was the volatile compound composition evidenced by discriminant analyses, which was characterized for the first time in the fruit of these landraces. The rootstock and scion combination could be a way to improve not only the production, but also the fruit quality of peppers.

Tumors can undergo long periods of dormancy, with cancer cells entering a largely quiescent, nonproliferative state before reactivation and outgrowth. To understand the role of the extracellular matrix (ECM) in regulating tumor dormancy, we created an in vitro cell culture system with carefully controlled ECM substrates to observe entrance into and exit from dormancy with live imaging. We saw that cell populations capable of surviving entrance into long-term dormancy were heterogeneous, containing quiescent, cell cycle-arrested, and actively proliferating cells. Cell populations capable of entering dormancy formed an organized, fibrillar fibronectin matrix via α v β 3 and α 5 β 1 integrin adhesion, ROCK-generated tension, and TGFβ2 stimulation, and cancer cell outgrowth after dormancy required MMP-2-mediated fibronectin degradation. We propose this approach as a useful, in vitro method to study factors important in regulating dormancy, and we used it here to elucidate a role for fibronectin deposition and MMP activation., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)

In pepper crops, rootstocks that tolerate salt stress are not used because available commercial rootstocks offer limited profits. In this context, we obtained the hybrid NIBER®, a new salinity-tolerant rootstock that has been tested under real salinity field conditions for 3 years with 32%-80% higher yields than ungrafted pepper plants. This study aimed to set up the initial mechanisms involved in the salinity tolerance of grafted pepper plants using NIBER® as a rootstock to study root-shoot behavior, a basic requirement to develop efficient rootstocks. Gas exchange, Na + /K + , antioxidant capacity, nitrate reductase activity, ABA, proline, H 2 O 2 , phenols, MDA concentration and biomass were measured in ungrafted plants of cultivar Adige (A), self-grafted (A/A), grafted onto NIBER® (A/N) and reciprocal grafted plants (N/A), all exposed to 0 mM and 70 mM NaCl over a 10-day period. Salinity significantly and quickly decreased photosynthesis, stomatal conductance and nitrate reductase activity, but to lower extent in A/N plants compared to A, A/A and N/A. A/N plants showed decreases in the Na + /K + ratio, ABA content and lipid peroxidation activity. This oxidative damage alleviation in A/N was probably due to an enhanced H 2 O 2 level that activates antioxidant capacity to cope salinity stress, and acts as a signal molecule rather than a damaging one by contributing a major increase in phenols and, to a lesser extent, in proline concentration. These traits led to a minor impact on biomass in A/N plants under salinity conditions. Only the plants with the NIBER® rootstock controlled the scion by modulating responses to salinity., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Bioengineers have designed numerous instructive brain extracellular matrix (ECM) environments with tailored and tunable protein compositions and biomechanical properties in vitro to study astrocyte reactivity during trauma and inflammation. However, a major limitation of both protein-based and synthetic model microenvironments is that astrocytes within fail to retain their characteristic stellate morphology and quiescent state without becoming activated under "normal" culture conditions. Here, a synthetic hydrogel is introduced, which for the first time demonstrates maintenance of astrocyte quiescence and activation on demand. With this synthetic brain hydrogel, the brain-specific integrin-binding and matrix metalloprotease-degradable domains of proteins are shown to control astrocyte star-shaped morphologies, and an ECM condition that maintains astrocyte quiescence with minimal activation can be achieved. In addition, activation can be induced in a dose-dependent manner via both defined cytokine cocktails and low molecular weight hyaluronic acid. This synthetic brain hydrogel is envisioned as a new tool to study the physiological role of astrocytes in health and disease., (© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Cell motility is a critical aspect of several processes, such as wound healing and immunity; however, it is dysregulated in cancer. Current limitations of imaging tools make it difficult to study cell migration in vivo . To overcome this, and to identify drivers from the microenvironment that regulate cell migration, bioengineers have developed 2D (two-dimensional) and 3D (three-dimensional) tissue model systems in which to study cell motility in vitro , with the aim of mimicking elements of the environments in which cells move in vivo . However, there has been no systematic study to explicitly relate and compare cell motility measurements between these geometries or systems. Here, we provide such analysis on our own data, as well as across data in existing literature to understand whether, and which, metrics are conserved across systems. To our surprise, only one metric of cell movement on 2D surfaces significantly and positively correlates with cell migration in 3D environments (percent migrating cells), and cell invasion in 3D has a weak, negative correlation with glioblastoma invasion in vivo . Finally, to compare across complex model systems, in vivo data, and data from different labs, we suggest that groups report an effect size, a statistical tool that is most translatable across experiments and labs, when conducting experiments that affect cellular motility., (© 2019 The Authors. Bioengineering & Translational Medicine published by Wiley Periodicals, Inc. on behalf of The American Institute of Chemical Engineers.)

Bioengineers have built models of the tumour microenvironment (TME) in which to study cell-cell interactions, mechanisms of cancer growth and metastasis, and to test new therapies. These models allow researchers to culture cells in conditions that include features of the in vivo TME implicated in regulating cancer progression, such as extracellular matrix (ECM) stiffness, integrin binding to the ECM, immune and stromal cells, growth factor and cytokine depots, and a three-dimensional geometry more representative of the in vivo TME than tissue culture polystyrene (TCPS). These biomaterials could be particularly useful for drug screening applications to make better predictions of efficacy, offering better translation to preclinical models and clinical trials. However, it can be challenging to compare drug response reports across different biomaterial platforms in the current literature. This is, in part, a result of inconsistent reporting and improper use of drug response metrics, and vast differences in cell growth rates across a large variety of biomaterial designs. This study attempts to clarify the definitions of drug response measurements used in the field, and presents examples in which these measurements can and cannot be applied. We suggest as best practice to measure the growth rate of cells in the absence of drug, and follow our 'decision tree' when reporting drug response metrics. This article is part of a discussion meeting issue 'Forces in cancer: interdisciplinary approaches in tumour mechanobiology'.

In vegetables, tolerance to drought can be improved by grafting commercial varieties onto drought tolerant rootstocks. Grafting has emerged as a tool that copes with drought stress. In previous results, the A25 pepper rootstock accession showed good tolerance to drought in fruit production terms compared with non-grafted plants and other rootstocks. The aim of this work was to study if short-term exposure to drought in grafted plants using A25 as a rootstock would show tolerance to drought now. To fulfill this objective, some physiological processes involved in roots (rootstock) and leaves (scion) of grafted pepper plants were analyzed. Pepper plants not grafted (A), self-grafted (A/A), and grafted onto a tolerant pepper rootstock A25 (A/A25) were grown under severe water stress induced by PEG addition (-0.55 MPa) or under control conditions for 7 days in hydroponic pure solution. According to our results, water stress severity was alleviated by using the A25 rootstock in grafted plants (A/A25), which indicated that mechanisms stimulated by roots are essential to withstand stress. A/A25 had a bigger root biomass compared with plants A and A/A that resulted in better water absorption, water retention capacity and a sustained CO 2 assimilation rate. Consequently, plants A/A25 had a better carbon balance, supported by greater nitrate reductase activity located mainly in leaves. In the non-grafted and self-grafted plants, the photosynthesis rate lowered due to stomatal closure, which limited transpiration. Consequently, part of NO 3 - uptake was reduced in roots. This condition limited water uptake and CO 2 fixation in plants A and A/A under drought stress, and accelerated oxidative damage by producing reactive oxygen species (ROS) and H 2 O 2 , which were highest in their leaves, indicating great sensitivity to drought stress and induced membrane lipid peroxidation. However, drought deleterious effects were slightly marked in plants A compared to A/A. To conclude, the A25 rootstock protects the scion against oxidative stress, which is provoked by drought, and shows better C and N balances that enabled the biomass to be maintained under water stress for short-term exposure, with higher yields in the field.

Michael-type addition reactions are widely used to polymerize biocompatible hydrogels. The thiol-maleimide modality achieves the highest macromer coupling efficiency of the reported Michael-type pairs, but the resulting hydrogel networks are heterogeneous because polymerization is faster than the individual components can be manually mixed. The reactivity of the thiol dictates the overall reaction speed, which can be slowed in organic solvents and acidic buffers. Since these modifications also reduce the biocompatibility of resulting hydrogels, we investigated a series of biocompatible buffers and crosslinkers to decelerate gelation while maintaining high cell viability. We found that lowering the polymer weight percentage (wt%), buffer concentration, and pH slowed gelation kinetics, but crosslinking with an electronegative peptide was optimal for both kinetics and cell viability. Including a high glucose medium supplement in the polymer solvent buffer improved the viability of the cells being encapsulated without impacting gelation time. Slowing the speed of polymerization resulted in more uniform hydrogels, both in terms of visual inspection and the diffusion of small molecules through the network. However, reactions that were too slow resulted in non-uniform particle dispersion due to settling, thus there is a trade-off in hydrogel network uniformity versus cell distribution in the hydrogels when using these networks in cell applications., Statement of Significance: The polymer network of thiol-maleimide hydrogels assembles faster than individual components can be uniformly mixed due to their fast gelation kinetics. The lack of homogeneity can result in variable cell-based assay results, resulting in batch-to-batch variability and limiting their use in predictive screening assays. Although these hydrogels are incredibly useful in tissue engineering, this network heterogeneity is a known problem in the field. We screened a variety of possible techniques to slow down the reaction speed and improve the homogeneity of these hydrogels, without sacrificing the viability and distribution of encapsulated cells. As others have reported, an electronegative crosslinker was the most effective technique to slow the reaction, but the chemical modification required is technically challenging. Of interest to a broad community, we screened buffer type, strength, and crosslinker electronegativity to find an optimal reaction speed that allows for high cell viability and small molecule diffusion, without allowing cells to settle during gelation, allowing application of these materials to the drug screening industry and tissue engineering community., (Copyright © 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

ECM stiffness is emerging as a prognostic marker of tumor aggression or potential for relapse. However, conflicting reports muddle the question of whether increasing or decreasing stiffness is associated with aggressive disease. This chapter discusses this controversy in more detail, but the fact that tumor stiffening plays a key role in cancer progression and in regulating cancer cell behaviors is clear. The impact of having in vitro biomaterial systems that could capture this stiffening during tumor evolution is very high. These cell culture platforms could help reveal the mechanistic underpinnings of this evolution, find new therapeutic targets to inhibit the cross talk between tumor development and ECM stiffening, and serve as better, more physiologically relevant platforms for drug screening.

The performance of a salt-tolerant pepper (Capsicum annuum L.) accession (A25) utilized as a rootstock was assessed in two experiments. In a first field experiment under natural salinity conditions, we observed a larger amount of marketable fruit (+75%) and lower Blossom-end Root incidence (-31%) in commercial pepper cultivar Adige (A) grafted onto A25 (A/A25) when compared with ungrafted plants. In order to understand this behavior a second greenhouse experiment was conducted to determine growth, mineral partitioning, gas exchange and chlorophyll a fluorescence parameters, antioxidant systems and proline content in A and A/A25 plants under salinity conditions (80 mM NaCl for 14 days). Salt stress induced significantly stunted growth of A plants (-40.6% of leaf dry weight) compared to the control conditions, while no alterations were observed in A/A25 at the end of the experiment. Accumulation of Na(+) and Cl(-) in leaves and roots was similar in either grafted or ungrafted plants. Despite the activation of protective mechanisms (increment of superoxide dismutase, catalase, ascorbate peroxidase activity and non-photochemical quenching), A plants showed severely reduced photosynthetic CO2 assimilation (-45.6% of AN390) and substantial buildup of malondialdehyde (MDA) by-product, suggesting the inability to counteract salt-triggered damage. In contrast, A/A25 plants, which had a constitutive enhanced root apparatus, were able to maintain the shoot and root growth under salinity conditions by supporting the maintained photosynthetic performance. No increases in catalase and ascorbate peroxidase activities were observed in response to salinity, and MDA levels increased only slightly; indicating that alleviation of oxidative stress did not occur in A/A25 plants. In these plants the increased proline levels could protect enzymatic stability from salt-triggered damage, preserving the photosynthetic performance. The results could indicate that salt stress was vanished by the lack of negative effects on photosynthesis that support the maintained plant growth and increased marketable yield of the grafted plants., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Grafting has been proposed as an interesting strategy that improves the responses of crops under salinity. In pepper, we reported increased fruit yield of the commercial 'Adige' cultivar under salinity when grafted onto accessions Capsicum chinense Jacq. 'ECU-973' (12) and Capsicum baccatum L. var. pendulum 'BOL-58' (14), whereas no effect was observed when grafted onto accession Capsicum annuum L var. 'Serrano' (5). We also analysed the physiological and biochemical mechanisms related to the tolerance conferred by these rootstocks. Responses to salinity (40 mM NaCl) were studied in the different plant combinations for 30 days by determining water relations, mineral content, proline accumulation, photosynthetic parameters, nitrate reductase activity and antioxidant capacity. Higher salt tolerance was achieved when the 'Adige' cultivar was grafted onto the 12 genotype, which allowed not only lower Na(+) and Cl(-) accumulation in the scion, but also ion selectivity maintenance, particularly Na(+)/K(+) discrimination. These traits led to a minor negative impact on photosynthesis, nitrate reductase activity and lipid peroxidation in grafted scion leaves. This work suggests that using tolerant pepper rootstocks that maintain the scion's ion homeostasis is a promising strategy to provide salinity tolerance and can consequently improve crop yield., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Recent studies have shown that tolerance to abiotic stress, including water stress, is improved by grafting. In a previous work, we took advantage of the natural variability of Capsicum spp. and selected accessions tolerant and sensitive to water stress as rootstocks. The behavior of commercial cultivar 'Verset' seedlings grafted onto the selected rootstocks at two levels of water stress provoked by adding 3.5 and 7% PEG (polyethylene glycol) was examined over 14 days. The objective was to identify the physiological traits responsible for the tolerance provided by the rootstock in order to determine if the tolerance is based on the maintenance of the water relations under water stress or through the activation of protective mechanisms. To achieve this goal, various physiological parameters were measured, including: water relations; proline accumulation; gas exchange; chlorophyll fluorescence; nitrate reductase activity; and antioxidant capacity. Our results indicate that the effect of water stress on the measured parameters depends on the duration and intensity of the stress level, as well as the rootstock used. Under control conditions (0% PEG) all plant combinations showed similar values for all measured parameters. In general terms, PEG provoked a strong decrease in the gas exchange parameters in the cultivar grafted onto the sensitive accessions, as also observed in the ungrafted plants. This effect was related to lower relative water content in the plants, provoked by an inefficient osmotic adjustment that was dependent on reduced proline accumulation. At the end of the experiment, chronic photoinhibition was observed in these plants. However, the plants grafted onto the tolerant rootstocks, despite the reduction in photosynthetic rate, maintained the protective capacity of the photosynthetic machinery mediated by osmotic adjustment (based on higher proline content). In addition, water stress limited uptake and further NO3(-) transfer to the leaves. Increased nitrate reductase activity in the roots was observed, mainly in plants grafted onto the sensitive rootstocks, as well as the ungrafted plants, and this was associated with the lessened flux to the leaves. This study suggests that PEG-induced water stress can be partially alleviated by using tolerant accessions as rootstocks., (Copyright © 2014 Elsevier GmbH. All rights reserved.)