פרסומים

Abstract: Development of mechanically stable and multifunctional biomaterials for sensing, repair, and regeneration applications is of great importance. Herein, we investigate the potential of recombinant resilin-like (Res) nanocomposite elastomer as a template biomaterial for regenerative devices such as adhesive bandages or films, electrospun fibers, screws, sutures, and drug delivery vehicles. Exon I (Rec1) from the native resilin gene of Drosophila (CG15920) was fused with collagen-binding domain (ColBD) from Clostridium histolyticum and expressed in Komagataella pastoris (formerly Pichia pastoris). The 100% binding of Resilin-ColBD (Res-ColBD) to collagen I was shown at a 1:1 ratio by mass. Atomic force microscopy results in force mode show a bimodal profile for the ColBD-binding interactions. Moreover, based on the force-volume map, Res-ColBD adhesion to collagen was statistically significantly higher than resilin without ColBD. Lay Summary: Designing advanced biomaterials that will not only withstand the repetitive mechanical loading and flexibility of tissues but also retain biochemical and biophysical interactions remains challenging. The combination of physical, biological, and chemical cues is vital for disease regulation, healing, and ultimately complete regeneration of functional human tissues. Resilin is a super elastic and highly resilient natural protein with good biocompatibility but lacks specific biological and chemical cues. Therefore, resilin decorated with collagen I–binding domain is proposed as a functional nanocomposite template biomaterial. Collagen I is an ideal binding target, as it is the most abundant structural protein found in human body including scars that affect unwanted adhesion. Future Work: Musculoskeletal-related injuries and disorders are the second largest cause of disabilities worldwide. Significant pain, neurological discomfort, limited mobility, and substantial financial burden are associated with these disorders. Thus, biocompatible materials comprised of resilin with collagen-binding domain, such as films adhesive bandages (films, fiber matts, or hydrogels), sutures, screws and rods, three-dimensional scaffolds, and delivery vehicles, will be designed and evaluated for multiple musculoskeletal-related regeneration applications. © 2019, The Regenerative Engineering Society.

Abstract: We postulate that the extensive cell and tissue damage inflicted by many infectious, inflammatory and post-inflammatory episodes is an enled result of a synergism among the invading microbial agents, host neutrophils and dead and dying cells in the nidus. Microbial toxins and other metabolites along with the plethora of pro-inflammatory agents released from activated neutrophils massively recruited to the infectious sites and high levels of cationic histones, other cationic peptides, proteinases and Th1 cytokines released from activated polymorphonuclear neutrophils (PMNs) and from necrotized tissues may act in concert (synergism) to bring about cell killing and tissue destruction. Multiple, diverse interactions among the many potential pro-inflammatory moieties have been described in these complex lesions. Such infections are often seen in the skin and aerodigestive tract where the tissue is exposed to the environment, but can occur in any tissue. Commonly, the tissue-destructive infections are caused by group A streptococci, pneumococci, Staphylococcus aureus, meningococci, Escherichia coli and Shigella, although many other microbial species are seen on occasion. All these microbial agents are characterized by their ability to recruit large numbers of PMNs. Given the complex nature of the disease process, it is proposed that, to treat these multifactorial disorders, a “cocktail” of anti-inflammatory agents combined with non-bacteriolytic antibiotics and measures to counteract the critical toxic role of cationic moieties might prove more effective than a strategy based on attacking the bacteria alone.

A new single-step proline-potassium acetate promoted reductive dehydroxylation of α-ketols is reported. We introduce the unexplored reactivity of proline and, for the first time, reveal its ability to function as a reducing agent. The developed metal-free and open-flask operation generally results in good yields. Our protocol allows the challenging selective dehydroxylation of hydroxyketones without affecting other functional groups. This journal is © The Royal Society of Chemistry.

Carbamazepine (CBZ)is an anticonvulsant drug used for epilepsy and other disorders. Prescription of CBZ during pregnancy increases the risk for congenital malformations. CBZ is ubiquitous in effluents and persistent during wastewater treatment. Thus, it is re-introduced into agricultural ecosystems upon irrigation with reclaimed wastewater. People consuming produce irrigated with reclaimed wastewater were found to be exposed to CBZ. However, environmental concentrations of CBZ (μg L−1)are magnitudes lower than its therapeutic levels (μg ml−1), raising the question of whether and how environmental levels of CBZ affect embryonic development. The chick embryo is a powerful and highly sensitive amniotic model system that enables to assess environmental contaminants in the living organism. Since the chick embryonic development is highly similar to mammalians, yet, it develops in an egg, toxic effects can be directly analyzed in a well-controlled system without maternal influences. This research utilized the chick embryo to test whether CBZ is embryo-toxic by using morphological, cellular, molecular and imaging strategies. Three key embryonic stages were monitored: after blastulation (st.1HH), gastrulation/neurulation (st.8HH)and organogenesis (st.15HH). Here we demonstrate that environmental relevant concentrations of CBZ impair morphogenesis in a dose- and stage- dependent manner. Effects on gastrulation, neural tube closure, differentiation and proliferation were exhibited in early stages by exposing embryos to CBZ dose as low as 0.1 μg L−1. Quantification of developmental progression revealed a significant difference in the total score obtained by CBZ-treated embryos compared to controls (up to 5-fold difference, p < 0.05). Yet, defects were unnoticed as embryos passed gastrulation/neurulation. This study provides the first evidence for teratogenic effect of environmental-relevant concentrations of CBZ in amniotic embryos that impair early but not late stages of development. These findings call for in-depth risk analysis to ensure that the environmental presence of CBZ and other drugs is not causing irreversible ecological and public-health damages. © 2019

Carbamazepine (CBZ)is an anticonvulsant drug used for epilepsy and other disorders. Prescription of CBZ during pregnancy increases the risk for congenital malformations. CBZ is ubiquitous in effluents and persistent during wastewater treatment. Thus, it is re-introduced into agricultural ecosystems upon irrigation with reclaimed wastewater. People consuming produce irrigated with reclaimed wastewater were found to be exposed to CBZ. However, environmental concentrations of CBZ (μg L−1)are magnitudes lower than its therapeutic levels (μg ml−1), raising the question of whether and how environmental levels of CBZ affect embryonic development. The chick embryo is a powerful and highly sensitive amniotic model system that enables to assess environmental contaminants in the living organism. Since the chick embryonic development is highly similar to mammalians, yet, it develops in an egg, toxic effects can be directly analyzed in a well-controlled system without maternal influences. This research utilized the chick embryo to test whether CBZ is embryo-toxic by using morphological, cellular, molecular and imaging strategies. Three key embryonic stages were monitored: after blastulation (st.1HH), gastrulation/neurulation (st.8HH)and organogenesis (st.15HH). Here we demonstrate that environmental relevant concentrations of CBZ impair morphogenesis in a dose- and stage- dependent manner. Effects on gastrulation, neural tube closure, differentiation and proliferation were exhibited in early stages by exposing embryos to CBZ dose as low as 0.1 μg L−1. Quantification of developmental progression revealed a significant difference in the total score obtained by CBZ-treated embryos compared to controls (up to 5-fold difference, p < 0.05). Yet, defects were unnoticed as embryos passed gastrulation/neurulation. This study provides the first evidence for teratogenic effect of environmental-relevant concentrations of CBZ in amniotic embryos that impair early but not late stages of development. These findings call for in-depth risk analysis to ensure that the environmental presence of CBZ and other drugs is not causing irreversible ecological and public-health damages.

In recent decades, anthropogenic activity and climate changes have reshaped global weed dispersal and establishment in new territories. This study aimed to evaluate the effectiveness of propane flaming approach in the control of perennial invasive and native Mediterranean broadleaf and grass weeds. The invasive weeds, Cyperus rotundus, Sorghum halepense, and Ecballium elaterium, were treated multiple times with a single propane dose (2.5 kg propane km −1 ), using the broadcast technique. The local annual weeds, Sinapis arvensis, Lavatera trimestris, and Avena sativa, were treated once at five propane doses (0–2.5 kg propane km −1 ), using the cross-row technique. Dose-response analysis was performed. Three applications provided effective control (up to >90%) for all tested perennials, and affected seed and flower production in Sorghum halepense and Ecballium elaterium, respectively. However, the timing of the sequential application had a significant impact on the degree of control, in terms of dry weight reduction and seed production. Weed density had an impact on control efficacy but was only a significant determinant for Ecballium elaterium. Cross-row application was effective during early growth stages of broadleaf weeds (ED 50 < 1.2 kg propane km −1 ), but was less effective during later growth stages (ED 50 > 2.6 kg propane km −1 ). For grass weeds, both early and late application were ineffective (ED 50 > 4.1 kg propane km −1 ). More research is needed to optimize this weed control tactic for various cropping systems and weed species. Implementation of this novel approach into integrated weed management programs will increase the control efficacy of invasive weed under the projected climate changes and reduce the evolution of herbicide-resistant weeds. © 2019 by the authors.

Abstract Background Urethral obstruction (UO) is a common complication of feline idiopathic cystitis (FIC). Robust treatment recommendations to prevent its recurrence are scarce. Objectives To evaluate meloxicam treatment for prevention of clinical recrudescence in male cats with obstructive FIC. Animals Fifty-one client-owned cats. Methods Prospective, randomized clinical trial. Every male cat with FIC-associated UO was deemed eligible for the study and was recruited during hospitalization. After discharge, cats were treated with phenoxybenzamine and alprazolam for 2?weeks, with (24 cats) or without (27 cats) low-dose meloxicam (0.025?mg/kg/day PO) and monitored for 6?months. Results Cumulative number (%) of cats with recurrent UO at 10?days, 1-, 2-, and 6-months after discharge was 1 (2%), 2 (4%), 4 (8%), and 8 (16%), respectively. Overall, 12 (24%) cats experienced signs of recurrent FIC within 6?months, with (8 cats) or without (4 cats) concurrent UO. No difference in the cumulative incidence of UO within 6?months was detected with addition of meloxicam (odds ratio [95% confidence interval], 0.63 [0.13-2.97]; P =?.70). All cats were alive at 6?months. Conclusions and Clinical Importance No clinical benefit was detected with the addition of low-dose meloxicam to phenoxybenzamine and alprazolam treatment for 2?weeks after discharge. Nevertheless, this study was underpowered to identify potential differences, and its findings must be corroborated in larger studies.

BACKGROUND: Bone marrow-derived mesenchymal stem cell (BMSC) transplantation is one of the new therapeutic strategies for treating ischemic brain and heart tissues. However, the poor survival rate of transplanted BMSCs in ischemic tissue, due to high levels of reactive oxygen species (ROS), limits the therapeutic efficacy of this approach. Considering that BMSC survival may greatly enhance the effectiveness of transplantation therapy, development of effective therapeutics capable of mitigating oxidative stress-induced BMSC apoptosis is an important unmet clinical need. METHODS: BMSCs were isolated from the 4-week-old male Sprague Dawley rats by whole bone marrow adherent culturing, and the characteristics were verified by morphology, immunophenotype, adipogenic, and osteogenic differentiation potential. BMSCs were pretreated with artemisinin, and H(2)O(2) was used to induce apoptosis. Cell viability was detected by MTT, FACS, LDH, and Hoechst 33342 staining assays. Mitochondrial membrane potential ($Δ$$\Psi$m) was measured by JC-1 assay. The apoptosis was analyzed by Annexin V-FITC/PI and Caspase 3 Activity Assay kits. ROS level was evaluated by using CellROX® Deep Red Reagent. SOD, CAT, and GPx enzymatic activities were assessed separately using Cu/Zn-SOD and Mn-SOD Assay Kit with WST-8, Catalase Assay Kit, and Total Glutathione Peroxidase Assay Kit. The effects of artemisinin on protein expression of BMSCs including p-Erk1/2, t-Erk1/2, p-c-Raf, p-p90(rsk), p-CREB, BCL-2, Bax, p-Akt, t-Akt, $\beta$-actin, and GAPDH were measured by western blotting. RESULTS: We characterized for the first time the protective effect of artemisinin, an anti-malaria drug, using oxidative stress-induced apoptosis in vitro, in rat BMSC cultures. We found that artemisinin, at clinically relevant concentrations, improved BMSC survival by reduction of ROS production, increase of antioxidant enzyme activities including SOD, CAT, and GPx, in correlation with decreased Caspase 3 activation, lactate dehydrogenase (LDH) release and apoptosis, all induced by H(2)O(2). Artemisinin significantly increased extracellular-signal-regulated kinase 1/2 (Erk1/2) phosphorylation, in a concentration- and time-dependent manner. PD98059, the specific inhibitor of the Erk1/2 pathway, blocked Erk1/2 phosphorylation and artemisinin protection. Similarly, decreased expression of Erk1/2 by siRNA attenuated the protective effect of artemisinin. Additionally, when the upstream activator KRAS was knocked down by siRNA, the protective effect of artemisinin was also blocked. These data strongly indicated the involvement of the Erk1/2 pathway. Consistent with this hypothesis, artemisinin increased the phosphorylation of Erk1/2 upstream kinases proto-oncogene c-RAF serine/threonine-protein kinase (c-Raf) and of Erk1/2 downstream targets p90 ribosomal s6 kinase (p90(rsk)) and cAMP response element binding protein (CREB). In addition, we found that the expression of anti-apoptotic protein B cell lymphoma 2 protein (BcL-2) was also upregulated by artemisinin. CONCLUSION: These studies demonstrate the proof of concept of artemisinin therapeutic potential to improve survival in vitro of BMSCs exposed to ROS-induced apoptosis and suggest that artemisinin-mediated protection occurs via the activation of c-Raf-Erk1/2-p90(rsk)-CREB signaling pathway.

The protein–water–ice contact angle is a controlling parameter in diverse fields. Here we show that data from three different experiments, at three different length scales, with three different proteins, in three different laboratories yield a consistent value for the protein–water–ice contact angle (88.0 ± 1.3°) when analyzed using the Gibbs–Thomson equation. The measurements reinforce the validity of each other, and the fact that similar values are obtained across diverse length scales, experiments, and proteins yields insight into protein–water interactions and the applicability of thermodynamics at the nanoscale.The protein–water–ice contact angle is a controlling parameter in diverse fields. Here we show that data from three different experiments, at three different length scales, with three different proteins, in three different laboratories yield a consistent value for the protein–water–ice contact angle (88.0 ± 1.3°) when analyzed using the Gibbs–Thomson equation. The measurements reinforce the validity of each other, and the fact that similar values are obtained across diverse length scales, experiments, and proteins yields insight into protein–water interactions and the applicability of thermodynamics at the nanoscale.

BACKGROUND: Fig fruit are highly perishable at the tree-ripe (TR) stage. Commercial-ripe (CR) fruit, which are harvested before the TR stage for their postharvest transportability and shelf-life advantage, are inferior to TR fruit in size, color and sugar content. The succulent urn-shaped receptacle, serving as the protective structure and edible part of the fruit, determines fruit quality. Quantitative iTRAQ and RNA-Seq were performed to reveal the differential proteomic and transcriptomic traits of the receptacle at the two harvest stages. RESULTS: We identified 1226 proteins, of which 84 differentially abundant proteins (DAPs) were recruited by criteria of abundance fold-change (FC) ≥1.3 and p < 0.05 in the TR/CR receptacle proteomic analysis. In addition, 2087 differentially expressed genes (DEGs) were screened by ≥2-fold expression change: 1274 were upregulated and 813 were downregulated in the TR vs. CR transcriptomic analysis. Ficin was the most abundant soluble protein in the fig receptacle. Sucrose synthase, sucrose-phosphate synthase and hexokinase were all actively upregulated at both the protein and transcriptional levels. Endoglucanase, expansin, beta-galactosidase, pectin esterase and aquaporins were upregulated from the CR to TR stage at the protein level. In hormonal synthesis and signaling pathways, high protein and transcriptional levels of aminocyclopropane-1-carboxylate oxidase were identified, together with a few diversely expressed ethylene-response factors, indicating the potential leading role of ethylene in the ripening process of fig receptacle, which has been recently reported as a non-climacteric tissue. CONCLUSIONS: We present the first delineation of intra- and inter-omic changes in the expression of specific proteins and genes of TR vs. CR fig receptacle, providing valuable candidates for further study of fruit-quality formation control and fig cultivar innovation to accommodate market demand.