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In recent years, the rapid development of genetically modified (GM) technology has raised concerns about the safety of GM crops and foods for human health and the ecological environment. Gene flow from GM crops to other crops, especially in the Brassicaceae family, might pose a threat to the environment due to their weediness. Hence, finding reliable, quick, and low-cost methods to detect and monitor the presence of GM crops and crop products is important. In this study, we used visible near-infrared (Vis-NIR) spectroscopy for the effective discrimination of GM and non-GM Brassica napus, B. rapa, and F1 hybrids (B. rapa X GM B. napus). Initially, Vis-NIR spectra were collected from the plants, and the spectra were preprocessed. A combination of different preprocessing methods (four methods) and various modeling approaches (eight methods) was used for effective discrimination. Among the different combinations, the Savitzky-Golay and Support Vector Machine combination was found to be an optimal model in the discrimination of GM, non-GM, and hybrid plants with the highest accuracy rate (100%). The use of a Convolutional Neural Network with Normalization resulted in 98.9%. The same higher accuracy was found in the use of Gradient Boosted Trees and Fast Large Margin approaches. Later, phenolic acid concentration among the different plants was assessed using GC-MS analysis. Partial least squares regression analysis of Vis-NIR spectra and biochemical characteristics showed significant correlations in their respective changes. The results showed that handheld Vis-NIR spectroscopy combined with chemometric analyses could be used for the effective discrimination of GM and non-GM B. napus, B. rapa, and F1 hybrids. Biochemical composition analysis can also be combined with the Vis-NIR spectra for efficient discrimination.

Penthorum chinense Pursh is a plant belonging to the Crassulaceae family and is a representative perennial herb that grows nationwide in watersides and wetlands. This study was conducted to enhance the competitiveness of wild plant seeds and contribute to the conservation of species diversity through the evaluation of antioxidant and antidiabetic activities of P. chinense seed extracts based on extraction solvents. The total phenolics content was 19.83 mg GAE/g seeds in the 75% ethanol extract, whereas the flavonoid content was the highest (10.89 mg QE/g seeds) in the 75% methanol extract. Total phenolicss content was higher in the organic solvent extract than in the water extract. The DPPH radical scavenging activity was the highest (IC50 27.60 μg/mL) in the 75% methanol extract, and the ABTS radical scavenging activity was the highest (IC50 53.54 μg/mL) in the 75% methanol extract. Total antioxidant capacity was the highest (30.87 μmol AAE/g) in the 75% ethanol extract. The α-glucosidase-inhibitory activity was the highest (IC50 46.47 μg/mL) in the 75% ethanol extract. From the results of this study, it was confirmed that P. chinense seed extracts, which possess high phenolics content, exhibit antioxidant and antidiabetic effects, and could be used as an effective health functional food material and natural medicine.

A highly dispersed NiMoP phosphide catalyst with an average particle size of 9.1 nm and a carbon content of 53.5 wt% was firstly synthesized by a carbothermal route using glucose as a carbon source. The current NiMoP catalyst can show higher catalytic activity and stability compared to the one prepared by a traditional H2 reduction route.

Visually significant corneal injuries and subsequent scarring collectively represent a major global human health challenge, affecting millions of people worldwide. Unfortunately, less than 2% of patients who could benefit from a sight-restoring corneal transplant have access to cadaveric donor corneal tissue. Thus, there is a critical need for new ways to repair corneal defects that drive proper epithelialization and stromal remodeling of the wounded area without the need for cadeveric donor corneas. Emerging therapies to replace the need for donor corneas include pre-formed biosynthetic buttons and in situ-forming matrices that strive to achieve the transparency, biocompatibility, patient comfort, and biointegration that is possible with native tissue. Herein, we report on the development of an in situ-forming hydrogel of collagen type I crosslinked via multi-functional polyethylene glycol (PEG)-N-hydroxysuccinimide (NHS) and characterize its biophysical properties and regenerative capacity both in vitro and in vivo. The hydrogels form under ambient conditions within minutes upon mixing without the need for an external catalyst or trigger such as light or heat, and their transparency, degradability, and stiffness are modulated as a function of number of PEG arms and concentration of PEG. In addition, in situ-forming PEG-collagen hydrogels support the migration and proliferation of corneal epithelial and stromal cells on their surface. In vivo studies in which the hydrogels were formed in situ over stromal keratectomy wounds without sutures showed that they supported multi-layered surface epithelialization. Overall, the in situ forming PEG-collagen hydrogels exhibited physical and biological properties desirable for a corneal stromal defect wound repair matrix that could be applied without the need for sutures or an external trigger such as a catalyst or light energy.

The occurrence of exotic weeds and their influx into farmlands due to climate change poses many problems. Therefore, it is necessary to generate a prediction model for the occurrence pattern of these exotic weeds based on scientific evidence and devise prevention measures. The photosynthetic apparatus is known as the most temperaturesensitive component of a plant cell and its initial response to temperature stress is to inhibit the activation of photosystem II. This study investigated the potential of OJIP transients in assessing temperature stress in exotic weeds. The four exotic weeds currently flowing into Korean farmlands include Amaranthus spinosus, Conyza bonariensis, Crassocephalum crepidioides, and Amaranthus viridis. These weeds were treated at 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, and 40°C and the OJIP curves and JIP parameters were measured and analyzed. The results showed that heat and chilling stress affected the photosystem II (PSII) electron transport of A. spinosus, whereas C. crepidioides and A. viridis were more affected by high-temperature stress than by low-temperature stress. Lastly, C. bonariensis showed resistance to both high and low-temperature stress. The results of this study suggest that OJIP transients and JIP parameters can be used to analyze damage to the photosynthetic apparatus by temperature stress and that they can serve as sensitive indicators for the occurrence pattern of exotic weeds.

This study aimed to determine the type of seed dormancy and to identify a suitable method of dormancy-breaking for an efficient seed viability test of Lysimachia coreana Nakai. To confirm the effect of gibberellic acid (GA3) on seed germination at different temperatures, germination tests were conducted at 5, 15, 20, 25, 20/10, and 25/15 °C (12/12 h, light/dark), using 1% agar with 100, 250, and 500 mg·L−1 GA3. Seeds were also stratified at 5 and 25/15 °C for 6 and 9 weeks, respectively, and then germinated at the same temperature. Seeds treated with GA3 demonstrated an increased germination rate (GR) at all temperatures except 5 °C. The highest GR was 82.0% at 25/15 °C and 250 mg·L−1 GA3 (4.8 times higher than the control (14.0%)). Additionally, GR increased after cold stratification, whereas seeds did not germinate after warm stratification at all temperatures. After cold stratification, the highest GR was 56.0% at 25/15 °C, which was lower than the GR observed after GA3 treatment. We hypothesized that L. coreana seeds have a non-deep physiological dormancy and concluded that 250 mg·L−1 GA3 treatment is more effective than cold stratification (9 weeks) for L. coreana seed-dormancy-breaking.

Platycosides, saponins contained in balloon flower, which have been used as food health supplements for respiratory diseases, have diverse pharmacological effects. Platycosides exhibit better pharmacological activity by hydrolyzing their own sugars. However, to date, there have been no studies on the production of deglucosylated platycodin D suitable for food applications. In this study, Pluszyme 2000P, which was derived from Aspergillus niger, a food-grade microorganism, was used to completely convert platycoside E into deglucosylated platycodin D. For an efficient and economical production of deglucosylated platycodin D, the productivity was improved approximately 2.4 times by application of high hydrostatic pressure and the discarded balloon flower leaf was used as a substrate. As a result, deglucosylated platycodin D was produced with the highest concentration (3.49 mg/mL) and productivity (581.7 mg/L/h) reported so far. Our results contribute to functional saponin production and the related food industries., Platycoside; Pluszyme 2000P; Food additive; Aspergillus niger; Balloon flower; Medicinal herb.

This study investigated the kind of seed dormancy and seed germination of Gentiana&nbsp, triflora var. japonica (Kusn.) H. Hara for developing a seed propagation method. The seeds were collected in October 2020 from plants at Mt. Sobaeksan, Korea. In a water imbibition experiment, seed weights increased by &gt, 101.9% of their initial masses over 12 h. Effects of incubation temperature (5, 15, 20, 25, 15/6, or 25/15 °C), cold stratification period (5 °C, 0, 4, 8, or 12 weeks), and gibberellic acid (GA3, 0, 10, 100, or 1000 mg∙L−1) and potassium nitrate treatment (KNO3, 0, 1000, 2000, or 4000 mg∙L−1) on seed germination were investigated to characterize seed dormancy. These seeds exhibited underdeveloped embryos during seed dispersal. The seeds failed to reach the final germination of 15.0% after treatment at 5, 15, 20, 25, 15/6, or 25/15 °C. After cold stratification for 8 weeks, the germination increased dramatically by &gt, 90.0% compared to that at 0 weeks. After the GA3 treatment, the germination reached &gt, 80.0% within 5 days. The final germination was 90.0% in the 100 mg∙L−1 GA3 treatment group. However, the KNO3 treatment had no effect on seed germination. Therefore, the G. triflora var. japonica seeds exhibited non-deep simple morphophysiological dormancy.

The therapeutic effects of secreted factors (secretome) produced by bone marrow-derived human mesenchymal stem cells (MSCs) were evaluated as a function of their growth in 2D culture conditions and on 3D electrospun fiber scaffolds. Electrospun fiber scaffolds composed of polycaprolactone and gelatin were fabricated to provide a 3D microenvironment for MSCs, and their mechanical properties were optimized to be similar to corneal tissue. The secretome produced by the MSCs cultured on 3D fiber matrices versus 2D culture dishes were analyzed using a Luminex immunoassay, and the secretome of MSCs cultured on the 3D versus 2D substrates showed substantial compositional differences. Concentrations of factors such as HGF and ICAM-1 were increased over 5 times in 3D cultures compared to 2D cultures. In vitro proliferation and scratch-based wound healing assays were performed to compare the effects of the secretome on corneal fibroblast cells (CFCs) when delivered synchronously from co-cultured MSCs through a trans-well co-culture system versus asynchronously after harvesting the factors separately and adding them to the media. Cell viability of CFCs was sustained for 6 days when co-cultured with MSCs seeded on the fibers but decreased with time under other conditions. Scratch assays showed 95% closure at 48 h when CFCs were co-cultured with MSCs seeded on fibers, while the control group only exhibited 50% closure at 48 h. Electrospun fibers seeded with MSCs were then applied to a rabbit corneal organ culture system, and MSCs seeded on fibers promoted faster epithelialization and less scarring. Corneas were fixed and stained for alpha smooth muscle actin (α-SMA), and then analyzed by confocal microscopy. Immunostaining showed that expression of α-SMA was lower in corneas treated with MSCs seeded on fibers, suggesting suppression of myofibroblastic transformation. MSCs cultured on electrospun fibers facilitate wound healing in CFCs and on explanted corneas through differential secretome profiles compared to MSCs cultured on 2D substrates. Future work is merited to further understand the nature and basis of these differences and their effects in animal models. Statement of significance Previous studies have shown that the secretome of bone marrow-derived mesenchymal stem cells (MSC) is promotes corneal wound healing by facilitating improved wound closure rates and reduction of scarring and neovascularization. The present research is significant because it provides evidence for the modulation of the secretome as a function of the MSC culture environment. This leads to differential expression of therapeutic factors secreted, which can impact corneal epithelial and stromal healing after severe injury. In addition, this article shows that co-continuous delivery of the MSC secretome improves cell migration and proliferation over aliquoted delivery, and that MSCs grown on three-dimensional electrospun fiber constructs may provide a favorable microenvironment for cultured MSCs and as a carrier to deliver their secreted factors to the ocular surface.

Solid–liquid separation of similarly sized organic molecules utilizing sorbents offers the potential for new energy-efficient approaches to a number of important industrial separations such as xylenes (C8) separations., Solid–liquid separation of similarly sized organic molecules utilizing sorbents offers the potential for new energy-efficient approaches to a number of important industrial separations such as xylenes (C8) separations. Research on selective C8 sorption has tended to focus upon rigid porous materials such as zeolites and MOFs but has revealed generally weak selectivity that is inconsistent across the range of C8 molecules. Nevertheless, there are a few recent examples of non-porous molecular materials that exhibit relatively high selectivity for p-xylene (pX) from pX/oX, approaching that of the current benchmark pX sorbent, the zeolite H/ZSM-5. Herein, we report that a L-shaped Ag(i) complex, AgLClO4 (M), which crystallizes as a non-porous molecular solid material, offering exceptional performance for pX selectivity across the range of C8 isomers with liquid extraction selectivity values of 24.0, 10.4 and 6.2 vs. oX, eB and mX, respectively. The pX selectivities over oX and eB are among the highest yet reported. Moreover, M also exhibits strong vapor extraction selectivity and can be regenerated by exposure to vacuum drying.

Severe corneal injuries often result in permanent vision loss and remain a clinical challenge. Human bone marrow-derived mesenchymal stem cells (MSCs) and their secreted factors (secretome) have been studied for their antiscarring, anti-inflammatory, and antiangiogeneic properties. We aimed to deliver lyophilized MSC secretome (MSC-S) within a viscoelastic gel composed of hyaluronic acid (HA) and chondroitin sulfate (CS) as a way to enhance corneal re-epithelialization and reduce complications after mechanical and chemical injuries of the cornea. We hypothesized that delivering MSC-S within HA/CS would have improved wound healing effects compared the with either MSC-S or HA/CS alone. The results showed that a once-daily application of MSC-S in HA/CS enhances epithelial cell proliferation and wound healing after injury to the cornea. It also reduced scar formation, neovascularization, and hemorrhage after alkaline corneal burns. We found that combining MSC-S and HA/CS increased the expression of CD44 receptors colocalized with HA, suggesting that the observed therapeutic effects between the MSC-S and HA/CS are in part mediated by CD44 receptor upregulation and activation by HA. The results from this study demonstrate a reproducible and efficient approach for delivering the MSC-S to the ocular surface for treatment of severe corneal injuries. Stem Cells Translational Medicine 2019;8:478–489

Sulfonated ordered mesoporous carbon (SO3H-OMC) solid acid catalysts from sucrose were prepared using hard-template method, and their catalytic performance as well as the deactivation mechanism for esterification of free fatty acids (FFAs) in waste cooking oils (WCOs) were evaluated. Effects of sulfonation time, sulfonation temperature and hard template structure type for the textural properties and acid properties of SO3H-OMC were systematically investigated by N2 adsorption–desorption, FT-IR, NH3-TPD, TEM and strong acid density analysis. The results indicated that, SO3H-OMC(s)-6-160 catalyst, which was prepared by using SBA-15 as hard template at sulfonation time of 6 h and sulfonation temperature of 160 °C, had well-ordered mesoporous structure and high –SO3H groups density (2.32 mmol g−1). Compared with SO3H-APC-6-160 catalyst, cation-exchange resin D072 and SO3H-OMC(k)-6-160 catalyst, it was found that the SO3H-OMC(s)-6-160 catalyst exhibited highest activity (FFAs conversion was 93.8%) and good stability for the FFAs esterification, attributed to its 2D-hexagonal channels and hydrophobic surface. The –SO3H groups being leached out of SO3H-OMC catalysts into the liquid phase (especially methanol) would be the main reason causing catalyst deactivation.

Large abrasions and deeper ulcers of the cornea can lead to corneal scarring, ulceration and thinning if not promptly and adequately treated. Hyaluronic acid (HA) has been investigated for the treatment of corneal wounds due to its remarkable biocompatibility, transparency and mucoadhesive properties. However, intact linear HA has low retention time on the cornea and chemical crosslinkers to synthesize HA hydrogels can cause toxicity limiting their clinical ocular applications. Here, we used supramolecular HA hydrogels formed by non-covalent host-guest interactions between HA-cyclodextrin and HA-adamantane to evaluate the impact of the hydrogels on corneal wound healing. Supramolecular HA hydrogels facilitated adhesion and spreading of encapsulated human corneal epithelial cells ex vivo and improved corneal wound healing in vivo as an in situ-formed, acellular therapeutic membrane. The HA hydrogels were absorbed within the corneal stroma over time, modulated mesenchymal cornea stromal cell secretome production, reduced cellularity and inflammation of the anterior stroma, and significantly mitigated corneal edema compared to treatment with linear HA and untreated control eyes. Taken together, our results demonstrate supramolecular HA hydrogels as a promising and versatile biomaterial platform for corneal wound healing.

Platycosides, saponins contained in balloon flower, which have been used as food health supplements for respiratory diseases, have diverse pharmacological effects. Platycosides exhibit better pharmacological activity by hydrolyzing their own sugars. However, to date, there have been no studies on the production of deglucosylated platycodin D suitable for food applications. In this study, Pluszyme 2000P, which was derived from Aspergillus niger, a food-grade microorganism, was used to completely convert platycoside E into deglucosylated platycodin D. For an efficient and economical production of deglucosylated platycodin D, the productivity was improved by application of high hydrostatic pressure and the discarded balloon flower leaf was used as a substrate. As a result, deglucosylated platycodin D was produced with the highest concentration and productivity reported so far. Our results contribute to functional saponin production and the related food industries.

Background aims Corneal inflammation after alkali burns often results in vision loss due to corneal opacification and neovascularization. Mesenchymal stem cells (MSCs) and their secreted factors (secretome) have been studied for their anti-inflammatory and anti-angiogenic properties with encouraging results. However, topical instillation of MSCs or their secretome is often accompanied by issues related to delivery or rapid washout. Polyethylene glycol (PEG) and collagen are well-known biomaterials used extensively in scaffolds for tissue engineering. To effectively suppress alkaline burn-induced corneal injury, the authors proposed encapsulating MSCs within collagen gels cross-linked with multi-functional PEG-succinimidyl esters as a means to deliver the secretome of immobilized MSCs. Methods Human MSCs were added to a neutralized collagen solution and mixed with a solution of four-arm PEG-N-hydroxysuccinimide. An ex vivo organ culture was conducted using rabbit corneas injured by alkali burn. MSCs were encapsulated within PEG-collagen hydrogels and injected onto the wounded cornea immediately following alkali burn and washing. Photographs of the ocular surface were taken over a period of 7 days after the alkali burn and processed for immunohistochemical evaluation. Samples were split into three groups: injury without treatment, MSCs alone, and MSCs encapsulated within PEG-collagen hydrogels. Results All corneas in ex vivo organ culture lost their transparency immediately after alkali burn, and only the groups treated with MSCs and MSCs encapsulated within PEG-collagen hydrogels recovered some transparency after 7 days. Immunohistochemical analysis revealed increased expression of vimentin in the anterior corneal stroma of the group without treatment indicative of fibrotic healing, whereas less stromal vimentin was detected in the group containing MSCs encapsulated within the PEG-collagen hydrogels. Conclusions PEG-collagen hydrogels enable the encapsulation of viable MSCs capable of releasing secreted factors onto the ocular surface. Encapsulating MSCs within PEG-collagen hydrogels may be a promising method for delivering their therapeutic benefits in cases of ocular inflammatory diseases, such as alkali burn injuries.

At high O2 tensions, as encountered prehistorically, Chlamydomonas reinhardtii elevated levels of NPQ-related proteins and LHCSR3 had an important function in protecting PSI., Non-photochemical quenching (NPQ) helps dissipate surplus light energy, preventing formation of reactive oxygen species (ROS). In Chlamydomonas reinhardtii, the thylakoid membrane protein LHCSR3 is involved in pH-dependent (qE-type) NPQ, lacking in the npq4 mutant. Preventing PSII repair revealed that npq4 lost PSII activity faster than the wild type (WT) in elevated O2, while no difference between strains was observed in O2-depleted conditions. Low Fv/Fm values remained 1.5 h after moving cells out of high light, and this qH-type quenching was independent of LHCSR3 and not accompanied by losses of maximum PSII activity. Culturing cells in historic O2 atmospheres (30–35%) increased the qE of cells, due to increased LHCSR1 and PsbS levels, and LHCSR3 in the WT, showing that atmospheric O2 tensions regulate qE capacity. Colony growth of npq4 was severely restricted at elevated O2, and npq4 accumulated more reactive electrophile species (RES) than the WT, which could damage PSI. Levels of PsaA (PSI) were lower in npq4 grown at 35% O2, while PsbA (PSII) levels remained stable. We conclude that even at high O2 concentrations, the PSII repair cycle is sufficient to maintain net levels of PSII. However, LHCSR3 has an important function in protecting PSI against O2-mediated damage, such as via RES.

Seedlings of Pterocarpus indicus were grown in both well-watered and drought stress conditions in phytotron. Seedlings grown under well-watered and drought stress conditions were exposed to either combined or without ozone of 200 ppb for one month. First, the physiological responses to elevated ozone levels indicated a decreased biomass. The seedlings grown in arid soil and exposed to ozone showed less biomass than those grown in arid soil but not exposed to ozone. Moreover, all the seedlings except the well-watered and unexposed ones showed a significantly lower photosynthetic rate (PN) over time. However, with the accumulation of ozone injuries, the antioxidant enzyme activities increased overall. In the study results, when exposed to ozone, the well-watered seedlings exhibited more antioxidative enzyme activity than did the seedlings grown in arid soil. Generally, P. indicus in arid soil suffered less damage from elevated ozone than did the well-watered plants.

PURPOSE: While meibography has proven useful in identifying structural changes in the meibomian gland (MG), little is known regarding the MG spectral transmission and absorption characteristics. The purpose of this study was to measure the transmission/absorption spectra of the MG compared to other eyelid tissues. METHODS: Human and rabbit eyelids were fixed in paraformaldehyde, serial sectioned (50 μm) using a cryotome and imaged by brightfield and reflectance microscopy. Eyelid regions (MG, muscle, tarsus and dermis) were then illuminated by a 100 μm spot using a infrared enhanced white light source. Transmission spectra over a 550–950 nm range were then measured using a spectrometer and differences compared using two-way analysis of variance. RESULTS: Brightfield microscopy of both human and rabbit eyelid tissue showed a marked decrease in light transmission for MG acini compared to other eyelid tissues. In rabbit, the dermis showed 5 × and the muscle showed 2 × more light transmission compared to MG (P < .001 and P < .001, respectively). For human, the muscle showed 14 × and the tarsus showed 84 × more light transmission compared to MG (P < .01 and P < .001, respectively). No specific spectral region of light absorption could be detected in either rabbit or human MG. Loss of light transmission in MG was localized to acini containing small lipid droplets, averaging 2.7 ± 0.8 μm in diameter. CONCLUSIONS: The data suggest that light transmission is dramatically reduced in the acini due to light scattering by small lipid droplets, suggesting that Meibography detects active lipid synthesis in differentiating meibocytes.

The present study was conducted to examine the response of Pterocarpus indicus to elevated ozone by comparing the effects between well-watered and soil drought treatments. The ozone concentration was 200 ppb and the exposure was conducted for a month with well-watered and soil drought treatments in chamber environmental conditions. Elevated ozone-induced visible injuries to P. indicus varied among the well-watered environmental conditions, with the appearance of light green chlorosis in the SWO seedlings over time. However, in the SDO seedlings, no difference was visible between drought and drought with ozone treatment. Shriveled leaves were evident because of the water deficiency. Moreover, elevated ozone induced leaf stomata injury and a harsh abaxial surface, whereas drought stress induced less injured stomata because of stomata closing. In addition, the palisade tissues exhibited smaller chloroplasts, damaged thylakoid, increased plastoglobuli, decreased starch grain and thinner cell walls on the upper leaf surface. Thus, the SDO seedlings were less influenced by elevated ozone stress than SWO seedlings due to the reduced ozone absorption which caused by stomata closing to prevent water loss.

To investigate the role of ATP-sensitive K+(KATP) channels on pacemaker activity in interstitial cells of Cajal (ICC), whole-cell patch clamping, RT-PCR, and intracellular Ca2+([Ca2+]i) imaging were performed in cultured colonic ICC. Pinacidil (a K+ channel opener) hyperpolarized the membrane and inhibited the generation of pacemaker potential, and this effect was reversed by glibenclamide (a KATP channel blocker). RT-PCR showed that Kir 6.1 and SUR2B were expressed in Ano-1 positive colonic ICC. Glibenclamide depolarized the membrane and increased pacemaker potential frequency. However, 5-hydroxydecanoic acid (a mitochondrial KATP channel blocker) had no effects on pacemaker potentials. Phorbol 12-myristate 13-acetate (PMA; a protein kinase C activator) blocked the pinacidil-induced effects, and PMA alone depolarized the membrane and increased pacemaker potential frequency. Cell-permeable 8-bromo-cyclic AMP also increased pacemaker potential frequency. Recordings of spontaneous intracellular Ca2+([Ca2+]i) oscillations showed that glibenclamide increased the frequency of [Ca2+]i oscillations. In small intestinal ICC, glibenclamide alone did not alter the generation of pacemaker potentials, and Kir 6.2 and SUR2B were expressed in Ano-1 positive ICC. Therefore, KATP channels in colonic ICC are activated in resting state and play an important role in maintaining resting membrane potential.

The therapeutic effects of secreted factors (secretome) produced by bone marrow-derived human mesenchymal stem cells (MSCs) were evaluated as a function of their growth in 2D culture conditions and on 3D electrospun fiber scaffolds.Electrospun fiber scaffolds composed of polycaprolactone and gelatin were fabricated to provide a 3D microenvironment for MSCs, and their mechanical properties were optimized to be similar to corneal tissue. The secretome produced by the MSCs cultured on 3D fiber matrices versus 2D culture dishes were analyzed by Luminex immunoassay, and the secretome of MSCs cultured on the 3D versus 2D substrates showed substantial compositional differences. Concentrations of factors such as HGF and ICAM-1 were increased over 5 times in 3D cultures compared to 2D cultures. In vitro proliferation and scratch-based wound healing assays were performed to compare the effects of the secretome on corneal fibroblast cells (CFCs) when delivered synchronously from co-cultured MSCs through a trans-well co-culture system versus asynchronously after harvesting the factors separately and adding them to the media. Cell viability of CFCs was sustained for 6 days when co-cultured with MSCs seeded on the fibers but decreased with time under other conditions. Scratch assays showed 95% closure at 48 hr when CFCs were co-cultured with MSCs seeded on fibers, while the control group only exhibited 50% closure at 48 hr. Electrospun fibers seeded with MSCs were then applied to a wounded rabbit corneal organ culture system, and MSCs seeded on fibers promoted faster epithelialization and less scarring. Corneas were fixed and stained for alpha smooth muscle actin (α-SMA) and keratan sulfate (KS), and then analyzed by confocal microscopy. Immunostaining showed that expression of alpha-SMA was lower in corneas treated with MSCs seeded on fibers, suggesting suppression of myofibroblastic transformation. KS expression was higher in corneas treated with MSCs seeded on fibers, suggesting that keratocytic phenotype was preserved.MSCs cultured on electrospun fibers facilitate wound healing in CFCs and on explanted corneas through differential secretome profiles compared to MSCs cultured on 2D substrates. Future work is merited to further understand the nature and basis of these differences and their effects in animal models.

In this study, an in situ forming corneal stromal substitute based on collagen type I crosslinked by bio-orthogonal strain-promoted azide-alkyne cycloaddition (SPAAC) is presented. The crosslinked collagen gel has greater transparency compared to non-crosslinked collagen gels. The mechanical properties of the gels are controlled by changing functional group ratios and conjugated collagen concentrations. Higher concentrations of conjugated collagen yield enhances mechanical properties, where the storage modulus increases from 42.39 ± 8.95 to 112.03 ± 3.94 Pa after SPAAC crosslinking. Encapsulated corneal keratocytes grow within the SPAAC-crosslinked gels and corneal keratinocytes are supported on top of the gel surfaces. SPAAC-crosslinked gels support more favorable and stable keratinocyte morphology on their surface compared to non-crosslinked gels likely as a result of more optimal substrate stiffness, gel integrity, and resistance to degradation. SPAAC-crosslinked collagen gels with and without encapsulated keratocytes applied to rabbit corneas in an organ culture model after keratectomy exhibit surface epithelialization with multilayered morphology. The novel in situ forming gel is a promising candidate for lamellar and defect reconstruction of corneal stromal tissue.

In order to improve the dispersibility of nanoparticles in polymer matrix, a feasible processing way for grafting high density hydrophobic polymer on the surface of silicon nitride (Si3N4) nanoparticles was carried out by an atom transfer radical polymerization (ATRP) method. The ATRP initiators were first immobilized on the surface of Si3N4 nanoparticles by using 3-aminopropyltriethoxysilane coupling agent and 2-bromoisobutyryl bromide, and then the polymerization of methyl methacrylate was initiated and propagated on the surface of Si3N4 nanoparticles by ATRP reaction. The chemical groups, size, morphology, grafting content, and colloidal thermal stability of PMMA–Si3N4 particles were studied by Fourier transform infrared (FTIR), transmission electron microscopy (TEM) and thermogravimetric analysis (TGA). Results indicate that the grafting of polymer on the surface of Si3N4 nanoparticles is successful and the grafting content of PMMA reaches 60.4%. Moreover, the grafted polymer chains have changed the surface properties of Si3N4 nanoparticles and enhanced their dispersibility in organic media.

The equilibrium moisture contents (EMC) in seeds of four Echinochloa (E. crus-galli var. crus-galli, E. crus-galli var, echinata, E. crus-galli var. praticola, E. esculenta) were measured at 20oC with equilibration over a series of lithium chloride solutions with relative humidities ranging from 0.11 to 0.8 to determine sorption isotherms and safe storage relative humidity. Standard seed sorption isotherm models i.e. modified Henderson, modified Chung-Pfost, modified Halsey, modified Oswin and Guggenheim-Anderson-deBoer (GAB) equations were adopted to evaluate the goodness of fit to sorption isotherms. This study indicated that EMC of seeds was significantly different in four Echinochloa species at various relative humidity. The modified Oswin equations for E. crus-galli var. crou-galli, E. crus-galli var, echinata, E. esculenta and GAB equation for E. crus-galli var. praticola were adequate models for the EMC data. Seeds of four Echinochloa species have monolayer moisture contents when stored at RH < 0.1. These results show that seed moisture isotherm model should be selected according to genetic variation.

The effects of chronic gamma ray irradiation on Brachypodium distachyon were investigated to lignocellulose composition and enzymatic hydrolysis. Four-week-old plants of B. distachyon were exposed to gamma-ray irradiation at doses of 50, 100, 150, 200, 250, and 300 Gy. Gamma-ray irradiation negatively influenced biomass related morphology including plant height and cell size; however, it increased tiller number and internode diameter. SEM image analysis showed that the cell size of 300-Gy gamma irradiated plants was 45 % lower than that of the control. Destroyed regions on the cell wall surface were observed according to the dose of radiation. Cell wall yield, lignin content, and neutral sugar content were decreased in a dose-dependent manner. Glucose yield via enzymatic hydrolysis was positively correlated with radiation dose. In addition, 250-Gy and 300-Gy irradiated plants showed significantly better saccharification efficiency than non-irradiated controls. Initial glucose yields from 250-Gy- and 300-Gy-irradiated plants were 121.5 and 139.4 % of yields from non-irradiated controls, and final yields were 184.7 and 213.7 % of control yields, respectively. Reverse-transcription polymerase chain reaction (RT-PCR) analysis of lignin pathway-related genes showed increase of F5H, COMT, and CAD expression in 250-Gy- and 300-Gy-irradiated Brachypodium plants. Chemical composition analysis further verified that chronic gamma ray irradiation degraded lignocellulose as a pretreatment.

International audience; The production of reactive oxygen species (ROS) is an unavoidable part of photosynthesis. Stress that accompanies high light levels and low CO2 availability putatively includes enhanced ROS production in the so-called Mehler reaction. Such conditions are thought to encourage O2 to become an electron acceptor at photosystem I, producing the ROS superoxide anion radical (O2·-) and hydrogen peroxide (H2 O2 ). In contrast, here it is shown in Chlamydomonas reinhardtii that CO2 depletion under high light levels lowered cellular H2 O2 production, and that elevated CO2 levels increased H2 O2 production. Using various photosynthetic and mitochondrial mutants of C. reinhardtii, the chloroplast was identified as the main source of elevated H2 O2 production under high CO2 availability. High light levels under low CO2 availability induced photoprotective mechanisms called non-photochemical quenching, or NPQ, including state transitions (qT) and high energy state quenching (qE). The qE-deficient mutant npq4 produced more H2 O2 than wild-type cells under high light levels, although less so under high CO2 availability, whereas it demonstrated equal or greater enzymatic H2 O2 -degrading capacity. The qT-deficient mutant stt7-9 produced the same H2 O2 as wild-type cells under high CO2 availability. Physiological levels of H2 O2 were able to hinder qT and the induction of state 2, providing an explanation for why under high light levels and high CO2 availability wild-type cells behaved like stt7-9 cells stuck in state 1.

Photosynthetic organisms have to tolerate rapid changes in light intensity, which is facilitated by non-photochemical quenching (NPQ) and involves modification of energy transfer from light-harvesting complexes (LHC) to the photosystem reaction centres. NPQ includes dissipating excess light energy to heat (qE) and the reversible coupling of LHCII to photosystems (state transitions/qT), which are considered separate NPQ mechanisms. In the model alga Chlamydomonas reinhardtii the LHCSR3 protein has a well characterised role in qE. Here, it is shown in the npq4 mutant, deficient in LHCSR3, that energy coupling to photosystem II (PSII) more akin to qT is also disrupted, but no major differences in LHC phosphorylation or LHC compositions were found in comparison to wild-type cells. The qT of wild-type cells possessed two kinetically distinguishable phases, with LHCSR3 participating in the more rapid (2O2, which accelerated qE induction, revealing a way that may help C. reinhardtii tolerate a sudden increase in light intensity. Overall, a clear mechanistic overlap between qE and qT is shown.

The reaction of 2,4-dihydroxybenzophenone (DHBP) with N-[4-(chlorocarbonyl)phenyl] maleimide provided a novel monomeric ultraviolet (UV)-absorber in the presence of triethylamine. This reactive maleimide was bound onto the backbone of high density polyethylene (HDPE) by melt-grafting in a mini-mixer. The extent of grafting of the monomeric maleimide was quantified by infrared (IR) spectroscopy. To find the optimal grafting conditions, the dependence between reaction temperature, time and the concentration of the monomeric UV-absorber were investigated on the grafting yields. The photo-stabilizing effect of the grafted HDPE was evaluated in a xenonarc weathering apparatus and compared with HDPE mixed with the UV-absorbers using the carbonyl index of the IR spectrum. The grafted HDPE retarded the light initiated oxidation to a greater extent.

A novel monomeric UV-absorber was prepared by the reaction of 2,4-dihydroxybenzophenone with N-4-chlorocarbonylphenylmaleimide in the presence of triethylamine. This reactive maleimide was grafted onto polypropylene (PP) by the thermal melt-processing in a mini-max-moulder. IR spectroscopic method was used for the quantitative determination of the extent of grafting of monomeric maleimide. To find the optimal reaction conditions, the dependence of reaction temperature and time and the concentration of monomeric UV-absorber was investigated on the grafting yields. The photooxidative effect of the grafted PP was evaluated in the weatherometer comparing to the PP mixed with UVabsorbers using the carbonyl index of IR spectrum. The grafted PP showed an excellent anti-photooxidative effect.

We determined the optimal culture and medium conditions for effective production of mycelial mass and exopolysaccharide from a liquid culture of Mycoleptodonoides aitchisonii in an air-lift bioreactor. The mycelial growth and exopolysaccharide production were found to be optimal at a temperature of 25 °C and pH of 6.5. When 60 g/L of lactose was used as a carbon source, the maximum mycelial growth and exopolysaccharide production were obtained. The polypeptone and yeast extract were the most appropriate nitrogen sources for mycelial growth and exopolysaccharide production. In addition, when a mixture of 20 g/L of polypeptone and 5 g/L of yeast extract was used, the exopolysaccharide production increased 50% compared to that of the sole nitrogen source. CaCl2·2H2O (1.0 g/L) was the most effective mineral source. Using the optimal culture and medium conditions, batch cultures with basal and designed medium on mycelial growth and exopolysaccharide production in a 5 L air-lift bioreactor were carried out for 16 days. The mycelial growth and exopolysaccharide production increased with an increase of culture time at 14 days, and the maximum mycelial growth and exopolysaccharide production were 20.3 and 6.2 g/L, respectively, after 14 days of culture. The developed model in an air-lift bioreactor showed good agreement with experimental data. These results indicate that exopolysaccharide production is associated with the mycelial growth of M. aitchisonii in an air-lift bioreactor.

This study was performed to investigate the antioxidant activities of various fractions of Achyranthis radix extract using various solvents in vitro and antioxidant enzyme activities in vivo . Among the various fractions, when n-butanol was used as the extract solvent the DPPH radical scavenge rate was highest at 31.09 μg/ml. The thiobarbituric acid reaction value using n-butanol fraction was 0.33 at 532 nm after 6 days, representing a decrease of about 42% compared to control group. When the administered concentration of A. radix extract was 400 mg/kg/body weight/day, the xanthine oxidase activity in the liver was 19.5 mU/mg protein, representing a decrease of about 46% compared to the control group. The superoxide dismutase activity in the AR-E2 group was 89.4 Umol/(min mg protein), representing a decrease of about 40% compared to the control group. Catalase and glutathione peroxidase were decreased by about 24% and 58%, respectively, compared to the control group. When n-butanol fraction was used, the nitrite scavenging activity in various fractions at pH 1.2 was highest at 90.2%, an increase of about 90% compared to the control group. The results support the antioxidant potential of A. radix extract for various health supplements, pharmaceuticals, and mutraceuticals.

The purposes of this study were to investigate the effects of the bamboo oil on antioxidative activity and nitrite scavenging activity in vitro and in vivo. When the bamboo oil concentration was increased from 10 to 90 μL/mL, the DPPH scavenging rate increased from 15.6 to 98.2%. However, at bamboo oil concentrations above 110 μL/mL, it was not increased. The superoxide anion radical scavenging rate increased from 54.8 to 89.1% when the bamboo oil concentration increased from 110 to 150 μL/mL. When the bamboo oil was incubated for 20 h, the linoleic acid scavenging rate was approximately 91.2%, similar to that of ascorbic acid. The glutathione production using bamboo oil in vivo was 59.0 μM/g of liver, which was about 6.5-fold higher than that of the control. In the case of the activities of glutathione peroxidase and catalase, they were 16.8 U/mg of protein and 295 KU/mg of protein, respectively, approximately 5.6-fold higher and 3.0-fold higher than that of the control. The nitrite scavenging activity was increased from 60.1 to 93.9% at pH of 1.2 when the saturation time was increased from 2 months to 8 months. However, at more than 10 months of saturation, this level was not increased. These results suggest that the bamboo oil of Phyllostachys nigra var. henonis can be used in bioactive and functional materials.

► Seven flavonol glycosides were isolated from Sicyos angulatus. ► All flavonols were identified as 3,7-O-glycosides of quercetin and kaempferol. ► These flavonoids were first time isolated from the genus Sicyos. ► These flavonoids showed differences with other genera of same tribe the Sicyeae. ► Four of the seven flavonoids have various biological activities.

Annexin A1 (ANX-1), a calcium-dependent, phospholipid binding protein, is known to be involved in diverse cellular processes, including regulation of cell growth and differentiation, apoptosis, and inflammation. The mitogen phorbol 12-myristate 13-acetate (PMA) induces expression and phosphorylation of ANX-1. However, the roles of ANX-1 in PMA-induced signal transduction is unknown. Here, we study the cellular localization of ANX-1 in the PMA-induced signal transduction process. We have found that PMA induces the cleavage of ANX-1 in human embryonic kidney (HEK) 293 cells, and that the cleaved form of ANX-1 translocates to the nucleus. The PMA-induced nuclear translocation of ANX-1 was inhibited by the protein kinase C (PKC)δ-specific inhibitor rottlerin, indicating that PKCδ plays a role in nuclear translocation of the cleaved ANX-1. We propose a novel mechanism of PMA-induced translocation of ANX-1 to the nucleus that may participate in the regulation of cell proliferation and differentiation.

Novel beta-hydroxy propenamides as analogues of the active metabolite of leflunomide (A 771726) were synthesized and evaluated for their inhibitory activity on dihydroorotate dehydrogenase (DHODH) in an investigation into their immunosuppressive activity. Compounds 2a, 3a, and 3h were approximately 4-40 times more potent than leflunomide in their activity while they were-less active than A 771726.

Leukotactin-1 (Lkn-1) is a human CC chemokine that binds to both CC chemokine receptor 1 (CCR1) and CCR3. Structurally, Lkn-1 is distinct from other human CC chemokines in that it has long amino acid residues preceding the first cysteine at the NH(2) terminus, and contains two extra cysteines. NH(2)-terminal amino acids of Lkn-1 were deleted serially, and the effects of each deletion were investigated. In CCR1-expressing cells, serial deletion up to 20 amino acids (Delta20) did not change the calcium flux-inducing activity significantly. Deletion of 24 amino acids (Delta24), however, increased the agonistic potency approximately 100-fold. Deletion of 27 or 28 amino acids also increased the agonistic potency to the same level shown by Delta24. Deletion of 29 amino acids, however, abolished the agonistic activity almost completely showing that at least 3 amino acid residues preceding the first cysteine at the NH(2) terminus are essential for the biological activity of Lkn-1. Loss of agonistic activity was due to impaired binding to CCR1. In CCR3-expressing cells, Delta24 was the only form of Lkn-1 mutants that revealed increased agonistic potency. Our results indicate that posttranslational modification is a potential mechanism for the regulation of biological activity of Lkn-1.

Many different stimuli such as bioactive agents and environmental stresses are known to cause the activation of sphingomyelinase (SMase), which hydrolyzes sphingomyelin to generate ceramide as a second messenger playing a key role in differentiation and apoptosis in various cell types. Here we identified multiple forms of the membrane-associated neutral SMase (N-mSMase) activity in bovine brain. They could be classified into two groups according to extracting agents: group T-mSM-ase, extracted with 0.2% Triton X-100, and group S-mSMase, extracted with 0.5 M (NH 4 ) 2 SO 4 . Group T-mSMase was further separated into four forms of T-mSMase: a, β, γ, and δ, which were extensively purified from 40,000-g pellets of bovine brain homogenates by 3,150-, 5,275-, 1,665-, and 2,556-fold over the membrane extracts, respectively, by sequential use of several column chromatographies. On the other hand, S-mSMase was eluted as two active peaks of S-mSMase ∈ and ζ in a phenyl-5PW hydrophobic HPLC column and further purified by 1,119- and 976-fold over 40,000-g pellets of the homogenates, respectively. These highly purified N-mSMase enzyme preparations migrated as several bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and showed many different features in biochemical properties such as pH dependence, Mg 2+ requirements, and effects of detergents. Taken together, our data strongly suggest that mammalian brain N-mSMase may exist as multiple forms different in both its chromatographic profiles and biochemical properties.

Annexins (ANXs) display regulatory functions in diverse cellular processes, including inflammation, immune suppression, and membrane fusion. However, the exact biological functions of ANXs still remain obscure. Inhibition of phospholipase A(2) (PLA(2)) by ANX-I, a 346-amino acid protein, has been observed in studies with various forms of PLA(2). "Substrate depletion" and "specific interaction" have been proposed for the mechanism of PLA(2) inhibition by ANX-I. Previously, we proposed a specific interaction model for inhibition of a 100-kDa porcine spleen cytosolic form of PLA(2) (cPLA(2)) by ANX-I (Kim, K. M., Kim, D. K., Park, Y. M., and Na, D. S. (1994) FEBS Lett. 343, 251-255). Herein, we present an analysis of the inhibition mechanism of cPLA(2) by ANX-I in detail using ANX-I and its deletion mutants. Deletion mutants were produced in Escherichia coli, and inhibition of cPLA(2) activity was determined. The deletion mutant ANX-I-(1-274), containing the N terminus to amino acid 274, exhibited no cPLA(2) inhibitory activity, whereas the deletion mutant ANX-I-(275-346), containing amino acid 275 to the C terminus, retained full activity. The protein-protein interaction between cPLA(2) and ANX-I was examined using the deletion mutants by immunoprecipitation and mammalian two-hybrid methods. Full-length ANX-I and ANX-I-(275-346) interacted with the calcium-dependent lipid-binding domain of cPLA(2). ANX-I-(1-274) did not interact with cPLA(2). Immunoprecipitation of A549 cell lysate with anti-ANX-I antibody resulted in coprecipitation of cPLA(2). These results are consistent with the specific interaction mechanism rather than the substrate depletion model. ANX-I may function as a negative regulator of cPLA(2) in cellular signal transduction.

A facile synthesis of 2-alkyl-substituted chromanones from ortho-acylphenols and terminal alkynoates is described. The method contains two consecutive processes in one-pot reaction through a DABCO-catalyzed condensation reaction and a KOBut-mediated intramolecular cycloaddition to afford the desired products.

Rac is an important regulatory molecule implicated in c-jun N-terminal kinase (JNK) activation in response to stress and cytokines. However, the signaling events that mediate the activation of JNK by Rac are not yet well characterized. To broaden our understanding of downstream mediators that link Rac signals to the JNK pathway, we investigated whether cytosolic phospholipase A(2) (cPLA(2)) is involved in Rac activation of JNK. In this report we demonstrate that either co-transfection with antisense cPLA(2) oligonucleotide or pretreatment with arachidonyltrifluoromethyl ketone (AACOCF3), a potent and specific inhibitor of cPLA(2), inhibits Rac-mediated JNK activation, implying a potential role of cPLA(2) in Rac-signaling to JNK activation. In accordance with this observation, we demonstrate that the addition of exogenous arachidonic acid (AA), a principal product of Rac-activated cPLA(2), or leukotrienes, products of 5-lipoxygenase (5-LO) of AA, caused a specific stimulation of JNK. Together, our findings suggest that cPLA(2) mediates, at least partly, the signaling cascade by which Rac stimulates JNK.