Search

Background: Casbene synthase (CS) is responsible for the first committed step in the biosynthesis of phorbol esters (PE) in the Euphorbiaceae. PE are abundant in the seeds of the biofuel crop Jatropha curcas and its toxicity precludes the use of the protein-rich cake obtained after oil extraction as an animal feed and the toxicity of the fumes derived from burning PE containing biofuel is also a matter of concern. This toxicity is a major hindrance to exploit the potential of this crop as a source of raw material to produce biodiesel. For this reason, the current research on J. curcas is mainly focused on the understanding of the biosynthesis and site of synthesis of PE, as an avenue for the development of genotypes unable to synthesize PE in its seeds. Results: Here, we present targeted proteomics assays (SRM and PRM) to detect and quantify CS in leaves, endosperm, and roots of two J. curcas genotypes with contrasting levels of PE. These assays were based on the use of reference isotopic labeled synthetic peptides (ILSP) predicted from 12 gene models of CS from the J. curcas genome. Conclusion: Our targeted proteomics methods were able to detect and quantify, for the first time, CS gene products and demonstrate the distribution of CS isoforms only in roots from J. curcas genotypes with a high and low concentration of PE. These methods can be expanded to monitor CS, at the protein level, in different tissues and genotypes of J. curcas. [ABSTRACT FROM AUTHOR]

Autosomal dominant mutations in LRRK2 that enhance kinase activity cause Parkinson's disease. LRRK2 phosphorylates a subset of Rab GTPases including Rab8A and Rab10 within its effector binding motif. Here, we explore whether LRRK1, a less studied homolog of LRRK2 that regulates growth factor receptor trafficking and osteoclast biology might also phosphorylate Rab proteins. Using mass spectrometry, we found that in LRRK1 knock-out cells, phosphorylation of Rab7A at Ser72 was most impacted. This residue lies at the equivalent site targeted by LRRK2 on Rab8A and Rab10. Accordingly, recombinant LRRK1 efficiently phosphorylated Rab7A at Ser72, but not Rab8A or Rab10. Employing a novel phospho-specific antibody, we found that phorbol ester stimulation of mouse embryonic fibroblasts markedly enhanced phosphorylation of Rab7A at Ser72 via LRRK1. We identify two LRRK1 mutations (K746G and I1412T), equivalent to the LRRK2 R1441G and I2020T Parkinson's mutations, that enhance LRRK1 mediated phosphorylation of Rab7A. We demonstrate that two regulators of LRRK2 namely Rab29 and VPS35 [D620N], do not influence LRRK1. Widely used LRRK2 inhibitors do not inhibit LRRK1, but we identify a promiscuous inhibitor termed GZD-824 that inhibits both LRRK1 and LRRK2. The PPM1H Rab phosphatase when overexpressed dephosphorylates Rab7A. Finally, the interaction of Rab7A with its effector RILP is not affected by LRRK1 phosphorylation and we observe that maximal stimulation of the TBK1 or PINK1 pathway does not elevate Rab7A phosphorylation. Altogether, these findings reinforce the idea that the LRRK enzymes have evolved as major regulators of Rab biology with distinct substrate specificity. [ABSTRACT FROM AUTHOR]

Jatropha curcas L. belongs to Euphorbiaceae family, and it synthesizes flavonoid and diterpene compounds that have showed antioxidant, anti-inflammatory, anticancer, antiviral, antimicrobial, antifungal and insecticide activity. Seeds of this plant accumulate phorbol esters, which are tigliane type diterpenes, reported as toxic and, depending on its concentration, toxic and non-toxic varieties has been identified. The aim of this work was to characterize the chemical profile of the extracts from seeds, leaves and callus of both varieties (toxic and non-toxic) of Jatropha curcas, to verify the presence of important compounds in dedifferentiated cells and consider the possibility of using these cultures for the massive production of metabolites. Callus induction was obtained using NAA (1.5 mg L−1) and BAP (1.5 mg L−1) after 21 d for both varieties. Thin layer chromatography analysis showed differences in compounds accumulation in callus from non-toxic variety throughout the time of culture, diterpenes showed an increase along the time, in contrast with flavonoids which decreased. Based on the results obtained through microQTOF-QII spectrometer it is suggested a higher accumulation of phorbol esters, derived from 12-deoxy-16-hydroxy-phorbol (m/z 365 [M+H]+), in callus of 38 d than those of 14 d culture, from both varieties. Unlike flavonoids accumulation, the MS chromatograms analysis allowed to suggest lower accumulation of flavonoids as the culture time progresses, in callus from both varieties. The presence of six glycosylated flavonoids is also suggested in leaf and callus extracts derived from both varieties (toxic and non-toxic), including: apigenin 6-C-α-L-arabinopyranosyl-8-C-β-D-xylopyranoside (m/z 535 [M+H]+), apigenin 4′-O-rhamnoside (m/z 417 [M+H]+), vitexin (m/z 433 [M+H]+), vitexin 4′-O-glucoside-2″-O-rhamnoside (m/z 741 [M+H]+), vicenin-2 (m/z 595 [M+H]+), and vicenin-2,6″-O-glucoside (m/z 757 [M+H]+).

Protein kinase signalling, which transduces external messages to mediate cellular growth and metabolism, is frequently deregulated in human disease, and specifically in cancer. As such, there are 77 kinase inhibitors currently approved for the treatment of human disease by the FDA. Due to their historical association as the receptors for the tumourpromoting phorbol esters, PKC isozymes were initially targeted as oncogenes in cancer. However, a meta-analysis of clinical trials with PKC inhibitors in combination with chemotherapy revealed that these treatments were not advantageous, and instead resulted in poorer outcomes and greater adverse effects. More recent studies suggest that instead of inhibiting PKC, therapies should aim to restore PKC function in cancer: cancer-associated PKC mutations are generally loss-of-function and high PKC protein is protective in many cancers, including most notably KRAS-driven cancers. These recent findings have reframed PKC as having a tumour suppressive function. This review focusses on a potential new mechanism of restoring PKC function in cancer -- through targeting of its negative regulator, the Ser/Thr protein phosphatase PHLPP. This phosphatase regulates PKC steady-state levels by regulating the phosphorylation of a key site, the hydrophobic motif, whose phosphorylation is necessary for the stability of the enzyme. We also consider whether the phosphorylation of the potent oncogene KRAS provides a mechanism by which high PKC expression may be protective in KRAS-driven human cancers. [ABSTRACT FROM AUTHOR]

Pitaya (Stenocereus griseus H.) is an exotic fruit produced in some regions of Mexico, where it is mainly consumed fresh and valued in local markets. Its pulp has an intense red color, sweet flavor, small and easily chewable seeds, and compounds with antioxidant capacity. The fruit has thorns that make it difficult to handle and cause mechanical damage. However, if the thorns are removed the shelf-life is decreased and its commercialization in distant markets is more difficult. The objective of this study was to evaluate the effects of 1-methylcyclopropene (1-MCP) and a coating prepared with chitosan (Ch) + oleic acid (OA) were evaluated over the quality of pitayas without thorns in refrigeration (11 ± 1 °C; 93 ± 2 % relative humidity (RH)) for 15 d. The quality parameters studied were respiration pattern, ethylene production, weight loss, titratable acidity, skin firmness, and total soluble solids (TSS). The refrigeration conditions minimized notoriously the weight loss of the fruit. In addition, coating of Ch+OA (CO) maintained the quality parameters and delayed the appearance of fungi, extending the shelflife of without thorns pitayas up to 15 d. However, 1-MCP alone was not capable of preserving the evaluated quality parameters. The best results were observed in (WOT)+CO+1MCP treatment, which delayed the appearance of fungi for 15 d and maintained the quality parameters. [ABSTRACT FROM AUTHOR]

NAD kinase (NADK) is required for the de novo synthesis of NADP+ from NAD+. In neutrophils, NADK plays an essential role by providing sufficient levels of NADPH to support a robust oxidative burst. Activation of NADPH oxidase-2 (NOX-2) in neutrophils by stimulators of protein kinase C (PKC), such as phorbol myristate acetate (PMA), results in the rapid generation of superoxide at the expense of oxidation of NADPH to NADP+. In this study, we measured the levels of pyridine nucleotides following the addition of PMA to neutrophils. PMA elicited a rapid increase in NADP+ in neutrophils, which was not due to oxidation of NADPH, the levels of which also rose. This was mirrored by a rapid reduction in NAD+ levels, suggesting that NADK had been activated. PMA-induced depletion of NAD+ in neutrophils was blocked by PKC inhibitors, but was not dependent on NOX-2, as it was not blocked by the NOX inhibitor, diphenyleneiodonium. PMA also increased NADK activity in neutrophil lysates as well as NADK phosphorylation, as revealed by a monoclonal antibody selective for phospho-NADK. Human recombinant NADK was phosphorylated by PKCδ, resulting in increased immunoreactivity, but unchanged enzyme activity, suggesting that PKC-induced phosphorylation alone is insufficient to increase NADK activity in neutrophils. This leads us to speculate that phosphorylation of NADK promotes the dissociation of an inhibitory molecule from a complex, thereby increasing enzyme activity. Activation of NADK by PKC in phagocytic cells could be critical for the rapid provision of sufficient levels of superoxide for host defence against invading microorganisms. Image 1 • Phorbol myristate acetate increases NADP and NADPH in neutrophils. • The mechanism for this effect is activation of NAD kinase. • NAD kinase in neutrophils is activated by protein kinase C-induced phosphorylation. • Phosphorylation of NAD kinase alone is insufficient to increase enzyme activity. • Phosphorylation might induce the release of an inhibitory protein from NAD kinase. [ABSTRACT FROM AUTHOR]

Thermoplastic elastomers from blends of natural rubber latex NRL and vinylacetate versatic ester copolymer latex VAcVe as a hydrophobic/hydrophilic blend have been prepared by the solution casting technique. The polymer blend was characterized using different characterization techniques such as thermogravimetric analysis, Fourier transform infrared and UV–Vis spectroscopy. The effects of VAcVe blend concentration on the thermal, structural, optical and mechanical properties of the NRL have been studied. The results reveal that the increase in VACVe content led to a more compact structure of the NRL/VAcVe polymer blends resulted in an increase in the disorder in the NRL/VAcVe blends. Further, the effect of gamma radiation on the (50% NRL/50% VAcVe) polymer blend has been studied. Samples from the (50% NRL/50% VAcVe) blend were irradiated with gamma doses of 10 and 30 kGy. It is found that the gamma irradiation leads to the dominance of degradation. In addition, the color changes in the irradiated samples were studied. [ABSTRACT FROM AUTHOR]

We herein report the photoinduced intermolecular umpolung addition of aromatic ketones/aldehydes with α,β-unsaturated esters via ketyl radical intermediates. Following an intramolecular transesterification, a variety of γ-lactone derivatives are readily accessed. Mechanistic investigations demonstrate the significant role of Hantzsch ester, which serves both as the electron and proton donor. [ABSTRACT FROM AUTHOR]

Level of reactive species in blood is an important pathogenic factor in diabetes mellitus leading to dysfunctions of vascular endothelial and smooth muscle cells and coagulation system abnormality. A massive release of reactive species (respiratory burst), catalyzed by NADPH oxidase in blood phagocytes, is not well understood in diabetes. The work aimed to study kinetics of response to microbial particles in blood to specify changes in regulatory mechanisms of generation of reactive species in patients with type 2 diabetes. Production of reactive species in blood and isolated granulocytes was measured by luminol-dependent chemiluminescence. Respiratory burst was initiated by serum opsonized zymosan in blood samples and phorbol ester in cell samples. Kinetic parameters were calculated from experimental kinetic curves of chemiluminescence intensity. ROC curve analysis and mathematical modeling were used to reveal the most significant predictors and clarify specific mechanisms of NADPH oxidase activation. It was shown that kinetic parameters of response to opsonized zymosan (lag-time, response rate, amplitude, production of reactive species) were higher in blood of patients than controls. Amplitude and response rate were the most statistically significant predictors for distinguishing patients and controls at high glucose. It indicated NADPH oxidase activation was the target of hyperglycemia. Mathematical modeling showed hyperglycemia increased stability of NADPH oxidase complex, decreased synchronization of its assembling and elevated neutrophil capacity to phagocytosis in patients. Weak or no dependence of response kinetics on ionomycin concentration was shown in patients indicating changed Ca2+-dependent mechanism of NADPH oxidase activation. Hyperglycemia in type 2 diabetes causes disturbances in mechanisms of NADPH oxidase activation associated with both phagocytosis and the state of intracellular signaling systems, including Ca2+-dependent. We suggest that NADPH oxidase in blood granulocytes can be a promising target for clinical intervention improving management of diabetic complications associated with inflammation. Image 1 • Generation of reactive species was more intensive in blood of patients vs controls. • Amplitude and response rate of respiratory burst were significant predictors. • Modeling showed changes in stability and assembly of NADPH oxidase in diabetes. • Granulocyte capacity to phagocytosis enhanced in patients in model. • Ca2+-signaling in generation of reactive species by granulocytes impaired in T2DM. [ABSTRACT FROM AUTHOR]

Twenty-two diterpenoids (1–22), including two new ones (1, a tigliane-type diterpenoid and 8, an abietane-type diterpenoid) were isolated from the roots of Euphorbia fischeriana Steud. Among them, compounds 4, 7, 12, 14–16, 19–21 are isolated from this plant for the first time. Their structures were elucidated through extensive 1D, 2D NMR and the HRESIMS data. The 13C data of 4 is hereby presented for the first time. The macrocyclic diterpenes 1 and 2 showed marked enhancement of lysosomal biosynthesis after evaluation using lysoTracker staining method. [ABSTRACT FROM AUTHOR]

Jatropha curcas L. belongs to Euphorbiaceae family, and it synthesizes flavonoid and diterpene compounds that have showed antioxidant, anti-inflammatory, anticancer, antiviral, antimicrobial, antifungal and insecticide activity. Seeds of this plant accumulate phorbol esters, which are tigliane type diterpenes, reported as toxic and, depending on its concentration, toxic and non-toxic varieties has been identified. The aim of this work was to characterize the chemical profile of the extracts from seeds, leaves and callus of both varieties (toxic and non-toxic) of Jatropha curcas, to verify the presence of important compounds in dedifferentiated cells and consider the possibility of using these cultures for the massive production of metabolites. Callus induction was obtained using NAA (1.5 mg L-1) and BAP (1.5 mg L-1) after 21 d for both varieties. Thin layer chromatography analysis showed differences in compounds accumulation in callus from non-toxic variety throughout the time of culture, diterpenes showed an increase along the time, in contrast with flavonoids which decreased. Based on the results obtained through microQTOF-QII spectrometer it is suggested a higher accumulation of phorbol esters, derived from 12-deoxy-16-hydroxy-phorbol (m/z 365 [M+H]+), in callus of 38 d than those of 14 d culture, from both varieties. Unlike flavonoids accumulation, the MS chromatograms analysis allowed to suggest lower accumulation of flavonoids as the culture time progresses, in callus from both varieties. The presence of six glycosylated flavonoids is also suggested in leaf and callus extracts derived from both varieties (toxic and non-toxic), including: apigenin 6-C-α-L-arabinopyranosyl-8-C-β-D-xylopyranoside (m/z 535 [M+H]+), apigenin 4'-O-rhamnoside (m/z 417 [M+H]+), vitexin (m/z 433 [M+H]+), vitexin 4/-O-glucoside-2//-O-rhamnoside (m/z 741 [M+H]+), vicenin-2 (m/z 595 [M+H]+), and vicenin-2,6''-O-glucoside (m/z 757 [M+H]+). [ABSTRACT FROM AUTHOR]

The 1,4‐palladium shift strategy allows the functionalization of remote C−H bonds that are difficult to reach directly. Reported here is a domino reaction proceeding by C(sp3)−H activation, 1,4‐palladium shift, and amino‐ or alkoxycarbonylation, which generates a variety of amides and esters bearing a quaternary β‐carbon atom. Mechanistic studies showed that the aminocarbonylation of the σ‐alkylpalladium intermediate arising from the palladium shift is fast using PPh3 as the ligand, and leads to the amide rather than the previously reported indanone product. [ABSTRACT FROM AUTHOR]

The tigliane ring system, which encompasses iconic members such as phorbol and TPA, is widely renowned due to numerous observations of displaying potent biological activity, and subsequent use as mainstream biochemical tools. Traditionally, naturally occurring phorboids are regarded as tumor promotors through PKC activation, although in recent times more highly oxidized natural derivatives have been identified as anti‐tumor agents. In the view that only limited synthetic investigations toward skeletal stereochemical modification have been undertaken, non‐natural systems could be useful for a better understanding of the tigliane pharmacophore via interrogation of cellular sensitivity. In this context the concise construction of a number of highly functionalized non‐natural D‐ring inverted phorbol esters were synthesized, via a rhodium‐catalyzed [4+3] cycloaddition, and biologically evaluated using a range of cancer cell lines. The biological results highlight the notion that subtle changes in structure have dramatic effects on potency. Furthermore, although the non‐natural derivatives did not outcompete the natural systems in the PKC‐activation sensitive MCF7 cancer cell line, they outperformed in other cancer cell lines (MM96L and CAL27). This observation strongly suggested an alternate mode of action not involving activation of PKC, but instead involves thiol addition as indicated by glutathione addition and NF‐κB reporter activity. [ABSTRACT FROM AUTHOR]

• Development of NiO–Fe 2 O 3 and NiO–ZnO supported MWCNT catalysts • Weak + medium acid acidic sites main factor affecting deoxygenation activity • NiO–Fe 2 O 3 /MWCNT catalyst yielded 89 % hydrocarbon and 79 % n -(C 15 +C 17) selectivity • Product selective toward alkanes species. • Nanosize carbon support promote better diesel quality Jatropha curcas is a highly toxic plant that produces seed containing viscous oil with productivity (2 ton/ha), it grows in tropical and sub-tropical regions and offer greater adaptability to a wide range of climatic and soil conditions. Its oils have been noted as an important alternative to produce green diesel via deoxygenation reaction. This study, deoxygenation of jatropha curcas oil (JCO) was carried out over NiO–Fe 2 O 3 and NiO–ZnO catalysts that supported onto multi-walled carbon nanotube (MWCNT). It had found that high Fe and Zn dosages were ineffective in deoxygenation and greatest activity was observed on NiO (20) Fe 2 O 3(5) /MWCNT catalyst. Structure-activity correlations revealed that low metal loading, large density of weak + medium acidic sites and strong basic sites play key role in enhancing the catalytic activities and n -(C 15 +C 17) selectivity. Comparing carbon nanostructures and carbon micron size supported NiO-Fe 2 O 3 revealed that green diesel obtained from NiO–Fe 2 O 3 /MWCNT catalysed deoxygenation had the highest heating value and the lowest amounts of oxygen content. Thereby, it confirmed the importance of carbon nanostructure as the catalyst support in improving the diesel quality. Considering the high reusability of NiO-Fe 2 O 3 /MWCNT (6 consecutive runs) and superior green diesel properties (flash point, cloud properties and cetane index) demonstrated the NiO–Fe 2 O 3 /MWCNT catalyst offers great option in producing excellent properties of green diesel for energy sector. [ABSTRACT FROM AUTHOR]

Extracellular calcium (Ca2+) and store‐operated Ca2+ entry (SOCE) govern homoeostasis in the mammalian epidermis. Multiple microRNAs (miRNA) also regulate epidermal differentiation, and raised external Ca2+ modulates the expression of several such miRNAs in keratinocytes. However, little is known about the regulation of miR‐184 in keratinocytes or the roles of miR‐184 in keratinocyte differentiation. Here we report that exogenous Ca2+ stimulates miR‐184 expression in primary epidermal keratinocytes and that this occurs in a SOCE‐dependent manner. Levels of miR‐184 were raised by about 30‐fold after exposure to 1.5 mM Ca2+ for 5 days. In contrast, neither phorbol ester nor 1,25‐dihydroxyvitamin D3 had any effect on miR‐184 levels. Pharmacologic and genetic inhibitors of SOCE abrogated Ca2+‐dependent miR‐184 induction by 70% or more. Ectopic miR‐184 inhibited keratinocyte proliferation and led to a fourfold increase in the expression of involucrin, a marker of early keratinocyte differentiation. Exogenous miR‐184 also triggered a threefold rise in levels of cyclin E and doubled the levels of γH2AX, a marker of DNA double‐strand breaks. The p21 cyclin‐dependent kinase inhibitor, which supports keratinocyte growth arrest, was also induced by miR‐184. Together our findings point to an SOCE:miR‐184 pathway that targets a cyclin E/DNA damage regulatory node to facilitate keratinocyte differentiation. [ABSTRACT FROM AUTHOR]

Previously, we showed that modulation of the energy barrier for synaptic vesicle fusion boosts release rates supralinearly (Schotten, 2015). Here we show that mouse hippocampal synapses employ this principle to trigger Ca2+-dependent vesicle release and post-tetanic potentiation (PTP). We assess energy barrier changes by fitting release kinetics in response to hypertonic sucrose. Mimicking activation of the C2A domain of the Ca2+-sensor Synaptotagmin-1 (Syt1), by adding a positive charge (Syt1D232N) or increasing its hydrophobicity (Syt14W), lowers the energy barrier. Removing Syt1 or impairing its release inhibitory function (Syt19Pro) increases spontaneous release without affecting the fusion barrier. Both phorbol esters and tetanic stimulation potentiate synaptic strength, and lower the energy barrier equally well in the presence and absence of Syt1. We propose a model where tetanic stimulation activates Syt1-independent mechanisms that lower the energy barrier and act additively with Syt1-dependent mechanisms to produce PTP by exerting multiplicative effects on release rates. [ABSTRACT FROM AUTHOR]

Batch adsorption experiments have been conducted to investigate the removal of methyl orange from aqueous solution by an activated carbon prepared from prickly pear seed cake by phosphoric acid activation. The adsorption process has been described by using kinetic and isotherm models. The kinetic of adsorption was examined by pseudo-first-order, pseudo-second-order, and intraparticle diffusion models. Adsorption isotherm was modeled using Langmuir, Freundlich, Temkin and Dubinin–Radushkevich isotherms. The adsorption process of methyl orange was well explained by the pseudo-second-order model and Freundlich isotherm. Also, pseudo-n-order model has been applied to estimate the order of adsorption kinetic and it was found equal to 2 which confirm the good accuracy of the pseudo-second order. Moreover, Dubinin–Radushkevich isotherm reveals that the adsorption of methyl orange onto activated carbon was a physisorption process in nature. The adsorption capacity of activated carbon was found to be 336.12 mg/g at temperature 20°C and pH ∼ 7. These results demonstrated that the prickly pear seed cake is a suitable precursor for the preparation of appropriate activated carbon for dyes removal from aqueous solution. [ABSTRACT FROM AUTHOR]

Surface modified poly(methyl acrylate) (PMA) grafted chicken feather fiber was applied as a reinforcement for polylactide (PLA)‐based composite film. The PMA grafted feather fiber with different PMA lengths were obtained for surface initiated Cu(0)‐mediated reversible‐deactivation radical polymerization. PLA‐based composite films with different unmodified or PMA‐grafted chicken feather fiber loading were prepared by solvent casting. Grafting PMA onto feather fiber enhanced interfacial bonding. Cracks and voids between matrix and fiber were eliminated. Thermal stability of these films also improved after introducing PMA. Feather fiber with high graft ratio showed worse dispersity in PLA due to its self‐aggregation. The heterogeneous nucleation agent behavior of feather grafted with longer PMA was also suppressed. Tensile tests showed that after modification, tensile strength and modulus was able to increase twice at 20% fiber loading. However, increasing PMA length improved break elongation but strength and modulus would drop slightly. These results indicated that grafting short PMA chain onto feather fiber surface was an effective and optimized method to improve dispersity and mechanical properties especially for higher filler loading. [ABSTRACT FROM AUTHOR]

Excitatory neurotransmission relies on the precise targeting of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors to the neuronal plasma membrane. Activity-dependent ubiquitination of AMPA receptor (AMPAR) subunits sorts internalised receptors to late endosomes for degradation, which ultimately determines the number of AMPARs on neuronal membrane. Our recent study has demonstrated a functional cross-talk between the phosphorylation and ubiquitination of the GluA1 subunit in mammalian central neurons. However, the existence of such a cross modulation for the GluA2 subunit remains unknown. Here, we have shown that bicuculline induced GluA2 ubiquitination on the same lysine residues (Lys-870 and Lys-882) in the C-terminal as those elicited by the AMPA treatment. Interestingly, bicuculline-induced ubiquitination was markedly enhanced by the phospho-mimetic GluA2 S880E mutant. Pharmacological activation of protein kinase C (PKC) by phorbol ester, which mediates the phosphorylation of GluA2 at Ser-880, augmented bicuculline-induced ubiquitination of GluA2 in cultured neurons. This effect was specific for the GluA2 subunit because phorbol ester did not alter the level of GluA1 ubiquitination. However, phorbol ester-induced enhancement of GluA2 ubiquitination did not require Ser-880 phosphorylation. This suggests that pseudo-phosphorylation of Ser-880 is sufficient but is not necessary for the augmentation of bicuculline-induced GluA2 ubiquitination. Collectively, these data provide the first demonstration of subunit-specific modulation of AMPAR ubiquitination by the PKC-dependent signalling pathway in mammalian central neurons. [ABSTRACT FROM AUTHOR]

Natural selection drives local adaptations of species to biotic or abiotic environmental stresses. As a result, adaptive phenotypic divergence can evolve among related species living in different habitats. However, the genetic foundation of this divergence process remains largely unknown. Two closely related alpine grass species, Stipa capillacea and Stipa purpurea , are distributed in different rainfall regions of northern Tibet. Here, we analyzed the drought tolerance of these two closely related Stipa species, and found that S. purpurea was more resistance to drought stress than S. capillacea. To further understand the genetic diversity behind their adaptation to drought environments, a comprehensive gene repertoire was generated using PacBio isoform and Illumina RNA sequencing technologies. Bioinformatics analyses revealed that differential transcripts were mainly enriched in the wax synthetic pathway, and a threonine residue at position 239 of WSD1 was identified as having undergone positive selection in S. purpurea. Using heterologous expression in the Saccharomyces cerevisiae mutant H1246, site-directed mutagenesis studies demonstrated that a positive selection site results in changes to the wax esters profile. This difference may play an important role in S. purpurea in response to drought conditions, indicating that S. purpurea has evolved specific strategies involving its wax biosynthetic pathway as part of its long-term adaptation to the Qinghai–Tibet Plateau. [ABSTRACT FROM AUTHOR]

This study focuses on the mechanical extraction of Jatropha curcas L. oil using a screw press type expeller. The extraction process was carried under different operating conditions to assess the impact on oil yield, oil recovery, extraction capacity, energy intensity, and the required properties as drop-in biofuel or as biodiesel feedstock. The studied variables were the extraction temperature (50 C, 70 °C and 90 C), rotational speed of the screw (40rpm, 50rpm and 60rpm), and nozzle diameter at the end of the press (11mm, 12 mm and 13 mm). The highest oil yield was obtained at the highest temperature, the lowest rotational speed, and using the smallest nozzle diameter. As expected, throughput of the screw press decreased when oil yield increased, nevertheless power consumption barely changed within the evaluated range of operating conditions. Preferred extraction conditions were 90 °C, 40 rpm and cake outlet nozzle diameter of 11 mm. Under these conditions, extraction pressure was 15.35MPa with a specific energy consumption of 0.09kWh/kgSeed- The corresponding oil yield, oil recovery, and extraction capacity were 25.40%, 56.8% and 8.93 kg/h, respectively. By preheating the seeds at 60 °C during 30 min increases oil recovery 15.7% but reduces extraction capacity 16.9%. According to fuel standards, the extracted oil is suitable as drop-in fuel for diesel engines without further refining. Furthermore, it was experimentally demonstrated that the extracted crude oil can be directly transformed into biodiesel using a traditional alkali-based process [ABSTRACT FROM AUTHOR]

Acyclic enediynes are much easier to be synthesized comparing to natural or synthetic cyclic enediynes. However, the conditions for their thermal reactions to produce free radicals through Bergman cyclization (BC) are typically demanding, limiting their applications as antitumor agents. On the other hand, the acyclic compounds with enyne‐allene structures prone to proceed through Myers‐Saito cyclization (MCS) under much milder conditions. Therefore, it is vital to design acyclic enediynes which can permit propargyl‐allenyl rearrangement and then undergo MSC at low temperature for antitumor applications. In this work, we synthesized a series of maleimide‐based enediyne compounds with propargyl ester functionalities. The facile propargyl‐allenyl tautomerization assisted by the maleimide group enabled the formation of enyne‐allene structures and the subsequent occurrence of MSC. The generation of free radicals, which further raised the reactive oxygen species level in physiological environments, was confirmed by electron paramagnetic resonance analysis. The highly reactive radicals showed excellent DNA‐cleaving ability as confirmed with DNA gel electrophoresis. In addition, this kind of acyclic enediynes effectively inhibited the proliferation of HeLa cells with half inhibition concentration down to submicromolar level, comparable or even superior to many clinically used antitumor agents, showing promising application aspect. [ABSTRACT FROM AUTHOR]

As an important gasotransmitter, hydrogen sulfide having multiple biological roles cannot be easily probed in cells. In this study, a light controllable H2S donor, Nap-Sul-ONB, derived from naphthalimide was developed. Under the irradiation of 365 nm light, a readily controlled stimulus, the donor could release COS to form H2S and exhibit turn on fluorescence to indicate the release of payload and its cellular location. Besides, the ROS scavenging ability and cell protective effect of Nap-Sul-ONB against endogenous and exogenous ROS were studied. The results showed that upon 365 nm light irradiation, Nap-Sul-ONB could reduce the cellular ROS level and increase the survival rate of PMA-treated cells. [ABSTRACT FROM AUTHOR]

Total saponins extracted from Dioscorea collettii (TSD), extracts of the Chinese herb Dioscorea, are thought to exhibit therapeutic benefit in gouty arthritis. However, its exact mechanism remains unclear. The current study aimed to elucidate the underlying mechanisms by investigating the effects of TSD on the inflammation induced by monosodium urate (MSU) crystals in THP-1 macrophages. The viability of THP-1 macrophages was examined using the MTT assay and the levels of inflammatory cytokines, including interleukin (IL)-1ß, IL- 18 and tumor necrosis factor (TNF)-a, released by the cells were quantitatively measured using ELISA kits. The results revealed that the protein level of cluster of differentiation 11b increased in THP-1 cells treated with 100 ng/ml phorbol ester, suggesting that monocytic THP-1 cells were successfully differentiated into macrophages. TSD decreased the levels of inflammatory cytokines, including TNF-a, IL- 18 and IL- 1ß, secreted by THP-1 macrophages. As the release of IL-1ß and IL- 18 is dependent on the NLR family pyrin domain containing 3 (NALP3) inflammasome and caspase-1, the current study investigated the effect of TSD on the aforementioned proteins. The results revealed that TSD decreased the protein levels of NALP3 and apoptosis-associated speck-like, which serve important roles in the assembly of the NALP3 inflammasome. Furthermore, NALP3 inflammasome-related proteins were also decreased by TSD in rotenone induced THP-1 macrophages, TSD inhibited the activation of caspase-1 and rotenone-induced NALP3 inflammasome activation in THP-1 macrophages. The results obtained in the current study revealed that TSD attenuated MSU crystal-induced inflammation by inhibiting rotenone-induced activation of the NALP3 inflammasome and caspase-1, suggesting that these two proteins may be novel targets for the treatment of gouty arthritis. [ABSTRACT FROM AUTHOR]

It is hypothesized that the oxidative/nitrosative stress inhibitory effect of a flavanone is governed by its chemical structure. However, the existing cell‐based antioxidant assays primarily focus on single chemical to initiate toxic species production. In this study, a novel cell model using macrophage treated with a combination of PMA and LPS leading to generation of peroxynitrite was proposed to provide a more real physiological condition. Three flavanones (eriodictyol, naringenin, and pinocembrin) with different number of ortho‐dihydroxyl groups on B‐ring were used to provide a more comprehensive evaluation of the role of chemical structure in the new model. Dihydrorhodamine123 assay, protein immunoblotting, immunofluorescence assay, and in silico analysis by molecular docking between the flavanones and IKKβ catalytic kinase domain at the ATP binding site were employed. Results indicated that the generation of peroxynitrite was decreased at 10 µM of flavanones; eriodictyol was the most effective inhibitor. Western blot analysis and confocal fluorescence image also showed that eriodictyol could inhibit iNOS and p47 protein expressions through the inhibition of NF‐kB translocation and performed the maximal inhibition compared to that of the other groups. In addition, the highest CDOCKER energy values of eriodictyol (38.6703 kcal/mol) confirmed that the 3′,4′‐ortho‐dihydroxylation on the B‐ring played a crucial role in binding with IKKβ kinase domain at ATP binding site. Finally, we propose that the ortho‐dihydroxyl groups on B‐ring of flavanone may influence directly the occupation of the ATP binding site of IKKβ kinase domain leading to the abrogation of peroxynitrite formation in the innovative cell model. [ABSTRACT FROM AUTHOR]

Edible oil such has been used as raw material for biodiesel fuel (BDF). However, if the production of biofuel increases, it may compete with food and livestock feed and the price of oil may rise. Therefore, non-edible Jatropha curcas oil containing toxic phorbol esters has attracted attention. Recently a new BDF process which coproduct is not glycerin but triacetin has been developed. For triacetin can also be used as an anti-knock agent for biodiesel fuel, it is not necessary to remove triacetin from biodiesel fuel. The purpose of this study was to investigate the production of biodiesel fuel and reduction of toxic phorbol esters by transesterification of Jatropha curcas oil with methyl acetate. The formation of monoacetin, diacetin and triacetin was also confirmed by the spectrum estimated as the peaks of those substances. The peak of glycerin was not found among the major peaks of the chromatogram. As the molar ratio of methyl acetate to material oil was increased from 5 to 50, the yield of FAME was increased. The maximum FAME yield was 90 %. and the maximum concentration of FAME in the final product was 0.71 g/g. The concentration of phorbol esters in crude Jatropha curcas oil used in this study was 1.7 mg/g. The phorbol ester concentration of degummed oil decreased by a small amount from the crude oil. Phorbol esters decreased after deacidification and the concentration in phorbol esters in deacidified oil was 1.2 mg/g. The concentration in phorbol esters decreased to 0.22 mg/g after mixing step of oil with potassium methoxide. No phorbol esters were detected in any reaction time in the final product after the transesterification reaction in this study. The process used in this study is superior in that the phorbol ester contained in Jatropha curcas oil can be completely decomposed, compared with the conventional method using methanol as a raw material. [ABSTRACT FROM AUTHOR]

Clinical measurement of neopterin has been extensively used as a marker of inflammation but the in vivo mechanism generating neopterin is poorly understood. Neopterin is described as the oxidation product of 7,8-dihydroneopterin, a potent antioxidant generated by monocyte/macrophages in response to interferon-γ. While peroxyl and hydroxyl scavenging generates dihydroxanthopterin, hypochlorite efficiently oxidises 7,8-dihydroneopterin into neopterin, but this reaction alone does not explain the high levels of neopterin seen in clinical data. Here, we examine whether superoxide scavenging by 7,8-dihydroneopterin generates neopterin. U937 cells incubated with oxLDL showed a time dependent increase superoxide and 7,8-dihydroneopterin oxidation to neopterin. Neopterin generation in oxLDL or phorbol ester treated U937 cells or human monocytes was inhibited by apocynin and PEG-SOD. Addition of the myeloperoxidase inhibitor 4-aminobenzoic acid hydrazide (ABAH) had no effect of the superoxide generation or neopterin formation. 7,8-Dihydroneopterin reacted with superoxide/hydroxy radical mixtures generated by X-ray radiolysis to give neopterin. Formation of neopterin by superoxide derived from the xanthine/xanthine oxidase system was inhibited by superoxide dismutase. Neopterin formation was inhibited by apocynin in phorbol ester treated human carotid plaque rings in tissue culture. These results indicate that 7,8-dihydroneopterin scavenges superoxide and is subsequently oxidised into neopterin in cellular and cell-free experimental systems. Image 1 • 7,8-dihydroneopterin is oxidised to neopterin by superoxide generated by xanthine oxidase. • Superoxide generation in response to oxLDL or PMA was associated with an increase in neopterin. • Neopterin formation was reduced by apocynin in PMA treated macrophages and excised human atheroma. • 7,8-Dihydroneopterin scavenges superoxide and is subsequently oxidised into neopterin. • The research suggests a significant amount of the clinically measured neopterin is a product of superoxide scavenging. [ABSTRACT FROM AUTHOR]

The toxic kernel cake of Jatropha curcas (KCakeJ) is an emerging health and environmental concern. Although phorbol esters are widely recognized as the major toxin of KCakeJ, convincing evidence is absent. Here, we show that rather than phorbol esters an isomeric mixture of 11-hydroxy-9E-octadecenoic acid, 12-hydroxy-10E-octadecenoic acid and 12-hydroxy-10Z-octadecenoic acid (hydroxy-octadecenoic acids, molecular formula C18H34O3) is the major toxic component. The toxicities of hydroxy-octadecenoic acids on experimental animals, e.g. acute lethality, causing inflammation, pulmonary hemorrhage and thrombi, allergies, diarrhea and abortion, are consistent with those on human/animals caused by Jatropha seed and/or KCakeJ. The hydroxyl group and the double bond are essential for hydroxy-octadecenoic acids' toxicity. The main pathway of the toxicity mechanism includes down-regulating UCP3 gene expression, promoting ROS production, thus activating CD62P expression (platelet activation) and mast cell degranulation. The identification of the major toxin of KCakeJ lays a foundation for establishing an environmentally friendly Jatropha biofuel industry. [ABSTRACT FROM AUTHOR]

• PKCα is upregulated in Schwann cells of distal nerve stumps as long as 28 days in rat sciatic nerve transection model. • PKCα promotes Schwann cell proliferation and migration in SCs. • PKCα promotes proliferation and migration via ERK pathway. Wallerian degeneration (WD) and axon regeneration generally take place following peripheral nerve injury (PNI). Schwann cells (SCs) and macrophages play major role in WD. SCs, acting as repair cells and primary signal mediators, dedifferentiate and proliferate to remove the debris, form Büngner's bands and secrete trophic factors during these processes. However, the underlying mechanisms remain poorly understood. Here, we found that protein kinase Cα (PKCα), a serine/threonine kinase, expressed in SCs was significantly up-regulated after PNI. Activating PKCα with phorbol 12-myristate 13-acetate (PMA), a phorbol ester binds and activates PKCα) promoted SCs proliferation and migration. While, silence of PKCα by siRNAs inhibited these processes. PD184352, an inhibitor of MEK1, reversed the effect induced by PMA on SCs. Mechanism studies revealed that PKCα functioned through activating the ERK signaling pathway. Furthermore, PKCα also exhibited a neuroprotective role by upregulating the expression of neurotrophic factors in SCs. To sum up, this study offers novel insights for clarifying our understanding of the involvement of PKCα in the mechanism of peripheral nerve degeneration as well as regeneration. [ABSTRACT FROM AUTHOR]

Background and Objective: Peficitinib pharmacokinetics and pharmacodynamics have been characterized mainly in Caucasian subjects. This study investigated the pharmacokinetics, pharmacodynamics, safety, and tolerability of peficitinib in healthy Japanese subjects compared with Caucasian subjects. Methods: In this single-center, randomized, double-blind, placebo-controlled study, a cohort of healthy Japanese (n = 24) and Caucasian (n = 24) men received a single oral dose of peficitinib (20, 60, or 200 mg) or placebo. Another cohort of Japanese men (n = 24) received peficitinib (10, 30, or 100 mg) or placebo twice daily for 7 days. Pharmacokinetic and pharmacodynamic parameters were assessed, and adverse events (AEs) monitored throughout. Results: Dose proportionality of maximum plasma drug concentration (Cmax) and area under the plasma concentration–time curve extrapolated to infinity (AUCinf) was demonstrated for both ethnicities. The geometric mean ratio for dose-normalized Cmax was 45.7–98.8% higher and AUCinf was 33.8–66.4% higher in Japanese versus Caucasian subjects. Mean peak inhibition of STAT5 phosphorylation was higher in Japanese than Caucasian subjects for a given peficitinib dose, but inhibition was comparable across ethnicities for a given plasma peficitinib concentration. In the multiple-dose study, plasma peficitinib concentrations were similar on day 1 and day 7. All AEs were mild, and none resulted in study discontinuation. Conclusions: Peficitinib was well tolerated at doses up to 200 mg daily for 7 days in healthy Japanese subjects. Dose-proportional exposure was demonstrated across the single-dose range of 20–200 mg, with greater peficitinib exposure in Japanese compared with Caucasian subjects. The pharmacokinetic/pharmacodynamic relationships were considered comparable between these populations. ClinicalTrials.gov identifier: NCT01225224. [ABSTRACT FROM AUTHOR]

A novel method has been established for the construction of C–S bonds using redox-active esters with disulfides in the presence of Ru-photoredox catalyst. This method exhibits remarkable functional group tolerance across a wide scope of substrates. Under mild conditions, a structurally diverse array of aryl alkyl sulfides is successfully and efficiently obtained through decarboxylative cross-coupling. [ABSTRACT FROM AUTHOR]

The possibility of deoxofluorination of β-keto esters using SF4 was investigated. The scope and limitation of the reaction were determined. The efficient method for the synthesis of β,β-difluorocarboxylic acids was elaborated based on the reaction. The set of mentioned acids, being the perspective building blocks for medicinal chemistry, were synthesized on multigram scale. The safety of SF4 use was discussed. The described method does not improve upon the safety of using SF4 , but practical recommendations for working with the reagent are proposed. Despite the hazards of using toxic SF4 , a significant increase of efficacy in the synthesis of medicinal-chemistry-relevant building blocks, based on the reaction, in comparison with earlier described approaches is shown. [ABSTRACT FROM AUTHOR]

Objective: Both innate and adaptive immune system play important roles in the onset and progression of inflammatory bowel diseases (IBDs). However, the significance of natural killer (NK) cells for IBDs remains unclear. To understand the biology of colonic lamina propria natural killer (LPNK) cells in IBDs, we characterized LPNK cell metabolism in a murine acute colitis model. Methods: C57BL/6J mice were fed with 3% dextran sulfate sodium to establish the acute colitis model. Colonic LPNK cells were isolated from mice through flow cytometry. The expression of metabolic genes in LPNK cells was analyzed by transcriptome sequencing and quantitative RT-PCR. Glucose uptake, Seahorse assay, and ATP assay were conducted to assess the metabolic status of LPNK cells. Phos-flow assay was performed to evaluate cell signaling pathways in LPNK cells. In vitro stimulation and cytotoxicity assay were conducted to measure the function of LPNK cells. Results: In acute colitis, LPNK cells upregulated the expression of genes related to glycolysis and oxidative phosphorylation (oxphos), and enhanced glucose uptake capability. Intracellular ATP production, glycolysis and oxphos in LPNK cells were also promoted in acute colitis. mTORC1 signaling was essential for the metabolic reprogramming in LPNK cells in acute colitis. Although LPNK cells of diseased mice exhibited equivalent cytokine profile to normal LPNK cells upon stimulation with phorbol ester or IL-2, LPNK cells of diseased mice were more cytotoxic to target cells than normal LPNK cells. Conclusions: LPNK cells undergo metabolic reprogramming which might be a response to upcoming microbial infection in acute colitis. [ABSTRACT FROM AUTHOR]

FDCA esters are highly relevant biobased alternatives for currently used benzene dicarboxylic acid esters. Despite all the developments on 2,5-FDCA applications, to the best of our knowledge thus far no toxicological data were available for 2,5-FDCA esters. In the present study we aimed to fill this gap, by using an in vitro reporter gene assay approach to compare the activity profile of commonly used phthalates to that of their furan-based counterparts. The assay selection was aimed at the detection of endocrine activity, since several phthalates are heavily scrutinised for their endocrine disrupting properties. However, to avoid missing other relevant toxicological endpoints, several assays able to detect various forms of cellular stress were also included in the panel. The results showed that the (ortho)benzene dicarboxylic acid esters were predominantly active on several of the endocrine assays. In comparison, six of the seven furan dicarboxylic acid based diesters tested here showed no activity in any of the 13 assays used. Only the isobutyl derivative DIBF showed moderate estrogenic activity on one assay, compared to much more pronounced activities on four assays for the ortho-phthalate analogue. Overall, the results presented in this paper are a strong indication that 2,5-FDCA based diesters in general are not only technically viable alternatives to phthalates, but also offer significant toxicological benefits, which supports a non-regrettable substitution. [ABSTRACT FROM AUTHOR]

A switch for stability and function: The protein tyrosine phosphatase PTPN22 limits T cell receptor (TCR) signaling, and polymorphisms in the PTPN22 locus are associated with an increased risk of autoimmune diseases. Yang et al. showed that phosphorylation of Ser751 protected PTPN22 from degradation by preventing its ubiquitination. Phosphorylation of this site limited the ability of PTPN22 to inhibit TCR signaling by keeping it away from the plasma membrane (where active TCR is located). Phosphorylation of Ser751 enhanced the interaction of PTPN22 with the kinase CSK, and disruption of this interaction promotes the plasma membrane localization of the phosphatase. These results demonstrate how a single phosphorylation event is a switch for the stability and function of PTPN22, an important brake on TCR signaling. The hematopoietic-specific protein tyrosine phosphatase nonreceptor type 22 (PTPN22) is encoded by a major autoimmunity risk gene. PTPN22 inhibits T cell activation by dephosphorylating substrates involved in proximal T cell receptor (TCR) signaling. Here, we found by mass spectrometry that PTPN22 was phosphorylated at Ser751 by PKCα in Jurkat and primary human T cells activated with phorbol ester/ionomycin or antibodies against CD3/CD28. The phosphorylation of PTPN22 at Ser751 prolonged its half-life by inhibiting K48-linked ubiquitination and impairing recruitment of the phosphatase to the plasma membrane, which is necessary to inhibit proximal TCR signaling. Additionally, the phosphorylation of PTPN22 at Ser751 enhanced the interaction of PTPN22 with the carboxyl-terminal Src kinase (CSK), an interaction that is impaired by the PTPN22 R620W variant associated with autoimmune disease. The phosphorylation of Ser751 did not affect the recruitment of PTPN22 R620W to the plasma membrane but protected this mutant from degradation. Together, out data indicate that phosphorylation at Ser751 mediates a reciprocal regulation of PTPN22 stability versus translocation to TCR signaling complexes by CSK-dependent and CSK-independent mechanisms. [ABSTRACT FROM AUTHOR]

Dalbergia granadillo's human health components were studied using PY-GC-MS, TDS-GC-MS, and GC-MS. The composition of known human health functions was studied by reviewing available literature. The 7-Methyl-Z-tetradecen-1-olacetate has the effect of removing heat and relieving cough in the human body and effectively treat both dry cough and sore throat caused by fire; 1,2-Benzene dicar boxy licacid, bis (2-methyl propyl) ester hasacertainanti-cancer activity, pharmaceutical applications can be used for the synthesis of cancer drugs. [ABSTRACT FROM AUTHOR]

Fatty acid esters of 3-monochloropropane-1,2-diol (3-MCPD) and glycidol are potentially carcinogenic and/or genotoxic processing contaminants that are formed during the process of edible oil refining. Because of their toxicological properties, the presence of these compounds in refined oils and foods containing these oils, particularly infant formula, poses a potential food safety concern. For this reason, recent research efforts have focussed on the development of methods for the analysis of MCPD and glycidyl esters in infant formula in order to estimate levels of exposure. This work presents occurrence data for 3-MCPD and glycidyl esters in 222 infant formulas purchased in the United States between December 2017 and January 2019. The results of this study show a wide range of contaminant concentrations across four different manufacturers, with average bound 3-MCPD concentrations ranging from 0.035 µg g−1 to 0.63 µg g−1 and average bound glycidol concentrations ranging from 0.019 µg g−1 to 0.22 µg g−1. The data suggest that manufacturers B and C source palm oil produced with mitigation measures, leading to reduced amounts of 3-MCPD and glycidyl esters in their infant formulas. Additionally, comparison with a previously published study in our laboratory of the occurrence of 3-MCPD and glycidyl esters in infant formula purchased in the U.S. between 2013 and 2016 revealed that, since 2016, contaminant concentrations have decreased in products produced by manufacturers A, B, and C, while contaminant amounts in formulas from manufacturer D have slightly increased. [ABSTRACT FROM AUTHOR]

In this work, the comb-like poly(2-methyl-2-oxazoline) copolymer, poly(2-aminoethyl methacrylate-random-poly(2-methyl-2-oxazoline) (PMOXA-r-AEMA, PMA) is synthesized, and the CuSO4/H2O2-triggered dopamine/PMA co-deposition process is investigated. Ellipsometry, water contact angle (WCA), and X-ray photoelectron spectroscopy (XPS) are used to characterize the thickness, hydrophilicity, and surface composition of PMA-based coatings. PMA is facilely anchored to substrates within 60 min with the assistance of dopamine/polydopamine triggered by CuSO4/H2O2, and the coating thickness can achieve about 13 nm. Anti-protein adsorption and anti-blood platelet adhesion measurements are also studied to verify their antifouling properties. The adsorption capacity of FITC-BSA can be reduced to 5% of the original surface, and PMA-based coatings present excellent protein-resistant properties (∼95% reduction relative to gold surface) according to surface plasmon resonance (SPR) measurement, and it can resist adhesion of almost all blood platelets. Moreover, it is applied to a capillary inner wall for surface modification. An alkaline protein mixture and egg white proteins are successfully separated, and present excellent stability. The relative standard deviation (RSD) of migration time is lower than 0.71%. The practicability of this promising PMA-based coatings with this preparation strategy can be used for further proteomics analysis. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Animal & Poultry Production is the property of Egyptian National Agricultural Library (ENAL) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Literature has shown that diosgenin, a naturally occurring sapogenin, inducedcytotoxic effects in many cancer models. This study investigated the effect of diosgenin on intracellular Ca2+ concentration ([Ca2+]i) and cytotoxicity in PC3 human prostate cancer cells. Diosgenin (250‐1000 μM) caused [Ca2+]i rises which was reduced by Ca2+ removal. Treatment with thapsigargin eliminated diosgenin‐induced [Ca2+]i increases. In contrast, incubation with diosgeninabolished thapsigargin‐caused [Ca2+]i increases. Suppression of phospholipase C with U73122 eliminated diosgenin‐caused [Ca2+]i increases. Diosgenin evoked Mn2+ influx suggesting that diosgenin induced Ca2+ entry. Diosgenin‐induced Ca2+influx was suppressed by PMA, GF109203X, and nifedipine, econazole, or SKF96365. Diosgenin (250‐600 μM) concentration‐dependently decreased cell viability. However, diosgenin‐induced cytotoxicity was not reversed by chelation of cytosolic Ca2+ with BAPTA/AM. Together, diosgenin evoked [Ca2+]i increases via Ca2+ release and Ca2+ influx, and caused Ca2+‐non‐associated deathin PC3 cells. These findings reveal a newtherapeutic potential of diosgenin for human prostate cancer. [ABSTRACT FROM AUTHOR]

Sterol is synthesized from cholesterol which is from the hydrolysis of stored cholesteryl esters. The process of maintaining cholesterol homeostasis is regulated by SREBP2‐STARD4. Lots of researches demonstrated that male steroidogenesis could be interfered by di‐n‐butyl phthalate (DBP) or monobutyl phthalate (MBP). However, mechanisms of MBP exposure in this process have not been uncovered clearly. The objectiveof this study was to explore roles of SREBP2 and STARD4 in cholesteryl estersynthesis stimulated by MBP in mouse Leydig tumor cells (MLTC‐1). MLTC‐1 exposedto 10‐8, 10‐7, 10‐6, 10‐5 M MBP showed that levels of cholestery ester were increased significantly at 10‐7 M MBP. Besides, cholesteryl ester synthesis stimulated by MBP was down‐regulate when STARD4 or SREBP2 were inhibited. Activity of SREBP2 binding to the promoter of STARD4 was increased after MBP exposure. This study suggests that MBP can increase cholesteryl ester synthesis through SREBP2‐STARD4 signal pathway in MLTC‐1 cells. [ABSTRACT FROM AUTHOR]

Diabetes mellitus can exacerbate renal ischemia-reperfusion (I/R) injury (RI/RI) in diabetic rats. Previous studies have shown that Notch signaling is involved in renal disorders. The aim of the present study was to evaluate the protective effect of the Notch inhibitor γ-secretase N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT) on RI/RI in a streptozocin (STZ)-induced diabetic rat model. STZ-induced diabetic rats were randomly grouped for different treatments. Cisplatin was used to trigger the Notch signaling pathway and the animals were preconditioned with DAPT to block the signaling pathway. Renal function, oxidative stress and inflammatory factors were examined. DAPT-treated diabetic rats demonstrated mitigated renal injury and function, antioxidative activity was significantly improved and HIF-1a was upregulated. Notch inhibitor DAPT is a potential therapeutic target to improve the outcome of RI/RI in STZ-induced diabetic rats in part via the regulation of anti-oxidation and HIF-1a. [ABSTRACT FROM AUTHOR]

Jatropha seed cake is a by-product of biodiesel industry which is rich in proteins and carbohydrates and contains many bioactive compounds. Besides having a good protein quality, the essential amino acid content (except lysine) is also high in Jatropha seed cake, and therefore, Jatropha proteins can be exploited for many technical applications. The major hindrance for its use as a protein supplement is the presence of anti-nutritional factors and toxins, especially the phorbol esters. The detoxification of Jatropha seed cake for the removal of phorbol esters has been shown in our earlier studies. Proteins were extracted from the detoxified Jatropha seed cake and analyzed for amino acid content. Various parameters such as temperature, solubilization pH, precipitation pH and extraction time were studied for their effect on protein content, and the process was optimized using response surface methodology among 5 best samples selected based on protein content and protein yield from dry concentrates. The amount of dry matter, protein content and protein yield of 17.1%, 41.98% and 38.66%, respectively, was obtained under the optimized conditions of 60 °C temperature, solubilization pH 11.0, precipitation pH 4.41 and extraction time of 0.78 h. Amino acid contents revealed that the extracted samples contained a higher concentration of essential amino acids, especially leucine (4.40 ± 0.98 g/100 g). The results indicated that the protein extraction procedure from Jatropha seed cake affected the amino acid content, yet the values were close to FAO/WHO reference protein, and thus, the Jatropha proteins can be explored further for their use as feed supplement. [ABSTRACT FROM AUTHOR]

BACKGROUND: Canola oil processing generates as by‐products cakes rich in proteins, which have high nutritional value. These cakes contain an expressive amount of phenolic compounds which are important as antioxidants, but can interfere in proteins nutritional value. The aim of this work was to identify the phenolic compounds present in canola seed cake and to maximize their extraction by investigating the factors: temperature (20–70 °C), ethanol concentration in water (0–85%) and sample/solvent ratio (1:5–1:20). RESULTS: The maximum phenolic compounds amount was obtained at 65 °C, ethanol concentration of 35% and sample/solvent ratio of 1:20. At this condition, more than 95% of these compounds were extracted from the canola seed cake. It was observed that the solvent could be re‐used for additional extractions, reducing the solvent consumption. A large number of phenolic compounds were identified (24 compounds) and quantified, including sinapic acid derivatives, flavonoid glycosides derivatives and major and minor sinapoyl choline derivatives. CONCLUSION: The phenolic compounds can be easily extracted from the canola seed cake, resulting in a cake with low antinutrients content and, in contrast, an extract rich in antioxidant compounds with possible use as a natural source of antioxidants. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

β1-chimaerin belongs to the chimaerin family of GTPase-activating proteins (GAPs) and is encoded by the CHN2 gene, which also encodes theβ2- andβ3-chimaerin isoforms. All chimaerin isoforms have a C1 domain that binds diacylglycerol as well as tumor-promoting phorbol esters and a catalytic GAP domain that inactivates the small GTPase Rac. Nuclear Rac has emerged as a key regulator of various cell functions, including cell division, and has a pathological role by promoting tumorigenesis and metastasis. However, how nuclear Rac is regulated has not been fully addressed. Here, using several approaches, including siRNA-mediated gene silencing, confocal microscopy, and subcellular fractionation, we identified a nuclear variant of β1-chimaerin, β1-Δ7p-chimaerin, that participates in the regulation of nuclear Rac1. We show that β1-Δ7p-chimaerin is a truncated variant generated by alternative splicing at a cryptic splice site in exon 7. We found that, unlike other chimaerin isoforms, β1-Δ7p-chimaerin lacks a functional C1 domain and is not regulated by diacylglycerol. We found that β1-Δ7pchimaerin localizes to the nucleus via a nuclear localization signal in its N terminus. We also identified a key nuclear export signal in β1-chimaerin that is absent in β1-Δ7p-chimaerin, causing nuclear retention of this truncated variant. Functionally analyses revealed that β1-Δ7p-chimaerin inactivates nuclear Rac and negatively regulates the cell cycle. Our results provide important insights into the diversity of chimaerin Rac-GAP regulation and function and highlight a potential mechanism of nuclear Rac inactivation that may play significant roles in pathologies such as cancer. [ABSTRACT FROM AUTHOR]

Upon injury, psychotropic psilocybin‐producing mushrooms instantly develop an intense blue color, the chemical basis and mode of formation of which has remained elusive. We report two enzymes from Psilocybe cubensis that carry out a two‐step cascade to prepare psilocybin for oxidative oligomerization that leads to blue products. The phosphatase PsiP removes the 4‐O‐phosphate group to yield psilocin, while PsiL oxidizes its 4‐hydroxy group. The PsiL reaction was monitored by in situ 13C NMR spectroscopy, which indicated that oxidative coupling of psilocyl residues occurs primarily via C‐5. MS and IR spectroscopy indicated the formation of a heterogeneous mixture of preferentially psilocyl 3‐ to 13‐mers and suggest multiple oligomerization routes, depending on oxidative power and substrate concentration. The results also imply that phosphate ester of psilocybin serves a reversible protective function. [ABSTRACT FROM AUTHOR]

The aim of the present study was to explore the possible role of microRNA-144-5p (miR-144-5p) in rheumatoid arthritis (RA) and the associated mechanism. Following the induction of THP-1 cell differentiation into macrophages by phorbol ester (100 ng/ml) treatment, an in vitro inflammatory model of RA was established by treating the THP-1 macrophages with 1 µg/ml lipopolysaccharide (LPS). The level of miR-144-5p was subsequently measured using reverse transcription-quantitative PCR, which found that the expression of miR-144-5p was significantly reduced in LPS-treated THP-1 macrophages. Bioinformatics analysis and a dual-luciferase reporter assay were used to predict and confirm TLR2 as a direct target of miR-144-5p, respectively. Toll-like receptor 2 (TLR2) was then validated as a target gene of miR-144-5p. The effects of miR-144-5p upregulation and TLR2 silencing on LPS-treated THP-1 macrophages were then determined by transfection with miR-144-5p mimic and TLR2-siRNA, respectively. Cell viability was subsequently measured using a Cell Counting Kit-8 assay, whilst the expression of tumor necrosis factor-α (TNF-α), interleukin (IL)-6 and IL-8 secreted by THP-1 macrophages was measured using ELISA. Western blotting was performed to measure p65 phosphorylation (p-p65) in the NK-κB signaling pathway. It was found that miR-144-5p overexpression reduced macrophage cell viability, reduced the expression of TNF-α, IL-6 and IL-8, and reduced the expression of TLR2 and p-p65 compared with the control group. Likewise, TLR2 silencing also reduced macrophage cell viability and reduced the expression of TNF-α, IL-6 and IL-8 in THP-1 macrophages. In conclusion, the data from the present study suggested that miR-144-5p overexpression reduced THP-1 macrophage cell viability and inhibited the expression of TNF-α, IL-6 and IL-8 in cells, possibly by inhibiting the expression of TLR2 and suppressing the activation of NK-κB signaling. Therefore, miR-144-5p may serve as a novel therapeutic target for the treatment of RA. [ABSTRACT FROM AUTHOR]

Fifteen jatropha lines were obtained from complex crossing between progenies derived from interspecific hybridization of Jatropha curcas and J. integerrima. They were evaluated for variability and association in yield, yield components, oil content, fatty acid composition, phorbol esters content, and growth characters. The major fatty acids in seed were oleic and linoleic acids, with a negative correlation among them. Seed yield showed a positive correlation with number of fruits per inflorescence and seeds per fruit, 100 seed weight and canopy size. A path coefficient analysis showed that number of fruits per inflorescence and canopy width had high positive direct relationship with seed yield per plant. Most characters showed high broad-sense heritability. Clustering by traits classified the breeding lines into five groups with a large distance between groups. The members in each group comprised lines of the same or similar pedigrees. Cluster I was the high seed yield group comprising two accessions (KUJL23 and KUJL18). Clustering based on principle components classified them into four groups. The lines in cluster II and III from cluster analysis were included in one group, while members of the other groups were the same in both clustering methods. [ABSTRACT FROM AUTHOR]

A palladium catalyst under visible light irradiation activates cyclobutanone oxime ester through single electron transfer to induce radical ring opening to generate hybrid cyanoalkyl Pd(I) radical species. Hybrid cyanoalkyl Pd(I) radical species can undergo either β-H elimination to deliver (E)-4-arylbut-3-enenitrile or undergo radical addition with silyl enol ether and enamide to generate δ-cyano ketones. A dual ligand system composed of two phosphine ligands is essential for the high reactivity. [ABSTRACT FROM AUTHOR]