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Agrocybe cylindracea is one of the common sources of functional polysaccharides. But the structure and function of Agrocybe cylindracea polysaccharides obtained by different preparation methods are very different. A comprehensive and detailed study of its fine structure and systematic activity is helpful for its development and utilization. In this study, a cold water-souble polysaccharide (ACP) and an alcohol-soluble polysaccharide (ACAP) from Agrocybe cylindracea were extracted. Their physicochemical property, structural feature and antitumor activity in vitro were compared and investigated. The results showed that ACP (2840.9 kDa) was mainly composed of 1,6-linked-Glcp, 1,3,6-linked-Glcp, 1,6-linked-Galp and T-Glcp and inhibited MGC-803 cells proliferation via inducing tumor cells apoptosis by the cytochrome c/caspases pathway. However, ACAP (1823.9 Da) just contained glucose. By comparison, ACAP exhibited no inhibitory effect on MGC-803 cells in vitro. This work would provide an additional explanation for the structure-activity relationship of polysaccharides from Agrocybe cylindracea and provide a theoretical support for its application in biomedicine industries. [ABSTRACT FROM AUTHOR]

Gleditsia sinensis Lam. , a natural product derived from medicine and food, exhibits outstanding biological activity, with its primary active component being triterpenoid saponin. In this study, a new macromolecular Gleditsia Saponin (GS) with a molecular weight of 27.562 kDa was extracted from Gleditsia sinensis Lam.. The study focused on examining the impact of GS on the suppression of cell growth and the initiation of apoptosis in leukemia K562 and L1210 cells. The results showed that GS notably suppressed the growth of leukemia cells in a manner reliant on concentration. Morphological and Annexin V-FITC/PI staining analyses revealed characteristic features of apoptosis in leukemia cells treated with GS. Moreover, cell cycle analysis indicated that GS effectively arrested leukemia cell progression at the G0/G1 phase. Mitochondrial membrane potential (ΔΨm) was disrupted, and abnormal accumulation of reactive oxygen species was also detected. In addition, GS could induce cell apoptosis through the Fas-mediated death receptor pathway and mitochondrial pathway, leading to the blocking of leukemia cells in the G1 phase. In summary, these results provide theoretical backing and fresh perspectives for the development and utilization of GS as a functional health food. • A new macromolecular Gleditsia Saponin (GS) with molecular weight of 27.562 kDa was obtained. • The purity and characteristic absorption peaks of GS were comfirmed by HPGPC, MALDI-TOF-MS and FT-IR. • GS can induce apoptosis of leukemia cells through Fas-mediated death receptor pathway and mitochondrial pathway. • GS, more economical and common than ginsenosides, offers new ideas for health food. [ABSTRACT FROM AUTHOR]

In this study, a novel low molecular weight polysaccharide (named LMW-BSP) was extracted from Bletilla striata at 4 °C. The results of structural characteristics analysis showed that LMW-BSP was a 23 kDa neutral polysaccharide contained glucose and mannose at a molar ratio of 1.00:1.26. Structural investigations of the periodate oxidation studies, Smith-degradation as well as methylation were performed, and combined with 1D and 2D NMR spectroscopy, the main chain residues sequence of LMW-BSP was concluded to be: α-D-Man p -(1 → 3)-β-D-Man p -(1 → [4)-β-D-Glc p -(1] 2 → 4)-β-D-Man p -(1 → 3)-β-D-Man p -(1→. Moreover, the antitumor activity of LMW-BSP was evaluated in H22 tumor-bearing mice. And the results suggested that LMW-BSP could effectively improve immune cells activities and lymphocytes subsets proportions dose-dependently in tumor-bearing mice, leading to the apoptosis of H22 cells via G1 phase arrested. LMW-BSP inhibited tumor growth and exhibited antitumor effects in vivo. And it supported considering the novel polysaccharide as a potential drug component in hepatocellular carcinoma treatment. [ABSTRACT FROM AUTHOR]

Purpose: Recent studies reported therapeutic effects of Smad7 on oral mucositis in mice without compromising radiation therapy-induced cancer cell killing in neighboring oral cancer. This study aims to assess whether a Smad7-based biologic can treat oral mucositis in a clinically relevant setting by establishing an oral mucositis model in dogs and analyzing molecular targets.Methods and Materials: We created a truncated human Smad7 protein fused with the cell-penetrating Tat tag (Tat-PYC-Smad7). We used intensity modulated radiation therapy to induce oral mucositis in dogs and applied Tat-PYC-Smad7 to the oral mucosa in dose-finding studies after intensity modulated radiation therapy. Clinical outcomes were evaluated. Molecular targets were analyzed in biopsies and serum samples.Results: Tat-PYC-Smad7 treatment significantly shortened the duration of grade 3 oral mucositis based on double-blinded Veterinary Radiation Therapy Oncology Group scores and histopathology evaluations. Topically applied Tat-PYC-Smad7 primarily penetrated epithelial cells and was undetectable in serum. NanoString nCounter Canine IO Panel identified that, compared to the vehicle samples, top molecular changes in Tat-PYC-Smad7 treated samples include reductions in inflammation and cell death and increases in cell growth and DNA repair. Consistently, immunostaining shows that Tat-PYC-Smad7 reduced DNA damage and neutrophil infiltration with attenuated TGF-β and NFκB signaling. Furthermore, IL-1β and TNF-α were lower in Tat-PYC-Smad7 treated mucosa and serum samples compared to those in vehicle controls.Conclusions: Topical Tat-PYC-Smad7 application demonstrated therapeutic effects on oral mucositis induced by intensity modulated radiation therapy in dogs. The local effects of Tat-PYC-Smad7 targeted molecules involved in oral mucositis pathogenesis as well as reduced systemic inflammatory cytokines. [ABSTRACT FROM AUTHOR]

The purposes of this article were to investigate the influences of synthetic strategy on the CuO crystals using cellulose as substrate. Exploring the influences of different synthesis methods is of great importance for the understanding of the growth mechanism of CuO crystals. In this article, the products were characterized by means of X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), differential thermal analysis (DTA), and cell cytotoxicity. Experimental results demonstrated the obvious differences in the CuO crystals between microwave-assisted hydrothermal and hydrothermal methods in the existence of cellulose. These researches do a favor for the understanding of growth mechanism of CuO crystals with different synthesis methods. [ABSTRACT FROM AUTHOR]

Abstract: Combined with the Measure While Drilling (MWD) system of the new kilometers directional drilling rig and the method of directional drilling technology, this paper uses animated video technology of product to simulate the major workflow and the actions of whole technology and equipment which used to construct drilling hole for gas drainage in the coal mine, and the production of the animated videos make the related personnel establish the full visual impression in the practical application course of whole directional drilling technology and equipment, and it also could be used for the propaganda of related technology products and personnel technical training, and help to promote the application of this technology. [Copyright &y& Elsevier]

Nanog regulates human and mouse embryonic stem (ES) cell self-renewal activity. Activation of ERKs signaling negatively regulates ES cell self-renewal and induces differentiation, but the mechanisms are not understood. We found that ERK1 binds and phosphorylates Nanog. Activation of MEK/ERKs signaling and phosphorylation of Nanog inhibit Nanog transactivation, inducing ES cell differentiation. Conversely, suppression of MEK/ERKs signaling enhances Nanog transactivation to inhibit ES cell differentiation. We observed that phosphorylation of Nanog by ERK1 decreases Nanog stability through ubiquitination-mediated protein degradation. Further, we found that this phosphorylation induces binding of FBXW8 with Nanog to reduce Nanog protein stability. Overall, our results demonstrated that ERKs-mediated Nanog phosphorylation plays an important role in self-renewal of ES cells through FBXW8-mediated Nanog protein stability. [ABSTRACT FROM AUTHOR]

Lipofuscin accumulation in retinal pigment epithelial (RPE) cells of the eye implicates the etiologies of Stargardt disease and age-related macular degeneration, a leading cause of blindness in the elderly. Here, we have identified a previously unknown RPE lipofuscin component. By one- and two-dimensional NMR techniques and mass spectrometry, we confirmed that this compound is a new type of pyridinium bisretinoid presenting an unusual structure, in which two polyenic side chains are attached to adjacent carbons of a pyridinium ring. This pigment is a light-induced isomer of iso A2E, rather than A2E, referred to as iisoA2E. This pigment is a fluorescent lipofuscin compound with absorbance maxima at ~430 and 352 nm detected in human, pig, mouse, and bovine eyes. Formation of iisoA2E was found in reaction mixtures of all-trans-retinal and ethanolamine. Excess intracellular accumulation of this adduct in RPE cells in vitro leads to a significant loss of cell viability and caused membrane damage. Phospholipase D-mediated phosphodiester cleavage of the A2PE series generated isoA2E and iisoA2E, in addition to A2E, thus corroborating the presence of isoA2PE and iisoA2PE that may serve as biosynthetic precursors of isoA2E and iisoA2E. [ABSTRACT FROM AUTHOR]

Abstract: Introduction: Inclusion-body myopathy (IBM) with Paget''s disease of bone (PDB) and frontotemporal dementia (FTD), designated as IBMPFD, is a rare, autosomal dominant disorder (MIM 605382). IBMPFD is caused by mutations in the gene that encode valosin-containing protein (VCP). We investigated a Chinese family in which multiple members were diagnosed with PDB and suffered from weakness of the limbs. However, no members of this family were diagnosed with FTD. We made a preliminary diagnosis of PDB, but failed to identify an SQSTM1 mutation in any of the patients. We used whole-exome sequencing to identify the pathogenic gene mutation affecting the Chinese male proband. Materials and methods: Altogether, 254 subjects, including one 56-year-old male proband, four affected, related individuals and additional nine family members from a non-consanguineous Chinese family, and 240 healthy donors were recruited and genomic DNA was extracted. All eight exons and the exon–intron boundaries of the SQSTM1 gene were amplified by polymerase chain reaction (PCR) and directly sequenced in five patients (II13, II4, II5, II8, II9). Using whole-exome sequencing, we identified a novel mutation in VCP as the disease-causing mutation. We confirmed the result by sequencing a 500-bp region of the promoter and the coding region of VCP in all 254 of the participants using Sanger sequencing. Results: No mutation in the SQSTM1 gene was identified in the five patients examined using direct Sanger sequencing. However, through whole-exome sequencing we were able to identify a novel missense mutation in exon 3 of the VCP gene (p.Gly97Glu) in the Chinese male proband. This mutation was confirmed using Sanger sequencing. The proband, four affected individuals and three unaffected individuals carried this mutation. We were able to correctly diagnose the patients with atypical IBMPFD. Structural analysis of the p.Gly97Glu mutation in the VCP protein showed that the affected amino‐acid is located in the interface of the protein. This abnormality may therefore interfere with protein function. Conclusions: This is the first report of a family from China with IBMPFD. A novel VCP mutation was found as the cause of atypical IBMPFD in a Chinese family. Our findings confirm that VCP gene mutations can be a pathogenic cause of IBMPFD. [Copyright &y& Elsevier]

Abstract: Previous studies have suggested that changes in hip geometry increase the risk of hip fracture. The aim of this study was to identify whether body composition were associated with hip geometry or bone mineral density (BMD) in a large sample of Chinese people. A total of 2072 subjects aged 20–79yr (including 700 males and 1372 females) were selected. The following measurements were taken: lumbar spine (L1–4); proximal femur BMD; lean mass (LM); fat mass (FM); and hip geometric parameters, including hip axis length (HAL), cross-sectional moment of inertia (CSMI), cross-sectional area (CSA), neck-shaft angle, and femur strength index (SI) by dual-energy X-ray absorptiometry. FM and LM were positively correlated with HAL, CSMI, and CSA, and negatively correlated with SI in both men and women. Multiple regression analysis showed that leg LM contributions to HAL, CSMI, and CSA variance were 12.6–37.6%. Compared with FM, LM was generally more strongly related to hip geometry and BMD in young and old men and women. Body composition was a good predictor for hip geometry parameter variation and BMD variation. [Copyright &y& Elsevier]

Background and Aims: Hypophosphatasia is a genetic disorder characterized by defective bone and tooth mineralization and a deficiency of serum and bone alkaline phosphatase activity. To date, few studies have identified gene mutations in Chinese patients with hypophosphatasia. We sought to characterize the clinical manifestations and identify the mutations associated with the disease in Chinese hypophosphatasia patients. Methods: All 12 exons and the exon-intron boundaries of the ALPL gene were amplified and directly sequenced in two probands from unrelated Chinese families. The mutation sites were identified in other unaffected members of these two families and 100 healthy controls. Results: In family 1, the proband displayed one novel splice site mutation, c.298-1G>A, which consisted of a homozygous G>A transition at nucleotide 298-1 in intron 4. The proband’s mother displayed the heterozygous G/A ALPL gene mutation, but her father was identified as G/G homozygous. A paternity test ruled out false paternity and therefore confirmed that this splicing mutation occurred de novo either in the paternal germline or in the early development of the patient. In family 2, the proband revealed a novel missense mutation (c.1271T>C) in exon 11, which resulted in p.Val424Ala in the mature ALPL polypeptide. Furthermore, c.298-1G>A and c.1271T>C mutations were not found in unaffected family members of these two Chinese families and 100 unrelated controls. Conclusions: Our study shows that the novel de novo splicing mutation c.298-1G>A in intron 4 and the missense mutation c.1271T>C in exon 11 of the ALPL gene are responsible for hypophosphatasia in some Chinese patients. [Copyright &y& Elsevier]

Cynanchum paniculatum has been applied for various diseases treatments, however, the polysaccharide components especially extracted using ethanol were rarely researched. In this study, ethanol solution with volume fraction of 75% was used for C. paniculatum polysaccharide preparation, and the extraction process optimization, structural characteristics, and bioactivities analysis were investigated. Results showed that extraction yield reached to 4.79 ± 0.24% under extraction temperature of 60 °C, extraction time of 4.5 h, liquid/solid ratio of 30 mL/g. Chemical constitutions and monosaccharide compositions, molecular weight determination results showed that the polysaccharide was mainly composed of glucose and xylose with a molar ratio of 1.00:0.63 (carbohydrate content of 94.91%, molecular weight of 5.1 kDa). Periodate oxidation, smith degradation, methylation, fourier transform infrared spectra and nuclear magnetic resonance analysis demonstrated that the glycosidic bonds configurations were mainly comprised of (1→2)-α- D -Glc p (39.86%), (1→2,4)-α- D -Glc p (10.32%) and (1→4)-β- D -Xyl p (29.58%). The bioactivities determination experiments results suggested that the polysaccharide exhibited strong antioxidant activities and antitumor effects both in vitro and in vivo. These data indicated that the ethanol-extracted polysaccharide from C. paniculatum holds great application prospect as a novel bioactive adjuvant in the future due to the feasibility of industrial production. • The polysaccharide was extracted from Cynanchum paniculatum using 75% ethanol. • The maximum extraction yield was 4.79 ± 0.24% optimized by Box-Behnken design. • The polysaccharide was composed of glucose and xylose (molar ratio of 1.00:0.63). • The polysaccharide was comprised of (1→2)-Glc p (39.86%) and (1→4)-Xyl p (29.58%). • The polysaccharide exhibited strong antioxidant and antitumor effects. [ABSTRACT FROM AUTHOR]

RSK2, an ERK downstream kinase, is a novel mediator of skeletal muscle cell differentiation through its regulation of NFAT3 activity. We found that the N-terminal (amino acids (aa) 1-68) and C-terminal (aa 416-674) kinase domains of RSK2 directly interacted with nuclear localization signal 1, the Ser/Pro repeat, and the polyproline domains (aa 261-365) of NFAT3. Upon A23187 stimulation, RSK2 induced nuclear localization of NFAT3. RSK2 phosphorylated NFAT3 in vitro (Km = 3.559 μM), and activation of NFAT3 by RSK2 enhanced the promoter activity of NFAT3 downstream target genes in vivo. Furthermore, nuclear accumulation of NFAT3 was attenuated markedly in RSK2-/- cells compared with wild-type RSK2+/+ cells. Notably, RSK2 and NFAT3 induced a significant differentiation of C2C12 myoblasts to multinucleated myotubes. Multinucleated myotube differentiation was inhibited by small interfering RNA against RSK2, ERK1/2, or NFAT3. These results demonstrate that RSK2 is an important kinase for NFAT3 in mediating myotube differentiation. [ABSTRACT FROM AUTHOR]

[Display omitted] • Oxidation status of LOs are monitor via phenyl-hexyl based LC column. • Number of Trp and Met residue is critical for oxidative stability of Los. • L-AP is effective in suppressing oxidation of both Trp- and non-Trp containing LOs. Linusorbs (LOs, cyclolinopeptides) are a class of naturally occurring cyclic hydrophobic peptides found in flaxseed oil, whose oxidation states indicate the oxidative stability and bitterness of flaxseed oil. Subjected to 63 °C accelerated oxidation, most Met-containing LOs in cold-pressed flaxseed oil entirely depleted by the 6th day except CLP, and MetO 2 -containing LOs became the dominant ones. However, no MetO 2 form of Trp-containing LOs, such as CLD, CLF and CLG, were detected. Given their oxidative kinetics, methionine sulfoxide (MetO) residue in some LOs was less sensitive toward oxidation in the presence of Trp (Tryptophan) group, and the oxidative stability of Met-containing LOs was CLP < CLB < CLL ≈ CLM < CLO, as compared to MetO-containing LOs: CLD < CLE < CLC < CLF ≈ CLG. When antioxidant was added into cold-pressed flaxseed oil to assess the additives' antioxidant effect, no significant difference was observed on oil oxidative indices in early stage except α-tocopherol, where they vary dramatically in suppressing Met oxidation of LOs: L-AP (L-ascorbyl palmitate) > TBHQ (tert -butyl hydroquinone) > γ-tocopherol > carnosic acid > α-tocopherol. Besides its ability to suppress oxidation of Trp-containing LOs, L-AP also exhibits superior antioxidant effect on non-Trp-containing LOs due to its amphiphilic property. Due to the prooxidative effects of both α- and γ-tocopherol on LOs that contain Trp, it has been suggested that tocopherols may repair Trp residue on LOs, leading to increased tendency of MetO residues to oxidize. The findings of this research are critical for elucidating the antioxidative mechanism of LOs, which can further lead to the establishment of strategies in suppressing bitter after taste to produce high-quality flaxseed oil. [ABSTRACT FROM AUTHOR]

Major objectives in research for food and environmental samples include improving robustness, sensitivity, selectivity, and extraction efficiency for polar organic compounds, avoiding matrix effects, and developing faster, simpler, more environment-friendly sample-preparation procedures. In this article, we review novel configurations of solid-phase microextraction (SPME), which make the whole process of analysis more selective, more sensitive and more environment friendly. These techniques include hollow-fiber SPME, magnetic dispersive SPME, and ionic liquid-based SPME. Finally, we look ahead to future potential developments of these configurations of SPME. [ABSTRACT FROM AUTHOR]

In this article, the manganese-containing cellulose nanocomposites were obtained using microcrystalline cellulose and Mn(CH3COO)2·4H2O in the NaOH/urea aqueous solutions by a efficient microwave-assisted method. The effects of the heating time and Mn(CH3COO)2·4H2O concentration on the cellulose nanocomposites were investigated. It was found that the microcrystalline cellulose pretreated with NaOH/urea aqueous solutions played an important role in the phase, shape, and thermal stability of manganese-containing cellulose nanocomposites. Well-crystalline phases of manganese oxides were not observed in the manganese-containing cellulose nanocomposites. Furthermore, well-crystalline phases of manganese oxides were not also observed by thermal treatment of the manganese-containing cellulose nanocomposites at 600 °C for 3 h. These results could be attributed to the restrain effect of cellulose treated with NaOH/urea aqueous solutions. It was supposed the possible mechanism during the phase transformation of cellulose nanocomposites. [ABSTRACT FROM AUTHOR]

Highlights: [•] MWCNTs/SiO2 hollow fibre was fabricated through layer by layer self-assembly. [•] Rapid analysis method of PAEs in food matrix without sample clean-up was established. [•] MWCNTs/SiO2 HF-SPME was applied to extract PAEs in beverage and alcoholic samples. [•] HF-SPME procedure was confirmed by solvent extraction of Chinese national standard. [•] The proposed method has the advantage of simplicity, good accuracy and precision. [Copyright &y& Elsevier]

RSK2 is a widely expressed serine/threonine kinase, and its activation enhances cell proliferation. Here, we report that ATF1 is a novel substrate of RSK2 and that RSK2-ATF1 signaling plays an important role in EGF-induced neoplastic cell transformation. RSK2 phosphorylated ATF1 at Ser-63 and enhanced ATF1 transcriptional activity. Docking experiments using the crystal structure of the RSK2 N-terminal kinase domain combined with in vitro pulldown assays demonstrated that eriodictyol, a flavanone found in fruits, bound with the N-terminal kinase domain of RSK2 to inhibit RSK2 N-terminal kinase activity. In cells, eriodictyol inhibited phosphorylation of ATF1 but had no effect on the phosphorylation of RSK, MEK1/2, ERK1/2, p38 or JNKs, indicating that eriodictyol specifically suppresses RSK2 signaling. Furthermore, eriodictyol inhibited RSK2-mediated ATF1 transactivation and tumor promoter-induced transformation of JB6 Cl41 cells. Eriodictyol or knockdown of RSK2 or ATF1 also suppressed Ras-mediated focus formation. Overall, these results indicate that RSK2-ATF1 signaling plays an important role in neoplastic cell transformation and that eriodictyol is a novel natural compound for suppressing RSK2 kinase activity. [ABSTRACT FROM AUTHOR]

• The chitosan with low molecular weight (1.20 × 104 Da) was prepared in this study. • TP5 showed strong antitumor effects and enhanced immunity on H22 tumor-bearing mice. • CS could suppress antitumor immune response of mice bearing tumor even under TP5 activation. • CS treatments could suppress the expressions of TNF-α, IFN-γ, IL-2 and IL-4 in sera of tumor-bearing mice. In the present study, the low molecular weight of chitosan (CS) was prepared and its activity on thymopentin-activated mice bearing H22 solid tumors was further researched. The purity and molecular weight of CS were determined by UV and HPGPC spectra, and its immunosuppressive effects on H22 tumor-bearing mice were evaluated through determination on immune organs, cells and cytokines. Results showed that CS contained little impurities with the average molecular weight of 1.20 × 104 Da. The in vivo antitumor experiments demonstrated that CS facilitated to destroy immune organs (thymuses and spleens), suppress immune cells (lymphocytes, macrophages and NK cells) activities and reduce immune-related cytokines (TNF-α, IFN-γ, IL-2 and IL-4) expressions of H22 tumor-bearing mice even with simultaneous TP5 stimulation. Our data suggested that CS could not be applied to improve immune response in cancer-bearing patients, but might be employed for treatments on autoimmune diseases or organ transplant patients. [ABSTRACT FROM AUTHOR]

Appropriate mechanical properties and highly interconnected porosity are important properties for tissue engineering scaffolds. However, most existing hydrogel scaffolds suffer from poor mechanical properties limiting their application. Furthermore, it is relatively infrequent that precision control is achieved over pore size and structure of the scaffold because there are relatively few current technologies that allow such control and there is not a general appreciation that such control is important. To address these shortcomings, by combining double network polymerization and sphere-templating fabrication techniques, we developed a tough, intelligent scaffold based on poly(acrylic acid) and poly(N-isopropyl acrylamide) with a controllable, uniform, and interconnected porous structure. A mechanical assessment showed the toughness of the hydrogel and scaffold to be up to ~1.4 × 107 Jm-3 and ~1.5 × 106 Jm-3 respectively, as compared with 104-105 Jm-3 for most synthetic hydrogels. The thermosensitivity and pH-sensitivity were explored in a swelling study. In vitro testing demonstrated the scaffold matrices supported NIH-3T3 cell adhesion, proliferation and infiltration. An in vivorabbit study showed the scaffolds promote strong cellular integration by allowing cells to migrate into the porous structure from the surrounding tissues. These data suggest that the poly(acrylic acid)/poly(N-isopropyl acrylamide)-based scaffold could be an attractive candidate for tissue engineering. [ABSTRACT FROM AUTHOR]

TRAF2 has an important function in mediating the TNF-R signaling pathway toward activation of NF-κB and JNKs. Here we reveal a novel function of TRAF2 in the epidermal growth factor (EGF) signaling pathway. Knockdown of TRAF2 blocked EGF-induced AP-1 activity and anchorage- independent cell transformation. Notably, we showed that EGF induces ribosomal S6 kinase 2 (RSK2) ubiquitination, and knocking down TRAF2 suppresses ubiquitination of RSK2 induced by EGF. We also found that TRAF2 affects RSK2 activity through RSK2 ubiquitination. RSK2 plays a critical role in AP-1 activity mediated through CREB and c-Fos, which regulates anchorage-independent cell transformation. In addition, TRAF2 is overexpressed in colon cancer and required for colon cancer development, suggesting that TRAF2 might be a potential molecular target for cancer prevention and treatment. [ABSTRACT FROM AUTHOR]

The ribosomal S6 kinase 2 (RSK2) is a member of the p90 ribosomal S6 kinase (p90RSK) family of proteins and plays a critical role in proliferation, cell cycle, and cell transformation. Here, we report that RSK2 phosphorylates caspase-8, and Thr-263 was identified as a novel caspase-8 phosphorylation site. In addition, we showed that EGF induces caspase-8 ubiquitination and degradation through the proteasome pathway, and phosphorylation of Thr-263 is associated with caspase-8 stability. Finally, RSK2 blocks Fas-induced apoptosis through its phosphorylation of caspase-8. These data provide a direct link between RSK2 and caspase-8 and identify a novel molecular mechanism for caspase-8 modulation by RSK2. [ABSTRACT FROM AUTHOR]

MST1 (mammalian STE20-like kinase 1) is a serine/threonine kinase that is cleaved and activated by caspases during apoptosis. Overexpression of MST1 induces apoptotic morphological changes such as chromatin condensation, but the mechanism is not clear. Here we show that MST1 induces apoptotic chromatin condensation through its phosphorylation of histone H2AX at Ser-139. During etoposide-induced apoptosis in Jurkat cells, the cleavage of MST1 directly corresponded with strong H2AX phosphorylation. In vitro kinase assay results showed that MST1 strongly phosphorylates histone H2AX. Western blot and kinase assay results with a mutant S139A H2AX confirmed that MST1 phosphorylates H2AX at Ser-139. Direct binding of MST1 and H2AX can be detected when co-expressed in HEK293 cells and was also confirmed by an endogenous immunoprecipitation study. When overexpressed in HeLa cells, both the MST1 full-length protein and the MST1 kinase domain (MST1-NT), but not the kinase-negative mutant (MST1- NT-KN), could induce obvious endogenous histone H2AX phosphorylation. The caspase-3 inhibitor benzyloxycarbonyl-DEVD-fluoromethyl ketone (Z-DEVD-fmk) attenuates phosphorylation of H2AX by MST1 but cannot inhibit MST1-NT-induced histone H2AX phosphorylation, indicating that cleaved MST1 is responsible for H2AX phosphorylation during apoptosis. Histone H2AX phosphorylation and DNA fragmentation were suppressed in MST1 knock-down Jurkat cells after etoposide treatment. Taken together, our data indicated that H2AX is a substrate of MST1, which functions to induce apoptotic chromatin condensation and DNA fragmentation. [ABSTRACT FROM AUTHOR]

Effective treatment of inflammatory pain is a major clinical concern for both patients and physicians. Traditional analgesics such as morphine and coxibs are not effective in all patients and have various unwanted side effects. Accumulating evidence has suggested that endomorphins (EMs), particularly EM-1, possess potent anti-inflammatory effects. However, poor bioavailability and low resistance to enzymatic degradation impede their direct application in the treatment of inflammation. A series of novel peptides based on the structure of EM-1, with lower undesired effects than their parent compounds, called MEL-EMs were discovered and synthetized in our preceding studies. Here, we selected two (MEL-0614 and MEL-N1606) to further investigate their anti-inflammatory effects. This work showed that MEL analogs exerted potent analgesic effects with the inhibition of activated glial cells and macrophages in a CFA-induced inflammatory pain model. Furthermore, multiple-dose administration of MEL analogs did not prolong CFA-induced chronic inflammatory pain, in contrast to morphine. Together, our findings revealed that MEL analogs may serve as effective candidates for chronic inflammation treatment. • MEL analogs relief the inflammatory pain induced by CFA. • MEL analogs inhibit the activation of non-neuronal cells, which is different from morphine. • Repetitive treatments of MEL analogs do not exaggerate inflammatory pain, but morphine has proinflammatory effects. • MEL analogs may not participate in spinal TLR4-NLRP3 pathway but morphine is involved. • MEL analogs reversed the prolongation and amplification induced by morphine. [ABSTRACT FROM AUTHOR]