Solution structure and p43 binding of the p38 leucine zipper motif: coiled-coil interactions mediate the association between p38 and p43

Abstractp38, which has been suggested to be a scaffold protein for the assembly of a macromolecular tRNA synthetase complex, contains a leucine zipper-like motif. To understand the importance of the leucine zipper-like motif of p38 (p38LZ) in macromolecular assembly, the p38LZ solution structure was investigated by circular dichroism and nuclear magnetic resonance spectroscopy. The solution structure of p38LZ showed an amphipathic α-helical structure and characteristics similar to a coiled-coil motif. The protein–protein interaction mediated by p38LZ was examined by an in vitro binding assay. The p43 protein, another non-synthetase component of the complex, could bind to p38LZ via its N-terminal domain, which is also predicted to have a potential coiled-coil motif. Thus, we propose that the p38–p43 complex would be formed by coiled-coil interactions, and the formation of the binary complex would facilitate the macromolecular assembly of aminoacyl-tRNA synthetases

Dietary isothiocyanate sulforaphene induces reactive oxygen species, caspase -9, -8, -3-dependent apoptosis and modulates PTEN/PI3Kinase in human cervical cancer cells

Purpose: To investigate the apoptotic activity, cell proliferation inhibition and different signaling protein expressions after treatment with a new isothiocyanate, sulforaphene, in human cervical cancer (HeLa) cells. Methods: Cytotoxicity was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay after sulforaphene treatment for 3, 6, 12 and 24 h. Apoptosis assay, cell cycle analysis, intracellular oxygen species (ROS) measurement, mitochondrial membrane depolarization and western blot analysis were performed in four time-intervals to explore sulforaphene activity. Results: HeLa cell viability was reduced by sulforaphene dose and time dependently. ROS plays a causative role in sulforaphene induced cytotoxicity and apoptosis during which stimulation of Bax and blocking of Bcl2 were involved. Mitochondrial membrane potential depletion and cytochrome C, AIF modulation suggest mitochondrial pathway for the apoptosis. Activation of caspase -9, -8 and -3 in treated HeLa cells demonstrated caspase-dependent apoptosis by sulforaphene. Again, sulforaphene induced HeLa cell proliferation inhibition was evidenced by cell cycle arrest and PTEN/PI3Kinase modulation. Conclusion: Dietary sulforaphene induces HeLa cell apoptosis by enhancing intracellular ROS levels, thereby activating multiple apoptotic signal cascades. Therefore, sulforaphene is a potential candidate for anticancer therapy. Keywords: Sulforaphene, HeLa cells, Apoptosis, ROS, Caspase activation, PTEN, PI3Kinas

Anti-oxidant and anti-inflammatory effects of rice bran and green tea fermentation mixture on lipopolysaccharideinduced RAW 264.7 macrophages

Purpose: To investigate the anti-inflammatory and anti-oxidant properties of an enzyme bath of Oryza sativa (rice bran) and Camellia sinensis O. Kuntz (green tea) fermented with Bacillus subtilis (OCB). Methods: The anti-oxidant effects of OCB were assessed by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assay and flow cytometry. The anti-inflammatory effects of OCB were assessed by a nitric oxide (NO) assay. Enzyme-linked immunosorbent assay and real-time polymerase chain reaction were used to quantify expression of pro-inflammatory cytokines in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The major compounds of OCB were identified using high performance liquid chromatography (HPLC) analysis. Results: OCB had no cytotoxic effect on LPS-stimulated macrophages or peripheral blood mononuclear cells up to 1 mg/mL. OCB displayed anti-oxidant effects comparable to those of ascorbic acid and reduced reactive oxygen species (ROS) levels in target cells. OCB treatment of LPSstimulated mavrophages decreased nitric oxide (NO), NO synthase (iNOS), cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2) and key pro-inflammatory cytokine expressions, suggesting that OCB acts as an anti-oxidant and anti-inflammatory agent by reducing ROS levels and inhibiting pro-inflammatory mediators. The main effector compounds in OCB were epicatechin gallate, cathechin, synigrin acid, epicathechin, epigallocatechin gallate, rutin, and isoquercetin, which are known anti-oxidants. Conclusion: OCB fermentation product may be used as synergistic adjuvant therapy for inflammatory diseases. Keywords: Rice bran, Green tea, Bacillus subtilis, Enzyme bath, Anti-oxidant, Anti-inflammatio

Sericea lespedeza (Lespedeza cuneata) whole plant extract enhances rat muscle mass and sperm production by increasing the activity of NO-cGMP pathway and serum testosterone

Purpose: To analyze the effects of an aqueous extract of Sericea lespedeza (SL) on rat male menopause.Methods: Levels of nitric oxide (NO), endothelial nitric oxide synthase (NOS), cGMP, and prostaglandin E2 (PGE2) in the penile corpus cavernosum of the rats were evaluated using appropriate kits. Serum levels of dihydrotestosterone (DHT), testosterone, sex hormone-binding globulin (SHBG), and 17-beta hydroxysteroid dehydrogenases (17β-HSD) were measured with enzyme-linked immunosorbent assay kits. Total and motile sperms were counted on a hemocytometer. Histological changes in rat testis and epididymis were analyzed with hematoxylin and eosin staining.Results: The levels of NO, NOS, and cGMP (but not PGE2) increased in a dose-dependent manner (p< 0.05) upon administration of an aqueous extract of SL (AESL), while levels of DHT, 17β-HSD, and testosterone increased in the group administered with 300 mg/kg of AESL. Epididymal sperm count increased by 24 % in such rats compared to controls (p < 0.05).Conclusion: The aqueous extract of SL improves sperm count and muscle mass in rats by increasing the levels of NO, NOS, cGMP and testosterone. Thus, SL extract can potentially be developed as an alternative therapeutic agent for clinical management of TDS. Keywords: NO-cGMP, Testosterone, Hormones, Sperm count, Muscle mass, Sericea lespedeza, Lespedeza cuneat

Three-dimensional nano-foam of few-layer graphene grown by CVD for DSSC

We report a robust and direct route to fabricate a three-dimensional nano-foam of few-layer graphene (3D-NFG) with large area coverage via a chemical vapor deposition (CVD) technique. Pyrolysis of polymer/nickel precursor film under a hydrogen environment, simply prepared by spin-coating, leads to the creation of nano-foam in the film and the reduction process of nickel ions. Carbonized-C and the nickel nano-frame formed from the pyrolysis are used as a solid carbon source and as a catalyst for the growth of graphene under CVD conditions, respectively. We investigate the use of 3D-NFG, with the advantage of large surface area and high conductivity, as an alternative to the Pt counter electrode material in dye sensitized solar cells. The excellent properties of 3D-NFG, fabricated in this simple and direct manner, suggest a great potential for interconnected graphene networks in electronic devices and photocatalytic sensors as well as in energy-related materials.close221

Functional Benefit after Modification of Radial Forearm Free Flap for Soft Palate Reconstruction

ObjectivesTo compare the velopharyngeal function, swallowing and speech of the conventional and modified radial forearm free flap (RFFF) for soft palate reconstruction.MethodsRetrospective clinical study. Twenty-eight patients who underwent oropharyngeal reconstruction with RFFF were divided into two groups: 10 patients had conventional folded RFFF and 18 patients underwent modified method.ResultsThe average speech intelligibility score in modified RFFF group was 8.0±2.4, and 6.2±2.2 in conventional RFFF group (P<0.05). The nasalance was 27.4±7.8% in modified group and 38.6±2.7% in conventional group during no nasal passage reading and 43.6±7.3% in modified group, 55.2±7.6% in conventional group during high nasal passage reading (P<0.05). The subjective swallowing functional score was 2.8 in modified group and 2.1 in conventional group.ConclusionThe speech assessment and nasalance demonstrate a more favorable outcome in modified group than conventional group

Fast track fed-batch culture development for COVID-19 vaccine clinical study

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