Danielsenfagan1885

Proper glutamatergic neurotransmission requires a balance between glutamate release and removal. The removal is mainly catalyzed by the glutamate transporters EAAT1-3, while the glutamate-cystine exchanger (system xc- with specific subunit xCT) represents one of the release mechanisms. Previous studies of the spinal cord have focused on the cellular distribution of EAAT1-3 with special reference to the dorsal horn, but have not provided quantitative data and have not systematically compared multiple segments. Here we have studied the distribution of EAAT1-3 and xCT in sections of multiple spinal cord segments using knockout tissue as negative controls. EAAT2 and EAAT3 were evenly expressed in all gray matter areas at all segmental levels, albeit with slightly higher levels in laminae 1-4 (dorsal horn). Somewhat higher levels of EAAT2 were also seen in lamina 9 (ventral horn), while EAAT3 was also detected in the lateral spinal nucleus. EAAT1 was concentrated in laminae 1-3, lamina 10, the intermediolateral nucleus and the sacral parasympathetic nucleus, while xCT was concentrated in laminae 1-3, lamina 10 and the leptomeninges. The levels of these four transporters were low in white matter, which represents 42% of the spinal cord volume. Quantitative immunoblotting revealed that the average level of EAAT1 in the whole spinal cord was 0.6 ± 0.1% of that in the cerebellum, while the levels of EAAT2, EAAT3 and xCT were, respectively, 41.6 ± 12%, 39.8 ± 7.6%, and 30.8 ± 4.3% of the levels in the hippocampus (mean values ± SEM). Conclusions Because the hippocampal tissue content of EAAT2 protein is two orders of magnitude higher than the content of the EAAT3, it follows that most of the gray matter in the spinal cord depends almost exclusively on EAAT2 for glutamate removal, while the lamina involved in the processing of autonomic and nociceptive information rely on a complex system of transporters.Pullulan, a naturally occurring polysaccharide was oxidized using sodium periodate and converted into its aldehyde derivative and used as a bio crosslinker for stabilizing collagen. The crosslinking mainly occurred due to Schiff's base reaction between amino group of lysine or hydroxylysine present in collagen with the aldehyde groups of oxidized pullulan. A known concentration of collagen was crosslinked with different concentrations of oxidized pullulan and scaffolds were prepared by freeze drying method. The crosslinking efficiency and biodegradation of the crosslinked scaffolds showed higher degree of crosslinking and slower degradation rate with increase in cross linker concentration. The surface morphology of scaffolds using SEM showed that the scaffolds with 3% concentration of crosslinker (Col-OxP-3) exhibited highly porous nature with interconnecting pores of 66.35 μm size compared to other groups, which was also supported by porosity and tensile strength measurement. The tensile strength was 10 times higher in Col-OxP-3 scaffold compared to native collagen. The swelling capacity of the scaffolds increased with increase in crosslinker concentration and the equilibrium was retained up to 60 min. The crosslinked scaffolds were non-toxic to 3T3 fibroblast cell line proving their biocompatible nature. Thus, this new class of bio-crosslinker can be used in replacement of synthetic crosslinkers for biomaterial synthesis.The search for new approaches for developing antimicrobial surfaces is a challenge of great urgency to prevent and control microbial growth on surfaces. The strategy herein proposed relies on the design of a new, simple and general tool for creating antimicrobial surfaces, based on a hydrophobin chimeric protein which fuses the adhesive self-assembling class I hydrophobin Vmh2 from Pleurotus ostreatus to the human antimicrobial peptide LL-37. The recombinant LL37-Vmh2 protein displayed both the adhesive and the antimicrobic properties of its members, and when deposited on polystyrene surface, a positive effect due to the fusion was observed in terms of both efficacy and versatility of the coating. Indeed, the chimeric protein significantly enlarges the range of pathogens affected by Vmh2 layer rendering it able to inhibit three Gram-positive and two Gram-negative pathogens, selected among the renowned biofilm producer bacteria. Confocal Laser Scanning Microscopy analysis performed on Staphylococcus epidermidis biofilms formed on coated surfaces proved that, besides inhibiting biofilm formation, the LL37-Vmh2 coating also displayed biocidal activity, since dead cells were present in the biofilm layer. The reported results open new perspectives in various fields of application of LL37, and of antimicrobial peptides in general. LL37-Vmh2 increases the inventory of chimeric hydrophobins, further proving their effectiveness and versatility in surface functionalization.Trop2 is an intracellular calcium signal transducer and a prognostic biomarker in many cancers. P16 is a cell cycle gene that negatively regulates cell proliferation and division in most human cancers. Oral squamous cell carcinoma (OSCC) is a common malignant tumor subgroup of head and neck squamous cell carcinoma worldwide. Both Ca2+-dependent and cell cycle signaling pathways play vital roles in OSCC, although the molecular mechanisms remain to be elucidated. We aimed to examine the function of Trop2 and P16 in regulating intracellular calcium ions and the cell cycle in OSCC cell lines. Furtherly, the mRNA and protein expression levels of Trop2 and P16 in OSCC tissue samples were assessed, and their function was evaluated as potential clinical prognostic biomarkers. Trop2 promoted intracellular calcium ion release in OSCC and induced S phase of the cell cycle. Moreover, Trop2-mediated Ca2+ inhibited P16 expression through the AMP-activated protein kinase pathway in OSCC. Interestingly, P16 overexpression could not reverse these phenomena in vitro. selleck chemical We also demonstrated that human OSCC tissues showed high Trop2 mRNA and protein expression, and Trop2+/P16- expression is an independent prognostic marker for OSCC patients. Our data suggest that Trop2+/P16- may be a valuable prognostic marker for OSCC and that Trop2 inhibits P16 expression and induces S phase by promoting intracellular calcium release in OSCC.Sulfated polysaccharides were shown to benefit metabolic syndrome (MS) and gut microbiota, but the contribution of sulfate group remains unclear. In this study, sulfated polysaccharides from pacific abalone (AGSP) and its desulfated product (D-AGSP) were prepared, and the contribution of sulfate group was analyzed via in vitro and in vivo models. The result showed that sulfate group had no obvious effect on the reaction of AGSP with RAW 264.7 cells, but it affected the growth properties of gut microbes that able to utilize AGSP. The mice experiment showed that D-AGSP reduced weight gain, fat accumulation and lipid metabolism disorder in HFD-fed mice as well as AGSP, and no differences between them were found. Sequencing analysis showed that sulfate group influenced AGSP-induced alterations of the gut microbiota at higher taxonomic levels, some of which had close correlation with the improvement of physiological index. These results implied that sulfate group may partially determine the activities of polysaccharides via gut microbiota-mediated pathway, but the exact mechanisms need further research.This study undertakes the development of colloidal carriers for the purpose of oral delivery of bosentan and subsequent management of systemic hypertension. Karaya gum, a natural polymer was carboxymethylated to improve its hydrophilic character and then the carboxymethyl gum was hydrophobically modified by forming propyl ethers. The modified polymer acquired amphiphilic property and self-aggregated in water to form amphiphilic colloidal particles (ACPs) at critical concentration of 3.35 mg/L with spherical shape (90% drug in the lipophilic domain. The ionic crosslinking of the hydrophilic shell of ACPs imparted greater stability to the colloidal system. The crosslinking extended the duration of drug release under simulated gastrointestinal fluids. The crystalline drug physically turned into amorphous state after hosting into the lipophilic cores of ACPs. The entrapment resulted in significant improvement of drug dissolution rate. The polymer relaxation contributed to the diffusion process of drug from ACPs. Pre-clinical testing via oral route demonstrated that the crosslinked colloidal particles could effectively control the systemic hypertension over a period of 12 h. Hence, bosentan-loaded self-assembled colloidal particles may advance the management of systemic hypertension.Hydrogel wound dressing is a type of hydrophilic polymer, which has been widely studied and applied in biomedical field. In this study, a simple and non-toxic method was developed to prepare a new type of composite hydrogel, which was formed through the Schiff-base reaction between the aldehyde of Oxidized Hydroxyethyl Cellulose (OHEC) and the amino of Carboxymethyl Chitosan (CMCS). Hence, a series of tests toward this new composite hydrogel which contained its structure and performance was applied. Statistics achieved from those tests showed that this composite hydrogel comprised of some high-quality properties such as suitable gelation time, good swelling ability, suitable water evaporation rate, good blood compatibility and biocompatibility. Considering these properties, this hydrogel has a potential to be explored as wound dressing.The acyl-CoA dehydrogenase (FadE) and (R)-specific enoyl-CoA hydratase (PhaJ) are functionally related to the degradation of fatty acids and the synthesis of polyhydroxyalkanoates (PHAs). To verify this, a recombinant Cupriavidus necator H16 harboring the plasmid -pMPJAS03- with fadE from Escherichia coli strain K12 and phaJ1 from Pseudomonas putida strain KT2440 under the arabinose promoter (araC-PBAD) was constructed. The impact of co-expressing fadE and phaJ genes on C. necator H16/pMPJAS03 maintaining the wild-type synthase on short-chain-length/medium-chain-length PHA formation from canola or avocado oil at different arabinose concentrations was investigated. The functional activity of fadEE.c led to obtaining higher biomass and PHA concentrations compared to the cultures without expressing the gene. While high transcriptional levels of phaJ1P.p, at 0.1% of arabinose, aid the wild-type synthase to polymerize larger-side chain monomers, such as 3-Hydroxyoctanoate (3HO) and 3-Hydroxydecanoate (3HD). The presence of even small amounts of 3HO and 3HD in the co-polymers significantly depresses the melting temperature of the polymers, compared to those composed of pure 3-hydroxybutyrate (3HB). Our data presents supporting evidence that the synthesis of larger-side chain monomers by the recombinant strain relies not only upon the affinity of the wild-type synthase but also on the functionality of the intermediate supplying enzymes.Zeolite-Mg/Fe chloride dual enhanced coagulation is a cost-effective method for advanced treatment of swine wastewater, but the sludge generated after the enhanced coagulation remains to be a problem. In this study, the precipitate from a swine wastewater coagulation unit was regenerated by pyrolysis treatment in an O2-limited environment to develop a high efficient adsorbent (biochar-mineral composite, BMC) for the removal of Pb(II) from wastewater. SEM images indicate that complex Mg/Fe oxides and sludge biochar gathered around zeolite particles. Effects of different influencing factors such as Pb(II) initial concentration, pH, adsorption time and ion concentration on the adsorption performance were investigated. The results show that the Langmuir isotherm model can better express the adsorption of Pb(II) on BMC than Freundlich model and Temkin model. BMC pyrolyzed at 500 °C showed the maximum adsorption capacity of 450.58 mg/g under experimental condition of 25 °C, 100 mg/L Pb(II) initial concentration and the initial pH of 5.