Human serum albumin (HSA) has been widely used as a pharmaceutical excipient in Botulinum toxin serotype A (BoNT/A) products that are indicated for use in therapeutics and cosmetics. However, HSA as ...a human-derived material has some concerns, such as the potential risk of transmission of infectious agents, an insufficient supply, and difficulty in maintaining a certain quality. For those reasons, newly developed BoNT/A products (CORETOX®, Medytox, Inc., Republic of Korea) contained polysorbate 20, a non-human-derived excipient, to replace the HSA. However, most safety studies of polysorbate 20 have been conducted with non-invasive routes of administration, and thus there are a few studies on the safety of polysorbate 20 when administered intramuscularly. To secure the in vivo safety profile of polysorbate 20, a four-week repeated intramuscular dose toxicity study (0.02, 0.1, and 0.4 mg/kg, one injection every two weeks for a total of three injections) was conducted in 66 Sprague-Dawley (SD) rats. An intradermal irritation study was further conducted with 18 New Zealand White (NZW) rabbits. The toxicological evaluation of HSA (0.06 and 0.12 mg/kg) was also carried out as a comparative substance. Systemic and local toxicities were not observed in any of the SD rats or NZW rabbits based on clinical signs, body weight, hematology, clinical biochemistry, macroscopic findings on necropsy, histopathology of the injection site, and allergic reactions. The current study suggested that intramuscular administration of polysorbate 20 was considered to be safe at a level similar to that of HSA, which has an in vivo safety profile accumulated over the years. This provided the basis for the in vivo safety profile of polysorbate 20 administered intramuscularly and the scientific reliability of the use of polysorbate 20 as an alternative to HSA, which is used as an excipient for various pharmaceuticals in terms of its safety.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Sensory axons degenerate following separation from their cell body, but partial injury to peripheral nerves may leave the integrity of damaged axons preserved. We show that an endogenous ligand for ...the natural killer (NK) cell receptor NKG2D, Retinoic Acid Early 1 (RAE1), is re-expressed in adult dorsal root ganglion neurons following peripheral nerve injury, triggering selective degeneration of injured axons. Infiltration of cytotoxic NK cells into the sciatic nerve by extravasation occurs within 3 days following crush injury. Using a combination of genetic cell ablation and cytokine-antibody complex stimulation, we show that NK cell function correlates with loss of sensation due to degeneration of injured afferents and reduced incidence of post-injury hypersensitivity. This neuro-immune mechanism of selective NK cell-mediated degeneration of damaged but intact sensory axons complements Wallerian degeneration and suggests the therapeutic potential of modulating NK cell function to resolve painful neuropathy through the clearance of partially damaged nerves.
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•Cytotoxic NK cells infiltrate the damaged peripheral nerve within days of injury•Injured sensory axons express NKG2D ligand RAE1 to signal degeneration by NK cells•Clearance of damaged axons reduces development of chronic pain after nerve injury•NK cells complement Wallerian degeneration to aid functional regeneration of PNS
Pain following nerve damage can be mitigated by cytotoxic natural killer cells that selectively clear out partially damaged nerves.
Hot electrons and surface-plasmon-driven chemistry are amongst the most actively studied research subjects because they are deeply associated with energy dissipation and the conversion processes at ...the surface and interfaces, which are still open questions and key issues in the surface science community. In this topical review, we give an overview of the concept of hot electrons or surface-plasmon-mediated hot electrons generated under various structural schemes (i.e. metals, metal-semiconductor, and metal-insulator-metal) and their role affecting catalytic activity in chemical reactions. We highlight recent studies on the relation between hot electrons and catalytic activity on metallic surfaces. We discuss possible mechanisms for how hot electrons participate in chemical reactions. We also introduce controlled chemistry to describe specific pathways for selectivity control in catalysis on metal nanoparticles.
Biliary atresia (BA) results in severe bile blockage and is caused by the absence of extrahepatic ducts. Even after successful hepatic portoenterostomy, a considerable number of patients are likely ...to show progressive deterioration in liver function. Recent studies show that mutations in protein-coding mitochondrial DNA (mtDNA) genes and/or mitochondrial genes in nuclear DNA (nDNA) are associated with hepatocellular dysfunction. This observation led us to investigate whether hepatic dysfunctions in BA is genetically associated with mtDNA mutations. We sequenced the mtDNA protein-coding genes in 14 liver specimens from 14 patients with BA and 5 liver specimens from 5 patients with choledochal cyst using next-generation sequencing. We found 34 common non-synonymous variations in mtDNA protein-coding genes in all patients examined. A systematic 3D structural analysis revealed the presence of several single nucleotide polymorphism-like mutations in critical regions of complexes I to V, that are involved in subunit assembly, proton-pumping activity, and/or supercomplex formation. The parameters of chronic hepatic injury and liver dysfunction in BA patients were also significantly correlated with the extent of hepatic failure, suggesting that the mtDNA mutations may aggravate hepatopathy. Therefore, mitochondrial mutations may underlie the pathological mechanisms associated with BA.
To evaluate the retinal microvasculature of peripapillary and macular regions in highly myopic eyes and investigate the association between the vascular parameters and optic nerve head (ONH) ...deformation.
Cross-sectional study.
Seventy-one subjects with highly myopic eyes and 26 subjects with emmetropic eyes were included. Horizontal B-scan images of the ONH were obtained using optical coherence tomography (OCT) and horizontal tilt angles were measured. Integrated automated algorithms in the Avanti OCT angiography were used to quantify the peripapillary vessel density and area of the foveal avascular zone (FAZ) at the level of superficial and deep vascular networks. Association between horizontal disc tilt and the vascular parameters was evaluated.
The mean axial length and horizontal tilt angle were 26.73 ± 0.63 mm and 9.77 ± 3.00 degrees in the highly myopic group and 23.46 ± 0.55 mm and 5.95 ± 3.48 degrees in the emmetropic group, respectively. Highly myopic eyes exhibited significantly lower average peripapillary vessel density (P = .010) and larger superficial and deep FAZs (P = .001 and P < .001, respectively) compared with emmetropic eyes. Linear regression analyses showed that horizontal tilt angle significantly correlated the average peripapillary vessel density (P = .037) and the areas of superficial (P < .001) and deep (P < .001) FAZs.
The retinal microvasculature was significantly different in highly myopic eyes according to the degree of horizontal optic disc tilt. In addition to peripapillary vessel density, FAZ can be affected by the degree of optic disc tilt.
Biodiesel production using oleaginous yeasts, such as Yarrowia lipolytica, has recently attracted strong attention in terms of lipid yield and productivity. However, efficient methods of lipid ...extraction from oleaginous yeasts are required to further improve the economic feasibility of yeast-based biodiesel production. We investigated the effectiveness of selected switchable hydrophilicity solvents (SHSs), namely, N-ethylbutylamine (EB), N-dipropylamine (DP), and N,N-dimethylcyclohexyl-amine (DMCHA), for lipid extraction from the oleaginous yeast Y. lipolytica. Regardless of using harvested cells or culture broth, DMCHA and EB effectively extracted lipids from Y. lipolytica, resulting in up to 13% higher lipid yields than that obtained by conventional organic solvent extraction using a chloroform/methanol mixture. The lipid extracts obtained by the SHSs showed fatty acid compositions comparable to that obtained with the conventional solvent, thus assuring the quality of biodiesel produced through this method. After switching processes by purging with CO2 and N2, the lipids and solvents were successfully recovered, demonstrating the practicability of lipid extraction using SHSs. Consequently, this study provides an effective lipid extraction method for Y. lipolytica that could significantly improve energy efficiency and economic feasibility of an oleaginous-yeast-based biodiesel production process.
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•Lipids were extracted from Yarrowia lipolytica by SHSs (DMCHA, EB, and DP).•DMCHA and EB showed up to 13% higher yields relative to the conventional method.•Lipids were effectively extracted from both harvested wet cells and culture broth.•This extraction method would improve energy efficiency and economic feasibility.
The construction of microbial cell factories requires cost-effective and rapid strain development through metabolic engineering. Recently, RNA-guided CRISPR technologies have been developed for ...metabolic engineering of industrially-relevant host.
To demonstrate the application of the CRISPR interference (CRISPRi), we developed two-plasmid CRISPRi vectors and applied the CRISPRi in Corynebacterium glutamicum to repress single target genes and double target genes simultaneously. Four-different single genes (the pyc, gltA, idsA, and glgC genes) repressions were successfully performed using the CRISPRi vectors, resulting significant mRNA reductions of the targets compared to a control. Subsequently, the phenotypes for the target gene-repressed strains were analyzed, showing the expected cell growth behaviors with different carbon sources. In addition, double gene repression (the idsA and glgC genes in a different order) by the CRISPRi resulted in an independent gene repression to each target gene simultaneously. To demonstrate an industrial application of the CRISPRi, citrate synthase (CS)-targeting DM1919 (L-lysine producer) strains with a sgRNA-gltA-r showed reduced CS activity, resulting in the improvement of L-lysine yield by 1.39-fold than the parental DM1919 (a lysine producer).
Single or double gene repression were successfully performed using the CRISPRi vectors and sequence specific sgRNAs. The CRISPRi can be applied for multiplex metabolic engineering to enhanced lysine production and it will promote the further rapid development of microbial cell factories of C. glutamicum.
Abstract
We investigated the effects of using a virtual reality smartphone-based head-mounted display (VR SHMD) device for 2 h on visual parameters. Fifty-eight healthy volunteers were recruited. The ...participants played games using VR SHMD or smartphones for 2 h on different days. Visual parameters including refraction, accommodation, convergence, stereopsis, and ocular alignment and measured choroidal thickness before and after the use of VR SHMD or smartphones were investigated. Subjective symptoms were assessed using questionnaires. We analyzed the differences in visual parameters before and after the use of VR SHMD or smartphones and correlations between baseline visual parameters and those after the use of the devices. Significant changes were observed in near-point convergence and accommodation, exophoric deviation, stereopsis, and accommodative lag after the use of VR SHMD but not after that of smartphones. The subjective discomfort associated with dry eye and neurologic symptoms were more severe in the VR group than in the smartphone group. There were no significant changes in refraction and choroidal thickness after the use of either of the two devices. The poorer the participants’ accommodation and convergence ability the greater the resistance to changes in these visual parameters, and participants with a large exophoria were more prone to worsening of exophoria than those with a small exophoria.
Kidney disability due to kidney failure could be considered to be the most severe of all the internal-organ disabilities. The purpose of this study was to identify the disease burden between the ...kidney and non-kidney disabled among the internal-organ disabled, based on the number of chronic diseases, annual out-of-pocket expenditure, and quality of life. From 2009 to 2013, 308 people (6.5%) with internal-organ disabilities were extracted out of 4732 people with disabilities in the Korea Health Panel. We compared the disease burden of 136 people with kidney disability (44.2%) and 172 people with non-kidney disability (55.8%), and confirmed the trend of disease burden over five years through panel analysis. The disease burden gap between kidney and non-kidney disabilities was, respectively, the number of chronic diseases (4.7 vs. 3.3,
< 0.0001), annual out-of-pocket expenditure ($1292 vs. $847,
< 0.004), and quality of life score out of 100 (49.2 vs. 60.2,
< 0.0001). In addition, when looking at the five-year trend of the three disease burden indexes, the kidney disabled were consistently worse than the non-kidney disabled (
< 0.01). In conclusion, health policy planners aiming for health equity need to seek practical strategies to reduce the gap in the disease burden among people with disabilities.
Transforming growth factor-β (TGF-β) promotes tumor invasion and metastasis by inducing epithelial-mesenchymal transition (EMT). EMT is often related with acquisition of stemness characteristics. The ...objective of this study was to determine whether EMT and stemness characteristics induced by TGF-β might be associated with epigenetic regulation in lung cancer. A human normal lung epithelial cell line and four lung cancer cell lines were treated with TGF-β. Transcriptome analysis of BEAS-2B and A549 cells incubated with TGF-β were analyzed through next-generation sequencing (NGS). Western blotting was carried out to investigate expression levels of epithelial and mesenchymal markers. Wound healing and Matrigel invasion assay, sphere formation assay, and in vivo mice tumor model were performed to evaluate functional characteristics of EMT and stemness acquisition. To investigate whether activation of EMT and stem cell markers might be involved in epigenetic regulation of lung cancer, experiment using a DNA methyltransferase inhibitor (5-azacytidine, AZA), methylation-specific PCR (MSP) and bisulfite sequencing were performed. NGS revealed changes in expression levels of EMT markers (E-cadherin, N-cadherin, fibronectin, vimentin, slug and snail) and stem cell markers (CD44 and CD87) in both BEAS-2B and A549 cells. Functional analysis revealed increased migration, invasion, sphere formation, and tumor development in mice after TGF-β treatment. Expression of slug and CD87 genes was activated following treatment with AZA and TGF-β. MSP and bisulfite sequencing indicated DNA demethylation of slug and CD87 genes. These results suggest that TGF-β induced EMT and cancer stemness acquisition could be associated with activation of slug and CD87 gene by their promoter demethylation.
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