Proteins at the cell surface and within the endocytic pathway are increasingly being recognized for their roles in a wide variety of intercellular interactions. Here we used the inherent hydrophobicity and N-glycosylation of membrane proteins to enrich these proteins from the surface and endosome of avian LMH epithelial cells for mass spectrometric analysis. The cycling of many different types of proteins from the cell surface into the endosome and sometimes back to the surface again makes it appropriate to analyze these two membranous cellular components together. Stringent searches of the International Protein Index (IPI) entries for Gallus gallus identified 318 unique integral membrane proteins (IMPs) (201 bearing N-glycosylation sites), 265 unique membrane-associated proteins (MAPs), and an additional group of 784 non-membrane proteins (NMPs) among TX-114 detergent and aqueous phase-enriched proteins. Capture of N-glycosylated tryptic peptides revealed 36 additional glycoproteins most of which were CD antigens, receptors, and molecules for cell adhesion and immune response. IMPs and MAPs present at the surface and within the endosome included proteins involved in transport (255), metabolism (285), communication (108), adhesion (47), and immune responses (42). Among these were 355 putative uncharacterized and hypothetical IMPs, MAPs, and NMPs for which highly similar annotated sequences were found in standard protein-protein BLAST searches.Read more
To describe a modified suture technique for transperitoneal laparoscopic dismembered pyeloplasty (TPLDP) that can be consistently replicated.
Between June 2010 and April 2014, 21 men and 7 women with primary pelviureteric junction obstruction underwent our modified TPLDP suture technique performed by the same surgeon. In our method, the dismembering should be performed after performing half of anastomosis to achieve the maintenance of correct orientation and the prevention of torsion of anastomosis. We defined the success criteria as complete clinical resolution of flank pain for the patients with flank pain and complete radiologic resolution for the asymptomatic patients.
The mean overall operative time for our technique was 137.3 minutes. The mean operative time for procedures on the left side was longer than on the right side (P = .02). The mean suture time was 37.2 minutes. The mean estimated blood loss was 29.4 mL, and the crossing vessel was found in 7 of 28 patients (25.0%). No open conversion was required. The mean follow-up time was 21.0 months. Only 1 patient still had frequent and intolerable flank pain after surgery whose treatment was unsuccessful. The rest of the patients got complete clinical or radiologic resolution. The success rate was 27 of 28 (96.4%).
Our modified TPLDP suture technique is feasible and seems to be safe and to allow high success rate for the treatment of pelviureteric junction obstruction. Sizable sample and further analysis about technique will be completed in the future.Read more
Modification of the anoxic-oxic (AO) process by inserting a sludge holding tank (SHT) into the sludge return line forms an anoxic-oxic-settling-anaerobic (A+OSA) process that can achieve a 48.98% sludge reduction rate. The 454 pyrosequencing method was used to obtain the microbial communities of the AO and A+OSA processes. Results showed that the microbial community structures of the 2 processes were different as a result of the SHT insertion. Bacteria assigned to the phyla Proteobacteria and Bacteroidetes commonly existed and dominated the microbial populations of the 2 processes. However, the relative abundance of these populations shifted in the presence of SHT. The relative abundance of Proteobacteria decreased during the A+OSA process. A specific comparison at the class level showed that Sphingobacteria was enriched in the A+OSA process. The result suggested that the fermentative bacteria Sphingobacteria may have key functions in reducing the sludge from the A+OSA process. Uncultured Nitrosomonadaceae gradually became the dominant ammonia-oxidizing bacteria, and the nitrite-oxidizing bacterium Nitrospira was enriched in the A+OSA process. Both occurrences were favorable for stabilized nitrogen removal. The known denitrifying species in the A+OSA process were similar to those in the AO process; however, their relative abundance also decreased.Read more
With the rapid development of molecular diagnostic techniques, there is a growing need for quality controls and standards with favorable properties to monitor the entire detection process. In this study, we describe a novel method to produce armored hepatitis B virus (HBV) and human papillomavirus (HPV) DNA for use in nucleic acid tests, which was confirmed to be stable, homogeneous, noninfectious, nuclease resistant, and safe for shipping. We demonstrated that MS2 bacteriophage could successfully package double-stranded DNA of 1.3-, 3-, 3.5-, and 6.5-kb length into viral capsids with high reassembly efficiency. This is the first application of RNA bacteriophage MS2 as a platform to encapsulate double-stranded DNA, forming virus-like particles (VLPs) which were indistinguishable from native MS2 capsids in size and morphology. Moreover, by analyzing the interaction mechanism of pac site and the MS2 coat protein (CP), we found that in addition to the recognized initiation signal TR-RNA, TR-DNA can also trigger spontaneous reassembly of CP dimers, providing a more convenient and feasible method of assembly. In conclusion, this straightforward and reliable manufacturing approach makes armored DNA an ideal control and standard for use in clinical laboratory tests and diagnostics, possessing prospects for broad application, especially providing a new platform for the production of quality controls for DNA viruses.Read more
Concentrations of 17 polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and 12 dioxin-like polychlorinated biphenyls (dl-PCBs) were measured in 96 food composite samples from eight varieties of food groups from the Chinese total diet study (TDS) in 2007. The concentrations of samples, expressed as WHO toxic equivalents (TEQ), ranged from 0.001 pg TEQ g(-1) to 0.85 pg TEQ g(-1) (fresh weight). Dietary intake of PCDD/Fs and dl-PCBs of 12 age/gender subgroups of the Chinese population subsequently estimated ranges from 15.4 pg TEQ kg(-1) bw month(-1) to 38.7 pg TEQ kg(-1) bw month(-1) for average population and from 68.5 pg TEQ kg(-1) bw month(-1) to 226.1 pg TEQ kg(-1) bw month(-1) for high consumers (the 97.5th percentile). Dietary exposure of children (mean: 32.5 pg TEQ kg(-1) bw month(-1)) is significantly higher than that of the adults (mean: 21.5 pg TEQ kg(-1) bw month(-1)) (p<0.01) presumably due to more food consumed by children relative to their body weight compared to adults. There is no difference of dietary exposure, expressed as pg TEQ kg(-1) bw, found between different genders. Across various regions in China, there are large differences of dietary exposure of adult population and pattern of contribution of food groups to total exposure due to different contamination level and food habits. Dietary exposures of average population of various subgroups were all below the PTMI recommended by JECFA, but those of higher consumers were found exceeding or comparable to the PTMI.Read more
Emamectin benzoate (EMB), an important macrocyclic lactone insecticide that belongs to the avermectin family and possesses excellent potency in controlling pests, is non-carcinogenic and non-mutagenic conducted in rats and mice, but EMB-induced cytotoxicity and genotoxicity in arthropod insect have been seldom reported yet. In the present paper, we quantified the cytotoxicity of EMB through the detections on cell viability, DNA damage, and cell apoptosis in Spodoptera frugiperda Sf-9 cells in vitro. The results showed that EMB caused a concentration- and time-dependent reduction on the viability of Sf-9 cells, and the median inhibitory concentrations (IC50) were 3.34μM at 72h of exposure. The dual acridine orange/ethidium bromide staining showed that exposure to EMB induced a significant time- and concentration-dependent increase on cell apoptosis. The alkaline comet assay revealed that EMB induced significant increases on single-strand DNA breaks, and the percentage of γH2AX-positive cells represented a time- and concentration-dependent formation of DNA double-strand breaks in Sf-9 cells. Interestingly, the similar cytotoxic actions of EMB also went for the human cancerous HeLa cells as a control cell group. Data demonstrated the potential cytotoxic effect of EMB on Sf-9 cells that was significantly greater than the effect of hydrogen peroxide at the same concentrations.Read more
Nonferrous metal smelting is an important atmospheric mercury (Hg) emission source that has significant local and global impacts. To quantify the impact of Hg emission from non-ferrous metal smelter on the surrounding soil, an integrated model parameterizing the processes of smelter emission, air dispersion, atmospheric deposition and Hg accumulation in soil was developed. The concentrations of gaseous elemental Hg (GEM) around the smelter and the spatial distribution of Hg in the surrounding soil were measured and compared with the model results. Atmospheric deposition of Hg emitted from the smelter was identified as the main source of Hg accumulation in the surrounding soil. From 1960 to 2011, the smelter emitted approximately 105 t of Hg into the atmosphere, of which 15 t deposited locally and resulted in an increase of Hg concentration in soil from 0.12 to 1.77 mg kg(-1). A detailed examination of wind rose and model data suggested that the area within 1.0-1.5 km northwest and southeast of the smelter was most severely impacted. It was estimated that the smelter operation from 1969 to 1990, when large scale emission controls were not implemented, resulted in 6450 μg m(-2)yr(-1) of Hg net deposition and a model simulated increase of 0.40 mg kg(-1) of Hg accumulation in the soil. During the period from 1991 to 2011, atmospheric Hg emission from the smelter alone increased the average concentration in soil from 0.41 mg kg(-1) to 0.45 mg kg(-1). In the past 50 years, over 86% of Hg emitted from this smelter went into the global pool, indicating the importance of controlling Hg emissions from non-ferrous metal smelters.Read more
A simple and cheap method for depositing solution-processed GeO2 (sGeO2) film is proposed utilizing the weak solubility of GeO2 in water. X-ray photoelectron spectroscopy analysis reveals that a pure GeO2 thin film can be formed by casting its aqueous solution. This method can avoid the difficulty of vacuum evaporation by its high melting point. The sGeO2 film has been used successfully as an anode interfacial layer in poly(3-hexylthiophene) (P3HT) and indene-C60 bisadduct (IC60BA)-based bulk heterojunction organic solar cells with improved power conversion efficiency and device stability compared with that using conventional poly (3,4-ethylenedioxythiophene):poly (styrenesulfonate) (PEDOT:PSS); the improvement of the power conversion efficiency and the device stability are estimated to be 9% and 50%, respectively. The calculations of optical intensity in a whole cell demonstrate that a thin layer of sGeO2 could function as an optical spacer in the based bulk heterojunction (BHJ) organic solar cells (OSCs) for enhancing the light harvesting in the active layer. Interfacial evaluation by impedance spectroscopy shows that the sGeO2-based cell exists less charge carrier recombination and lower contact resistance. More importantly, the sGeO2 film processing is very simple and environmentally friendly, which has potential applications in green and low-cost organic electronics in the future.Read more
The surfactant assistant syntheses of sulfonic acid functionalized periodic mesoporous organosilicas with large pores are reported. A one-step condensation of tetramethoxysilane (TMOS) with 1,2-bis(trimethoxysilyl)ethane (BTME) and 3-mercaptopropyltrimethoxysilane (MPTMS) in highly acidic medium was performed in the presence of triblock copolymer Pluronic P123 and inorganic salt as additive. During the condensation process, thiol (-SH) group was in situ oxidized to sulfonic acid (-SO(3)H) by hydrogen peroxide (30 wt % H(2)O(2)). X-ray diffraction studies along with nitrogen and water sorption analyses reveal the formation of stable, highly hydrophobic, and well-ordered hexagonal mesoscopic structures in a wide range of -CH(2)CH(2)- concentrations in the mesoporous framework. The resultant materials were also investigated by (29)Si MAS and (13)C CP MAS NMR, thermogravimetric analyses, UV-Raman spectroscopy, and FT-IR spectroscopy. The role of the bridged organic group on the hydrothermal stability of the mesoporous materials was established, which revealed an enhancement in hydrothermal stability of the materials with incorporation of the bridged organic groups in the network. The catalytic performance of -SO(3)H functionalized mesoporous materials was investigated in the esterification of ethanol with acetic acid, and the results demonstrate that the ethane groups incorporated in the mesoporous framework have a positive influence on the catalytic behavior of the materials.Read more
Excessive iodide and fluoride coexist in the groundwater in many regions, causing a potential risk to the human thyroid. To investigate the mechanism of iodide- and fluoride-induced thyroid cytotoxicity, human thyroid follicular epithelial cells (Nthy-ori 3-1) were treated with different concentrations of potassium iodide (KI), with or without sodium fluoride (NaF). Cell morphology, viability, lactate dehydrogenase (LDH) leakage, apoptosis, and expression of inositol-requiring enzyme 1 (IRE1) pathway-related molecules were assessed. Results showed 50 mM of KI, 1 mM of NaF, and 50 mM of KI +1 mM of NaF changed cellular morphology, decreased viability, and increased LDH leakage and apoptosis. Elevated expression of binding protein (BiP), IRE1, and C/EBP homologous protein (CHOP) mRNA and protein, as well as spliced X-box-binding protein-1 (sXBP-1) mRNA, were observed in the 1 mM NaF and 50 mM KI +1 mM NaF groups. Collectively, excessive iodide and/or fluoride is cytotoxic to the human thyroid. Although these data do not manifest iodide could induce the IRE1 pathway, the cytotoxicity followed by exposure to fluoride alone or in combination with iodide may be related to IRE1 pathway-induced apoptosis. Furthermore, exposure to the combination of excessive iodide and fluoride may cause interactive effects on thyroid cytotoxicity.Read more