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- Author or Editor: W. Liu x
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Abstract
The glass transition of lyophilized materials is normally measured by conventional or temperature modulated differential scanning calorimetry (TMDSC). However, because of the weakness of these transitions when protein concentrations are high, these techniques are often unable to detect the glass transition (T g). High ramp rate DSC, where heating rates of 100 K per min and higher are used, has been shown to be able to detect weak transitions in a wide range of materials and has been applied to these materials in previous work. Dynamic mechanical analysis (DMA) is also known to be much more sensitive to the presence of relaxations in materials than other commonly used thermal techniques. The development of a method to handle powders in the DMA makes it now possible to apply this technique to protein and protein-excipient mixtures. HRR DSC, TMA and DMA were used to characterize the glass transition of lyophilized materials and the results correlated. DMA is shown to be a viable alternative to HRR DSC and TMA for lyophilized materials.
Abstract
Occupational exposure to Cr(VI) causes various effects including deep skin ulcerations. Its action mechanisms are not fully understood. In the present study, the evaluation of human dermal fibroblasts heat production was monitored, using microcalorimetry. as part of Cr(VI) toxicity. In control cells, normal heat production was 15±5 pW/cell. Regardless of the Cr(VI) concentration tested (0 to 500 μM), heat production was inhibited over time periods ranging from 3 to 25 h. These results could be correlated with cell mortality and the IC50 for Cr(VI) was 29±4 μM. In the WST-1 bioassay, the IC50 was 35±5 μM (no statistical difference). Thus, Cr(VI) altered the metabolism of the fibroblasts, and led to cellular death. Microcalorimetry can be a useful tool for determining the toxic effect of suspect compounds implicated in the occurrence of pathologies.
We present a new method to asses the strength of indirect interactions and to indentify candidate keystone species in quantitative food webs. We apply this method to the structural analysis of a host-parasitoid community. The strength and symmetry of indirect interactions between 12 leaf-miner hosts and their 27 hymenopteran parasitoids are quantified. It is shown that (1) quantifying longer pathways helps in determining which species have more important direct or indirect effects on others, (2) a keystone pattern of relative species importance, based on positionality in the interaction network, seems to characterize this community, (3) considering longer pathways results in a characteristic “few strong - many weak” distribution of interaction strength, and (4) between the majority of species pairs the interaction is weakly asymmetrical. We emphasise that a very simple network algebra approach may offer important predictions on both species- and community-level patterns.
Stem rust caused by Puccinia graminis f. sp tritici of wheat (Triticum aestivum L.) is one of the most destructive cereal diseases globally. Concern about the disease has increased since 1999 with the discovery in Uganda of a new virulent race of Pgt, designated as race TTKSK (also known as Ug99). The objectives of this experiment were to characterize the resistance and to determine the chromosomal location of the stem rust resistance in the spring wheat line PI 410966. A mapping population was developed from a cross between PI 410966 and a susceptible wheat line OK3040. An inoculation test with isolate 04KEN156/04 of race TTKSK was conducted at the USDA-ARS Cereal Disease Laboratory in the F6:7 generation, and the F6:7 phenotypic data were used to genetically map the resistance gene to the centromeric region on chromosome 2BS. The single locus explained the observed F6:7 resistant and susceptible scores. The location of the gene and molecular marker banding profiles of the diagnostic markers suggest that the stem rust resistance gene in PI 410966 could be a new gene, an allele of Sr36, or Sr36.
Wheat glutenins containing high and low molecular weight glutenin subunits (HMW-GS and LMW-GS) are the major determinants of wheat gluten quality. In this study, the recently developed reversed-phase ultra-performance liquid chromatography (RP-UPLC) was used to study the synthesis and accumulation patterns of glutenins during grain development of four Chinese bread wheat cultivars with different gluten quality. Developing grains were collected based on thermal times from 150 °Cd to 750 °Cd at 100 °Cd intervals, and the content of glutenin subunits and their accumulation patterns were determined by RP-UPLC as well as sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that HMW-GS and LMW-GS synthesis were initiated currently at 250 °Cd and they displayed a gradually upregulated expression. All the HMW-GS can be detected at 250 °Cd, earlier than LMW-GS. Different glutenin subunits and genotypes showed clear accumulation diversity during grain development. Particularly, 1Dx5 + 1Dy10 in the cultivar Gaocheng 8901 and Zhongyou 9507 with superior dough properties were accumulated faster at early stages than 1Dx2 + 1Dy12 in Jingdong 8 and Zhengmai 9023 with poor dough quality, suggesting that faster accumulation rate of glutenin proteins at the early stages of grain development may contribute to the formation of superior gluten structure and dough quality.
Shenqi Fuzheng Injection (SFI) is a traditional Chinese medicine injection, widely used to enhance immune function of clinical cancer patients undergoing chemotherapy. In this study, a high-performance liquid chromatography-diode array detection-evaporative light scattering detection (HPLC-DAD-ELSD) method was established for quality control of SFI, which could simultaneously semiquantitatively reflect the constituents displayed in the chromatographic profile of SFI. The relative retention time and relative peak areas of the 21 common peaks related to the reference peak were calculated. The validity and advantage of this method were validated by systematically comparing chromatograms of 10 batches of SFI samples with the analytical methods of principal component analysis and angle cosine method recommended by the State Food and Drug Administration of China. Moreover, a total of 21 constituents of SFI were identified or tentatively characterized in the fingerprint via ultrafast liquid chromatography-diode array detection-quadrupole time-of-flight (UFLC-DAD-Q-TOF) tandem mass spectrometry technique on the basis of the retention time, ultraviolet spectra, fragmentation patterns, and reported literatures. All the results proved that the technique was useful in comprehensive quality evaluation of SFI and further study.
Abstract
The extraction of cobalt by Winsor II microemulsion system was studied. In the bis (2-ethylhexyl) sulfosuccinate sodium salt (AOT)/n-pentanol/n-heptane/NaCl system, AOT was used as a anionic surfactant to form microemulsion in n-heptane, n-pentanol was injected in the microemulsion as a cosurfactant. Co(II) was found to be extracted into the microemulsion phase due to ion pair formation such as Co2+(R–SO3 −)Cl. The influence of different parameters such as the volume ratio of aqueous phase to microemulsion, surfactant concentration, pH of the feed solutions, cosurfactant concentration as well as temperature on the extraction yield (E%) were investigated. The results showed that it was possible to extract 95% of cobalt by the AOT Winsor II microemulsion.
In this study, the cDNA of homocysteine S-methyltransferase was isolated from Aegilops tauschii Coss., with the gene accordingly designated as AetHMT1. Similar to other methyltransferases, AetHMT1 contains a GGCCR consensus sequence for a possible zinc-binding motif near the C-terminal and a conserved cysteine residue upstream of the zinc-binding motif. Analysis of AetHMT1 uncovered no obvious chloroplast or mitochondrial targeting sequences. We functionally expressed AetHMT1 in Escherichia coli and confirmed its biological activity, as evidenced by a positive HMT enzyme activity of 164.516 ± 17.378 nmol min−1 mg−1 protein when catalyzing the transformation of L-homocysteine. Compared with the bacterium containing the empty vector, E. coli harboring the recombinant AetHMT1 plasmid showed much higher tolerance to selenate and selenite. AetHMT1 transcript amounts in different organs were increased by Na2SeO4 treatment, with roots accumulating higher amounts than stems, old leaves and new leaves. We have therefore successfully isolated HMT1 from Ae. tauschii and characterized the biochemical and physiological functions of the corresponding protein.
Abstract
Although the use of aspirin has substantially reduced the risks of cardiovascular events and death, its potential mechanisms have not been fully elucidated. In a previous study, we found that aspirin triggers cellular autophagy. In the present study, we aimed to determine the protective effects of aspirin on human coronary artery endothelial cells (HCAECs) and explore its underlying mechanisms. HCAECs were treated with oxidized low-density lipoprotein (ox-LDL), angiotensin II (Ang-II), or high glucose (HG) with or without aspirin stimulation. The expression levels of endothelial nitric oxide (NO) synthase (eNOS), p-eNOS, LC3, p62, phosphor-nuclear factor kappa B (p-NF-κB), p-p38 mitogen-activated protein kinase (p-p38 MAPK), and Beclin-1 were detected via immunoblotting analysis. Concentrations of soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1) were measured via ELISA. NO levels were determined using the Griess reagent. Autophagic flux was tracked by tandem mRFP-GFP-tagged LC3. Results showed that aspirin increased eNOS level and reduced injury to the endothelial cells (ECs) caused by ox-LDL, Ang-II, and HG treatment in a dose-dependent manner. Aspirin also increased the LC3II/LC3I ratio, decreased p62 expression, and enhanced autophagic flux (autophagosome and autolysosome puncta) in the HCAECs. p-NF-κB and p-p38 mitogen-activated protein kinase inhibition, sVCAM-1 and sICAM-1 secretion, and eNOS activity promotion by aspirin treatment were found to be dependent on Beclin-1. These results suggested that aspirin can protect ECs from ox-LDL-, Ang-II-, and HG-induced injury by activating autophagy in a Beclin-1-dependent manner.
Abstract
Dodecylamine hydrochloride C12H25NH3·Cl(s) and bis-dodecylammonium tetrachlorozincate (C12H25NH3)2ZnCl4(s) were synthesized by the method of liquid phase reaction. The constant-volume energy of combustion of dodecylamine hydrochloride was measured by means of a RBC-II precision rotating-bomb combustion calorimeter at T = (298.15 ± 0.001) K. The standard molar enthalpy of formation of C12H25NH3·Cl(s) was calculated to be 
(C12H25NH3·Cl, s) = −(706.79 ± 3.97) kJ mol−1 from the constant-volume energy of combustion. In accordance with Hess’ law, a reasonable thermochemical cycle was designed and the enthalpy change of the synthesis reaction of the complex (C12H25NH3)2ZnCl4(s) was determined by use of an isoperibol solution-reaction calorimeter. The standard molar enthalpy of formation of (C12H25NH3)2ZnCl4(s) was calculated as 
[(C12H25NH3)2ZnCl4, s] = −(1862.14 ± 7.95) kJ mol−1 from the standard molar enthalpy of formation of C12H25NH3·Cl(s) and other auxiliary thermodynamic data.
