Study of the Sensing Kinetics of G Protein-Coupled Estrogen Receptor Sensors for Common Estrogens and Estrogen Analogs.

Endogenous and exogenous estrogens are widely present in food and food packaging, and high levels of natural estrogens and the misuse or illegal use of synthetic estrogens can lead to endocrine disorders and even cancer in humans. Therefore, it is consequently important to accurately evaluate the presence of food-functional ingredients or toxins with estrogen-like effects. In this study, an electrochemical sensor based on G protein-coupled estrogen receptors (GPERs) was fabricated by self-assembly, modified by double-layered gold nanoparticles, and used to measure the sensing kinetics for five GPER ligands. The interconnected allosteric constants (Ka) of the sensor for 17ß-estradiol, resveratrol, G-1, G-15, and bisphenol A were 8.90 × 10-17, 8.35 × 10-16, 8.00 × 10-15, 5.01 × 10-15, and 6.65 × 10-16 mol/L, respectively. The sensitivity of the sensor for the five ligands followed the order of 17ß-estradiol > bisphenol A > resveratrol > G-15 > G-1. The receptor sensor also demonstrated higher sensor sensitivity for natural estrogens than exogenous estrogens. The results of molecular simulation docking showed that the residues Arg, Glu, His, and Asn of GPER mainly formed hydrogen bonds with -OH, C-O-C, or -NH-. In this study, simulating the intracellular receptor signaling cascade with an electrochemical signal amplification system enabled us to directly measure GPER-ligand interactions and explore the kinetics after the self-assembly of GPERs on a biosensor. This study also provides a novel platform for the accurate functional evaluation of food-functional components and toxins.

Critical roles of the E3 ubiquitin ligase FBW7 in B-cell response and the pathogenesis of experimental autoimmune arthritis.

Proper regulation of B-cell function is essential for effective humoral immunity and maintenance of immune tolerance. Here, we found that FBW7 (F-box/WD40 repeat-containing protein 7) is highly expressed in germinal centre B and B1 cells, and confirmed that it has an intrinsic role in maintaining homeostasis of mature B cells and B-1 cells. FBW7 deletion led to an impairment of antibody response, and although germinal centre formation was not affected, antibody class-switch recombination and affinity maturation processes were defective. Likewise, memory immune response was severely impaired. Moreover, FBW7 ablation ameliorated the pathogenesis of an autoimmune disease model, collagen-induced arthritis, by reducing the production of anti-collagen II autoantibodies. Taken together, these data suggest that FBW7 may be an attractive target for developing new therapeutics for the treatment of autoimmune diseases.

Regulation of humoral immune response by HIF-1α-dependent metabolic reprogramming of the germinal center reaction.

Hypoxia-inducible factor-1α (HIF-1α) has been implicated in the regulation of many genes responsible for aerobic glycolysis; however, the role of HIF-1α in B-cell metabolism has not been well defined. Here, we analyzed patterns of gene expression and oxygen consumption rates in B-cell subpopulations from humans and mice and described a model of HIF-1α-mediated B-cell metabolic reprogramming during the germinal center (GC) reaction. Importantly, we found that HIF-1α was highly expressed in GC B-cells, and HIF-1α deficiency in B-cells impaired a functional GC reaction, resulting in defective class-switch recombination and generation of high-affinity plasma cells. These results identified an important role of HIF-1α in regulating humoral immunity through metabolic reprogramming during the GC response. This newly discovered metabolic character of GC B-cells will advance our understanding of GC biology and B-cell lymphomagenesis.

Lipid Receptor G2A-Mediated Signal Pathway Plays a Critical Role in Inflammatory Response by Promoting Classical Macrophage Activation.

Macrophage polarization is a dynamic and integral process in tissue inflammation and remodeling. In this study, we describe that lipoprotein-associated phospholipase A2 (Lp-PLA2) plays an important role in controlling inflammatory macrophage (M1) polarization in rodent experimental autoimmune encephalomyelitis (EAE) and in monocytes from multiple sclerosis (MS) patients. Specific inhibition of Lp-PLA2 led to an ameliorated EAE via markedly decreased inflammatory and demyelinating property of M1. The effects of Lp-PLA2 on M1 function were mediated by lysophosphatidylcholine, a bioactive product of oxidized lipids hydrolyzed by Lp-PLA2 through JAK2-independent activation of STAT5 and upregulation of IRF5. This process was directed by the G2A receptor, which was only found in differentiated M1 or monocytes from MS patients. M1 polarization could be inhibited by a G2A neutralizing Ab, which led to an inhibited disease in rat EAE. In addition, G2A-deficient rats showed an ameliorated EAE and an inhibited autoimmune response. This study has revealed a mechanism by which lipid metabolites control macrophage activation and function, modification of which could lead to a new therapeutic approach for MS and other inflammatory disorders.

Docosahexaenoic acid ameliorates autoimmune inflammation by activating GPR120 signaling pathway in dendritic cells.

Although the phenomenon that omega-3 polyunsaturated fatty acids (n-3 PUFAs) shows to have a beneficial effect in patients suffering from multiple sclerosis (MS) and other autoimmune diseases has been empirically well-documented, the molecular mechanisms that underline the anti-inflammatory effects of n-3 PUFAs are yet to be understood. In experimental autoimmune encephalomyelitis (EAE), a model for MS, we show that one of the underlying mechanisms by which dietary docosahexaenoic acid (DHA) exerts its anti-inflammatory effect is regulating the functional activities of dendritic cells (DCs). In DHA-treated EAE mice, DCs acquire a regulatory phenotype characterized by low expression of co-stimulatory molecules, decreased production of pro-inflammatory cytokines, and enhanced capability of regulatory T-cell induction. The effect of DHA on DCs is mediated by the lipid-sensing receptor, G protein-coupled receptor 120 (GPR120). A GPR120-specific small-molecule agonist could ameliorate the autoimmune inflammation by regulating DCs, while silencing GPR120 in DCs strongly increased the immunogenicity of DCs. Stimulation of GPR120 induces suppressor of cytokine signaling 3 (SOCS3) expression and down-regulates signal transducer and activator of transcription 3 (STAT3) phosphorylation, explaining the molecular mechanism for regulatory DC induction.

A chromosome-level genome of the mud crab (Scylla paramamosain estampador) provides insights into the evolution of chemical and light perception in this crustacean.

Mud crabs, found throughout the Indo-Pacific region, are coastal species that are important fisheries resources in many tropical and subtropical Asian countries. Here, we present a chromosome-level genome assembly of a mud crab (Scylla paramamosain). The genome is 1.55 Gb (contig N50 191 kb) in length and encodes 17,821 proteins. The heterozygosity of the assembled genome was estimated to be 0.47%. Effective population size analysis suggested that an initial large population size of this species was maintained until 200 thousand years ago. The contraction of cuticle protein and opsin genes compared with Litopenaeus vannamei is assumed to be correlated with shell hardness and light perception ability, respectively. Furthermore, the analysis of three chemoreceptor gene families, the odorant receptor (OR), gustatory receptor (GR) and ionotropic receptor (IR) families, suggested that the mud crab has no OR genes and shows a contraction of GR genes and expansion of IR genes. The numbers of the three gene families were similar to those in three other decapods but different from those in two nondecapods and insects. In addition, IRs were more diversified in decapods than in nondecapod crustaceans, and most of the expanded IRs in the mud crab genome were clustered with the antennal IR clades. These findings suggested that IRs might exhibit more diverse functions in decapods than in nondecapods, which may compensate for the smaller number of GR genes. Decoding the S. paramamosain genome not only provides insight into the genetic changes underpinning ecological traits but also provides valuable information for improving the breeding and aquaculture of this species.

High resolution laser Thomson scattering system with automatic data analysis software platform for diagnosis of the low-temperature plasmas.

Accurate measurements of electron temperature (Te) and electron density (ne) are important for understanding the properties of plasma, especially for the low-temperature plasma dominated by the free electrons. In this work, a high resolution laser Thomson scattering (LTS) diagnosis system with a software platform for processing data is established to accurately measure the Te and ne in low-temperature plasmas. In this system, a homemade Triple Grating Spectrometer (TGS) is elaborated to suppress the intense stray light and Rayleigh scattering light. The TGS with high etendue (with an f-number of f/3.3) and high spectral resolution (0.07 nm in full-width-half-maximum at 532 nm) allows the LTS system to measure the plasma with the ne and Te as low as 1.0 × 1017 m-3 and 0.1 eV, respectively. Besides the development of the experimental setup, a software platform is specially designed to automatically process the complicated LTS spectra to determine ne and Te in real time during the acquirement of data. The error analyses indicate that the uncertainty of ne is around 12% and the uncertainty of Te is about 10% when Te > 0.2 eV. Using the LTS system, we have successfully measured ne and Te in the cascaded arc plasma, nanosecond pulse discharge plasma, and inductively coupled plasma. The results demonstrate that the LTS system has significant potential for the diagnosis of various low temperature plasmas.

Genetic variation and forensic efficiency of 30 indels for three ethnic groups in Guangxi: relationships with other populations.

AIM: In this study, we used a series of diallelic genetic marker insertion/deletion polymorphism (indel) to investigate three populations of Yao, Kelao, and Zhuang groups in the Guangxi region of China and to evaluate their efficiency in forensic application. RESULT: No deviations for all 30 loci were observed from the Hardy-Weinberg equilibrium after Bonferroni correction (p > 0.05/30 = 0.0017). The allele frequencies of the short allele (DIP-) for the above three populations were in the range of 0.0520-0.9480, 0.0950-0.8780, and 0.0850-0.915, respectively. The observed heterozygosity of the 30 loci for the three populations was in the ranges 0.0802-0.5802, 0.1908-0.6053, and 0.1400-0.5600, respectively. The cumulative power of exclusion and combined discrimination power for Yao, Kelao, and Zhuang groups were (0.9843 and 0.9999999999433), (0.9972 and 0.9999999999184), and (0.9845 and 0.9999999999608), respectively. The DA distance, principal component analysis, and cluster analysis indicated a clear regional distribution. In addition, Zhuang groups had close genetic relationships with the Yao and Kelao populations in the Guangxi region. CONCLUSION: This study indicated that the 30 loci were qualified for personal identification; moreover, they could be used as complementary genetic markers for paternity testing in forensic cases for the studied populations.

Genetic composition of Teochew population in Chaozhou revealed by 19 STR loci: population polymorphism and interpopulation comparisons.

Aim: To investigate the genetic polymorphisms of 19 STR loci in the Teochew population of Chaozhou. Subjects and methods: Nineteen STR loci of 631 unrelated Teochew people in Chaozhou were detected by using the AGCU Expressmarker 20 kit. Results: The allele frequency ranged from 0.0008-0.5577. The combined power of discrimination and combined probability of excluding paternity is over 0.999,999,999,999,999,999,999,999,999,999 and 0.999,999,992,549,546, respectively. The principal component analysis based on the 11 shared STR loci showed the Teochew population was most genetically related to the Fujian Han population. Conclusion: The results demonstrate the set of 19 autosomal STRs could provide robust genetics information for individual identification, paternity testing and human genetics research in the Teochew population.

Developmental validation of a novel six-dye typing system with 47 A-InDels and 2 Y-InDels.

In this study, a multiplex amplification system including 47 autosomal InDels, 2 Y-chromosome InDels, and the sex-determining marker (Amelogenin) was developed with six fluorescent dyes labeling. These InDels were selected from the previous study based on a series of criteria (0.3 < MAF < 0.5, HET > 0.4, etc). The system was designated the AGCU InDel 50 kit and was validated in a series of studies, including a degradation study; tests for sensitivity, species specificity, reproducibility, stability, applicability to case samples, balance of peak height, and PCR conditions; and a population study. The results showed that AGCU InDel 50 kit was quite sensitive, specific, stable in several PCR conditions or exposure to PCR inhibitors, especially against degradation. 74 case samples and 50 paternity cases with STR mutation events were tested using PowerPlex® 21 System, AGCU InDel 50 kit, and Investigator DIPplex kit, and the results showed that the ratio of loci detected with the developed kit were close to Investigator@ DIPplex kit, but considerably higher than PowerPlex® 21 System for case samples containing low amounts of degraded DNA. As for 50 paternity cases, no mutation was observed in any InDels locus, and the CPIs based on 47 autosomal InDels contained in the AGCU InDel 50 kit were all higher than those based on 30 InDels contained in Investigator® DIPplex kit, except 3 cases. In the population study, 203 unrelated individuals from the Guangdong Han population were detected using the AGCU InDel 50 kit, and the values of combined power of discrimination and combined power of exclusion were 0.999 999 999 999 999 and 0.9997, respectively. Thus, AGCU InDel 50 kit is suitable for individual identification and as a supplemental tool for paternity testing. It is reproducible, accurate and robust for forensic applications and human genetic studies.

Identification of extra alleles in DYS385a/b multi-allelic patterns.

Y chromosome short tandem repeats (Y-STRs) are widely used in forensic DNA analysis. Y-STRs usually genotyped as a monoallelic pattern, or a diallelic pattern at some double-copy loci. Abnormal genotypes with three or four alleles have been reported at DYS385a/b, which is a double-copy locus. However, multi-allelic patterns with more than four alleles have not been reported at this locus. In this study, abnormal five-allelic patterns at DYS385a/b locus were observed in 2 out of 7760 unrelated males from Henan Province, detected by the HG19+14Y fluorescence detection kit; and abnormal six-allelic pattern was observed in 1 out of 4920 unrelated males from Guangdong Province, detected by the AGCU Data Y30 fluorescence kit. The genotypes of these abnormal samples were confirmed again by the Yfiler® Plus and PowerPlex® Y23 kits. In order to understand the genomic structure of the extra alleles at this locus, forward and reverse primers were designed to sequence the three samples. The results showed that the five-banded and six-banded pattern genotype did exist at DYS385a/b locus. These data enriched the knowledge about multi-allelic patterns at DYS385a/b locus, which allowed the use of DYS385a/b profile in forensic analysis, even with multi-allelic patterns.

Genetic polymorphisms of 20 STR loci in a Chaoshan group from Jieyang, China.

We obtained allelic frequencies and forensic parameters of 20 short tandem repeat (STR) loci (D3S1358, FGA, D5S818, D7S820, CSF1PO, D16S539, D19S433, vWA, D8S1179, D18S51, D13S317, TPOX, TH01, D2S1338, D12S391, D1S1656, D21S11, D6S1043, Penta D, Penta E) from 529 unrelated individuals in Jieyang Han population using PowerPlex® 21 (Promega, Madison, Wi, USA). The relationship between the Jieyang Han group and other Han populations was studied and the results showed that the Jieyang Han population had the closest genetic relationship with the Fujian Han population.

De novo sequencing of the Antarctic krill (Euphausia superba) transcriptome to identify functional genes and molecular markers.

To provide massive genetic resources for the Antarctic krill (Euphausia superba), we sequenced and analysed the transcriptome by using high-throughput Illumina paired-end sequencing technology. A total of 77.1 million clean reads representing ~11.0Gb data were generated. The average length of these reads was 142 bp. De novo assembly yielded 125,211 transcripts with a N50 of 690 bp. Further analysis produced 106,250 unigenes, of which 31,683 were annotated based on protein homology searches against protein databases. Gene ontology analysis showed that ion binding, organic substance, metabolic process, and cell part were the most abundantly used terms in molecular function, biological process and cellular component categories, respectively. In addition, 3067 unigenes were mapped onto 311 signal pathways by the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Finally, 15,224 simple sequence repeats were identified from 13,535 transcripts, and 103,593 single-nucleotide polymorphisms were found from 21.6% of total transcripts. These genetic resources obtained in this study forms a good foundation for investigating gene function, and evaluating population genetic diversity for this important Southern Ocean fisheries resource, E. superba.

Remote in situ laser-induced breakdown spectroscopic approach for diagnosis of the plasma facing components on experimental advanced superconducting tokamak.

The diagnosis of the fuel retention and impurity deposition on the plasma facing components (PFCs) is very important for monitoring plasma-wall interactions and improving the performance of long-pulse operation for tokamak devices. In this study, a remote in situ laser-induced breakdown spectroscopic (RIS-LIBS) system has been developed to be an effective and routine method for the diagnosis of the composition of the PFCs on Experimental Advanced Superconducting Tokamak (EAST). The RIS-LIBS system can be operated between EAST discharges via a remote network control system. This allows a flexible diagnosis for the PFCs at a specific EAST discharge operation or under planned plasma scenarios according to the experimental requirement. Measurements on the fuel retention and impurity deposition of the PFCs have been performed for the test of the RIS-LIBS system, and the depth resolution and the lateral resolution of the RIS-LIBS system have been achieved to be ∼100 nm and ∼3.0 mm, respectively. For the test of detectable elements, the fuel (deuterium) and impurities have been detected and identified clearly. In addition, the measurement of fuel abundance on the first wall as a function of the days of EAST deuterium plasma discharges has been carried out for the first time. These results well manifest a significant prospect of the RIS-LIBS for the diagnosis of the PFCs in the upcoming fusion devices like China Fusion Engineering Test Reactor (CFETR) and ITER.

A dynamic monitoring approach for the surface morphology evolution measurement of plasma facing components by means of speckle interferometry.

Plasma Facing Components (PFCs) in a magnetically confined fusion plasma device will be exposed to high heat load and particle fluxes, and it would cause PFCs' surface morphology to change due to material erosion and redeposition from plasma wall interactions. The state of PFCs' surface condition will seriously affect the performance of long-pulse or steady state plasma discharge in a tokamak; it will even constitute an enormous threat to the operation and the safety of fusion plasma devices. The PFCs' surface morphology evolution measurement could provide important information about PFCs' real-time status or damage situation and it would help to a better understanding of the plasma wall interaction process and mechanism. Meanwhile through monitoring the distribution of dust deposition in a tokamak and providing an upper limit on the amount of loose dust, the PFCs' surface morphology measurement could indirectly contribute to keep fusion operational limits and fusion device safety. Aiming at in situ dynamic monitoring PFCs' surface morphology evolution, a laboratory experimental platform DUT-SIEP (Dalian University of Technology-speckle interferometry experimental platform) based on the speckle interferometry technique has been constructed at Dalian University of Technology (DUT) in China. With directional specific designing and focusing on the real detection condition of EAST (Experimental Advanced Superconducting Tokamak), the DUT-SIEP could realize a variable measurement range, widely increased from 0.1 µm to 300 µm, with high spatial resolution (<1 mm) and ultra-high time resolution (<2 s for EAST measuring conditions). Three main components of the DUT-SIEP are all integrated and synchronized by a time schedule control and data acquisition terminal and coupled with a three-dimensional phase unwrapping algorithm, the surface morphology information of target samples can be obtained and reconstructed in real-time. A local surface morphology of the real divertor tiles adopted from EAST has been measured, and the feasibility and reliability of this new experimental platform have been demonstrated.

Forensic efficiency and genetic variation of 30 InDels in Vietnamese and Nigerian populations.

Insertion/deletion polymorphisms (InDels) are ubiquitous diallelic genetic markers that have drawn increasing attention from forensic researchers. Here, we investigated 30 InDel loci in Vietnamese and Nigerian populations and evaluated their usefulness in forensic genetics. The polymorphic information content of these populations ranged, respectively, from 0.164 to 0.375 and from 0.090 to 0.375 across loci. After Bonferroni correction, no significant deviation from Hardy-Weinberg equilibrium was found, except for HLD97 in the Nigerian population. The cumulative power of exclusion for all 30 loci in the Vietnamese and Nigerian populations was 0.9870 and 0.9676, respectively, indicating that this InDel set is not suitable for paternity testing in these populations, but could be included as a supplement. For the Vietnamese and the Nigerian populations, the mean observed heterozygosity was 0.5917 and 0.6268, and the combined discrimination power of the 30 loci was 0.9999999999767 and 0.9999999999603, respectively. These findings indicated that these InDels may be suitable for personal forensic identification in the studied populations. The results of DA distance, phylogenetic tree, principal component, and cluster analyses were consistent and indicated a clear pattern of regional distribution. Moreover, the Vietnamese population was shown to have close genetic relationships with the Guangdong Han and Shanghai Han populations.

High sensitive and high temporal and spatial resolved image of reactive species in atmospheric pressure surface discharge reactor by laser induced fluorescence.

The current paucity of spatial and temporal characterization of reactive oxygen and nitrogen species (RONS) concentration has been a major hurdle to the advancement and clinical translation of low temperature atmospheric plasmas. In this study, an advanced laser induced fluorescence (LIF) system has been developed to be an effective antibacterial surface discharge reactor for the diagnosis of RONS, where the highest spatial and temporal resolution of the LIF system has been achieved to ∼100 µm scale and ∼20 ns scale, respectively. Measurements on an oxidative OH radical have been carried out as typical RONS for the benchmark of the whole LIF system, where absolute number density calibration has been performed on the basis of the laser Rayleigh scattering method. Requirements for pixel resolved spatial distribution and outer plasma region detection become challenging tasks due to the low RONS concentration (∼ppb level) and strong interference, especially the discharge induced emission and pulsed laser induced stray light. In order to design the highly sensitive LIF system, a self-developed fluorescence telescope, the optimization of high precision synchronization among a tunable pulsed laser, a surface discharge generator, intensified Charge Coupled Device (iCCD) camera, and an oscilloscope have been performed. Moreover, an image BOXCAR approach has been developed to remarkably improve the sensitivity of the whole LIF system by optimizing spatial and temporal gating functions via both hardware and software, which has been integrated into our automatic control and data acquisition system on the LabVIEW platform. In addition, a reciprocation averaging measurement has been applied to verify the accuracy of the whole LIF detecting system, indicating the relative standard deviation of ∼3%.

Complete mitochondrial genome and phylogenetic position of Chaenodraco wilsoni.

Chaenodraco wilsoni, a species of Channichthyidae, inhabits in southern ocean. The total length of complete mitochondrial genome of C. wilsoni is 17,432 bp, which encoded 37 genes. Similar to most Antarctic fishes, the ND6/tRNA (glu) translocation and an additional non-coding region linked with ND6 have also occurred in C. wilsoni. The ML tree supports that C. wilsoni has closer relationship with Chionodraco species. Our research will provide more molecular biology information about C. wilsoni and deepen the understanding of Antarctic fishes.

Complete mitochondrial genome of Antarctic krill Euphausia superba (Eucarida: Euphausiacea).

Antarctic krill Euphausia superba is a very important species in Antarctic ecosystem. The mitochondrial genome of E. superba was completed with 16,591 bp in length, gene arrangement and order was the same as previous studies. The overall A + T content is 68.91% and the control region A + T content is 78.17%. Alignment with other two partial mitochondrial genome revealed that ND2 region possessed many unusual variation sites. The phylogenetic tree showed that E. superba clustered with other two species of Euphausiacea. This study filled in the gap of the krill mitogenome and suggested putative markers for population study.

The first complete mitochondrial genome sequence of Uraspis secunda (Perciformes: Carangidae) and its phylogenetic relationship.

In this study, the first complete mitochondrial genome sequence of Uraspis secunda was determined by using Illumina and Sanger sequencing technology. This genome was 16 554 bp long, consisting of 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and a putative control region. Twenty-eight genes were located on heavy strand, while nine genes were located on light strand. Totally two types of start codons and four kinds of stop codons were observed from 13 protein-coding genes. The overall base composition of this genome was 28.17% for A, 16.23% for G, 25.81% for T, and 29.78% for C, with a slightly higher A + T content of 53.99%. The highest A + T content (63.18%) was found in the control region of the mitochondrial genome. A phylogenetic tree was constructed based on 13 concatenated protein-coding genes, which indicated that U. secunda was closest to Parastromateus niger, and these two fish species clustered a 1. monophyletic group with Alectis ciliaris, Carangoides malabaricus and C. armatus. This study will be helpful for researches on molecular phylogeny and genetic diversity of U. secunda and the related fish species.