235 results for qubit oscillator frequency
Applying a field spectroscopy technique for assessing successional trends of biological soil crusts in a semi-arid environment
Contributors: E. Zaady, A. Karnieli, M. Shachak
... We studied the successional stages of biological soil crusts (BSCs) by using in-situ spectroscopic techniques during 6 years of recovery following scraping-sterilization and scraping-crumbling disturbances on north- and south-facing slopes and in plots with and without overland water runoff barriers. Two spectral indices, the Brightness Index (BI) and the Normalized Difference Vegetation Index (NDVI), were used as indicators for evaluating BSC succession, with special attention to differences between the north- and the south-facing slopes. We found that BSC succession could be expressed as linear regressions of the above-mentioned indicators during the experimental years for the different treatments. Both indicators were found to be significantly different in each of the experimental years: BI values decreased while NDVI values increased for each of the three treatments. Thus, the BI can serve as a good indicator during the early years following disturbance while the NDVI can be useful after crusts have become established. We conclude that spectral reflectance measurements of BSCs can be a useful monitoring technique for studying the regeneration of the soil surface without direct disturbance.
CRACC-targeting Fc-fusion protein induces activation of NK cells and DCs and improves T cell immune responses to antigenic targets
Contributors: Yasser A. Aldhamen, David P.W. Rastall, Weimin Chen, Sergey S. Seregin, Cristiane Pereira-Hicks, Sarah Godbehere, Norbert E. Kaminski, Andrea Amalfitano
Development of CRACC-Fc fusion protein. (A) Plasmid encoding CRACC-ECD/mIgG1-Fc was transiently transfected into 100ml suspension HEK 293-6E cell culture. CRACC-Fc protein was captured from the cell culture supernatant by HiTrapTM Mabselect 5ml column and followed by buffer exchange and SDS-PAGE and Western blot analysis. Lane M: protein marker. Lane R: reducing conditions. Lane N: non-reducing conditions. Lane P: mouse IgG1, Kappa (B and C) RAW264.7 cells (300,000cells/well) were plated in quadruplicate in 24-well plate and then stimulated for 24h with LPS (1μg/ml). After 24h, cells were harvested and incubated with escalating doses of CRACC-Fc fusion protein for 2h. After 2h, cells were then washed and stained with APC-conjugated anti-CRACC antibody and CRACC expression was evaluated by flow cytometry. (B) Representative flow cytometry figures for CRACC expression on LPS-stimulated RAW264.7 cells in the presence or absence of CRACC-Fc fusion protein. (C) Frequency of CRACC expression on LPS-stimulated RAW264.7 cells in the presence of escalating doses of CRACC-Fc fusion protein. (D) Immunoprecipitation analysis for EAT-2 expression in RAW264.7 cells following LPS stimulation in the presence or absence of CRACC-Fc (10μg/ml). (E) Quantification of EAT-2 protein levels after normalization to the 55kDa loading protein. Densitometric analysis was done using Image J software. Cells were plated in quadruplicate. The bars represent mean±SD. ...Frequency of T and B lymphocytes following rAd5-Null and CRACC-Fc co-administration. (A) Flow cytometry analysis for the percentages of CD8+ CD3+ T cells (A), CD8− CD3+ T cells (B), and CD19+ CD3− B cells (C) 12 days following rAd5-Null and either CRACC-Fc or control IgG co-administration. Statistical analysis was completed using a Student's t-test, p<0.05 was deemed as a statistically significant difference. ...CRACC-Fc fusion protein has an innate immunostimulatory activity. Male C57BL/6 mice (n=5) were i.p. injected with a mixture of rAd5-Null (2×1010vps/mouse) and 200μg of CRACC-Fc fusion protein [rAd5-Null+ CRACC-Fc) or control IgG [Ad-Null+ Control IgG]. After 12h, mice were sacrificed and splenocytes were prepared. (A) Frequency of IFNγ expressing NK cells (CD3-NK1.1+) derived from Ad-Null+ CRACC-Fc or Ad-Null+ Control IgG co-injected mice. (B and C) DCs (CD11c+, CD11b+, F4/80−) maturation was evaluated by assessing the expression levels of CD80, CD86, and I-A/I-E molecules. Expression of CD80 (B), CD86 (C), and I-A/I-E (D) on CD11c+, CD11b+, F4/80− DCs is shown. Data are presented as mean±SD. Statistical analysis was completed using a one-way analysis of variance (ANOVA) with a Student–Newman–Keuls post hoc test, p<0.05 was deemed as a statistically significant difference. *** denotes p<0.001, statistically different from naive animals. ...Cytokines secretion profiles of Gag-specific CD8+ T cells derived from rAd5-HIV/Gag and CRACC-Fc co-vaccination. Splenocytes (2×106cells/well) derived from naives (n=4) or vaccinated mice (n=7) were stimulated with the HIV-Gag immunogenic peptide, AMQMLKETI and FACS ICS analysis was performed. The Cytokines secretion profiles of HIV-Gag-specific CD8+ T cells elicited by the rAd5-HIV/Gag and CRACC-Fc co-vaccination regimen were determined by multiparameter intercellular staining assay. Gate were set based on negative control (naïve) and placed consistently across samples. The total frequency of splenic CD8+ T cells derived from naïve or vaccinated mice expressing IFNγ (A), TNFα (B), IL-2 (C), IFNγ/TNFα (D), IFNγ/IL-2 (E), or TNFα/IL-2 (F). Data are presented as mean±SD. Statistical analysis was completed using a one-way analysis of variance (ANOVA) with a Student–Newman–Keuls post hoc test, p<0.05 was deemed as a statistically significant difference. **, *** denotes p<0.01, p<0.001 statistically different from naive animals. Data are representative of three independent experiments with similar results. ...CRACC-Fc co-vaccination alters the elicited CD8+ T cells toward TEM phenotype. Phenotypic analysis of HIV-Gag-specific CD8+ T cells elicited after rAd5-HIV/Gag and CRACC-Fc co-vaccination at 42dpi. Multiparameter Gag-tetramer binding assays were performed in PBMCs (A) or splenocytes (B) of naives (n=4) or vaccinated mice (n=6) to enumerate the frequency of Gag-specific tetramer positive effector memory (CD62Llow CD127high) CD8+ T cells. Data are presented as mean±SD. Statistical analysis was completed using a one-way analysis of variance (ANOVA) with a Student–Newman–Keuls post hoc test, p<0.05 was deemed as a statistically significant difference. **, *** denotes p<0.01, p<0.001 statistically different from naive animals. Data are representative of three independent experiments with similar results. ...Increased breadth of Gag-specific CD8+ T cell responses following rAd5-HIV/Gag and CRACC-Fc co-vaccination. Splenocytes (2×106cells/well) derived from naives (n=4) or vaccinated mice (n=7) were stimulated with Gag-peptide pool# 17 (identified in Fig. 4C) and FACS ICS analysis for IFNγ-, TNFα- and IL-2-expressing CD8+ T cells was performed. The total frequency of splenic CD8+ T cells expressing IFNγ (A), TNFα (B), IL-2 (C), IFNγ/TNFα (D), IFNγ/IL-2 (E), or TNFα/IL-2 (F) are shown. Data are presented as mean±SD. Statistical analysis was completed using a one-way analysis of variance (ANOVA) with a Student–Newman–Keuls post-hoc test, p<0.05 was deemed as a statistically significant difference. **, *** denotes p<0.01, p<0.001 statistically different from naive animals. Data are representative of two independent experiments with similar results. ...CRACC-Fc fusion protein has an innate immunostimulatory activity. Male C57BL/6 mice (n=5) were i.p. injected with a mixture of rAd5-Null (2×1010vps/mouse) and 200μg of CRACC-Fc fusion protein [rAd5-Null+ CRACC-Fc) or control IgG [Ad-Null+ Control IgG]. After 12h, mice were sacrificed and splenocytes were prepared. (A) Frequency of IFNγ expressing NK cells (CD3-NK1.1+) derived from Ad-Null+ CRACC-Fc or Ad-Null+ Control IgG co-injected mice. (B and C) DCs (CD11c+, CD11b+, F4/80−) maturation was evaluated by assessing the expression levels of CD80, CD86, and I-A/I-E molecules. Expression of CD80 (B), CD86 (C), and I-A/I-E (...Increased breadth of Gag-specific CD8+ T cell responses following rAd5-HIV/Gag and CRACC-Fc co-vaccination. Splenocytes (2×106cells/well) derived from naives (n=4) or vaccinated mice (n=7) were stimulated with Gag-peptide pool# 17 (identified in Fig. 4C) and FACS ICS analysis for IFNγ-, TNFα- and IL-2-expressing CD8+ T cells was performed. The total frequency of splenic CD8+ T cells expressing IFNγ (A), TNFα (B), IL-2 (C), IFNγ/TNFα (D), IFNγ/IL-2 (E), or TNFα/IL-2 (F) are shown. Data are presented as mean±SD. Statistical analysis was completed using a one-way analysis of variance (ANOVA) with a Student–Newman–Keuls post-hoc test, psimilar results. ... The CD2-like receptor activating cytotoxic cell (CRACC) receptor is a member of the SLAM family of receptors that are found on several types of immune cells. We previously demonstrated that increasing the abundance of the adaptor protein EAT-2 during vaccination enhanced innate and adaptive immune responses to vaccine antigens. Engagement of the CRACC receptor in the presence of the EAT-2 adaptor generally results in immune cell activation, while activating CRACC signaling in cells that lack EAT-2 adaptor inhibits their effector and regulatory functions. As EAT-2 is the only SAP adaptor that interacts with the CRACC receptor, we hypothesized that technologies that specifically modulate CRACC signaling during vaccination may also improve antigen specific adaptive immune responses. To test this hypothesis, we constructed a CRACC-targeting Fc fusion protein and included it in vaccination attempts. Indeed, mice co-vaccinated with the CRACC-Fc fusion protein and an adenovirus vaccine expressing the HIV-Gag protein had improved Gag-specific T cell responses, as compared to control mice. These responses are characterized by increased numbers of Gag-specific tetramer+ CD8+ T cells and increases in production of IFNγ, TNFα, and IL2, by Gag-specific CD8+ T cells. Moreover, our results revealed that use of the CRACC-Fc fusion protein enhances vaccine-elicited innate immune responses, as characterized by increased dendritic cells (DCs) maturation and IFNγ production from NK cells. This study highlights the importance of CRACC signaling during the induction of an immune response generally, and during vaccinations specifically, and also lends insight into the mechanisms underlying our prior results noting EAT-2-dependent improvements in vaccine efficacy.
Contributors: Morteza Fattahi, Richard T. Walker
... Iran is one of the world's most tectonically active regions, yet dating past earthquakes for neotectonic studies has been limited. One of the main reasons for this is that organic material suitable for radiocarbon dating of deformed sediments is rare. We investigate the use of infrared stimulated luminescence (IRSL) from coarse-grained feldspars to date colluvial deposits associated with the Sabzevar thrust fault in northeastern Iran. The single-aliquot regenerative (SAR) dose measurement procedure was used for this study. The current study investigates monitoring and correcting for sensitivity changes, recovering a known laboratory dose and equivalent dose estimation using three SAR IRSL methods. It is shown that SAR has recovered a given laboratory dose using a range of preheat temperatures but De determination of natural samples requires its own preheat plateaus for two of these SAR methods. The SAR IRSL method provided an age of 1.7±0.3ka for colluvium, predating the last earthquake event on the Sabzevar fault. This result suggests that this fault is likely to be responsible for an earthquake that destroyed Sabzevar city in AD 1052.
Original article - Immunomodulation and signaling mechanism of Lactobacillus rhamnosus GG and its components on porcine intestinal epithelial cells stimulated by lipopolysaccharide
Contributors: Kan Gao, Chong Wang, Li Liu, Xiaoxiao Dou, Jianxin Liu, Lijuan Yuan, Wenming Zhang, Haifeng Wang
... This study aimed to evaluate the immunomodulatory effects and signaling mechanisms of Lactobacillus rhamnosus GG (LGG) and its components [surface-layer protein (SLP), DNA, exopolysaccharides, and CpG oligodeoxynucleotides] on lipopolysaccharide (LPS)-stimulated porcine intestinal epithelial cell (IEC) IPEC-J2.
Contributors: Masashi Terao, Shirin Akter, Md. Golam Yasin, Ryo Nakao, Hirotomo Kato, Mohammad Zahangir Alam, Ken Katakura
... Babesia gibsoni is a tick-borne hemoprotozoan parasite of dogs that often causes fever and hemolytic illness. Detection of B. gibsoni has been predominantly reported in Asian countries, including Japan, Korea, Taiwan, Malaysia, Bangladesh and India. The present study shows the first molecular characterization of B. gibsoni detected from dogs in Bangladesh. Blood samples were collected on FTA® Elute cards from 50 stray dogs in Mymensingh District in Bangladesh. DNA eluted from the cards was subjected to nested PCR for the 18S rRNA gene of Babesia species. Approximately 800bp PCR products were detected in 15 of 50 dogs (30%). Based on restriction fragment length polymorphism (RFLP) and direct sequencing of the PCR products, all parasite isolates were identified as B. gibsoni. Furthermore, the BgTRAP (B. gibsoni thrombospondin-related adhesive protein) gene fragments were detected in 13 of 15 18S rRNA gene PCR positive blood samples. Phylogenetic analysis of the BgTRAP gene revealed that B. gibsoni parasites in Bangladesh formed a cluster, which was genetically different from other Asian B. gibsoni isolates. In addition, tandem repeat analysis of the BgTRAP gene clearly showed considerable genetic variation among Bangladeshi isolates. These results suggested that B. gibsoni parasites in a different genetic clade are endemic in dogs in Bangladesh. Further studies are required to elucidate the origin, distribution, vector and pathogenesis of B. gibsoni parasites circulating in dogs in Bangladesh.
Genetic and evolutionary analysis of cell-fusing agent virus based on Thai strains isolated in 2008 and 2012
Contributors: Atsushi Yamanaka, Supatra Thongrungkiat, Pongrama Ramasoota, Eiji Konishi
... Increasing attention is being devoted to ecological and evolutionary relationships between insect-specific flaviviruses and globally important human-pathogenic flaviviruses such as dengue viruses. One such insect flavivirus, cell-fusing agent virus (CFAV), remains poorly investigated. In this study, we isolated 13 and 16 CFAV strains from Aedes aegypti mosquitoes collected in Thailand in 2008 and 2012, respectively, and performed genetic and evolutionary analyses based on gene regions encoding the envelope protein (E) and nonstructural proteins 3 (NS3) and 5 (NS5). Consistent with previously reported CFAV strains, E, NS3 and NS5 regions comprised 1,290, 1,761 and 2,664 nucleotides, respectively. Nucleotide and amino acid identities of these three regions were >98% among the 29 isolates, and approximately 95–96% and 96–99%, respectively, between the isolates and previously reported CFAV strains. When amino acid sequences from representative strains of six insect-specific and seven mosquito-borne flaviviruses were compared, average identities of 14.9%, 31.8% and 44.3% were calculated for E, NS3 and NS5 regions, respectively. Phylogenetic analysis based on nucleotide and amino acid data indicated that the Thai CFAV isolates of the current study were distinct from previously reported CFAV strains from Indonesia and Puerto Rico. Analysis of each gene region consistently uncovered a clade made up of nearly the same subset of Thai CFAV isolates; this result, and the isolation of CFAV from mosquitoes reared from larvae, suggest that the virus is maintained by vertical transmission and conserved in a particular environment without considerable evolutionary alteration. The most recent common ancestor of the Thai CFAV isolates in this study was dated to 11–27years ago, and is estimated to have diverged 46–86years ago from previously reported CFAV strains. Superinfection with CFAV of Aedes mosquitoes carrying dengue viruses present in Thailand for over 50years has most likely taken place.
Strong modulation of ectopic focus as a mechanism of repetitive interpolated ventricular bigeminy with heart rate doubling
Contributors: Kan Takayanagi, Shiro Nakahara, Noritaka Toratani, Ryuji Chida, Sayuki Kobayashi, Yoshihiko Sakai, Akihiro Takeuchi, Noriaki Ikeda
... Repetitive interpolated ventricular bigeminy (RIVB) can introduce a doubling of the ventricular rate.
Organ-specific proteomics analysis for identification of response mechanism in soybean seedlings under flooding stress
Contributors: Amana Khatoon, Shafiq Rehman, Susumu Hiraga, Takahiro Makino, Setsuko Komatsu
... Flooding is one of the severe environmental factors which impair growth and yield in soybean plant. To investigate the organ specific response mechanism of soybean under flooding stress, changes in protein species were analyzed using a proteomics approach. Two-day-old soybeans were subjected to flooding for 5days. Proteins were extracted from root, hypocotyl and leaf, and separated by two-dimensional polyacrylamide gel electrophoresis. In root, hypocotyl and leaf, 51, 66 and 51 protein species were significantly changed, respectively, under flooding stress. In root, metabolism related proteins were increased; however these proteins were decreased in hypocotyl and leaf. In all 3 organs, cytoplasm localized proteins were decreased, and leaf chloroplastic proteins were also decreased. Isoflavone reductase was commonly decreased at protein level in all 3 organs; however, mRNA of isoflavone reductase gene was up-regulated in leaf under flooding stress. Biophoton emission was increased in all 3 organs under flooding stress. The up-regulation of isoflavone reductase gene at transcript level; while decreased abundance at protein level indicated that flooding stress affected the mRNA translation to proteins. These results suggest that concurrence in expression of isoflavone reductase gene at mRNA and protein level along with imbalance in other disease/defense and metabolism related proteins might lead to impaired growth of root, hypocotyl and leaf of soybean seedlings under flooding stress.
Sperm phosphoproteome profiling by ultra performance liquid chromatography followed by data independent analysis (LC–MSE) reveals altered proteomic signatures in asthenozoospermia
Contributors: Priyanka P. Parte, Parimala Rao, Shweta Redij, Vivian Lobo, Serena J. D'Souza, Rahul Gajbhiye, Vijay Kulkarni
... Sperm motility is an important prerequisite for successful fertilization and is regulated by cyclic AMP activated protein kinase A which phosphorylates flagella proteins like axonemal dynein and initiates motility. Increase in calcium influx reverses this process by dephosphorylation that is mediated by calcineurin. Analyzing the phosphoenriched fractions of spermatozoa lysates from eight normozoospermic-, and asthenozoospermic-samples, respectively, by Nano UPLC–MSE, the present study investigates the phosphoproteins involved in sperm motility in an attempt to identify the key pathways regulating sperm motility and likely to be altered in spermatozoa of asthenozoospermic individuals. 66 phosphoproteins were differentially regulated in asthenozoospermia. The deregulated proteins comprised predominantly the HSPs, cytoskeletal proteins, proteins associated with the fibrous sheath, and those associated with energy metabolism. EM analysis of these spermatozoa demonstrated significant defects in mitochondria, and fibrous sheath and these defects could be correlated with the altered proteome. Pathway analysis revealed that carbohydrate and energy metabolism, cAMP mediated PKA signaling, PI3K/AKT signaling and pathway regulating actin based motility by Rho were significantly altered indicating that motility in spermatozoa is regulated through the concerted effort of these pathways. The data identified signature molecules that have the potential as biomarkers for diagnosing etiology of asthenozoospermia.
Contributors: Christin F. Down, Julie Millour, Eric W.-F. Lam, Roger J. Watson
... The promoters of genes which regulate entry into and progress through mitosis are typically induced maximally in G2 by transcription factors that include B-Myb and FoxM1. As FoxM1 gene transcription is a target of B-Myb, we investigated in this study how these transcription factors functionally interact to regulate these G2/M genes. Using a 3T3 cell line containing floxed B-myb alleles (B-mybF/F) that could be conditionally deleted by Cre recombinase, we confirmed that B-myb knockout caused both decreased mRNA expression of several G2/M genes, including FoxM1, and delayed entry into mitosis. Although FoxM1 protein expression was actually unaffected by B-myb knockout when quiescent B-mybF/F 3T3 cells re-entered the cell cycle upon serum-stimulation, chromatin immunoprecipitation revealed that FoxM1 binding to G2/M promoters was substantially reduced. FoxM1 transcriptional activity requires sequential phosphorylation by Cyclin-dependent kinases and Plk1, which are B-Myb target genes, and we found that phosphorylation at Plk1-specific sites was somewhat reduced upon B-myb knockout. Neither this effect nor nuclear accumulation of FoxM1, which was unaffected by B-myb knockout, was sufficient to account for the dependence on B-Myb for FoxM1 promoter binding, however. More significantly, assays using paired Birc5 (survivin) promoter-luciferase reporters with either wild-type or mutated Myb binding sites showed that FoxM1 was unable to bind and activate the promoter in the absence of B-Myb binding. Our data suggest that B-Myb is required as a pioneer factor to enable FoxM1 binding to G2/M gene promoters and explains how these transcription factors may collaborate to induce mitosis.