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Microarray profiling and functional analysis of differentially expressed plasma exosomal circular RNAs in Graves' disease
Ying Sun1 na1,
Wei Wang1 na1,
Yuxiao Tang1 na1,
Daping Wang1 na1,
Liang Li2,
Min Na3,
Guantong Jiang4,
Qian Li5,
Shulin Chen1 &
Jin Zhou ORCID: orcid.org/0000-0002-7482-40591 na1
Biological Research volume 53, Article number: 32 (2020) Cite this article
Circulating RNA (circRNA) regulates various bioactivities in cells. A better understanding of the exosomal circRNA can provide novel insights into the pathogenesis and treatment of Graves' disease (GD). We aimed to profile the differentially expressed circRNAs (DEcRs) in plasma exosomes of patients with GD and speculate and probe the functions of the DEcR by comprehensive bioinformatics analyses.
Serum exosomes were isolated from five primary GD patients and five healthy controls via ultracentrifugation. After verification with transmission electron microscopy, exosome samples were subjected to microarray profiling using human circRNA microarrays. Two up-regulated and two down-regulated DEcRs were selected for validation in plasma exosomes from 20 GD and 20 healthy control participants using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR). The circRNA/microRNA/mRNA interaction network was then assembled and the analysis of the Gene Ontology and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways was utilized to predict the potential functions of the DEcR associated genes.
There were 15 DEcRs revealed in primary GD cases. The intronic circRNA hsa_circRNA_000102 was confirmed as an up-regulated component in plasma exosomes from patients with GD. The circRNA/microRNA/mRNA interaction network unveiled the most potential targeting microRNAs of hsa_circRNA_000102 and its associated genes. The functional analyses predicted involvement of hsa_circRNA_000102 associated genes in pathways of immune system activation, such as viral infection and interferon-beta signaling.
hsa_circRNA_000102 is a differentially up-regulated plasma exosomal circRNA in patients with GD. Our study highlights multiple pathways, particularly virus infection and interferon-beta signaling, for mediating immune activation in Graves' disease.
Graves' disease (GD) is an autoimmune disorder characterized by typical clinical manifestations with unique association of diffuse goiter, ophthalmopathy, and pretibial myxedema [1]. GD is the most common cause of hyperthyroidism, identified in patients spanning the globe [2]. Timely diagnosis and appropriate treatments are critical for the prognosis of patients with GD. A recent observational study in Italy reported that the clinical phenotype of GD is becoming milder, likely due to increased awareness on thyroid disease and the availability of sensitive diagnostic tests, in conjunction with timely and effective treatments [3]. However, cases of extreme hyperthyroidism surface occasionally. Additionally, current treatments for GD, including antithyroid medications, radioactive iodine, and surgery (thyroidectomy), often result in adverse effects. Therefore, the importance of improving the understanding of the molecular and intermolecular interactions and the key signaling pathways affected by Graves' disease persists.
The pathogenesis of Graves' disease remains insufficiently elucidated. However, studies revealed GD as a B cell-mediated, T cell-dependent autoimmune disease with multiple types of immune cell involvement [4]. In patients with GD, overproduction of thyroid hormones and thyroid hyperplasia are often caused by stimulatory autoantibodies against the thyroid-stimulating hormone receptor (TSHR), also known as the thyroid-stimulating antibody (TSAb) [5]. Over the course of GD, communication between different immune cells is of great importance [6]. Aside from direct cell-to-cell contact, a novel and perhaps even more complex mode of immune regulation is provided by exosomes that are released by dendritic cells, B cells, T cells, and other cell types [7]. With the diameters ranging from 30 to 150 nm, exosomes are small membrane vesicles of endocytic origin, which are secreted by almost all cell types, displaying diverse functions dependent upon their origin [8]. Exosomes are rich in content and contain protein, lipid, and nucleic acid, which are involved in immune responses, antigen presentation, intercellular communication, protein and RNA transport and other physiological processes [9]. Current research into the genetic etiology of GD suggests that the "hidden" heredity of GD is likely associated with rare genomic variants including noncoding RNAs (ncRNAs), microRNAs (miRNAs), and/or epigenetic factors [10]. The exosomal circular RNAs (circRNAs), rather than those unprotected ncRNAs in blood, potentially represent novel bridges for communication of various immune cells in GD.
The first circular RNA (circRNA) was identified as 'scrambled exons' more than 40 years ago [11], but it was only recently more and more evidence of their expression in various diseases have been emerging. Unlike traditional linear RNA, which contains 5' and 3' ends, circRNAs have unique structures resulting from a 3′ to 5′ end-joining event (backsplicing). Meanwhile, two different models of circRNA biogenesis have been described, the lariat or exon skipping model and the direct backsplicing model Meanwhile, two different models of circRNA biogenesis have been described, the lariat or exon skipping model and the direct backsplicing model. [12]. CircRNAs can be derived from exon, intron, untranslated or intergenic regions of the genome, and a large amount of them have been identified in mouse and human by high-throughput sequencing and bioinformatics analyses [13]. CircRNA is produced by special selective shear and is abundant in the cytoplasm of eukaryotic cells. It expresses with tissue, timing and disease specificity [14]. Though to the multiple functions of circRNAs recently inferred, the most important is the involvement of circRNAs in post-transcriptional regulation, similar to the roles of an endogenous RNA or miRNA sponge in competitively inhibiting RNA/miRNA transcriptional regulation [15, 16]. In addition, circRNA can also regulate variable shear or transcription and modify the expression of its related genes [17]. Therefore, circRNAs are expected to provide a new molecular basis of prediction, diagnosis and treatment of complex diseases. However, the means and extent to which exosomal circRNAs play a role in the pathogenesis of GD remains completely unknown.
In this study, the implications of circRNA in GD and exosomal circular RNAs were profiled in the serum samples from patients with and without GD through human circRNA microarray. Comprehensive bioinformatics analyses were conducted to explore and probe the functions of the differentially expressed circRNAs.
Validation of study cohorts and isolated exosomes
This study involved two cohorts of subjects: the discovery cohort and the validation cohort. The clinical characteristics of the participants in the discovery cohort are summarized in Additional file 1: Table S2. The mean age of patients with Graves' disease was 36.6 ± 11.5 years as compared to mean age of 37.8 ± 9.6 years among controls (P = 0.86). As expected, Graves' disease patients had significantly elevated FT3, elevated FT4, and suppressed TSH levels in comparison to controls (Additional file 1: Table S2). Notably, the concentration of plasma TRAb (TSH receptor antibodies, also called TSAb), the specific thyroid autoantibody produced in GD (29.49 ± 14.77 IU/L), was remarkably higher in GD patients than in healthy control subjects (0.55 ± 0.32 IU/L). In the validation cohort (Additional file 1: Table S3), there is no difference in the age of patients with Graves' disease (37.55 ± 12.18 years) and control groups (37.15 ± 10.89 years) (P = 0.91). TSH(GD group: 0.0145 ± 0.0294 IU/ml vs. control group: 1.851 ± 0.8749 IU/ml), FT4(GD group: 90.66 ± 10.71 pmol/L vs. control group: 16.32 ± 2.06 pmol/L), FT3(GD group: 38.2 ± 10.83 pmol/L vs. control group: 4.85 ± 0.93 pmol/L), had a significant differences between two groups(P < 0.0001). There are significant differences in TRAb between GD (26.13 ± 12.9 IU/L) and healthy group (0.71 ± 0.46 IU/L) (P < 0.0001). The median TgAb level of GD and healthy control groups were 1887(127.6–4000) IU/ml and 20.82(10–114.1) IU/ml, respectively (P < 0.0001). Next, we verified isolated exosomes through observation of their morphology by transmission electron microscopy. As shown in Additional file 1: Figure S1, these exosomes displayed irregular spheres with a clearly defined and relatively intact membrane. They have diameters ranging from 30 nm to 100 nm, distinguishable from other much larger extracellular vesicles like microvesicles and apoptotic bodies.
Identification of differentially expressed plasma exosomal circRNAs from patients with GD and healthy control subjects
The plasma exosomes isolated from the discovery cohort were subjected to the microarray profiling of the differentially expressed circRNAs. A number of the 13,000 total circRNAs detected, developed an altered expression. As shown in the scatter plot, denoting the means of the expression values of the two groups (Fig. 1a), many circRNAs, from patients with GD, exhibit more than a twofold up-regulation or down-regulation of plasma exosomes compared to healthy control subjects. There are 15 significantly differentially expressed circRNAs that met the criteria of exhibiting fold changes > 1.2 and P-values < 0.05, as shown in the volcano plot representation of differentially expressed transcripts and their statistical significance (Fig. 1b). Among them, six circRNAs were up-regulated and nine circRNAs were down-regulated (Table 1).
Differential expression of circRNAs in plasma exosomes from patients with GD and healthy control subjects. a Scatter plots with the mean expression values of samples in each group. Raw junction reads for all samples were normalized by total mapped read numbers and were transformed by log2. Each dot represents a single circRNA. The exhibiting fold change is 2.0 of Line 1 and Line 3, whereas the dots on Line 2 indicate circRNAs with equal expression between GD group and healthy control group. b Volcano plot representation of differentially expressed transcripts and their statistical significance. The fold changes are represented in log2 scale as depicted on the x-axis, whereas the − log10 P-value is depicted on the y-axis. Transcripts with greater statistical significance are higher in the plot. The red circles represent circRNA genes that show differential expression with fold changes > 1.2 and P-values < 0.05 between the GD and normal control exosomes samples
Table 1 The list of differential expressed circRNAs in plasma exosome samples from patients with GD and healthy control subjects
hsa_circRNA_000102 is the validated differentially expressed plasma exosomal circRNA in GD
Among 15 significantly differentially expressed circRNAs, 12 are exonic, two are intronic, and one is a combination of both (Table 1). We examined the genes associated with the back splicing-mediated generation of these circRNAs, and found MED1, AKNAD1, PPWD1 and IL4R were related to nuclear co-receptor, transcription factor, mRNA splicing and cytokine receptor pathways, respectively. These molecules and pathways possess the ability for an immune activation, as observed in patients with GD. Therefore, the corresponding four circRNAs, including two up-regulated circRNAs (hsa_circRNA_000102, hsa_circRNA_102059) and two down-regulated circRNAs (hsa_circRNA_004939, hsa_circRNA_072697), were initially selected for further verification by RT-qPCR. As shown in Fig. 2 , the consistency between RT-qPCR results and microarray analysis data had been demonstrated only for hsa_circRNA_000102. However, the expression levels of hsa_circRNA_102059, hsa_circRNA_004939, and hsa_circRNA_072697 did not show significant differences between plasma exosomal samples from patients with GD and healthy control subjects (Fig. 2a–c). Furthermore, it was observed that expression levels of hsa_circRNA_000102/β-actin(r = 0.5983, P = 0.0053), hsa_circRNA_000102/18srRNA(r = 0.5462, P = 0.0127), hsa_circRNA_000102/ GAPDH(r = 0.4526, P = 0.0451) correlate significantly with TRAb levels (Fig. 2d–f). Therefore, our next stage of bioinformatics analyses focused on delineating the intermolecular interactions and the key signaling pathways associated with hsa_circRNA_000102.
Validation of selected candidate differentially expressed circRNAs by Real-time qPCR. Data represents the mean ± SD of three independent experiments. n = 10 for each group. *P < 0.05
Prediction of the circRNA/microRNA/mRNA interaction network for hsa_circRNA_000102
Recent studies on circRNA/microRNA interactions have indicated that circRNAs play a key role in the regulation of gene expression by interacting with their target microRNAs (miRNAs) [18]. To find the potential target miRNAs and mRNA associated with GD, a circRNA/microRNA/mRNA interaction network was constructed to predict the intermolecular interaction for the experimentally confirmed circRNA (hsa_circRNA_000102). An Arraystars homemade miRNA target prediction software based on TargetScan and miRanda was utilized (Fig. 3). There were nine miRNAs (hsa-miR-3151-5p, hsa-miR-1227-5p, hsa-miR-194-3p, hsa-miR-1296-3p, hsa-miR-3688-3p, hsa-miR-7112-3p, hsa-miR-8063, hsa-miR-4512 and hsa-miR-7848-3p) and numerous mRNAs predicted to be interacting with hsa_circRNA_000102 directly or indirectly.
The circRNA-miRNA-mRNA regulatory network of hsa_circRNA_000102. CircRNA, miRNA, and mRNA are indicated as spheres in brown, red, blue color, respectively. CircRNA circular RNA, miRNA microRNA, mRNA messenger RNA
GO analysis and KEGG pathway analysis of hsa_circRNA_000102 associated genes
Predicted genes in the circRNA/microRNA/mRNA interaction network of hsa_circRNA_000102 were used for GO function analysis to annotate and speculate the potential functions of this circRNA. GO analysis was divided into three parts: biological process (BP), cell component (CC) and molecular function (MF) (Fig. 4). GO analysis of BP showed that hsa_circRNA_000102 was significantly linked to Poly(A)+ mRNA export from nucleus, cellular response to interferon-beta, mRNA-containing ribonucleoprotein complex export from nucleus, mRNA export from nucleus, RNA transport, nucleic acid transport, regulation of production of molecular mediator of immune response, establishment of RNA localization, and positive regulation of cytokine biosynthetic process. Significant GO cell component terms of hsa_circRNA_000102 showed that it was associated with endocytic vesicle lumen, endolysosome membrane, Cul2-RING ubiquitin ligase complex, endolysosome, high-density lipoprotein particle, lipoprotein particle, plasma lipoprotein particle, cytoplasmic vesicle lumen, vesicle lumen, and ubiquitin ligase complex. For molecular function, hsa_circRNA_000102 was associated with transition metal ion binding, receptor ligand activity, phosphatidic acid binding, receptor regulator activity, protein transporter activity, metallocarboxypeptidase activity, tumor necrosis factor receptor binding, chemoattractant activity, molecular function regulator, and Zinc ion binding. The top 10 KEGG pathways showed that hsa_circRNA_000102 associated genes might be involved in Herpes simplex infection, Influenza A, RNA transport, mRNA surveillance pathway, Ribosome biogenesis in eukaryotes, Hepatitis B, steroid hormone biosynthesis, necroptosis, NOD-like receptor signaling pathway and Huntington disease (Fig. 5).
The top 10 significantly enriched terms of biological process by GO analysis of hsa_circRNA_000102 associated genes. GO analysis was divided into three parts: biological process, cell component and molecular function
The top 10 significant enriched pathways by KEGG pathway analysis of hsa_circRNA_000102 associated genes
In this study, we profiled the plasma exosomal circRNAs from patients with primary GD and healthy control subjects by human circRNA microarray. Before circRNA labeling and hybridization, RNAse R is used to efficiently digesting nearly all linear RNA species. Although RNAse R treatment may not exclude all entities, it is one of few ways at current available for enriching circRNAs. Our initial screening resulted in 15 differentially expressed circRNAs. One of the four selected candidate circRNAs, hsa_circRNA_000102, was verified to be significantly enriched in the plasma exosomes of patients with GD. Our comprehensive bioinformatics analyses, including prediction of a circRNA/microRNA/mRNA interaction network for hsa_circRNA_000102 and GO and KEGG pathway annotations of hsa_circRNA_000102 associated genes, revealed potential pathways correlated to the pathogenesis of GD.
hsa_circRNA_000102 is a circular intronic RNA (ciRNA), a new type of circRNA in human cells that is derived from introns discovered by Zhang et al. and others [19]. These RNAs escape debranching and depend on consensus RNA elements near the 5' splice site and the branchpoint for proper processing [19]. Interestingly, ciRNAs partially accumulate to their sites of synthesis and colocalize with the elongation RNA polymerase II (Pol II). Moreover, the existence of ciRNAs might influence the transcription of their parent genes. For example, they are specifically associated with phosphorylated Pol II whereby a depletion of ciRNAs may lead to a significant reduction in the transcription of parent genes. Therefore, the higher level of hsa_circRNA_000102 in plasma exosomes of patients with GD, indicates the potentially elevated expression of its parent gene, AKNAD1, which encodes a protein with a domain found in an AT-hook-containing transcription factor. Though being not yet well-characterized, AKNAD1 is considered to involve in biological processes of "microtubule-based movement" and "microtubule-based process". Till now, the relation between gene AKNAD1 and circRNA has not been discussed. Only one research has identified a significant copy number variation in gene AKNAD1 which was validated to be associated with type-2 diabetes through stress of endoplasmic reticulum [20], suggesting a potential link of AKNAD1 and autoimmune diseases. However, more work on AKNAD1 in immune regulation and the underlying mechanisms on potential immune activation in GD is required to delineate the relationship between AKNAD1 expression and the pathogenesis of GD.
The roles of circRNAs have been identified in tumor signaling pathways, invasion and migration, as well as tumor immunity [20]. Previous research proposed that mRNAs, transcribed pseudogenes, and circRNAs communicate with and co-regulate each other through competition for binding to the shared miRNAs [21]. Studies have also revealed that the activity of circRNA is decided by a series of factors, including the abundance of miRNAs, RNA editing, changes in the RNA 3′ untranslated region (UTR) and RNA binding proteins (RBPs) [22]. Therefore, interpreting the cross talks among circRNAs, miRNAs and mRNAs would provide novel insights into the mechanisms of gene regulation and immune cell interaction in Graves' disease. We predicted the circRNAs/miRNAs/mRNAs interaction network of hsa_circRNA_000102, and then performed GO enrichment analysis and KEGG pathway analysis of the particular circRNA associated genes. Evidence demonstrated that hsa_circRNA_000102 is related to Herpes simplex infection, Influenza A signaling and interferon-beta signaling.
Both genetic and environmental factors have been suggested to contribute to the epidemiology of Graves' disease [23]. It is not known whether the infection of a Herpes virus or Influenza A and Graves' disease could co-exist simply by chance or the former could trigger the latter. A previous study has detected Herpes virus DNA in post-operative thyroid tissue specimens of patients with autoimmune thyroid disease, including Graves' disease and Hashimoto thyroiditis [24]. The possible role of human Herpes viruses in the pathogenesis of autoimmune thyroid diseases is largely unknown and data from one study did not support their association [25]. However, another study indicated that Graves' disease is linked to infectious mononucleosis due to primary Epstein-Barr virus infection in patients [26]. This suggests that both Herpes virus and Influenza A virus can employ similar mechanisms as Epstein–Barr virus to trigger immune activation.
We speculate that the association between viral infection and GD is due to the efficient T cell activation. Viral infection potentially increases the efficiency of T cell activation through fast attraction of dendritic cells from peripheral to secondary lymphoid tissues. Furthermore, continuous activation of innate immunity against viral infection might also profoundly strengthen autoimmune disorders by promoting autoantigen release without the need for specific activation of autoreactive T cells. As previously reported [27, 28], thyroid autoimmunity and dysfunctions have been observed over the course of IFN-beta therapy. A patient with multiple sclerosis received IFN-beta-1 therapy for 22 months and ultimately developed a severe Graves' disease with persistently positive TRAb [27]. Since thyroidal side effects can be late to diagnose, patients undergoing long-term IFN-beta therapy should be closely monitored for thyroid hormones and antibodies throughout the treatment to prevent the possible occurrence of GD [27]. Taken together, although further studies are needed to clarify the relationship between infection of Herpes virus/Influenza A or IFN-beta signaling and GD, our study indicates that the impact of virus infection and IFN signaling should be taken into consideration during the prevention and/or treatment of GD.
The current investigation has several strengths and limitations. Its major strength is its novelty; to the best of our knowledge, the current study is the first one on profiling of plasma exosomal circRNAs in patients with GD. Our work also provides a feasible framework for searching and validating circular RNAs to better facilitate target identification and development of target-oriented therapeutics. The major limitation of the current study is its sample size of patients used for discovery and validation. Moreover, the patients recruited in this study were from a single institution and thus are subject to referral bias. Thirdly, there are some other limitations in this study. For examples, ribosomal RNAs were not removed, and the expression of AKNAD1 was not measured. In addition, no mock-treated samples were used during samples prep stage for generation of circRNA profile. Also, circRNA arrays we used is based on backsplice junctions pre-designed for those already existing in database. Therefore, this profile is very likely to lack novel circRNAs that could have been discovered using alternative methods to generate circRNA profiles.
In this study, we performed human circRNA microarray in the plasma exosomes from patients with GD and healthy control subjects and verified hsa_circRNA_000102 as a differentially up-regulated plasma exosomal circRNA in GD. Our comprehensive bioinformatics analyses including construction of circRNA/microRNA/mRNA interaction network, GO annotation and KEGG pathway analysis, highlighted multiple pathways, particularly the virus infection and interferon-beta signaling, in mediating immune activation in Graves' disease.
Between August 2017 and October 2017, five newly diagnosed patients with GD (two males and three females) and five healthy control subjects were recruited by the Affiliated Yantai Yu-huangding Hospital of Qingdao University Medical, Shandong Province, China. The newly diagnosed GD patients had typical characteristics of hyperthyroidism: a palpable, soft, and diffuse goitre confirmed by ultrasound examination, increased thyroid radioiodine uptake, elevated serum free thyroxine (FT4), very low thyroid-stimulating hormone (TSH), and high levels of thyroid-stimulating antibody (TSAb). Age- and gender-matched healthy control subjects with normal thyroid function (TSH: 0.27–4.2 mIU/L), free of goitre and any family history of GD who meeting the exclusion criteria selected from healthy hospital staff were admitted into the study. Exclusion criteria included patients with liver disorder, renal disorder, clinical evidence of tumor, infection, any autoimmune diseases, history of intake of immunosuppressive drugs, such as mannan peptide, lentinan, interferon, corticosteroids, and cyclophosphamide, within 1 year of the study, pregnant, and lactating women. In the validation cohort, 20 patients with GD and 20 healthy control subjects were recruited adhering to the same inclusion and exclusion criteria. The research protocol was approved by Medical Ethics Committee of Affiliated Yantai Yu-huangding Hospital of Qingdao University Medical and the written informed consent was obtained from all participants.
The levels of serum TSH (11731459, Roche), FT4 (07976836, Roche), FT3 (free triiodothyronine) (06437206, Roche), TSAb(04388780190, Roche), thyroid peroxidase antibody (TPOAb) (06368590190, Roche), and thyroglobulin antibody (TgAb) (06368697, Roche) were measured via chemiluminescent enzyme immunoassay specific kits (Cobas 6000, Roche).
Ultracentrifugation exosomes isolation and RNA extraction
Venous blood samples from subjects were collected in evacuated tubes with the anticoagulant Ethylenediamine tetraacetic acid dipotassium (K2EDTA) and stored at − 80 °C for exosome isolation. All blood samples were processed by centrifugation at 1000×g for 10 min. The plasma was immediately separated and transferred to a new RNase free tube. The liquid samples were centrifuged at 500×g for 5 min to eliminate cells. The supernatant was then transferred to a new polycarbonate tube and centrifuged at 2000×g for 10 min. Supernatants from the previously centrifuged samples were collected and centrifuged again at 10,000×g for 30 min to eliminate shed microvesicles (sMV, 200–1000 nm). The supernatants were then collected, filtered with 0.22 μm membrane filter (Merck Millipore), and centrifuged at 100,000×g for 2 h to obtain exosomes. For RNA isolation, an exosome pellet was washed once with 1×PBS and centrifuged at 100,000×g for 2 h. Exosomes were observed under a transmission electron microscope (TEM, TecnaiG2 Spirit 120 kV). Total RNA was extracted from exosomes using the TRIzol LS Reagent (10296028, Invitrogen Life Technologies, Carlsbad, CA, USA) according to the manufacturer's protocol.
Linear RNA digestion, circRNA labeling and hybridization
The exosomal RNAs extracted from the discovery cohort (five GD patients and five healthy control subjects) were used for microarray analysis, as previously described [29]. RNase R (RNR07250, Epicentre, Inc.) is used to digest nearly all linear RNA species and enrich circRNAs. The circRNA expression microarray slide (AS-S-CR-H-V2.0, Arraystar Human circRNA Arrays V2, 8 × 15 K; Arraystar Inc.) was used. Sample labeling and array hybridization were performed according to the manufacturer's protocol. The hybridized arrays were scanned using the Agilent Scanner G2505C (Agilent Technologies, USA).
CircRNA microarray data analysis
Raw data extraction of the scanned images was conducted using the Agilent Feature Extraction software (version 11.0.1.1). A series of data processing, including quantile normalization using a log2 ratio, was performed using the R software limma package. The circRNAs containing flags in "P" or "M" in at least five out of 10 samples (defined by GeneSpring software) were retained for further differential analyses. Differentially expressed circRNAs displaying statistically significant differences between the GD and healthy control groups were identified using fold change cutoff and through Volcano Plot filtering. The "fold change" (i.e. the ratio of the group averages) between the groups for each circRNA was computed. A t-test was employed to determine the statistical significance of the differences measured. CircRNAs denoted as significantly differentially expressed, were those having fold changes > 1.2 and P-values < 0.05 between the GD and healthy control exosome samples. The package gplots and function heatmap2 in R software were used for mapping. The distance metric was set as Euclidean distance.
Real-time quantitative PCR (RT-qPCR) validation of microarray data
Total RNA from each exosome sample was quantified using the NanoDrop ND-1000 spectrophotometer. The reverse-transcription (RT) primers for circRNAs and β-actin were designed using Primer 5.0 and listed in Additional file 1: Table S1. Complementary DNA (cDNA) was synthesized using SuperScript™ III Reverse Transcriptase (18080-044, Invitrogen). CircRNAs transcripts were quantified using a SYBR Green Real-time PCR Master Mix (AS-MR-006-25, Arraystar, US). PCR was performed with the ViiA 7 Real-Time PCR System (Applied Biosystems). The thermal cycler conditions consisted of one cycle of denaturation at 95 °C for 10 min followed by 40 cycles of 95 °C for 10 s and 60 °C for 60 s. Melting curves were generated to ensure the specificity of the RT-qPCR. The expression of circRNAs was calculated by standard curve method using β-actin, 18srRNA and GAPDH as housekeeping genes. Then we compare the relationship among circRNAs and TRAb. All reactions were performed in triplicate.
Annotation for circRNA/miRNA Interaction
RNA transcripts crosstalk by competing for common microRNAs and have been termed as competing endogenous RNAs (ceRNAs) [30]. Studies identified microRNA response elements (MREs) as the foundation of this interaction [31]. CeRNAs include pseudogene transcripts, long non-coding RNAs (lncRNAs), circRNAs and mRNAs, all of whose transcripts can compete for the same MRE to regulate mutually. Potential microRNA targets were identified with home-made miRNA target prediction software based on TargetScan and miRanda [32, 33]. The ceRNA network was constructed through merging the common targeted miRNAs. Three conditions must exist for the ceRNA network to occur, as previously described, which include: well defined relative concentrations of ceRNAs and their microRNAs; an appropriate number of microRNAs that ceRNA can "sponge"; and knowledge that not all of the MREs on ceRNAs are equal [30]. Therefore, only the ceRNA-pair relations passing these measures were accepted as filtered. The co-expression network was created according to the correlation analysis between normalized intensity of the differential circRNAs, miRNAs and mRNAs. Pearson correlation coefficients over 0.95 were retained for the construction using CYTOSCAPE software (The Cytoscape Consortium, San Diego, CA).
Bioinformatics analysis
Gene ontology (GO) analysis was performed to explore the functional roles of circRNA-targeting genes in terms of biological processes, cellular components and molecular functions, as previously described [34]. Biological systems defined by the Kyoto Encyclopedia of Genes and Genomes (KEGG) (http://www.genome.jp/kegg/), was used to explore those pathways related to circRNA-targeting genes, as previously described [35].
SPSS software, version 17.0 (SPSS Inc.) was used for statistical analyses. Data are expressed as the mean ± SD (standard deviation) or median values with ranges, as indicated. An unpaired two-tailed Student's t-test, or a nonparametric Mann–Whitney test was used for the comparisons between groups. Welch's correction was added when the variances were significantly different between the two groups. Pearson's correlation coefficient was used to analyze correlation between hsa_circRNA_000102 and TRAb. A two-sided P value < 0.05 was considered to be statistically significant.
All data generated or analysed during this study are included in this published article.
circRNA:
Circulating RNA
GD:
Graves' disease
DEcRs:
Differentially expressed circRNAs
KEGG:
Thyroid-stimulating hormone receptor
TSAb:
Thyroid-stimulating antibody
ncRNAs:
Noncoding RNAs
miRNAs:
TRAb:
TSH receptor antibodies
ciRNA:
Circular intronic RNA
RBPs:
RNA binding proteins
TPOAb:
Thyroid peroxidase antibody
TgAb:
ceRNAs:
Endogenous RNAs
MREs:
microRNA response elements
lncRNAs:
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Thomas D, Liakos V, Michou V, Kapranos N, Kaltsas G, Tsilivakos V, et al. Detection of herpes virus DNA in post-operative thyroid tissue specimens of patients with autoimmune thyroid disease. Exp Clin Endocrinol Diabetes. 2008;116:35–9.
Noura AZ, Monem F. Are human herpes viruses associated with autoimmune thyroid disease? J Infect Dev Ctries. 2011;5:890–2.
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Schwid SR, Goodman AD, Mattson DH. Autoimmune hyperthyroidism in patients with multiple sclerosis treated with interferon beta-1b. Arch Neurol. 1997;54:1169–90.
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I would like to show my deepest gratitude to Nana Huang and Fangfang Wei for all their work. I thank all my friends for their encouragement and support.
This project was supported by the National Natural Science Foundation of China (No. 81700695) and China Scholarship Council (No. 201909370036).
Ying Sun, Wei Wang, Yuxiao Tang, Daping Wang and Jin Zhou contributed equally to this paper
Department of Endocrinology, Affiliated Yantai Yuhuangding Hospital of Qingdao University Medical College, Yantai, Shandong, China
Ying Sun, Wei Wang, Yuxiao Tang, Daping Wang, Shulin Chen & Jin Zhou
Department of Neurosurgery, Affiliated Zhongshan Hospital of Dalian University, Dalian, Liaoning, China
Liang Li
Department of Radiology, Dalian Sixth People's Hospital, Dalian, Liaoning, China
Min Na
Department of Endocrinology and Metabolism, Binzhou Medical University Hospital, Binzhou, Shandong, China
Guantong Jiang
Department of Scientific Research, Affiliated Yantai Yuhuangding Hospital of Qingdao University Medical College, Yantai, Shandong, China
Qian Li
Ying Sun
Yuxiao Tang
Daping Wang
Shulin Chen
Jin Zhou
JZ and SC designed the research. YS, WW, YT and DW conducted the experiments, analyzed data, and wrote the manuscript. QL contributed to the study design and manuscript writing. MN edited and revised manuscript. LL analyzed data. GJ performed experiments. All authors read and approved the final manuscript.
Corresponding authors
Correspondence to Qian Li or Jin Zhou.
The present study was approved by Medical Ethics Committee of Affiliated Yantai Yu-huangding Hospital of Qingdao University Medical and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Additional file 1.
Additional figure and tables.
Sun, Y., Wang, W., Tang, Y. et al. Microarray profiling and functional analysis of differentially expressed plasma exosomal circular RNAs in Graves' disease. Biol Res 53, 32 (2020). https://doi.org/10.1186/s40659-020-00299-y
DOI: https://doi.org/10.1186/s40659-020-00299-y
Circular RNA
hsa_circRNA_000102
Immune activation


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Greg Galitzine
Greg Galitzine's VoIP Authority Blog
| VoIP and IMS News and Views
You are browsing the archive for July 2005.
Top 10 VoIP Providers Named
I came across an entry from Russell Shaw today listing the Top 10 VoIP Providers by subscription count. The Top 10 is based on research provided by Point Topic.
Skype - 5,300,000
Yahoo! Japan - 4,517,000
VoiceGlo - 605,000
Free (France) - 600,000
Vonage - 535,000.
FastWeb (Italy) - 528,000
Cox - 413,000
Time Warner - 372,000
Cablevision - 364,000
Neuf (France) - 297,000
According to Point Topic, there were over 11 million people using retail VoIP service for at least some of their telephone calls at the end of March 2005.
Brightpoint Signs Reseller Agreement With Vonage
Vonage announced an agreement with Brightpoint today, whereby the latter's Brightpoint Activation Services LLC will become a Master Agent, offering its customers the opportunity to resell Vonage service plans and� devices and also to activate new customer accounts.
Vonage continues to expand its distribution channel. Pretty soon, one might be able to drive up to a gas station and receive offers for free Vonage calling with every fill-up.� I don't think next-generation telecom services can ever replace promotional glasses as a primo giveaway, but heck... I'm always the last one to hop on a trend.
Merrill Lynch Releases Telecommunications Report
Merrill Lynch released a report today entitled, Telecommunications: US Telecom 2Q05 Preview.
� Among the findings released today, Merrill Lynch offers a series of expectations for the second quarter. Quoting directly from the report:
The seasonal trends in telecom for the second quarter period favor wireless over wireline, even more so than for other periods, due in part to demographic drivers. We expect this to be the case in 2Q 05.
We think that Verizon Wireless will gain share during 2Q 05, in another solid quarter of wireless results.
And now, the financial report....
I can just hear the old-style ticker in the background (deet deeet deetde deet...) this just in...!
Crown Castle International Corp. has made an additional investment of $55 million in FiberTower Corporation, a privately-held provider of backhaul services.
FiberTower was a recent recipient of TMC Labs' 2005 Innovation Award. You can see the full write-up in the July issue of Internet Telephony magazine.
German Unit To Block VoIP Applications
Seems that VoIP is scaring the scheize out of some folks. According to a published report this morning, Vodafone's German unit has made plans to disable calls from VoIP operators such as Skype and others beginning in July 2007.
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The article quotes Vodafone Germany spokesman Heiko Witzke, who said that in the interim the company may yet reverse its policy. Witzke wouldn't comment about why the company was taking the step, but said that "2007 is a long ways to go; anything may happen until then."
Hmm… Established, legacy carriers, fearful of losing market share to VoIP… That can't happen here, right?
TMC, CompTel/Ascent to Host Joint e911 Webinar
The question is no longer "When" but "How."
How will service providers comply with the recent FCC order mandating that all VoIP service providers must provide 911 services within 120 days. The 120-day clock is now ticking and VoIP service providers are working diligently to meet the deadline.
CompTel/ALTS, in partnership with TMC, is pleased to present a Webinar on the FCC's E911 Order and its impact on VoIP service providers.
The event is set to take place on Monday, July 18th at 1pm Eastern Time.
Back in Yellow
By the way, for those of you who may have missed it, Lance Armstrong has regained the yellow jersey in the wake of his strong performance yesterday. Today, the suffering continues as the peloton heads back into the Alps for a ride that features 2 HC (hors category) climbs, and another slightly less imposing Cat 1 climb. HC designates the most difficult mountain climbs.
I don't expect that Mr. Armstong will be relinquishing the yellow race leader's jersey any time soon.
Keep Your Bling-Bling... I Got Vling Vling!
EarthLink is rolling out Vling, a sparkling new service that integrates chat with voice through a single easy-to-use interface. Built on SIP, Vling currently runs on Windows XP and is available for free to all high-speed Internet users who want to connect on their terms via voice, data, or chat. The service is currently in beta.
Using peer-to-peer, file-sharing technology, Internet consumers can download Vling and begin making calls to other Vling users and customers of select other SIP-based services.
Over the next several months, EarthLink plans to further its position in the voice market by testing other innovative voice/VoIP technologies. Watch for them to get in to the dual-mode space with a dual-band WiFi/cell phone offering soon.
snom To Showcase Phones At VoIP Reseller Event
snom technology AG will demonstrate its full line of open-standard, SIP-based VoIP telephones as a sponsor of VoIP Sizzles in Dallas — held July 20-22 at the Sheraton Airport Grand, in Dallas, TX.�
I recently blogged about the upcoming� event.
According to a company spokesperson,� snom technology will be demonstrating its complete line of SIP standard-based VoIP telephones, including the recently-launched entry-level snom 320 and the large-display executive snom 360. snom's Dr. Michael Knieling, SVP Marketing, will be available throughout the event to consult with attendees and speak with media.
"We're excited to be sponsors of this unique event," says Dr. Knieling, "along with other key players in the fast-growing open IP telephony community.
Big Day For Discovery
Today is the day that� we will separate the men from the boys� at the Tour de France. In today's 10th stage, the first real mountain test in the Alps, expect Lance Aremstrong and his Discovery teammates to come under attack.
First and foremost, the German T-Mobile team� is not only placed well to apply pressure, they're also hungry to place one of their riders at the top of the general classification. Jan Ullrich, Alexandre Vinokourov, and Andreas Kloeden of the T-Mobile squad, are all legitimate contenders for the overall title, and today might be the day that one (or all) of them strike out for the top spot.
Alcatel Earnings Up
Alcatel has posted higher-than-expected revenue growth for the second quarter thanks to higher than expected revenue growth resulting from strong momentum in IP routing and optical transmission, mobile communications, and enterprise solutions.
Announcing figures today, France-based communications gear maker reported Q2 revenues were up 8 per cent to €3.14Billion (~$3.82Billion).
Radicati Group Promotes Unified Communications Report
The Radicati Group unveiled their latest report today, entitled On-Premises Corporate Unified Communications Market, 2005-2009. The research report provides market size, four-year forecasts, technology trends, and competitive information for the On-Premises Corporate Unified Communications market. The report also includes installed base data broken out by geography and size of business.�
The Radicati report forecasts healthy growth in the market over the next four years, largely driven and supported by emerging technologies such as VoIP, SIP, and a range of access technologies.
Hot Off The Presses!! Internet Telephony Conference & EXPO Brochure!
The Internet Telephony Conference & EXPO show brochure is now available!
You can also download the conference grid at the site.
The program at this Fall's event was designed� with the entire VoIP industry in mind -- if you are in search of unbiased, educational information, then Los Angeles is the only place you need to be to learn about VoIP this October.
Conference registration is open to all - the registration page can be accessed by clicking here.
Goldman, Google, Hearst Invest IN Broadband Over Power Line (BPL) Provider
CURRENT Communications Group, LLC, a Broadband over Power Line (BPL) service provider, today announced strategic investments from Goldman, Sachs & Co., Google Inc., and The Hearst Corporation.
"These investments provide us with both capital and operating assistance as we continue to roll out Broadband-over-Power Line services to provide voice, video and data services," said William Berkman, Chairman of CURRENT Communications Group. "With roughly two-thirds of the nation still without broadband and many utilities looking for means to upgrade their distribution networks, the acceleration of CURRENT's build-out is a clear win-win for consumers."
BPL technology enables the delivery of high-speed, two-way broadband through electrical outlets in homes and offices.
RNK Introduces EDISON: GPS VoIP, E911 Solution
RNK Telecom has announced the launch of Edison — a GPS-enabled solution that is designed to connect between a user's phone and VoIP connection, ensuring E911 systems continuously receive precise and updated information about the VoIP caller's location.
RNK co-founder and CEO Richard Koch said, "Edison is one of many products RNK is pouring R&D funds into. We've been testing Edison for many months now, and it will prove that RNK isn't just a phone company. Edison can save the lives of VoIP callers."
RNK's announcement comes just five weeks after the FCC mandated that all VoIP providers must provide enhanced 911 services within 120 days.
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CINCINNATI, OH - SigmaTEK named four additions to its Central United States Region team. The team works under regional sales manager, Mike Galetti, regional sales manager and a 23-year veteran of the fabrication industry.
"As manufacturing across the board continues to ramp up companies are taking a long look at tools, technologies, and processes in an attempt to maximize production and eliminate waste. Nowhere is this more evident than in the industrious central states," says Galetti. "We are taking these aggressive steps to remain responsive to the needs of both current and future customers."
Todd Shrum, regional technical manager: with 16 years manufacturing experience, Shrum teamed up with SigmaTEK in the development of combination punch plasma machines while working at WA Whitney as an applications manager. He assists in SigmaNEST implementation to fully leverage software capabilities for multiple cutting processes and related manufacturing applications.
Jennifer Gallagher, regional sales representative: a U.S. Army veteran, Gallagher stepped into manufacturing as a SigmaNEST programmer for Tanaka, Mazak, and Mitsubishi laser machines. She now works with implementing 3D CAD into SigmaNEST, based in the Chicago, IL area assisting companies in utilizing material costs and improving shop floor efficiency.
Michel Scavuzzo, regional sales representative: Scavuzzo works with companies to evaluate advanced nesting and material utilization. Based in Dallas, TX, Michel and Chris Wickliffe work as a team to increase response times for business transactions and improve regional support.
Chris Wickliffe, regional applications engineer: a long-time SigmaNEST, Wickliffe holds a degree in mechanical engineering, and helps companies implement time, material and cost-saving programs. His background includes programming for punch, combos, laser, oxyfuel, plasma, waterjet, knife, routers and brake presses. Chris works out of the Dallas, TX office.


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Coming Soon to the Office: Facebook at Work
A screenshot from Facebook's new service aimed at professionals called Facebook at Work. (Courtesy: Facebook)
Coming Soon to the Office: 'Facebook at Work'
Facebook recently estimated that each of its users spends an average of 50 minutes a day on the company's online social network.
But the company wants to find ways to increase the time spent on Facebook and two other services, Instagram and Messenger.
A new product aimed at capturing our interest is called Facebook at Work.
Some businesses bar workers from using Facebook (openly) on the job. Now those individuals actually can use the service -- at least for work purposes.
Facebook at Work is separate from a users' personal Facebook. So people will need to sign up with the company for a new account. The idea, however, is exactly the same as Facebook – to connect as many users as possible, this time in the workplace.
Same Facebook look
The look and feel of Facebook at Work is just like the old Facebook. There are groups to join, a News Feed to read, and instant messaging to stay in contact with co-workers.
Naturally, users can add and share comments, photographs, videos and information about the company and its offices. "A connected workplace is a more productive workplace," explains the official Facebook at Work page.
Facebook at Work was first announced about two years ago. Since then, the company has been testing the service with several large companies.
A screenshot from Facebook's new app aimed at professionals called Facebook at Work. (Courtesy: Facebook)
One of those businesses is the Royal Bank of Scotland. Its Director of Design, Kevin Hanley, told Business Insider the service has been helpful to the bank's 100,000 employees.
"I think Facebook lets us communicate, discuss and solve problems that other solutions, such as email, simply can't."
Hanley told the website that within his company, 90 percent of employees who tried Facebook at Work remained active from month to month.
Faces much competition
Facebook at Work will be competing with other workplace products or services already in use. These include LinkedIn, Slack, HipChat, Microsoft's Yammer and Salesforce's Chatter.
According to the technology website TechCrunch, Facebook at Work will be launching in London on October 10. The site said it had seen invitations to the launch event that have been sent out.
Facebook has not yet confirmed a date for launching the service.
I'm Bryan Lynn.
Bryan Lynn wrote this story for VOA Learning English, based on information from Facebook, and reports from TechCrunch and Business Insider. George Grow was the editor.
We want to hear from you. Would you like to use Facebook at Work in your workplace? Write to us in the Comments section, and visit our Facebook page.
network – n. group of people who know each other or work together
account – n. agreement allowing a person to use a particular website or service
New Tech Tools Might Help Americans Choose a President
Video Calling Services Compared
Facebook CEO Gives Pope Drone During Vatican Visit


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You are not logged in. [Log In] JakChat » Forums » News, Opinion & Current Events » Business & Investment News » AT/Jakarta eyes palm oil for fuel
#32096 - 17 May 06 20:33 AT/Jakarta eyes palm oil for fuel
From Asia Times http://www.atimes.com/atimes/Southeast_Asia/HE18Ae02.html
Jakarta eyes palm oil for fuel
By Bill Guerin
JAKARTA - Spiking global oil prices have fuel-importing countries everywhere examining the potential of lower-cost biofuels to save on their fuel bills and shore up national energy security. That's particularly true for Indonesia, which currently imports 30% of its energy needs but also boasts Asia's biggest bounty of the tropical resources used to produce the most efficient biofuels.
Government officials are preparing plans to make Indonesia a major player in the still-nascent global biofuel industry, which if oil prices stay high will have enormous export potential. For years, the government has backed research into biofuels, including various biotechnological research and development projects through the state-run Agency for Assessment and Application of Technology (BPPT).
But only after the government last year slashed fuel subsidies did the private sector aggressively take up the biofuel cause. Speculation in Indonesia's biofuel potential has recently driven the share price of the country's main publicly listed producer of crude palm oil (CPO), Astra Agro Lestari, to all-time highs.
Plantation company PT Bakrie Sumatera Plantations, meanwhile, plans to build the country's first privately owned bio-diesel plant in a US$25 million joint venture with construction firm PT Rekayasa Industri. The plant will have a capacity of 60,000-100,000 tons of bio-diesel a year and will adopt a "multi-feed" concept using several different raw materials, such as CPO and castor oil.
In hopes of sparking grassroots activity, the government has earmarked $33 million to build four pilot bio-diesel plants, meant to serve as national models for alternative-energy production. The plants in Kalimantan and Sumatra, with a combined annual capacity of 6,000 tons, will reportedly be operative by the end of this year.
A new government roadmap lays down hard targets for using less petroleum and more alternative energy sources in the future, mostly biofuels. The biofuel-promoting plan aims aggressively to reduce the country's use of petroleum, which currently represents 60% of national fuel consumption, down to 30% by 2025.
Indonesia is the world's second-largest producer of palm oil, the material most commonly used in producing bio-diesel, a biofuel that may be blended directly with conventional petroleum-based diesel. From an efficiency perspective, the yield from Indonesia's CPO plantations is way ahead of other tropical biofuel options, including the use of coconut oil.
Because bio-diesel contains veritably no sulfur, it proponents note that bio-combustion produces few of the harmful oxides-of-sulfur emissions that petroleum-based fuels do. Moreover, the modification of most diesel-burning engines to use biofuels is a straightforward process.
Global demand for biofuels is surging, particularly in fuel-strapped, environmentally conscious Western Europe. European Union member countries are increasingly chasing up global supplies of bio-diesel to meet stringent European emission standards. European bio-diesel production meets a mere one-sixth of current demand, which is projected to grow much higher as long as global petroleum prices hover at or above US$60 per barrel.
Barriers to burn
While Indonesia's biofuel future may initially look bright, there are still plenty of barriers to burn before the country maximizes its potential. High initial investment costs, an absence of state incentives for growers and producers, and the lack of credit facilities for small-scale entrepreneurs all threaten to stymie the industry's development.
Moreover, the government has a history of squandering its natural resource endowments. Indonesia, long plagued by mismanagement and chronic official corruption, is the only member of the Organization of Petroleum Exporting Countries (OPEC) that despite its huge resources is actually a net importer of fuels.
President Susilo Bambang Yudhoyono is moving to lower at least some of the hurdles. A recent Presidential Instruction for the provision and utilization of biofuel sets out new parameters for its supply and usage - although details governing the policy's actual implementation are thin. The instruction will be incorporated into a draft law on national energy due soon for debate in the House of Representatives.
Energy and Mineral Resources Minister Purnomo Yusgiantoro says the government is considering fiscal and administrative investment incentives for biofuel producers - although his plans simultaneously conjure up the specter of new bureaucratic tangles for the infant industry. Yusgiantoro said at a seminar this month that the Finance Ministry plans to formulate tax and import duty incentives for biofuel-production equipment.
Other ministries and governmental offices have been charged with formulating incentives other than plain fiscal ones, he said. For example, local administrations have been ordered to simplify arrangements for land-use permits, the Agriculture Ministry to encourage more raw-material production, and the Industry Ministry to simplify plant-licensing procedures.
If all the proposed incentives are actually adopted, the targets for biofuel production would steadily increase, starting at 720,000 kiloliters a year through 2010, ramped up to 1.5 million kiloliters per year from 2010 to 2015, and more than trebled at 4.7 million kiloliters per year in the decade ending in 2025, according to Yusgiantoro's projections.
Domestic bio-diesel retail sales in Indonesia are now estimated by his ministry at a mere one tonne per day. Indonesian regulators now allow fuel retailers to blend in up to 10% biofuel in their mixes. State-owned oil giant Pertamina recently unveiled two types of bio-diesel - known simply as B10 and B5 - in several fuel stations in Jakarta. Both fuels are supplied from the government's Agency for Assessment and Application of Technology, which currently runs a bio-diesel plant with a production capacity of 1.5 tonnes a day from CPO.
The new 'crude cartel'
Indonesia is currently the world's second-largest producer of CPO, lagging behind only neighboring Malaysia. Together, the two Southeast Asian countries account for 84% of total world production and 88% of global exports. Significantly, Indonesia and Malaysia are now in talks about the establishment of a cartel-like strategic alliance. Although key issues such as profit-sharing and technology transfer are still under negotiation, once up and running a CPO cartel would potentially give the two countries a large degree of supplier control over the commodity's global price.
That is, if all goes according to plan. CPO production in Indonesia is forecast at 15.2 million tonnes this year, enough perhaps to transform it into Asia's largest bio-diesel producer. Domestic consumption is currently estimated at 30-40% of total production, leaving about 9 million tonnes for biofuel processing and export.
Commodity analysts note that Indonesia's and Malaysia's palm-oil industries differ in important ways that, unless rectified, could result in Malaysia gaining significantly more from the emerging global appetite for biofuels.
While Indonesia has plenty of land and cheap labor, it lacks the capital and technical know-how needed to maximize these advantages. Malaysia, which has a substantially more skilled workforce, has already hit a significant land barrier to palm-plantation expansion and rising labor costs have contributed to stinting new investments.
Nearly 25% of Indonesia's current CPO production is derived from Malaysian investments. Indonesian nationalists, some represented in parliament, have long complained that Indonesia serves as merely a source of raw material and cheap labor for Malaysia's better-funded, more sophisticated and considerably more profitable operations.
Moreover, Malaysia has recently gone on a buying spree of Indonesian oil-palm plantations. Global capital, too, is increasingly bidding to get in on the act: a US-Malaysian joint venture recently announced plans to build a new biofuel-processing plant on Batam, in Indonesia's Riau province, a mere 30 kilometers southeast of Singapore. Riau, after massive deforestation, has emerged as the largest palm-oil-producing area in Indonesia.
How the two sides handle these potentially volatile issues will go a long way in deciding the success or failure of their emerging cartel arrangement. Another big question is whether plantation owners will actually assist the Indonesian government's new drive toward more biofuel production and agree to sell their CPO locally rather than exporting it.
The latter issue cannot be taken for granted. Indonesian CPO producers have recently complained that the government views them merely as another source of state revenue now that global demand for the commodity is surging. Agriculture Minister Anton Apriyantono has notably started to play the nationalist card in appealing to CPO farmers. "For the sake of the national interest, I hope the industry will sacrifice some of its profits in order to develop the bio-diesel industry," he recently said.
Indonesian Palm Oil Producers Association (GAPKI) chairman Derom Bangun said its members would not be willing to sacrifice without considerably more government support for the industry. He said the government should "learn from Malaysia", where CPO producers receive a healthy subsidy for selling their oil to the bio-diesel industry.
Agustino Sudjono, a senior executive at the publicly listed CPO producer London Sumatra Indonesia (Lonsum), notes that while Malaysia is expected to have a production capacity of 1.2 million tonnes of bio-diesel by 2007, which will inevitably increase its demand for CPO, in comparison, "we are still lagging behind and waiting for the regulation to be made official".
CPO: Environmental friend or foe?
So, is this the dawn of a new biofuel-driven era of economic prosperity in Indonesia? There are still many unanswered questions concerning the long-term sustainability of biofuels, chief among them the future price of oil. While reducing dependence on fossil fuels is now globally en vogue because of sky-high pump prices, it could be a passing trend if petroleum prices eventually return somewhere close to their historical moving average.
The adverse impact of palm plantations on Indonesia's and the broader region's natural environment is also a potential deal-breaker. A recent report by the environmental group Friends of the Earth claims that the development of oil-palm plantations was responsible for an estimated 87% of all deforestation in Malaysia. Forest fires in Indonesia, which sometimes cover neighboring countries in smog, are started mainly by palm growers to clear land for new planting.
There is a social cost to plantation expansion, too. In Indonesia, palm-plantation owners frequently clash with local residents over land-ownership rights, while aggrieved locals often protest violently over what they perceive to be only half-hearted corporate-led community development programs. Thousands of indigenous people have been evicted from their lands in recent years, with some beaten and even tortured when they resist, according to rights groups' accounts. This has predictably led to widespread local opposition to the establishment and expansion of plantations.
Environmentalists have already drawn blood over Jakarta's plan, funded by China, to create the world's largest integrated oil-palm plantation, including processing facilities, which would run along the mountainous 850km border with Malaysia in Kalimantan. The environmental group WWF warns that the plan would have a devastating impact on the forests, wildlife, and indigenous community. Most of the mountainous region, known as the "Heart of Borneo", is covered in prime forests and is considered one of the richest areas of biodiversity on Earth.
Minister Apriyantono announced last week that only 180,000 hectares, or 10% of the planned 1.8 million hectares of rainforests, would be used for palm-oil development. However, law enforcement is still dicey in Indonesia's hinterlands. As the country gears up for more biofuel production - before the industry has even taken root - hard questions about long-term sustainability are certain to crop up.
Bill Guerin, a Jakarta correspondent for Asia Times Online since 2000, has worked in Indonesia for 20 years, mostly in journalism and editorial positions. He has been published by the BBC on East Timor and specializes in business/economic and political analysis related to Indonesia. He can be reached at softsell@prima.net.id.


Our supporter Pippa Stacey has released a compilation book featuring multiple contributors sharing their experiences of varying severities of M.E.
The book, titled Dear Chronic Illness, features a collection of short letters written by young people, addressed to their long-term illnesses and impairments, detailing what they would like to say to them.
It's a short read, comprising of 61 pages, and will be released in eBook and paperback format from Amazon.
Royalties from the book will be donated to Spoonie Survival Kits, which Pippa founded and which provides "little bags of happiness that contain a range of items that will hopefully help those with long-term conditions to smile, and remind them they are not alone."
Pippa said: "I'm absolutely thrilled for the release of Dear Chronic Illness. It's such a privilege to have this platform and to be able to share stories of what it's really like to be a young adult with a long-term illness: we hope that in doing so, it will help others to feel less alone and encourage them to share their own story as well."
"I can't say thank you enough to Leesa Wallace and the team at Wallace Publishing for their invaluable support and encouragement, and for helping us to sustain the running of Spoonie Survival Kits too."
You can find out more about the book and how it came about on the Spoonie Survival Kits' website.


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Home/News/School intensifies disaster preparedness
School intensifies disaster preparedness
"Some time last year we had two terrible disasters. One of our grade one learners was knocked down by a car on her way to school, while crossing the busy Beitbridge to Gwanda road. Fortunately, the girl had minor injuries and was rushed to hospital and she survived," says Edson Ncengani the Headmaster at Malala Primary School in Beitbridge District.
"In February last year, our school administration block and the adjacent classroom block were struck by lightning. Luckily the incident happened when the learners had been dismissed from school. There was severe damage to students' exercise books, furniture and some stationery that was inside the affected classroom," Ncengani said.
These two incidents were a wake-up call for the school authorities at Malala. One year later, Malala Primary School which enrols 1017 children from ECD to Grade 7, has intensified efforts to prepare and respond to any disaster that may strike. This is being done to reduce the risk of losing lives among school children and teachers, as well as loss to school property. The Malala Primary School disaster risk reduction initiative is being implemented in partnership with Save the Children, with funding from the European Union Humanitarian Aid.
"All the teachers at our school received some training on disaster risk management. Four of our teachers attended high level workshops in the district, and came back to conduct disaster preparedness sessions with all the learners from ECD to grade 7," added Ncengani.
"Learners can now single out possible disasters, how to prevent them and how to react whenever such disasters occur. We acknowledge that children by their nature may forget or overlook some stuff, so we continue to conduct refresher sessions for the children on disaster risk reduction (DRR)," said the headmaster.
Ncengani said the school has made arrangements with parents and teachers to help the learners in the infant grades to safely cross the road.
"Each morning, the parents accompany the learners to school, and help them to cross the road. When the infant learners dismiss, we make it a point that every day the teachers take turns to assist the children in safely crossing the busy Beitbridge to Bulawayo highway, which is situated just outside our school's main gate," he said.
"This road is really a death trap, so we make sure that the teachers and parents assist the learners to cross to and from school. During the rainy season, the parents also take turns to help the children to cross the nearby river safely on their way to school."
Once in a while, the teachers and learners from the school participate in practical sessions on disaster risk reduction, so as to raise awareness on possible disasters and help the children to better manage the risks. These include visits to the highway to learn about how to safely cross the road, visits to nearby rivers and other possible disaster areas for learning purposes.
In addition, two of the teachers at the school, Terezia Hwande and Tryphine Moyo have been appointed the school's Disaster Risk Reduction focal persons. They have since trained both teachers and learners in conducting practical sessions on DRR that include fire drills.
"In case of an emergency we blow our whistle as a way of communicating with the children and staff, and they move quickly in a single file and go to our disaster risk management assembly point. We then do a roll call to check if all the children are safe and to tell them the way forward," said Terezia Hwande.
This is done to avoid stampedes, which may result in further loss of lives, as well as accounting for all the students. These efforts are helping the school to remain prepared to manage any disaster.
Malala Primary School, is among the 60 schools selected from Beitbridge and Matobo Districts, which are participating in the School Based Disaster Risk Reduction initiative. This is part of the Ready to Act project being implemented by Save the Children, OXFAM and International Rescue Committee with funding from the European Union Humanitarian Aid.
DRR & CCA
Save the Children International is a company limited by guarantee, registered in England and Wales with company number 3732267 and a charity registered in England and Wales with charity number 1076822. Its registered office is St Vincent House, 30 Orange Street, London, WC2H 7HH.
© 2019 Save the Children International | Legal information


RENO, NV; January 22ND, 2009 – The Mexico Tourism Board and the Safari Club International, the leader in protecting the freedom to hunt and in promoting wildlife conservation worldwide is teamed up during the annual Safari Club International's annual convention on January 21 – 24, to promote Mexico's popular hunting destinations.
Destinations such as including Zacatecas, Aguas Calientes, Tamaulipas, Coahuila, Baja California Norte and Durango joined in the inauguration of the Mexico pavilion where a ceremonial ribbon cutting took place.
Mexico ranks 4th among the amount of mammal species and 11th in bird species. For those looking to hunt deer, there are 14 subspecies of white-tailed deer, 6 subspecies of mule deer and 2 species of tamazate deer from the tropical forests. For the Mexican Turkey Slam, you can find pavo ocelado in the tropical forests of Campeche and Gould's and Rio Grande turkey in central Mexico.
SCI fights tirelessly to protect the hunting heritage enjoyed by 15 million Americans and 45 million families around the globe. SCI Members are active in more than 100 countries. SCI's Convention is the most dynamic and diverse hunting show today. It is the most powerful buying and selling environment in the outdoor industry.
I thought that Mexico was second in tne number of mammal species. I urge this industry to ban hunting on Elegant Quail and other endemic species. There numbers are down.


Glucosamine is naturally produced by the body to protect the ends of bones, like shoelace-caps. They are usually derived from crustaceans.
osteoarthritis and reduces breakdown of cartilage.
and curcumin for optimizing joint health.
component of the connective tissue in the skin.
to mankind. It is used to treat tumours in animals with resounding success.
of the cartilage building blocks will be available for cartilage repair.
Glucosamine is a part of a molecule called a glycosaminoglycan-this molecule is used in the formation and repair of cartilage.
formation of new cartilage by providing more of the necessary building blocks.
like shoelace-caps. They are usually derived from crustaceans.


Polls open on Election Day 7 a.m. to 7 p.m.
Polls on Election Day will be open from 7 a.m. to 7 p.m.
MURFREESBORO – The Rutherford County Election Commission will serve voters from 7 a.m. to 7 p.m. today with the help of about 350 Election Day poll workers.
Early voting for today's election concluded Thursday night with a final count of 27,823 out of 155,795 registered voters participating. The last two days of early voting had a strong showing, and Farley foresees that to be the case on Election Day.
The ballots in Murfreesboro and Smyrna also include referendums on whether grocery stores should be permitted to sell wine. All of the ballots have four amendments to the Tennessee Constitution, including Amendment No. 1 to give state lawmakers authority to set limits and regulations on abortions.
Voters can speed up the process by researching the ballot in advance, especially on the amendments, election commission staff recommend.
Each of the 48 precinct polling places will have ballots posted on walls, and smaller paper ones will be available for voters to examine while they wait in line, Farley said.
Voters also need to bring an acceptable form of photo identification, such as a Tennessee drivers license or a military ID card or Veterans Affairs card, Farley said.
State- and federal-government issued photo ID cards are acceptable unless they are student ID cards, Farley said.
Voters who forget to bring their ID cards can still cast provisional paper ballots that will be counted as long as they show up within two days during business hours to present their acceptable form of photo ID to the election commission staff on the south side of the Public Square in Murfreesboro, Farley said.
People can check their voter registration cards to see the location and address of their polling places, and the cards identify the districts for the voters.
The voters who have lost registration cards can find out their polling information, candidates, districts and elected officials through downloading an app through a state website: www.GoVoteTN.com, Farley said.
Those unable to find out their polling location through the Internet can call the election commission office at 615-898-7743.
In addition to offering 48 polling places, the election commission has two "fail safe" voting centers for those who changed their county address without notifying the election commission. Voters who recently moved should go the fail safe centers, Farley said, either at the election commission's Annex Building at 426 E. Vine St. in Murfreesboro or at the Smyrna Fire Administration Headquarters at 145 S. Lowry St.
The total early voting count includes 11,773 votes cast in the Murfreesboro City Council election. Another 4,622 voted in the Smyrna election for mayor and Town Council candidates, and 2,639 voted in the La Vergne Board of Mayor and Aldermen election.


text:
Alexander Meigs "Al" Haig, Jr. (Filadelfia, Pensilvania; 2 de diciembre de 1924-Baltimore, Maryland; 20 de febrero de 2010) fue un militar y político estadounidense. General retirado, fue Jefe de Gabinete de la Casa Blanca entre 1973 y 1974, durante el Escándalo de Watergate, y luego fue Secretario de Estado entre 1981 y 1982.

Biografía

Primeros años

Nació en una familia católica de Filadelfia, Pensilvania. Tras asistir a colegios católicos de la ciudad, ingresó en la Universidad de Notre Dame por un año, antes de trasladarse a la Academia militar de West Point, donde se graduaría en 1947. Entre 1954 y 1955 estudió en la Escuela de Negocios de la Universidad de Columbia, y en 1961 realizó un máster en Relaciones Internacionales en la Universidad de Georgetown, con una tesis sobre el papel del ejército en la política nacional.

Héroe de guerra

Asignado como asesor del General Douglas MacArthur durante la ocupación de Japón, sirvió a sus órdenes en la Guerra de Corea (1950-1951). Haig fue el encargado de mantener el mapa de situación, y de ofrecer al General MacArthur una sesión informativa diaria sobre los eventos en el campo de batalla. En Corea, participó en siete campañas militares, entre ellas la Batalla de Incheon, la Batalla de la Reserva de Chosin y la Evacuación de Hungnam. Fue condecorado con dos Estrellas de Plata al heroísmo y una Estrella de Bronce.

Fue destinado a Europa en los años 50, y en 1960 volvió a los Estados Unidos como profesor en la Academia militar de West Point y oficial del Departamento de Defensa. Trabajó en la oficina del adjunto al Jefe de Personal de Operaciones en el Pentágono (1962-1964), como asesor militar del Secretario de la Marina Stephen Ailes (1964) y asistente del Secretario de Defensa Robert McNamara (1964-1966).

Volvió al campo de batalla en 1966, como comandante de un batallón de la 1.ª División de Infantería en la Guerra de Vietnam. En marzo y abril de 1967, en Ap Gu, a 110 kilómetros de Saigón, el 1.º batallón del 26.º Regimiento de Infantería de la 1.ª División, dirigido por Haig, libró la última batalla importante de la segunda fase de la Operación Junction City contra el Vietcong. Cuando sus tropas quedaron inmovilizadas por fuerzas del Vietcong que los triplicaban en número, Haig, en contra de las recomendaciones, montó en helicóptero para volar hasta el punto de contacto y tener una visión general del campo de batalla. El helicóptero fue derribado, pero el coraje y determinación mostrado por Haig inspiró a sus hombres en dos días de lucha cuerpo a cuerpo con el Vietcong. A pesar de la inferioridad numérica, los hombres de Haig fueron capaces de dar muerte a 592 soldados vietnamitas.

Promovido al rango de Coronel, Haig fue condecorado por el General William Westmoreland con la Cruz al Servicio Distinguido, la segunda condecoración más importante al heroísmo. También recibió la Cruz Distinguida de Vuelo y el Corazón Púrpura.

Protegido de Henry Kissinger

A su vuelta a los Estados Unidos, se incorporó como asistente militar al equipo del Consejero de Seguridad Nacional Henry Kissinger, en la nueva Administración Nixon (1969-1970). En 1970, el Presidente Nixon lo promovió para Adjunto al Consejero de Seguridad Nacional (1970-1973). En esa responsabilidad defendió bombardeos masivos contra Camboya y Vietnam del Norte, y ayudó al presidente Nguyen Van Thieu, de Vietnam del Sur, a negociar el alto el fuego en 1972. También pidió al FBI que pinchara los teléfonos de catorce funcionarios del Gobierno y tres periodistas para averiguar quién estaba filtrando información reservada a los medios.

En 1973 fue nombrado vicejefe del Estado Mayor del Ejército.

Jefe de Gabinete de la Casa Blanca (1973-1974)

El 30 de abril de 1973, H.R. Haldeman dimitió como Jefe de Gabinete del Presidente Nixon, y Haig fue reclamado para cubrir la vacante. Le tocó dirigir el Gabinete presidencial en los quince últimos meses de mandato de Nixon (mayo de 1973 - agosto de 1974). Se ganó la reputación de buen administrador en el manejo de los asuntos del día a día en medio de la crisis provocada por las investigaciones sobre el escándalo Watergate. También se dedicó a vigilar al Presidente en momentos de crisis personal. Solo él y otras dos personas, Rosemary Woods, secretaria personal del Presidente, y Stephen Bull, asistente del Presidente, tuvieron conocimiento de la supresión de algunas partes de las cintas entregadas a la justicia, de las grabaciones efectuadas en el Despacho Oval.

Si bien en un primer momento aconsejó a Nixon que resistiera, más tarde fue uno de los que le convencieron de la necesidad de dimitir. Dirigió la transición presidencial en los primeros días de mandato de Gerald Ford, y jugó un papel importante para que el nuevo Presidente otorgara el perdón a Nixon.

Comandante Supremo de la OTAN (1974-1979)

El 1 de noviembre de 1974, el Presidente Ford lo nombró Comandante supremo aliado en Europa de la Organización del Tratado del Atlántico Norte (OTAN). En esa responsabilidad defendió el papel de la OTAN como garantía contra los procesos revolucionarios en Europa. Frente los aires revolucionarios en Portugal, la Alianza participó en un movimiento de presión para hacer volver las cosas a la normalidad; además de las presiones del Fondo Monetario Internacional, se vio la presión de la OTAN, para volver atrás el proceso revolucionario portugués. En Italia advirtió contra la posible injerencia de los comunistas en ministerios claves como Defensa, Interior y Asuntos Exteriores. Y en España abogó por la estabilización del proceso democrático interno, y el pronto alineamiento del país en la OTAN.

El 25 de junio de 1979, escapó de un intento de asesinato en Bélgica por parte de la Facción del Ejército Rojo. Los terroristas alemanes colocaron una mina terrestre al paso de la comitiva del General Haig, hirieron a tres de sus guardaespaldas que viajaban en otro coche.

Ese mismo año, Haig se retiró definitivamente del ejército, y pasó a ocupar la presidencia de United Tecnologies, sociedad vinculada a la fabricación de material militar.

Secretario de Estado (1981-1982)

En sus años como Comandante Supremo de la OTAN, se había ganado la atención de los sectores políticos conservadores en Estados Unidos, gracias a su oposición a la decisión del presidente Jimmy Carter de cancelar la bomba de neutrones, y sus reservas sobre el tratado SALT II. Y a finales de 1980 fue recomendado al Presidente electo Ronald Reagan para ocupar el cargo de Secretario de Estado. Tras un tumultuoso proceso de confirmación en el Senado, en el que se puso en cuestión su papel durante la Administración Nixon, fue confirmado gracias a su condición de héroe de guerra con 93 votos a favor y 6 en contra. Se convirtió en el segundo militar de carrera en ocupar ese puesto —el otro había sido George Marshall.

Entre sus prioridades estaría hacer frente al expansionismo soviético, con especial énfasis en América Central, donde consideraba que el poder soviético se había transformado de defensivo a ofensivo. Trazó una línea estratégica en el Canal de Panamá para fijar los límites de la zona de seguridad de Estados Unidos. Dentro de esa zona se incluían Nicaragua, Guatemala, y El Salvador, países de interés supremo para Estados Unidos. Para Haig, el principal peligro para la Seguridad Nacional era que el límite de seguridad llegara hasta la frontera con México. Impulsó las primeras acciones encubiertas contra las guerrillas de Guatemala, el Frente Farabundo Martí de El Salvador, y el Gobierno sandinista de Nicaragua; y el fortalecimiento del cuerpo diplomático norteamericano en la vecina Honduras, un aliado.

Aunque los aliados apreciaron su designación por su conocimiento de los problemas europeos, pronto desconfiaron de sus modales ariscos y destemplados. Una de sus primeras decisiones como Secretario de Estado fue prohibir al embajador soviético Anatoli Dobrynin, decano del cuerpo diplomático de Washington, estacionar su coche en el aparcamiento del Departamento de Estado, en un espacio privilegiado que un día le concediera por cortesía Henry Kissinger. Y en noviembre de 1981, indignó a los aliados europeos al advertir que Estados Unidos tenía previsto en sus planes utilizar una bomba atómica en Europa en caso de una agresión convencional soviética.

Su estrategia agresiva logró algunos éxitos, como la paralización de la diplomacia soviética en los últimos meses del gobierno de Leonid Brézhnev, o cierto acercamiento a los partidos socialdemócratas europeos, destacando su apoyo a la alternativa ganadora de Felipe González en España. Pero lo condenaron sus malas relaciones con otros departamentos del Gobierno. Trató de introducir una completa red de ayudantes en el Departamento de Estado y en otros organismos de la Administración. El Consejo de Seguridad Nacional y el Pentágono sufrieron sus continuas intrigas para ampliar su red de poder. Pero nunca logró ganarse la confianza del Presidente Reagan, ni acceder a su círculo personal.

Para su segundo año como Secretario de Estado, ya había creado suficientes resistencias en el servicio diplomático y antipatías extranjeras como para hacer recomendable su alejamiento. Dimitió en julio de 1982.

Atentado contra Reagan (30 de marzo de 1981)

Mientras el presidente Ronald Reagan era operado en el Hospital George Washington después de haber recibido un balazo, Haig fue el encargado de informar de lo ocurrido a los Gobiernos aliados y a la Unión Soviética. Y ante la ausencia del vicepresidente George Bush, que se encontraba de viaje en Texas, Haig tomó el mando en la reunión de urgencia del Gabinete en la Sala de Situaciones de la Casa Blanca.

El subsecretario de prensa Larry Speakes fue preguntado por los periodistas sobre quién estaba al mando del Gobierno. "No puedo responder a esa pregunta en este momento", fue su respuesta. Tras escuchar a Speakes, Haig irrumpió en la Sala de Prensa de la Casa Blanca para decir: "Estoy al mando en este momento. Constitucionalmente, caballeros, tenemos al Presidente, al Vicepresidente y al Secretario de Estado, en ese orden. Si el presidente decidiera que quiere traspasar los poderes al Vicepresidente, lo podrá hacer. Mientras tanto, yo estoy al mando aquí, en la Casa Blanca, pendiente de la vuelta del Vicepresidente y en contacto con él."

Lejos de tranquilizar a la población, sus palabras fueron interpretadas como un exceso en su autoridad. En sus palabras, Haig se saltó la Constitución. El tercero en la línea de sucesión es el Presidente de la Cámara de Representantes (Speaker of the House) y el cuarto el Presidente Pro tempore (el senador de más antigüedad en el senado), quedando de quinto el Secretario de Estado. Más tarde se defendió asegurando que no hablaba en términos legales.

Guerra de las Malvinas (1982)

Tras la ocupación de las islas Malvinas por parte de la dictadura argentina, y en contra de la opinión de otros elementos de la Administración Reagan, Alexander Haig creyó que Estados Unidos debía desempeñar un papel de mediador, para evitar que América Latina reaccionara contra Estados Unidos si se ponían de forma automática del lado británico. El 8 de abril de 1982, inició una intensa gira diplomática de varias escalas por Londres y Buenos Aires.

Su primer planteamiento se basó en intentar persuadir a las dos partes —Argentina y Reino Unido— de aceptar algún tipo de administración provisional neutral, con posterioridad a la retirada de Argentina, que se hiciera cargo de la administración de las islas mientras se decidía su futuro a largo plazo. Sus propuestas incluían la creación de una comisión, en sustitución del Gobernador, compuesta por representantes de Estados Unidos, Reino Unido y Argentina, que actuarían de forma conjunta para asegurar el cumplimiento del acuerdo de paz que se alcanzara, y las negociaciones en cuanto a una solución duradera se desarrollarían en función de los objetivos y principios de la Carta de Naciones Unidas.

Pero los argentinos se resistían a la necesidad del retiro de sus tropas de las Malvinas, tal y como señalaba la resolución 502 del Consejo de Seguridad de las Naciones Unidas, y los británicos mantenían que la soberanía británica debía prevalecer, y la administración británica debía ser restaurada. Los esfuerzos de Haig fracasaron y el 19 de abril volvió a Washington.

En diciembre de 2012 documentos desclasificados demostraron que Haig planeaba revelar información militar clasificada británica a la Argentina antes de la Operación Paraquet. La propuesta, que habría revelado los planes británicos para la recuperación de la isla, tenía la intención de mostrar a la junta militar en Buenos Aires que Estados Unidos era un actor neutral y se podía confiar en que actuara imparcialmente durante las negociaciones para poner fin al conflicto.

Últimos años

John Dean, exconsejero del presidente Richard Nixon, publicó en 1982 un libro en el que señalaba a Alexander Haig como la fuente secreta del Washington Post en el escándalo Watergate. Haig desmintió las acusaciones y las describió como debidas a motivaciones comerciales de Dean. En 2005 se confirmó que la identidad de Garganta Profunda correspondía en realidad a Mark Felt, antiguo número dos del FBI.

En 1987 presentó su candidatura a la nominación del Partido Republicano para las elecciones presidenciales de 1988. Cuando abandonó la carrera por falta de apoyos, prestó su apoyo al Senador Bob Dole, de Kansas, y criticó duramente al vicepresidente George Bush. Siempre pensó que Bush tuvo parte de responsabilidad en su rápida salida del Departamento de Estado, porque lo consideraba un futuro rival.

Falleció el 20 de febrero de 2010 a la edad de 85 años.

Referencias

Enlaces externos

Políticos de Estados Unidos
Secretarios de Estado de Estados Unidos
Generales de Estados Unidos
Comandantes supremos aliados en Europa
Condecorados con el Corazón Púrpura
Condecorados con la Estrella de Bronce
Alumnado de la Universidad de Notre Dame
Caballeros de la Orden de Malta
Condecorados con la Estrella de Plata
Doctores honoris causa por la Universidad de Haifa
Jefes de Gabinete de la Casa Blanca
Diplomáticos de la guerra de las Malvinas
Nacidos en Filadelfia
Fallecidos en Baltimore
Católicos de Estados Unidos
Republicanos de Maryland
Republicanos de Pensilvania

redpajama_set_name: RedPajamaWikipedia


Adria Grotesk font is a family of 7 weights designed by Facetype / Marcus Sterz. We were very excited to find how beautiful Adria is – a sans-serif family with perfect smooth outlines, large openings and excellent readability – all these touched by excellent stylish humanist terminals. Another great surprise were the upright italics – one for each weight sold as separate fonts. Designed with same perfection as regulars, italics have more vintage look flavored with the style of natural handwriting. Adria is developed with really lots of open type features and supports full list of multilingual diacritics, a lot of options for numbers, alternates, swashes and ligatures. It is a perfect choice for strong headlines, subheads and large paragraphs of small body text. It would look excellent on your package, book cover, typographic poster, stationary or logo.


A proposed final scene for Avengers 4, which reunites Steve Rogers and Peggy Carter for one last dance, has made Marvel fans on the internet collectively weep. Read it here.
The idea takes off from Steve's tragic romance with Peggy. At the end of Captain America: The First Avenger, Steve sacrifices himself to save the lives of millions. He deliberately crashes a plane that's carrying bombs, but all the while he's talking to Peggy on the radio. "This is my choice," he says, as he puts her picture on the dials in front of him and plunges the plane down. "I'm going to need a raincheck on that dance," he says, and she sets a date, with tears in her eyes, knowing that he'd never make it.
Aside from a brief meeting in Captain America: The Winter Soldier and a nightmare scene in Avengers: Age of Ultron (which once again brought up the unrealised dance), Peggy and Steve next meet as she is on her deathbed in Captain America: Civil War, after which a new romantic plot between Steve and Peggy's niece was introduced and quickly abandoned.
While nothing is confirmed yet, it has been rumoured that Steve Rogers, along with many others, will die at the end of Avengers 4, the film that will essentially wipe the slate clean for a fresh phase of Marvel films. It has also been rumoured that Hayley Atwell, who plays Peggy Carter in the MCU, filmed scenes for the fourth Avengers film when pictures of hers wearing motion capture dots on her face were shared online.
Chris Evans' contract with Marvel is said to be ending after Avengers 4, after which the actor will likely hand over the baton to a younger bunch of actors. In Avengers 4, the surviving superheroes will attempt to correct the events of Avengers: Infinity War, and save the 'dead' Avengers.


The Fisher Community Fair and Horse Show began in 1941 as a grouping of tents behind the old Fisher Elementary School. Because of gas and rubber rationing during World War II, the residents of the area were not able to travel to the Champaign County Fair. Therefore, the residents of the Fisher area initiated a community fair in which the agricultural community could exhibit its hard work. You can contact Jill Hardesty for more information about the fair.


text:
Tony Igele
We Are Ready To Re-Orientate Our People – Obaseki
Edo State Governor Godwin Obaseki said his administration is ready to re-orientate the citizens of the state and also entrench trust between the government and the people.
He made this known when he received Executive Members of the National Orientation Agency (NOA), Edo State Directorate led by the Acting State Director of the Agency Mrs. Grace Ewere Eseka on a courtesy visit to government house in Benin City Edo State.
Obaseki said there is nothing new about relating and interacting with the people but rather the emphases should be on entrenching trust between the government and the Edo people.
"When we design our programmes and leave them at the level we are today without really domesticating and localising them in such a way that people can relate directly with them, then we will be missing out the point".
He said the issue is beyond sloganeering and nice songs on television but rather strive to find a way to affect the people on a daily bases. "We want to move beyond sloganeering. We have the Ministry of Orientation and Communication because we believe in the progressive steps we are taking to get our people to think, how we lead and run our society"
He said his administration has decided to go beyond talking and acting but rather make the people to see government action and create room for them to judgement government action.
"In every aspect of life there has to be a change and orientation for the society to work. So it is not about restructuring at the top but it is about restructuring at the base."
The governor said change should begin with us as individuals saying it is time for hard work. "Change begins with me but how, what do we do everyday to make it happens? This is the time for hard work and that is what we want you to do with us".
Earlier, the Acting State Director of the National Orientation Agency Mrs. Grace Ewere Eseka said they are in government house to intimate the governor of the programme lined up for implementation of the Agency and seek for the state government's support in facilitating the adoption and delivery of the programmes for the over all benefits of the people.
She said NOA is charge to work with State Ministry of Communication and Orientation, to actualize these progressive initiatives for the benefit of the people. "The task is onerous, but worth working together for, to ensure that government engages the people through meaningful and consistent dialogue, as it is appropriate. This will go a long way in bridging the information-gap between the government and the public".
Mrs. Eseka said the Agency is working hard at ensuring that the people come closer to their elected officials through mutual and inclusive confidence-building, openness, and improved public-government relationship.