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|Immunoglobulin heavy chain variable region genes contribute to the induction of thyroid-stimulating antibodies in recombinant inbred mice.|
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|PMID: 20407472 Owner: NLM Status: MEDLINE|
|Graves' hyperthyroidism is an autoimmune disease occurring spontaneously in humans and caused by autoantibodies that stimulate the thyrotropin receptor. In mice, inducing Graves'-like hyperthyroidism requires in vivo expression of the thyrotropin receptor using plasmid or adenovirus vectors. However, mice with different genetic backgrounds vary markedly in their susceptibility to induced hyperthyroidism. Further, in some strains major disparities exist between the induction of hyperthyroidism and detection of thyroid-stimulating antibodies. To break tolerance, virtually all Graves' mouse models involve immunization with human thyrotropin-receptor DNA and the standard thyroid-stimulating antibody bioassay uses cells expressing the human thyrotropin receptor. We hypothesized, and now report, that disparities between hyperthyroidism and thyroid-stimulating antibody bioactivity are explained, at least in part, by differential antibody recognition of the human vs the mouse thyrotropin receptor. The genetic basis for these species differences was explored using genotyped, recombinant-inbred mouse strains. We report that loci in the immunoglobulin heavy chain variable region as well as in the major histocompatibility complex region contribute in a strain-specific manner to the development of antibodies specific for the human or the mouse thyrotropin receptor. The novel finding of a role for immunoglobulin heavy chain variable region gene involvement in thyroid-stimulating antibody epitopic specificity provides potential insight into genetic susceptibility in human Graves' disease.|
|B Rapoport; R W Williams; C-R Chen; S M McLachlan|
|Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't|
|Title: Genes and immunity Volume: 11 ISSN: 1476-5470 ISO Abbreviation: Genes Immun. Publication Date: 2010 Apr|
|Created Date: 2010-04-21 Completed Date: 2010-08-20 Revised Date: 2014-08-27|
Medline Journal Info:
|Nlm Unique ID: 100953417 Medline TA: Genes Immun Country: England|
|Languages: eng Pagination: 254-63 Citation Subset: IM|
|APA/MLA Format Download EndNote Download BibTex|
Genes, Immunoglobulin Heavy Chain / genetics*
Genome-Wide Association Study
Graves Disease / genetics, immunology
Hyperthyroidism / genetics, immunology
Immunization / methods
Immunoglobulin Variable Region / genetics*
Immunoglobulins, Thyroid-Stimulating / blood, genetics, immunology*
Mice, Inbred C3H
Mice, Inbred Strains
Receptors, Thyrotropin / genetics, immunology
|DK19289/DK/NIDDK NIH HHS; DK54684/DK/NIDDK NIH HHS; P20-DA-21131/DA/NIDA NIH HHS; R01 DK054684/DK/NIDDK NIH HHS; U01AA13499/AA/NIAAA NIH HHS; U01CA105417/CA/NCI NIH HHS; U24AA13513/AA/NIAAA NIH HHS; U24RR021760/RR/NCRR NIH HHS|
|0/Immunoglobulin Variable Region; 0/Immunoglobulins, Thyroid-Stimulating; 0/Receptors, Thyrotropin|
Journal ID (nlm-journal-id): 100953417
Journal ID (pubmed-jr-id): 21482
Journal ID (nlm-ta): Genes Immun
Journal ID (iso-abbrev): Genes Immun.
nihms-submitted publication date: Day: 11 Month: 6 Year: 2014
Print publication date: Month: 4 Year: 2010
pmc-release publication date: Day: 23 Month: 7 Year: 2014
Volume: 11 Issue: 3
First Page: 254 Last Page: 263
PubMed Id: 20407472
|IMMUNOGLOBULIN HEAVY CHAIN VARIABLE REGION GENES CONTRIBUTE TO THE INDUCTION OF THYROID STIMULATING ANTIBODIES IN RECOMBINANT INBRED MICE|
|Basil Rapoport, MD1|
|Robert W. Williams, PhD2|
|Chun-Rong Chen, MD1|
|Sandra M. McLachlan, PhD1|
1Autoimmune Disease Unit, Cedars-Sinai Research Institute and UCLA School of Medicine, Los Angeles, CA
2Department of Anatomy and Neurobiology, University of Tennessee Health Science Center, Memphis TN
|Correspondence: Corresponding Author: Sandra M. McLachlan, Autoimmune Disease Unit, Cedars-Sinai Research Institute. 8700 Beverly Blvd B-131, Los Angeles 90048, CA; email@example.com; Telephone: 310-423-7680, Fax: 310-423-0221
The thyroid is the organ most commonly affected in human autoimmunity.Hashimoto’s thyroiditis is associated with autoantibodies to thyroid peroxidase and thyroglobulin and hypothyroidism affects 4.6 % of the US population 1. Hyperthyroidism in Graves’ disease, prevalent in 0.5 - 1% of women 1,2, is caused by autoantibodies to the thyrotropin receptor (TSHR) that mimic thyroid stimulation by thyrotropin (TSH) (reviewed in 3). Autoimmune hyperthyroidism only develops spontaneously in humans. Nevertheless, in other autoimmune disease, induced animal models have provided important clues to elucidating the pathogenesis and genetic basis of their human counterparts, such as experimental autoimmune encephalomyelitis 4 (a model for multiple sclerosis) and collagen-induced arthritis 5.
Induction of hyperthyroidism in mice is not as straightforward as the two foregoing induced autoimmune diseases. Even though highly-purified, disease-specific antigen (unequivocally the TSHR) is available, conventional immunization with TSHR protein and adjuvant cannot induce hyperthyroidism . Although this approach generates high titers of TSHR antibodies, these are incapable of activating the TSHR on the surface of intact cells (reviewed in 6). To produce Graves’ hyperthyroidism in mice, it is necessary to express in vivo the TSHR or it’s A-subunit using plasmid or adenovirus vectors (reviewed in 6). However, mice with different genetic backgrounds vary markedly in their susceptibility to induction of hyperthyroidism.
Virtually all mouse models of induced hyperthyroidism involve in vivo expression of the TSHR without additional adjuvant (for example 7,8). Human TSHR cDNA is generally employed, because of its general availability and because its use bypasses the need to overcome self tolerance with potent adjuvants. [In one study, hyperthyroidism was induced by injecting B cells expressing mouse-TSHR with the adjuvant cholera toxin B 9]. Importantly, following immunization with the human TSHR, the induced thyroid stimulating antibodies (TSAb) must stimulate the mouse TSHR to produce hyperthyroidism in vivo.
It has been recognized for fifty years that TSAb in human Graves’ patients stimulate the mouse (m) TSHR in vivo10 and early bioassays for TSAb utilized a rat thyroid cell line (FRTL5 11). Following the molecular cloning of the TSHR, Chinese hamster ovary (CHO) cells expressing the recombinant human (h) TSHR have progressively supplanted rat thyroid cells in TSAb assays 12. In BALB/c mice made hyperthyroid by immunization with human TSHR-adenovirus, serum thyroxine levels correlated with TSAb activity when measured with FRTL5 rat thyroid cells 7. Unexpectedly, with the human hTSHR bioassay, TSAb activities in hyperthyroid mice correlated poorly with serum thyroxine levels. For example, some euthyroid BALB/c mice had high TSAb activity while other hyperthyroid animals were TSAb negative 13. Similarly, some C3H/He mice were hyperthyroid despite very low levels of TSAb assayed using hTSHR expressing CHO cells 14. BALB/c mice are far more susceptible than C57BL/6 mice to hTSHR-adenovirus induced hyperthyroidism 15. Nevertheless, when assayed with hTSHR-CHO cells, TSAb activities were comparable in these two mouse strains 15.
To explore the foregoing inconsistencies in the relationship between the degree of murine hyperthyroidism and TSAb activity, we generated mouse-TSHR expressing CHO cells16. Unlike with hTSHR-CHO cells, when assayed with mTSHR-CHO cells TSAb levels were higher in BALB/c than in C57BL/6 mice, more consistent with the greater susceptibility of the former to induced hyperthyroidism 16. With this background, in the present study we hypothesized that TSAb generated in genetically diverse strains of mice and assayed with hTSHR- and mTSHR-CHO cells as separate traits would provide insight into the genetic basis for variability in TSHR antibody functional activity in different mouse strains. We report that loci in the MHC region as well as in the immunoglobulin heavy chain variable (IgVH) region contribute in a strain-specific manner to the development of antibodies specific for the human- or the mouse TSHR.
Adenoviruses, mouse strains and immunization protocols were previously described 14,17. Briefly, we used adenovirus encoding the human A-subunit (TSHR amino acids 1-289; A-subunit-Ad)8 and null-adenovirus (Con-Ad) 18. Propagation, purification and determination of particle virus number was reported previously 7. Female mice (5-8 weeks) of the following strains were obtained (Jackson Laboratory, Bar Harbor, Maine):- (a) C3H/HeJ and C57BL/6J (parental BXH strains); (b) RI CXB1/ByJ through CXB7/ByJ; CXB8/HiAJ through CXB13/HiAJ; (c) RI BXH2-, 4-, 6 through 11-, 14-, 19- TyJ, BXH20/ KccJ , BXH22/KccJ and B6cC3-1/KccJ. Parental strains are referred to as C3H, B6 (Jackson or Bailey strains, J or By), and RI strains as CXB1, CXB 2 or BXH2, BXH4 etc.
Mice were immunized with A-subunit-Ad (108 particles/injection) on 3 occasions at 3 weekly intervals. Blood was drawn one week after the second injection and mice were euthanized four weeks after the third immunization. Six mice were studied for each CXB or BXH strain (except for CXB5; only 2 animals were available). The number of parental animals immunized with A-subunit-Ad were 10 C3H/J and 10 B6/J mice. Additional parental strain mice were immunized with Con-Ad (108 particles/injection): 5 C3H/J and 5 B6/J. All studies were approved by the Institutional Animal Care and Use Committee and performed with the highest standards of care in a pathogen-free facility. All sera had previously been characterized for thyroxine and TSHR antibodies measured by inhibition of TSH binding (TBI) or ELISA 15,17.
TSAb was assayed by stimulation of cAMP generation in CHO cells expressing the human TSHR 14 and the mouse TSHR 16. TSHR-CHO cells in 96 well plates were incubated (80-90 min, 37°C) with test sera diluted 1:20 in Ham’s F12 containing 10 mM Hepes, pH 7.4 and 1 mM isobutylmethylxanthine. After aspirating the medium, intracellular cAMP was extracted with ethanol, evaporated to dryness and resuspended in Dulbecco’s phosphate buffered saline. Samples (12 μl) were assayed using the LANCE cAMP kit (Perkin Elmer, Boston MA). Sera from CXB mice and their parental strains were analyzed for TSAb using mTSHR and hTSHR bioassays; sera from BXH mice and their parental strains were tested for TSAb using mTSHR-CHO cells (data for hTSHR-CHO cells were previously reported 14.
Human and mouse TSAb levels were expressed as a percentage of cAMP values attained with sera from control adenovirus immunized mice and these values were used for linkage analysis (see below). An additional trait used in linkage analysis is the TSAb ratio for mTSHR:hTSHR (BXH strains), and hTSHR:mTSHR (CXB strains). Because maximum cAMP induction differed in the two bioassays, for visual comparisons TSAb values for individual mice were normalized by assigning 100% to the mean value for the strain with the highest response. as follows:- BXH strains, mouse-TSHR 485% (BXH9) and human-TSHR 1229% (BXHcC3-1); CXB strains:- mouse-TSHR 555 % (CXB6) and human-TSHR 945% (CXB6).
Significant differences between responses in different groups were determined by Mann Whitney rank sum test or, when normally distributed, by Student’s t test. Multiple comparisons were performed using analysis of variance (ANOVA). Tests were performed using SigmaStat (Jandel Scientific Software, San Rafael, CA). Data in bar graphs are shown as mean + SEM.
Putative quantitative trait loci (QTL) involved in TSAb induced by A-subunit-Ad immunization of CXB and BXH strains were mapped using the genotype files for these recombinant inbred strains generated by Williams et al. 19 available at http://www.nervenet.org and embedded in GeneNetwork (www.genenetwork.org). The probability of linkage between our traits and previously mapped genotypes was estimated at ~1 centiMorgan intervals (~2 megabase; Mb) along the entire genome, except for the Y chromosome. To establish criteria for suggestive and significant linkage, a permutation test was performed (1000 permutations at 1-centimorgan intervals) 20. This test compares the peak likelihood ratio statistics (LRS; LRS = LOD x 4.6, where LOD is the logarithm of the odds) obtained for a given data set with the peak LRS score obtained for 1000 random permutations of the same data set.
The primary phenotype data have been entered into the mouse CXB and BXH Phenotype databases in GeneNetwork (www.genenetwork.org) under the trait accession identifiers: CXB 10683 through 101690; and BXH 10147, 10148; 10155 through 10160). These data can be found by searching the CXB or BXH; databases for the name “McLachlan”. In the Results we refer to the identification (GN) specific trait numbers so that readers can verify and extend the data analysis. Because GN interval maps connect directly to the UCSC (University of California Santa Cruz) Genome Browser, it is possible to explore the gene complement of chromosomal intervals together with the QTL profile.
CXB and BXH data were combined (when appropriate) to provide a data set from 26 RI strains sharing one parental strain (B6). As described for BXH and BXD RI mice 21 and CXB and BXH mice 14, we calculated the probability associated with a X2 value equal to:- 2(lnPBXD + lnPCXD) with 4 degrees of freedom, where lnPBXD and lnPCXD are the natural logarithms of the probabilities derived independently for the two RI strains in the same chromosomal interval. For the combined analysis, the Gene Network program only provides a LOD score; for comparison with the LRS scores provided by the standard analysis using this program an LRS value of 4.61 is equivalent to a LOD score of 1.0.
C3H and B6 mice were immunized three times with adenovirus encoding the human TSHR A-subunit. Sera were drawn one week after the second injection and one month after the third injection and were tested for TSAb in bioassays using eukaryotic cells expressing the mouse or the human TSHR. As we observed previously 14, TSAb levels determined using the human TSHR were far lower in C3H than in B6 parental strains after the second A-subunit immunization (Fig. 1A). This observation is paradoxical because C3H mice are more susceptible to hyperthyroidism than B6 mice 14. A possible explanation for this phenomenon was that, despite immunization with the human TSHR A-subunit, the TSAb generated in the C3H mice preferentially recognized the mouse TSHR. This hypothesis was confirmed when the same sera were tested with cells expressing the mouse TSHR. In the latter assay, TSAb activities were comparable in C3H and B6 parental strains (Fig. 1B). This difference for each strain can be expressed as the ratio of TSAb activities determined with the mouse and human TSHR assays (Fig. 1C). Similar data were obtained with the sera obtained at the time of euthanasia, four weeks after the third immunization (Supplementary Information Fig. S1).
Based on the foregoing differences between B6 and C3H mice in TSAb determined in mouse and human TSHR assays, we performed similar assays on 13 recombinant inbred BXH strains derived from the two parental strains. BXH strains had been immunized and tested as described above for the parental B6 and C3H mice. Because maximum stimulation levels differed in the two bioassays (Fig.1) and also because TSAb activities induced in the different strains varied widely, to facilitate comparison the data were normalized to the recombinant strain giving the highest mean TSAb value, expressed as 100% (Methods). Six of the 13 BXH strains (BXH-2,-4,-6, -7, -9 and 19) developed significantly higher mouse-TSAb than human-TSAb activity after two immunizations (Fig 2A).
In previous studies, TSHR A-subunit adenovirus immunization of BALB/c(J) and B6(J) mice generated similar TSAb activities when measured in the standard human TSHR assay 15. Recombinant inbred CXB strains were derived from parental BALB/c and B6 mice of the Bailey strains (By, rather than J). We had previously investigated the 13 CXB recombinant inbred strains 14, but had not assayed TSAb activities in these mice following immunization with TSHR A-subunit adenovirus. On doing so in the present study, we found that, in sera tested one week after the second immunization, most CXB strains had comparable TSAb activities measured in the mouse TSHR and human TSHR assays and only one strain (CXB8) had significantly higher TSAb activity in the assay using the human vs the mouse TSHR assay (Fig. 2B).
For these analyses, we used the raw TSAb data (not the values normalized to 100% for better visualization, as shown in Fig. 2). Putative quantitative trait loci (see Methods) were analyzed for the following traits after the second immunization with human TSHR A-subunit adenovirus:- a) TSAb assayed with human TSHR-expressing cells; b) TSAb assayed with mouse TSHR-expressing cells and; c) the ratio between these two TSAb assay values.
In 13 BXH recombinant inbred strains, the whole genome scan revealed highly significant linkage between the ratio of TSAb values determined using the mouse vs. human TSHR assays and a locus on chromosome (Chr 12)(Fig. 3A). The Linkage Ratio Statistic (LRS) of 20.433 is equivalent to a LOD (logarithm of the odds) score of 4.432 (Table 1). Similarly, whole genome scanning for the 13 CXB strains revealed the strongest linkage between a Chr 12 locus and TSAb assayed with mouse TSHR-expressing cells (Fig. 3B). However, data from the same CXB sera assayed for activity using the human TSHR provided evidence for linkage between human TSHR TSAb and loci on Chr 17 and Chr X linked (Fig. 3C). The linkage data obtained in the present study taken together with our previous findings 14,17 (summarized in Table 1) permit the following general conclusions:-
First, loci on the terminal portion of Chr 12 are linked to the following traits involving TSAb after two immunizations with human TSHR A-subunit adenovirus:-
- TSAb measured in the human-TSHR assay for BXH mice (Fig. 4A) and in the mouse-TSHR assay for CXB mice (Fig. 4A);
- the mouse:human-TSHR TSAb ratio in BXH (Fig. 4B) and the human:mouse-TSHR ratio in CXB mice (Fig. 4B).
Second, loci on other chromosomes were shared by TSAb and non-TSAb traits:-
Combining the data for BXH and CXB strains strengthened the linkage between TSAb values (using either the mouse- or human-TSHR or the ratio between these assays) with loci on Chr 12 (Table 2). This combined analysis provides statistical data as LOD (not LRS) scores. LOD scores of 3.49 to 3.56 were obtained for the Chr 12 region from 109.73 to 118.161 Mb, which is dominated by immunoglobulin H chain genes (Fig. 5). The highest LOD scores (>5.8) were obtained for TSAb ratios linked to Chr 12 loci between 110.306 and 111.967 Mb, a region which includes the gene for type 3 deiodinase (Dio3) 22.
TSAb assays performed on sera one month after the third immunization replicated the foregoing data with sera obtained after the second immunization. However, with greater stimulation to the immune system, in addition to the loci on Chr 12, 17 and X, a number of other loci in BXH and CXB mice were linked to TSAb traits, for example on Chr 9 and 10 (Supplementary Information Table S1). The potential significance of these loci is unknown because the genes therein are unidentified.
Before considering our present findings, it should be emphasized that two assays are used to assess TSHR antibody levels in Graves’ patients and induced animal models of this disease. Clinically, the most standardized and cost-effective assay involves antibody mediated TSH binding inhibition (TBI). This assay utilizes purified human or porcine TSHR. A bioassay for functional thyroid stimulating antibodies (TSAb) is more complex and involves measuring the cAMP response to stimulation in non-thyroidal cells expressing the recombinant human TSHR. Early studies with the animal model of Graves’ disease revealed that background genes in different mouse strains had a major effect on the development of hyperthyroidism. Thus, although TSHR antibodies (measured as TBI) were induced in several mouse strains by immunization with TSHR-expressing adenovirus, BALB/c mice were the most susceptible to developing hyperthyroidism 7,23. C57BL/6 mice, which develop high TSHR antibody levels , rarely become thyrotoxic 7.
Because of this strain difference and the dominance of the BALB/c phenotype in F1 offspring of the BALB/c x B6 cross 15, we studied recombinant inbred CXB mice (derived from these parental strains) for genetic factors that control the variability in TSHR antibodies and hyperthyroidism. Our findings suggested that loci on different chromosomes were responsible for TSHR antibody induction versus thyroid function 17. Extending these studies to BXH mice, derived from C3H/He and B6 parental strains, we confirmed the importance of different gene sets controlling immune responses versus thyroid function. In this second study, in addition to measuring TBI, we assayed functional TSAb. We were puzzled to observe that, despite very low TSAb levels, C3H/He mice and some BXH strains became hyperthyroid 14. Our present data resolve this conundrum. Remarkably, despite genetic immunization with a vector expressing the human TSHR A-subunit, C3H/He mice and some BXH strains develop TSAb that preferentially recognize the mouse TSHR, not the human TSHR used in the conventional bioassay. Of course, hyperthyroidism can only develop by activation of the mouse TSHR in vivo. Better antibody recognition of a related antigen than the immunogen, termed “heteroclicity”, is a well established phenomenon. For example, preferential antibody recognition of bovine or porcine insulin after immunization with human insulin 24 and greater affinity early in the immune response for mouse- than for pigeon- cytochrome c after immunization with the latter antigen 25.
In humans, TSAb cross-reactivity with TSHR from different species was first recognized in the late 1950’s, before the immunoglobulin nature of TSHR antibodies was recognized. A component of Graves’ serum was found to stimulate the guinea pig thyroid with a prolonged time course compared with TSH, hence the name “Long Acting Thyroid Stimulator” (abbreviated LATS) 10. When the assay was adapted for use in mice, sera from some patients did not cross-react with the mouse thyroid and were LATS negative. The first commercialized TBI assay also involved antigen cross-reactivity, namely porcine TSHR 26. Correlation between TBI measured with the porcine TSHR versus the recombinant human TSHR is generally good (for example 27. In retrospect, the few exceptions, involving better recognition of porcine- than human-TSHR, likely represent antibody heteroclicity.
Parenthetically, we wish to comment on our present findings of relatively minor differences between TSAb measured in the human- and mouse-TSHR assays in the CXB recombinant inbred strains (BALB/c crossed with B6 mice). These data would appear to conflict with our previous report that, following human TSHR immunization, mouse-specific TSAb predominated over human-TSAb activity in BALB/c mice, and vice versa for B6 mice 16. The explanation for this apparent discrepancy is that the latter studies utilized BALB/c and B6 both of the Jackson substrain 16, whereas CXB mice are derived from the Bailey strains of BALB/c and B6. The clear-cut difference between BALB/c susceptibility versus B6 resistance to hyperthyroidism in Jackson-substrains 7,15 is muted in the Bailey-substrains 17. Moreover, in a direct comparison, fewer Bailey-BALB/c than Jackson-BALB/c (BALB/cByJ versus BALB/c/J) mice developed hyperthyroidism in response to TSHR A-subunit adenovirus immunization 28. The Bailey substrains of BALB/c and B6 mice have been bred separately from their Jackson counterparts for over 50 years 28
The most important outcome of our study, with potential implications for the genetics of human Graves’ disease, is the discovery of novel candidate genes besides the previously observed linkage to the MHC gene region 14,17. In BXH strains, preferential TSAb recognition of the mouse- versus human-TSHR in BXH mice was significantly linked to loci on Chr 12 between 110.245 and 111.769 Mb (LRS of 20.433; LOD of 4.43). This region includes the gene for type 3 thyroid deiodinase (Dio3), an enzyme involved in thyroid hormone inactivation 22. Increasing the power of genetic analysis by combining the TSAb species specificity data for the BXH and CXB mice increased the LOD score for this chr 12 region to >5.8.
Potentially of greater pathophysiological importance were loci slightly further upstream on Chr 12. Thus, LOD scores of 3.49 to 3.56 were obtained for the Chr 12 region from 113.270-120.329 Mb. Combining linkage data for the CXB and BXH strains narrowed this region to 113.595 -118.161 Mb. Remarkably, in this broad region, 32/66 (48%) of the genes markers encode the variable regions (IgV H) of immunoglobulin heavy chains (Supplementary Table S2), which are logical candidate genes for influencing antibody specificity. Moreover, the high frequency of VH genes in this locus, taken together with earlier findings that an IgV H gene controlled antibody heteroclicity (reactivity to steryl-oxazolone versus the immunogen furyloxazolone 29 ), strongly suggests that VH gene differences between mouse strains underlie the susceptibility (or lack thereof) to develop antibodies capable of activating the TSHR.
In early studies, it was reported that two genes, linked to immunoglobulin heavy chains (Gm) and HLA, controlled susceptibility to Graves disease 30. Because the chromosomal region encompassing genes for immunoglobulin heavy chain variable- and constant- regions is large (~ 3 million bases in mice 31), these observations for humans are potentially similar to ours in mice. However, similar associations were made for Hashimoto’s thyroiditis 32, a condition in which TSAb are absent. Moreover, later studies found no evidence for genes associated with the immunoglobulin H chain loci in Graves’ or Hashimoto’s diseases 33,34.
Why have immunoglobulin VH germline gene loci not previously been identified in genetic linkage and genome wide association studies in human Graves’ disease? In our opinion, ascertainment of traits for inclusion in these studies has been too broad. Greater stratification than a simple diagnosis of Graves’ disease is necessary. In Graves’ patients, TSHR antibodies are present at very low concentrations and are rarely (if ever) detected in family members without clinical disease. In addition, TSHR antibody levels are modulated by, and may disappear after, therapy with anti-thyroid drugs, surgery or radioiodine (for example 35,36). For these reasons, to our knowledge, no studies in humans have tested the genetic susceptibility for TSHR antibodies separately from hyperthyroidism, as has been done for autoantibodies to thyroid peroxidase and thyroglobulin 37. One advantage of the Graves’ animal model, using genetically identical mouse strains subject to identical environmental factors, is the ability to separate susceptibility to TSHR antibody development from variations in thyroid function 14,17. Our present data suggest that insight into the genetics of human Graves’ disease would be facilitated by restricting genetic analysis to untreated patients and which are subdivided according to TSAb levels or even divided into TSAb antibody subsets including perhaps differential TSAb activity for the human- versus the mouse-TSHR.
Besides our novel findings pointing to VH germline genes in Chr 12 as candidates for genetic susceptibility to TSAb generation, our present and previous linkage data 14,17 provide evidence for other loci underlying this trait:-
- TSHR antibodies (determined in TSAb and TBI assays) in CXB mice were linked to the same Chr 17 loci including MHC class I and II, complement components, tumor necrosis factor and lymphotoxin. These data support observations in humans in which MHC class II has long been associated with autoimmune thyroid disease (reviewed in 38) and very recently MHC class I 39.
- The highest LRS or LOD scores for TSAb species specificity were associated with the Chr 12 region including the gene encoding Deiodinase 3 (Dio3) 22, a seleno-enzyme that catalyzes the conversion of thyroid hormones to inactive metabolites. However, because it is difficult to envisage how Dio3 could contribute to the pathogenesis of Graves’ disease, TSAb activity is more likely linked to another closely associated, presently unidentified gene.
- TSAb generation in CXB mice linked to Chr X loci between 72.5 and 83.3 Mb, supporting our previous finding with TSHR antibodies detected in the TBI assay 17. There are no obvious candidate genes in this region, with the possible exception of interleukin 1 receptor accessory protein-like 1 (Il1rapl1). In addition to the foregoing distal Chr X loci, TSHR antibody linkage to more proximal Chr X loci (32-44 Mb) was noted for TBI in BXH mice 14. Again, there are no logical candidate genes in this region. As noted previously, although not confirmed in all data sets 40,41, associations with the X chromosome have been reported for human Graves’ disease 42-44.
In conclusion, measuring mouse-TSAb versus human-TSAb in mice with different and known genetic backgrounds provides novel insight into the genetic basis for susceptibility to induction of antibodies capable of activating the TSHR. Most important, we provide the first evidence suggesting involvement of immunoglobulin VH genes in this process. These data suggest that re-examination of these candidate genes in human Graves’ disease is warranted, using improved subject ascertainment and stratification, for example by including only patients with untreated disease of recent onset, of the same gender, and focusing on TSHR antibodies rather than a simple past or present history of disease.
FN2Conflict of Interest: The authors declare that no conflict of interest exists.
This work was supported by the National Institutes of Health Grants DK54684 (S.M.M.), DK19289 (B.R), U01AA13499, U24AA13513, P20-DA-21131, U01CA105417and U24RR021760 (R.W.W.). R.W.W. and GeneNetwork are also supported by Integrative Neuroscience Initiative on Alcoholism-National Institute on Alcohol Abuse and Alcoholism (INIA-NIAAA); Biomedical Informatics Research Network-National Center for Research Resources (BIRN-NCRR); Mouse Models of Human Cancers Consortium-National Cancer Institute (MMHCC-NCI) and a Human Brain Project. We are also grateful for contributions by Dr. Boris Catz, Los Angeles.
|IgV H||immunoglobulin heavy chain variable region|
|LRS||Linkage ratio statistic|
|RI mice||recombinant inbred mice|
|TSAb||thyroid stimulating antibody|
|TBI||TSH binding inhibition|
|TSHR||thyroid stimulating hormone receptor|
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|15.||Chen CR,et al. Susceptibility rather than resistance to hyperthyroidism is dominant in a thyrotropin receptor adenovirus-induced animal model of Graves’ disease as revealed by BALB/c-C57BL/6 hybrid miceEndocrinolYear: 200414549274933|
|16.||Misharin A,et al. Vitamin D deficiency modulates Graves’ hyperthyroidism induced in BALB/c mice by thyrotropin receptor immunizationEndocrinolYear: 200915010511060|
|17.||Aliesky HA,et al. Probing the genetic basis for thyrotropin receptor antibodies and hyperthyroidism in immunized CXB recombinant inbred miceEndocrinolYear: 200614727892800|
|18.||Chen CR,Aliesky HA,Guo J,Rapoport B,McLachlan SM. Blockade of costimulation between T cells and antigen-presenting cells: an approach to suppress murine Graves’ disease induced using thyrotropin receptor-expressing adenovirusThyroidYear: 20061642743416756463|
|19.||Williams RW,Gu L,Qi S,Lu L. The genetic structure of recombinant inbred mice: high-resolution consensus maps for complex trait analysisGenome BiolYear: 20012 Research0046.1-0046.18.|
|20.||Churchill GA,Doerge RW. Empirical threshold values for quantitative trait mappingGeneticsYear: 19941389639717851788|
|21.||Williams RW,Strom RC,Goldowitz D. Natural variation in neuron number in mice is linked to a major quantitative trait locus on Chr 11J NeurosciYear: 1998181381469412494|
|22.||Hernandez A,Martinez ME,Croteau W,St Germain DL. Complex organization and structure of sense and antisense transcripts expressed from the DIO3 gene imprinted locusGenomicsYear: 20048341342414962667|
|23.||Nagayama Y,McLachlan SM,Rapoport B,Niwa M. A major role for non-MHC genes, but not for micro-organisms, in a novel model of Graves’ hyperthyroidismThyroidYear: 20031323323812729471|
|24.||Mirza IH,Wilkin TJ. Antibody specificity in the immune response to insulinInt Arch Allergy Appl ImmunolYear: 1989892612632668202|
|25.||Minnerath JM,Wakem LP,Comfort LL,Sherman F,Jemmerson R. The BALB/c mouse B-cell response to pigeon cytochrome c initiates as a heteroclitic response specific for the self antigen mouse cytochrome cProc Natl Acad Sci U S AYear: 19959212379123838618905|
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|27.||Kakinuma A,et al. Comparison of recombinant human thyrotropin receptors versus porcine thyrotropin receptors in the thyrotropin binding inhibition assay for thyrotropin receptor autoantibodiesThyroidYear: 1999984985510524562|
|28.||Seetharamaiah GS,Land KJ. Differential Susceptibility of BALB/c and BALB/cBy mice to Graves’ hyperthyroidismThyroidYear: 20061665165816889488|
|29.||Makela O,Matoso-Ferreira A,Kaartinen M. Fine specificity of the immune response to oxazolones III. Antibodies but not contact sensitivity specific for 2-furyloxazolone are controlled by an Igh-V gene in the mouseEur J ImmunolYear: 198313101710226689303|
|30.||Uno H,Sasasuk T,Tamai H,Matsumoto H. Two major genes, linked to HLA and Gm, control susceptibility to Graves’ diseaseNatureYear: 19812927687706894965|
|31.||Retter I,et al. Sequence and Characterization of the Ig Heavy Chain Constant and Partial Variable Region of the Mouse Strain 129S1J ImmunolYear: 20071792419242717675503|
|32.||Tamai H,et al. Immunogenetics of Hashimoto’s and Graves’ diseasesJ Clin Endocrinol MetabYear: 19856062663838076|
|33.||Weetman AP,et al. Immunogenetics of Graves’ ophthalmopathyClin EndocrinolYear: 198828619628|
|34.||Barbesino G,Tomer Y,Concepcion E,Davies TF,Greenberg DA. Linkage analysis of candidate genes in autoimmune thyroid disease. I. Selected immunoregulatory genesJ Clin Endocrinol MetabYear: 199883158015899589659|
|35.||McGregor AM,et al. Carbimazole and the autoimmune response in Graves’ diseaseN Engl J MedYear: 19803033023076247656|
|36.||Chiovato L,et al. Disappearance of humoral thyroid autoimmunity after complete removal of thyroid antigensAnn Intern MedYear: 200313934635112965943|
|37.||Tomer Y,Greenberg DA,Barbesino G,Concepcion E,Davies TF. CTLA-4 and not CD28 is a susceptibility gene for thyroid autoantibody productionJ Clin Endocrinol MetabYear: 2001861687169311297604|
|38.||Jacobson EM,Tomer Y. The Genetic Basis of Thyroid AutoimmunityThyroidYear: 20071794996117824829|
|39.||Simmonds MJ,et al. A novel and major association of HLA-C in Graves’ disease that eclipses the classical HLA-DRB1 effectHum Mol GenetYear: 2007162149215317597093|
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Keywords: Graves’ disease, immunoglobulin heavy chain variable region, thyroid stimulating antibody, thyroid stimulating hormone receptor.
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Wagner tops Mount St. Mary’s 74-65 in NEC actionWednesday, Feb 6, 2013
The Seahawks (13-9, 7-4), who edged the Mountaineers 52-50 on Jan. 24, took a 38-26 halftime lead this time. Mount St. Mary’s (9-13, 4-7) outscored Wagner 39-36 in the second half, but it wasn’t enough.
Williams, who just missed a double-double with nine rebounds, got help from Latif Rivers, who scored 17 points. Josh Thompson chipped in 10.
For the Mountaineers, Rashad Whack scored 19 points, 15 coming on his 5-of-12 shooting from 3-point range. Kelvin Parker came off the bench to contribute 15 points and Shivaughn Wiggins added 11. But it wasn’t enough, not with Wagner dominating inside.
The Seahawks outscored Mount St. Mary’s 42-16 in the paint. They hit 21 of 27 foul shots, the Mountaineers 10 of 14.
- Freshman Eric Eaves scores 26, SC State beats UMES 68-63
- No. 3 Villanova too much for No. 11 Providence in 72-60 win
- No. 19 Louisville rolls past Boston College 79-47
- No. 9 Virginia pulls away from Pitt 64-50
- Forbes leads No. 10 Spartans to 89-73 rout of Michigan
- Garino scores 27 as George Washington beats VCU, 72-69


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'Season 4 reloaded' that's what Activision and Infinity Ward are calling the new Call of Duty Modern Warfare mid-season 4 updates. With a massive size of almost 36 GB, the game has made a special place in the hearts of fans and their hard drives. With exciting new game modes, loot, and an operator, this update seems to address the demand of fans. Activision and Infinity Ward have done an excellent job of keeping the game fresh yet close to its roots.
The immensely loved Warzone is introducing an increase in players for Battle Royale Quads bumping the number of players from 150 to 200. So players, brace yourself for a lot of action at the Superstore! A new game mode Juggernaut Royale has also been added, which will drop one Juggernaut Suit on the map; players who go into gulag will lose the suit but don't worry, look up towards the sky as another one will soon drop.
Spotter Scope is yet another new addition to warzone loot items. The scope is a reusable item enabling players to scan the environment just like a sniper scope, and for all of you crafty players, don't sweat, as it will have no optical glint helping you keep a low profile.
The gulag will be introducing new weapons with semi-auto rifles and snipers coming into the fray, and for all of you daredevils, there will also be a load out with just fists and throwing knives.
Modern Warfare multiplayer players can rejoice as a new map is going to be introduced. The map is called Cheshire Park and has been known as The Garden in previous leaks. This will be ideal for players who love close-quarters combat and mid-range flights.
Team Defender, a new mode, has also been introduced in multiplayer. It is a game mode that has made its way in the previous Call of Duty Franchises. The mode is similar to capture the flag, but there will be only one flag present rather than two. The player who holds the flag will earn the whole squad a point per second, the squad with the most point wins. so keep your friends close, and the flag closer.
New Blueprints and Operators
Rytec Amr is a new addition to the sniper rifle category. The gun can be unlocked by completing a challenge. It can also be bought from the store via the 'Lost Souls' bundle. The store will also be featuring a new operator, 'Roze.' Several other bundles Rogue Operative, Golden Idol, and Rifle Juliet blueprint, will also be added to the store.
These are exciting times, as many other subtle changes are also expected in the update. There are several rumors of changes in the Warzone map; many have speculated that the Stadium will now be open for players. Several guns will also be balanced with the Grau, and Mp5 receiving nerfs, which will shift the Meta quite a bit. Hopefully, this update will also fix several lags and server issues that have plagued the game since release. Players will be able to experience all of this and more as the update goes live on 30th June at 11:00 am Pakistan and 29th June 11:00 pm Pacific time.
The author is a gaming enthusiast and loves writing about technology.
Image credit: Activision
ISLAMABAD – Pakistani rupee remained stable against the US dollar as the market is looking for direction amid Pakistan’s ongoing negotiations with the International Monetary Fund (IMF).
Amid the economic uncertainty, the local currency remained unchanged against the greenback, moving up 0.01 percent during the opening hours of trading. Before noon, PKR was hovering at 285.40 in the interbank.
In contrast to the inter-bank, the dollar continued its upward trajectory in the open market and is reportedly available at Rs315, as the gap between the official rate and open market rate is Rs30 per dollar.
As the money market is waiting for a positive development from the economic front, International Monetary Fund (IMF) Mission Chief to Pakistan said they continued engagement with Islamabad focusing on the restoration of foreign exchange proper market functioning, with special on the upcoming budget.
KARACHI - The price of a single tola of 24-karat gold in Pakistan is Rs 232,700 on Tuesday.The price of 10 grams of 24k gold was recorded at Rs199,510.
Likewise, 10 grams of 22k gold were being traded for Rs163,624 while a single tola of 22-karat gold was being sold at Rs 190,880.
Note: The gold rate in Pakistan is fluctuating according to the international market so the price is never been fixed. The below rates are provided by local gold markets and Sarafa Markets of different cities.
|Lahore||PKR 232,700||PKR 2,500|
|Karachi||PKR 232,700||PKR 2,500|
|Islamabad||PKR 232,700||PKR 2,500|
|Peshawar||PKR 232,700||PKR 2,500|
|Quetta||PKR 232,700||PKR 2,500|
|Sialkot||PKR 232,700||PKR 2,500|
|Attock||PKR 232,700||PKR 2,500|
|Gujranwala||PKR 232,700||PKR 2,500|
|Jehlum||PKR 232,700||PKR 2,500|
|Multan||PKR 232,700||PKR 2,500|
|Bahawalpur||PKR 232,700||PKR 2,500|
|Gujrat||PKR 232,700||PKR 2,500|
|Nawabshah||PKR 232,700||PKR 2,500|
|Chakwal||PKR 232,700||PKR 2,500|
|Hyderabad||PKR 232,700||PKR 2,500|
|Nowshehra||PKR 232,700||PKR 2,500|
|Sargodha||PKR 232,700||PKR 2,500|
|Faisalabad||PKR 232,700||PKR 2,500|
|Mirpur||PKR 232,700||PKR 2,500|
Copyright ©2023. Reproduction of this website's content without express written permission from 'Daily Pakistan' is strictly prohibited.

domain: news


Controlling the Questions
The Republican presidential candidates made demands recently concerning the format of debates and the questions asked. They want more say over how the forums are handled.
Shop Talk panelist Jasmine Crighton said some of the debates have been poorly run and some of the questions were dumb. But she thinks the candidates are off-base with their demands.
Crighton also said the GOP candidates’ complaints have not been limited to the debates. For example, Ben Carson has complained about the “liberal media” going after him. Crighton said Carson and the others should expect follow up questions to statements they’ve made about their campaign and about their past.
Panelist Rich Egger said it’s time to retire the tired complaint about the “liberal media.” He said Democrat Hillary Clinton has probably been scrutinized more than any candidate running this year.
Panelist Jonathan Ahl said Carson and Donald Trump are the two biggest instigators of change in the debate format. He noted they’re also the two who’ve never served in public office and who’ve never had to stand up to public scrutiny.
Ahl doesn’t like the way the Republican debates have been divided into an under-card and a main event. He said some of the candidates in the large field are getting marginalized. Ahl said the candidates should be split randomly into smaller groups with debates held over the course of several nights.


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The sweat gland for the underarm is located in the armpit. This is why sweating and body odor mostly affects these areas. An armpit itch is an irritating sensation that makes one continuously want to scratch. Scratching might temporarily relieve the itch but scratching can also cause infection or contamination. Depending on the symptoms, like dryness, bumps, redness, discoloration, or swelling it can be associated with armpits itch.
It’s embarrassing when you’re in public and suddenly your armpits itch. Because of the sensitivity of armpits, it’s difficult to not scratch the itch. An armpit rash can last for hours or worse, days. Knowing the causes would help prevent future discomfort as most rashes can be easily treated. Armpit itch symptoms can be a result of the environment. But in some other cases, it might be due to more severe causes that would require different treatments.
Causes Why Armpits Itch:
1. Heat Rash
This is one of the most common causes of why armpits itch. The location of the armpits makes them susceptible to heat. Excess heat causes rashes. Thus, causing your armpits to itch.
Shaving can also cause rashes on your armpits. Using dull blades or blades that you’ve used more than three times can lead to itchy armpits. Dry shaving can also cause itching in your armpits because the skin needs to remain moisturized at all times. Running a blade along your dry skin would cause irritations and lead to itching.
It is easier for bacteria to collect and spread when your armpit is wet. Once bacteria spread along with your armpits, it becomes susceptible to infections that cause itchy armpits. Candida and yeast infection occur frequently in moist areas of the skin. In hot weather, because of heat or poor hygiene, Candida infection can become worse.
Some substances that contain strong chemicals can react with your skin causing itchy armpits. Substances like laundry detergents, soaps, deodorants, or even some type of clothing material can cause your skin to itch.
This is a rash that is caused by a microscopic itch rash. It enters the top layer of the skin where it feeds and this causes the skin to itch and have severe irritations. This mite is spread by direct contact with another infected person or by contact with infected furniture and beddings. The symptoms include severe itching especially in the night, rashes, and sores because of too much scratching.
Psoriasis is a kind of skin infection that causes an overgrowth of skin cells, creating an itchy and painful rash. It is considered a disorder where the body attacks its own cells.
Can armpits itch and not have rashes?
Rashes are usually accompanied with rashes. But it is possible to have itchy armpits without rash or any noticeable skin changes. Cases like these are usually harmless because they are often caused by temporary issues but on some occasions, severe health issues can cause itching with a rash.
Here are some things associated with itchy skin without rashes
- Dry skin: When the skin starts to lose moisture, it can begin to itch. Dry skin gives off a flaky or scaly look and feel.
- Bug bites: Bug bites can leave a small mold with a hole in the middle. They can make the skin around the bite feel itchy and irritated. Bugs like mosquitoes, bed bugs, lice, or spiders can cause itches like this.
- Medication side effects: Some medications can have side effects that include itching. This itching can occur without a rash.
- Skin cancer: skin cancers usually don’t have significant symptoms but some people can notice itching on skin lesions as an early symptom.
- Lymphoma: This is cancer that affects the lymphatic system. It can cause swellings of the lymph nodes. Lymph nodes are found in the armpits, groin, and neck. A type of lymphoma is Hodgkin’s and non-Hodgkin’s lymphoma. A percentage of people that have this cancer are affected by itching. This itching is not accompanied by a skin rash.
- Cancer: Breast cancer develops on breast cells. A rare type of breast cancer is inflammatory breast cancer which is accompanied by your armpits itching.
How Do I Stop My Armpits To Itch?
Having itchy armpits can really give discomfort. Here are ways to stop your armpits from itching:
- Avoid using soaps with harsh chemicals, they can cause dry skin.
- Apply moisturizer on your skin regularly especially after a bath.
- Practice good personal hygiene. Keep your underarms clean especially after doing a physical activity that involves sweating.
- Wear loose clothing so that your armpits can breathe and sweat won’t pool on your armpits.
- Use talcum powder during hot weather to stop heat rashes from occurring.
- Don’t shave your armpits often, especially if you have sensitive armpits. Give your armpits time to recover from the previous shave.
- Use natural, fragrance-free detergents that don’t contain chemicals that could cause skin irritations.
Armpits are very sensitive parts of the body. It is important to determine the cause so as to rule out a serious condition and proffer an appropriate solution. Poor hygiene, allergic reactions, or dermatitis are the most causes of armpit rashes. Once you determine the cause of the problem, you must follow the treatment plan properly to reduce the risk of a more severe complication.
A more serious issue could be cancer. Symptoms that accompany cancer include swelling, redness, warmth, and other noticeable skin changes. It is advisable to consult your doctor if you notice any itch that seems more than just a normal itch.

domain: medical


4peaks is described as '4Peaks - For Peaks, Four Peaks. The award winning 4Peaks helps molecular biologists to visualize and edit their DNA sequence trace files. Analyzing your sequence has never been so much fun!' and is an app in the Education & Reference category. There are more than 10 alternatives to 4peaks for a variety of platforms, including Windows, Mac, Linux, Online / Web-based and Self-Hosted solutions. The best alternative is DNADynamo. It's not free, so if you're looking for a free alternative, you could try Chromas or Serial Cloner. Other great apps like 4peaks are Benchling (Free Personal), SnapGene Viewer (Free Personal), Geneious (Paid) and CLC Genomics Workbench (Paid).
GeneStudio Pro is a modern suite of molecular biology applications for the Windows platform built on our sequence format conversion engine, SeqVerter. All manipulations with sequences of different formats are thus seamless and transparent for the user.
QIAGEN CLC Main Workbench is ideal for DNA, RNA, & protein sequence data analysis with a wide variety of features, no advanced computer skills required! The workbench is available for Windows, Mac OS X, and Linux platforms.
BVTech Plasmid is DNA sequence analysis and plasmid drawing software for Windows PCs.You can use it to plan your DNA cloning, draw high quality plasmid maps, analyse your DNA sequencing data, align sequences, and much more.
Use SimVector to draw plasmid maps, perform restriction analysis and mapping. The plasmid drawing software also simulates cloning experiments such as gateway cloning, ta cloning and restriction cloning


Protest against 'illegal mining' enters fifth day
Residents of Chakkmirpur village on the Punjab-Himachal Pradesh border continued their protest against alleged illegal mining for the fifth day on Saturday.Updated: Jul 22, 2012 00:24 IST
Residents of Chakkmirpur village on the Punjab-Himachal Pradesh border continued their protest against alleged illegal mining for the fifth day on Saturday.
Protesters say that the mining takes place in Himachal Pradesh, but heavy vehicles carrying the output pass through their village, causing environmental pollution. They also alleged that roads in the area have not been built to carry such heavy vehicle load.
Village sarpanch Kamlesh Devi said: Vehicular traffic is a big threat to the environment of all villages in Punjab near the mining site in Himachal Pradesh. More than 100 vehicles run on roads that are unfit for carrying such loads."
She added: "Large-scale illegal mining has been going on in the neighbouring state. In gross violation of norms, digging more than 50-feet is rampant in the area, causing serious threat of floods."
Surender Singh, a village resident, said: "The mining has affected the life of residents in ten villages including Chakkmirpur, Tote, Sathwan, Hangwal, Ulaha, Tode Katwal, Bela, Saryala and Jalalpur."
He added: "If not stopped immediately, the situation could lead to a disaster in the area in the rainy season." Ravinder Singh, another resident, said: "The administration has not shown any signs of even listening to their demands. We will be forced to step up the agitation, if our demand of diverting the traffic and stopping illegal mining is not met,"
Block development and panchayat officer (BDPO), Yudhvir Singh said: "The main concern of residents is to stop the plying of trucks from their villages."He added that he had apprised SDM Rahul Chaba about the situation, and the administration would soon find a solution to the problem.
However, Chaba said that he was not aware of the matter. "I will talk to the BDPO concerned to understand the villagers' demands and will act accordingly."
First Published: Jul 22, 2012 00:20 IST


text:
AYER — Most students look forward to getting on the bus to go home when school lets out, but not the after-school robotics program participants at Ayer Middle School.
On Tuesday afternoons, under the supervision of Steve Branam, a dedicated group of students meets in the art room to explore the science of robotics. Branam, a parent of two children in the Ayer schools, is a software engineer with Juniper Networks and uses the Lego Robotics Invention System to introduce this topic to the interested students.
“The Lego kit is great because all kids have used Lego at one point or another, so it is a familiar medium for them,” Branam said.
The students start by building a creation from Legos. They may design something on their own or utilize one of several diagrams found in the Lego Robotics text. After the construction phase, students then add computer boards, motors and sensors to make their creations more interactive, and software to create programs that allow the robot to perform functions such as turning, raising an arm or making a noise.
“My goal is to introduce students to the basic concepts of programming in general, and then move into the specifics of programming their robot,” Branam said. “This is a great hands-on learning situation. They learn to plan how they are going to build something, build it, and then problem-solve using the experimental method. If parts are falling off the robot, then the students know they need to rework something. If their robot is not responding to commands as expected, then the students know they need to reprogram.”
Students involved in the program have built devices that can follow along a wall, using technology similar to that used in the popular Roomba device, a robotic vacuum cleaner, made by IRobot Corp., that sweeps the floor with no human assistance. All of the kids receive hands-on experience doing real-world technical work.
Afterschool efforts such as the robotics program let students try out different disciplines to see if they might be interested in pursuing one further.
“I do this program twice a year so I can allow more students to experience it,” Branam said, noting that students seem to like the instant feedback they receive.
“If their robot doesn’t work correctly, they know immediately that they need to redesign or adapt their device,” the engineer said.
And the students seem to enjoy their time with Branam. Student Karen Bouchard said she enjoys her experience working with Branam and that she looks forward to the Tuesday afternoon meetings of the robotics club.
“When I was a little kid, Santa would always get me Lego stuff and I always liked it,” Bouchard said. “I heard about this robotics thing and how it uses Legos, so I wanted to try it.”

domain: design


text:
If you have a HSA, you cannot have a medical FSA, but you can open a limited purpose flexible spending account (L-FSA) to use for dental and vision expenses. Dependent care and transportation and parking FSAs are still allowed. Consider a L-FSA if you contribute the annual maximum to your HSA. This will allow you to save your HSA dollars for future healthcare expenses, but also set aside pre-tax dollars in your L-FSA for current year vision and dental expenses.
Have more questions? Submit a request


A bold staffer at University of South Africa has staged a nude protest after being forced to resign for declining sexual advances from her boss.
In Kenya, most if not all women occupying powerful office have gained career progression from sleeping with their bosses. From the likes of Caroline Mutoko who used to sleep with her boss Patrick Quarcoo before being edged out by Grace Msalame, to the likes of Sheila Mwanyigha whose lack of brains still got her a job, women have shown us alot.
It is the reason productivity in Nairobi’s workforce has declined because sexual considerations has taken precedence over merit.
These same women who happily handed themselves over to their bosses for jobs, then mutate into motivational quote-hawkers after being consumed by guilt in their later years. They join toxic feminist movements portraying all men as predators, yet it’s the few who occupy powerful office who are rotten in their heads.
Today we have seen a nominee in Mike Sonko’s cabinet turn down her appointment because she was being compelled to sleep with him in return. That is the kind of news we want to hear. Women of moral and standing saying no to bullies. If Sonko wants pussy, there’s many places he can get it beginning with our Instagram fashion bloggers and socialites who are out there hawking ass to the highest bidder.
Beautiful women have used their pussies to gain a head-start over their much lesser endowed female counterparts, who tend to be more meritorious. Indeed, the not so hot chics in school tended to be the smarter ones.
But because of using pussy, beauty, shameless self-promotion and the likes, some women have gone on to “succeed” in this grossly manipulated work environment, but later on come to blame this imbalance to “men” yet they themselves aided in this vice.
The men too of this country have lost direction as they have thrown caution to the wind and allowed their penises to dictate corporate or even Government policy. Many of the new age “tenderprenuers” are women whose only claim to fame is sleeping with the various heads of parastatals, companies and the likes.
To see women like this one take a bold move to decline sexual advances is an inspiration which all girls in Kenya should emulate. Instead of following the toxicity of latter day-saints like Caroline Mutoko who’ve benefited from such malpractice in-order to gain a head-start in life and concede their morals for short term benefit, they should emulate this South African lady.
These women who have benefited from using their pussy for career progression and then later on turn to hawkers of motivational quotes and bible verses, are enemies who young girls shouldn’t emulate at all. They should be given a wide berth.
Would you like to get published on this Popular Blog? You can now email Cyprian Nyakundi any breaking news, Exposes, story ideas, human interest articles or interesting videos on: [email protected] Videos and pictures can be sent to +254 710 280 973 on WhatsApp, Signal and Telegram.


text:
Parts of the shaft date back to the 15C, but the present Cross is largely a 1885 reconstruction by Sydney Mitchell, based on drawings and engravings of the original. The earlier Mercat Cross was removed in 1756 as a traffic obstacle: its site is marked in the pavement, just a few metres from its present location. The Cross is embellished with the Scottish Royal Coat of Arms and the arms of Edinburgh, the Burgh of the Canongate, Leith, England, Scotland, Wales and Ireland, all in true heraldic tinctures. Public proclamations are made from the gallery three days after proclamation in London (that being the time a horseman takes to travel between the two capitals).
Architect: Sydney Mitchell
Building Date: 15C -1885

domain: history


text:
Andrew Lloyd Webber Initiative to Support Theatre Education Grant Award
New Hanover County Schools is pleased to announce that the American Theatre Wing, best known as the creator of the Antoinette Perry “Tony” Awards, is awarding the Snipes Academy of Arts and Design with a $16,000 Classroom Resource Grant from the Andrew Lloyd Webber Initiative. Snipes was one of nearly 200 applicants and one of 14 award winners of the prestigious grant received across 40 states.
Snipes will use the $16,000 Classroom Resource Grant to purchase equipment for their theater program. Support from the Initiative will allow for expansion of the theater program to produce work in a fully equipped space with working microphones, light, sound and expanded set design. The students will be able to perform quality shows with costumes while building their skills in theatrical design and performance.
“This recognition from the American Theatre Wing is a tremendous honor for our school,” said Rachel Manning, principal of Snipes Academy of Arts and Design. “We are extremely excited about the opportunities this will afford our students, as it will provide them with all the necessary materials to completely produce high-quality theatrical productions.”
With the Andrew Lloyd Webber Foundation’s help, the American Theatre Wing is investing in public schools to help drama teachers get more of the resources they need to provide quality drama instruction in their schools. Through this Classroom Resource grant-giving program, funding is provided directly to under-resourced public schools to provide instruments, dance floors, lighting grids, and other necessary materials to help create new and enhance existing theater programs.
“Arts education, and making theater available to all young persons, is vital to the growth of our culture and society. The grant proposals received this year have shown the widespread need for arts funding in primary and secondary schools across America. I am proud that we are now entering the second year of this program, and am excited for the years to come,” said Andrew Lloyd Webber.
For more information about the Snipes Academy of Arts and Design theatre program contact Principal Rachel Manning at 910-251-6175, Rachel.Manning@nhcs.net or visit their website at http://www.nhcs.net/snipes/.
The American Theatre Wing, Heather Hitchens, President and CEO) is dedicated to advancing artistic excellence and nurturing theatre’s next generation: on the stage, behind the scenes, and in the audience. For a century, the Wing has pursued this mission with programs that span the nation to invest in the growth and evolution of American Theatre Traditionally, the Wing has encouraged members of the theatre community to share their off-stage time and talent directly with the theatre audience at large–whether it was singing for the troops in the Stage Door Canteen of the 1940s, or sharing their stories on a podcast today. As the founders of The Tony Awards®, the American Theatre Wing has developed the foremost national platform for the recognition of theatrical achievement on Broadway. Yet the Wing’s reach extends beyond Broadway and beyond New York. The Wing develops the next generation of theatre professionals through the SpringboardNYC and Theatre Intern Network programs, incubates innovative theatre across the country through the National Theatre Company Grants, fosters the song of American theatre through the Jonathan Larson Grants, honors the best in New York theatrical design with the Henry Hewes Design Award, and illuminates the creative process through the “Working in the Theatre” program and media archive.
More information about the ALW Classroom Resource Grants can be found at
http://americantheatrewing.org/program/the-andrew-lloyd-webber-initiative/. For more information on the Andrew Lloyd Webber Initiative and the American Theatre Wing, visit AmericanTheatreWing.org and Facebook.com/TheAmericanTheatreWing and follow @TheWing on Instagram and Twitter.
Photos from the 2017 Snipes Holiday Performance

domain: artistic


text: We leveraged IDH wild type glioblastomas (GBM) and derivative neurosphere models to define the tumor-intrinsic mRNA transcription phenotype. We found that intratumoral heterogeneity is reflected in the transcriptome and associated with increased tumor microenvironment presence. We performed in silico cell sorting to demonstrate that M2 macrophages/microglia are the most frequent type of immune cells in the glioma microenvironment, followed by CD4+ T lymphocytes. Longitudinal transcriptome analysis of 124 pairs of primary and recurrent glioma pairs showed expression subtype is retained in 53% of cases with no proneural to mesenchymal transition being apparent. Inference of the tumor microenvironment through gene signatures revealed a decrease in invading monocytes but a subtype dependent increase in M2 macrophages/microglia cells after disease recurrence. Our longitudinal expression dataset can be accessed at http://ackbar.cnio.es:3838/RecuR/. Our study provides a comprehensive transcriptional and cellular landscape of IDH wild type GBM during treatment modulated tumor evolution.


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At the urging of Ryan Adams, whose adventures in cooking through all of Fergus Henderson’s Nose to Tail: The Whole Beast are chronicled at Nose to Tail at Home, I submitted a proposal for a weekly feature at the now-defunct Eat Me Daily (EMD) blog. My idea was to show how easy it is to make ingredients you would normally buy at a supermarket. I had already written here about bacon and pickles, but my sample article was about making mayonnaise, inspired by my first attempt while constructing my homemade BLT.
My proposal was never evaluated; the editorial staff at EMD all moved on to more prestigious gigs (missed it by that much). So here, for your consideration, is the first article in my failed series:
I used to work in the convenience food industry, so I know how simple it is to make many of the products found on supermarket shelves. Take away the packaging, and what youâ€™re left with is something you can make at home for less money and just a little time. Iâ€™ll show you how to make superior versions of supermarket standbys in your own kitchen.
Mayonnaise: More Than Just Sandwich Glue
Why bother to make mayonnaise? Because itâ€™s more than just a condiment.
Most of us all grew up with Hellmanâ€™s (Best Foods west of the Rockies), and probably have a jar in the fridge. Itâ€™s a perfectly good product, worth having around for quick sandwich-making, but a bit on the bland side. If you really want to â€œbring out the bestâ€ for salads or sauces, then you need to make your own.
It couldnâ€™t be easier; you only need five ingredients: an egg yolk, a cup of oil (preferably a neutral vegetable oil like canola or safflower), two teaspoons of lemon juice, a teaspoon of water, and half a teaspoon of salt. Everything should be at room temperature.
Whisk the yolk, water, lemon juice, and salt together in a large bowl.
While whisking the yolk mixture, add in a few drops of oil at a time until the emulsion forms. Keep whisking, and add the rest of the oil in a thin stream. Place something under the bowl (a dishtowel or potholder) to keep it from sliding as you whisk. The first time you try this you should whisk it by hand so you can get a feel for the process, but an immersion blender with a whisk attachment is a perfectly acceptable substitute. (You can still claim that you whisked it by hand, after all, a whisk and your hand were involved.)
In a few minutes you should have thick, creamy mayonnaise (and a very sore arm). Whisk in more water if you prefer a thinner texture.
If your mayonnaise never comes together or turns into a runny mess in the bowl, donâ€™t panic â€” itâ€™s merely broken, not ruined. To recover, add a teaspoon of water to a clean bowl and gradually start whisking in the broken mayo bit by bit until it re-emulsifies.
What To Do With It
If you treat your homemade mayo as an ingredient rather than a finished product, you can turn it into any number of sauces and garnishes by adding a few extra ingredients. If you start with olive oil and add a teaspoon of minced garlic to the yolk and liquid, you wind up with aioli. Add more lemon juice and some dill and you have a perfect sauce for salmon. Substitute tarragon for the dill and you have a base for chicken salad. Add more lemon juice and some minced shallot and you have a sauce for vegetables. Use lime juice instead of lemon and add minced chile pepper to make a killer sandwich spread. The possibilities are limited only by your imagination.
Why bother to make mayonnaise? Because itâ€™s more than convenient, itâ€™s essential.
Just donâ€™t get me started on Miracle Whip.

domain: food


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About pm kisan samman nidhi problem
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Earlier, the application for Pradhan Mantri Kisan Samman Nidhi Yojana/pm Kisan was done with the help of an offline nodal agency or by Lekhpal but currently, a new option was added on the official website of Pradhan Mantri Kisan Samman Nidhi Scheme pmkisan.gov.in Due to which farmers can get themselves registered or through Vasudha Center,. PM Kisan Samman Nidhi Scheme 2022 (PM-KISAN) is fully funded by the central government and is expected to cost around Rs. PM Kisan Samman Nidhi 2022 Rejected List..
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“I couldn’t have asked for a better way to have spent the past eight years”: MG to exit Grill Team
Mark Geyer has confirmed that he’ll be sleeping in for the next few years as he joins Triple M’s NRL coverage full time from 2018.
MG has been the tallest member of the Triple M Grill Team since it began in 2009 and following its 2000th show celebrations last week he’s decided it’s the right time to shift his focus to his two greatest passions in life – Family and Footy!
“My daughter Rafferty was a baby when I started breakfast radio, she’s now 10 and I’ve never been able to drop her at school,” MG told The Grill Team.
“I couldn’t have asked for a better way to have spent the past eight years; with our Triple M listeners and The Grill Team family! I’ll certainly miss the daily laughs but I’m really excited for a new adventure with Triple M NRL,” he said.
MG’s represented his country and is well known for being one of NSW’s Origin hard men. He proudly boasts a Grand Final win the Penny Panthers and has agreed to head up Triple M’s NRL coverage long term.
Happily, though, he’ll still be appearing regularly with the Grill Team throughout the footy season.
MG will call footy each weekend on Triple M’s national NRL coverage as well as star in a new NRL based weekday show.
The Triple M bosses couldn’t be happier with one station representative saying, “We couldn’t be happier!”


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Hello! Thank you for telling me that, I’m glad you like it :D I added the drawing on my shop for you, here. You can buy it with free shipping and $5 Off Each Item thru April 13. I hope you will get what you want. Thanks!
THANK YOU VERY MUCH!
Here, thank you for telling me that! :D


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Last Updated on June 19, 2019, by eNotes Editorial. Word Count: 409
Manilo Argueta's novel One Day of Life is hard to read. It doesn't have big words or complicated themes or a convoluted plot; it's a difficult text because it tells the story of life under a repressive, violent dictatorship. In this way, it's similar to Aleksandr Solzhenitsyn's story One Day...
(The entire section contains 409 words.)
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Manilo Argueta's novel One Day of Life is hard to read. It doesn't have big words or complicated themes or a convoluted plot; it's a difficult text because it tells the story of life under a repressive, violent dictatorship. In this way, it's similar to Aleksandr Solzhenitsyn's story One Day in the Life of Ivan Denisovich. In that one, though, the violence is mostly psychological. Here, in Argueta's novel, the violence is in your face, relentless and overpowering. We won't summarize the book's contents here. Rather, we'll analyze it. You should read it, and you should check out the excellent study guide available on this website.
In 1979, junior officers of the Salvadoran army deposed the violent and repressive regime of President Carols Romero. That coup d'etat set off a civil war, in which opposing sides were backed by the US and the USSR. That made El Salvador a proxy battleground in the Cold War, which is why the Salvadorans involved became irreconcilable. Left-wing and communist insurgents backed by the Soviet Union fought right-wing paramilitary groups allied with the army and backed by America. Death squads, child soldiers, and atrocities were commonplace.
Into this whirlwind came One Day of Life. It was published in 1980, the first year of the civil war, so it wasn't about the war. It was about living with the physical and emotional trauma inflicted on ordinary people by the dictatorships and broken politics of the post-Second-World-War period. It was certainly apropos of the time, and it has since become associated with the civil war, but it was more about that war's antecedents.
The story's central irony is emblematic of El Salvador's plight. Lupe and Jose work on a wealthy landowner's plantation, and they become involved in the leftist resistance, which destroys their livelihood and their family. You can't tell oppressed people not to fight, but when fighting destroys what they were trying to save, you have to ask why. That's the big question Argueta's novel doesn't really address. In this, it's typical of other stories to come out of Central America in that period, such as I, Rigoberta Menchu. They depict the suffering of peasants and well-meaning opponents of dictatorial governments, but they don't resolve their central themes or conflicts. That might just be the most important observation of all, because the countries of Central America turned out exactly this way. The civil wars are over, but the oppression and despair and violence and resistance continue.

domain: library


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The proposed closure has been designed to coincide with the completion of Arla’s new Arla’s new 1bn litre ‘super dairy’ in Aylesbury, Buckinghamshire.
In anticipation of the closure, which has been scheduled for April 2014, processing duties will be gradually moved from the Ashby-de-la-Zouch plant to the new Aylesbury facility. The phased transfer of raw milk from the Ashby to Aylesbury will begin in October 2013.
“Our new dairy in Aylesbury is on track to be processing milk, commercially, by the autumn and volumes are ahead of our original expectations,” said head of Arla Foods UK’s supply chain, Lars Dalsgaard.
“In line with our long-term business goal, we intend to maximise the use of our well invested dairies, complemented by our new, cutting edge, processing facility in Aylesbury, to meet the requirements of our customers.”

domain: economics


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Whether or not you and your accomplice are large gift-givers, the vacations are at all times a pleasant time to point out one another a bit additional love (if for no different motive than the vacations are additionally a super-busy, aggravating season). Letting her or him know you care doesn’t must require large bucks or tons of planning, both—you need horny, enjoyable, considerate, and quick, proper?
Here are a couple of simple, last-minute, scorching concepts to get you began.
Date Night Stocking
Choose an exercise you’ll be able to share in collectively, like dinner and a film, plus dessert afterward. (There are present playing cards only for all that!) Make it particular by placing the present playing cards in numbered envelopes with candy messages inside.
Everyone wants some self-care, so give your S.O. sufficient spa love for each of you (as a result of self-care is doubly good while you do it collectively). Start with a shower bomb to drop within the tub so you’ll be able to each soak and chill out, add some face masks, physique scrub, and oil for afterward. Scrub them down and oil them up. Your night in will be so enjoyable… it will also be very sensual.
Party in a Stocking
Craft cocktails are a factor proper now, so get a pocket bartending information and pair it with components for a superb time. Learn to make Moscow Mules collectively—you’ll be able to even embody the cups. Bonus factors if you happen to get them customized.
Day of Adventure
This one could take some work, however it may be so rewarding for you each. Choose a couple of actions relying on the place you reside and your pursuits. Do you’ve gotten mountains to hike? A forest for picnicking? A seashore to stroll? Plan out an in depth day and write about it in a card. Choose small objects that improve or decorate your journey: sun shades, a cute water bottle, an engraved compass, a map—get inventive! Your journey ought to final till sunset… and possibly longer.
This one actually will get the purpose throughout. Head into your favourite grownup retailer for treats to take pleasure in along with your lover underneath the tree, menorah, or mistletoe. Things like lube, a small vibe, cock rings, therapeutic massage oil, assorted condoms, a e-book of erotica, a DVD to observe collectively, perhaps even a blindfold. (Just make certain they’ve privateness to open this one, or it might be the household vacation horror story for years to come back.)
Remember, these are all simply beginning factors, and also you’ll want to make use of what you recognize about your accomplice’s likes and dislikes to make them really feel particular. Gift playing cards are nice, quick, and handy, however be sure to’re not simply handing them over in an envelope. It’s been confirmed time and time once more it’s the thought that counts, so put some thought into it, and make this a really particular vacation for the lover in your life.

domain: hobby


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Clematis need regular pruning to encourage flowers to bloom and keep them to a manageable size. Look at the the pruning groups for advice.
How to feed a Sundaville. If you grow Sundaville in containers, feed regularly during the growing season to stimulate new flowers.
Slugs and snails can cause a lot of damage to garden plants, especially to new growth. To minimize slug damage, why not grow plants that slugs don’t like to eat.
Pruning Erysimum ‘Bowles Mauve’ (wallflower). The plant flowers almost all year-round. So may be tricky to prune it.
Propagating mint is an easy way to get new mint plants. How to propagate mint from cuttings. Follow the instructions step-by-step.
Japanese anemones are valuable plants. They bring wonderful, soft colours to the garden. There are many good varieties.

domain: agronomy


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Cass is a beautiful black and orange lady who is oh-so-sweet! Friendly and curious, this lovely lady is also quite a purr machine. She is a big fan of ear scratches and pets and would love to find herself in a loving home with a cozy bed to nap in! Since Cass is a Golden Girl, she qualifies for our Sponsored by Simba program's Seniors-for-Seniors offer: no adoption fee for any pet 8years and up by an adopter 65 years plus. We hope you will come meet Cass as soon as you can.
*This program also covers Pets for Veterans: no adoption fee for any military veteran adopting a pet 1 year and older.
You’ve got questions and we’ve got you covered.
Pepper is a lovely Golden Girl. This little lady is so nice. She loves to sit high up so she can sit and watch what's happening down below. Pepper seems to do well with other cats in the room. Be…Learn more
Athena is a beautiful and petite brown tabby. She is a new arrival, and we are still getting to know this young girl. Please come meet Athena as soon as you can.Athena qualifies for our Sponsored by …Learn more
Olive is a beautiful black cat with stunning yellow eyes. She is wanting her future family to come by and whisk her away to a loving home. Hurry over to meet this 1-year-old kitty today and you'l…Learn more
All rights reserved. © 2023 Brandywine Valley SPCA.

domain: pet


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According to Ruth O'Regan, MD, BCI is the first genomic assay to demonstrate benefit from ovarian suppression.
In an analysis of the SOFT trial (NCT00066690), the Breast Cancer Index (BCI) test accurately identified premenopausal women undergoing adjuvant endocrine therapy for early-stage, hormone receptor (HR)–positive breast cancer who would derive benefit from adding ovarian function suppression (OFS).1
Ruth O’Regan, MD, chair of medicine and Charles A. Dewey Professor at the University of Rochester, presented the findings during the 2022 San Antonio Breast Cancer Symposium (SABCS). “This is the first genomic assay to demonstrate benefit from ovarian suppression, supporting additional clinical utility and value and validation of BCI and premenopausal women,” she said.
SOFT, a landmark international, three-arm, non-blinded, randomized clinical trial, included 3066 premenopausal women with HR-positive breast cancer. Patients were randomized 1:1:1 to receive tamoxifen alone, tamoxifen plus OFS, or exemestane plus OFS, all for 5 years. Data from that trial showed a 3% improvement in 12-year freedom from distant recurrence (DRFI) with exemestane plus OFS compared with tamoxifen alone (HR, 0.75; 95% CI, 0.59-0.97).
However, OFS can be toxic, and patients can struggle with compliance. Therefore, it is crucial to identify which patients are most likely to benefit, O’Regan said. Data from SOFT showed that women younger than 35 years appear to derive benefit from OFS following chemotherapy. Furthermore, investigators have developed a continuous composite that incorporates risk value age, nodal status, tumor size, grade, estrogen receptor (ER) status, progesterone receptor status, and Ki-67 that can assist with estimating the individual risk-based benefit of escalating endocrine therapy for HER2-negative patients.
“However, today, no genomic biomarker has been available to predict the benefit of errands of ovarian suppression premenopausal patients,” O’Regan said. “This is critical because of not just the short-term toxicities, but also long-term toxicities, which may not already be fully established.”
To develop such a biomarker, O’Regan and her colleagues evaluated the predictive and prognostic ability of the BCI test in patient tumor samples (n = 1717) collected from the SOFT trial.
The BCI is a genomic assay that incorporates molecular grade index (MGI) and the HOXB13/IL17BR (H/I) index ratio. In ER-positive breast cancer, the BCIN+ further incorporates tumor size and grade for N1 disease. BCI is prognostic for patents with N0 disease who received endocrine therapy with or without chemotherapy and BCIN+ is predictive for those with N1 disease.
“BCI has been found to be prognostic in determining individualized risk of late recurrences and also cumulative overall distant recurrence from year 0 to year 10,” O’Regan said.
The BCI predictive part is focused on the (H/I) ratio and is exclusively on the ability of the (H/I) AI to assess question signaling pathways. “What it does is report the likelihood of benefit from extended endocrine therapy as a ‘Yes,’ if the BCI (H/I) is high or ‘No’ if the BCI (H/I) is low,” O’Regan said.
The primary study objective was determining whether BCI (H/I) status was predictive for benefit with exemestane plus OFS vs tamoxifen. Investigators hypothesized that BCI (H/I)-high status would be predictive for OFS benefit, while BCI (H/I) low status would not.
In the data presented at SABCS, investigators assessed tumor samples collected from 573 patients assigned to tamoxifen, 551 assigned to tamoxifen plus OFS, 563 assigned to exemestane. The median follow-up was 12 years.
In the BCI analysis cohort (n = 1687), half (50.1%) of patients had grade 2 tumors, 25.4% had grade 1, 22.9% had grade 3, and tumor grade was unknown in 1.5%. Patient age ranged from younger than 35 years (11.3%) to older than 50 years (10.6%). More than 65% of patients had pN0 nodal status, 25.3% had pN+ 1 to 3 status, and 9.0% had pN+ 4+ status. Tumor size was equal to or less than 2 cm in 64.1% of patients, greater than 2 cm in 34.4%, and was unknown in 1.5%.
For HER2 status, 11.7% of patients were positive. Most patients (53.3%) received chemotherapy. O’Regan said the BCI analysis cohort was representative of the parent SOFT trial.
Investigators determined that 58% of cancers were BCI (H/I) low and 42% were BCI (H/I) high. There was a significant treatment by biomarker interaction for exemestane plus OFS vs tamoxifen (P <.01). The interaction was not as strong for tamoxifen plus OFS vs tamoxifen (P = .16).
In the overall HR-positive cohort, those with BCI (H/I)-high tumors, patients did not derive an absolute benefit for exemestane plus OFS vs tamoxifen (HR, 1.03; 95%, 0.70-1.53) or tamoxifen plus OFS vs tamoxifen (HR, 1.05; 95% CI, 0.72-1.54). The absolute benefit was –0.4% for exemestane plus OFS and –1.2% for tamoxifen plus OFS.
In contrast, investigators observed an absolute benefit of 11.6% for exemestane plus OFS vs tamoxifen (HR, 0.48; 95%, 0.33-0.71) in the BCI (H/I)-low group. For tamoxifen plus OFS vs tamoxifen, the absolute benefit was 7.3% (HR, 0.69; 95% CI 0.48-0.97).
In patients with HR-positive, HER2-negative tumors, BCI (H/I) was again predictive in the low group. The absolute 12-year benefit was 13.2% for exemestane plus OFS vs tamoxifen (HR, 0.39; 95% CI, 0.25-0.60) and 7.4% for tamoxifen plus OFS vs tamoxifen (HR, 0.64; 95% CI, 0.44-0.93).
In the BCI (H/I) group, the absolute benefit was –0.2% for exemestane plus OFS vs tamoxifen (HR, 1.03; 95% CI, 0.64-1.68) and –6.8% for tamoxifen plus OFS vs tamoxifen (HR, 1.37; 95% CI, 0.89-2.13).
Investigators also found that predictive performance was generally consistent across subgroups, including nodal status, prior chemotherapy, and age. Furthermore, consistent with data reported from the SOFT trial, the benefit derived from tamoxifen plus OFS was smaller than the benefit derived from exemestane plus OFS.
“BCI risk scores were prognostic in premenopausal women with HR-positive tumors receiving adjuvant endocrine therapy [with or without] chemotherapy in that higher BCI risk scores were associated with the worse outcome,” O’Regan said. “The BCI (H/I) was predictive of ovarian suppression benefit. However, contrary to our study hypothesis, the BCI (H/I)-low group consistently derived clinically meaningful benefit while the BCI (H/I)-high group did not. We’re not sure what these results mean, but it does point to potential differences in tumor biology underlying the ovarian suppression response.”
O’Regan R, Zhang Y, Fleming GF, et al. Evaluation of the Breast Cancer Index in premenopausal women with early-stage HR+ breast cancer in the SOFT trial. Presented at: 2022 San Antonio Breast Cancer Symposium; December 6-10, 2022; San Antonio, TX. Abstract GS1-06.


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Backed by our troop of talented workforce, we are engrossed in manufacturing, exporting and supplying premium array of Laboratory/Lab Glass Beakers. Under the strict vigilance of our experienced professionals, offered beakers are manufactured with utmost perfection by utilizing premium quality glass and sophisticated techniques. These beakers are highly acclaimed in professional labs and can be heated directly in an open flame. Used for mixing and gentle heating, the provided Laboratory Beakers can easily be accessed by our prestigious customers at highly discounted prices.
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Mobile : 918045478663

domain: chemistry


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I don't believe that being Celiac or having a food intolerance or allergy is the end of ever enjoying a good meal or a scrumptious dessert again. My focus is on creating bakery and products that taste awesome, have a nice texture, a decent shelf life and have nutritious value.
Everything I create is with a purpose and good intention not just for my family, but for you and your family also.
In 2014, my life changed dramatically when I found out gluten was the culprit playing havoc on my health. My holistic doctor had suggested I give it up for two weeks to see how I felt. It turned out that within three days I was a totally different person. I'd never felt so good. Not willing to eat it for two months to be tested we decided to just say I am gluten intolerant though I do have classic symptoms of Celiac. Shortly after, my husband, Rick decided to become gluten free and found he felt better.
There aren't any bakeries that offer anything gluten free and what we found in stores was dry, made with preservatives, using mega white rice flour and tapioca starch. And so Bell Farmstead Bakery & Products was born.
It was important to my family that not only are we eating something that tastes good, but it had to have some degree of nutritional value. So I learned to create gluten free flour blends instead of using packaged gluten free blends from the store. Incorporating ancient grains and grains that our ancestors would have grown and used; amaranth, sorghum, millet and oat. I started experimenting with flaxseed meal, coconut, almond and garbanzo bean flours. All of these flours/meal brought depth, texture, unique flavor and nutrients to our baked goods. Using these blends I spend a lot of time creating the recipes that we are all enjoying. My family and I found using organic and non-gmo ingredients not only made everything we ate healthier but enhanced the flavor!
Our chickens are started on a feed special ordered from Virginia that is organic and no soy. They graduate to a layer feed that is no soy, no wheat. The extras they get is organic produce which some is grown by me. The chickens supply the eggs for the bakery.
Bell Farmstead Bakery & Products ingredients are organic, if not organic; non-gmo. No soy. No corn. No peanuts. No xanthan gum.
Bell Farmstead Bakery & Products will do its very best to work with other food intolerance and allergies.


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Democratic gubernatorial candidate Beto O’ Rourke has always known his campaign was going to be an uphill battle. Despite his multiple efforts to maintain a neutral warzone, ‘cancel culture’ is still out to get him.
In his first statewide campaign, when he challenged Ted Cruz for a U.S. Senate seat, O’Rourke charmed his way through Texas, visiting towns other politicians of both parties wouldn’t waste time in.
Always with the vision to empower Democrats in small red towns to create lasting change in their communities. Sadly, O’Rourke lost his race by an unexpectedly small margin.
O’Rourke’s second time around, has been even more difficult.
His gubernatorial campaign is still hitting the road, talking to conservatives in rural places in Texas where no other Democrat would visit. However, unlike his first statewide campaign, where there were no protests or disturbances, O’Rourke is literally getting run out of town by Republicans.
According to Texas Monthly, this past weekend, in a small community an hour north of San Antonio, the O’Rourke campaign, hoping to hold a town hall, tried and failed to secure the use of four different event venues.
When news got out that O’Rourke was coming to visit, Canyon Lake residents of Comal County went into a frenzy, boycotting his campaign before he even had placed a foot in town.
O’Rourke was first set to speak at Maven’s Inn & Grill, but after locals threatened to boycott the event, his campaign was forced to change places. So it set its eyes on Canyon Lake High School.
However, officials with the Comal Independent School District said the district did not, as a rule, allow rallies to take place on school grounds. So O’Rourke looked for a third venue, Canyon Lake Resource and Recreation Center, who also backtracked because of “safety issues.”
The last place his campaign tried was Whitewater Amphitheater, and after that offer was also rescinded, his campaign gave up plans for Canyon Lake and instead held the rally at New Braunfels.
During the event, O’Rourke said he dreamed of a government that was “not so interested in the culture wars” and more interested in making Texans’ lives better.
But, his little incident in Comal County proves that he can’t escape them.
O’Rourke talked about reaching out to Texans of all political persuasions, to a crowd that was almost solely made up of his loyal Democratic supporters – since he was unable to deliver his message to the more diverse audience of Canyon Lake.
In the end, according to Texas Monthly, O’Rourke urged his fellow party members to assert their presence and to say forcefully that “we live here and we have a different point of view than other people do.”
Showing up will wear down the stigma of being Democrats, which will “give people permission to believe.”


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The Miraculous Journey of Edward Tulane
Saturday, September 30, 2023 | 11:00 AM
Thomasville Center for the Arts
A vain and haughty china rabbit battles the high seas, befriends a happy hobo and comforts a sick child as he discovers the transformative power of love. Based on Kate DiCamillo's award-winning 2006 novel for children. Appropriate for all ages. FREE admission.

domain: entertainment


Police statistics not impressive
THE EDITOR, Sir:
I am quite sure no one was fooled by the attempt of acting Deputy Commissioner of Police Glenmore Hinds to get us to believe that the Jamaica Constabulary Force achieved major operational successes in 2009.
The acting deputy commissioner needed to be more specific in the way he presented the figures. For example, Hinds said the police apprehended 993 wanted persons in the period. The gentleman needed to clarify what he meant by 'apprehended'. Did he mean 'arrested and charged', or 'arrested and released without being charged'? This clarification becomes more urgent in light of the fact that he later stated that 32,923 persons were arrested and charged in the same period.
WANTED FOR WHAT?
The ongoing 70 per cent of unsolved murders in this country sends chills up the spines of all of its citizens. It would, therefore, be comforting if the acting deputy commissioner had told us how many of the 32,923 and the 993 persons arrested and charged/ apprehended were murder cases.
We notice that he is basking in what he calls "increased police presence". But those of us who still live in Jamaica are unable to verify such a claim since we don't see the police after certain hours. Besides, has this "increased police presence" translated into the arresting and charging of even 30 per cent of those who murdered over 1,600 of our brothers and sisters in 2009? Not to mention the tens of thousands who were murdered over the past many years?
Instead of coming up with meaningless figures, the acting deputy commissioner and his colleagues need to start crafting and effecting crime-fighting measures to eliminate serious crimes in Jamaica, especially murder. How much longer must the nation wait on these men to get it right?
I am, etc.,