text:
Barrett’s Esophagus is considered to be a precursor to adenocarcinoma and the information on VDR expression in normal and Barrett’s esophagus is significantly lacking. In this study, we examined the expression of VDR in the lower esophagus and gastric cardia of normal and Barrett’s esophagus by immunofluorescence. Columnar mucosa but not squamous mucosa at the gastroesophageal junction showed positive immunofluorescence to VDR. Submucosal glands and ducts deep to the normal squamous mucosa stained positive for VDR and localized in the cytoplasm and perinuclear regions with no nuclear staining. Interestingly, the Barrett’s mucosa stained strongly positive for VDR. Glandular structures in the mucosal layer were far less abundant in the Barrett’s mucosa than in the normal gastric mucosa. As a result, fewer structures deep to the Barrett’s epithelial layer stained positive for VDR when compared to normal gastric mucosa. These findings suggest that in normal esophagus VDR expression is restricted to columnar epithelium and glandular structures. Furthermore, strong VDR expression in Barrett’s mucosa may indicate an increased sensitivity of this tissue to endogenous or therapeutic effects of Vitamin D.
Barrett’s esophagus; Epithelium; Gastroesophageal junction; Vitamin D; Vitamin D receptor
A potent immunomodulatory role of Vitamin D in both innate and adaptive immunity has recently been appreciated. In allergic asthma, activation of NF-κB induces transcription of various cytokines and chemokines involved in allergic airway inflammation. The nuclear import of activated NF-κB p50/RelA subunit is dependent on importin α3 (KPNA4) and importin α4 (KPNA3). In this study, we examined the role of importin α3 in immunomodulatory effect of calcitriol in human bronchial smooth muscle cells (HBSMCs).
Cultured HBSMCs were stimulated with calcitriol in the presence and absence of cytokines, TNF-α, IL-1β, and IL-10. The mRNA transcripts of importin α3 and α4 were analyzed using qPCR while protein expression of importin α3, α4 and nuclear RelA was analyzed by immunoblotting.
Calcitriol significantly decreased mRNA and protein expression of importin α3 as well as nuclear protein expression of NF-κB p65 (RelA). The decreased activation of RelA by calcitriol was confirmed by decreased release of RelA-inducible molecules, including IL-5, IL-6 and IL-8, by HBSMCs upon calcitriol treatment. Calcitriol attenuated the effect of TNF-α and IL-1β to upregulate mRNA and protein expression of importin α3. IL-10 significantly decreased the TNF-α induced expression of importin α3 and this effect was further potentiated by calcitriol.
These data suggest that under inflammatory conditions, calcitriol decreases the expression of importin α3 resulting in decreased nuclear import of activated RelA. This could be a novel mechanism by which calcitriol could exert its immunomodulatory effects to reduce allergic airway inflammation and thus may alleviate the symptoms in allergic asthma.
Active vitamin D; Allergic airway inflammation; Asthma; Bronchial smooth muscle cells; Calcitriol; Importinα3 (KPNA4); Importinα4 (KPNA3); NF-κB; Vitamin D receptor (VDR)
Esophageal adenocarcinoma carries a poor prognosis. Tumor response to neoadjuvant therapy is a key prognostic factor in patients with adenocarcinoma of the esophagus, but is inconsistent. Identifying tumor characteristics that portend a favorable response to neoadjuvant therapy would be a valuable clinical tool. The anticancer actions of vitamin D and its receptor may have implications. In this study, 15 biopsy specimens were procured retrospectively from patients being treated for adenocarcinoma of the esophagus. The tissue was immunostained for the vitamin D receptor and compared on the basis of response to neoadjuvant therapy. Tumors that did not respond to neoadjuvant therapy had greater expression of VDR than tumors that responded completely. Expression of VDR declined with tumor de-differentiation. The data suggest that a relationship between vitamin D receptor expression and response to neoadjuvant therapy is plausible.
Adenocarcinoma; Esophageal cancer; Neoadjuvant therapy; Vitamin D; Vitamin D receptor
Asthma is one of the most common chronic inflammatory diseases of the airways. Calcitriol exerts its action through Vitamin D receptor (VDR), which is a high affinity nuclear receptor. VDR is a transcription factor that alters the transcription of target genes which are involved in a wide spectrum of biological responses. Lower serum vitamin D levels are associated with airway hyperresponsiveness and increased asthma severity. Prohibitin is a ubiquitously expressed protein localized to the cell and mitochondrial membranes and the nucleus.
Methods and results
HBSMCs were cultured and treated with calcitriol and/or TNF-α. The mRNA and protein expression of prohibitin and VDR were analyzed using qPCR and immunoblotting, respectively. In the in vivo studies, female BALB/c mice were fed with special vitamin D-deficient or 2,000 IU/kg of vitamin D-supplemented diet for 13 weeks. Mouse model of allergic airway inflammation was developed by OVA-sensitization and challenge. The expression pattern of TNF-α, prohibitin and VDR in the lung of OVA-sensitized mice was analyzed using immunofluorescence. Calcitriol significantly increased and TNF-α decreased the protein and mRNA expression of prohibitin and VDR in HBSMCs. There was significantly increased expression of TNF-α and decreased expression of VDR and prohibitin in the lung of vitamin D-deficient mouse model of allergic airway inflammation.
These results suggest that under inflammatory conditions there is decreased expression of VDR resulting in decreased expression of prohibitin, which is a vitamin D target gene. Vitamin D deficiency causes increase in the expression of TNF-α, thereby increasing inflammation and decreases the expression of VDR and prohibitin. Supplementation with vitamin D reduces the levels of TNF-α, thereby increasing the expression of VDR and prohibitin that could be responsible for reducing allergic airway inflammation.
Allergic airway inflammation; Asthma; Human Bronchial Smooth Cells; Inflammation; Prohibitin; TNF-α; Vitamin D receptor; Vitamin D
Vitamin D is a sectosteroid that functions through Vitamin D receptor (VDR), a transcription factor, which controls the transcription of many targets genes. Vitamin D deficiency has been linked with cardiovascular diseases, including heart failure and coronary artery disease. Suppressor of cytokine signaling (SOCS)3 regulates different biological processes such as inflammation and cellular differentiation and is an endogenous negative regulator of cardiac hypertrophy.
The purpose of this study was to test the hypothesis that vitamin D deficiency causes cardiomyocyte hypertrophy and increased proinflammatory profile in epicardial adipose tissue(EAT), and this correlates with decreased expression of SOCS3 in cardiomyocytes and EAT.
Eight female Yucatan miniswine were fed vitamin D-sufficient (900 IU/d) or vitamin D-deficient hypercholesterolemic diet. Lipid profile, metabolic panel, and serum 25(OH)D levels were regularly measured. After 12 months animals were euthanized and histological, immunohistochemical and qPCR studies were performed on myocardium and epicardial fat.
Histological studies showed cardiac hypertrophy, as judged by cardiac myocyte cross sectional area, in vitamin D-deficient group. Immunohistochemical and qPCR analyses showed significantly decreased mRNA and protein expression of VDR and SOCS3 in cardiomyocytes of vitamin D-deficient animals. EAT from vitamin D-deficient group had significantly higher expression of TNF-α, IL-6, MCP-1, and decreased adiponectin in association with increased inflammatory cellular infiltrate. Interestingly, EAT from vitamin D-deficient group had significantly decreased expression of SOCS3.
These data suggest that vitamin D deficiency induces hypertrophy in cardiomyocytes which is associated with decreased expression of VDR and SOCS3. Vitamin D deficiency is also associated with increased inflammatory markers in EAT. Activity of VDR in the body is controlled through regulation of vitamin D metabolites. Therefore, restoration of VDR function by supplementation of VDR ligands in vitamin D- deficient population might be helpful in reducing inflammation and cardiovascular risk.
Cardiac Hypertrophy; Coronary Artery Disease; Epicardial Fat; Inflammation; Vitamin D
Gene therapy has attracted attention for its potential to treat several cardiovascular diseases. The use of adeno-associated viral (AAV) vectors to facilitate therapeutic gene transfer to suppress intimal hyperplasia is a promising concept. The objective of this study was to analyze the in vivo transduction of a novel recombinant AAV-2/9 vector with SM22α promoter, containing β-galactosidase gene (Lac Z) or green fluorescent protein (GFP) as reporter genes, to the medial layer smooth muscle cells (SMCs) of swine coronary and peripheral arteries.
The AAV2/9 vector containing SM22α (1×1013 pfu) were administered into carotid/femoral/coronary arteries of domestic swine using irrigating balloon catheter-based gene delivery. Following gene transfer, cryosections of arteries were processed for X-Gal and GFP analysis. Fluorescence microscopy and Western blotting were done to analyze the GFP expression in the SMCs.
LacZ mRNA expression was visualized in the medial layer 7 days after vector administration. The GFP expression was detected at 7th day and lasted for at least 2 months showing the longer-lasting expression of the AAV2/9-vector. Control arteries did not show any expression of GFP or LacZ. There was no significant effect of AAV2/9 viral transduction on serum amylase, fibrinogen and serum CRP levels.
These finding support the use of AAV2/9 as a vector to effectively transduce a gene in SMCs of coronary and peripheral arteries without causing inflammation.
Adeno-associated virus; Gene therapy; Intimal hyperplasia; Restenosis; Vascular smooth muscle cells; Vector
Migration to draining lymph nodes is a critical requirement for dendritic cells (DCs) to control T-cell–mediated immunity. The calcium-activated potassium channel KCa3.1 has been shown to be involved in regulating cell migration in multiple cell types. In this study, KCa3.1 expression and its functional role in lung DC migration were examined. Fluorescence-labeled antigen was intranasally delivered into mouse lungs to label lung Ag-carrying DCs. Lung CD11chighCD11blow and CD11clowCD11bhigh DCs from PBS-treated and ovalbumin (OVA)-sensitized mice were sorted using MACS and FACS. Indo-1 and DiBAC4(3) were used to measure intracellular Ca2+ and membrane potential, respectively. The mRNA expression of KCa3.1 was examined using real-time PCR. Expression of KCa3.1 protein and CCR7 was measured using flow cytometry. Migration of two lung DC subsets to lymphatic chemokines was examined using TransWell in the absence or presence of the KCa3.1 blocker TRAM-34. OVA sensitization up-regulated mRNA and protein expression of KCa3.1 in lung DCs, with a greater response by the CD11chighCD11blow than CD11clowCD11bhigh DCs. Although KCa3.1 expression in Ag-carrying DCs was higher than that in non–Ag-carrying DCs in OVA-sensitized mice, the difference was not as prominent. However, Ag-carrying lung DCs expressed significantly higher CCR7 than non–Ag-carrying DCs. CCL19, CCL21, and KCa3.1 activator 1-EBIO induced an increase in intracellular calcium in both DC subsets. In addition, 1-EBIO–induced calcium increase was suppressed by TRAM-34. In vitro blockade of KCa3.1 with TRAM-34 impaired CCL19/CCL21-induced transmigration. In conclusion, KCa3.1 expression in lung DCs is up-regulated by OVA sensitization in both lung DC subsets, and KCa3.1 is involved in lung DC migration to lymphatic chemokines.
allergic airway inflammation; antigen uptake; asthma; calcium-activated potassium channel; dendritic cell
Leukemia/lymphoma related factor/POK erythroid myeloid ontogenic factor (LRF/Pokemon) is a member of the POK family of proteins that promotes oncogenesis in several forms of cancer. Recently, we found higher LRF expression in human breast and prostate carcinomas compared to the corresponding normal tissues. The aim of this study was to examine the regulation of LRF expression in human prostate cells. Epidermal growth factor (EGF) and its receptors mediate several tumorigenic cascades that regulate cell differentiation, proliferation, migration and survival of prostate cancer cells. There was significantly higher level of LRF expression in the nucleus of LNCaP and PC-3 cells than RWPE-1 cells. A significant increase in LRF expression was observed with increasing doses of EGF in more aggressive and androgen-sensitive prostate cancer cells suggesting that EGF signaling pathway is critical in upregulating the expression of LRF/Pokemon to promote oncogenesis.
prostate cancer; Pokemon; EGF; LRF; ZBTB7
Vascular restenosis following coronary artery bypass graft can cause major clinical complications due to intimal hyperplasia in venous conduits. However, the precise underlying mechanisms of intimal hyperplasia are still unclear. We have recently reported that increased expression of connexin43 (Cx43) is involved in the proliferation of vascular smooth muscle cells (SMCs) in human saphenous vein (SV). In this study, we investigated the signaling transduction pathway involved in Cx43 expression and SV SMC proliferation. Angiotensin-II (AT-II, 100ng/ml) increased angiotensin II receptor 1 (AT-1R) protein expression and insulin-like growth factor-1 (IGF-1) (100ng/ml) upregulated IGF-1 receptor (IGF-1R) protein expression in SV SMCs. Interestingly, AT-1R expression was also increased by IGF-1 treatment, and IGF-1R expression was increased by AT-II treatment, which was blocked by siRNA-IGF-1R and siRNA-AT-1R respectively. Furthermore, the effect of AT-II and IGF-1 signal cross-talk inducing upregulation of their reciprocal receptors was blocked by siRNA against extracellular signal–regulated kinases 1/2 (Erk 1/2) in SMCs of saphenous vein. Moreover, AT-II and IGF-1 induced Cx43 expression via phosphorylation of Erk 1/2 and activation of transcription factor activator protein 1 (AP-1) through their reciprocal receptors in SV SMCs. These data demonstrate a cross-talk between IGF-1R and AT-1R in AT-II and IGF-1-induced Cx43 expression in SV SMCs involving Erk 1/2 and downstream activation of the AP-1 transcription factor.
Angiotensin II; Connexins; Intimal Hyperplasia; Restenosis; Vascular Smooth Muscle Cells; Vein Graft Disease
The growth and differentiation of cells is regulated by cytokines by binding to cell-surface receptors and activating intracellular signal transduction cascade. Suppressor of cytokine signaling (SOCS)-3 is a negative regulator of cytokines. In this study we examined the expression of SOCS3 in porcine coronary artery smooth muscle cells (PCASMCs) in vitro and in proliferating smooth muscle cells of neointimal lesions after coronary artery intervention in a swine model.
Methods and Results
PCASMCs were cultured and stimulated with TNF-α and/or IGF-1 individually or in combination. Protein expression of SOCS-3 was examined using Western blot. For in vivo studies, six female Yucatan miniswine were fed with special high cholesterol diet for 8 months. At 4 months of high cholesterol diet, animals underwent coronary balloon angioplasty. At the end of 8 months animals were euthanized, coronary arteries were isolated and morphological and histological studies were performed. Western blot data revealed significantly high SOCS-3 expression in PCASMCs in the presence of either TNF-α or IGF-1 (5–6 fold) alone. However, in the presence of both TNF-α and IGF-1 the SOCS-3 expression was significantly decreased (4–5 fold). Results from morphological studies including, H&E and Masson’s trichrome stain showed typical lesions with significant neointimal proliferation. Histological evaluation showed expression of smooth muscle α-actin and significantly increased proliferating cell nuclear antigen (PCNA) in neointimal lesion. Interestingly, there was significantly decreased expression of SOCS3 in smooth muscle cells of neointima as compared to control.
These data suggest that SOCS3 expression is decreased in proliferating smooth muscle cells of neointimal lesions. This leads to uncontrolled growth of vascular smooth muscle cells in injured arteries leading to restenosis. Therefore, local delivery of SOCS3 gene at the site of injury after coronary artery intervention could regulate the proliferation of vascular smooth muscle cells and help in preventing the neointimal hyperplasia and restenosis.
Angioplasty; Coronary Intervention; Inflammation; Intimal Hyperplasia; Restenosis; Suppressor of cytokine signaling-3
Cytokines released by the immune cells at the site of plaque milieu induce smooth muscle cell apoptosis to promote plaque instability. But, neuropeptide Y (NPY), a pleotropic factor, may modulate the effects of cytokines in atherosclerotic plaques of patients with carotid stenosis. Our aim was to investigate the relative expression of NPY-Y1, NPY-Y2 and NPY-Y5 receptors on carotid plaque vascular smooth muscle cells (pVSMCs) of symptomatic (S) and asymptomatic (AS) patients and examine the effect of inflammatory cytokines on the expression of NPY receptors, that may attenuate plaque rupture.
Methods and Results
In healthy carotid artery, there was significantly increased immunopositivity and increased mRNA transcripts of NPY-Y1 and NPY-Y5 receptors in thin sections and isolated VSMCs, respectively, compared to S and AS plaques. However, the NPY-Y2 expression was higher in S and AS pVSMCs than controls. Stimulation of the cells with TNF-α, IL-12 or IFN-γ (50 ng/ml) decreased mRNA transcripts of NPY-Y1 and NPY-Y5 and increased NPY-Y2 mRNAs in VSMCs of healthy carotid artery. The effect of the cytokines on mRNA transcripts of NPY-Y5 and NPY-Y2 in pVSMCs of S and AS patients was similar to healthy VSMCs, but with variable effect on NPY-Y1.
Increased expression of NPY-Y2 receptors in symptomatic pVSMCs than in healthy and asymptomatic subjects suggests a potential role of NPY-Y2 in plaque instability. This is further supported by the pronounced effect of atheroma-associated cytokines to increase NPY-Y2 mRNA transcripts in pVSMCs of patients with carotid stenosis.
Atheromatous cytokines; Atherosclerosis; Carotid plaque; Carotid stenosis; Neuropeptide Y; Smooth muscle cells
Angiopoietin (Ang)1 and Ang2 are ligands for Tie2 tyrosine kinase receptor (Tie2). Elevated levels of Ang1 and Ang2 in induced sputum of patients with asthma have been reported, with a positive correlation of Ang2 levels with the severity of airway occlusion. Although studies have shown Tie2-mediated regulation of nonvascular cells in some pathological conditions, current knowledge on Tie2 signaling in asthma is limited to the vasculature. We examined the expression pattern of Ang1, Ang2, vascular endothelial growth factor (VEGF), and Tie2 and their correlation with the degree of airway remodeling in the lung of ovalbumin (OVA)-sensitized and OVA-challenged mice with airway hyperresponsiveness. Lung tissues were isolated from Balb/c mice after OVA sensitization and challenge. Hematoxylin and eosin, periodic acid-Schiff, and trichrome staining were used to show the lung pathology. The expression of Ang1, Ang2, VEGF, and Tie2 was examined using immunofluorescence, Western blot, ELISA, and real-time PCR. In the lung of normal mice, Tie2 expression was detected only in the blood vessels. However, in the lung of OVA-sensitized and OVA-challenged mice, Tie2 was abundantly expressed in airway epithelial cells and in a subset of macrophages in addition to constitutive expression in pulmonary vessels. The increase in Tie2 expression correlated with the severity of airway remodeling. Macrophages and airway epithelial cells express Ang2 and VEGF only in allergic models. Ang1 was constitutively expressed, with a decrease in mRNA level in allergic models. In conclusion, increased expression of Tie2 and Ang2 in allergic airway epithelium and alveolar macrophages correlates with the severity of airway remodeling.
airway epithelial cells; airway remodeling; angiopoietins; Tie2 tyrosine kinase receptor; vascular endothelial growth factor
Apoptosis of vascular smooth muscle cells (SMCs) is controlled by a balance between the effect of growth factors and cytokines, and is involved in plaque instability in advanced atherosclerotic lesions. Recently, we reported high levels of atheroma-associated cytokines, including tumor necrosis factor-α (TNF-α), in carotid plaques of symptomatic patients. These cytokines induce apoptosis of vascular SMCs, and thus could be responsible for plaque rupture, a clinically devastating event. In this study, we examined the effect of TNF-α on the cell cycle inhibitor p27kip and apoptosis of SMCs in human carotid plaques, and the underlying mechanism. Both Forkhead box subclass o1 (FoxO1) and p27kip were more strongly expressed in symptomatic than asymptomatic atherosclerotic plaques. TNF-α significantly induced the expression of FoxO1 in asymptomatic plaque SMCs in a dose- and time-dependent manner via JNK signaling pathway. TNF-α also induced phosphorylation of FoxO1, resulting in its cytoplasmic translocation/nuclear exclusion of transcription factors. The effect of TNF-α was blocked by the PI3K inhibitor, LY294002. Meanwhile, TNF-α not only induced the p27kip expression and cell cycle arrest in G0–G1 phase, but also enhanced caspase-3 activity and induced apoptosis in SMCs of asymptomatic plaques. However, the potential effect of TNF-α on the cell cycle inhibitor p27kip and apoptosis of SMCs was inhibited by siRNA against FoxO1 in asymptomatic patients. These data suggest the involvement of FoxO1 transcription factor in TNF-α-induced expression of a cell cycle regulatory protein and apoptosis of SMCs, thus regulating the stability of atherosclerotic plaques with carotid stenosis.
Apoptosis; Atherosclerosis; Forkhead Transcription Factor; Plaque Stability; Vascular Smooth Muscle Cell
We previously reported that Fms-like tyrosine kinase 3 ligand (Flt3-L) reversed airway hyperresponsiveness (AHR) and airway inflammation, and increased the number of regulatory CD11chighCD8αhighCD11blow dendritic cells and CD4+CD25+ICOS+Foxp3+IL-10+ T-regulatory cells in the lung of allergen-sensitized and -challenged mice. In this study, we evaluated the effect of Flt3-L on Th17 cells and expression of suppressors of cytokine signaling (SOCS) proteins in the lungs of house dust mite (HDM)–sensitized and –challenged mice. BALB/c mice were sensitized and challenged with HDM, and AHR to methacholine was established. Mice were treated with Flt3-L (5 μg, intraperitoneal) daily for 10 days. Levels of IL-4, -5, -6, -8, and -13, and transforming growth factor (TGF)–β in the bronchoalveolar lavage fluid (BALF) were examined by ELISA. Flt3-L treatment reversed existing AHR to methacholine and substantially decreased eosinophils, neutrophils, IL-5, -6, -8, and IL-13, and TGF-β levels in the BALF. HDM-sensitized and -challenged mice showed a significant increase in lung CD4+IL-17+IL-23R+CD25− T cells with high expression of retinoic acid–related orphan receptor (ROR)–γt transcripts. However, administration of Flt3-L substantially decreased the number of lung CD4+IL-17+IL-23R+CD25− T cells, with significantly decreased expression of ROR-γt mRNA in these cells. HDM sensitization caused a significant increase in the expression of SOCS-1, -3, and -5 in the lung. Flt3-L treatment abolished the increase in SOCS-1 and SOCS-3 proteins, whereas SOCS-5 expression was significantly reduced. These data suggest that the therapeutic effect of Flt3-L in reversing the hallmarks of allergic asthma in a mouse model is mediated by decreasing IL-6 and TGF-β levels in the BALF, which, in turn, decrease CD4+IL-17+IL-23R+ROR-γt+CD25− T cells and the expression of SOCS-1 and SOCS-3 in the lung of HDM-sensitized and -challenged mice.
airway hyperresponsiveness; house dust mite; retinoic acid–related orphan receptor–γt; suppressors of cytokine signaling; T helper cell type 17
We recently reported that the adoptive transfer of T-regulatory cells (Tregs) isolated from lung and spleen tissue of green fluorescent protein–transgenic mice reversed airway hyperresponsiveness and airway inflammation. Because Programmed Death-1 (PD-1) is a pivotal receptor regulating effector T-cell activation by Tregs, we evaluated whether PD-1 is involved in the therapeutic effect of naturally occurring Tregs (NTregs) and inducible Tregs (iTregs) in cockroach (CRA)-sensitized and challenged mice. The CD4+CD25+ NTregs and CD4+CD25− iTregs isolated from the lungs and spleens of BALB/c mice were adoptively transferred into CRA-sensitized and CRA-challenged mice with and without anti–PD-1 antibody (100 μg/mice). The CD4+CD25+ T cells in the lung were phenotyped after adoptive transfer. Concentrations of IL-4, IL-5, IL-10, IFN-γ, and IL-13 in bronchoalveolar lavage fluid (BALF) were measured using ELISA. The NTregs and iTregs from either lung or spleen tissue reversed airway hyperresponsiveness for at least 4 wk. However, the therapeutic effect was blocked by administering the anti–PD-1 antibody. The administration of Tregs-recipient mice with anti–PD-1 antibody significantly decreased cytotoxic T-lymphocyte antigen-4 expression, with low concentrations of Forkhead-winged transcriptional factor box 3 (Foxp3) mRNA transcripts in lung CD4+CD25+ T cells. These mice had substantially higher concentrations of BALF IL-4, IL-5, and IL-13, but significantly decreased levels of BALF IL-10. Adoptive therapy recipients without the anti–PD-1 antibody exhibited high levels of CTLA-4 expression and Foxp3 transcripts in lung CD4+CD25+ T cells, with a significant decrease in BALF IL-4, IL-5, and IL-13 concentrations and a substantial increase in BALF IL-10 concentrations. These data suggest that the reversal of airway hyperresponsiveness and airway inflammation by Tregs is mediated in part by PD-1, because other costimulatory molecules (e.g., inducible costimulatory molecule [ICOS] or CTLA-4) have been shown to play a role in Treg-mediated suppression.
airway hyperresponsiveness; airway inflammation; anti–PD-1 antibody; cockroach antigen; Forkhead-winged transcriptional factor box P3
Coronary revascularization by coronary artery bypass grafting (CABG) is recommended in patients with recurrent myocardial ischemia. However, the long-term results of CABG using saphenous vein (SV) graft, compared to internal mammary artery (IMA) graft, have not been satisfactory. The SV graft failure is due to development of intimal hyperplasia, a process characterized by abnormal migration and proliferation of smooth muscle cells (SMCs) in the intimal layer of the vein graft. Insulin growth factor 1 (IGF-1) is a major mitogenic growth factor released at the site of the shear stress-induced graft injury. This study, for the first time, compares the extent of IGF-1-PI3K-Akt activation in isolated human by pass graft conduits. Human SV and IMA vessels were collected and SMCs isolated and cultured. In cultured SMCs, effect of IGF-1 was examined on total and phosphorylated PI3K, Akt and IGF-1R by Western blot analysis. Cell proliferation was measured using BrdU ELISA. There was no significant difference in the basal expression of phosphorylated PI3K, Akt and IGF-1R in SV and IMA SMCs from human bypass conduits. However, we observed an upregulation of IGF-1 receptors in the SV SMCs in response to IGF-1 stimulation with no effect in IMA SMCs. Furthermore, the immunoblotting and cellular activation of signaling ELISA (CASE) assay demonstrated a significantly higher activity of both PI3K and Akt in IGF-1-stimulated SV SMCs than IMA. This was inhibited by an IGF-1R blocking antibody. IGF-1 induced proliferation in both SV and IMA SMCs was inhibited by a PI3K inhibitor, wortmannin. These data demonstrate differential activity of IGF-1-induced PI3K-Akt activation, which was quantitatively and temporally greater in SV SMCs than in the IMA. This, at least in part, could explain the greater propensity of the SV conduits than the IMA to undergo intimal hyperplasia following CABG.
Coronary artery bypass graft; IGF-1; Internal mammary artery; Intimal hyperplasia; PI3K; Akt/PKB; Proliferation; Restenosis; Saphenous vein; Vascular smooth muscle cells
We previously reported in an ovalbumin-induced model of allergic asthma that Fms-like tyrosine kinase 3 ligand (Flt3-L) reversed airway hyperresponsiveness (AHR) and airway inflammation, and increased the number of regulatory CD11chighCD8αhighCD11blow dendritic cells in the lung. In this study, we investigated the effect of Flt3-L in a clinically relevant aeroallergen-induced asthma on the phenotypic expression of lung T cells. Balb/c mice were sensitized and challenged with cockroach antigen (CRA), and AHR to methacholine was established. These mice received three intraperitoneal injections of anti-CD25 antibody (PC61; 250 μg) and Flt3-L (3 μg) daily for 10 days. Cytokines and Ig levels in the serum were measured and differential bronchoalveolar lavage fluid (BALF) cell counts were examined. Flt3-L reversed AHR to methacholine to the control level. Flt3-L significantly decreased levels of BALF IL-5, IFN-γ, eosinophilia and substantially increased IL-10 and the number of CD4+CD25+ Forkhead winged helix transcription factor box P3 (Foxp3+) IL-10+ T cells in the lung. Administration of PC61 antibody blocked the effect of Flt3-L and substantially increased AHR, eosinophilia, and BALF IL-5 and IFN-γ levels, and decreased BALF IL-10 levels and the number of CD4+CD25+Foxp3+IL-10+ T cells. Flt3-L significantly decreased CD62-L, but increased inducible costimulatory molecule and Foxp3 mRNA expression in the CD4+CD25+ T cells isolated from lungs of Flt3-L–treated, CRA-sensitized mice compared to CRA-sensitized mice without Flt3-L treatment and PBS control group. Flt3-L significantly inhibited the effect of CRA sensitization and challenge to increase GATA3 expression in lung CD4+CD25+ T cells. Collectively, these data suggest that the therapeutic effect of Flt3-L is mediated by increased density of naturally occurring CD4+CD25+Foxp3+IL-10+ICOS+ T-regulatory cells in the lung. Flt3-L could be a therapeutic strategy for the management and prevention of allergic asthma.
airway hyperresponsiveness; anti-CD25 antibody; Fms-like tyrosine kinase 3 ligand; Forkhead winged helix transcription factor box P3; naturally occurring CD4+CD25+ T-regulatory cells
Rationale: T-regulatory cells (Tregs) are potent immunomodulators in allergic asthma.
Objectives: We evaluated the functional effects of Tregs by adoptively transferring naturally occurring CD4+CD25+ Tregs (NTregs) and CD4+CD25− inducible Tregs (iTregs) from lung and spleens of green fluorescent protein (GFP)-transgenic Balb/c mice into cockroach-sensitized and -challenged mice.
Methods: GFP-labeled NTregs and iTregs were adoptively transferred into cockroach-sensitized and -challenged mice. Airway hyperresponsiveness (AHR) to methacholine was examined using a single-chamber, whole-body plethysmograph and invasive tracheostomy.
Measurements and Main Results: Adoptive transfer of either NTregs or iTregs from lung or spleen reversed airway inflammation and AHR to methacholine, and the effect lasted for at least 4 weeks. GFP-labeled iTregs up-regulated CD25 and forkhead-winged transcriptional factor box protein 3 and migrated to lymph node and lung. Lung CD4+CD25+ T cells isolated from each group of recipient mice were inducible costimulatory molecule–high and programmed death (PD)-1–positive; however, higher expression of PD-1 was found in the spleen iTregs (S25−) and lung iTregs (L25−) groups. Higher levels of transforming growth factor–β and IL-10 mRNA transcripts and bronchoalveolar lavage fluid IL-10 and INF-γ levels were observed in lung CD4+CD25+ cells from the L25− and S25− cell-recipient mice than from lung NTregs (L25+) and spleen NTregs (S25+) cell-recipient mice. Adoptive transfer of either cell type significantly reduced bronchoalveolar lavage fluid IL-4, IL-5, and IL-13 levels.
Conclusions: Tregs reverse AHR and airway inflammation; however iTregs that differentiated into IL-10–producing CD4+ type 1 cells in the lung exert their suppressive activity likely by higher levels of transforming growth factor–β, IL-10, IFN-γ, and elevated levels of PD-1 compared with NTregs. Hence, PD-1 may be a conduit for reversing AHR by Tregs and a plausible target for treating asthma.
airway hyperresponsiveness; forkhead winged helix transcription factor box P3; inducible CD4+ CD25− T-regulatory cells; naturally occurring CD4+CD25+ T-regulatory cells; programmed death-1
Dendritic cell subsets display different functional role in regulating immune response and lead to various outcomes including Th1 versus Th2 or regulatory versus immunologic response. Administration of Flt3-Ligand prevents and reverses allergic airway inflammation and airway hyperresponsiveness in a mouse model. However, the underlying mechanisms are unclear.
We characterized and examined the role of lung dendritic cell subsets in the therapeutic effect of Flt3-Ligand.
Dendritic cells were isolated from the lungs of OVA-sensitized and challenged mice treated with rhFlt3-Ligand. Two populations of CD11c+ cells labeled with fluorochrome-conjugated antibodies were sorted. The ability of the purified cells to stimulate T cell proliferation and cytokine secretion pattern by different DC subsets was examined. Also, dendritic cells were adoptively transferred in mice to examine their effect on pulmonary function.
Two dendritic cell populations, CD11chighCD11blow and CD11clowCD11bhigh, were identified in the lungs of naïve and OVA-sensitized and challenged mice with and without treatment with Flt3-Ligand. The expression levels of CD8α, B220, CD19, F4/80, MHC II, CCR7, CD40, PDL1, PDL2, CD80, and CD86 were distinctly different between the two DC populations, which supports the notion that, CD11chighCD11blow and CD11clowCD11bhigh dendritic cells, potentially have regulatory and immunogenic properties, respectively. Administration of Flt3-Ligand increased the dendritic cells with regulatory potential in the lungs of antigen-sensitized mice, and CD11chighCD11blow dendritic cells acquired a maximum degree of regulatory capacity after Flt3-Ligand treatment.
These data suggest that Flt3-Ligand reverses airway hyperresponsiveness by regulating the function of lung dendritic cells in a mouse model of allergic airway inflammation.
Flt3-Ligand could be a potential immunomodulator in the treatment of established asthma.
Adoptive transfer; Airway hyperresponsiveness; Asthma; Dendritic cells; Flt3-Ligand; Mouse model; T cell Proliferation; Regulatory dendritic cells
Vascular remodeling and atheromatous lesion formation are determined in part by the balance between apoptosis and survival of vascular smooth muscle cells (VSMCs). In the chronic stages, apoptosis of VSMCs in the atherosclerotic plaques contributes to the weakening and potential rupture of the plaque causing pathologies such as acute coronary syndrome. The higher incidence of apoptosis in the plaques of symptomatic than in asymptomatic patients has been demonstrated, but the expression of survival proteins, including the inhibitor of apoptosis proteins (IAPs), has not been thoroughly examined. The aim of this study was to investigate the immunohistochemical expression of cellular inhibitor of apoptosis protein-2 (cIAP2), x-linked inhibitor of apoptosis protein (XIAP), and survivin in normal carotid arteries, and carotid endarterectomy specimens of symptomatic and asymptomatic patients with carotid stenosis. The results demonstrated stronger immunopositivity to smooth muscle myosin heavy chain antigen (SM-MHC) (sm2), proliferating cell nuclear antigen (PCNA), and p50 subunit of NF-κβ in the asymptomatic plaques than in symptomatic plaques. Furthermore, there was higher expression of cIAP2, XIAP, and survivin in the symptomatic than in the asymptomatic plaques and this paralleled caspase-3 expression. The increased expression of IAPs in symptomatic plaques could be due to endogenous defense mechanism to protect against the pro-apoptotic effect of the inflammatory stimuli that are released in the plaques. This could be involved in the stabilization of symptomatic atheromatous plaques and may prove a potential therapeutic target.
Apoptosis; Atherosclerosis; Carotid plaque; Carotid endarterectomy; Inhibitors of apoptosis; PCNA; Vascular smooth muscle cell
Dendritic cells (DCs) are unique antigen presenting cells that are immature prior to their encounter with an antigen. Exposure to allergens induces the maturation of DCs with changes in morphology and presence of dendrites. Here, we demonstrate that the DCs in the lungs of ovalbumin (OVA)-sensitized and challenged mice are more mature owing to their pronounced dendrites than the DCs in the lungs and spleen of PBS-treated mice, which are immature and possess cytoplasmic veils. Intermediate to these two groups are the DCs in the Flt3 ligand-treated group that exhibit comparatively fewer dendrites and cytoplasmic veils and hence are classified as semimature. Presence of large numbers of well developed mitochondria and rough endoplasmic reticulum in myeloid DCs from both lungs and spleen of OVA-sensitized and challenged mice indicate greater functional activity. Additionally, DCs from the OVA-sensitized and challenged mice also exhibit fat and glycogen stores, which are indicative of a mature population. In addition, treatment of the animals with Flt3 ligand attenuated airway hyperresponsiveness to methacholine in OVA-sensitized and challenged mice. These data suggest that morphological features could be indicative of the maturation and distinct functional state of DCs, and this could be associated with underlying mechanisms of Flt3 ligand-induced immunomodulation in allergic asthma.
Airway hyperresponsiveness; Allergy; Asthma; Cytoplasmic veils; Dendritic cells; Flt3 ligand; Immunomodulation; Morphology of dendritic cells
Ceramide, a sphingosine-based lipid molecule, has emerged as a key regulator of a wide spectrum of biological processes such as cellular differentiation, proliferation, apoptosis and senescence. Sphingomyelinase-dependent hydrolysis of sphingomyelin, and de novo synthesis involving the coordinated action of serinepalmitoyl transferase and ceramide synthase, are the two major pathways involved in ceramide synthesis. Clustering of plasma membrane rafts into ceramide enriched platforms serves as an important transmembrane signaling mechanism for cell surface receptors. Ceramides have been implicated in apoptosis, stress-signaling cascades as well as ion channels. There is accumulating evidence that targeted manipulation of ceramide metabolism pathway has immense therapeutic potential and may eventually prove to be a boon in the design of novel strategies and development of innovative treatments for diverse conditions including cardiovascular diseases, cancer and Alzheimer’s disease. As yet uncharacterized natural ceramide analogs and novel inhibitors of ceramide metabolism might prove to be potent drugs. In this review, we discuss significant advances that continue to provide intriguing insights into the complex cellular and molecular mechanisms underlying ceramide-mediated signaling cascades.
Apoptosis; Atherosclerosis; Cancer; Cardiovascular diseases; Ceramide; Ion channels; Neurodegeneration; Raft; Sphingomyelinase
The existence of Cl- channels in vascular smooth muscle cells (VSMCs) has been increasingly investigated, but the biological functions are not yet clear. Insulin-like growth factor (IGF)-I affects proliferation and migration of VSMCs, and dysregulation of this axis may be involved in atherogenesis and intimal hyperplasia. We examined the effects of Cl- channel blockers on IGF-I-induced proliferation in porcine VSMCs. The siRNA approach was used to support the role of ClC-2, a member of the volume-regulated Cl- channel family, in cell proliferation of VSMCs.
Method and Results
The IGF-I-induced VSMC proliferation was significantly suppressed by the Cl- channel blockers NPPB and IAA94 but not by DIDS. IGF-I-induced cell proliferation parallel a significant increase in the endogenous expression of ClC-2 mRNA and protein. Inhibitors of PI 3-kinase, LY294002 and wortmannin, significantly attenuated the IGF-I-upregulated ClC-2 expression and cell proliferation. We observed ClC-2-like Cl- current, and this current was augmented by IGF-I. SiRNA specifically targeted to ClC-2 abolished IGF-I-induced cell proliferation.
Our data demonstrate that ClC-2 plays a role in IGF-1-induced regulation of VSMC proliferation in cardiovascular diseases.
Atherosclerosis; voltage-gated Cl- channels; Cl- channel blocker; Proliferation; Vascular Smooth Muscle Cells
The POK family of proteins plays an important role in not only embryonic development and cell differentiation, but also in oncogenesis. Leukemia/lymphoma-related factor (LRF) belongs to the POK family of transcriptional repressors and is also known as POK erythroid myeloid ontogenic factor (POKEMON), which binds to short transcripts of HIV-1 (FBI-1) and TTF-1 interacting peptide (TIP21). Its oncogenic role is known only in lymphoma, non-small cell lung carcinoma, and malignant gliomas. The functional expression of LRF in human breast carcinoma has not yet been confirmed. The aim of this study was to investigate and compare the expression of LRF in human breast cancer tissues and other human tumors. The expression of LRF mRNA transcripts and protein was observed in twenty human benign and malignant breast biopsy tissues. Expression of LRF was observed in several formalin-fixed tissues by immunohistochemistry and immunofluorescence. All malignant breast tissues expressed mRNA transcripts and protein for LRF. However, 40% and 15% benign breast biopsy tissues expressed LRF mRNA transcripts and protein, respectively. The overall expression of LRF mRNA transcripts and total protein was significantly more in malignant breast tissues than the benign breast tissues. LRF expression was also observed in the nuclei of human colon, renal, lung, hepatocellular carcinomas and thymoma tumor cells. In general, a significantly higher expression of LRF was seen in malignant tissues than in the corresponding benign or normal tissue. Further studies are warranted to determine the malignant role of LRF in human breast carcinoma.
Breast Cancer; Pokemon; LRF; ZBTB7; POK
Advances have been made in defining the mechanisms for the control of allergic airway inflammation in response to inhaled antigens. Several genes, including ADAM33, DPP10, PHF11, GPRA, TIM-1, PDE4D, OPN3, and ORMDL3 have been implicated in the pathogenesis and susceptibility to atopy and asthma. Growing evidence associates asthma with a systemic propensity for allergic type 2 T-cell cytokines. Disordered coagulation and fibrinolysis also exacerbate asthma symptoms. Balance among functionally distinct dendritic cell subsets contribute to the outcome of T cell-mediated immunity. Allergen-specific T-regulatory cells play a pivotal role of in the development of tolerance to allergens and immune suppression. Major emphasis on immunotherapy for asthma during the past decade has been to direct the immune response to a type 1 response or immune tolerance. In this review article, we discuss the current information on the pathogenesis of allergic airway inflammation and potential immunotherapy, which could be beneficial in the treatment of airway inflammation, allergy, and asthma.

domain: medical


text:
Originally Posted by mibs
I'm on my 3rd time playing Skyrim. My PS3 died during my first game, had to wipe it and start over, died again and had to send it in. I'm playing the same kind of character, but I haven't touched the main quest yet. I guess the game is all what you make it and the kind of story you build yourself. My character is a heavy drinker this play through because by this point she's seen all this craziness play out 3 times. My house is full of mead and ale and no furniture. Lydia keeps my secret. lol
The Companions are my favorite faction, felt so bad about the whole Kodlack thing. Dark Brotherhood is fun too, but I liked it better in Oblivion. Thieves guild is pretty awesome as well, I like the Nightingales aspect and that armor is BA.
Haha yeah, Im just the opposite, I done the main quest and started the side. Not the spoil anything for you but at the end you get a pretty AWESOME should which allows you to have some help in whatever you want (not saying exactly what /who it is I don't want to ruin things for you! )

domain: hobby


text:
Habitat is partnering with Cville Craft Aid to help make and distribute thousands of masks that may help protect against the transmission of COVID-19.
While a shortage of protective masks for health care workers has dominated the news, the Centers for Disease Control (CDC) now recommends that anyone venturing out into public places wear a mask that covers their mouth and nose. Since commercially manufactured masks have long since disappeared from retail shelves and online platforms, the only masks available to most people are homemade.
This shortage prompted Cville Craft Aid to organize local community members with sewing skills to pitch in and start making masks for people in need. The patterns for the masks are based on recommendations by the CDC. Habitat's primary role in the partnership is to safely assemble and distribute mask-making kits containing the necessary cloth, thread and elastic.
Distributing protective gear is not typically part of Habitat's mission, but our organization has expanded its scope in response to the pandemic. An interim plan to address the crisis was recently approved by Habitat's Board of Directors.
"Habitat is an organization that runs to the fire, so pivoting our operations to address a need in the community where we can make a big impact is part of our DNA," says Caitlin Riopel, Director of Operations. "Supporting mask making is definitely a new endeavor for us and we've learned a lot from the community experts who have been doing this work since the onset of the COVID-19 crisis."
As of September 2020 the mask program partnership with CCA has subsided as masks became more available.

domain: sociology


Manchester United are reportedly set to withdraw their interest in Borussia Dortmund and England attacker Jadon Sancho.
As per a report from the Daily Mail, Manchester United have decided to call off their pursuit of highly-rated England international Jadon Sancho, who has had a splendid 2018-19 campaign with Bundesliga outfit Borussia Dortmund.
A product of the Manchester City academy, Sancho joined Dortmund in the summer of 2017 in search of regular first-team football and has been a revelation in his second season at the club. So far this term, the 19-year-old sensation has come up with returns of 9 goals and 18 assists from 36 matches in all competitions.
Sancho’s displays for the German heavyweights resulted in him making his debut for the England national team in October last year. Till date, the winger has accumulated four caps for the Three Lions and is likely to be a part of the squad for the UEFA Nations League finals coming up in the summer.
The rate at which Sancho has progressed over the past year or so has seen him gather a lot of admirers across Europe, none bigger than Manchester United. The Red Devils had been heavily linked with the teenage sensation, who is valued around the £100 million mark by Dortmund, in recent months.
Manager Ole Gunnar Solskjaer, whose full-time appointment was confirmed last month, is keen on strengthening his attack heading into next season and sees the right wing as an area that needs to be reinforced. To that end, the 20-time English champions had earmarked Sancho as the ideal target and were set to launch a move for the Englishman.
However, as per the report Manchester United are ready to shelve their plans to sign the 19-year-old in the summer. The main reason behind the Red Devils’ decision is believed to be Dortmund’s unwillingness to do business as they have no intention of losing their crown jewel.
Borussia Dortmund director Michael Zorc had already issued a hands-off warning to all clubs monitoring Sancho, claiming ‘it will be very difficult even for the biggest of clubs’ to sign him, especially with the club already losing Christian Pulisic, who will be joining Chelsea at the end of the season.
Given that Sancho is contracted with Borussia Dortmund till the summer of 2022, the German heavyweights are in a strong negotiating position. Moreover, with the player also reportedly not seeking a move away at the time, Manchester United have accepted the fact that prising him away in the upcoming summer window will be a difficult task.
With the pursuit for Sancho set to be called off, Manchester United will have to look elsewhere to bolster their attack. There had been links to Chelsea’s Callum Hudson-Odoi, but that could also prove to be a dead end as per the report, while a move for former star Wilfried Zaha, who has been starring for Crystal Palace, has been mooted recently.


Low-key standouts Keithley, Hibner find high-energy youngsters all around them
By PAUL STURM, C-T Sports Editor
CHILLICOTHE, Mo. — When the Chillicothe High School Class of 2017 graduated, relegating the talented, large group of girls’ basketball players to the ranks of alumnae, some expected Lady Hornets’ basketball to slip back into the ranks of so-so programs, probably having winning seasons, but producing not much more than slightly-above-.500 season records.
Those who thought that have been proven wrong, with the two editions of Lady Hornets hoopsters since then producing a composite 37-17 mark and last year’s veterans-laden club posting 20 Ws and being denied a state-tournament appearance largely because it was in the same district as eventual state third-place finisher Trenton.
With three full-time starters – including all-district honoree and now junior-college player Kennedie Kieffer – and another occasional starter/top reserve from that club graduated and three other reserves who could have contributed to the 2019-20 squad – one a junior – opting not to play the sport this winter, some might anticipate the CHS girls losing some traction this winter.
Given the recent past, it would not seem prudent to count on it.
“We’re going to be fairly young – a freshman that’s going to play quite a bit, sophomores that are going to play quite a bit, and a couple of juniors that are going to play quite a bit, so the experience on the floor isn’t tremendously high, but, with the leadership I think we’ve got, I think we’ll be okay,” Smith shares in glancing ahead to the new season which is due to begin this coming Tuesday with a home game against Hamilton: Penney.
“We’re going to be more of a ‘5-out,’ motion-type (offensive approach). The big thing we’ve got to get to where we’re doing is not standing (still). Make the other team chase us, because we’re athletic enough and quick enough that we’ll be difficult to guard.”
“We’re just athletic and quick and long,” Smith further expands. “… If we need to ‘bang’ (play more physically) a little bit, (5’11” junior) Brooke Horton comes in. She did a good job, I thought, of ‘banging’ (in last Monday’s public intrasquad scrimmage) and showed her versatility, bringing the ball up the court. I think I can put five out at any time that can run the floor and bring the ball up the court and shoot the ball well.”
Despite the graduation and other personnel losses, this season’s basketball Lady Hornets will have two veteran, senior foundation piers to build upon with a half-dozen or so younger players who saw quite a bit of playing time a year ago and a promising freshman, as well.
Versatile guard/forward Hunter Keithley and record-setting shooting guard Jordan Hibner will be the bedrocks of the ’19-’20 CHS club – the former a multi-dimensional third-year starter who played extensively as a reserve late in her freshman year and the latter a perimeter sniper who broke the program’s quarter-century-old record for 3-points baskets made last season.
“Yeah, absolutely,” Smith unhesitatingly replied when asked if having two all-district and all-Midland Empire Conference players to fill in around puts his team in an advantageous spot, “and they’re leading this team. In practice, they’re doing all of the little things that we want to show these younger girls.”
As a junior on a well-balanced squad, the 5’10” Keithley led CHS in scoring a dozen occasions last year, averaging a team-best 11.8 points per game while shooting 80 percent from the foul line and 42 percent outside the arc. At different times, she hurt opponents with outside shooting, driving scores, and even posting up, in addition to the precise free-throw marksmanship.
Hibner was the Lady Hornets’ top scorer in six of their seven games last year, including draining a CHS single-game record seven treys and netting a team season-best 25 points in a mid-December victory over what became an outstanding Marshall squad, before foes got hip to knowing they could not give her room to shoot from deep. She added a seventh occasion of being CHS’ leading scorer in late January and, with her final basket of the season, knocked down her 71st triple of the year, eclipsing the former Lady Hornets’ standard established by Rochelle Barrett in 1994-95. Hibner finished with a scoring average of 9.2 points and led the club with 1.3 steals a game and 62 percent adjusted field-goal percentage (43 percent outside the 3-points line).
The biggest question mark about that duo – particularly Keithley – is whether they will be selfish enough to help their younger teammates.
“That’ll be key for us, to be honest,” Smith candidly confirms, “is they’re going to have to look for theirs (shots and scoring chances).”
However, the duo’s overall skill, experience, and willingness to help their teammates flourish makes any small worry about their being ready to shoulder extra load early is minimal.
“We’re a long way from being where we need to be, but just seeing the floor the way they do is going to help us get the other girls the ball in the right spots” to prosper, the coach says of the pair of proven senior standouts.
While, assuming good health, the low-key Hibner-Keithley tandem’s contributions figure to be a given, what seems poised to put the “juice” into this year’s – and upcoming years’ – CHS girls’ squad is an array of quick, aggressive players in a variety of sizes, several of which more than got their feet wet last season.
Most-experienced of that group will be lanky sophomore forward Essie Hicks and energetic, aggressive point guard Lucy Reeter, another 10th grader who looks likely to see time at both the “1” (point) and “2” (shooting) guard spots. Both got into more than 20 games a year ago.
Hicks looks to have lost any timidity about her game with that prior experience and, with her height, long arms, and speed, could create mobility mismatches with other teams’ post players, as well as being a harassing presence at the top of the press the Lady Hornets figure to use very extensively.
“Essie’s really come along,” Smith praised. “She’s just so deceivingly fast. She just glides.
“We’re looking to get her down the court fast to try to feed it to her (in transition). Her job is supposed to be to beat the other team’s ‘big’ down the floor and she’s going to be able to do that a lot.
“She’s rebounding the ball well. Her head’s in the right place. She was getting us right into our press right away every time (during the intrasquad scrimmage). Really did a good job.”
Quick and fearless on the drive, but with good ballhandling ‘chops,” L. Reeter will bring a dribble-drive dimension CHS hasn’t had since Karson Keithley’s graduation in 2017, but she won’t be alone in reintroducing that capacity.
Classmate Selby Miller will bring quickness and an attacking mentality to the point, as will freshman Jessica Reeter, who not only is expected to see lots of action, but could start at the outset of the season next week.
“She just doesn’t seem to get tired,” Smith says of the frosh, already adept at using either hand for dribbling or passing.
Of the sophomore pair, the CHS coach reports, “Both of them can run the ‘1’ or the ‘2’. Both of them can shoot the ball well and dribble-penetrate. Selby might be a little more defensive-minded than Lucy, but, man, Lucy can handle the ball and get it where it needs to go, so they play off of each other real well.”
He continues, “It’s just going to be a matchup, trying to figure out who’s working well together for us. In the long run, I’ll be able to throw about anybody out there in that group.”
In addition to that young trio, senior Haley Kidd, who showed nice potential as a transfer from Trenton a year ago, seems to have found her niche and comfort zone as a “2” (shooting) guard who won’t be afraid to “drive and dish.”
“She plays hard,” compliments the coach. “Coming where she came from at Trenton, where they played mainly zone (defense) to, pretty much, straight ‘man’ (under Smith), I feel like her principles are real good.
“She’s coming along really nice.”
That quartet will be expected to create more need for “help” defense from the players primarily assigned to guard Hibner and Keithley on the perimeter. That should translate to more clean looks at the rim for that sniping pair, as well as for Keithley to cut to the basket unguarded for shots and foul-drawing attempts when her defender gets distracted by the point guard’s penetration.
“That’s kind of one of the reasons why we changed what we’re doing. No. 1 is the players fit it a little better. You’ve got Lucy and Selby and Jessi – any one of the three can run that ‘1’ (point guard position) and break people down off the dribble. That’s going to open my (outside) shooters, I think, a lot.”
Based on who Smith sent out to begin this past Tuesday’s “jamboree” scrimmages against Brookfield and Trenton, the starting lineup for Tuesday’s opener against a good Hamilton team, led by star guard Nora Ford and others, projects to be Keithley, Hibner, Hicks, L. Reeter, and Je. Reeter. Horton, Miller, and Kidd would be the earliest reserves to enter.
Trenton star Maci Moore abused CHS in the paint on both offense and defense in the “jamboree” scrimmage and developing a more-hard-nosed attitude overall and especially about rebounding and defending inside looks to be one of the primary tasks – along with sharpening the shooting eyes of everyone – to be undertaken during the 2019 portion of the season.
Finding post-player depth will be another key mission during December and early January. One or more from the group of junior Ella Leamer and sophomores Clara Leamer and Ellie Barnett will be needed to provide foul-trouble protection, as well as personnel capable of helping keep the press in use even when starters are resting. If any of those three can show the moxie to battle defensively and for rebounds under the glass, that player likely would earn the most playing time.
“That’s where we’re not real big. … You’ve got (5’9”) Clara, (5’6”) Ella, and (5’7”) Ellie that are going to have to run through that (low) post. Their job’s just going to be to battle, just be strong, be physical, and they’re pretty good at that. They work really hard.”
As was demonstrated during last Tuesday’s “jamboree” scrimmages against talented and experienced Trenton and Brookfield teams which both appear headed to successful seasons – BHS under Chillicothe alumnus Tony Fairchild and THS under Kameron Cool, son of CHS boys’ head coach Tim Cool, there undoubtedly will be growing pains for the Lady Hornets, given the turnover in starting personnel. The raw materials to work with, though, clearly seem there.
“Those younger kids just have to learn the speed of the game. We’ve been practicing at that speed, … so they’re starting to get a grasp of what that’s all about. But, man, they play hard and that’s what we want.
“We want them to play hard and go (full speed) for two or three minutes and then we’ll get somebody else in there” to maintain the fast pace.


Envelope-Designer is a plugIn with which you can adjust the relative amount of the attack transients and the sutained/release components in a signal. For the detection of the transient component there are 2 envelope-followers with identical release times and different attack times. The difference between the envelope with the fast attack time and the envelope with the slow attack time yields a control signal, which has significant amplitudes only when the incoming signal is in some "transient" state. This is because the difference between the envelope-followers with fast and slow attack tends to zero in a sustained phase.
Multiplying this control signal with the incoming signal yields a signal, which contains the transient component. The extraction of the release component is similar: it is done with two envelope followers with identical attack times a different release times. The difference between the envelope with the slow release time and the envelope with the fast release time has significant amplitudes for decaying signals. Multiplied with the incoming signal, we have extracted the release component. Now we can adjust the relative amplitudes of these two signal components seperately.


text:
Once you hear the term International Federation of BodyBuilding and Fitness or IFBB, you in all probability consider huge bodybuilders on the stage flexing. Do place a land weight training poster in your swimming pool, do place a swim fitness poster in your weight room. Train posters serve to advertise fitness, start a dialogue together with your private coach, encourage you to rent a personal trainer and most of all encourage you to stick together with your fitness program. The routines are straightforward to comply with, which is great for those new to Zumba® fitness or both of the packages, Gold® or Firming.
Be taught in regards to the elements of bodily fitness: cardiorespiratory endurance, muscle strength, muscle endurance, flexibility and body composition and why they’re essential. Although Sarah Bäckman isn’t a fitness model or bodybuilder…you can’t say she isn’t match! Female Fitness Competitor Sylvia Tremblay has a ton of fitness competitions under her belt. GoodLife Fitness Instructors do greater than train a Fitness Class — they motivate Members to maximize their workout, exceed their fitness objectives, and type relationships in a fun group setting. This beautiful younger woman (that jogs my memory a bit of former WWE Diva Michelle McCool for some motive) is fitness mannequin and female fitness coach Aubrie Richeson. The Class Starter helps match you with courses that fit your private fitness objectives and schedule.
Vixen Exercise’s intense Hip-Hop cardio has taken the dance fitness scene by storm. Other than our military fitness periods at venues nationwide, we additionally supply mud run events, a fitness vacation, corporate packages and fitness qualification courses. Gail competed in the 2004 Fitness Success Challenge and the 2004 Womens Tri-Fitness in Las Vegas. Filipina IFBB determine competitor, fitness model and certified private coach Meriza DeGuzman Ciccone is proof that good issues come in small packages. I believe Jennifer Nicole Lee is probably one of the vital stunning fitness models on the market.
Kim Kanner was also a fitness model, but actually wasn’t much into the wrestling facet of the business. Remember, fitness contains equal portions of: 1.) goal toning 2.) cardiovascular train three.) flexibility (stretching) four.) food plan and 5.) consistency. Lithuanian fitness mannequin and bodybuilder Živilė Raudonienė left a really promising IFBB Figure profession to go to work for the WWE as Aksana. At VI Fitness we aim to provide you with the assist and encouragement you should obtain your health and fitness goals! Exotic fitness magnificence Dana Hamm has appeared in fitness magazines like Muscle Magazine, Planet Muscle and Muscular Improvement. So I started seeing a fitness trainer, I attended Planet fitness which is only 10 dollars a month so its very inexpensive and I’m set up with a coach who don’t value me a dime!!
Our self-guided exercise apps have helped over 1 million individuals work towards reaching a variety fitness goals, from weight reduction to muscle achieve, flexibility to harm restoration. These feminine models look great, it takes a large amount of self-discipline to achieve that stage of physique.

domain: sports


text:
(1) A person whose DNA profile has been included in the database and databank pursuant to this chapter may make a written request for expungement of materials from the database and databank on the grounds that the conviction upon which the authority for including the DNA profile was based has been reversed and the case dismissed.
Terms Used In Idaho Code 19-5513
- Appeal: A request made after a trial, asking another court (usually the court of appeals) to decide whether the trial was conducted properly. To make such a request is "to appeal" or "to take an appeal." One who appeals is called the appellant.
- Conviction: A judgement of guilt against a criminal defendant.
- Defendant: In a civil suit, the person complained against; in a criminal case, the person accused of the crime.
- Evidence: Information presented in testimony or in documents that is used to persuade the fact finder (judge or jury) to decide the case for one side or the other.
- person: includes a corporation as well as a natural person;
Idaho Code 73-114
State: when applied to the different parts of the United States, includes the District of Columbia and the territories; and the words "United States" may include the District of Columbia and territories. See Idaho Code 73-114 Trial: A hearing that takes place when the defendant pleads "not guilty" and witnesses are required to come to court to give evidence.
(2) The person requesting expungement must send a copy of his request, with proof of service on all parties to the following: the trial court which entered the conviction or rendered disposition in the case; the bureau of forensic services; and the prosecuting attorney of the county in which he was convicted. The court has the discretion to grant or deny the request for expungement. A trial court’s denial of a request for expungement is an order not subject to appeal.
(3) Except as provided below, the Idaho state police shall expunge the DNA sample and all identifiable information in the database and databank relating to the subject of the conviction upon receipt of a court order which verifies that the applicant has made the necessary showing at a noticed hearing, and which includes the following documents:
(a) Written request for expungement pursuant to this section;
(b) A certified copy of the court order reversing and dismissing the conviction;
(c) Proof of written notice to the prosecuting attorney and the bureau of forensic services that such expungement is being sought; and
(d) A court order finding that no retrial or appeal of the case is pending and verifying that at least sixty (60) days have passed since the defendant has notified the prosecuting attorney and the bureau of forensic services of the expungement request and that the court finds no reason, based on the interests of justice, to deny expungement.
(4) Upon order of the court, the Idaho state police shall destroy the DNA sample relating to the subject of conviction, unless the state police determines that the person has otherwise become obligated to submit to DNA sample and thumbprint impression as a result of a separate conviction subject to the terms of this chapter.
(5) The bureau of forensic services is not required to destroy an item of physical evidence obtained from the DNA sample if evidence relating to another person subject to the provisions of this chapter would thereby be destroyed. Notwithstanding this subsection, no sample, physical evidence or identifiable information is affected by an order to set aside a conviction.

domain: law


text:
Tania AdamsLicensed Clinical Social Worker, MSW, LCSW
Are you struggling with boundaries, expectations that are self imposed, exhaustion and feeling "not enough"? If you are having difficulty navigating your relationships, parenting, big (or small) life transitions, there is a way for you to orient back to yourself so you can make decisions from a place of clarity and to find more connection and meaning in your life. Whether you're trying to connect better in your relationships, or with yourself, things can change.
Gig Harbor, WA 98335
0.0 Miles away

domain: psychology


text:
Use a chic and captivating layout such as this item to promote your branding and leave a lasting impression on your potential clients. Five shots of this elegant screw cap dark glass wine bottle that is illustrated in the dark foreground will provide you with a perfect chance to use your favorite sights.
This wine bottle mockup can be customized with no effort or charge. Set its features with a few clicks and after that, have a look at our inspiring gallery on the Resource Boy website. We will help you to make a lot of amazing artwork.

domain: design


We never believed our show would fill a stadium—P-Square
As kids they saw Michael Jackson as an Idol and started what will become their career, dancing and singing to the beats of Wacko Jacko. Never in their wildest dreams did they think of one day pull a crowd staging a show that will filling a stadium to full capacity, but, P-Square achieved that in their Africa Invasion tour. In this chat with AHAOMA KANU. The duo of Peter and Paul Okoye popularly known as P-Square share the magical moment when they took the stage they once watched their Idol do many times in many countries.
You have been on tour of the African Invasion, how has it been?
Well, I have to say that we have been top 70 per cent of the African continent and I will tell you that the first experience was when we went to Sierra Leone. When I was informed that we will be going there I asked, is our music there but it turned out to be the biggest concert we have been to in our Invasion Africa Project. We had like 62 000 fans coming to the show. You can imagine coming out from the plane and everybody was coming out but we were told that we had to wait and we didn't know what was happening. I peeped out from the window and saw the crowd waiting for us. Before we knew it, there was a red carpet and their cultural troupe came out dancing and welcoming us. I saw a man pointing to us to come down. The show promoter told us that the man there was the PA to the President. We walked on and then he told us then he pointed out that the president was waiting for us. We were surprised and really privileged. We shook hands with him and couldn't believe it.
Can you mention some of the African countries that you have been to and the ones you are yet to go visit as I am informed that you are booked out?
We cannot be able to count that now but I can remember Liberia, Zambia, Uganda twice, Gabon Twice, Ghana, Guinea, Sierra Leone, Rwanda and so on.
How did you feel then that a president of a country was present to welcome you at the airport?
Like I said, it was the first experience for us and we were like this is the Michael Jackson treatment. The president took us to the government house and we met some govt officials and then went to TV stations and radio stations before we now went to a dinner with the president and government officials. The next day we went for the media parley and were told us that the concert will be in a stadium; we were thinking that they were referring to a basketball stadium or so without having the least idea that they were using the Freetown International Stadium. We went there and were told that the stadium was filled up. We again thought that maybe it was filled with 5000 or 10 000 people but when we came out, they stadium was filled to capacity. I have to tell you the truth. It shows how important and highly regarded we are; it proves that what we are offering is highly accepted and regarded by fans across the African continent.
How did you feel seeing that P-Square drew a crowd that filled a stadium?
To be sincere, I could not believe that the people came to see P-Square. I thought that maybe they came to watch an international match.
Have you ever thought in your life you will fill a stadium?
My brother we never thought it would happen that way and it was surprising when it happened. When Jude saw the crowd he said we will increase our price. You see, they promoters wanted to use half of the stadium and put a backdrop but when the situation changed, they had to pull the backdrop down. We had the job of moving around the stage to be facing the whole crowd. It was fun.
Many foreign stars has had fans falling over and fainting at their shows and seeing such happening in the Invasion Africa preview was awesome, how did you feel seeing that playing out?
It's surprising and our Mum predicted that such a thing will happen to us. She predicted that even before we started the music thing; when we were still doing the dancing thing.
With that kind of acceptance you get outside the shores of Nigeria how do you compare it with the relative recognition you have here back home?
It's like the same; Nigeria is home and this is where we are from, where we live and Nigerian fans accept us very well; they love what we are doing because we thrive to satisfy them but the most surprising thing about the whole issue is that you don't know how your music gets to those places; we sit down here and we get calls that you are wanted here and there and no matter the bill you present, they are willing to pay. It is just as Nigerian brands bring in international artistes here. We can pay them heavily to make sure they come here, that is the same way those promoters in these countries want to have us there. So in comparison, it can't be the same, this is home. Again, when we travel out of the country, we don't look at the music aspect; we dwell on the culture, their system; we look at the country and the system and compare them with Nigeria. I must tell you that I am amazed at what we see there.
Can you tell us some of the surprises you have seen there compared to Nigeria?
Good road, Electricity; all the countries we have been to, there has not been power disruption for any single day. Even in Rwanda that just came out of a war not quite long ago having uninterrupted power supply. We were surprised that a country that came out of a war 14 years ago has gotten their acts right. Many of us have seen the movie Hotel Rwanda and know the story of what happened there; we have been to Uganda and we know the history of Idi Amin and what he did to the country, we have been to Liberia and other countries and you find some of these amenities working and then you feel sad about Nigeria. After these wars, these countries developed from the ruins and got better and here we are claiming that we are the giant of Africa when our power sector is in coma. We have the biggest oil wells in Africa and yet our government cannot provide us with power.
How do you feel on those moments?
I feel bad. When we see all these developments I don't look at the music; I don't look at our songs all our dances, we tend to look at the infrastructures and the government and feel very disappointed. Why is our country like this when we are not in any war, how come we are not developed as they are?
Are you thinking of singing about the experience in your future songs?
Watch out for it my brother; we are getting details, data and information. Come to think of it, the international acts that come here, how many of them have met the president? But we have met presidents of many African countries we have been to; we have met the president of Sierra Leone and that o Rwanda and also have been hosted by many government officials and ministers. Imagine a minister in Rwanda driving us around town and they are that simple. In Rwanda, the president will drive his kids around to school and all time. The minister brought us to the stadium and we sat close to the president seat and when he came everybody stood up and we were expecting to see security and all the kind of armed details we have here but nobody was lurking around the president. We will definitely put these into our next album. We want our government to provide the basic infrastructures for the people and there is no alternative otherwise we will keep singing it till they do it.


Dangerous intersection to be studied
EFFINGHAM CO. (WJCL) Efforts are underway in Effingham County to make a dangerous intersection safer.
Tuesday night, commissioners paved the way for a feasibility study at Blue Jay Road and Highway 17.
The intersection of Blue Jay Road and Highway 17 in Effingham County has been a problem area for many years.
It's been the scene of serious accidents, including a few deadly ones.
Tuesday night, Effingham County commissioners agreed to pay for the electricity and maintenance of the area, thus allowing the Georgia Department of Transportation to begin a feasibility study to figure out how to make the intersection safer.
"They'll be determining based on all different kinds of scenarios whether it's a roundabout or a red light or what is most feasible there," said Vera Jones, Effingham County Commissioner. 'But us making the vote we made last night allowed them to go ahead and move forward on doing the study."
Most agree something needs to be done at the intersection, but according to one resident, a roundabout is not the answer.
"A traffic light, people in this county aren't going to know what to do with a roundabout," said Gloria Sheppard, Effingham Co. Resident. "Tthey're going to be confused and it's going to cause more wrecks, they should put a traffic light there."
"Everybody's concerned about that area, and want to see something done if possible," continued Vera Jones. "And that's the reason to go ahead and have the feasibility done, this is the starting point."
The bottom line is whatever work they decide to do at Blue Jay Road and Highway 17, it won't happen tomorrow. This is a long process that could take up to four to five years.
The feasibility study will begin sometime in the next two to four months.


Tastiera Piano Widget lets you use your computer as a piano. It gives a range of 3 octaves (1½ on either side of middle C).
It was originally made for singers, but should also be useful for composers, musicians, etc.
The widget adapts itself to any keyboard layout.
This is shareware with a 30-day trial period.
What's new in Tastiera Piano Widget
Fixed problems with non-US keyboard layouts.
Try our new feature and write a detailed review about Tastiera Piano Widget. All reviews will be posted soon.


text:
FROM Hogwarts to T in the Park, meet the Harry Potter star who has swapped her wand for a microphone to host this year’s festival coverage on BBC Scotland.
As a presenter on CITV’s Scrambled, Arielle Free is a favourite with young viewers but she’s hoping her T gig will make her a household name with audiences of all ages.
The 29-year-old, who grew up in Stirling, is one of TV’s rising stars and is being tipped as the next Edith Bowman or Jo Whiley.
Arielle, drivetime DJ for Ministry of Sound, is a massive music fan and can’t wait to be the first person to present from T in the Park’s new venue at Strathallan Castle in Perthshire.
The former dancer, who starred in Harry Potter and the Goblet of Fire, said: “I am getting excited about T. I was last there as a punter in 2007 and can’t wait to go back as a presenter.
“Music is my biggest passion and I’m looking forward to seeing some of the best bands in the world play live. From Kasabian, St Vincent, Mark Ronson and Sam Smith to Avicii, David Guetta and the Prodigy, it’s going to be amazing.
“I’m also looking forward to hearing The Proclaimers. It’s going to be a fabulous weekend and the new venue makes it extra special.
“Although my family are used to seeing me on TV, the fact I’m going out live on the BBC is making them very excited, especially my grandad, who gets up early at the weekends to watch me on Scrambled even though it’s just for kids.”
Arielle, who is hosting coverage of the West Halts stage at Glastonbury thisweekend, started out as a dancer and got into presenting by chance.
Represented by the same agent as Russell Brand, Caroline Flack and Dermot O’Leary, she attended the Dance School of Scotland at Glasgow’s Knightswood Secondary.
She was in her first year at dance and drama college in London when she got picked to play a Beauxbaton’s pupil in the fourth Harry Potter film.
Arielle, whose partner Ryan is also a huge music fan, said: “When I was younger, I always dreamed of being a dancer and was delighted when I got the part in The Goblet of Fire. It was an amazing job.
“After deferring uni for a year to film the movie, I went back and continued to dance. I appeared on Top of the Pops, CD:UK and even did backing dancing for Fatboy Slim.
“In 2006, I was dancing at the Reading festival and my friend asked me, ‘Can you just do this backstage interview for us since you chat a lot?’
“I was given a microphone and suddenly I had a camera shoved in my face and I loved every minute of it. I had been bitten by the presenting bug and knew that’s what I wanted to do.”
After Reading, Arielle got a job as a host on children’s TV show Pop Girl on Sky. She went on to present on 4Music for Channel 4 and then as a radio DJ for various stations before landing her current job at Ministry of Sound.
Arielle, who has interviewed everyone from Justin Bieber and Little Mix to Rizzle Kicks, would
love her own show on Radio 1.
She said: “Jo Whiley and Zoe Ball are my idols. I grew up listening to them and would love to follow in their footsteps and have my Radio 1 show.
“I also love Jools Holland. People say that music shows have died a death on TV but I don’t think they have. They just haven’t got the right concept yet. I’d love to do a sort of TFI Friday, mixed with a Jools Holland kind of vibe.”
Arielle believes people remember her because of her Scots accent and auburn hair. She said: “I’ve always loved my red hair and it has helped me land a few jobs.
“I’ve worked hard to get where I am and hope I’m in it for the long haul. If I could have a career with the longevity of Lorraine Kelly’s, I would be happy.”
See Arielle on BBC Scotland on Saturday, July 11, and Sunday, July 12.

domain: entertainment


text:
This is part of another project
An online drawing app that let's you change colors via MIDI Controller
The artwork on the cover of the controller case is designed with Blender and an addon called Sverchok. It let's you generate curves (in this case with the contour node) and export them to gcode. I attached the .blend file with the whole setup.
The different colors are printed with a diamond hotend.
List of parts:
- Arduino micro
- 3x 60mm linear fader (the ones with a green bottom slide the best, because they don't slide
on the resistor tracks itself, but on plastic rails above the tracks)
- male micro USB Jack (to connect Arduino to USB cable, easier to solder)
- 4-core headphone cable to make thin USB cable
- M2 screws (to screw bottom to top)
- rubber feet (flat gaskets)


Purpose: The purpose of this page is to provide a central place to store information on strain gage technology used in PSAS projects.
Strain gages are resistive devices which can be used to measure changes in length, or strain, of materials that occurs as forces are applied to those materials. They can be used on metals, plastics, and even seemingly inflexible materials such as concrete. A strain gage is mechanically bonded (glued) to the material under test so that any change in the length of the material will result in a change in the length of the strain gage. As the strain gage changes in length, there occurs a small change in the resistance of the strain gage. Measured carefully, the resistance change very accurately reflects the change in length of the material under test.
Background on strain gauges: http://www.omega.com/literature/transactions/volume3/strain.html
Strain gages we use:
...can measure a maximum of 30,000 microstrain for the SGT series http://www.omega.com/techref/pdf/STRAIN_GAGE_TECHNICAL_DATA.pdf
Definition of microstrain: http://www.unc.edu/~rowlett/units/dictM.html
One of the primary reasons we are using strain gages is for measuring tensile or compressive forces in devices called Load Cells. Our load cell designs can be found here.
Strain Gage Amplifiers:
Load cells require sensitive amplifier circuits to measure the tiny changes in resistance that is related to strain. Our amplifier designs can be found here (link).


text:
The first thing to make clear is that EU law has not been the sole – and perhaps not the primary – source of discrimination law generally, and sex discrimination law in particular. The Equal Pay Act 1970, one of the earliest modern anti-discrimination statutes, was provoked by the UK’s impending membership of what was then the EEC, and many subsequent amendments to UK sex discrimination law resulted from EU legislation or decisions of the CJEU (as it is now). But the Sex Discrimination Act 1975 was not driven by any EU imperative, having been preceded by Conservative proposals concerned to widen the range of opportunities open to women and to challenge unfair sex discrimination in areas such as employment and training. In its final form the Labour government’s 1975 Act owed more to the model which had developed under the Race Relations Acts of 1965 and 1968 and developments in the US in the 1970s than it did to Europe, although it became the vehicle in Britain for the transposition of the Equal Treatment Directive and was subject to many express amendments to this end. Neither that Directive nor any subsequent gender equality directive has matched the material reach of the 1975 Act which prohibited sex discrimination not only in employment (broadly defined to include vocational training and access to self-employment, trade union membership, etc) but extended, inter alia, to education, housing and the provision of goods, facilities and services.
Underpinning of rights
EU membership has, however, been extremely influential in relation to a number of matters, among them the right to equal pay for work of equal value and to protection from discrimination related to pregnancy, also to questions of enforcement. The ECJ (as it then was) required the amendment of the Equal Pay Act to cover equal pay for work of equal value, and interpreted sex discrimination to include discrimination related to gender reassignment. It prohibited differential retirement ages for men and women and the application of any cap on compensation for discrimination. A large number of cases developed the right to equal pay for work of equal value, and applied it to women working part-time. Others applied the prohibition on gender-related pay to occupational pensions and insisted that pregnancy discrimination amounted to sex discrimination.
These decisions all proved to be of substantial, and some of them of profound, importance to domestic law. Domestic discrimination law is now consolidated in Great Britain (though not Northern Ireland) into the Equality Act 2010 which regulates discrimination, victimisation and harassment because of sex, pregnancy and maternity and gender reassignment as well as race (broadly defined), sexual orientation, marriage and civil partnership, religion or belief, age and disability across a range of activities including employment, education, public functions, membership organisations and the provision of goods, facilities and services.
It is not inevitable, and indeed is unlikely, that Brexit will cause the edifice of domestic equality law to crumble. It is true, however, that Brexit will remove from domestic equality legislation the underpinning provided by EU law both in its current form and with the benefit of any future developments. This may well prove of increasing significance given the general hostility of many of those on the right to ‘red tape’ and the ‘compensation culture’. Certainly the prohibitions on discrimination in relation to pregnancy and maternity in particular impose ‘burdens on business’, as do restrictions on indirect discrimination related to sex.
Recent legal developments
Governments since 2010 have not introduced radical changes to substantive domestic equality law, though this may have been the result between 2010 and 2015 of the Liberal Democrats’ relative enthusiasm for equality legislation. A number of the innovations contained in 2010 Act have been scrapped, however, among them the regulation of ‘dual discrimination’ and the protection of workers from third party harassment (including sexual harassment). Mandatory gender pay audits have not been introduced, except in some cases following a finding of gender pay discrimination; in 2014 the UK for the first time, the World Economic Forum’s Global Gender Gap Report ranked the UK outside the world’s 20 most gender-equal countries (lagging behind Nicaragua, Rwanda, the Philippines, Bulgaria and Burundi as well Iceland, Finland, Norway, Sweden and Denmark) after women’s average wages fell by £2,700 in a year (men’s wages remaining constant).
The matters mentioned immediately above were beyond the scope of EU law and within domestic government’s freedom of manoeuvre. It is not fanciful to suggest that Brexit will result in other amendments to the scope of domestic law. Particularly significant as regards possible future directions of travel have been the changes made in recent years to the enforcement of equality/discrimination law. Discrimination/equal pay questionnaires, which had been in operation since the 1970s, were abolished as of April 2014 notwithstanding an 83% majority against the proposal in responses to the government’s consultation document. Also removed for apparently ideological reasons were employment tribunals’ powers to make recommendations, by way of response to successful discrimination claims, which extend beyond the relationship between claimant and employer. These powers were scrapped as an unnecessary burden on business despite an absence of any evidence (even assertion) of inappropriate use by tribunals, and notwithstanding the fact that only 12% of respondents to the consultation paper were in favour of repeal, with 79% against.
Budget slashing and impact of tribunal fees
Other assaults on enforcement have included the slashing of the EHRC’s budget from £70m in 2007 to under £18m in 2015/16, this despite warnings to then Home Secretary, Theresa May from the UN Commissioner on Human Rights in 2012 that the Commission would lose its financial autonomy, and its UN ‘A’ grade rating, if cuts to its budget continued. And perhaps most importantly, tribunal fees introduced in July 2013 resulted in a spectacular reduction of sex discrimination (and other) tribunal claims, discrimination claims now costing £1,200 to litigate and £1,600 to appeal. Fees may be remitted for claimants on very low incomes, but most claimants will be required to meet them as a condition of the claim. Tribunal claims immediately fell by 64%, sex discrimination and equal pay claims by 83% and 77% respectively. Notably, in view of the assertion by the-then Minister for Education and Business in April 2014 that the 79% drop in tribunal applications to that date demonstrated that blameless employers had previously been subject to many false allegations, the success rate of those claims brought has not changed in the period since fees were introduced.
The introduction of tribunal fees has clearly had a devastating impact on discrimination claims and it is strongly arguable, notwithstanding occasional expressions of support for gay marriage, equality and apple pie, that elements within the Conservative Party are constitutionally ill-disposed towards discrimination legislation. Brexit removes the counter-weight of EU law which has resulted in significant improvements for women in the UK, and which has no doubt prevented regressive steps which would otherwise have been taken. It is the case that attempts to rely on EU law to challenge tribunal fees have failed to date, the High Court (twice) and the Court of Appeal rejecting the argument that their imposition breached the EU principles of effectiveness (by effectively barring access to the court). But any suggestion that, in light of the tribunal fee challenges, that EU membership has proven ineffective in securing effective equality and related rights and that Brexit will make scant difference is premature; the Supreme Court’s decision on tribunal fees is awaited.
Early likely losses
Brexit is likely to do real damage to women. Even leaving aside specific discrimination law, women are disproportionately numbered among the most vulnerable workers, and are likely to be disproportionately affected by the expected bonfire of provisions such as entitlements to paid holiday and to maximum weekly hours and parental leave rights. Brexit also poses a specific threat to any elements of discrimination law which are seen as business-unfriendly. These include equal pay for work of equal value, as well as protection from pregnancy-related discrimination, which is rife. While it is perhaps unlikely that these rights will simply be abolished, the assault which has already been launched on their enforcement is likely to continue with early likely losses, including uncapped damages for discrimination claims and the recoverability of six years’ of back pay by successful equal pay claimants.
Contributor Aileen McColgan is Professor of Human Rights Law at King’s College London and a barrister at Matrix Chambers
SEX DISCRIMINATION RIGHTS REVIEW
Amid fears that Brexit will ‘turn the clock back’ on women’s rights, a review of sex discrimination laws was launched by the Fawcett Society. Its remit includes the effectiveness of current laws and how best to balance the rights of the individual with the responsibilities of the organisation.
Due to report in the autumn, the panel is led by retired High Court Justice, Dame Laura Cox DBE and coordinated by equality law expert Gay Moon. Panel members include a number of QCs and equality law experts. The closing date for evidence was 3 July but late submissions may be sent to Lawreview@Fawcettsociety.org.uk
#FaceHerFuture The Fawcett Society campaign to protect women’s rights post-Brexit is supported by over 20 women’s and equalities organisations.


text:
Alternative style of learning goes nonprofit
Herne Schools of Discovery, located on Youngs Lane, went nonprofit on March 3.
The school has been for-profit since it began in 1997, but owing to the need to raise more money through fund-raisers, the school filed for nonprofit status, or 501C3, which is a public or charitable purpose.
Unlike most schools that are accredited by the National Association of Education for Young Children, the Herne Schools of Discovery are not subsidized by the state, and as a for-profit organization, the donations to the school are not tax deductible.
According to Cherie Herne, head of the school, that can make it hard to solicit help.
The school needs accreditation from the NAEYC to become a quality care school, Herne said. This can be very expensive, as this covers insurance for the teachers, more vacation breaks, and, according to Herne, more education.
If the school does get the accreditation, the parents who enroll their children get double tax credit.
Another reason Herne needs to raise funds is because the school is contemplating moving to a new location on Route — by the York Sports Center. The new location would feature an Olympic-sized pool. The school also needs to raise funds for a school bus with safety restraints.
Herne said the school receives a lot of donations of material and funds through the community. That occurred even when the school was for-profit.
"It's really a beautiful thing," she said.
Herne said fund-raising also shows the school's commitment to its students and gains community camaraderie.
"It makes the staff proud," she said.
The alternative private school teaches preschool through second grade, and will add a grade each year until they go up to eighth.
Herne said the school tries to "teach core values" to its students. She says these would be how to treat other people and how to treat the Earth.
"It's a hands-on learning environment," she said.
The curriculum at the school is based on the curriculum of Berwick Academy and other private elementary schools. Each child's curriculum is different, however, according to his or her strengths or weaknesses.
"Everybody's working on their own individual guidelines," Herne said.
The school uses its discretion on when to enter children into kindergarten. Sometimes, if a 4-year-old is ready to enter kindergarten, they may enter that child, even if it is after the beginning of the school year.
Also, parents can be involved as much as they want to be in their children's education.
The school opened in Kittery in 1997, and moved to York in 2000. Herne started looking for schools for her son when she was still pregnant with him, and was disappointed with what she saw.
"Why should the philosophy go by the wayside," she said, and started a business plan for her own school. After she received financial backing, the school opened with one teacher and four students. It grew from there, and now the school has eight teachers, and 98 students.
She wanted to teach the students about the environment. Each member of the staff is required to have training in either wildlife management or environmental studies.
"We have a really high quality of educators," she said. Several of the teachers have bachelor's degrees, and the master teacher, Archer Israel, has a Ph.D.
Herne's commitment to the environment started at an early age.
Herne's father used to bring home animals all the time, such as snakes or frogs in a coffee cup. Today, the school has several animals, including snakes, frogs, turtles, and rabbits.
According to Herne, if a child is sad, they are sent to clean an animal, and then they suddenly become very happy. Also, with the animals, children learn about empathy and caring, Herne said.
On Sept. 1, 2002, the school was issued a license by the Maine Department of Education, recognizing the school as a private elementary school, which means it can use public school facilities.
If you are interested in enrolling your child into the Herne Schools of Discovery, they do have a screening process. "We look for people that have some sort of values, and that makes a huge difference," Herne said. She said a parent has to have a vision of commitment and future, and most be environmentally aware.
There are also scholarships available. Herne said the school is not like a "rich person's school, where only people who can afford get to come."
For information, call the school at 363-8631.

domain: news