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The objective of this study was to examine the expression of TLR by human primary uterine epithelial cells (UEC) and to determine whether exposure to the TLR agonist poly(I:C) would induce an antiviral response. The secretion of several cytokines and chemokines was examined as well as the mRNA expression of human β-defensin-1 and -2 (HBD1 and HBD2), IFN-β, and the IFN-β-stimulated genes myxovirus resistance gene 1 and 2′,5′ oligoadenylate synthetase. The expression of TLR1–9 by UEC was demonstrated by RT-PCR, with only TLR10 not expressed. Stimulation of UEC with the TLR3 agonist poly(I:C) induced the expression of the proinflammatory cytokines TNF-α, IL-6, GM-CSF, and G-CSF, as well as the chemokines CXCL8/IL-8, CCL2/MCP-1, and CCL4/MIP-1β. In addition, poly(I:C) exposure induced the mRNA expression of HBD1 and HBD2 by 6- and 4-fold, respectively. Furthermore, upon exposure to poly(I:C) UEC initiated a potent antiviral response resulting in the induction of IFN-β mRNA expression 70-fold and myxovirus resistance gene 1 and 2′,5′ oligoadenylate synthetase mRNA expression (107- and 96-fold), respectively. These results suggest that epithelial cells that line the uterine cavity are sensitive to viral infection and/or exposure to viral dsRNA released from killed epithelial cells. Not only do UEC release proinflammatory cytokines and chemokines that mediate the initiation of an inflammatory response and recruitment of immune cells to the site of infection, but they also express β-defensins, IFN-β, and IFN-β-stimulated genes that can have a direct inhibiting effect on viral replication.
Epithelial cells of the female reproductive tract (FRT)3 are the first line of defense against sexually transmitted diseases and invading pathogens. Understanding what role epithelial cells of the FRT play in immune surveillance and host defense is crucial for the development of effective mucosal vaccines as well as therapies for ongoing infections. Known to be an efficient physical barrier to infection, epithelial cells are in constant contact with the normal flora of the FRT and must discriminate between commensal organisms and pathogens (1). For the purpose of limiting the growth of commensal organisms on their external surface and defense of the underlying tissues from invading pathogens, epithelial cells have evolved innate immune antimicrobial functions as well as the ability to modulate the recruitment and activity of immune cells of both the innate and adaptive immune systems (2).
Important mediators of microbial recognition are germline-encoded receptors called TLR. TLR recognize conserved pathogen-associated molecular patterns (PAMP) synthesized by microorganisms but not by the host. Members of the TLR family, of which at least 11 TLR have been identified, recognize distinct PAMP produced by various bacterial, fungal, and viral pathogens. The recognition of bacterial PAMP, such as LPS, peptidoglycan, and flagellin, are mediated by TLR1, 2, 4, 5, and 6 (3, 4, 5, 6, 7, 8, 9). TLR7 and TLR8 recognize nucleotide derivatives, such as imiquimod, resiquimod, and loxoribine, and TLR9 binds unmethylated DNA found in bacteria (10, 11, 12, 13). In addition, recent reports have shown that TLR7 and TLR8 recognize both self and viral ssRNA (14, 15). TLR3 recognizes dsRNA, and more recently has been shown to be stimulated by cellular mRNA, potentially from cells killed by an invading pathogen (16, 17). The specific ligands for TLR10 and TLR11 have yet to be demonstrated (18, 19).
Epithelial cells throughout the body have been shown to express a wide range of TLR. Airway epithelial cells express TLR1–10, intestinal epithelial cells express TLR1–4, 6, and 9, and gastric epithelial cells express TLR2, 4, and 5 (20, 21, 22, 23, 24). Epithelial cells of the human FRT have also been shown to express a variety of TLR, including TLR1–3, 5, and 6 by vaginal and cervical epithelial cell lines and TLR1–3 and TLR6 by primary endocervical epithelial cells (25). We have previously shown that the uterine epithelial cell (UEC) line ECC-1 expressed TLR1–9 (26), but little work has been done to characterize the expression of TLR in human primary UEC.
Stimulation of human intestinal and pulmonary epithelial cells, as well as cell lines from the reproductive tract, with TLR agonists has been shown to induce proinflammatory cytokines and chemokines. TLR2 and TLR5 agonists induce the production of IL-6, CCL20/MIP-3α, CXCL8/IL-8, CXCL1/GROα, and CCL2/MCP-1 (24, 26, 27, 28, 29). TLR4 and TLR9 agonists induce epithelial cells to produce IL-6 and CXCL8/IL-8 (20, 30). Moreover, stimulation of epithelial cells has also been shown to induce the expression of small antimicrobial peptides called defensins (27, 29).
Defensins are small cysteine-rich, cationic peptides that are divided into three subfamilies: α-, β-, and θ-defensins. In humans, α-defensins are expressed in neutrophils, certain macrophage populations, and paneth cells of the small intestine and θ-defensin has been purified from the leukocytes and bone marrow of rhesus macaques (31, 32, 33, 34). In contrast, human β-defensins (HBD), of which four have been identified (HBD1–4), are largely expressed in various epithelial tissues, including skin, lung, and FRT (35, 36, 37, 38, 39, 40, 41, 42). HBD1 is generally thought to be constitutively expressed, while HBD2–4 have been shown to be highly inducible (35, 38, 39).
The roles that defensins play in defense of the host from pathogens are multifaceted, ranging from direct killing of invading microbes to linking innate and acquired immunities (31, 39, 41). Defensins are positively charged and interact with negatively charged components of microbial membranes and facilitate the permeabilization of Gram-positive and Gram-negative bacteria, fungi, and enveloped viruses (reviewed in Refs.38 and 42). Defensins are also crucial to the recruitment of immune cells to the sites of pathogenic challenge. HBD1–3 are chemotactic for immature dendritic cells (iDCs), CD4+CD45RO+ memory T lymphocytes, and mast cells, while HBD3 and HBD4 elicit monocyte chemotaxis (31, 35, 36, 43, 44). Although defensins have inhibitory effects on certain viruses, broad antiviral innate immunity is thought to be mediated by type I IFNs.
The IFN system is the first line of defense against viral pathogens and is crucial for limiting early replication and spread of viruses (45). Type I IFNs, known also as viral IFNs, exert their activity through the IFN-αβ receptor and include IFN-α, -β, -δ, -τ, -κ, and -ω. Production of type I IFNs are induced by dsRNA, which is synthesized by most viruses during their replication cycle (46), and by viral glycoproteins, including HIV gp120 (47, 48, 49, 50, 51). Recognition of dsRNA is mediated by TLR3, and expression of TLR3 is up-regulated following treatment of cells with IFN-αβ (52). Although all type I IFNs are important for an effective antiviral response, IFN-β is crucial to this process, because its absence results in the host being highly susceptible to viral infection (53).
Type I IFNs induce the synthesis of antiviral factors, such as 2′,5′-oligoadenylate synthetase (2′,5′-OAS) and myxovirus resistance gene A (MxA) (reviewed in Ref.54). Type I IFN induces the expression of the inactive form of 2′,5′-OAS, followed by activation of this enzyme in the presence of dsRNA. Once activated, 2′,5′-OAS polymerizes ATP into 2′,5′-linked oligoadenylates that are specific activators of a latent endoribonuclease, RNase L (55). RNase L degrades viral and cellular RNAs resulting in inhibition of protein synthesis (56). The 2′,5′-OAS/RNase L pathway has been shown to inhibit the replication of viruses belonging to different families, such as Picornaviridae (57, 58), Hepadnaviridae (59), Paramyxoviridae (60), Herpesviridae (61), and Retroviridae (HIV-1) (62, 63, 64). MxA belongs to the GTPase superfamily of dynamin-like GTPases (65, 66). MxA proteins associate with viral protein complexes and possess an intrinsic antiviral activity, although the mechanism by which MxA inhibits viral replication is poorly understood. MxA has been shown to be a crucial component in IFN-induced defense against several viral families, including Bunyaviridae (67, 68, 69, 70), Hepadnaviridae (71), Orthomyxoviridae (72, 73), Paramyxoviridae (74), Picornaviridae (75), Rhabdoviridae (73), Togaviridae (76), and possibly Retroviridae (77).
The goal of this study was to identify the TLR expressed by UEC and to determine whether exposure of epithelial cells to the TLR3 agonist poly(I:C) results in the secretion of proinflammatory cytokines and/or chemokines. Furthermore, the expression of antimicrobial defensins and antiviral genes was also examined following exposure of UEC to poly(I:C).
Materials and Methods
Source of uterine tissue
Uterine mucosal tissue was obtained immediately following surgery from premenopausal women who had undergone hysterectomies at Dartmouth-Hitchcock Medical Center (Lebanon, NH). Tissues used in this study were distal to the sites of pathology and were determined to be unaffected with disease upon inspection by a trained pathologist. Pathologists also determined the stage in the cycle of the premenopausal patients. Tissues were transported from Pathology on ice and procedures to prepare purified epithelial sheets began within 2 h of surgery. Approval to use tissues was previously obtained from the Committee for the Protection of Human Subjects (CPHS). Menstrual status had no influence on the experiments done in this study.
Isolation of UEC
Epithelial cells were isolated as previously described (78, 79). Briefly, tissues were minced under sterile conditions into 1- to 2-mm fragments and subjected to enzymatic digestion using a "PHC" enzyme mixture that contained final concentrations of 3.4 mg/ml pancreatin (Invitrogen Life Technologies), 0.1 mg/ml hyaluronidase (Worthington Biochemical), 1.6 mg/ml collagenase (Worthington Biochemical), and 2 mg/ml d-glucose, in 1× HBSS (Invitrogen Life Technologies) containing 50 U/ml penicillin and 50 mg/ml streptomycin. Enzymes were chosen to maximize digestion of the extracellular matrix while minimizing digestion of cell surface Ags. After incubating in PHC-HBSS for 1 h at 37°C, cells were dispersed through a 250-μm mesh screen, washed, resuspended in DMEM/F12 complete medium, and analyzed for cell number and viability. Complete medium was supplemented with 20 mM HEPES, 50 U/ml penicillin, 50 mg/ml streptomycin, 1 mg/ml fungizone, 2 mM l-glutamine (all from Invitrogen Life Technologies), and 10% defined FBS (HyClone) and did not contain phenol red.
Epithelial cell sheets were separated from stromal cells by filtration through a 20-μm nylon mesh filter (Small Parts). Epithelial sheets were retained on the 20-μm filter, while stromal cells passed through the filter. Epithelial sheets were recovered by washing and backwashing the filter with complete medium. Epithelial sheets were collected, centrifuged at 500 × g for 10 min, and resuspended in a small volume of complete medium.
Cell culture
To establish a cell culture system of polarized human UEC with both apical and basolateral compartments, the human UEC were cultured in Human Extracellular Matrix (Collaborative Biomedical Products) coated Falcon cell culture inserts in 24-well culture dishes designed for these cell inserts (Fisher Scientific). For these experiments, apical and basolateral compartments had 300 and 850 μl of complete medium, respectively. The medium was changed every 2 days. Following a 24-h incubation with various TLR agonists: ultra pure LPS from Salmonella minnesota (List Biological Laboratories) used at a final concentration of 1 μg/ml; Pam3cys-Ser-(Lys)4 (EMC Microcollections), 1 μg/ml; poly(I:C) (Invivogen), 25 μg/ml; Flagellin from Escherichia coli (Inotek Pharmaceuticals), 100 ng/ml; Zymosan from Saccharomyces cerevisiae (Invivogen), 10 μg/ml; Peptidoglycan from Staphylococcus aureus (Invivogen), 10 μg/ml: CpG oligonucleotide (Invivogen), 1 μM, UEC apical and basolateral supernatants were centrifuged for 5 min at 10,000 × g. For Ab-blocking experiments, UEC were pretreated with anti-TLR3 mAb (clone TLR3.7) (Cell Sciences) or an IgG1 isotype control (clone MOPC-21) (BD Pharmingen) at a final concentration of 20 μg/ml for 1 h at 37°C then stimulated with poly(I:C) for 24 h.
Measurement of transepithelial resistance (TER)
As an indicator of tight junction formation of epithelial cell monolayers, TER was periodically assessed using an EVOM electrode and Voltohmmeter (World Precision Instruments), as described previously (80).
RT-PCR analysis
Total RNA was isolated from cells using TRIzol Reagent according to the manufacturer's recommendations (Invitrogen Life Technologies) and purified with RNeasy columns (Qiagen). Two micrograms of total RNA were reverse-transcribed using the Superscript First-Strand Synthesis System for RT-PCR according to the manufacturer's recommendations (Invitrogen Life Technologies). PCR amplification was performed using Platinum PCR Supermix (Invitrogen Life Technologies) on the PTC-100 Thermal Cycler (MJ Research) for 35 cycles using the following cycling conditions: 94°C for 1 min, followed by 35 cycles of 94°C for 30 s, 55°C for 30 s, 72°C for 1 min, and then a final extension of 72°C for 2 min. Forward and reverse primer pairs for the TLR have been previously described (26). Forward and reverse primer pairs for HBD1 were HBD1 forward (5′-GTCAGCTCAGCCTCCAAAGG-3′) AND HBD1 reverse (5′CTTCTGCGTCATTTCTTCTG-3′); primer pairs for HBD2 were HBD2F2 (5′-CCTGATGCCTCTTCCAGGTG-3′) AND HBD2 reverse (5′-GAGGGAGCCCTTTCTGAATC-3′); primer pairs for HBD3 were HBD3 forward (5′-AGCCTAGCAGCTATGAGGATC-3′) AND HBD3 reverse (5′-CTTTCTTCGGCAGCATTTTC-3′). Lack of DNA contamination in the RNA preparations was verified by PCR amplification in the absence of reverse transcription.
Multiplex cytokine assays
Cytokines were measured using Bio-Plex human cytokine multiplex kits (Bio-Rad). Calibration curves from recombinant cytokine standards were prepared with serial dilutions in the same medium as the culture supernatants (RPMI 1640 medium containing 10% FBS). High and low reference points were included to determine cytokine recovery. Standards and reference points were measured in triplicate, each sample was measured once, and blank values were subtracted from all readings. All assays were conducted directly in a 96-well filtration plate (Millipore) at room temperature and protected from light. Briefly, wells were prewet with 100 μl of PBS containing 1% BSA, then beads (5000 beads per cytokine) together with either a standard, sample, reference point or blank in a final volume of 100 μl were incubated together at room temperature for 30 min with continuous shaking. Beads were washed three times with 100 μl of PBS containing 1% BSA and 0.05% Tween 20. A mixture of biotinylated Abs (50 μl/well) was added to beads for a further 30-min incubation with continuous shaking. Beads were washed three times, then streptavidin-PE was added for 10 min. Beads were again washed three times and resuspended in 125 μl of PBS containing 1% BSA and 0.05% Tween 20. The fluorescence intensity of the beads was measured using the Bio-Plex array reader. Bio-Plex Manager software with five-parametric-curve fitting (Bio-Rad technical note 2861 at <www.Bio-Rad.com>) was used for data analysis. The lower level of detection for IFN-γ was 37.5 pg/well; GM-CSF, IL-17, and CCL2/MCP-1 was 4.7 pg/well; IL-4, G-CSF, IL-6, IL-10, IL-1β, IL-12, and CCL4/MIP-1β was 2.3 pg/well.
Measurement of cytokine/chemokine and HBD2 secretion
Concentrations of CXCL8/IL-8, CCL2/MCP-1, CCL4/MIP-1β, TNF-α, IL-6, G-CSF, and GM-CSF in the apical and basolateral supernatants from human primary UEC were each determined with an ELISA test kit (R&D Systems) according to the manufacturer's protocol. Concentrations of HBD2 in the apical and basolateral supernatants were also determined by ELISA (Peprotech). The CXCL8/IL-8, CCL2/MCP-1, CCL4/MIP-1β, TNF-α, IL-6, G-CSF, GM-CSF, and HBD2 ELISAs had minimum detection levels of 2.3 pg/well. Standards for each ELISA were resuspended in cell culture medium. Calculations of cytokines, chemokines, and HBD2 were determined from a standard curve after OD measurements at 450 nm on an ELISA reader (Dynex).
TaqMan real-time RT-PCR
Real-time RT-PCR was done with a two-step protocol as described previously (81). Total RNA was isolated from cells using TRIzol Reagent according to the manufacturer's recommendations (Invitrogen Life Technologies) and purified with RNeasy columns (Qiagen). Coincident with RNA purification, was on-column DNase digestion using the RNase-Free DNase set (Qiagen). For each specimen, 600 ng of total RNA was reverse-transcribed using the iScript cDNA synthesis kit according to the manufacturer's recommendations (Bio-Rad) in a 20-ul volume. Relative expression levels of HBD1, HBD2, and HBD3 were measured using the 5′ fluorogenic nuclease assay in real-time quantitative PCR using TaqMan chemistry on the ABI 7700 Prism real-time PCR instrument (Applied Biosystems). The HBD1, HBD2, IFN-β, 2′,5′-OAS, MxA, and CD71 primer/MGB probe sets were obtained from Applied Biosystems assays-on-demand (ID nos. Hs00608345, Hs00823638, Hs00277188, Hs00242943, Hs00182073, and Hs99999911, respectively). Primers and probe used for detection of HBD3 are as previously described (82). HBD3 primers were used at 300 nM and probe at 50 nM. PCR was conducted using the following cycle parameters: 95°C, 12 min for 1 cycle (95°C, 20 s; 60°C, 1 min), for 40 cycles. Analysis was conducted using the sequence detection software supplied with the ABI 7700. The software calculates the threshold cycle (Ct) for each reaction and this was used to quantitate the amount of starting template in the reaction. The Ct values for each set of duplicate reactions were averaged for all subsequent calculations. A difference in Ct values (ΔCt) was calculated for each gene by taking the mean Ct of gene of interest and subtracting the mean Ct for CD71 for each cDNA sample. Assuming that each reaction functions at 100% PCR efficiency, a difference of one Ct represents a 2-fold difference. Relative expression levels were expressed as a fold-increase in mRNA expression and calculated using the formula 2−ΔΔCt.
The data are presented as the mean ± SE. A two-tailed paired t test or a one-way ANOVA with Bonferonni's posttest was performed using GraphPad InStat version 3.0a (GraphPad Software). A p value of <0.05 was taken as indicative of statistical significance.
Human UEC express TLR1–9
The expression of TLR1–10 UEC was examined using RT-PCR. UEC were isolated from hysterectomy tissue samples and grown to confluence on cell inserts coated with human extracellular matrix. The formation of tight junctions, as indicated by TER measurements of >500 ohm/well or greater (control wells had TER of 130–150 ohm/well), is an indication of UEC integrity and verification that a polarized monolayer has been formed. Once sufficient TER was achieved, total RNA was isolated and mRNA expression of TLR1–10 was examined. As seen in Fig. 1, UEC express mRNA for TLR1–9, while expression of TLR10 mRNA was not observed. The presence of TLR1–9 was observed in UEC preparations from eight different patients, and in all cases TLR10 mRNA was not observed.
The TLR3 agonist poly(I:C) induces the secretion of proinflammatory cytokines/chemokines by UEC
Having demonstrated that UEC express mRNA for TLR1–9, we examined whether stimulation of various TLR agonists would induce the secretion of cytokines and/or chemokines. UEC obtained from two patients (patients 2739 and 2749) were treated both apically and basolaterally with agonists to TLR2, TLR3, TLR4, TLR5, and TLR9 for a 24-h time-period, after which conditioned apical and basolateral medium were collected and screened for the expression of several cytokines and chemokines using the Luminex multiplex system. A greater amount of protein in the conditioned apical medium would signify preferential secretion into the lumen of the uterus, while a greater amount of protein in the conditioned basolateral medium would signify preferential secretion into the stromal tissues. Only the TLR3 agonist poly(I:C) stimulated UEC, while the other TLR agonists had no effect on the secretion of the cytokines and chemokines examined (data not shown). As seen in Table I, the levels of IL-6 and G-CSF secretion into the apical compartment exceeded the upper limit of sensitivity. Poly(I:C) induced the apical secretion of TNF-α, GM-CSF, CCL2/MCP-1, and CCL4/MIP-1β. Poly(I:C) induced the basolateral secretion of TNF-α, IL-6, GM-CSF, G-CSF, CCL2/MCP-1, and CCL4/MIP-1β (Table I). Poly(I:C) had no effect on the other cytokines examined, IFN-γ, IL-4, IL-1β, IL-10, IL-12, and IL-17, either apically or basolaterally (data not shown). These results demonstrated that UEC exposed to poly(I:C) induced the apical and/or basolateral secretion of four cytokines, TNF-α, IL-6, GM-CSF, and G-CSF and two chemokines CCL2/MCP-1 and CCL4/MIP-1β.
|IFN-γ .
|TNF-α .
|IL-1β .
|2739 Apical
|No PAMP
|2749 Apical
|No PAMP
|2739 Basolateral
|No PAMP
|2749 Basolateral
|No PAMP
|IFN-γ .
|TNF-α .
|IL-1β .
|2739 Apical
|No PAMP
|2749 Apical
|No PAMP
|2739 Basolateral
|No PAMP
|2749 Basolateral
|No PAMP
Conditioned media were collected following 24-h PAMP stimulation and were analyzed for cytokine production secretion using the Luminex multiplex system. Cytokines are expressed in total picograms.
Above upper limit of detection, IL-6 (1888 pg/well), G-CSF (920 pg/well).
Poly(I:C) induces the secretion of proinflammatory cytokines TNF-α, IL-6, G-CSF, and GM-CSF by UEC
Having used the Luminex multiplex system to screen for differentially secreted cytokines, we focused on TNF-α, IL-6, G-CSF, and GM-CSF as being induced by poly(I:C), and sought to confirm and extend these results by ELISA using UEC isolated from six patients. The baseline apical and basolateral secretion by UEC of TNF-α, IL-6, G-CSF, and GM-CSF are shown in Table II. The values are presented as the mean baseline secretion (nanograms per well) for the six patients examined as well as the range of secretion observed. TNF-α, IL-6, G-CSF, and GM-CSF were preferentially secreted into the apical compartment, with IL-6 secretion being ∼3- to 6-fold greater than TNF-α, G-CSF, and GM-CSF.
|Baseline Secretion (ng/well) .
|TNF-α .
|CCL4/MIP-1β .
|2.67 ± 0.27
|0.36 ± 0.15
|0.94 ± 0.24
|0.54 ± 0.14
|22.17 ± 3.98
|4.64 ± 1.40
|0.013 ± 0.003
|0.27 ± 0.12
|0.10 ± 0.03
|0.20 ± 0.05
|0.16 ± 0.08
|10.22 ± 1.57
|1.27 ± 0.38
|0.005 ± 0.003
|Baseline Secretion (ng/well) .
|TNF-α .
|CCL4/MIP-1β .
|2.67 ± 0.27
|0.36 ± 0.15
|0.94 ± 0.24
|0.54 ± 0.14
|22.17 ± 3.98
|4.64 ± 1.40
|0.013 ± 0.003
|0.27 ± 0.12
|0.10 ± 0.03
|0.20 ± 0.05
|0.16 ± 0.08
|10.22 ± 1.57
|1.27 ± 0.38
|0.005 ± 0.003
Values expressed as mean baseline secretion ± SE with range listed below each value in parentheses. UD, undetectable (number of patients = 6).
To more fully define the role of poly(I:C), UEC were treated for 24 h before analysis of apical and basolateral medium for the secretion of TNF-α, IL-6, G-CSF, GM-CSF by ELISA. As seen in Fig. 2, exposure of UEC to the TLR3 agonist poly(I:C) significantly induced the apical and basolateral secretion of TNF-α, IL-6, G-CSF, and GM-CSF. These results demonstrate that exposure of UEC to poly(I:C) induces the apical and basolateral secretion of proinflammatory cytokines TNF-α, IL-6, G-CSF, and GM-CSF without affecting the preferential directional cytokine release.
Poly(I:C) induces the secretion of chemokines CCL2/MCP-1, CCL4/MIP-1β, and CXCL8/IL-8 by UEC
The Luminex multiplex system also implicated the secretion of the chemokines CCL2/MCP-1 and CCL4/MIP-1β as being induced upon exposure of UEC to poly(I:C). Although not included in the Luminex multiplex screening, the secretion of CXCL8/IL-8 was included in these analyses because previous work done using the human UEC line ECC-1 demonstrated that CXCL8/IL-8 secretion was induced upon exposure of these cells to agonists TLR2 and TLR5 (26). Using identical supernatants that were examined for cytokine secretion, the apical and basolateral release of CCL2/MCP-1, CCL4/MIP-1β, and CXCL8/IL-8 was examined by ELISA. The baseline apical and basolateral secretion by UEC for CCL2/MCP-1, CCL4/MIP-1β, and CXCL8/IL-8 are shown in Table II. CCL2/MCP-1, CCL4/MIP-1β, and CXCL8/IL-8 were preferentially secreted into the apical compartment, with the secretion levels of CCL2/MCP-1 and CXCL8/IL-8 exceeding those of TNF-α, IL-6, G-CSF, and GM-CSF. For example, the apical secretion of CXCL8/IL-8 was >24 ng/well, which was nearly one to two logs greater than the secretion of TNF-α, IL-6, G-CSF, GM-CSF (Table II). As seen in Fig. 3, exposure of UEC to poly(I:C) significantly induced the apical and basolateral secretion of CCL2/MCP-1, CXCL8/IL-8, and CCL4/MIP-1β. These results demonstrate that exposure of UEC with the TLR3 agonist poly(I:C) induces the apical and/or basolateral secretion of the chemokines CCL2/MCP-1, CCL4/MIP-1β, and CXCL8/IL-8.
HBD1 and HBD2 mRNA expression is induced by UEC exposed to poly(I:C)
Because exposure of bronchial epithelial cells to poly(I:C) has previously been shown to induce β-defensin expression (49), we sought to determine whether exposure of UEC to poly(I:C) had any effect on the expression of HBD1, HBD2, and HBD3 (HBD1–3). Coincident with removal of apical and basolateral supernatants in previous experiments was the isolation of total RNA from the UEC. To determine whether UEC constitutively express HBD1–3, RT-PCR was performed. UEC were observed to express HBD1–3 mRNA (Fig. 4,A). Using real-time RT-PCR, we sought to determine whether poly(I:C) had an effect on the mRNA expression of HBD1–3. As shown in Fig. 4 B, exposure of UEC to poly(I:C) significantly induced the mRNA expression of HBD1 and HBD2 5.9- and 4.0-fold, respectively. To verify that protein secretion was also induced by poly(I:C), HBD2 secretion into the apical and basolateral supernatants from four patients was examined by ELISA. Stimulation of UEC with poly(I:C) resulted in a 45% increase in apical release of HBD2 protein (control = 42.2 ± 0.67 pg, poly(I:C) = 61.3 ± 3.8 pg; p < 0.05), but had no effect on HBD2 secretion into the basolateral compartment. Poly(I:C) had no effect on HBD3 (data not shown). Agonists to TLR2, TLR4, TLR5, and TLR9 had no effect on the expression of HBD mRNA.
Exposure of UEC to poly(I:C) induces mRNA expression of IFN-β and IFN-β-stimulated antiviral genes, MxA and 2′,5′-OAS
Because TLR3 binds and responds to poly(I:C), a viral dsRNA analog, we undertook to determine whether the mRNA expression of the antiviral IFN, IFN-β, was induced by poly(I:C) when added to UEC. In addition, the mRNA expression of the IFN-β-stimulated antiviral genes MxA and 2′,5′-OAS were examined. As determined by real-time RT-PCR, exposure of UEC to poly(I:C) induced IFN-β mRNA expression 70-fold and the mRNA expression of MxA and 2′,5′-OAS 107- and 96-fold, respectively (Fig. 5). These results demonstrate that poly(I:C) not only induces secretion of proinflammatory cytokines and chemokines and expression of β-defensins by UEC, but also induces the expression of IFN-β and the antiviral genes MxA and 2′,5′-OAS, which are important mediators of an antiviral response. Agonists to TLR2, TLR4, TLR5, and TLR9 had no effect on the expression of IFN-β, MxA, or 2′,5′-OAS mRNA.
Preincubation of UEC with anti-TLR3 mAb partially inhibits TNF-α and CCL2/MCP-1 secretion
To determine whether TLR3 expressed at the cell surface was involved in this response, UEC were preincubated with an anti-TLR3 mAb or isotype control. As shown in Fig. 6, incubation of UEC with an anti-TLR3 mAb for 1 h before the addition of poly(I:C) significantly inhibited the apical and basolateral secretion of TNF-α and CCL2/MCP-1 relative to that seen with isotype control. Inhibition of IL-6, G-CSF, GM-CSF, CCL4/MIP-1β, and CXCL8/IL-8 apical and basolateral secretion were also observed, albeit to varying degrees (data not shown). In contrast, preincubation of UEC with anti-TLR3 mAb, before poly(I:C) stimulation, had no effect on HBD1, HBD2, IFN-β, MxA, or 2′,5′-OAS mRNA expression (Fig. 6 C). These results demonstrate that blocking TLR3 expressed at the cell surface selectively inhibits the secretion of cytokines and chemokines in response to poly(I:C), but had no effect on the transcription of β-defensin, IFN-β, and IFN-β-stimulated gene mRNA.
This study demonstrates that UEC express TLR1–9 and that only the TLR3 agonist poly(I:C) stimulates UEC to secrete several cytokines and chemokines, particularly those factors involved in proinflammatory responses. Furthermore, we show that TLR3 stimulation of UEC induces the mRNA expression of the antimicrobial peptides HBD1 and HBD2. Finally, TLR3-mediated stimulation of UEC induced the mRNA expression of IFN-β and the IFN-β-stimulated antiviral genes MxA and 2′,5′-OAS. To our knowledge, this is the first demonstration of an antiviral response generated by primary human UEC.
Once thought to be a sterile environment, the uterus is now known to be periodically exposed to organisms colonizing the vagina (83). Furthermore, the demonstration that macrospheres deposited in the cervix move through the cervix and uterus into the Fallopian tubes within minutes of deposition is supportive evidence that bacteria and viruses in the vagina and cervix routinely reach the uterus (84). Considering the importance of the uterus with respect to pregnancy and fertility, expression of a large repertoire of TLR by UEC would allow for an enhanced sensitivity to a wider range of foreign microbes to rapidly and efficiently initiate innate and adaptive immune responses. Previous studies from our laboratory have shown that mRNA from a human UEC line express TLR1–9 and that TLR expression varies throughout tissues of the FRT (26, 85). Consistent with our previous findings, we demonstrate here that UEC also express TLR1–9. Neither ECC-1 cells nor UEC express TLR10. Others have recently reported mRNA expression of TLR1–6 and TLR9 by primary UEC and the absence of TLR10 expression (86). In contrast, we demonstrate that TLR7 and TLR8 are also expressed by UEC.
Poly(I:C) is a synthetic mimic of viral dsRNA that binds TLR3. Our findings demonstrate TLR3-mediated stimulation of UEC, via poly(I:C), results in enhanced secretion of the proinflammatory cytokines and chemokines TNF-α, IL-6, G-CSF, GM-CSF, CXCL8/IL-8, CCL2/MCP-1, and CCL4/MIP-1β. We previously reported that the adenocarcinoma-derived UEC line ECC-1 was not stimulated by poly(I:C) (26). This difference is likely due to greatly reduced TLR3 expression by ECC-1 cells compared with UEC (our unpublished observation). Secretion of the CXCL8/IL-8, CCL2/MCP-1, and CCL4/MIP-1β are known to have profound effects on the recruitment of immune cells, such as macrophages, monocytes, DC, neutrophils, and T lymphocytes (57, 87, 88, 89, 90). In concert with chemokine-mediated recruitment, TNF-α, IL-6, G-CSF, and GM-CSF induce B cell responses and enhance T cell help (91, 92, 93, 94, 95), stimulate the proliferation, differentiation, maturation, and antimicrobial functions of neutrophils (96, 97), as well as augment the functions of monocytes, macrophages, and DC (98). Therefore, in response to an invading viral pathogen, UEC have the capability of recruiting and activating a variety of immune cells that are known to be essential in preventing viral infection and restricting viral replication until an adaptive immune response is generated. Additionally, baseline secretion of these cytokines and chemokines, in the absence of poly(I:C) stimulation, suggests that UEC may play an important role in maintaining immune readiness/surveillance in the absence of pathogens. Previously, we and others have shown that CD45 leukocytes are present in the uterine endometrium throughout the menstrual cycle (99). Our findings in the present study suggest that baseline secretion levels of CXCL8/IL-8, CCL2/MCP-1, and IL-6 by UEC play an important role in leukocyte migration to the uterus to both detect as well as protect against potential pathogens.
In addition to the induction of proinflammatory cytokines and chemokines, UEC in response to poly(I:C) express 6- and 4-fold higher levels of HBD1 and HBD2, respectively. To the best of our knowledge, this study is the first demonstration that UEC expression of β-defensins increase in response to the TLR3 agonist poly(I:C). Our finding that HBD1 is induced by UEC following poly(I:C) treatment was unexpected, because HBD1 is thought to be constitutively expressed and unaffected by external stimuli (100). HBD1 and HBD2 are antimicrobial peptides that are effective against nonviral agents such as Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus as well as exhibit antiviral activity toward adenovirus, HSV, rhinovirus, and HIV (101, 102, 103, 104, 105). Both HBD1 and HBD2 facilitate the recruitment of immune cells, such as memory T lymphocytes, iDC and mast cells (44). Although HBD1 and HBD2 have been shown to be effective at inhibiting replication of HIV, HSV-1, and HSV-2, it is unclear whether they would have an inhibitory effect on other viral pathogens of the FRT, such as EBV, hepatitis B virus, hepatitis C virus, CMV, and human papillomavirus (106, 107, 108, 109, 110, 111, 112, 113). That HBD1 mRNA expression is induced by the TLR3 agonist poly(I:C) in UEC suggests that, in addition to its known antimicrobial function, HBD1 may have a more important role in antiviral innate immunity.
We and others have previously shown that secretions from throughout the FRT exhibit both bactericidal and viricidal activity (114, 115). Acting through secreted products including secretory leukocyte protease inhibitor, β-defensins, etc., epithelial cells of the uterus have the potential to interfere with HIV as well as other viruses that are both cell-free and cell-associated in the ejaculate (116, 117, 118, 119, 120). Other investigators have previously reported the antiviral action of IFNs (reviewed in Ref.54). Building on these important observations, that cells are capable of intracellular viral protection, we found that poly(I:C) induced UEC to express high levels of IFN-β and IFN-β-stimulated genes MxA and 2′,5′-OAS. It is known that TLR3-mediated immune responses, induced by dsRNA or viral infection, result in the rapid production of IFN-β and IFN-β-stimulated genes, such as MxA and 2′,5′-OAS (121, 122, 123). Using a pulmonary epithelial cell line, others have shown that the expression of TLR3 is induced by poly(I:C) (122). We have recently confirmed this finding with UEC exposed to poly(I:C) (our unpublished observation). The present study demonstrates that UEC express IFN-β in response to the viral analog poly(I:C). To our knowledge, these studies are the first demonstration that human UEC express IFN-β, as well as antiviral molecules MxA and 2′,5′-OAS, in response to the viral dsRNA analog poly(I:C), and that this response is mediated in a TLR3-dependent manner. Beyond the induction of IFN-β and the expression of antiviral molecules, such as MxA and 2′,5′-OAS, the induction of IFN-β by UEC may have important effects on leukocytes present at or recruited to the site of viral exposure/infection. For example, IFN-β has been shown to enhance cytotoxic activity and the production of IFN-γ by NK cells, stimulate the maturation of DCs and differentiation of monocytes into DCs, rescue memory T cells from apoptosis, enhance T lymphocyte proliferation and Th1 responses, as well as up-regulate iNOS in monocytes and macrophages (124, 125, 126, 127, 128, 129, 130). Our studies suggest that by secreting IFN-β in response to viral challenge, epithelial cells act as sentinels at multiple levels to prevent and/or control viral infection
To determine whether TLR3 expressed on the cell surface is responsible for binding poly(I:C), UEC were preincubated with anti-TLR3 mAb before stimulation with poly(I:C). We found that poly(I:C)-mediated secretion of cytokines and chemokines was partially inhibited by preincubation with an anti-TLR3 mAb. These findings suggest that TLR3 expressed on the cell surface participates in the recognition of dsRNA and triggers downstream signals leading to cytokine and chemokine production. Others have shown that TLR3 is found both on the cell surface as well as within some cells (124). Moreover, neutralization of cell surface TLR3 has been shown to partially inhibit (50%) the secretion of IFN-β (121). Our findings of 25–50% inhibition of TNF-α and MCP-1 secretion by epithelial cells in response to poly(I:C) (Fig. 6) are consistent with these observations. Unexpectedly, we found that preincubation of UEC with anti-TLR3 mAb had no effect on poly(I:C) induced expression of HBD1, HBD2, IFN-β, MxA, or 2′,5′-OAS when exposed to poly(I:C). The differences seen between cytokine and chemokine secretion vs mRNA expression of HBD1, HBD2, IFN-β, MxA, and 2′,5′-OAS may be due to the differential kinetics of protein synthesis and mRNA transcription over a 24-h time period. An alternative interpretation, suggested from our studies, is that in UEC stimulation through cell surface TLR3 is linked to a signaling pathway that is distinct from TLR3 expressed in subcellular compartments. Others have shown that in response to stimulation of cell surface TLR3, the MyD88-dependent signaling pathway, which is known to act through NF-κB (131), results in the production and secretion of cytokines and chemokines. In contrast, internalized dsRNA binding to TLR3 localized in endosomal compartments (124) may favor a MyD88-independent pathway, which through the activation of IRF-3 (132), has been shown to result in the expression of IFN-β and IFN-β-stimulated genes. Considering that TLR4 is also localized at the cell surface and intracellularly (133), determining whether stimulation of TLR3 and/or TLR4 at these different locales triggers different signaling pathways may further our understanding of how TLR function as part of innate immunity.
In conclusion, our studies suggest that UEC are poised to respond to viral infection at several levels. Stimulation of UEC with dsRNA, in a TLR3-dependent manner, induces the secretion of proinflammatory cytokines and chemokines that are known to facilitate the recruitment of immune cells to the site(s) of viral infection. Concurrently, TLR3 stimulation of UEC enhances the expression and possible secretion of the antimicrobial peptides HBD1 and HBD2 that would have inhibitory effects on viral entry into susceptible cells and therefore slow viral replication. At the intracellular level, IFN-β production by UEC might induce the expression of IFN-β-stimulated genes that have potent antiviral properties. Overall, UEC appear primed and ready to respond to viral exposure and/or infection of the uterus in a TLR3-mediated manner, which, in turn, leads to the stimulation of IFN-β and IFN-β-stimulated genes that are essential for inhibiting or slowing viral replication until an effective adaptive response can be mounted. Further studies are needed to extend our understanding of how UEC are involved in innate immunity, particularly antiviral responses, which are crucial for the development of mucosal vaccines targeting pathogens of the FRT, as well as therapeutics for uterine infections and treatments for infertility.
We thank Dr. Jacqueline Smith for advice and assistance on the Luminex multiplex system; Dr. David G. Ginzinger for advice with TaqMan real-time RT-PCR; Drs. Vincent Memoli and Jorge Gonzalez (Department of Pathology), Shannon Shutz, Peter Seery, Elizabeth Rizzo, and Richard Merrill for tissue preparation; Linda Hallock and Barbara Schaeffer for clinical support, and Drs. Kris Strohbehn, Leslie Demars, Paul Hanissian, Robert Porter, Benjamin Mahlab, Paul Manganiello, William Young, Roger Young, and Richard Meter (Department of Obstetrics and Gynecology) for tissue procurement.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
This work was supported by National Institutes of Health Grants AI51877 (to C.R.W.), Immunology Training Grant T32 AI07363-12, and Norris Cotton Cancer Center Grant CA23108.
Abbreviations used in this paper: FRT, female reproductive tract; PAMP, pathogen-associated molecular pattern; UEC, uterine epithelial cell; HBD, human β-defensin; iDC, immature dendritic cell; 2′,5′-OAS, 2′,5′-oligoadenylate synthetase; Mx, myxovirus resistance gene; TER, transepithelial resistance; Ct, threshold cycle.


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Target Hire Date: Immediately
Location: Atlanta, GA
Tigris Fulfillment Partners is a privately-held national distributor of kitchen and bath products, based in Atlanta, GA. Our customers are the largest big box retailers, leading e-tailers, and traditional kitchen and bath shops across the country.
We started this business in 2017 to address the opportunity in the marketplace to serve these customers in a better and faster way, and we built this company to be great. Our founders have built multiple companies and will not settle for anything but market leadership.
With the backing of a multi-billion-dollar international investor and a team mixed with industry leaders and fresh new minds, Tigris is set up for success and has already experienced tremendous growth. In our first two years, we installed a world-class warehouse management platform and now serve our customers with a fulfillment center network reaching 90%+ of the U.S. population in 1-2 days.
Tigris is built on a foundation of integrity and strong partnerships with our customers and vendors, and we are looking to hire good people who are great!
Our Financial Operations Analysts focus on key functional area intersections at Tigris. They excel with traditional analytics, quickly learn new technologies and are very comfortable with large data sets, and enjoy navigating cross-functional opportunities to improve business operations. Our analysts may be charged with improving current purchasing processes, finding greater profitability in our carrier relationships, reducing customer payment disputes, or automating current manual processes.
With deep exposure to both operations and finance, our analysts have career growth opportunities in nearly all functional areas at Tigris. Future roles may include Purchasing Manager, Finance Manager, or Operations Manager.
We expect our Financial Operations Analysts to be:
We expect our Financial Operations Analysts to possess:
Our commitment to you:
In addition to a competitive base salary, benefits of this role include:
Interested candidates should e-mail us at email@example.com
Tigris Fulfillment Partners is an Equal Opportunity/Affirmative Action Employer (EOE and M/F/Disability/Veterans.) We are a drug free workplace, with pre-employment drug screening required. Employment is contingent upon successful completion of a background investigation.

subset: commoncrawl


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Source: Red tide: China deploys new class of strategic missile submarines next year (July 2013)
China's navy is expected to begin the first sea patrols next year of a new class of strategic missile submarines, highlighting a new and growing missile threat to the U.S. homeland, according to U.S. defense officials.
"We are anticipating that combat patrols of submarines carrying the new JL-2 submarine-launched ballistic missile will begin next year," said one official familiar with recent intelligence assessments of the Chinese strategic submarine force.
China has deployed three nuclear-armed ballistic missile submarines to a naval base in the South China Sea, according to a recent photo of the vessels that appeared on the Internet.
"China's advance in submarine capabilities is significant," Locklear said. "They possess a large and increasingly capable submarine force. China continues the production of ballistic missile submarines. … This will give China its first credible sea-based nuclear deterrent, probably before the end of 2014."
originally posted by: BASSPLYR
So the chinese got themselves a early 1970 era tech submarine. Good job china ... But it's 2014 and you guys still got a long way to go with your sub tech. Meanwhile all three of those subs have probably been stalked and thoroughly analyzed by us naval assets since they've left port. Probably under hauled all of them.
The JL-2 has a minimum range of 2,000 km, a maximum range greater than 7,200 km, and carries a payload of 1,050 to 2,800 kg. This payload can be equipped with either a single 1 MT yield nuclear warhead or between 3 and 8 MIRV warheads. The MIRV warheads probably weigh between 250 and 300 kg and can be set to a nuclear yield of 20, 90 or 150 kT. The missile will likely be capable of being equipped with penetration aids and decoys to decrease the effectiveness of missile defense systems. The JL-2 uses an inertial guidance system with stellar updates and a Global Positioning Satellite (GPS) system. It may employ the Bei Dou navigational satellite system. An accuracy of 300 m CEP has been suggested, but other reports suggest 150 m CEP, which seems to better reflect current guidance technology. 2
originally posted by: Maxatoria
a reply to: BASSPLYR
The question always is, can you guarantee you can destroy those missiles before they hit their target? if not then they've got some nice tools at any bargaining session


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"I know almost nothing about her - so I measure the strength and fragility of my grandmother's life by the markers of our history that she outlived. Colonialism, independence, nationalism, civil war, peace and the illusion of progress…Her memory was long and in the end completely erased."
This text accompanies a black-and-white portrait of an elderly woman. The image and the caption were submitted by her grandson to an Instagram page— Brown History. In this tale, we do not know the names of the grandmother or the grandson. We know very little about this woman who lived in Jaffna, Sri Lanka, a few years ago.
The grandson, while talking about his grandmother says, he remembers "how at night her eyes shone like hessonite gems at arm's length of a kerosene lamp."
And so, we can summon the image of a young boy looking at his grandmother in a small town in Sri Lanka during the '80s, seeking comfort in her gaze amid a civil war.
View this post on Instagram
This is just one of the hundreds of posts available on Brown History—a project, initiated by Montreal-based electrical engineer, Ahsun Zafar. Brown History aims to let the 'Vanquished' retell South Asian history.
And if you're wondering how can an Instagram page embark on such a massive journey, all you have to do is scroll through the page.
Right from Muhammad Ali's visit to India, to someone recounting how their parents met in pre-partition Punjab, to the racism that one experienced as a first-generation immigrant in the US, Brown History addresses a wide range of issues.
The page documents history in a very unconventional manner— it relies on stories submitted through the internet. And by this, Ahsun aims to let the 'losers' narrate South Asian history.
"Winston Churchill once said, 'history is written by the victors but with Brown History, it's written by the people who lost. We have all grown up reading textbooks written by people who won the war, people who wrote stories which they wanted to narrate, to make them look good. And Brown History attempts to change that. It gives people a platform to submit their stories, in a way by which they are acknowledged," says Ahsun.
View this post on Instagram
But it's no small task to "retell" South Asian history, when the region has been wrought with war, violence, enmity and division. However, Ahsun feels that's precisely why his project is important. "These stories allow people to see each other as human beings and not as statistics. There's so much division in South Asia, which has arisen due to the fear of the other. We are all afraid of the unknown. And the only way to get rid of this fear, is to get to know the people, you are afraid of," he says.
The project materialised in 2017 with Ahsun wanting to learn more about his history.
"As I diaspora kid, living in Canada, I wanted to know more about my roots. Brown History is essentially a conversation I am having with myself, where everybody else is listening in as well," he says.
And when asked if his project is an example of the Empire fighting back, Ahsun says his page does that unintentionally and automatically.
"It wasn't really about fighting back. When you tell your story and your truth or what you think is the truth, you are automatically fighting a power that tells you the opposite story. So, it is just being you, in a world that's constantly telling you who you are and what you should do," Ahsun says.
Certain posts on the page, give the viewer a sense of déjà vu.
For instance, a post that has a picture published in The New Yorker in 1987 shows a group of people holding placards that read "Hudson Against Racism". The caption further elaborates on the anti-racism movement prevalent in New Jersey at the time, something that emerged as a response to hate crimes against South Asians. The post makes one immediately draw parallels between the anti-racism movement then and the Black Lives Matter movement, now.
View this post on Instagram
When asked about this, Ahsun says, "History does repeat itself and if it does not repeat it definitely rhymes. There's always some new power that comes to be and it does whatever it can to maintain that power. Those who are not aware of the past fall into a trap and they just repeat the cycle. But if everybody understood the past and how communities that are in power oppress minorities, they would know better and that's why history is important."
But it is not easy to always demarcate documentation and activism and in fact many would say it's wrong to mix the two. And with a project like Brown History, the lines blur. However, Ahsun believes there's nothing wrong in it. "I try to stay neutral but if I am quiet and silent to injustice then I have automatically chosen the oppressor's side. And I cannot do that. I want no part in someone's oppression," he says.
But neutrality brings with it another set of problems, that of representation and portrayal. And when working with a group as diverse and conflicted as South Asians, criticism and allegations of bias are sure to rear their ugly head. And Ahsun admits that it's becoming more difficult, with every passing day.
"No matter what, you are going to piss off people. If I post something about Hindus, then Muslims attack me and vice versa. If I post about the treatment of Shias in Pakistan, then the rest of Pakistan will hate on me and say that I am making Pakistan look bad. But if you are a Shia Or Ahmadi Muslim, then the Pakistan that you live in, is very different compared to the Pakistan that an upper-class, Sunni Muslim lives in. But I Don't let the negativity get to me, I accept stories from everyone and do not discriminate," says Ahsun.
Further elaborating on this, he says that while history is complicated, each person/group is probably a victim in its own way and we all should "have the right to have our pain acknowledged".
Brown History began as a personal project and within three years it has gained more than 500K followers. And Ahsun has been overwhelmed with all the love and support he has received during this time.
"For every person, who abuses me, there are 10 others extending encouragement and support and that's what keeps me going," he says.
And now, the Montreal-based electrical engineer hopes to compile all that he has documented in the form of a book in the near future.


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VA Secretary Robert Wilkie told senators on Tuesday that the department is not recommending an appeal to the Procopio vs. Wilkie decision, a Jan. 29 court ruling that restored benefits to veterans who were exposed to Agent Orange during their service off the coast of Vietnam.
Until Tuesday's Senate Veterans' Affairs Committee hearing, VA officials were silent about whether they intended to contest the decision by appealing to the Supreme Court before the window for appeal closed at the end of April. Prior to the announcement, MOAA and other veteran advocacy groups wrote to President Donald Trump requesting that he tell the Department of Justice not to appeal the ruling.
[CALL TO ACTION: Ask Your Legislators to Support Blue Water Navy Veterans]
Estimates for the number of Blue Water veterans vary, but the Congressional Budget Office estimated in December that extending benefits to those veterans would increase direct spending by $1.2 billion between 2019 and 2028, although retroactive payments, compensation for survivors, and other associated costs could add to that figure.
If the VA follows through on Wilkie's statement, the administration will be responsible for that bill in coming years, requiring action from Congress to appropriate the funding to cover the influx of new disability claims.
The Blue Water Navy Vietnam Veterans Act, H.R. 299, is another way Congress could choose to address the cost of benefits for Blue Water Navy veterans. The bill includes adjustments to the VA home loan program, increasing the limit on the amount of loans eligible for the program and temporarily extending funding fees for veterans without disabilities.
MOAA remains active and engaged on the needs of Blue Water Navy veterans, and we encourage Congress to provide the VA with all the necessary resources to support implementation of these benefits as soon as possible.


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New Jersey, United States,- Market Research Intellect recently added the Beds on Casters Market research report which provides an in-depth scenario analysis of the market size, share, demand, growth, trends, and forecast for the market from 2020 to 2027. The report covers the impact analysis of the COVID-19 pandemic. The COVID-19 pandemic has affected export-import, demand, and industry trends and is expected to have an economic impact on the market. The report provides a comprehensive analysis of the impact of the pandemic on the entire industry and provides an overview of a post-COVID-19 market scenario.
The report majorly mentions definitions, classifications, applications, and market overview of the Beds on Casters industry. It also covers product portfolios, manufacturing processes, cost analysis, structures, and industry gross margin. It also provides a comprehensive analysis of major competitors, their regional breakdown, and market size.
The report covers extensive analysis of the key market players in the market, along with their business overview, expansion plans, and strategies. The key players studied in the report include:
read more: bulletinline.com


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[A version of this story ran in Pinoy Newsmagazine. Read that version here.]
Jessica O'Brien prepared herself for the worst.
She didn't plan a party for March 20, 2012, because she wasn't sure if she was going to win the Democratic Cook County Circuit Court primary election for the vacancy of Henry Simmons Jr.
"I didn't have any expectations,†said O'Brien, who admits that she's so competitive that family games of volleyball get heated. "I ran as if I was losing every single day.â€
At 7:30 P.M., a half hour after the polling places closed, she received her first congratulatory phone call from Justice Nathaniel House. She was so far ahead in the polls House believed no one would catch her. It wasn't real for O'Brien yet.
Her attitude was, "I'm not going to believe you until the fat lady sings,†she said.
Even after all of the votes were tallied the next morning, O'Brien refused to believe she had received the Democratic Party nomination. She couldn't be 100 percent sure until she received a letter from the political party in the mail 49 days later.
It was finally real for her. "Today, to me, it's official,†O'Brien said the day she received the letter. "It's still not sinking in.â€
This story is part of a unique collaboration of twelve Chicago area ethnic news media examining how different immigrant communities in and around Chicago are approaching the 2012 election campaigns. The stories were released in June 2012 by Africa Today, Al Moustaqbal – Future newspaper (Arab), Bulgaria Weekly, Draugas – The Lituanian World-wide Daily, Extra (Hispanic), India Tribune, InformacjeUSA.com (Polish), Korea Daily News, Pinoy Newsmagazine (Filipino), Reflejos (Hispanic), Reklama (Russian), and  Urdu Times (Pakistani). Click here to access the other stories: Indian Americans shift party allegiance and take their money with them, An election dilemma for Poles: Vote religion or immigration, Russian immigrants trending Republican, Russian Jews moreso, Bulgarian immigrants may abandon Obama, but voting participation unclear, Pakistani Americans mobilize for the elections…back in Pakistan, Memories of corruption and the Machine stunt Latino vote, Dual citizenship for Lithuanians not for everyone, For Arabs, there's more to American citizenship than voting, PERC leads Korean immigrant community into the political process, and African activism coming off the sidelines.
For O'Brien to win her party's nomination she overcame more than opposing candidates Michael Forti and James Wright. Law is the Filipino immigrant's second career, something that had always interested her, but a field she wasn't sure she had the foundation to pursue.
Historically Filipino-Americans are less politically active than other immigrant groups in Chicago. Experts have pointed to corrupt politics in the Philippines and a colonial mentality that remains in the psychological mindset of Filipino immigrants when they arrive in the United States. Filipino Americans who choose to run for public office tend to be second-generation immigrants.
O'Brien is an anomaly not only in the Filipino community, but in the Asian-American community as a whole. According to the United States Census Bureau, Asian-Americans represented 6.2 percent of Cook County's population in 2010. Of the 400 Circuit Court Judges only nine (or 2.25 percent of the circuit court) are of Asian-American descent.
The lack of Asian-American representation is likely why O'Brien was advised by members of the Democratic Party to leave her photo off all her campaign material. Her ethnic background could potentially cancel out any bump she'd get for being a female candidate with an Irish last name, according to a party official O'Brien chose not to name. However, O'Brien decided that a smile would help gain voters trust and ignored the advice.
"I want people to know I'm Filipino but really it shouldn't even be a factor,†she said. "I mean look at my credentials.â€
She wasn't talking about marrying Brendan O'Brien when she was involved in the hospitality business – long before she knew she'd be running for a spot on the Cook County Circuit Court, but rather her awards from the Asian American Bar Association and Women's Bar Association of Illinois.
"Race is secondary to me. Qualifications are always first,†she said. "It's good that minorities are involved, but they have to be qualified.â€
Other Filipino politicians in the Chicago area – such as Florencio Villegas, a Bloomingdale Township committeeman and Jerry Clarito, Vice President of the Board of Directors of the Skokie Park District – believe the low number of Filipino representatives in the Chicago area is due in large part to lack of political participation from the Filipino community as a whole.
"You've seen [other immigrant] groups rally,†said Villegas. "Filipinos have never been that way.â€
Villegas and Clarito point to two main causes of political apathy in their community. First, politics in the Philippines is associated with buying votes and violent elections. It's dangerous to be politically active there.
"Politics in the Philippines is not encouraging, to say a mild word,†said Clarito, who as a dual citizen has been politically active in both the Philippines and the United States. He opposed the U.S. support of Filipino dictator Ferdinand Marcos and met with U.S. congressmen to get the Filipino Veterans Equity Compensation Act, which gave financial compensation to Filipinos who were a part of the Bataan death march, passed.
Second, a colonial mentality is often mentioned as a contributing factor to low political participation. The Philippines was a colony of Spain for nearly 350 years from 1521-1898 and a U.S. colony from 1898-1946.
Kevin Nadal, a psychology professor at the John Jay College of Criminal Justice in New York City, has studied this colonial mentality extensively. In 2009 he published Filipino American Psychology: A Handbook of Theory, Research and Clinical Practice. Nadal believes that the effects of being a colonized people for such an extended period of time has made most Filipino immigrants feel inferior to U.S. citizens born in the United States.
"To be American is viewed as far more superior,†Nadal explained. "But [Filipino immigrants] don't fully assimilate. Instead, they don't quite feel American enough.â€
His 308-page book is in Villegas' living room. The 79 year old believes the best way to address both causes of Filipino voter apathy is to teach immigrants Filipino history without a U.S. slant to allow future generations to become more politically active.
"I'm almost embarrassed to address the issues because Filipinos, my generation anyway, aren't going to be involved,†said Villegas.
The Alliance of Filipinos for Immigrant Rights and Empowerment (AFIRE) is also dedicated to educating Filipinos about their colonial history and the importance of political participation.
The organization is part of the New Americans Initiative, a statewide program that focuses on easing the way for legal residents to become citizens when they are eligible. Ninety-two percent of Filipinos who immigrated to the United States from 1970-1979 became citizens, according to AFIRE. However every decade since, the number has declined. Only 12 percent of Filipinos who came to America from 2000-2009 are U.S. citizens.
AFIRE president Juanita Salvador-Burris says that the organization is already seeing improvement on these numbers because of their work with the New Americans Initiative.
The aim is to get more Filipinos registered to vote, and create a more politically educated base of Filipinos, which can influence lawmakers – something that the Filipino community has not done historically in the United States.
"A common expression among politicians is, 'you Filipinos are not getting your share,' which is so true.'' said Villegas. "We don't know how to pursue that share because we don't know how to play the game of politics. We have made no effort to understand it.â€
That doesn't apply to Jessica O'Brien. After winning the Democratic primary she is now basically a shoe-in for the Cook County Circuit Court.


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Dear Chamber Members and Partners:
Last Friday, Governor Greg Abbott announced the creation of the Strike Force to Open Texas, comprised of nationally recognized medical experts along with business and community leaders from throughout Texas, including substantial San Antonians, who will advise the Governor on how to safely and strategically reopen Texas and restart the state's economy. To assist the Strike Force, the Governor's office is requesting information on the return-to-work plans of Texas businesses.
Please take a few minutes to complete one of two available questionnaires before noon on Thursday, April 23, 2020, so that we can send your input to the Governor's office before the end of the day Thursday.
If your business is essential as deemed by the Department of Homeland Security/CISA guidelines, please complete this survey.
If your business is non-essential as deemed by the Department of Homeland Security/CISA guidelines, please complete this survey.
Please note that you do not have to answer every question, and your survey responses may be subject to public disclosure under the open records law.
We urge you to complete the survey to ensure the voice of the San Antonio business community is heard and considered in the plans to reopen the Texas economy, and we look forward to working with members of the Governor's Strike Force to Open Texas through the recovery period.
In the meantime, please continue to stay safe and healthy!
Richard Perez
President and CEO
San Antonio Chamber of Commerce


text: A NHS message is seen on a screen at a coronavirus disease (Covid-19) pop-up vaccination centre at Wembley Stadium in London, Britain, December 19, 2021. \u00a9 REUTERS\/Peter Nicholls Anti-vaxxers have been booking up Covid-19 vaccine appointments and then not turning up to get jabbed, according to the Telegraph, as Britain rushes to vaccinate millions against Omicron.Appointments at mass vaccination centers, including Wembley Stadium which is being used to roll out thousands of shots before Christmas, are being block-booked by people who have no intention of getting inoculated, according to the Telegraph on Sunday. \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n Read more\n Kremlin lashes out at 'dangerous fools'\n \n \n\nThe paper claimed that Wembley Stadium, where the anti-vaxxers had reportedly been employing their tactic, was quiet on Sunday morning. Staff told the Telegraph that patients had not arrived for their bookings and the center was "really quiet" by mid-morning.However, a spokesman for North West London Integrated Care System said that the center would still make a big contribution to the UK's inoculation effort. "Any suspected disruption has been offset by the number of walk-ins, including thousands of local residents, many of whom have made a big step in coming forwards to start their vaccination journey, as well as very high numbers of boosters," the spokesperson stated.The site was hoping to inoculate 10,000 people on Sunday, but only 2,500 had been administered by mid-afternoon, according to the Telegraph. It is understood that anti-vaxxers have employed similar tactics at other vaccination sites around the country.The UK has administered more than 28 million boosters to fight back against the spread of the Omicron variant during the festive season. More than 900,000 shots were administered on Saturday alone. Health Minister Sajid Javid has said that nine of out 10 patients needing the most care against Covid-19 were unvaccinated. "And I just cannot emphasise enough the impact that they are having on the rest of society," he stated.\r\nSource


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Every six months we like to see you for a checkup and cleanings to keep your smile strong and healthy. However, in between these visits you need to care for your smile from home to avoid issues like tooth decay and gum disease. In today's blog, your Celina, TX, dentist is talking about the importance of routine brushing and flossing.
The Dangers of Plaque and Tartar
Over time, a combination of foods and drinks high in sugar and starch, and poor oral hygiene habits, could enable plaque buildup to accumulate and coat your teeth. This bacterial byproduct is difficult to remove with brushing and flossing alone, and could eventually harden into tartar. The buildup not only weakens the enamel to cause tooth decay, but can inflame gum tissues to cause gingivitis and gum disease. The buildup is also linked to teeth stains and bad breath. To control the buildup, you need to take good care of your smile in between your six-month checkup and cleaning visits.
Brushing and Flossing Your Teeth
Daily brushing and flossing help control how much plaque can accumulate. These steps remove food particles to deprive bacteria of a meal. You should brush when you wake up for about two minutes, and do the same before going to bed that night. Each time, be sure you use a fluoride toothpaste, as this strengthens the outer enamel. You also need to floss every evening, as this removes what a toothbrush alone cannot. Move the tape up and down each side of every tooth, running up and down the entire length and moving below the gum line in a "c" motion. Our team would be happy to discuss tips for better homecare!
Eating a Healthier Diet
Remember, sugar and starch could feed bacteria and make buildup more severe. We suggest eating fewer sugary items, and instead adding more fresh fruits and vegetables, calcium-rich cheeses, and whole grain items to your diet. Drink more water, which helps cleanse your smile and contains no sugar or acids. In fact, your tap water even contains fluoride to protect your smile! Cut back on beverages like soda energy drinks, or sports drinks, all of which are linked to a higher cavity risk.
Common Signs of Trouble
In between your visits, be sure to let us know if you encounter tooth sensitivity, toothaches, or sore, bleeding gums. These are often indicators of issues like tooth decay and gum disease, and should be addressed right away. If you have any questions, give our team a call!
Talk To Your Celina, TX Dentist's Office About Homecare
Our team is ready to work with you and help you enjoy optimal oral health. To find out more about preventing oral health troubles, contact your Celina, TX, dentist, Dr. Ganjoor, by calling (214) 851-0130.


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Men's Lacrosse vs. St. Thomas Aquinas
Mar 9, 2020
A six-goal third quarter expanded the Wilmington University men's lacrosse team's lead as it went on to a 14-9 nonconference victory over St. Thomas Aquinas on Saturday at the WU Athletics Complex to remain undefeated. Freshman Logan Stinson scored six points, putting gin four goals while adding two assists. Giovanni Marino scored two goals while adding an assist, and Jacob Herman scored once and added a team high three assists. Jake Mollohan led the defense with three caused turnovers while Colyn Lyons and Jacob Herman had two each. Noah Given made 10 saves in net for the Wildcats.


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Epilepsy Foundation helped our daughter
EDITOR: When our daughter was diagnosed with epilepsy at the beginning of eighth grade, our family was caught unprepared. Years later, we still struggle with the devastating affect epilepsy has had on her life, but the Epilepsy Foundation Heart of Wisconsin has been an invaluable resource to us.
Our first call to the Epilepsy Foundation was literally a cry for help. We wondered whether she would lead a normal life; we hoped to find other parents who were in a similar situation; and we needed to learn more about medicines and treatment. The director was calm, soft-spoken and reassuring. We realized we were not alone. There was someone who would listen, someone who would understand and someone who could provide us with resources.
Since that time, in addition to helping us as caregivers, the Epilepsy Foundation has given our daughter a place to belong. She attends support group, stays overnight at the Adult Epilepsy Camp in Rosholt, participates in art therapy opportunities and other clinics, advocates for legislation to help those with disabilities, and walks in the annual Stroll for Epilepsy. She feels accepted and safe with her friends and the directors she has met through the Epilepsy Foundation. It has helped alleviate some of her loneliness, and it has helped her through the frustrations of this disease.
We all know that there is still much more to learn about epilepsy and there are more people to reach. Become an advocate. Although many have good seizure control, too many people do not. They are often isolated and alone. The Epilepsy Foundation Heart of Wisconsin promotes the cause.
For the good of all, please consider attending support group on the second Monday of each month, the summer picnic on Monday, July 13 at Bluegill Bay County Park, the family-friendly Stroll for Epilepsy on Aug. 22 in Marathon Park.
Your presence can make a difference. Call 715-341-5811 or email firstname.lastname@example.com for more information.
Barb Mullen,


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Sinopsis :
Emma Woodhouse, handsome, clever, and rich
Emma is also overly confident in her abilities as a matchmaker for the residents of Highbury in Jane Austen's widely beloved classic novel. Although convinced that she herself will never marry, Emma makes finding the perfect partner for her new friend, Harriet Smith, her most pressing project. It is a well-meaning endeavor that leads both women intoa complex tangle of intrigues, misunderstandings,and conflicts of affection, especially after several new admirers come to the fore whose sights are maddeningly set on the wrong woman. Matters of the heart are never as simple as they seem.
Jane Austen's charming novel of love, friendship, and social grace is a timeless classic as fresh, funny, and poignant as it was when first written.
Penulis : Jane Austen
Assalamualaikum Dengan toko buku zanafa saya ingi bertanya stok....
Jl. Prambanan No.877, Ruko ABCDEFG, Banyuwangi - Jawa Timur, Indonesia


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These vintage diamond sapphire dangling earrings are adorned with a combined weight of 2.95cts of brilliant round diamonds and multicolor sapphires. Marquise-shaped green sapphires, and pink and yellow round sapphires combine in these 14K white gold dangling earrings to create a refreshing, spring-time appearance. A special Dacarli design, they are sophisticated, chic and unique and are sure to provoke admiration–as well as conversation.
The characteristics of these earrings are milgrain, antique engraving and scrollwork. Dacarli has been manufacturing diamond jewelry for three generations, since 1939. When we make our earrings we are careful to produce pieces with a heavy gold weight, which ensures that they will give many years of trouble-free wear.
At Dacarli, emphasis is placed on design, quality, and Old World European craftmanship. Our earrings are manufactured in our Florida factory using the finest gold and G-H color, SI clarity conflict-free diamonds. We use only sapphires and other color gems of the finest color and cut.


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An article in the Business Times' Editorial & Opinion column of 25 Apr 2012 talked about developing global leaders with an Asian focus. I thought it was well-written and it discussed issues facing many leaders today.
What is an "Asian" leader? What is a "Western" leader? Is the "Asian" leader more "collaborative" and are "Western" leaders more "assertive"? I mentioned in a previous post that we have a tendency to dissect and draw lines rather than synergize. We seem to prefer the "OR" instead of the "AND". Think about it – we tend to look for "Specialists", do we not? Nothing wrong with "Specialists", just make sure that their foundations are large and stable enough to support whatever it is they are "Specializing" in. That's the point. A leader is a leader. A leader needs to inspire people to internalize truth and work towards an ennobling and shared goal. Regardless of whether he or she comes from the East or the West, a leader must have good character qualities. Period. A good leader serves those he leads. In government, why do you think leaders are called "Ministers"? A leader sees and knows what is good for his people. This is not necessarily what they WANT, but what they NEED. He has the guts to make tough calls when necessary. He is not afraid of being voted out of office. At the same time, he sets great store by the development of other leaders and of his people in general. He lives by the adage and philosophy of making his people "Greater Than Yourself". He sees what is for the greater good and steadfastly steers his people there, absorbing outcries and rants against him. He also has the foresight, boldness and willpower to both neutralize those that would manipulate people for their own ends and to invest the enormous resources to develop his people to be their best. When people are maximized, crime rates drop. Demand for addictive and destructive pursuits like alcohol, drugs, tobacco and moral licentiousness goes down. Productivity goes up, and people face the future confident in their shared ability to forge it instead of being victims to it. Where are such leaders today? I don't care if they are "Western" or "Asian" or come from Timbuktu. I care very much, however, that they be Genuine Leaders. Do you?


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It's been far too long since we've heard from our buddies, Maddie Wahl and Erin Kelly. But, they have pretty good excuses as Erin's been a writing and acting machine and Maddie is a freshman in college. And, as you can see, she's studying art. This is beautiful work that calls to mind the elegant simplicity of Mark Buckingham's style.
Welcome back, ladies!


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Christmas is the only festival which is celebrated all over the world in each country to celebrate the birth of Jesus Christ. Each year this day of Christmas is marked with great enthusiasm and good spirit by people of all religions and regions. Even in countries where Christianity is not a majority's religion has Christmas trees and various decorations all over the streets marking their love for this festival. It is interesting to how different countries celebrate Christmas and the traditions they follow. So here are the ways in which these 10 countries from different corners of the world celebrate Christmas !!
1.AUSTRALIA: Merry Christmas Mate!
Christmas in the land of Aussies is celebrated in the summer time. The 'White Christmases' is unheard on this land. It is a tradition, in the weeks up to Christmas to join the picnics organized by various churches and sing Christmas carols on the beach and big holiday pageants are broadcast.As it is the summer time, it is traditional to enjoy sizzling barbecues on this big day and Father Christmas swaps his reindeer for 'six white boomers' or kangaroos.
2. Brazil: Feliz Natal!
In the high temperatures, a flock of tourists and citizens rush to the beaches during these summer holidays and the 'sandmen' are replaced by 'snowmen' just like this Santa on the beach in the Rio.In Brazil, kids expect "Papai Noel" (the Father Christmas )to deliver the presents. They celebrate the legend that animals gain the power of speech on the Christmas night.
3. India: Subah Krisamas!
Due to lack of fir and pine trees on this sub-continent, banana and mango trees are often used as substitutes for Christmas trees and mango leaves are used to decorate and brighten up the homes.Many of the traditions here are influenced by the western cultures like the Christmas trees and plenty of holiday sweets, including fruit cake, coconut filled pastries called 'Nuereos'.And people go to churches for midnight masses to mark this big day.
4. France: Joyeux Noel!
In many regions of France, Christmas celebrations start with the St. Nicholas day on 6th of December. On the Christmas eve, children keep their polished shoes in front of the chimney and hope that "Papa Noel" will fill their shoes with sweets. The 25th of December is a public holiday and families get together for the big feast with good meat and the best wine.The decorations in the house are minimal but now the British traditions are being taken up and the town centers are decorated with the Christmas trees and decorations.
5.Iceland: Gledileg Jol!
This place has got a very interesting tradition.There are thirteen Santas in the Iceland. These Yule lads, more troll like than our traditional Father Christmas, who traditionally come down from the mountain one by one on these days of the festive period and leaving presents or rotten potatoes depending upon the child's behaviour.In Iceland, children put their shoes on the windowsill, so that the Santa Claus can fill their shoes with the goodies. Families get together here and visit churches for midnight mass.
6.Italy: Buon Natale!
In Italy, 'Babbo Natale' that's how the Father Christmas is called in Italian, hands out the presents to the children on the Christmas day.But still in many families, gifts are exchanged only on 6th January that is the day of Epiphany.
7. Netherlands: Zalig Kerstmis!
In Netherlands, Santa Claus is called Sinterklaas. He doesn't live at the north pole but in the sunny Spain and arrives by a steamboat with a helper called Black Peter rather than an elf.
8. USA: Merry Xmas!
In USA Christmas is celebrated on 25th and 26th of December. On the Christmas day, families get together for the feast after exchanging the gifts.On 26th people usually visit friends and families and exchange gifts. It is customary to send and give Christmas cards, on which some write 'Merry Xmas', where Xmas stands for Christmas and X signifies 'Cross'.
9. Germany: Frohe Weihnachten!
The Christmas season here starts with vibrant street markets, filled with lights, ornaments and other decorative items. The Christmas trees are usually put up in the homes on the Christmas Eve and festive celebrations begin from the Christmas eve and the market closes down after the lunchtime on 24th. People celebrate this day with vegetables, potatoes and a German cake.
10. Indonesia: Selamat Natal!
Here Christmas is locally known as 'Natal'. Churches and shopping malls are decked with lights and decorations. Here there is the tradition of going to masses twice, one at midnight and the other during the day is popular.Christmas celebrations bring back the life to the villages with colourful clothing and beautiful dancing.
Merry Christmas !!! HO! HO!HO!!!


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City University is located in Bellevue, Washington and is a private college. City University is a four year college and offers Associate's Degrees, Bachelor's Degrees, Master's Degrees, and a number of different programs and courses.
City University is in a relatively urban area (in or near a city), which may be something you prefer if you like a city lifestyle as a student.
City University has a rolling admissions policy, which means that you can apply throughout the year for the upcoming semester.
City University is relatively smaller in size with an enrollment of only 4,509 students.
City University offers military credit for military courses that have been completed successfully by students.
If you have taken some advanced placement courses with an applicable test, or obtained credit from an other college, you may be eligible to transfer that credit to City University.


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So much of this feature is dedicated today to a decade none of its editors saw — the 1960s. At the head, we have a Netflix tribute to one of the most influential comedy cavalcades of all time. HBO & ABC also offer gripping documentaries with a thematic thread of our then and still now racial divide. We also recommend two young shows featuring mostly actors who also weren't alive to see the sixties. Welp, time to, um, sock it to ya!
Still Laugh-In: The Stars Celebrate [Netflix]
If not one of the best shows in TV history, Rowan & Martin's Laugh-In is certainly one of its most important as, along with the Smothers Brothers Comedy Hour, it marks a solid bridge from old-school variety shows and half-hour comedies to the success of sketch shows in the SNL vein — and it had Goldie Hawn, Lily Tomlin, Ruth Buzzi, Arte Johnson, etc. Per the banner pic above, at least Tomlin and Buzzi will be there, along with admirers from Billy Crystal to Snoop Dogg. Although, can I get mad at a show from before I was born for normalizing a corrupt President who paved his way for our current debacle or do I admire the fact they were able to get Richard Milhouse Nixon to say "Sock It To Me."
The Bold Type [Freeform, 8p]
Last week the Scarlet servers were hacked, leaving journalists scrambling. This week we are heading into a full on flash back to when Sutton, Kat and Jane first started at Scarlet magazine.
Bless This Mess [ABC, 9:30p]
Mike and Rio need a new water heater, but chicken eggs won't cover it…will it?
- HBO proceeds right along in its streak of awesome-looking documentaries — and more often than not, the product matches the flashy promotion. Tonight, they look at the late "Greatest" with Muhammad Ali: What's My Name? and reminds those of us (which is now most of us) who were not alive at the time how controversial he was — which is itself a reminder how messed up America has been and still is.
- ABC's focus on the eventful year that was a half-century ago, 1969, promises a brutal, provocative and important story titled "The FBI & the Panther."
- Tonight's offering of The Last O.G. on TBS sounds somewhat 1970s as a local tough with the comic-genius name BAMF threatens the neighborhood. Tiffany Haddish is also on the Laugh-In special, b tee dubs.
- Finally, a finale; it's classic DC to end the fifth season of The Flash as our title character meets his most polar nemesis, Reverse Flash (will Bizarro Superman make an appearance? Me think probably!)


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"The Mask of Fu Manchu" quotes(1932)
Title The Mask of Fu Manchu
Year 1932
Directors Charles Brabin, Charles Vidor
Genre Sci-Fi, Horror, Adventure
Year 1932
Directors Charles Brabin, Charles Vidor
Genre Sci-Fi, Horror, Adventure
Interpreted by
Plot – An archaeological expedition is searching Genghis Khan tomb. Dr. Fu Manchu wants to take possession of the powerful golden sword and the mask too to dominate the world, but Scotland Yard will stop the criminal.
All actors – Boris Karloff, Lewis Stone, Karen Morley, Charles Starrett, Myrna Loy, Jean Hersholt, Lawrence Grant, David Torrence, Everett Brown, Steve Clemente, Willie Fung, Ferdinand Gottschalkshow all
"The Mask of Fu Manchu" Quotes 8 quotes
"Men of Asia! The skies are red with the thunderbolts of Genghis Khan! They rain down on the white race... and burn them!"
"He'll declare himself Genghis Khan come to life again. And that, my friend, is what you have got to prevent. You must find that grave, and you must be the first to find it."
"I have seen a vision, the prophecy is about to be fulfilled."
"- Nayland Smith: What do you expect to find in that tomb?
- Sir Lionel Barton: A record, the golden plate, the threat of Genghis Khan's return, the golden mask he wore, and the famous golden scimitar."
"Will we ever understand these Eastern races?"


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Knee arthritis affects tens of millions of Americans and can lead to disability. There are quite a few treatments available to help patients avoid knee replacement surgery such as PT, knee injections, PRP therapy, and minimally invasive knee arthroscopy when indicated.
Hear Dr. Farber, Board Certified orthopedic surgeon in Scottsdale and Phoenix, discuss knee arthritis and available treatment options:
Dr. Farber is the Medical Director at Phoenix Shoulder & Knee, offering cutting edge minimally invasive treatments for knee, shoulder and elbow disorders.


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How U.S. Armed Terrorists in Syria
Posted by feww on June 26, 2017
Submitted by a Reader:
95% of terrorists fighting Syrian government are foreigners: Germany's Intelligence Service (BND)
Flood of weapons and entry of 20,000 foreign fighters into Syria "largely defined nature of conflict"
"Three-term Congresswoman Tulsi Gabbard of Hawaii, a member of both the Armed Services and Foreign Affairs committees, has proposed legislation that would prohibit any U.S. assistance to terrorist organizations in Syria as well as to any organization working directly with them. Equally important, it would prohibit U.S. military sales and other forms of military cooperation with other countries that provide arms or financing to those terrorists and their collaborators," a report said.
"Gabbard's 'Stop Arming Terrorists Act' challenges for the first time in Congress a U.S. policy toward the conflict in the Syrian civil war that should have set off alarm bells long ago: in 2012-13 the Obama administration helped its Sunni allies Turkey, Saudi Arabia, and Qatar provide arms to Syrian and non-Syrian armed groups to force President Bashar al-Assad out of power. And in 2013 the administration began to provide arms to what the CIA judged to be 'relatively moderate' anti-Assad groups—meaning they incorporated various degrees of Islamic extremism."
"… as much as 8,000 tons of weapons poured across the Turkish border into Syria just in late 2012 and in 2013."
Weapons were purchased Belgrade, Serbia in May 2013 including "500 Soviet-designed PG-7VR rocket launchers that can penetrate even heavily-armored tanks, along with two million rounds; 50 Konkurs anti-tank missile launchers and 500 missiles, 50 anti-aircraft guns mounted on armored vehicles, 10,000 fragmentation rounds for OG-7 rocket launchers capable of piercing heavy body armor; four truck-mounted BM-21 GRAD multiple rocket launchers, each of which fires 40 rockets at a time with a range of 12 to 19 miles, along with 20,000 GRAD rockets."
"… another Saudi order from the same Serbian company listed 300 tanks, 2,000 RPG launchers, and 16,500 other rocket launchers, one million rounds for ZU-23-2 anti-aircraft guns, and 315 million cartridges for various other guns."
In December 2013 U.S. sold 15,000 TOW anti-tank missiles to the Saudis at a cost of about $1 billion…
"This flood of weapons into Syria, along with the entry of 20,000 foreign fighters into the country—primarily through Turkey—largely defined the nature of the conflict. These armaments helped make al Qaeda's Syrian franchise, al Nusra Front (now renamed Tahrir al-Sham or Levant Liberation Organization) and its close allies by far the most powerful anti-Assad forces in Syria—and gave rise to the Islamic State."
Who told "peaceful demonstrators" to destroy Syrian air defense?
"At the very beginning of the attacks, they [terrorists] started by destroying the Syrian air defense, which has nothing to do with what they called it that time the 'peaceful demonstrations.' Most of the air defense are outside the cities in outlying areas, so they started attacking that air defense and it was affected dramatically during the crisis," said Syrian President Bashar Assad.
One Response to "How U.S. Armed Terrorists in Syria"
June 27, 2017 at 2:57 pm
Are Claims About Syrian Sarin Attacks Propaganda Leading to More War?
BuzzFeed reports: "Syria appears to be preparing a new chemical weapons attack against its citizens, the White House said Monday, warning that if the weapons are again used, the U.S. will make the Syrian government 'pay a heavy price.' … Five U.S. defense officials reached by BuzzFeed News said they did not know where the potential chemical attack would come from, including one U.S. Central Command official who had 'no idea' about its origin."
From RT: "U.S. threats to Syria's legitimate government unacceptable — Kremlin."
See new interview with investigative journalist Seymour Hersh: "Trump Ignored Intel Before Bombing Syria" with The Real News. http://therealnews.com/t2/story:19413:Hersh%3A-Trump-Ignored-Intel-Before-Bombing-Syria
On Sunday, Hersh's piece "Trump's Red Line" was published in the German newspaper Welt am Sonntag.
Hersh wrote: "On April 6, United States President Donald Trump authorized an early morning Tomahawk missile strike on Shayrat Air Base in central Syria in retaliation for what he said was a deadly nerve agent attack carried out by the Syrian government two days earlier in the rebel-held town of Khan Sheikhoun. Trump issued the order despite having been warned by the U.S. intelligence community that it had found no evidence that the Syrians had used a chemical weapon. …
"'The Salafists and jihadists got everything they wanted out of their hyped-up Syrian nerve gas ploy,' the senior adviser to the U.S. intelligence community told me, referring to the flare up of tensions between Syria, Russia and America. 'The issue is, what if there's another false flag sarin attack credited to hated Syria? Trump has upped the ante and painted himself into a corner with his decision to bomb. And do not think these guys are not planning the next faked attack. Trump will have no choice but to bomb again, and harder. He's incapable of saying he made a mistake.'"
THEODORE A. POSTOL, postol at mit.edu
Postol is professor emeritus of science, technology, and national security policy at MIT. Last month, he appeared on an Institute for Public Accuracy news release: "NYT Claims on Syria Attack Unsupported." He said today: "The White House took unjustified actions — and is now creating another set of reasons for more such actions. Chances of an unpredictable escalation are significant. Trump is pushing the Russians to extreme positions and he's undermining the effort to destroy the Islamic State."
FRANCIS BOYLE, fboyle at illinois.edu
Boyle is professor of international law at the University of Illinois College of Law. See his recent interview with Dennis Bernstein at Consortium News: "A Baseless Justification for War in Syria." He said today: "The current posture of U.S., French and perhaps British policy in Syria is basically an invitation for some jihadist group to stage a chemical weapons attack in Syria. That would get blamed on Assad and the U.S. would bomb him. This is an intolerable, incredibly dangerous situation brought on by constant illegal U.S. meddling."


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Holiday home "Am Primusbergerhof" in Bad Goisern on Lake Hallstatt
A place to relax, enjoy some peace and quiet and regain a sense of inner harmony. That is the definition of the Primusbergerhof in Bad Goisern. Peace and a chance to recuperate are guaranteed in our spacious holiday apartments, and of course in our extensive gardens with pet animals and pond.
Your Offers From 28. November To 30. November Change travelling data
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A place to relax, enjoy some peace and quiet and regain a sense of inner harmony. That is the definition of the Primusbergerhof in Bad Goisern.
Peace and a chance to recuperate are guaranteed in our spacious holiday apartments, and of course in our extensive gardens with pet animals and pond.
Our holiday apartment "Armin" sleeps 2-7 persons and has a spacious new kitchen/living area. The bathroom has a shower and WC as well as an anteroom with wardrobe. The apartment is situated on the first floor of a detached building right by the natural bathing area, and is surrounded by a garden and meadow land. The apartment covers an area of 75m² in a quiet location with views of Bad Goisern and the surrounding mountains of the Inner Salzkammergut.
The holiday apartment "Simone" has been available since 2008. With an area of 100m² over 2 floors, the apartment is ideal for 2-8 guests, in very special surroundings. On the ground floor there is a small anteroom with cloakroom and a generously-sized 32m² living room with traditional wood-burning stove and all other fixtures and fittings. This opens on to a new, well-equipped kitchen which opens out on to the outside relaxation area
On the first floor there is a shower and WC as well as two separate bedrooms, each with a double bed.
If you would like to find out more about our apartments, including special offers, please visit our website.
Please do not hesitate to contact us here at the P
- Distance to next ski lift (in km): 20
- Distance to fairground/conference centre (in km): 60
- Outskirts of town
- Distance to next public transport station (in km): 1.5
- Distance to next train station (in km): 3
- Distance to airport (in km): 60
- Rural area
- Distance to city centre (in km): 3
- Distance to the lake (in km): 5
- In the mountains
- Own products
- Pets on request
- Lawns for sunbathing
- Bike storage
- Pets not permitted
- Parking area
- Reduction for children
- Cash only
- No board included
- Number of holiday apartments: 2
- washing machine in the house
- Familial ambience
- Baby chair
- Non-smoking host
- Baby cot
- Internet usage possible
- Comfortable ambience
- Petting zoo
- Garden
- Hiking trails
- Playground
- Table tennis
- Barbecue facilities
- German
- English
- Families
- Elderly persons
- Mountains
- Hiking/nature
- Historical building
- Ski and winter sports
Conditions / Extras
Arrival and departure
- Arrival: from 2.00 pm
- Departure: by 10.00 am
- In the event of cancellation, the terms of the Austrian Hotel Contract Conditions in the current version apply.
- We kindly ask for a deposit of 20% per booking
- Pets are not allowed
- Non-smoking house


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|Echium spp
Wednesday, September 21, 2011
Putting the bubble wrap back
3 comments:
Oh please Margaret, don't decide Winter is on its way yet.ReplyDelete
Today is Autumn Equinox, officially 1st day of Autumn, and i still have dozens of roses, a few hibiscus, some hollyhocks and a few buddliea in bloom.
True, i did put a mulch round my more neglected smaller roses as a protection in case of early frosts, but oh please, not yet.
Do Not worry Ray-here in the North whilst it is true we have on our next layer of undergarments we do not declare it winter until the Rock Shop has closed .ReplyDelete
So warm today that I'm still in T shirts ,even at 9pm ! But had the great shed tidy today ,ready to get the bike in and found a old bench cover for it as I can't ignore the fact that I see daylight through the shed roof ! Now who can I get to sort that out . The whole shed needs replacing ! So ,the easiest solution is to put the electric mower in the garage and ignore the problem . I have placed some strategic buckets . I do hope the mice wont be too damp !ReplyDelete