text:
|Home | About | Journals | Submit | Contact Us | Français
Multiple sclerosis (MS) is a clinically and pathologically heterogeneous inflammatory/demyelinating disease of the CNS. In the MS variant Devic disease, lesions are predominantly found in the optic nerves and spinal cord but not the brain. The immunological bases of the different forms of MS are unknown. We previously generated myelin oligodendrocyte glycoprotein–specific (MOG-specific) TCR transgenic mice (TCRMOG mice; also referred to as 2D2 mice) and reported that a large proportion of these mice develop spontaneous isolated optic neuritis. We have now crossed the TCRMOG mice with MOG-specific Ig heavy-chain knock-in mice (IgHMOG mice; also referred to as Th mice), in which one-third of the B cells are specific for MOG. In these mice, MOG-specific B cells are very efficient in presenting MOG to the transgenic T cells and undergo class switching to IgG1 in the presence of the transgenic T cells. Sixty percent of TCRMOG×IgHMOG mice spontaneously developed a severe form of experimental autoimmune encephalomyelitis (EAE). Histological examination of the CNS revealed a selective distribution of meningeal and parenchymal inflammatory lesions in the spinal cord and optic nerves. Thus, CNS antigen–specific T and B cells cooperate to induce a distinct clinicopathologic EAE pattern that closely replicates human Devic disease.
MS is a clinically and pathologically heterogeneous disease most often characterized by inflammatory and demyelinating lesions throughout the CNS. There are several subtypes of the disease that differ in their CNS lesion distribution (1). Many MS patients first develop isolated optic neuritis, in which lesions are predominantly present in the optic nerves (2, 3). In Asian populations, Devic disease, or neuromyelitis optica (NMO), is the predominant subtype, representing one-third of MS cases (4). Patients with NMO often have a poor prognosis compared with patients with typical MS and develop severe and early relapses (5, 6). Devic disease is characterized by a specific lesion distribution in which inflammatory foci are restricted to optic nerves and the spinal cord with a remarkable absence of lesions in the brain (7). Antibodies (e.g., IgG and IgM) and products of the complement activation cascade have been found in and around CNS lesions, but a pathogenic role for autoantibodies in this disease subtype remains to be formally demonstrated (8). Interestingly, plasmapheresis has been reported to improve the neurologic outcome in patients with severe NMO (9), thereby supporting a role for humoral factors (possibly antibodies) in its pathogenesis.
The cellular and molecular bases for the different subtypes of MS are not well understood, but many features of the human disease have been recapitulated in the EAE animal model. EAE can be induced by immunization of susceptible animals with myelin antigens such as myelin basic protein (MBP), proteolipid protein (PLP), and myelin oligodendrocyte glycoprotein (MOG) (10–12); it is characterized by lymphocytic and mononuclear cell inflammatory infiltrates and demyelination in the brains and spinal cords of affected animals. Clinical manifestations of the disease depend on the antigen and the genetic background of the animal used. Immunization of SJL mice with the PLP aa 139–151 peptide, for example, induces episodes of relapsing-remitting disease in which the mice have a paralytic attack followed by periods of remission (11). On the other hand, immunization of C57BL/6 mice with MOG aa 33–55 produce a chronic form of EAE in which the animals stay paralyzed after their initial attack (12). Therefore each EAE model recapitulates some aspects of the human disease and provides valuable insight into the heterogeneous manifestations of MS.
EAE can be induced by myelin antigen–specific Th1 cells (13, 14); more recently it has been suggested that Th-17 cells are major inducers of EAE (15, 16). By contrast, initial studies with immunization of B cell–deficient mice with the encephalitogenic peptide MOG aa 35–55 suggested that B cells may not be essential for the development of certain forms of EAE (17, 18). However, studies of immunization of B cell–deficient mice with recombinant whole MOG indicated a pathogenic role for B cells in the initiation of EAE (19–21), and other reports suggested that myelin antigen–specific B cells may act as regulatory cells and control the development of EAE (22). Therefore, the role of myelin antigen–specific B cells and antibodies in the pathogenesis of EAE as well as of MS are not well understood.
The result of interactions between myelin antigen–specific B cells and myelin antigen–specific T cells on the development and the phenotype of EAE have not previously been investigated. Here we describe a mouse EAE model in which both T and B cells are specific for the same autoantigen MOG. The presence of MOG-specific B and T cells in the peripheral repertoire resulted in the development of severe and spontaneous EAE in the majority of the mice. Histopathological analysis of the CNS of the affected mice showed a preponderance of inflammatory lesions in the leptomeninges and parenchyma of the spinal cords and optic nerves but not in the brains of affected mice. Our results suggest that an active cooperation between MOG-specific T and B cells results in a spontaneous and severe form of EAE with a lesion distribution pattern typical of human Devic disease.
We previously generated MOG-specific TCR transgenic mice on a C57BL/6 background (TCRMOG mice; also referred to as 2D2 mice) in which the transgenic TCR-bearing cells are efficiently selected in the thymus and are seeded to the peripheral immune compartment (23). Although over 95% of the peripheral T cells express transgenic TCR, only a small percentage (4%) of TCRMOG mice develop spontaneous EAE. Nevertheless, a significant number of the transgenic mice develop spontaneous optic neuritis (47%). Immunization of TCRMOG mice with the MOG peptide in complete Freund's adjuvant plus pertussis toxin resulted in the development of full-blown EAE, suggesting that EAE can be induced in the transgenic mice if there is appropriate activation of transgenic TCRMOG T cells. Since antigen-specific B cells can be efficient APCs, we hypothesized that MOG-specific B cells could activate TCRMOG T cells and induce spontaneous EAE in the TCRMOG mice. Therefore, we crossed the TCRMOG mice with MOG-specific Ig heavy-chain knock-in mice on a C57BL/6 background (IgHMOG mice; also referred to as Th mice), in which 20% of peripheral B cells are specific for the autoantigen MOG (24), thus providing B and T cells specific for the same MOG antigen in the peripheral repertoire of the mice.
To determine whether the presence of MOG-specific B cells could change the T cell response of TCRMOG T cells in TCRMOG×IgHMOG mice, we first analyzed the proliferation of splenocytes from TCRMOG×IgHMOG and control TCRMOG mice crossed with nitrophenyl-specific (NP-specific) Ig heavy-chain knock-in mice on a C57BL/6 background (TCRMOG×IgHNP mice; IgHNP also referred to as B1.8 mice) in response to MOG aa 35–55 peptide or whole recombinant MOG aa 1–125 (rMOG) protein. Splenocytes from TCRMOG×IgHMOG mice, carrying MOG-specific T and B cells, and TCRMOG×IgHNP mice, carrying MOG-specific T cells and NP-specific B cells, proliferated equally well in response to MOG aa 35–55 peptide (Figure (Figure1A).1A). As a control, splenocytes from IgHMOG mice, IgHNP mice, and OVA-specific TCR transgenic mice on a C57BL/6 background (TCROVA mice; also referred to as OT-II mice) crossed with IgHMOG mice (TCROVA×IgHMOG mice), all lacking the MOG-specific T cells, did not respond to MOG aa 35–55 peptide (Figure (Figure1A).1A). Similarly, splenocytes from IgHNP, IgHMOG, or TCROVA×IgHMOG mice did not proliferate in response to whole rMOG (Figure (Figure1A).1A). However, splenocytes from TCRMOG×IgHMOG mice, which express MOG-specific T and B cells, proliferated more strongly and at lower concentrations to rMOG than the control TCRMOG×IgHNP mice (Figure (Figure1A). 1A).
The enhanced response of TCRMOG×IgHMOG splenocytes to rMOG could be attributed to the proliferation of either T or B cells or both. To determine which cell type was responsible for the enhanced proliferation, we first tested the response of T cells in the presence of B cells of different specificities. We initially separated T cells from the spleens of TCRMOG mice and stimulated them with purified B cells from either IgHNP or IgHMOG mice together with MOG aa 35–55 peptide or rMOG protein as a source of specific antigen. Whereas MOG aa 35–55 peptide induced a strong proliferation of MOG-specific T cells independently of the presence of specific B cell populations, whole rMOG protein led only to dose-dependent proliferation in the presence of MOG-specific B lymphocytes (Figure (Figure1B).1B). To confirm that the observed response corresponded to T cell proliferation, we stimulated CFSE-labeled splenocytes of TCRMOG mice in the presence of MOG-specific B cells (IgHMOG) or NP-specific B cells (IgHNP) and different concentrations of MOG aa 35–55 peptide or rMOG and measured T cell proliferation by CFSE dilution on CD4+ T cells. Using this approach we observed similar numbers of dividing TCRMOG T cells in response to MOG aa 35–55 peptide independent of the antigen specificity of the B cells used as APCs (Figure (Figure1C).1C). In contrast, more TCRMOG T cells divided in response to rMOG in the presence of IgHMOG MOG-specific B cells than in the presence of IgHNP NP-specific B cells, confirming that provision of MOG-specific B cells to T cells of the same specificity enhanced their proliferative response (Figure (Figure1C1C and Supplemental Figure 1; supplemental material available online with this article; doi:10.1172/JCI28334DS1).
Next we analyzed T cell cytokines secreted under these culture conditions. Stimulation of TCRMOG T cells with rMOG and B cells led to a dose-dependent secretion of IFN-γ. IFN-γ production in response to rMOG was greater in the presence of MOG-specific IgHMOG B cells than in the presence of NP-specific B cells (Figure (Figure1D).1D). Intracellular anti–IFN-γ staining of CD4+ T cells confirmed that MOG-specific CD4+ T cells were responsible for the increased IFN-γ production observed (data not shown). In contrast, IFN-γ levels were comparable among T cells stimulated with MOG aa 35–55 peptide, IgHMOG B cells, and IgHNP B cells (Figure (Figure1D).1D). These data suggest that the MOG protein is selectively processed (25) and presented by MOG-specific B cells, resulting in more efficient activation of MOG-specific T cells when both cell types are present in the peripheral repertoire. The Th2-associated cytokines IL-4 and IL-10 and the pathogenic cytokine IL-17, which is the signature of Th-17 cells, could not be detected in the supernatants (data not shown).
Next we analyzed the role of B cells in the enhanced response of TCRMOG×IgHMOG splenocytes to rMOG. B cells were purified from either IgHMOG (MOG-specific) mice or IgHNP (NP-specific) mice, labeled with CFSE, and cultured in the presence of purified T cells from either TCRMOG (MOG aa 35–55–specific) mice or TCROVA (OVA aa 323–339–specific) mice. B cell proliferation was evaluated by CFSE dilution on B220+ B cells, and different cell surface activation molecules were assessed by flow cytometry analysis. Provision of MOG protein to MOG-specific B cells was not sufficient to trigger B cell division (Figure (Figure2A).2A). Coculture with activated TCRMOG T cell lines and MOG led to a dose-dependent proliferation of MOG-specific IgHMOG B lymphocytes (Figure (Figure2A)2A) and upregulation of activation markers such as CD69, CD80, and CD86 (Supplemental Figure 2 and data not shown). However, MOG-specific B cells also proliferated upon coculture with activated OVA-specific T cell lines and rMOG, as did NP-specific B cells upon coculture with activated MOG-specific T cells (Figure (Figure2B).2B). These results show that, in vitro, the presence of activated T cells alone rather than antigen-specific T cells is sufficient for the induction of B lymphocyte proliferation.
We previously reported that very few (4%) TCRMOG mice develop spontaneous EAE, but a significant portion develops isolated optic neuritis (23). We also found that the development of full-blown EAE in TCRMOG mice was in part dependent on the immunization regimen, suggesting that isolated optic neuritis may progress to EAE if there is appropriate activation of MOG-specific T cells. Based on our in vitro data indicating that antigen-specific B cells efficiently activated MOG-specific T cells, we followed the development of spontaneous EAE in TCRMOG×IgHMOG and control mice.
Consistent with previous reports, no spontaneous EAE was observed in IgHMOG mice (24), and TCRMOG mice developed a very low incidence of spontaneous EAE (6%; Table Table1).1). The presence of transgenic T cells of a different specificity (OVA specific) also did not result in the development of spontaneous EAE in TCROVA×IgHMOG mice (Table (Table1).1). In sharp contrast, 59% of TCRMOG×IgHMOG mice carrying MOG-specific T and B cells developed severe EAE (Table (Table1).1). The disease had an abrupt onset with rapid progression to a clinical score of 2.5–3 over 1–2 days in most mice with disease (Supplemental Figure 3). We did not observe any gender bias for the development of spontaneous disease in TCRMOG×IgHMOG mice. Incidence as well as mean clinical scores among TCRMOG×IgHMOG mice were higher than in TCRMOG mice with spontaneous EAE. Spontaneous disease was also accelerated in that it developed earlier in TCRMOG×IgHMOG mice (44.1 days after birth) than in TCRMOG mice (73.5 days after birth). Mortality among TCRMOG×IgHMOG mice was 10% (3 of 29), compared with 0% (0 of 5) in TCRMOG mice. The severity of the disease was also apparent histologically, as TCRMOG×IgHMOG mice had a mean of 188.4 CNS inflammatory foci compared with 16.6 in the TCRMOG mice (Table (Table1).1). No histologic disease was observed in TCRMOG×IgHMOG mice without clinical disease, TCRMOG×IgHNP mice, or IgHMOG mice (Table (Table1).1). These results suggested that the presence of MOG-specific B cells is crucial for increased incidence and severity of spontaneous EAE in TCRMOG mice.
We determined the anatomic localization of inflammatory lesions in the different areas of the CNS of TCRMOG and TCRMOG×IgHMOG with EAE. In TCRMOG mice with EAE, most of the lesions (40%) were localized in the cerebral hemispheres (Table (Table2).2). The remaining foci were equally distributed among the midbrain-brainstem, the cerebellum, and the spinal cord. These mice also had optic neuritis. In contrast, most of the lesions of TCRMOG×IgHMOG mice were localized in their spinal cords and optic nerves, with very few in the brain (less than 3% in cerebral hemispheres, midbrain-brainstem, or cerebellum; Table Table22 and Figure Figure3A).3A). This lesion distribution pattern is similar to that of Devic disease. This selective distribution of inflammatory foci in the spinal cords and optic nerves was observed in 100% of the affected TCRMOG×IgHMOG mice analyzed histologically (Table (Table1),1), but not in TCRMOG mice with EAE. The inflammatory lesions were composed of lymphocytes and monocytes and were present in both leptomeninges and parenchyma, particularly the subpial regions where there was marked associated demyelination and edema (Figure (Figure3A).3A). Peripheral nerve roots were intact. Phenotypic analysis on cryosections demonstrated infiltration of T cells (CD4+ and TCR Vβ11+) and large numbers of B cells (B220+; Figure Figure3B).3B). The accumulations of lymphocytes were particularly marked in the subarachnoid spaces around the spinal cord of TCRMOG×IgHMOG mice, and the aggregates focally formed follicle-like structures that were made evident using reticulin stains (Figure (Figure3C).3C). Similar patterns of germinal center–like structures have previously been described in the leptomeninges of patients with MS (26, 27). We did not identify substantial numbers of eosinophils or neutrophils in these lesions. Collectively, these data indicate that TCRMOG×IgHMOG mice have a selective Devic-like distribution of inflammatory lesions, i.e., in the spinal cord and the optic nerves, which is characterized by a close and well-organized follicule-like architecture that may allow close interaction between antigen-specific T and B cell populations in the target tissue.
To obtain insight into the mechanisms responsible for the development of Devic-like disease in TCRMOG×IgHMOG mice, we analyzed the phenotype of both T and B cells ex vivo. Between 6 and 8 weeks of age, CD4+ T cells from TCRMOG mice were mostly naive, and only 1.8% of them were activated/memory cells in the spleen (Figure (Figure4A).4A). Healthy TCRMOG×IgHMOG mice had similar numbers of activated cells (2.4%). In contrast, an increased proportion of CD4+ T cells (4.7% and 8.2% in spleens and lymph nodes, respectively) from diseased TCRMOG×IgHMOG mice were activated and expressed CD69 (Figure (Figure4A).4A). Analysis of the number of regulatory cells, as determined by Foxp3 intracellular staining in CD4+ T cells, showed a small but comparable number of CD4+Foxp3+ T cells (2%) in the spleens and lymph nodes of healthy and diseased TCRMOG×IgHMOG mice (data not shown).
Next we looked at the expression of different markers on the surface of B cells from the various mouse strains. We did not find any difference in the expression of MHC class II molecules, B7.2, or IgMa on the surface of B cells from IgHMOG or healthy or diseased TCRMOG×IgHMOG mice (Figure (Figure4B4B and data not shown). However, we observed an increase in the percentage of B220+B7.1+ cells in the spleens of TCRMOG×IgHMOG mice with disease compared with healthy TCRMOG×IgHMOG mice. The upregulation of the B7.1 molecule on the surface of B cells in TCRMOG×IgHMOG mice with Devic-like disease suggests that these B cells have been activated. To determine whether MOG-specific B cells differentiated into effector B cells, we measured levels of MOG-specific antibodies in serum samples of IgHMOG, TCRMOG, TCRMOG×IgHMOG, TCROVA×IgHMOG, and TCRMOG×IgHNP mice. No MOG-specific antibodies were detected in the absence of IgHMOG MOG-specific B cells, i.e., in TCRMOG mice and in TCRMOG×IgHNP mice. Without any immunization, IgHMOG mice and TCROVA×IgHMOG mice produced large amounts of MOG-specific IgM, IgG2a, and IgG2b antibodies but low levels of IgG1 (Figure (Figure4C).4C). In contrast, serum samples from TCRMOG×IgHMOG mice, in which MOG-specific T cells and B cells are present in the peripheral repertoire, contained lower amounts of MOG-specific IgM (P = 0.04) but higher amounts of MOG-specific IgG1 antibodies (P = 0.03) than the IgHMOG and TCROVA×IgHMOG mice. MOG-specific antibody isotype switching to IgG1 was consistently found in all TCRMOG×IgHMOG mice analyzed, whether they had clinical EAE or were asymptomatic (Figure (Figure4C).4C). Isotype switching was not dependent on age, since serum samples obtained from 4-week-old and adult TCRMOG×IgHMOG mice showed high levels of anti-MOG IgG1 antibodies.
To further characterize the MOG-specific T cell responses involved in the development of Devic-like disease, we purified mononuclear cells from WT, IgHMOG, and healthy and diseased TCRMOG×IgHMOG mice. Consistent with the absence of disease in the WT, IgHMOG, and healthy TCRMOG×IgHMOG mice, there were small numbers of mononuclear cells present in the CNS of these animals (data not shown). In these 3 strains of mice, CD4+ T cells represented 3% of the few infiltrating monocytes. However, in TCRMOG×IgHMOG mice with Devic-like disease, there was an increase in the number of infiltrating monocytes (data not shown) and a 5-fold increase in the percentage of CD4+ T cells in the CNS tissues (Figure (Figure5A).5A). Since both Th1 and Th-17 cells have been implicated in the development of EAE (15, 16, 28, 29), we determined whether T cells from TCRMOG×IgHMOG mice with EAE could secrete these cytokines. Although IL-17 production was not detected in our in vitro cultures (data not shown), intracellular cytokine staining of CD4+ T cells obtained from the CNS of TCRMOG×IgHMOG mice showed that CD4+ T cells involved in the development of Devic-like disease secreted IFN-γ and IL-17 (Figure (Figure5B).5B). Since only 4% of TCRMOG mice develop spontaneous EAE, it has not been possible to obtain CNS samples from these animals to perform FACS analysis together with those of TCRMOG×IgHMOG mice with disease. However, we have prepared RNA from the spinal cords of TCRMOG×IgHMOG mice with Devic-like disease and TCRMOG mice with spontaneous EAE and analyzed the expression of IL-17 mRNA by real-time PCR. This analysis showed that there was an increase in expression of both IL-17 and IFN-γ mRNA in the spinal cords of diseased TCRMOG×IgHMOG mice compared with TCRMOG mice with EAE (Figure (Figure5C).5C). Although we could not determine which cell type was responsible for the increased IL-17 and IFN-γ production observed in the spinal cords of TCRMOG×IgHMOG mice, together with the intracellular cytokine staining, these data suggest that more Th1 and Th-17 cells are infiltrating the spinal cords of TCRMOG×IgHMOG mice than do so in TCRMOG mice.
While the enhanced and selective activation of TCRMOG T cells in the presence of MOG-specific B cells can explain the high incidence of spontaneous disease observed in TCRMOG×IgHMOG mice compared with TCRMOG mice, it is not clear why TCRMOG×IgHMOG mice have Devic-like disease and not typical EAE. To begin to address this issue, we compared the expression of the autoantigen MOG in different CNS regions of C57BL/6 mice. Consistent with our previous observation (23), we found that MOG mRNA was more abundant in the optic nerve than the spinal cord (Figure (Figure6).6). We also found that MOG mRNA was more abundant in the spinal cord than in the brain. This selective distribution of MOG mRNA might contribute at least in part to the preferential localization of lesions in the spinal cords and optic nerves of TCRMOG×IgHMOG mice.
Here we report the generation of what we believe to be a new mouse model in which both T and B cells are specific for the same myelin antigen, MOG. We demonstrated that there was active cooperation between T and B cells in these animals: the presence of MOG-specific T cells led to massive production of MOG-specific IgG1 antibody, and MOG-specific B cells also enhanced MOG-specific T cell proliferation and activation. Strikingly, this cooperation resulted in the development of a spontaneous and severe form of EAE characterized by the specific presence of inflammatory foci in the spinal cords and optic nerves of the animals. This lesion distribution pattern is very similar to that of human NMO.
B lymphocytes have been implicated in several autoimmune diseases, mostly because of their capacity to produce specific autoantibodies. B cells and plasma cells are often present in MS lesions (30, 31), and oligoclonal Ig bands have been observed in the cerebrospinal fluid of many but not all MS patients (32, 33). The presence of Ig and products of complement activation cascade deposited around the vessels suggests a pathogenic role for autoantibodies in NMO (8). In addition, a serum IgG autoantibody (NMO-IgG) serves as a specific marker for NMO (34). Recent work has shown that the NMO-IgG is not specific for myelin antigens but selectively binds to the aquaporin-4 water channel, a component of the dystroglycan protein complex located in astrocytic foot processes at the blood-brain barrier (35).
The high expression of class II MHC and costimulatory molecules by activated B cells suggest that they could also play a role in driving the T cell responses as a result of antigen presentation to large numbers of antigen-specific T cells (36, 37). In vivo, depletion of B cells from normal mice can reduce the magnitude of the immune response, suggesting that B cells contribute to the T cell response (38–41), although primary T cell responses can also develop in B cell–deficient mice (42). Therefore, a number of reports suggested that antigen-specific B cells induce naive CD4+ T cell proliferation in vivo (43–45). The importance of B cells as APCs in some autoimmune diseases such as proteoglycan-induced autoimmune arthritis has recently been demonstrated (46). In contrast with this arthritis model, EAE is generally considered to be mainly T cell driven (28). Nevertheless, it is important to note that B lymphocytes are required for the induction of EAE when MOG protein, but not MOG peptide, is used for the induction of EAE (19). Using a double antigen-specific transgenic model, we demonstrate here that, in contrast to wild-type splenocytes containing the classical APCs, MOG-specific B cells were able to stimulate the proliferation of MOG-specific T cells in the presence of a limited amount of rMOG. Since IgHMOG B cells only recognize conformationally dependent epitopes of MOG but not MOG aa 35–55 (47), the enhanced T cell response in the presence of IgHMOG B cells and MOG most likely resulted from the capture of MOG through B cell surface Ig that caused enhanced presentation of the antigen rather than direct presentation of MOG aa 35–55 epitope.
Ex vivo analysis of T cells from TCRMOG×IgHMOG mice revealed that unless mice exhibited clinical disease, there was no marked increase in the number of activated T cells in the periphery (Figure (Figure4).4). Development of Devic-like disease correlated with the infiltration of both T and B cells into the spinal cord (Figure (Figure3B3B and Figure Figure5A).5A). The disease was also associated with the presence of lymphoid follicle–like structures in the spinal cords of TCRMOG×IgHMOG mice with Devic-like disease, suggesting an active cooperation between T and B cells in situ (Figure (Figure3C).3C). Similar to actively induced EAE, we observed that both IL-17– and IFN-γ–producing T cells infiltrated the CNS of TCRMOG×IgHMOG mice with Devic-like disease. Because of the low frequency of spontaneous disease observed in the TCRMOG mice, it was not possible to perform parallel intracellular cytokine staining of CD4+ T cells infiltrating the brains of TCRMOG mice with spontaneous EAE. However, IL-17 mRNA expression was greater in the spinal cords of TCRMOG×IgHMOG mice with Devic-like disease than in TCRMOG mice with EAE. Therefore the increase in IL-17 expression in the CNS of TCRMOG×IgHMOG mice with Devic-like disease might reflect an increase in the number of pathogenic Th-17 cells. Alternatively, we cannot exclude the possibility that other cell types might be responsible for the enhanced IL-17 production. The role of the CNS cytokine milieu and MOG-specific B cells in the differentiation of Th-17 cells is currently under investigation.
Th cells are important for the differentiation of B cells into Ig-secreting plasma cells or long-lived memory B cells. In addition, interaction between antigen-specific T and B cells is required for B cell maturation and Ig isotype class switching. Consistent with the role played by Th cells in the maturation of B cells, we also observed that B cells from TCRMOG×IgHMOG mice underwent class switching from IgM to IgG1 in vivo. Ig switch is dependent on the subset of T cells providing help to B cells, and IgG1 is regarded as a Th2-dependent isotype (48). In TCRMOG×IgHMOG mice, however, the T cell response seems to be biased toward Th1 cells, since we only detected IFN-γ and not Th2 cytokines in cultures of TCRMOG or TCRMOG×IgHMOG mouse spleen cells. In addition, we observed an increase in the number of B7.1-expressing B cells in TCRMOG×IgHMOG mice with Devic-like disease. B7.1 is a costimulatory molecule associated with enhanced Th1 responses. Therefore, the requirement for Th2 responses in IgG1 class switching might not be absolute in this model. In support of this hypothesis are previously published results showing that mice deficient in the Th2-specific cytokine IL-4 have diminished but still robust production of IgG1 (48). Alternatively, other factors present in the microenvironment might contribute to the IgG1 class switching observed in TCRMOG×IgHMOG mice. Taken together, the TCRMOG×IgHMOG model provides an interesting and, to the best of our knowledge, unique model to study the induction and cooperation of effector autopathogenic T and B cell responses in vivo and in autoimmunity.
In TCRMOG×IgHMOG mice it is unlikely that MOG-specific antibodies themselves are responsible for the initiation of spontaneous Devic-like disease, since we did not observe a substantial difference in the level of MOG-specific antibodies between healthy and diseased TCRMOG×IgHMOG mice (Figure (Figure4C),4C), and the transfer of serum from TCRMOG×IgHMOG mice to TCRMOG mice did not induce EAE or Devic-like disease (data not shown). However, the combination of enhanced antibody production and a robust MOG-specific pathogenic T cell response might be responsible for the fulminant disease observed in TCRMOG×IgHMOG mice. In addition, the antibody and T lymphocyte specificity might play a role in determining the specific lesion distribution pattern as follows: First, the amount of antigen might be important. Previously we reported that there is relatively more MOG expressed in the optic nerves than in the spinal cords of C57BL/6 mice (23). Here we additionally demonstrated that mRNA for MOG was more abundant in the spinal cords than the brains of these mice. It is therefore possible that a differential expression of MOG protein results in predominant attack of optic nerves and spinal cords in TCRMOG×IgHMOG mice because there are relatively higher levels of the target antigen. Second, the accessibility of the autoantigen might be a critical factor. While the white matter in the spinal cord is on the outside and bathed in the cerebrospinal fluid, the gray matter is on the outside in the cerebellum and cerebrum. In the TCRMOG×IgHMOG mice, the inflammatory infiltrates in the subarachnoid space surrounding the spinal cord and around the optic nerves were more numerous than generally observed in induced EAE models and TCRMOG mice with spontaneous EAE. Therefore, because MOG antigen might be more accessible in the spinal cords and optic nerves, enhanced MOG-specific T and B cell responses generated in TCRMOG×IgHMOG mice might preferentially attack these CNS areas, resulting in selective lesion distribution similar to that of NMO.
Autoimmune diseases have a complex pathogenesis, and the TCR transgenic mice have proven to be valuable tools to investigate the behavior of self-reactive T cells, their interactions with their antigenic targets, and their induction of disease. Several TCR transgenic mice specific for autoantigens have been generated, and many of the autoantigen epitopes recognized by transgenic T cells have restricted expression in the CNS (49). Despite the striking increase in the frequency of self-reactive T cells in most of the TCR transgenic mouse lines, however, it is noteworthy that spontaneous autoimmune diseases do not occur in 100% of the animals. For example, 60% of PLP TCR transgenic mice develop spontaneous EAE (50), and the incidence of spontaneous EAE in MBP aa 1–11–specific TCR transgenic mice ranges from 0% to 14% depending on the line (51, 52). While 47% of TCRMOG mice develop spontaneous clinical optic neuritis, only 4% of these animals develop spontaneous EAE (23). The initiation of disease most likely requires adequate priming of the autoimmune response and/or control of regulatory T cell responses. In one of the MBP-specific TCR transgenic lines, spontaneous disease occurs in 15% of the animals housed in a pathogen-free facility and 43% of those housed in conventional facilities (51), suggesting a role for infectious agents as triggers of the disease. At the present time, the environmental factor(s) in specific pathogen–free facilities that may initiate spontaneous EAE and are responsible for differences in incidence of spontaneous EAE remain unidentified. Moreover, the mechanisms underlying initiation of spontaneous EAE by infectious agents remain to be elucidated. The inflammatory response generated by infection could trigger the autoreactive T cell response and initiate autoimmune diseases. Alternatively, molecular mimicry between microbial and self antigens has been proposed as a mechanism for the development of EAE (53). Of particular relevance in this regard to the present study is the fact that there are a large number of molecular homologs of the extracellular domain of MOG. Specifically, the butyrophilin gene products exhibit linear aa sequence homologies with MOG ranging from 35% to 50% and have in some cases been shown to exhibit molecular mimicry with MOG (54). In TCRMOG×IgHMOG mice, autoreactive MOG-specific T cells may recognize an antigen mimicking MOG that is present in the peripheral immune compartment and then get further activated and amplified by MOG presented by large numbers of MOG-specific B cells. This might then lead to a high frequency of spontaneous disease. Since the restricted expression of MOG to the CNS has recently been challenged (55), it is also possible that MOG itself expressed in the periphery or released into the peripheral circulation could be presented to autoreactive T cells by MOG-specific B cells very efficiently. Alternatively, the increased incidence of spontaneous EAE observed in MBP TCR transgenic mice crossed to RAG-deficient mice and therefore lacking regulatory T cells expressing endogenous TCR shows the importance of regulatory cells in keeping autoreactive T cells in check (49). Since only 4% of TCRMOG mice and 60% of TCRMOG×IgHMOG mice develop spontaneous EAE (23), we compared the number of regulatory cells between these 2 strains. Although we found that the number of CD4+Foxp3+ T cells was low in TCRMOG mice (around 2%), it was similar in TCRMOG×IgHMOG mice. Additional experiments are necessary to determine whether B cells can modulate the activity of regulatory cells.
In summary, TCRMOG×IgHMOG mice represent a useful model to determine the factors responsible for different lesion localization and to study the role of T cell/B cell cooperation in the induction of autoimmune pathologies. In addition, since TCRMOG×IgHMOG mice develop a high incidence of spontaneous disease involving both T and B cells, these mice may be useful for testing the biology of T cell/B cell cooperation in the genesis of autoimmune diseases and for testing novel therapies for MS and Devic disease.
C57BL/6 mice transgenic for a TCR with specificity for the peptide MOG aa 35–55 (TCRMOG mice; also referred to as 2D2 mice) were extensively described previously (23). As control for TCRMOG mice, we used TCROVA mice (also referred to as OT-II mice), which are transgenic for a TCR recognizing the peptide OVA aa 323–339 (The Jackson Laboratory).
IgHMOG mice (also referred to as Th mice), which contain MOG-specific B cells, were generated by replacing the germline JH locus with the rearranged Ig heavy-chain (IgH) variable (V) gene of a pathogenic MOG-specific monoclonal antibody (24). The B cells exclusively express the transgenic H chains, and one-third bind MOG. As control for the IgHMOG mice, we used NP-specific IgHNP mice (also referred to as B1.8 mice; ref. 56).
Mice were housed in a specific pathogen–free, viral antibody–free animal facility at the Harvard Institutes of Medicine. All breeding and experiments were reviewed and approved by the Institutional Animal Care and Use Committee of Harvard Medical School. Serums from 4- to 10-week-old mice were obtained at sacrifice by cardiac puncture.
MOG aa 35–55 peptide (MEVGWYRSPFSRVVHLYRNGK) and OVA aa 323–336 peptide (ISQAVHAAHAEINE) were obtained from the Biopolymer Facility, Center for Neurological Diseases, Brigham and Women's Hospital. A plasmid construct encoding the extracellular domain of rat MOG protein (MOG aa 1–125) was generously provided by C. Linington (University of Aberdeen, Aberdeen, United Kingdom), and rMOG protein was purified from inclusion bodies (57). OVA was purchased from Sigma-Aldrich.
T and B lymphocytes were isolated from spleens and lymph nodes using anti-CD4 and anti-B220 columns, respectively, according to the manufacturer's instructions (Miltenyi Biotec). Purity of the positively selected cell populations was >95% as assessed by flow cytometry. For the analysis of B cell proliferation and activation in the presence of antigen-specific T cells, we used T cell lines obtained by 2 rounds of stimulation of lymphocytes from TCR transgenic mice with 10 μg/ml MOG aa 35–55 or OVA aa 323–326 in the presence of irradiated splenocytes and 20 U/ml IL-2.
Cells were grown in RPMI supplemented with 10% FCS, β-mercaptoethanol, l-glutamine, gentamicin sulfate, and penicillin/streptomycin. For thymidine proliferation assay, either 5 × 106 cells/ml total splenocytes or 2.5 × 105 cells/ml purified CD4+ cells plus 2.5 × 106 cells/ml purified B220+ B cells were cultured for 72 hours in 96-well plates in the presence of different concentrations of the rMOG protein or MOG aa 35–55 peptide (range, 0.01–100 μg/ml). Cells were pulsed with 1 μCi 3H-thymidine for the last 16 hours of incubation. Mean thymidine incorporation in triplicate wells was measured using a β counter (LS 5000; Beckman Coulter).
For the assessment of proliferation by CFSE dilution assay, the cells were labeled with CFSE (Invitrogen) at 2 μM in PBS, washed, and cultured at 5 × 105 cells/ml for 2–4 days according to the specific stimulation protocol. Cells were then harvested and analyzed by flow cytometry.
Unlabeled or CFSE-labeled cells were blocked with antibodies to the FcγIII/II receptors (CD16/CD32) to avoid nonspecific staining and were subsequently labeled at 4°C with the following monoclonal antibodies (BD Biosciences): anti-CD23–PE, anti-CD40L–PE, anti-CD69–PE, anti-CD80–PE, anti-CD86–PE, anti-IgM–PE, anti-IgG1–PE, anti-IgG2a–PE, anti-IgG2b–PE, anti-IgG3–PE, anti-CD4–allophycocyanin, and anti-B220–allophycocyanin. Isotype- and concentration-matched control antibodies were used to assess nonspecific staining. Cells were then fixed in 0.3% paraformaldehyde and kept at 4°C until analysis.
For the detection of intracellular cytokines, the cells were restimulated for 5 hours with PMA (50 ng/ml) and ionomycin (1 μM), in the presence of 3 μM monencin for the last 3 hours. The cells were then stained for surface markers and fixed. Subsequently, the cells were permeabilized with saponin and stained with PE-labeled anti-cytokine antibodies resuspended in saponin buffer.
The cells were analyzed on a 4-color FACSort (BD Biosciences), and lymphocytes were gated on forward and side scatter as well as on CD4 and B220 for T and B cells, respectively.
IL-2, IL-4, IL-10, and IFN-γ concentrations were determined in cell culture supernatant by a standard sandwich ELISA by coating 96-well plates with the appropriate anti-cytokine monoclonal antibodies (BD Biosciences), incubating the plates with the culture supernatant overnight, and detecting the bound cytokines with a second biotinylated anti-cytokine antibody and avidin peroxidase. Results were reported in pg/ml according to a standard dilution curve.
For measuring MOG-specific antibodies in serum, 96-well plates were coated with 4 μg/ml MOG in PBS, blocked, and incubated with sera (1:40 and 1:200 dilution for IgM and 1:200 and 1:1,000 dilution for IgG isotypes) for 2 hours. After washing, the serum antibodies retained by the plate-bound MOG were detected by biotinylated, isotype-specific anti-Ig antibodies (BD Biosciences) and avidin peroxidase. Nonspecific binding was assessed using OVA-coated plates as negative control. Results were reported as OD.
Clinical assessment of EAE was performed daily, and mice were scored for disease according to the following criteria: 0, no disease; 1, decreased tail tone; 2, hind-limb weakness or partial paralysis; 3, complete hind-limb paralysis; 4, front- and hind-limb paralysis; 5, moribund state.
Animals were sacrificed after 3 months of observation for development of clinical signs of EAE or after at least 2 weeks of EAE. Brains, spinal cords, and optic nerves were removed and fixed in 10% formalin. Paraffin-embedded sections were stained with hematoxylin and eosin, Luxol fast blue, and reticulin and by Bielschowsky preparation for light microscopy. Immunohistochemistry was performed on cryosections using antibodies to CD4, TCR Vβ11, and B220.
Ig serum levels in TCRMOG×IgHMOG mice versus TCRMOG or IgHMOG control mice were compared by 2-tailed Student's t test. P values less than 0.05 were considered to be statistically significant.
We thank Hartmut Wekerle, Andreas Holz, and Gurumoorthy Krishnamoorthy for providing IgHMOG mice; Klaus Rajewsky for providing IgHNP mice; Christopher Linington for providing the soluble MOG-encoding plasmid; and Michael Nazareno for assistance with screening of mice. This work has been supported by grants from the NIH (NS046414 to R.A. Sobel and NS38037, AI44880, NS30843, and NS45937 to V.K. Kuchroo and E. Bettelli) and the National MS Society (NIH grant RG2571 to V.K. Kuchroo and E. Bettelli). V.K. Kuchroo is a recipient of a Javitz Neuroscience Investigator Award from the NIH.
Nonstandard abbreviations used: IgHMOG mice, MOG-specific Ig heavy-chain knock-in mice on a C57BL/6 background; IgHNP mice, NP-specific Ig heavy-chain knock-in mice on a C57BL/6 background; MBP, myelin basic protein; MOG, myelin oligodendrocyte glycoprotein; NMO, neuromyelitis optica; NP, nitrophenyl; PLP, proteolipid protein; rMOG, recombinant MOG aa 1–125; TCRMOG mice, MOG-specific TCR transgenic mice on a C57BL/6 background; TCROVA mice, OVA-specific TCR transgenic mice on a C57BL/6 background.
Conflict of interest: The authors have declared that no conflict of interest exists.
Citation for this article: J. Clin. Invest. 116:2393–2402 (2006). doi:10.1172/JCI28334.
Estelle Bettelli and Dominique Baeten contributed equally to this work.
See the related Commentary beginning on page 2313.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's Canada Institute for Scientific and Technical Information in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


text:
Inkkas Spectrum Low Top Shoes
Item # 239121
Take An Additional 25% Off
Stash Points: 3,498
What is this?This is the number of points you get in The Zumiez Stash for purchasing this item. Stash points are redeemable for exclusive rewards only available to Zumiez Stash members. To redeem your points check out the rewards catalog on thestash.zumiez.com.
Put some color in your life with this handmade low top shoes crafted with an authentic textile upper, and for every pair purchased Inkkas will plant a tree.
|More Details

subset: commoncrawl


text:
From the post How to identify if a webpage is being loaded inside an iframe or directly into the browser window? I understand you can detect whether or not your page is being viewed through a frame or not.
Using a WordPress site. I'd like to append a body class tag to my site dependent on it's view. I'm doing this so I can tweak the styling. I understand this can't be done via PHP and you need to use JavaScript.
Can somebody walk me through an example of how I can add my body class with this?
if (top === self) { not iframe -do nothing } else { in a frame - add body class }
Thanks in advance for the hand holding.


text:
Is your resume in need of a little disruption?
Startups are all about
making things work better.
Our resume writing service is all about doing that for your CV.
StartUpHire is the place to find Marketing jobs in and near Alameda. Review the list below of Marketing Alameda jobs. These Marketing jobs represent some of the most rewarding careers available. You can refine your job search for Marketing jobs in Alameda by adding a job title, company name, or keyword to the search box above … or click the "Advanced Search" link for more options. Don't see the employment opportunity you're looking for? Check back soon for more Marketing jobs in Alameda, or sign up for an email alert (link in upper left) to receive periodic updates on new Marketing jobs in Alameda, CA.
Upstart is looking for a motivated self-starter to manage our growing network of affiliate partners. These partnerships are a key part of our strategy to rapidly scale our user acquisition efforts as we become one of the largest marketplace lenders. The ideal candidate will have9/1/2015 Palo Alto, CA Save This Job
Job Locations US More information about this job: Overview: Join the frontlines. We?re always looking for exceptional product marketing, events management, and communications professionals to drive innovative campaigns, events, and programs. This is your chance to fearlessly8/18/2015 Redwood City, CA Save This Job
Job Type: Full Time Location: Belmont, CA Department: Marketing, Demand Generation The RingCentral environment is fast paced, high octane, success driven, team oriented and committed to growing the business in a virtually untapped market. Do you have the ability to thrive in8/17/2015 Belmont, CA Save This Job
The candidate will drive the execution of an integrated digital experience of A10 Web properties and portals with a global mindset. The site should be structured to maximize prospect and customer engagement, demand generation and conversions of inbound lead generation. This role8/17/2015 San Jose, CA Save This Job
As Hadoop adoption continues, newer Hadoop related open source technologies will continue to hit the market. Ability to spot trends, anticipate customer needs and innovate in an open source environment are critical to the success of Hadoop. In addition, the ability to stitch8/15/2015 San Jose, CA Save This Job
at Houzz Palo Alto Houzz is looking for a smart, top-notch writer and researcher to join our Palo Alto, CA team. You should be very web savvy, with a good understanding of how search engines work. You have experience in research, comparative analysis and content writing; you are8/9/2015 Palo Alto, CA Save This Job
at Houzz Palo Alto Houzz is looking for a Marketing Professional to assist with data collection and analysis. Flexible hours Great for college students to gain experience in the tech industry8/9/2015 Palo Alto, CA Save This Job
Responsibilites : Analyze technical (computer/software) press to identify target reporters Create database of reporters Write and distribute press releases Follow up on selected reporters (email and phone) Compensation: Per project and by results.9/7/2015 San Jose, CA Save This Job
- Qualifications: o Experiences with system level design methodology, HLS or VP applications o Experiences and knowledge of C++ and SystemC o Knowledge of OSCI TLM-2.0 standard o Experiences in model validation or RTL verification8/25/2015 Santa Clara, CA Save This Job
We are looking for motivated account managers who love driving successful outcomes for customers. You will partner with the customer to clearly communicate and realize meaningful results while working directly with engineering, product, and marketing.9/4/2015 Mountain View, CA Save This Job
We are looking for a Web Designer/Developer to join our growing team. This position will support our worldwide marketing efforts, taking control of the creation and maintenance of Silver Peak's digital presence as we continue our growth phase and ex10/8/2015 Santa Clara, CA Save This Job
The Technical Marketing Engineer, Cloud and Virtualization (TME) works with customers, sales, product management and engineering to drive successful introduction and adoption of Silver Peak's award-winning technology into private, public and hybrid9/19/2015 Santa Clara, CA Save This Job
Quantcast seeks a talented Software Engineer to join the Enterprise Applications team. We work directly with numerous business teams including Marketing, Sales, Planning, Client Services, and Finance to build awesome technical solutions supporting the Lea9/10/2015 San Francisco, CA Save This Job
Quantcast seeks a talented Senior Software Engineer to join the growing Enterprise Applications team. We empower our employees and business teams with critical business applications in areas such as Marketing, Sales and Order Management. The mission8/23/2015 San Francisco, CA Save This Job
Quantcast seeks a talented Senior Business Systems Analyst to join the growing Enterprise Applications team. We provide business teams with critical business applications in areas such as Marketing, Sales, Order Management and Finance.8/16/2015 San Francisco, CA Save This Job
Company Overview:headquartered in Mountain View, CA, is a world-class social analytics platform, chosen by world-famous brands such as Coca Cola, Ralph Lauren, Universal Music Group, Yum Foods and Nickelodeon to deliver social, live analytics for action - through industry leading9/17/2015 Mountain View, CA Save This Job
BloomReach Organic Search (BROS) is changing how products get discovered on the web. Leveraging BloomReach's core understanding of people, products, and web-wide demand, this solution optimizes our customers' websites to service demand for the millions of ways that their products8/6/2015 Mountain View, CA Save This Job
Location: Redwood City, CA Type: Full-time Time frame: Immediate About QuanticMind QuanticMind, the Platform for Smarter Advertising, is a rapidly growing software-as-a-service company. It is the performance leader in predictive advertising management software for paid9/21/2015 Redwood City, CA Save This Job
Marketing San Jose, CA The Senior Manager, Demand Generation and Global Campaigns will be a key member of the Demand Generation and Marketing Operations team, tasked with developing and leading the execution of end-to-end digital marketing campaigns supporting A10?s revenue9/16/2015 San Jose, CA Save This Job
Zenfolio, part of the Art.com family, is the leading platform used by more than 100,000 pro and enthusiast photographers to present and sell photography online. With the most complete set of features for professionals, Zenfolio helps photographers turn their passion into a8/31/2015 Menlo Park, CA Save This Job


text:
Live the beachfront condo dream in the Dominican Republic!
This two bedroom condo is only a few minutes away from the quaint and famous surf town Cabarete. If you are enthusiastic about learning how to kite board or windsurf you find wonderful schools along the beach in Cabarete. You can set up your kite on the lawn in front of your condo and get ready for a downwinder to kite beach (the kiter's hang-out) or Encuentro – the surfers' beach.
The condominium rents for USD 900.00 per month in high season and USD 700.00 per month in low season plus electricity. It includes the cooking gas, 24 hour electricity from the generator, security, cable TV, 32" flat screen, basic internet 512 kps by cable (can be upgraded) with backup internet by phone line.
See a virtual tour of the project here.


text:
Lithia back on track
|| Print |
|High Five
|Wednesday, April 28, 2010
Lithia Motors' revenue in the first quarter of 2010 reached $463 million, up 13.4%.
Rising demand for new vehicles was a key contributor to the Medford company's growth, with same-store new vehicle sales increasing 11.5% compared to the same period last year.
Read the full story at the Mail Tribune.
{biztweet}Lithia Motors revenue{/biztweet}
|The List: 100 Best Nonprofits to Work For in Oregon
|Run, Nick, Run
|One Tough Mayor
|100 Best Nonprofits: Working for equality inside and out
|Salad Days
|Cream of the Crop
|Keep Pendleton Weird
|2 out of 5 millennials pay for their news
|Oregon's graying workforce
|How much did Bernie Sanders raise in Q3?
|Federal regulators OK Jordan Cove LNG terminal
|Amazon to emulate parts of Uber's model
|Another former Daimler alleges discrimination
|Struggling Whole Foods announces layoffs
Wage gaps and workforce shortages are threatening the quality of care and supports to Oregonians with intellectual and developmental disabilities. Who's caring for those who care for our most vulnerable residents?
Engaging employees and customers along the way.
After first visiting as tourists, entrepreneurs relocate to Oregon and spur economic growth.
Are you planning a meeting, party, gala, fundraiser, holiday party, golf tournament, retirement party, team building or birthday? You won't want to miss this show to get hundreds of great ideas!
Promoting from within its own ranks, PacificSource Health Plans has tapped Tony Kopki to head its commercial lines of business in Oregon, Idaho and Montana. In his new role as Vice President of Commercial Programs, Kopki will provide strategic, product and market leadership for PacificSource's commercial programs.
Thomson brings 25 years of healthcare experience in provider relations, sales, marketing and communications.


text:
Obituary & Guest Book Preview for Rollo Eli Barrett
Rollo Eli Barrett Obituary
Published in Idaho Statesman on Jan. 20, 2013
First 25 of 285 words
: Rollo Eli Barrett Rollo Eli Barrett, 90 of Boise passed away peacefully with his family by his side on Monday January 14, 2013. Dad was...
View Full Obituary & Guest Book for Rollo Barrett
Rollo Barrett Guest Book | View 2 of 4 Entries:
View Additional entries from:
Previous 10
Next 10
View Complete List
View Full Obituary & Guest Book for Rollo Barrett
Ways to View
Restore Obituary & Guest Book
Keep the Obituary and Guest Book online for one of these time periods:
Permanent Sponsorship
$ 79.99
One-Year Sponsorship
$ 29.99
24-Hour Restoration
$ 2.95
Print Obituary & Guest Book
Order a professionally printed copy, starting at $39.00:
Deluxe Hardcover Book
$ 79.00
Laminated Hardcover Book
$ 59.00
Laminated Softcover Book
$ 39.00
Didn't find the Obituary you were looking for?
Return to today's Local Obituaries for the Idahostatesman.com.
Powered By
Search Obituaries
How To
Questions? Contact Us
Privacy Policy
© 2015 Legacy.com All Rights Reserved


text:
…picture, James reminds us what truly matters: simply the gospel. He challenges us to be "doers of the Word, and not hearers only." The gospel should define how we think, feel, and act. 40 Days to the Cross: Reflections from Great Thinkers Editors: Rebecca Van Noord and Jessi Strong … more
Just as God's promise of an eternal home raises joy and anticipation, so should our family relationships on earth. This guide offers practical help on building a strong spiritual family that is a comfort and support to each member's walk with God. Title: The Spirit-Filled Family … more$5.95
MacArthur Bible Studies: 1 & 2 Peter by John F. MacArthur
…to stand strong in the face of troubles and trials. Their study is crucial to the Christian walk. Title: 1 & 2 Peter Author: John MacArthur Series: MacArthur Bible Studies Publisher: Thomas Nelson Publication Date: 2000 more$6.95
…one minor Old Testament man whose godly line still continues today? The answer communicates the strong and sure message of hope in The Promise of Jonadab. Logos Bible Software dramatically improves the value of any resource by enabling you to find what you are looking for instantly and … more$9.95
Fasting: A Biblical Historical Study by Romara Dean Chatham
Many Christians believe that fasting promises power, strong faith, and health. In fact, there's a catalog of supposed spiritual, mental, and physical benefits. Fasting is presented as a sort of cure-all. But are these beliefs valid? Are they even biblical? This is the book that … more$6.99Reg.:
$11.95Save: $4.96 (42%)
2 Timothy: Finish-Line Faith by Matt Proctor
…rely on God's strengthening grace to finish strong. Product Details Title: 2 Timothy: Finish-Line Faith Author: Matt Proctor Publisher: 2009 Series: 3:16 Bible Commentary Series Publication Date: 2009 Pages: 112 About the Author Matt Proctor has served as a youth minister, preaching … more$7.95
Unquenchable Faith by David Faust
…tough place for new followers of Christ to grow up in their faith. Have you ever thought of the Apostle Paul as a concerned parent? Paul's letters to the church in Thessalonica are full of parental affection and advice. Once can't help but be touched by his strong desire to be with his … more$7.95
A Romans 12 Disciple Series (4 vols.) by Jim Grassi
…efficient and comprehensive research tools are in one place, so you get the most out of your study. Building a Ministry of Spiritual Mentoring by Jim Grassi The Spiritual Mentor Finishing Well, Finishing Strong More Than a Fisherman Building a Ministry of Spiritual Mentoring Series: A Romans … more$35.99Reg.:
$43.96Save: $7.97 (18%)
…Karlfried Froehlich's Sensing the Scriptures offers a valuable alternative. This clearly written and compelling book makes a strong argument for a new look at the centuries-old view of the multiple senses of Scripture, especially the four levels symbolized by Aminadab's chariot (Song of Songs … more$17.50Reg.:
$20.00Save: $2.50 (12%)
Bible Personalities: A Treasury of Insights for Personal Growth and Ministry by Warren W. Wiersbe
A good biographical example can captivate listeners and impart a strong message. But many times these effective examples are hard to find, requiring in-depth research and more time than a busy pastor or teacher can spare. Internationally known writer and former pastor Warren W. more$16.00
Ruth A. Tucker People of the Bible Collection (2 vols.) by Ruth A. Tucker
…from Their Surprising Stories Author: Ruth A. Tucker Publisher: Baker Publication Date: 2014 Pages: 336 Sample Pages: 1 | 2 | 3 | 4 | 5 | 6 | 7 Tour the lives of the Bible's most fascinating women with bible scholar Ruth A. Tucker. Edgy, strong willed, and controversial—the women of the … more$27.95Reg.:
$38.38Save: $10.43 (27%)
…Thanksgiving Last Words Satan's Success Conclusion Praise for the Print Edition The expositions are clear, vigorous, and strongly evangelical. There is little to which the critic can take exceptional there is much, very much, to edify and instruct the candid reader. We are very glad to give … more$41.95
…collection also discusses whether Jesus' followers in early church history worshipped him as a prophet or God. Religious and historical convictions strongly affect the many perspectives people have on Jesus and Christianity. By introducing thoughtful insight and contrasting ideas in relation … more$59.95
…fundamentals of Christianity to provide the reader with a strong foundation Includes work from dedicated biblical scholars Contains volumes meant to encourage the reader in their faith Product Details Title: What the Bible Says Series Publisher: College Press Volumes: 8 Pages: 3,033 Christian … more$185.95
…commentary of 1 and 2 Corinthians. It's a perfect reference and supplement for Sunday school teachers, eager students, and for pastors preparing sermons. In addition to a strong emphasis on edification of the church through spiritual gifts, Paul's letters to the church at Corinth deal directly … more$59.95


text:
Hi all,
I have tried to add a list of pages under the category menu bar which I have been able to do successfully, what I haven't been able to do is get it to display pages that are listed as 'child' under the main page.
I have added to the 'depth' field, but this hasn't helped.
How can I get the tab to display child pages under the main page listed?
This is what I have:

<li <?php wp_list_pages('title_li=&depth=3&sort_column=menu_order&include=3093,1089,988,67,2,38');?>
The pages are the correct ones and the 3093 page 'should' list sub pages but I do not know how to tell the code to list pages under the 3093 page...
I hope I have explained this correctly!
Thanks in advance for any help.


text:
All hotels in Port Charlotte, Florida, United States of America
1 room, 2 adults
24480 Sandhill Blvd, Port Charlotte, FL, 33983, United States of America, 855-238-1591
Hampton Inn Port Charlotte, Port Charlotte
Lowest price$99
available on 10/11/15
Best Price Guarantee
Port Charlotte hotel with outdoor pool, bar/lounge
- Free hot/cold buffet breakfast and free parking
Main amenities
- 71 smoke-free guestrooms
- Outdoor pool
- Bar/lounge
- Fitness center
- Business center
What's around
- Deep Creek Golf Club (2.6 mi / 4.2 km)
- Lake Betty Park (3.9 mi / 6.3 km)
- Deep Creek Preserve (4.4 mi / 7.1 km)
Enjoy your visit to Port Charlotte with a stay at Hampton Inn Port Charlotte. You can visit the fitness center for a workout or unwind with a drink at the bar/lounge.
We would stay at this hotel again. Rooms were clean and the beds comfortable. Nancy and…
ALL hotel staff are excellent and friendly .
TripAdvisor Traveller Rating: 4
From 274 reviews
Enter dates to check prices
Hotel Info: 866-539-8117from $99
- One king bed non smoking
- Room, 2 Queen Beds, Non Smoking, Balcony
- One King Bed, Non-Smoking, Accessible
- Room, 1 King Bed, Accessible, Non Smoking
- Room, 1 King Bed, Balcony, View
- Two Queen Beds, Smoking
- One King Bed, Smoking
- Two queen beds non smoking
Also popular with travelers from the USA
At a glance
Key facts
Hotel size
- This hotel has 71 rooms
- This hotel is arranged over 3 floors
- Check-in time starts at 3 PM
- Check-out time is 11 AM
Required at check in
- Credit card or cash deposit required
- Government-issued photo ID required
- Free self parking
Other information
- Smoke-free property
Nearby Two Star Port Charlotte hotels
In the hotel
Food and drink
- Free hot/cold buffet breakfast
- Bar/lounge
Things to do
- Outdoor pool
- Fitness facilities
Working away
- Business center
In the room
Home comforts
- Iron/ironing board
Be entertained
- Cable/satellite TV channels
Food and drink
- Microwave
Top Port Charlotte hotels
Recently reviewed hotels in Port Charlotte
- Motel 6 VeniceFrom
- Island Sun InnFrom
"The room was exceptionally clean, comfortable and quiet. The bathroom was a little tight. Shower older and water difficult to regulate. The pool area is very nice and there are…"Jul 8, 2015
- Inn & Suites of EnglewoodFrom
- Rodeway InnFrom
"Stayed here a couple times, convenient location and for the price much nicer then some other places i have stayed at. I will stay again."Oct 18, 2014


text:
Published: Tuesday, October 14, 2008 at 12:30 a.m.
Last Modified: Monday, October 13, 2008 at 11:10 a.m.
Jerry and Dina Parrish of Jackson Creek announce the engagement of their daughter, Amanda Marie Parrish, to Zachary Lee Stanley, son of Eddie and Susie Stanley of Lexington.
The couple will be married Oct. 18, 2008, at Cid United Methodist Church by the Rev. Doug Rowe.
She is a 2006 graduate of Southwestern Randolph High School and attends Randolph Community College.
He is a 2005 graduate of South Davidson High School and a 2008 graduate of Davidson County Community College.
Reader comments posted to this article may be published in our print edition. All rights reserved. This copyrighted material may not be re-published without permission. Links are encouraged.
Comments are currently unavailable on this article


text:
Davina McCall is self-conscious on TV
Davina McCall gets body-conscious watching herself on TV.
The 46-year-old presenter - who fronts 'Got To Dance' on Sky1 - is known for her enviable physique, but admits she still suffers from the occasional insecurity.
She explained to OK! magazine: ''On 'Long Lost Family', they filmed me from behind and I look at my bum and go, 'Good God - it's just so wide!' But I've got to say a big thank you to Kim Kardashian because she's made the bottom a hallowed thing. I'm so happy that my big bottom doesn't look that big anymore.''
Davina ran, cycled and swam 500 miles from Edinburgh to London for Sport Relief in February and recently admitted she and husband Matthew Robertson were in a ''tough place'' in the lead up to her challenge.
She said previously: ''We had a really difficult six months. Matthew's dad was ill, I was doing the challenge and he'd hurt his shoulder.''
She added: ''When you're in your twenties and you have a tough six months then that's a break up. But when you're married it's nothing.
''You always have to think when you are in a relationship that if you have a tough six months then it's nothing in the grand scheme of things.''
More news
- Beauty queen among The Apprentice 2015 candidates
- Lord Alan Sugar: Simon Cowell should shave beard off
- MasterChef's Gregg Wallace wants more kids
- Jennifer Metcalfe feels 'pressure' to look hot
- NTAs 2016 introduce new categories
- Gemma Atkinson wants baby before she's 32
- Stephanie Davis to join I'm A Celebrity... Get Me Out Of Here!
- Charley Webb: Let Debbie Dingle go quietly


text:
New Vauxhall Adam
We currently have 175 new Vauxhall Adam cars available on Auto Trader. Search and compare the widest range of new cars, on Auto Trader.
From sourcing Vauxhall car dealerships and booking test drives to aiding your decision with our unique compare tool and real car owner reviews, Auto Trader finds the best new car deal for you.
Vauxhall Adam owner reviews
16th October '14
Fab car poor mpg
Just purchased my adam after having an A3 audi for the last 7 yrs. car is fab to drive , so easy to park and you have all the benefits of cruise control inter link system , hill assist …
Find a dealer near you
Enter your postcode below to find reputable dealers in your area.
New Vauxhall Adam cars for sale
|Showing 1 result
|New Vauxhall ADAM Hatchback The Vauxhall Adam is a premium supermini to rival the MINI and Audi A1. It's perfectly suited to fashionable urban dwellers thanks to its small size and attractive features.
|£10,699 - £16,735 Versions matched: 175
- First
- Previous
- 1
- Next
- Last


text:
Keith Urban and Nicole Kidman's Sweetest Moments
Their marriage got off to a rocky start in 2006, but today Keith Urban and Nicole Kidman are stronger than ever.
"We were thrown into his alcohol problem three months into the marriage, and that was big," Kidman told Parade in 2008. "We became the closest we could become, because we had to bare our souls. We did 10 years of marriage in three months."
PHOTOS: How Nicole's face has changed
Urban, who celebrates his 44th birthday Wednesday, credits his 44-year-old bride (and mother of his two children) with giving him a new sense of purpose.
PHOTOS: Stars and their soulmates
"As far as being a married man, it is the best thing I have ever done," Urban raved to UK's The Sunday Telegraph in 2007. "And it's not just about being married; to me, it is about being married to her, to that woman because she has just saved my life -- absolutely saved it."
Get more Us! Follow us on Twitter, Friend us on Facebook, Subscribe to Us Weekly
Most Watched on Wonderwall
Fashion Offenders
Fergie, Sarah Jessica Parker, and Heidi Klum all get an F on this week's Fashion Offenders.Watch Video »
Diva Double Date
Beyonce and Alicia Keys double date with their famous hubbies on today's Celebs Gone Social.Watch Video »
Where Are They?
See what the cast of "American Beauty" is up to today.Watch Video »
J.Lo's Mini Me
Jennifer Lopez shares an adorable photo of her look-alike daughter, plus more celebritots on social media on today's Celebs Gone Social.Watch Video »
First Pitch
Prince Harry, Jessica Alba and more stars who threw out the first pitch at baseball games.Watch Video »
Aussie Babe
Sofia Vergara poses in a revealing neon bikini while her "Modern Family" castmates hit the beach in Australia on Celebs Gone Social.Watch Video »


text: Mens White Italian Style Short Sleeved Paisley Shirt by Jenson Samuel Shirts £24.99$54.15€36.98$41.11 In stock £6.95$15.06€10.28$11.43 delivery to United States Receive this item 5-10 working days from dispatch International delivery available size size Small Medium LargeXL 2XL size Small Medium LargeXL 2XL size Small Medium LargeXL 2XL size Small Medium LargeXL 2XL quantity by Jenson Samuel Shirts Location: Bingley, GB read seller's profile see seller's complete range more items from this seller Brown Leather Apple Macbook Pro Air 13' Sl... £15.99$34.65€23.66$26.30 Cravat And Handkerchief Wedding Set £15.99$34.65€23.66$26.30 Orange Leopard Leather Apple Macbook Pro A... £15.99$34.65€23.66$26.30 Men's Slim Fit Shirt With Floral Design £34.99$75.82€51.78$57.56 Premium iPad Mini Case Orange Leopard Prin... £11.99$25.98€17.74$19.72 White And Blue Floral Cotton Pocket Square £9.99$21.64€14.78$16.43 view all you may also like PRODUCT DETAILS DELIVERY RETURNS ask seller a question save for later This stunning white paisley shirt is perfect for anyone looking to set themselves out from the crowss this summer. Avaliable in a range of sizes These very fashionable slim fit shirt come in a verity of styles that would be perfect for any formal or casual occasion. it comes in a range of sizes to fit every body style. It would make a perfect gift and a excellent addition to any mans wardrobe. Product Specifications 80% cotton 20% polyester Washing machine safe Contrast collar Double button collar Small Medium Large XL 2XL made from: 100% Cotton dimensions: weight 255g product code: 356636 This product is dispatched to you by Jenson Samuel Shirts. MAINLAND UK order now and receive this item on or before price Free Delivery Friday 9 October FREEFREEFREEFREE Express Next Day Tuesday 6 October £4.95$10.72€7.32$8.14 NON-MAINLAND UK estimated delivery price Highlands & Islands of Scotland 2-5 working daysfrom dispatch £1.75$3.79€2.58$2.87 Northern Ireland 2-5 working daysfrom dispatch £1.75$3.79€2.58$2.87 Other UK Islands 2-5 working daysfrom dispatch £3.95$8.55€5.84$6.49 INTERNATIONAL DELIVERY estimated delivery price Western Europe 2-5 working daysfrom dispatch £5.00$10.83€7.39$8.22 Rest of Europe 5-10 working daysfrom dispatch £5.00$10.83€7.39$8.22 USA & Canada 5-10 working daysfrom dispatch £6.95$15.06€10.28$11.43 Asia & Australasia 5-10 working daysfrom dispatch £7.25$15.71€10.72$11.92 Rest of the world 5-14 working daysfrom dispatch £7.25$15.71€10.72$11.92 You have 28 days, from receipt of cancellable goods, to notify the seller if you wish to cancel your order or exchange an item. Please note: goods that are personalised, bespoke or made-to-order to your specific requirements, perishable products and personal items sold with a hygiene seal (cosmetics, underwear) in instances where the seal is broken are non-refundable, unless faulty. Read more about our returns policy.Should you choose to return or exchange your order you will need to deliver the item(s) to the UK, where this seller is based. Jenson Samuel Shirts's return policy


text: Selena Gomez Gives Fan Concert Tickets by Hilton Hater at August 15, 2013 9:58 am. Comments This is pretty cool. Watch this video to learn more about Selena Gomez and how she gave a fan free tickets to an upcoming concert. Share Tweet Email Edit Delete Follow Selena Gomez? Facebook Disqus Star: Selena Gomez Related Videos: Selena Gomez Videos Related Post: Selena Gomez Gifts Fan with Free Concert Tickets Uploaded by: Hilton Hater Uploaded: August 15, 2013 Duration: 71 seconds


text:
A new exhibition has just opened at the Stanford Music Library entitled Treasures from the Archive of Recorded Sound, on show through August 14. The exhibition was curated and installed by the Archive of Recorded Sound's Interim Operations Manager, Benjamin Bates, who describes the content and theme of the exhibit in more detail.
A manuscript fair copy of the text an otherwise unrecorded work, Agamemnon, opéra des dames, Poëme heroique, don't les paroles et la musique ont fait l'amusement d'un Particulier Vieux Stil, was recently acquired. It is believed to have originated in the second half of the eighteenth century and was apparently performed in Paris, "La Feste est au bord de la Seine près des Thuilleries" to mark the end of recent hostilities. The manuscript which also contains the text to three cantatas, L'innocence, L'amour vainquer, and Les regrets, originally scored for musette, vielle, flute, and bassoon, will be kept in the Department of Special Collections, Green Library.
2013 marked the 200th birthdays of Giuseppe Verdi and Richard Wagner. The Music Library purchased a number of out-of-print and rare scores of works by both composers, and books by and about Wagner. The Wagner works include his Die Kunst und die Revolution (Leipzig, 1849) and Deutscher Kunst und deutsche Politik (Leipzig, 1868), Julius Lang's Zur Versöhnung des Judenthums mit Richard Wagner (Berlin, 1869), and Hermann Schneider's Richard Wagner und das germanische Altertum (Tübingen, 1939). Arrangements for piano, 2-hands including overtures to Christoph Columbus (Leipzig, 1908), Faust (Leipzig, 189?), and König Enzio (Leipzig & London, 1908), and a score of Lohengrin in French (Paris 1891) were also acquired. All of the Wagner materials are described in Significant Acquisitions 2012-2013, on the Music Library's web page.
Stanford music librarians Jerry McBride, Mimi Tashiro, Nancy Lorimer, Jon Manton, Jihong Zhang, and Ray Heigemeir (that's me) were in attendance at the 83rd annual meeting of the Music Library Association in Atlanta, Georgia at the end of February. A total of 403 attendees from the US, Canada and select other countries spent the better part of a week learning, discussing, debating, and celebrating the changing landscape of music librarianship.
Stanford composer Brian Ferneyhough is the subject of the first monograph from Intellect Books' Critical Guides to Contemporary Composers series. Brian Ferneyhough, by Lois Fitch, "examines the critical issues fundamental to understanding the composer as both musician and thinker"--[book cover]
Chapters include: biography; notation; the solo works: 'Black Scherzo'; chamber 'concerts'; chamber music; string quartet; Time and Motion Study Cycle; Carceri d'Invenzione: style and innovation; Shadowtime; works for orchestra and large ensemble; and aesthetics.
Recently acquired, an engraving by Jan Sadeler (1550-1600) from 1590, based on the painting of Joos van Winghe (1544-1603) depicting King David playing the harp. A group of choristers is gathered around an open choir book which contains the 5-part setting of Psalm 116 by Andreas Pevernage (1542 or 3-1591).
A full-length woodcut portrait of the composer Coclico at age 52, printed in 1552, was recently purchased. It includes music notation at upper left, with the text "Desperando spero," and identification of the composer, "Adrian Petit Coclico Mvsico. Etat: LII," at upper right. This inscription is the only known source that states his birth year.


text:
This property is no longer supported by Rent.com. Pricing and description information may not be up-to-date. Check out other great properties in the area, we're sure we can find you the perfect home.
Ridgeview apartment offers spacious 2 bedrooms 2-bath apartments located in a quiet community of Vista, CA. This beautiful complex sits on a hillside giving a great view from apartment balcony. This unit has a ceiling fan, refrigerator, dishwasher and range oven.
There is an on-site laundry facility and common areas. We have an excellent on site manager and residents. There are also reserved parking spaces for your convenience. We do accept Housing Assistance vouchers. We are pet friendly with an additional deposit.
6, 9, and 12 month leases offered. Security deposit: $700. Pet deposit: $400 (refundable). 20lb weight limit on pets, max of 1 pet per unit.
Dogs Allowed. Cats Allowed. Call for service animal policy.
20 units
Don't have a Rent.com account? Create one.
Creating an account allows you to acccess your saved and contacted properties on any device. Already have an account?
Log In
We are experiencing technical difficulties at the moment. Please try again later.


text:
New Poll Shows Most Americans Believe LGBT Couples' Right to Marry is Protected by Constitution
June 6, 2014 by HRC staff
A new poll released by the Washington Post and ABC News shows that 50 percent of Americans believe that gay and lesbian couples have a constitutional right to marry guaranteed by the U.S. Constitution's Equal Protection clause. Additionally, 56 percent of Americans and 77 percent of those under the age of thirty support marriage rights for same-sex couples. Today's results are the latest in an ever-expanding trend showing Americans moving inexorably in the direction of supporting equality for same-sex couples.
In addition to 77 percent of 18- to 29-year-olds, marriage equality enjoys broad support from 30- to 39-year-olds -- 68 percent. Even 50 percent of those between the ages of 40 and 64 support marriage equality. During the 2012 presidential election, 84 percent of voters fell into one of these age brackets where there is majority support for marriage equality.
Among those who say they strongly oppose marriage equality, nearly half say it's not even "somewhat" important to them. Conversely, only 19 percent of strong marriage equality supporters put such low priority on the issue. In fact, 81 percent of strong supporters say it's at least "somewhat" important.
Other recent polls show rising support for marriage equality among various demographic groups. A March 2014 poll shows 61 percent of Republicans and Republican leaners under the age of thirty support marriage rights for gay and lesbian couples. One month earlier, the New York Times and CBS News released polling that puts marriage equality support among Catholics at 62 percent. More recent polling info is available online here.
Today 44 percent of Americans live in a jurisdiction with marriage equality. Same-sex couples can legally marry in nineteen states and the District of Columbia, while 31 states have a law or constitutional amendment restricting marriage to the union of one man and one woman.
For more information on the current marriage equality landscape and court cases in America, visit www.americansformarriageequality.org.
Top Posts
Time to Celebrate: National LGBT Pride Month
June 1, 2015
Frequently Asked Questions about the Upcoming Marriage Equality Case at the U.S. Supreme Court
April 24, 2015
Be Ready to Rally on Decision Day
June 15, 2015
What's at Stake: Potential for Nationwide Marriage Equality Following Supreme Court Ruling
June 5, 2015
Watch: HRC Answers Your SCOTUS Questions
June 19, 2015


text:
Last month, San Francisco mayor Gavin Newsom issued a proclamation declaring this summer, through August 2010, as another Soda Free Summer. Newsom's action was meant to encourage all San Franciscans to opt for healthy alternatives to calorically sweetened beverages.
In an effort to lead by example, Newsome has now issued an executive order that bans vending machines on city property from dispensing Coke, Pepsi and other calorically sweetened beverages. Sports drinks and artificially sweetened water also are included in the ban (MLive.com).
The California Center for Public Health Advocacy recently presented evidence demonstrating a direct link between soda consumption and obesity. Other studies have shown that sweetened beverage consumption trends parallel the rise in obesity.
The city of San Francisco has been on a crusade to address the public health crisis of obesity, inactivity, and poor nutrition, as well as the excessive waste produced by consumption of bottled beverages.
In June 2007, Mayor Newsom signed an executive order that banned the city from using taxpayer money to buy bottled water for its employees.
Despite owning a pristine mountain reservoir said to produce some of the country's best-tasting tap water, the city was estimated to spend nearly $500,000 a year on bottled water at the time the order was signed (SFGate).
MLive.com reports that "beyond the vending machine ban, San Francisco's Newsom also wants to impose a retail fee on soda."
Read more: coke, obesity, pepsi, real food, soda, soda ban, soft drinks
Image Credit: Flickr - dcJohn
banning just makes people want it more...
banning just makes people want it more...
Banning things works so well. Just ask the alcohol industry. We banned them and we got Al Capone and Joe Kennedy in return. Now if you want to tax the crap out of it, I could get behind that. Of course if you wipe out the soft drink industry, are you going to put all of the people who lose their jobs on the government payroll?
Banning things works so well. Just ask the alcohol industry. We banned them and we got Al Capone and Joe Kennedy in return. Now if you want to tax the crap out of it, I could get behind that. Of course if you wipe out the soft drink industry, are you going to put all of the people who lose their jobs on the government payroll?
Bravo sindaco.
I like this ban
Eminent American Arthur Rudolph http://en.wikipedia.org/wiki/Arthur_Rudolph
Eminent American Werner Von Braun http://en.wikipedia.org/wiki/Wernher_von_Braun
these Americans rose to national prominence and were highly acclaimed and decorated. And they were nazi war criminals.
maybe adding steep fees is the way to go, rather than banning altogether
thanks and noted
I don't drink soft drinks but I think that banning them is an infringment on my right to choose, even if it is a bad choice.
Disclaimer: The views expressed above are solely those of the author and may
not reflect those of
Care2, Inc., its employees or advertisers.
Food Security "101", a Thanksgiving meal grown and raised on ones property. The gardens bounty stored…
Sorry about the typos I always see too late. Oh, for an "edit" button!
I would be very afraid for the dog. I would tell him not to do it again!
Problem on this page? Briefly let us know what isn't working for you and we'll try to make it right!


text:
Subscribe to our newsletter for saving tips and our favorite deals of the week.
Water Warehouse carries water pumps, heaters, filters, chlorinators, and many more supplies. All of the supplies are offered for at least 30% off. Water Warehouse is a direct shipper, so customers are assured of speedy delivery. There are no middlemen.


text:
'Twin' paths diverge now study starts
Different plans for tertiary studiesJAMES GREENLAND
Relevant offers
Close friends Michaela Matenga and Ella Marwick "are like twins", but this year will follow different paths to pursue tertiary studies at opposite ends of the island.
The young women played rep netball and touch rugby together in Nelson, but attended different secondary schools.
Both start tertiary study this year and hope for a career in health services, but Michaela is heading south to Otago and Ella will remain in Nelson.
Michaela, 18, was Nayland College's top female athlete in 2012, and Ella, 17, won the equivalent "general sporting excellence" award at Waimea. Both have featured in the Nelson Mail for their sporting successes, but 2013 will bring them a new academic challenge.
And for Michaela, who headed south for Otago University today, there is the added challenge of starting a new life in Dunedin.
The first in her "huge" family to attend university, she plans to complete a foundation course at Otago to prepare her for the gruelling first-year health-science course, which determines who is accepted into the university's highly competitive medical school.
She was accepted into Arana hall of residence, a popular boarding hostel that prides itself on hosting leading students from around the country.
Michaela will be assisted in her studies by two scholarships; Tu Kahika, available to about 20 Maori health-science students each year, and a Maori and Pacific Island students Otago University entrance scholarship. Each scholarship would contribute about $10,000 toward Michaela's expensive eight-year degree, which could easily leave a student under a mountain of debt, she said.
"Obviously the money is nice," she said. "But the main reason it is good is there is heaps of support. It's not like you are going down to uni by yourself."
Ella is staying in Nelson, studying nursing at the Nelson Marlborough Institute of Technology on their half-a-degree for free scheme, which she said was about $20,000 cheaper than studying away from home.
Her course began on Monday and already Ella can see that she has chosen the right path.
"I have always wanted a job where you feel like you are putting your energy into something that's worthwhile.
"Once you start you kind of know, or you don't, and I know it's the right path."
She said an accountant's salary was attractive, but that kind of work would bore her. Instead, she would eventually specialise her training, possibly to become a paramedic. "I want a bit of excitement in my life," she said.
Her initial training would take three years and involve practical placements at hospitals and medical centres around the region.
Michaela's Nayland College science and maths teachers inspired her health-science ambitions during recent years, she said, while her Porirua upbringing instilled a desire to help others.
"It wasn't the best growing up, surrounded by violence and that. So, I have always wanted to help people. Help them come out if it, I suppose," she said.
"I am not only doing it because I enjoy the physical, fixing side, but I also want to talk to people too."
Michaela will miss her family in Nelson and Ella too, but will visit whenever she can.
- The Nelson Mail
Sponsored links
Hot air boosts Nelson Red Peak campaign
Prison van forced to turn back
Politics and social justice run through O'Connor sisters' blood
Nelson boy gets Rugby World Cup call up
Tasman District Council supports higher-density housing in Richmond
Nelson council planning 'expedient' action on Brook reserve, group says
Pooky the lamb dies twice and survives
Choirs come together in Mass for Peace
New QYouth coordinator keen to make Nelson a safer space
Nelson police crack down on boy racers
Restaurateurs gather in Nelson for hospitality conference
Waimea Dam will benefit apple growers, says Primary Industries Minister
Man seriously assaulted before keys stolen in road rage attack near Nelson
Nelson boy gets Rugby World Cup call up
Hot air boosts Nelson Red Peak campaign
Tasman District Council supports higher-density housing in Richmond
Politics and social justice run through O'Connor sisters' blood
Masked Parade brings book characters to life
Letter fuels grey battle for power
Thai flavours hit Nelson waterfront
Parents of missing Dutch tourist keep searching
Couple lucky to escape Stoke house fire
Craft brewing capital title under threat
Incentives to boost Nelson's inner-city living
Embattled Grey Power president withdraws from fight
Politics and social justice run through O'Connor sisters' blood
Prison van forced to turn back
Nelson boy gets Rugby World Cup call up
Pooky the lamb dies twice and survives
Hot air boosts Nelson Red Peak campaign
Nelson mountainbike riders make their mark on the world stage
Power pole on top of car in Motueka
Controversial blue cod rules ditched
Tasman District Council supports higher-density housing in Richmond
Brave production offers honesty and hope
Nelson council planning 'expedient' action on Brook reserve, group says
Do you agree with the city council's decision to put a 30-minute limit on buskers' performances?Related story: (See story)


text:
Appointments & Offices
Check insurance plans, locations and make an appointment
Office Locations
- Plastic Surgery Assocs Tulsa
- 6585 S Yale Ave Ste 1020
- Tulsa, OK 74136
- (918) 481-2900 (Office)
|Hospital Quality
|Patient Satisfaction
|Appointments & Offices
© Copyright 2015 Healthgrades Operating Company, Inc. Patent US Nos. 7,752,060 and 8,719,052. All Rights Reserved.
Third Party materials included herein protected under copyright law.
Use of this website and any information contained herein is governed by the Healthgrades User Agreement.


text: We were greeted with a friendly welcome.....our room was very comfortable with a shower and views to die for. We loved the rural setting and the very friendly animals even the sheep greeted us on our return in the evening :) Everyday we were provided with a huge breakfast along with scrumptious home made scones. Vicky was always available and... More


text:
The German is the hot favourite to replace Brendan Rodgers at Anfield and Redknapp believes he would be a big success
Manchester United's summer signing is enjoying a superb start to life in England, scoring four goals in six games since joining from Monaco
The Reds boss had previously claimed that a media "frenzy" had been whipped up that was aimed at getting him the chop at Anfield
The Sky Sports pundit has been impressed with the former Chelsea man's display in United's midfield so far this season
Liverpool lost 3-1 to bitter rivals Manchester United on Saturday, leaving them eight points off the top of the table after just five games
Jamie and Louise Redknapp hit the town with Boy George to celebrate his new judging role on The Voice
Freddie, 37, jokes that the rest of the boys were very disappointed that James Corden hadn't "gone all Hollywood" since he moved to host The Late Late Show in Los Angeles
With Gary Neville and Arsene Wenger trading blows, here are the finest ever feuds between pundits and managers
'If his little kid wants a dummy it's no problem with me,' insists former England star
The lead actor and his stunning Humans actress girlfriend had quite the star-studded support in London tonight
With the new Premier League season about to kick-off, here is a half century of things absolutely guaranteed to happen
A spokeswoman for the couple has ruled out recent reports connecting the couple with the ballroom contest
The famous couple may clash on the dancefloor after Jack Whitehall reportedly blurted out on the surprise news
Clyne will be the ninth player of the Premier League era to represent both Liverpool and Southampton. Can you name the other eight?
Tired of hearing television pundits talk rubbish? Now you can make them say whatever you want (sort of)
Wayne Rooney this week claimed the Red Devils were close to mounting a title challenge, but Redknapp reckons they've got it all to do next season
Neville and Redknapp give the full, touchscreen punditry treatment to some belting goals - scored by chubby blokes in parks across the land
The Frenchman lined up against former team-mates Pires and Campbell with his new colleagues, as the two sides exchanged 10 goals
Sky Sports' handsome man talks to Row Zed readers - about beating a team of Nevilles, his legendary upside down pineapple cake and sellotaping a bulldog to a lamppost
Rappers Delight has never sounded so bad, erm... we mean good
The four players were all linked with a move away from Old Trafford earlier in the season but each has become a pivotal cog in Louis van Gaal's machine
Two of Sky Sports' finest pundits, in the Saturday Night Football studio, analysing "Tommo's" goal for "Dynamo Kebab" in great detail? Er, WHAT?
After the BBC scheduled the FA Cup semi-final clash between Arsenal and Reading to coincide with a Sky Sports Premier League fixture, we answer the question everyone's asking...
Jamie Redknapp shows us all exactly why he is a football pundit with his brilliantly pointless analysis of Manchester City
Jamie Redknapp suggested the Aussie midfielder could have "broken Sakho's jaw" and Crystal Palace chief Pardew is unhappy
Keys and Redknapp worked together on the North London derby draw at the Emirates back in 2008 as Spurs came from behind to snatch a point
The war of words between Jose Mourinho and Jamie Redknapp is threatening to spill over. If it does (here's hoping), we've figured out who might come out on top
The Special One once more claims there is a "campaign" against his men following their League Cup semi-final - but who precisely was he talking about?
The Special One once more claims there is a "campaign" against his men after TV analyst says striker deserved to be sent off over clash with Emre Can
With two little lads keeping them busy there was little opportunity for the couple to enjoy some alone time


text:
Tonight's show features Melanie Cogdill and Elliot Miller, the editors of the Christian Research Journal.This newest issue contains great articles about Tony Robbins,Utilitarianism, and MORE!
More Questions and Answers with Hank
Q&A: Kenneth Copeland, Age of Accountability, and Joseph Prince
Q&A: the Trinity, Antichrist, and Seventh Day Adventists
Best of BAM: Hath God Said? with Joe Dallas, and Q&A
Stealing from God – Part 1 with Frank Turek, and Q&A
Equipping the Next Generation, and Q&A
Those Who Teach Will be Judged More Strictly, and Q&A
Going Global – Part 2 with Larry Johnston, and Q&A
John Hagee, the Final Blood Moon, and Q&A
Best of BAM: Q&A: Shemitah, Forgiveness, and Speaking in Tongues
Debunking Date-Setters, and Q&A
Q&A: Jonathan Cahn, the Afterlife, and Jesus Calling
Are We on the Verge of Doomsday?, and Q&A
Hath God Said? with Joe Dallas, and Q&A
Discerning the Pope's Opinion on Climate Change, and Q&A
Best of BAM: Developing Discernment in Devotional Reading with Adam Pelser, and Q&A


text:
Josh joined NEA in September 2004 as a Venture Partner but has worked closely with NEA since his time as an NEA EIR back in 1995. He is currently a General Partner on NEA's healthcare team and focuses primarily on medical devices and consumer health technologies. Josh is also the Founder & Chief Executive Officer of ExploraMed I, II, III, and IV Inc., a series of NEA portfolio medical device incubators based on the West Coast that have created eight companies over the past 20 years. Acclarent, Inc, an ExploraMed II company, was acquired by Johnson & Johnson in 2009. EndoMatrix, Inc. was acquired by C.R. Bard in 1997 and TransVascular, Inc. was acquired by Medtronic in 2003 (both companies were created by ExploraMed I). On-going ExploraMed/NEA ventures include NeoTract, Inc. (created by ExploraMed II), Moximed, Inc. (created by ExploraMed III) and Nuelle, Inc and NC7 (both created by ExploraMed IV). In addition, Josh serves a part-time role on the faculty of the Stanford University Medical School as a Consulting Professor of Medicine and is Co-Founder of Stanford's Biodesign Innovation Program. Josh serves on the board of directors for DOTS Devices, ExploraMed II, III and IV, Intrinsic Therapeutics, Inc., NeoTract, Inc., Moximed, Inc., Nuelle, Inc, NC7 Inc. and Coravin. Prior to NEA, Josh was Founder and Manager of Pfizer's Strategic Innovation Group. Josh holds over one hundred patents for various medical devices in the fields of cardiology, ENT, general surgery, drug delivery, obesity, orthopedics, women's health, and urology. He received an MBA from Columbia University, a medical degree from the New York University School of Medicine, and a bachelor's degree in Mechanical Engineering from the Massachusetts Institute of Technology.
- August 26, 2015
- September 10, 2015
- June 11, 2015


text:
Jun 5, 2014; San Antonio, TX, USA; San Antonio Spurs forward Boris Diaw (33) drives to the basket during the second quarter against the Miami Heat in game one of the 2014 NBA Finals at AT&T Center. Mandatory Credit: Soobum Im-USA TODAY Sports
NHL 12m ago
This documentary perfectly describes the agony of being a Maple Leafs fan
It's been nearly 50 years since the Toronto Maple Leafs last won the Stanley Cup, and in that period the Leafs have never advanced past (…)
MLB 41m ago
8 consecutive reasons the Nats shouldn't hire Cal Ripken Jr.
Don't do it, Nationals.
MLB 42m ago
The 5 greatest photos from the Astros Wild Card celebration
The Houston Astros advanced to the American League Division Series on Tuesday with a win over the Yankees and like many clubs will and have (…)
Culture 45m ago
Matthew McConaughey destroys dreams of millions, is bald and overweight now
This is Matthew McConaughey. This is also Matthew McConaughey. But Matthew McConaughey doesn't look anything like those two Matthew (…)
NFL 46m ago
Youth football team has the saddest onside kick attempt ever
Oh no.
NFL 58m ago
Home belonging to Johnny Manziel's family destroyed in fire
A home belonging Johnny Manziel's grandfather Paul in Whitehouse, Texas, was destroyed in a fire early Wednesday morning. No (…)
NCAAF 1hr ago
Botched field goal leads to the most ridiculous two-point conversion ever
A brief synopsis of what happened during an extra point during Rowen's 14-9 DIII victory over Christopher Newport last weekend: (…)


text:
Search the Collection
Sitter in 1 portrait
by Alman & Co
Nathaniel Seymour Thomas(1867-1937), Bishop of Wyoming; son of Elisha Smith Thomas
Sitter in 1 portrait
by Alman & Co
vintage bromide print, circa 1910s
NPG x159536
Related People
- Elisha Smith Thomas (father)
Tell us moreback to top
Can you tell us more about this person? Spotted an error, something missing, or do you know anything that we don't know? If you have any information to share please complete the form below