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|"Aaron T. Myers (JIRA)" <j...@apache.org>
|[jira] [Commented] (HDFS-3980) NPE in HttpURLConnection.java while starting SecondaryNameNode.
|Wed, 17 Oct 2012 00:36:03 GMT
[ https://issues.apache.org/jira/browse/HDFS-3980?page=com.atlassian.jira.plugin.system.issuetabpanels:comment-tabpanel&focusedCommentId=13477495#comment-13477495 ] Aaron T. Myers commented on HDFS-3980: -------------------------------------- Hi Brahma, can you tell me exactly what you did to generate the keytab in question? It's unfortunately quite easy to accidentally invalidate a keytab for a given principal if you later export another keytab including entries for the same principal. The fact that servicePrincipal is null is interesting, and I would guess that it's due to having not properly configured the relevant principal setting in your configs. One things confuses me, however: I can't find the method KerberosUtil#getWebDfsPrincipalAndReset in any version of Hadoop that I'm aware of. What exact version are you experiencing this with? > NPE in HttpURLConnection.java while starting SecondaryNameNode. > --------------------------------------------------------------- > > Key: HDFS-3980 > URL: https://issues.apache.org/jira/browse/HDFS-3980 > Project: Hadoop HDFS > Issue Type: Bug > Components: security > Affects Versions: 3.0.0, 2.0.1-alpha > Reporter: Brahma Reddy Battula > Priority: Critical > Attachments: core-site.xml, hdfs-site.xml > > > Scenario: > ======== > I started secure cluster by going thru following.. > https://ccp.cloudera.com/display/CDHDOC/CDH3+Security+Guide.. > Here SecondaryNamenode is getting shutdown by throwing NPE.. > Please correct me If I am wrong... > Will attach conf and logs.. -- This message is automatically generated by JIRA. If you think it was sent incorrectly, please contact your JIRA administrators For more information on JIRA, see: http://www.atlassian.com/software/jira
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Tissue morphology and knowledge of the localization of specific proteins within a cell are integral to understanding cellular metabolism and the causes and consequences of diabetes and new advances for the treatment of diabetes. The tools of morphological analysis are varied but most involve particular skills or equipment that are too costly to be supported in individual laboratories. The objective of the Advanced Microscopy Core has been to make sophisficated morphological techniques readily available to all invesfigators ofthe Joslin DRC. The Core provides a wide range of services covering the broad categories: performing specific morphological procedures, providing access to equipment and reagents, and giving advice, training and interpretation. There are five main components in this core: Electron Microscopy (EM), Confocal, Histology and Laser Capture Dissecfion (LCM) facilifies and consultafion. Each of the components have specific operafional modes: the plastic embedding and electron microscopic work is done by the Core Manager, a senior highly experienced electron microscopist;the Core Histology Technicians take fixed/frozen fissue and returns both H&E and unstained slides for examinafion by the investigator. Both the Confocal microscopes and Laser Capture Dissecfion equipment are used direcfiy by the individual investigators with the Core providing training of new users, maintenance of the microscopes, and troubleshooting problems. The ultimate goal in all core activities is to provide prompt, customized, high-quality service. The core personnel are readily available for providing advice, troubleshooting and assistance with data interpretation to the users. The core provides every reasonable effort to help DRC investigators have access to a wide range of cutfing-edge morphological techniques.
|Pavkov, Meda E; Nelson, Robert G; Knowler, William C et al. (2015) Elevation of circulating TNF receptors 1 and 2 increases the risk of end-stage renal disease in American Indians with type 2 diabetes. Kidney Int 87:812-9
|Avadhani, Radhika; Fowler, Kristen; Barbato, Corinne et al. (2015) Glycemia and cognitive function in metabolic syndrome and coronary heart disease. Am J Med 128:46-55
|Baskaran, Charumathi; Volkening, Lisa K; Diaz, Monica et al. (2015) A decade of temporal trends in overweight/obesity in youth with type 1 diabetes after the Diabetes Control and Complications Trial. Pediatr Diabetes 16:263-70
|Rasbach, Lisa; Jenkins, Carolyn; Laffel, Lori (2015) An integrative review of self-efficacy measurement instruments in youth with type 1 diabetes. Diabetes Educ 41:43-58
|Castiglioni, Alessandra; Hettmer, Simone; Lynes, Matthew D et al. (2014) Isolation of progenitors that exhibit myogenic/osteogenic bipotency in vitro by fluorescence-activated cell sorting from human fetal muscle. Stem Cell Reports 2:92-106
|Markowitz, Jessica T; Volkening, Lisa K; Laffel, Lori M B (2014) Care utilization in a pediatric diabetes clinic: cancellations, parental attendance, and mental health appointments. J Pediatr 164:1384-9
|Isganaitis, Elvira; Woo, Melissa; Ma, Huijuan et al. (2014) Developmental programming by maternal insulin resistance: hyperinsulinemia, glucose intolerance, and dysregulated lipid metabolism in male offspring of insulin-resistant mice. Diabetes 63:688-700
|Teo, Adrian Kee Keong; Valdez, Ivan Achel; Dirice, Ercument et al. (2014) Comparable generation of activin-induced definitive endoderm via additive Wnt or BMP signaling in absence of serum. Stem Cell Reports 3:5-14
|Lee, Hyung-Yul; Wei, Dan; Loeken, Mary R (2014) Lack of metformin effect on mouse embryo AMPK activity: implications for metformin treatment during pregnancy. Diabetes Metab Res Rev 30:23-30
|Sanders, Kaitlyn; Jung, Jin Hyuk; Loeken, Mary R (2014) Use of a murine embryonic stem cell line that is sensitive to high glucose environment to model neural tube development in diabetic pregnancy. Birth Defects Res A Clin Mol Teratol 100:584-91
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Importance of spliceosomal RNP1 motif for intermolecular T-B cell spreading and tolerance restoration in lupus
- Fanny Monneaux1,
- Véronique Parietti1,
- Jean-Paul Briand1 and
- Sylviane Muller1Email author
DOI: 10.1186/ar2317
© Monneaux et al.; licensee BioMed Central Ltd. 2007
Received: 05 July 2007
Accepted: 26 October 2007
Published: 26 October 2007
We previously demonstrated the importance of the RNP1 motif-bearing region 131–151 of the U1-70K spliceosomal protein in the intramolecular T-B spreading that occurs in MRL/lpr lupus mice. Here, we analyze the involvement of RNP1 motif in the development and prevention of naturally-occurring intermolecular T-B cell diversification. We found that MRL/lpr peripheral blood lymphocytes proliferated in response to peptides containing or corresponding exactly to the RNP1 motif of spliceosomal U1-70K, U1-A and hnRNP-A2 proteins. We also demonstrated that rabbit antibodies to peptide 131–151 cross-reacted with U1-70K, U1-A and hnRNP-A2 RNP1-peptides. These antibodies recognized the U1-70K and U1-A proteins, and also U1-C and SmD1 proteins, which are devoid of RNP1 motif. Repeated administration of phosphorylated peptide P140 into MRL/lpr mice abolished T-cell response to several peptides from the U1-70K, U1-A and SmD1 proteins without affecting antibody and T-cell responses to foreign (viral) antigen in treated mice challenged with infectious virus. These results emphasized the importance of the dominant RNP1 region, which seems to be central in the activation cascade of B and T cells reacting with spliceosomal RNP1+ and RNP1- spliceosomal proteins. The tolerogenic peptide P140, which is recognized by lupus patients' CD4+ T cells and known to protect MRL/lpr mice, is able to thwart emergence of intermolecular T-cell spreading in treated animals.
Longitudinal studies of spontaneously lupus-prone inbred mouse strains and patients with systemic lupus erythematosus (SLE) consistently show an ordered appearance of typical auto-antibodies in the serum of individuals [1–4]. With time the fine specificity of the antibody response initially focused against one or few autoepitopes diversifies to other epitopes of the same protein (intramolecular spreading) and to other components that are physically associated within the same antigenic macromolecular particles, such as nucleosome, spliceosome, and Ro particle (intermolecular spreading). Epitope spreading is thus a process whereby epitopes distinct from and non-cross-reactive with an inducing epitope become major targets of an ongoing immune response. This phenomenon is not limited to autoimmunity; it has also been described in experimental and natural situations as a consequence of acute or persistent infection. Although the concept of epitope spreading was introduced more than 15 years ago [5], the cellular components that catalyze the spreading hierarchy have not been well defined, and certain aspects of this process remain unexplained. Recent studies suggest that autoreactive B cells are important cellular mediators contributing to autoreactive T-cell response diversification via their functions that mediate antigen processing and presentation [6, 7].
Previous work from our laboratory demonstrated that peptide 131–151 of the spliceosomal U1-70K protein as well as a peptide analogue containing a phosphoserine residue at position 140 (peptide P140) are recognized by CD4+ T cells from lupus mice. Both peptides were shown to behave as promiscuous epitopes and bind a large panel of murine and human MHC class II molecules [8–11]. Administration into young MRL/lpr lupus-prone mice of P140 peptide in saline, but not of the non-phosphorylated peptide 131–151, led to a dramatic amelioration of the clinical and biological manifestations of treated animals and significantly prolonged their survival [9]. The peptide P140 administrated in Freund's adjuvant (FA) accelerated the renal disease in MRL/lpr mice [9]. Our studies revealed further that peripheral CD4+ T cells from lupus patients, but not from patients with other autoimmune diseases (such as rheumatoid arthritis, primary Sjögren's syndrome, autoimmune deafness, polymyositis, primary billiary cirrhosis and autoimmune hepatitis) or infectious diseases, very specifically recognized the 131–151 and P140 peptides, and that phosphorylation of Ser140 prevented ex vivo proliferation of lupus patients' CD4+ T cells but not secretion of high levels of regulatory cytokines [11].
The 131–151 sequence of the spliceosomal U1-70K protein is located within a 80–90 amino acid-long RNA-binding domain. It encompasses a conserved sequence, called RNP1 motif, which is also present in other RNA-binding proteins, such as small nuclear (sn)RNP (such as U1-A) and heterogeneous nuclear (hn)RNP (such as hnRNP-A2/B1) proteins. Starting from the observation that sequences containing this RNP1 motif are often targeted by antibodies from lupus patients and mice, we hypothesized that the RNP1 motif could be involved in the earliest stages of the T-B intramolecular diversification process to other regions of one of the spliceosomal proteins that contain this unique motif, and might promote intermolecular spreading to epitopes of other proteins present within the same spliceosomal particle and containing or not an RNP1 motif [12, 13]. We demonstrated that an intramolecular T and B cell spreading effectively occurs in MRL/lpr mice tested at different ages and emphasized the importance of the RNP1 region in the cascade of events observed in the murine lupus response [14]. In the present study, we investigated the potential capacity of peptide 131–151 to generate antibodies that react with spliceosomal peptides and proteins encompassing, or not, the RNP1 motif. We tested the reactivity of peripheral blood lymphocytes (PBLs) from unprimed lupus mice to react with RNP1-containing peptides from several spliceosomal proteins. We also examined whether administration of protecting P140 peptide into MRL/lpr mice could modulate intermolecular epitope spreading in vivo by preventing T-cell response to peptides from heterologous spliceosomal proteins that do or do not contain an RNP1 motif. We provide data showing that P140 peptide treatment efficiently suppresses T-cell responses to other, unrelated spliceosomal epitopes, in addition to the tolerizing peptide. Remarkably, however, the B and T cell immune response to foreign antigens (such as viral particles) remains unaffected in treated animals.
Materials and methods
Peptides and proteins
Rabbit immunization
On days 1, 21 and 42, outbred Fauves de Bourgogne female rabbits (Dombes Romans, Chatillon, France) were injected subcutaneously (sc) with peptide 131–151 of the U1-70K protein (100 μg/rabbit/injection) emulsified in complete FA (CFA) for the first injection and incomplete FA for the subsequent injections. Rabbits were bled before immunization and then regularly throughout the protocol.
ELISA and Western immunoblotting
Standard ELISA procedures with peptide 131–151 (2 μM in 0.05 M carbonate buffer, pH 9.6) or dsDNA (100 ng/ml in 25 mM citrate buffer, pH 5.4) directly coated onto polyvinyl microtiter plates (Falcon, Oxnard, California, USA), and goat anti-rabbit IgG-horseradish peroxidase (HRP) conjugate (Jackson ImmunoResearch Laboratories, West Groves, Pennsylvania) diluted 1:50,000 in phosphate-buffered saline containing 0.05% Tween (PBS-T) were used to measure antibody reactivity in rabbit antisera [8]. The final reaction was visualized with H2O2 and 3,3',5,5'-tetramethyl benzidine used as chromogen, and absorbance was measured at 450 nm. For competitive ELISA, increasing amounts of inhibitor peptides were first incubated for 1 h at 37°C and then overnight at 4°C with constant dilutions of rabbit antisera in PBS-T. The mixtures were subsequently transferred to polyvinyl Falcon plates pre-coated with peptide 131–151, and post-coated with bovine serum albumin in PBS-T (0.4% w/v). After a 1 h incubation at 37°C and washing, bound antibodies were detected as indicated above. Preliminary experiments were conducted in a direct format to define the working dilution of rabbit antiserum. For Western immunoblotting, U1-A, U1-C, U1-70K, SmBB' and SmD1 proteins were first subjected to electrophoresis in 12.5% SDS-polyacrylamide gels and then transferred to nitrocellulose. The blotted strips were saturated in Tris-buffered saline, pH 7.5, containing 0.5% Tween (TBS-T) and 5% milk for 1 h at room temperature (RT), and then incubated with rabbit antisera diluted 1:500 in TBS-T-milk for 1 h at RT. After washing, strips were incubated with HRP-conjugated second antibodies to rabbit IgG (1:5,000 in TBS-T). Enhanced chemiluminescent (ECL™) reagents (Amersham Pharmacia Biotech, Buckinghamshire, UK) were used to reveal positive reactions.
Cellular assays
To study the natural T-cell reactivity occurring in unprimed lupus-prone mice, MRL/lpr mice were bled at weeks 8, 10, 12 and 14, and lymphocytes were purified by density separation (Lympholyte-M, d = 1.0875; Cedarlane, Hornby, Canada). PBLs were collected, washed three times, and resuspended at 3 × 106 cells/ml in L-alanyl-L-glutamine-enriched RPMI 1640 medium (Cambrex, Verviers, Belgium) containing 10% fetal calf serum (FCS; Dutscher, Brumath, France), HEPES, gentamycine, and β-mercaptoethanol. The proliferative response to peptides was measured in duplicate using 3 × 105 cells/well and a single peptide concentration (100 μM). After 72 h, the cultures were pulsed for 18 h with [3H]-thymidine (6.7 Ci/mmol; 1μCi/well) and DNA-incorporated radioactivity was measured using a Matrix 9600 direct beta counter (Packard, Meriden, Connecticut, USA; cpm range from 100 to 30,000). The SD of duplicate cultures was always below 20% of the mean. Control tests were performed by adding Con-A (100 μl/well; 5 μg/ml) to cells during the time of the culture (90 h).
To analyze the effect of P140 peptide administration on spontaneous T cell spreading occurring in lupus-prone mice, MRL/lpr mice received peptide P140 (100 μg in saline/mouse/injection; 10 mice) or PBS alone (10 mice) intravenously (iv) at weeks 5, 7 and 9 [9]. Treated mice were bled at week 10, and peptides of interest (100 μM in the cultures) were tested for their ability to induce proliferation of PBLs, as described above.
To examine the possible effect of P140 treatment on the ability of mice to mount a normal immune reaction after viral infection, P140-treated MRL/lpr mice (18-week-old at the time of the experiment) were challenged intranasally (in) with A/NT/60/68 influenza virus (H3N2 strain) in allantoic fluid using a dose predetermined to provoke ~30% of weight loss [19]. The required dose was of the order of 40 μl of undiluted allantoic fluid (HA titer 1260) per mouse. As a control, a group of untreated MRL/lpr mice was challenged in parallel. Mice were followed daily for their body weight. The capacity of PBLs from challenged mice to proliferate and secrete INF-γ ex vivo in the presence of increasing concentrations of the HA peptide 307–319 (PKYVKQNTLKLAT [20]) representing a promiscuous T-helper epitope from influenza virus HA, was evaluated 13 days after challenge, as described above. The anti-virus antibody production in the serum from challenged mice was measured in ELISA [19] 28 days after challenge.
All animal experiments were performed with the approval of the local Institutional Animal Care and Use Committee (CREMEAS).
Analysis for statistically significant differences was performed with Student's t-test. P values < 0.05 were considered significant.
Recognition by T cells from unprimed MRL/lpr mice of peptides containing or corresponding exactly to the RNP1 motif
In order to determine whether MRL/lpr T cells precisely recognized the RNP1 motif present in the respective spliceosomal proteins, we synthesized three additional peptides corresponding exactly to the RNP1 motifs of the U1-70K, U1-A and hnRNP-A2 proteins, called RNP1-70K, RNP1-U1A and RNP1-A2, respectively (Figure 1a), and tested, as above, the ability of these peptides to stimulate the proliferation of PBLs from 8-14-wk-old MRL/lpr mice (Figure 2b). No or very weak proliferation was observed in response to the RNP1-70K peptide 139–151 at any age. A modest but significant proliferative response with RNP1-A2 peptide 45–57 was measurable with PBLs from 10, 12 and 14-wk-old MRL/lpr mice. In contrast, as early as 8 weeks of age, and at least until 14 weeks, a strong proliferative response was observed when MRL/lpr PBLs were cultured in the presence of the 13-residue-long RNP1-U1A peptide 47–59 (Figure 2b; maximum at week 12). This response was inhibited in the presence of neutralizing anti-CD4 monoclonal antibody GK1.5 (10 μg/ml; not shown) indicating that the main cell population reacting with the RNP1-U1A peptide 47–59 does correspond to CD4+ T cells.
Reactivity of rabbit antibodies to peptide 131–151 of the U1-70K protein with RNP1-peptide
Antibodies generated in outbred rabbits to peptide 131–151 of the U1-70K protein react with the cognate protein and with other spliceosomal proteins
We then questioned whether antibodies raised to peptide 131–151 are able to recognize the whole cognate protein and other proteins present in the same spliceosomal particle (Figure 3b). Ponceau red staining of membranes blotted with the three recombinant proteins U1-70K, U1-A, and U1-C and with the long synthetic fragment 21–119 of SmD1 revealed a single band at 65 kD, 35 kD, 20 kD and 12 kD, respectively, as determined with MW markers (Figure 3b, lanes 1–4). IgG antibodies from the rabbit immunized against peptide 131–151 in CFA (see above) strongly reacted with the U1-70K (lane 5) and the U1-A (lane 6) proteins, as well as with the U1-C (lane 7) and SmD1 (lane 8) proteins, which do not contain any RNP1 motif, suggesting the establishment of a heterologous B-cell epitope spreading phenomenon, and not solely immune cross-reactivity. No reactivity was found with SmBB' protein (not shown) or with any of the five proteins when tested with the serum from an outbred rabbit that received several injections of CFA alone (lanes 9–12). This rabbit did not develop any IgG reactivity to dsDNA (as measured by ELISA), and no clinical sign of autoimmunity was measurable 8 weeks after the last of three peptide administrations (result confirmed in several outbred rabbits). New Zealand White rabbits were not used in this set of experiments because they have a natural tendency to produce auto-antibodies [21]. A similar result was obtained with two outbred mice (ICR, Harlan) immunized with peptide 131–151, for which antisera react in Western immunoblot with the U1-70K protein and also with U1-A and U1-C proteins (data not shown).
Effect of P140 therapy on spontaneous T cell spreading
As U1-A and hnRNP-A2 peptides do contain an RNP1 motif, we could not rule out the possibility that the observed drop of responsiveness was simply due to the extinction of P140-specific T cells by a homologous-like, cross-reactive effect. To ensure that P140 treatment was leading to a true (non-cross-reactive) mechanism of 'intermolecular tolerance spreading', the goal was thus to demonstrate that T cell response to spliceosomal proteins that do not contain an RNP1 motif was also abolished in P140-treated MRL/lpr mice. U1-C and SmD1 proteins, which are associated to the U1-snRNP particle and are devoid of RNP1 motif, were appropriate candidates in this context. Epitopes recognized by CD4+ T cells are not known in U1-C protein. However, an epitope recognized by T cells from SLE patients and (NZBxNZW)F1 mice has been identified in the C-terminal end of SmD1 [22, 23]. This epitope is present in peptide 83–119, which encompasses the sequence 97–119 shown earlier in our laboratory to contain a B-cell epitope recognized by IgG antibodies from MRL/lpr and (NZBxNZW)F1 lupus mice and from patients with lupus [3, 10, 15]. In preliminary experiments with peptide dose-response measurements, we found that SmD1 peptide 97–119, but not SmD1 peptide 77–96, was efficiently recognized by MRL/lpr PBLs (not shown). With this new target, as above, we analyzed the reactivity of MRL/lpr PBLs (the peptide 77–96 was used as control) after P140 treatment. As shown in Figure 4, we observed that PBLs from P140-treated MRL/lpr mice did no longer proliferate ex vivo in response to SmD1 peptide 97–119 used as recall antigen in the culture (81% decrease; P = 0.0005).
A crucial question in our understanding of SLE is to identify the early cellular and molecular events that predispose to the loss of tolerance in individuals and those that trigger the transition from a preclinical status to overt disease and perpetuate the immune-mediated, inflammatory response. The concept of the T-B cell spreading phenomenon based on an initial acute autoimmune response restricted to a few epitopes using a limited number of T-cell clones for expansion is now widely accepted. However, the possible initiator antigens and the mechanism that render this(these) antigen(s) immunogenic in a predisposed context is not clear [1, 24–26]. It is also not clear what the link is that drives T-B cell diversification from these putative initial epitopes to a particular set of antigens. Our hypothesis is that the RNP1 motif, which is present in a number of RNA-binding proteins and represents a frequent target for B- and T-cell response, might be central in this process and partly explain autoimmunity to RNP proteins in lupus [12, 13].
In this model, the RNP1 motif initiates the spreading of the immune response to the whole protein (intramolecular spreading), then proceeds in an ordered manner to other proteins containing the RNP1 motif, and finally, to proteins that do not contain any RNP1 motif but are colocalized in the same particle (intermolecular spreading). In our previous work [14], we clearly demonstrated that the first step of this process effectively occurs in MRL/lpr lupus mice, in which the immune response diversifies from the RNP1 motif to the whole U1-70K protein (intramolecular spreading). In the present study, we validated the second step of our proposed model [13] by demonstrating that an immune response induced against the RNP1 motif can drive the diversification to other spliceosomal proteins that may or may not contain an RNP1 motif, such as U1A, hnRNP-A2, U1C and SmD1 proteins (intermolecular spreading).
Moreover, we not only confirmed that the sequence 131–151 of the U1-70K protein contains a recurrent epitope recognized by CD4+ T cells from MRL/lpr mice, but we also showed that the RNP1 motif present in other spliceosomal proteins can be recognized by MRL/lpr PBLs. Clearly, the RNP1 sequence 47RSLKMRGQAFVIF59 present within the U1-A protein induced a stronger proliferation of MRL/lpr PBLs compared to other RNP1 sequences, which were much less efficient and for some, very poor inducers. It has to be noted that the T cell epitopes of U1-A characterized previously by others [27–29] in MRL/lpr and (NZBxNZW)F1 mice and patients with SLE or mixed-connective tissue disease did not or only partially encompass the RNP1 motif. Epitopes recognized by autoreactive CD4+ T cells either in patients or in murine models of lupus or rheumatoid arthritis are not known in the case of the hnRNP-A2 protein.
The RNP1 motif is present in B-cell epitopes recognized in different proteins by antibodies from autoimmune mice and patients [[13]; and references therein]. In the present study, we show that antibodies elicited after immunizing a rabbit with the peptide 131–151 of the U1-70K protein cross-react with the short RNP1 motif contained within the immunizing sequence but significantly less well with the RNP1 motifs of U1-A and hnRNP-A2 proteins, indicating that in solution these short peptides do adopt different conformations. In Western immunoblotting, antibodies from this immunized outbred rabbit reacted strongly with the cognate protein U1-70K and with U1-A, but also with U1-C and SmD1, which are both devoid of RNP1 motif. These results fit well with the previous findings from several groups, which showed similar intermolecular antibody diversification in longitudinal studies of lupus patients and mice [1, 30, 31]. However, in our rabbits immunized with the 131–151 peptide, we found no antibodies to dsDNA or chromatin, no antibodies to recombinant SmBB', and no clinical sign of autoimmunity. Immunization of experimental rabbits and mouse models with the proline-rich peptide PPPGMRPP, another key sequence, which is present in several spliceosomal proteins and was reported to trigger spreading in immunized animals [32, 33], gave contradictory results in independent studies regarding the presence or not of anti-DNA antibodies and signs of lupus-like autoimmunity [34–36]. To our knowledge, PPPGMRPP peptide administrated in lupus mice was never shown to possess protective or tolerogenic properties.
The reasons that underlie the successful treatment of MRL/lpr mice with peptide P140, which significantly retards the emergence of dsDNA IgG antibodies and the progression of nephritis and prolongs the survival of treated mice, are not yet fully elucidated. We showed here that P140 administration not only abolishes T cell intramolecular spreading to other regions of the cognate U1-70K protein, but also leads to an impressive unresponsiveness of PBLs reacting with RNP1-peptides or longer peptides containing the motif, and with a peptide from the C terminus of SmD1 protein, newly characterized in this study and originated from a spliceosomal protein that does not contain any RNP1 motif. Thus, in treated animals, P140 peptide elicits clearly a state of bystander suppression leading to the downregulation of autoreactive T- and B-cell response to other self-antigens (so-called tolerance spreading [37]). Important questions remain at this stage as to whether P140 peptide induces initial tolerance by playing the role of antagonist or partial agonist of the receptor of autoreactive T cells, or by using another mechanism, as previously discussed [9]. The fact that P140-treated animals could mount a normal immune response to foreign antigens is not an unexpected result if we consider that P140 peptide primarily target the TCR of specific autoreactive T cells.
= enzyme-linked immunosorbent assay
= Freund's adjuvant
= peripheral blood lymphocyte
= systemic lupus erythematosus
This work was supported by CNRS and a grant from the Fondation pour la Recherche Médicale. VP was supported by a fellowship from the Association de Recherche sur la Polyarthrite (ARP). We thank M. Valette (Faculté de Médecine RTH Laennec, Lyon, France) for providing influenza virus (strain A/NT/60/68).
Authors' Affiliations
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Those look so good. Nevermind the 4.0/5.0s...I want the 3s.
The 3.0s look damn good. Might even get that CW.


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So, do I need tanks???
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Topic: So, do I need tanks??? (Read 4905 times)
Hero Member
Posts: 3508
MCI 1995 DL3, DD S60, Allison B500.
So, do I need tanks???
May 14, 2009, 08:17:12 AM »
Actually, I think that we have made up our minds. We have decided to put tanks into our bus
. I have read the archives, on the subject, but still have some questions.
Here is what we need. I am looking at what sizes to get. Often when we travel, there will be ten people on board (large family). We have considered at least a 200 gallon fresh water, and don't know how to split the gray and the black water.
My questions...Is 200 fresh enough? Where do we put it? We were thinking about putting it under the bed in the back, but thought that much weight behind the axles might be a little much (not to mention we will be pulling a cargo trailer that is 16'/8' with a 14K GVWR). We would like to get all of our tanks in the third bay, but it will be a VERY tight squeeze. We are getting rid of the OTR AC so that will give us a little extra wiggle room. The third bay is 8 feet long, so it is our biggest bay. We also have a 20 KW generator in their (which is good sized), and 2 trace inverters (stacked). We will move our batteries into the place where the AC coils were, and save the outer part where condenser was, for a backup 8KW generator. Do we need one or two water pumps (I am thinking two, just because it would be a backup).
Also, what kind of toilet should we get. I thought that I heard Microphor (maybe?) was recommended, but I don't remember. We want to get high quality stuff. And where do we get a shower? Do you buy them, or make you own?
Any advice, or criticism, on these issues, would be GREATLY appreciated. Some of you guys have a wealth of information, and I would love some of your ideas. This is our first, and it is a little tough.
Thanks in advance,
God bless,
Last Edit: May 14, 2009, 08:20:41 AM by John316
Sold - MCI 1995 DL3. DD S60 with a Allison B500.
Re: So, do I need tanks???
Reply #1 on:
May 14, 2009, 08:33:27 AM »
As you already stated, ALL of the answers to your questions are in the archives and a little leg work on your part will put the answers in front of you! These things have been discussed over and over so if they keep being brought up, why keep archives?
My opinion!
Hero Member
Posts: 3508
MCI 1995 DL3, DD S60, Allison B500.
Re: So, do I need tanks???
Reply #2 on:
May 14, 2009, 08:43:06 AM »
Thanks for your post. I have been looking through the archives. I think that search feature is what is a little lacking. But I will keep trying. From what I have seen so far, my questions here still weren't answered.
God bless,
Sold - MCI 1995 DL3. DD S60 with a Allison B500.
Hero Member
Posts: 8073
Re: So, do I need tanks???
Reply #3 on:
May 14, 2009, 08:51:49 AM »
With my wife and I, we can go a week on 130 gal of fresh. That works out to be 9.3 gal per day per person (this is taking Navy showers and washing dishes). I think if you used 8 gal a day per person, since little people take less to shower, that would be a good rule. So 200 gal would get almost 3 days of dry camping for you.
When I was truck driving I had a 20 gal toilet tank that lasted one week. That translates into 2.8 gal per day per person, or 28 gal per day for you. So to keep with the same 3 day schedule, that would be a 84 gal black tank, and with simple subtraction from 200, a 116 gal gray water tank to also get you through the 3 days (would use more like 130 gal).
For my shower I used a Kohler 36"x36" shower pan that has the drain in one corner since my shower is mounted right where the floor raises up a bit. Then built the walls of 3/4" plywood with fiberglass panels over the plywood.
Toilet wise, I have a Sealand 510 toilet. Looks good, but the bowl isn't steep enough-sometimes have to help the solids to go down. I would stay with a simple gravity RV type toilet. Macerators are nice, but I see alot of maintenance with 10 on board. Good Luck, TomC
Tom & Donna Christman. '77 AMGeneral 10240B; 8V-71TATAIC V730.
Sr. Member
Posts: 494
Re: So, do I need tanks???
Reply #4 on:
May 14, 2009, 08:53:01 AM »
I think a lot of this is personal preferance. How often do you want to stop to dump, fillup water etc. Have to weighed your bus? Do you know how much room (weight) you have for these options. As far as the shower goes once again personal preferance, but I do know to stay away from any of the house hold showers with that sheeted backing you glue on the wall. It does crack in extreme weather conditions or temp. changes
Have Fun!!
Hero Member
Posts: 779
Re: So, do I need tanks???
Reply #5 on:
May 14, 2009, 08:54:22 AM »
To provide a reasonable estimate for your tank size, we not only need to know the number of people, but how you plan to use the bus. Basically need to know how long from fill-up & dump, until the next fill-up & dump. 3 days? 3 weeks? You may have already covered this in your previous threads, and I'm sorry that I don't recall.
But in general terms, a 200 gallon fresh water tank is a pretty healthy size. Mine is close to that and even if I am boondocking for a couple of days (family of 5) I never fill it more than half full. If everyone wanted a daily shower, that might be different.
Conventional logic usually sizes the black and gray tanks to total up slightly more than the fresh water capacity. The percentage split between the two depends on how the bus will be used. Personally, I tend to favor making the black water tank slightly larger. Two reasons for this: First, my gray water tank is plumbed to drain through the black water tank before exiting out the drain hose. If I found myself running out of gray water tank and still had room in the black, I can open the valve for a few seconds and bleed gray into black (but not vice versa). Second, gray water is easier to get rid of when remotely boondocking. When washing dishes, you can set up two dish pans, one to wash and one for final rinse. The sudsy wash water with it's food scraps goes into your gray tank, while the dishpan with still clean rinse water gets flung in the bushes or emptied down a storm drain.
I don't at all condone emptying your gray tank on the ground. But depending on where you are, I wouldn't have a problem with taking the water left over from cooking pasta (no food pieces) and finding a way to return it to nature. All this adds up to saving gray tank space.
If you're going to be dumb, you gotta be tough.
Hero Member
Posts: 3508
MCI 1995 DL3, DD S60, Allison B500.
Re: So, do I need tanks???
Reply #6 on:
May 14, 2009, 09:15:00 AM »
Thank you all so much. I have been continued to search the archives, but these answers are great!
TomC, thanks for the info. It really helps a ton. We are planning on having a regular RV toilet, and we are also considering putting a second toilet in the shower room (we are going to have the bathroom split up). The second toilet in the shower room would have a pump on it, and would only be used for emergencies (we have had the stomach flu all at once, and that includes the runs
, at that point I thought a backup toilet with a pump would be nice for emergencies).
Grant, no we haven't weighed the bus. Good idea though, we will do that when we pick the bus up (we are getting Jakes on it finally). Thanks for the tip about staying away from the shower wall material. We won't go with that, as we do go up to extreme weather climates (but not all the time, just occasionally).
Wec, we would like to go as long as possible. I would think that a week would be fine under "normal" circumstances, but longer would be nice. It all depends on what we are doing on a certain trip. If we are around "polite" company, then we take daily showers (at least the girls do
). We don't have any tanks right now, so that is why I am trying to figure out what to do. I agree with you, if it is just a camping vacation, I think that the 200 would hold us just fine. But, I am thinking that we need more....
Thank you all so much!!! Keep it coming, and I appreciate it greatly.
God bless,
Sold - MCI 1995 DL3. DD S60 with a Allison B500.
Hero Member
Posts: 551
'60 PD4104-4355(sold) Now Freightliner Conversion
Re: So, do I need tanks???
Reply #7 on:
May 14, 2009, 09:29:27 AM »
We get by with 5 gallons/day per person. "Navy shower" every day. Water for drinking and coffee is bottled water ($0.15-0.25/gallon at water machines). To control shower water we use a "sink sprayer" unit. Simple and effective. Have been doing this for 24 years. We are 1/2 timers for the past 10 years. Works for us.
Gary D
USBUSIN was our 1960 PD4104 for 16 years Ustruckin' is our 2001 Freightliner truck conversion
Hero Member
Posts: 3512
NEWELL in South Carolina
Re: So, do I need tanks???
Reply #8 on:
May 14, 2009, 09:54:27 AM »
Since you want to go as long as possible, fill all the available space with tanks. Then your actual usage will determine the how long that is.
Extra tank capacity is easier to deal with than extra black water.
A gallon of water weighs ~8.4 lbs, so 200 gallons ~ 1680 lbs.
I'd suggest you check out sheet metal fabricators in your area & get estimates. The fresh water tank should be "food grade", the black & grey won't matter.
Plastic tanks are usually wider than tall & of such a size that baffling isn't usually required.
My preference is to build custom tanks out of 14GA stainless steel. The tanks will be vertical & baffled. I will use ball valves for the drains. The vertical tanks offer space advantages & the added fluid height aids in faster tank dumping.
If your toilet is directly over the black tank, you will greatly reduce the amount of water required to flush. That is what we have in the TT, & the 4 of us have gone 5 days & didn't fill a 30 gal black tank, the 30 gal grey required dumping after 3 days (which included 7 showers). needless to say, we are very conservative with water.
Life is all about finding people who are your kind of crazy
Get your facts first, then you can distort them as you please (Mark Twain)
Education costs money. But then so does ignorance. (Sir Claus Moser)
Hero Member
Posts: 3508
MCI 1995 DL3, DD S60, Allison B500.
Re: So, do I need tanks???
Reply #9 on:
May 14, 2009, 10:08:43 AM »
Gary, thanks for the insightful post.
Kyle, interesting. I kinda like that idea, since that maximizes the space. With stainless steel, what do you use for tank monitors? Our main toilet will be directly over the black tank.
God bless,
Sold - MCI 1995 DL3. DD S60 with a Allison B500.
Hero Member
Posts: 1036
Re: So, do I need tanks???
Reply #10 on:
May 14, 2009, 10:16:40 AM »
John, I think you are going WAY overboard for "backups", you will be dragging that "stuff" up and down hills, maintinence, lack of room elsewhere, ect, ect. With a family of ten you ARE going to need space for other things that are more neccessary
than backup. Your present thinking IMHO is going to run you out of space.>>>Dan
EX 4106 (presently SOB)
Cedar City, Ut.
72 VW Baja towed
Confidently Ignorant
Sr. Member
Posts: 407
Re: So, do I need tanks???
Reply #11 on:
May 14, 2009, 10:57:43 AM »
Quote from: John316 on May 14, 2009, 08:43:06 AM
Thanks for your post. I have been looking through the archives. I think that search feature is what is a little lacking. But I will keep trying. From what I have seen so far, my questions here still weren't answered.
God bless,
As a new member I feel your pain. I've searched for many different subjects and the search returns 30 pages of posts but most aren't relevant or they don't answer it from my needs. I would find it helpful if an experienced member would post a link to a thread they've found successful. A simple example, I was looking for jacks and many results related to Jack Conrad. He's a nice guy but I didn't contact him as I'm pretty sure he's not going to ride around with me ready to lift my bus when needed.
I didn't notice if you were planning on camping with hookups. If you mix hookup sites into your camping trips, 200 gallons is plenty not to mention heavy. If you are worried about your family taking too long in the shower keep your hot water tank very small. It will only take a short time before everybody understands the need for speed.
Sold 85 Neoplan 33ft 6V92ta, sadly busless
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Re: So, do I need tanks???
Reply #12 on:
May 14, 2009, 11:03:15 AM »
John, we have 180 gal fresh water tank 120 gal gray and 60 gals black works good for us.The Micophor is a good investment at 1000 + bucks just be sure and buy the electric flush not the air.I used a stainless tank one time for fresh water and could never get the metal taste out of the water. good luck
Life is short drink the good wine first
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MCI 1995 DL3, DD S60, Allison B500.
Re: So, do I need tanks???
Reply #13 on:
May 14, 2009, 11:04:50 AM »
I wonder though, are we? If a water pump goes out, then everybody is out of water. Now that is somewhat okay for a couple, but a large family, that has to be somewhere that night, is a completely separate story.
On the extra toilet, that was something that I thought would be nice, if possible. I don't have to go into details to tell you guys how rough it can get with everybody having the stomach flu
Thanks for the post, Dan. You have some good food for thought. I think that we are going to plan on pulling a trailer anyways (except for a little week vacation or something).
Mike, I agree, Jack Conrad is a nice guy, just maybe not strong enough
. We don't plan on using campgrounds, basically at all. We will dump, and fill up at Flying J's (or similar), and we will stay at free parking places, church parking lots, and at our host's houses.
Clifford, thank you for the post. Interesting about the SS tank....
God bless,
God bless,
Sold - MCI 1995 DL3. DD S60 with a Allison B500.
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Posts: 808
Re: So, do I need tanks???
Reply #14 on:
May 14, 2009, 11:23:05 AM »
I would make mockups out of plywood and make or purchase tanks that fit what space I have available. I'd let that determine how many gallons of each, fresh, grey and black would be as well as how to run the various fill/drain plumbing. My black sits on top of my grey and that really helps the gravity flow for discharge.
I had mine done professionally and it completely filled the rear cargo bay on the Eagle. They used 3 80's for fresh. (No slosh or movement with smaller tanks hooked togather). The black is 68 and when close to full puts enough pressure on the dump valve to make a new one drip when it is close to full, so if I want more I would need a better valve than the common ones used.
My grey is stainless 160 gallon and seems sufficient. I also have alot of people on often and I have showered slept and fed up to 17 people a day. Everyone showers everyday on my coach, or they walk! I could go 3 full days but needed service on the 4th day to give you an idea. I use 1 pump and just carry a new spare, they are very easy to repair, just make sure they are very accessable so you don't have to stand on your head to fix one.
I'd move the genset, but there are other ways. Just my preference. Low and heavy in that bay is good in bad weather, but it really doesn't matter what is low and heavy, I just chose the water. I would try to keep it off the steer axle though if I could.
200 gallons in the living space could go through the windshield in an accident, I have seen people do it, but I don't know how to really secure it and I personally run alot of snow and ice.
A friend and I were just out playing in California for 15 days. I was parked on a farm so I let the grey run on the pasture. The black was about 1/2 full, and I refilled the fresh once and brought most of the refill home.
I will probably do Burning Man for a week this fall and will take a couple of 55 gallon drums extra of fresh to pump into my tanks to help the ladies in the area out. (Burning Man is all about trading). As big as your trailer is maybe some water would be OK back there when you need extra.
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$1T in $1000 bills = 142 miles high
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Ski Merino Socks
Skiers of all levels
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Cores disponíveis:
Now you can finally get ski socks that don't make you choose between sweating (because they are heat insulating) and freezing (because they are breathable). Your feet stay dry and your legs enjoy a pleasant climate. No matter what the weather conditions: -15 °C (5 °F) on the glacier or +5 °C (+40 °F) in the valley. This is made possible by the clever combination of merino wool and water-repellent synthetic fibers for perfect heat and moisture management. Add the protective padding and deliberately thinner compression knitting and you've got an innovative product that delivers superior comfort and impressive performance.Mais detalhes
Ski Merino Socks
Atributos de qualidade
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Atributos funcionais
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67 % polyamide, 23 % wool (merino),
10 % spandex
Made in Germany


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Home > Company Profile
Guangzhou Newsky Electronic Co., Ltd.
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The supplier's company premises has been checked by Alibaba.com staff to ensure onsite operations exist there. A third-party verification company has confirmed the legal status of the supplier.
|Business Type:
|Trading Company
|Guangdong, China (Mainland)
|Main Products:
|video game parts and accessories,cellphone parts and accessories
|Total Employees:
|11 - 50 People
|Total Annual Revenue:
|US$1 Million - US$2.5 Million
|Year Established:
|Top 3 Markets:
|Western Europe 30.00% North America 30.00% South America 10.00%
Guangzhou Newsky Electronic Co., Ltd. was established in 2000, engaged in the foreign trade of spare...
Guangzhou Newsky Electronic Co., Ltd. was established in 2000, engaged in the foreign trade of spare parts of smart phones, game consoles of various international brands. We successfully agent built-in accessories and peripheral devices for APPLE, HTC, SAMSUNG, BLACKBERRY, SONY, NINTENDO,SEGA, XBOX, etc.
Our office and shop are located in Guangzhou, with warehouse in Hong Kong. We are a harmonious and vibrant team with members at the average age of 28.88. We have a perfect company system and adopt human-based management. With lofty goals, we have been in steady progress.
Guangzhou Newsky Electronic Co., Ltd. enjoys support and recognition from customers. We supply original, assembled and second-hand accessories for global electronic digital parts shops, maintenance centers and repair markets. Customers can make multiple choices from our products. We have excellent reputations in this business, and clients have strong viscosity with us.
Now we carry forward our cause and forge ahead into the future, changing from product-oriented to service-oriented. We serve our worldwide customers with integrity.
Products and services:
LCD screen, touch screen, LCD assembly, flex, button, laser lens, lens, drive, mainboard, battery, AC, digital cable, OBD2, etc.
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Trade & Market
|Main Markets
|Total Revenue(%)
|Western Europe
|North America
|South America
|Eastern Europe
|Mid East
|Southern Europe
|South Asia
|Northern Europe
|Central America
|Southeast Asia
|Export Percentage:
|91% - 100%
|Export Mode:
|Using an agent
|No. of Employees in Trade Department:
|11-20 People
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Hotels in Nykoping
Hotel reservation
Hotel rooms in Nykoping
There are not so many hotel rooms in Nykoping, just 3, still everyone can find an offer to suit his choice. The most common offers in this city are hotels, they count 1 out of 3, and their prevalent location is central district. According to our practice, it is the most profitable to book a hotel in the north of Nykoping, as it is the location of the hotels with optimal cost and quality offers along with the cheap hotels. Taking into account the small size of the city, there is no need to dwell hotel reservation on the location: the road to the city center would take less then half an hour, so one can book a hotel in any district of Nykoping. The most populated with hotels district of Nykoping is the south, 33% of cheap hotel rooms are concentrated here. However, even those who want to live in the center can choose from 1 variants of hotel rooms. Nykoping is an ideal place for family and couple, such travellers rate hotels there 8.4 out of 10 on average. Apart from all these, hotels in Nykoping offer their customers laundry service, pets allowed, spa and business centre. The most appreciated amenities in Nykoping hotels are wellness and wifi. According to the visitors reviews, hotels in Nykoping has great amenities, great room and clean. Take a look at our selection of the cheap hotel rooms in Nykoping to make a well-informed choice for your reservation!
Best hotels in Nykoping
Hotels in Nykoping are spread around the city within the distance of 6.3 kilometers, while 1 hotels are located directly in the city center. Those who are keen on antique should definitely fall in love with
Blommenhof Hotel
, the oldest hotel of Nykoping, constructed in 1908. This hotel is located 1.4 km away from the city center. The most popular accommodations in Nykoping is
Blommenhof Hotel
, 4 stars hotel located in a 1.4 kilometers from the city center.
The best hotels are here and now. Try them!
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Is Waikato Slate & Tile Centre incorrect or missing key business information? Update your Hotfrog listing today.
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Hotfrog NewZealand provides information regarding Waikato Slate & Tile Centre in Cambridge Waikato. Waikato Slate & Tile Centre is located at 24 Pope Terrace, Cambridge Waikato 3432 and provides Home Improvement services. Contact them on 07-827 8925 or by visiting them on their website http://www.waikatotiles.8m.com.
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2001 BMW 540i Repair and Maintenance Costs
A guide to repairs, service, and maintenance costs for your 2001 BMW 540i
Never overpay. Check prices with our estimator
Verified by mechanics, reported by drivers like you
Ask a mechanic in our community
2001 BMW 540i ProblemsEngine Oil Leaks
Engine oil leaks commonly develop from both valve cover gaskets, intake manifold, and PCV plate after about 60,000 miles.
Leaks are known to develop in the cooling system. It is common to see more than one part recommended for repair/replacement due to overlapping labor and the use of many plastic parts in the system.
The blower final stage fan resistor is known to fail. Symptoms are the heater fan speed varies regardless of the setting, the fan will not turn off when the car is switched off, or the fan will not switch on at all.
2001 BMW 540i QuestionsMy car engines was leaking oil but now it's shaking and losing power (1 answer)
is this a indication of engine problems?
Everything is working but there is no air coming out at all
The steering on my 2001 540iA is acting up. It is somwhat harder to turn to the right, gradually getting easier as the car warms up. There is also a suspicious "whirring" sound--sort of like it wants more fluid. I'd like to do the KISS process and first check the fluid to see if i...


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Explore Eagles Alaska, Alaska Usa and more!
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Flying Eagle - Colorful sky~Animal Spirit Guide: The Eagle - "In Nat. Am. Cultures the Eagle is Sacred as a Spiritual Messenger one that delivers peace, fertility, honor, freedom, strength, illumination, courage & the ability to rise above all. Bald Eagle Spirit Medicine is to teach spirit to see the HIGHEST TRUTH... In journey work, Bald Eagle may escort you to distant realms... Expert at Soul Retrieval.


text: Gay marriage. Right or wrong? According to Chick-fil-A's Dan Cathy and many Americans of faith, the Bible has the final word. Marriage = man and woman. End of story. This simple statement, and still widely held belief, has infuriated homosexual activists and become a focal point of the culture war that is engulfing America. Gay marriage zealots, who claim to ...Read More »


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The Latest Lemon Grove, CA News
Lemon Grove is a city in San Diego County, California. The community was settled in 1869 by its first known permanent resident, sheep rancher Robert Allison. From the 1860s to 1900 the area was known for sheep and poultry ranching and vegetable crops. In the early 1900s, the community acquired its name when large citrus groves were planted, and lemon and orange growing and shipping became the area's major industry. There is a ten-foot-wide statue of a lemon located at 3361 Main Street. It lies before a small lemon grove beside the Orange Line Trolley tracks near the local Trolley station and downtown bus stop. Written across the base of this monument are the words "Best Climate On Earth".
Lemon Grove, CA Real Estate
Looking to Sell Your Lemon Grove, CA Home? Our local Lemon Grove, CA Real Estate Agents can provide you with a free market report on your home along with a customized listing proposal to get your home sold fast!
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If you're searching for a home in San Diego (our you just want to check out what's happening), our real estate listing is like no other. Our customized "Features Search" allows you the ability to browse listings based off of things like "Open Floor Plan, Natural Light, Large Backyard, etc." You can begin browsing Lemon Grove, CA Real Estate Listings including Homes For Sale and Commercial Real Estate in the Lemon Grove, CA Neighborhood of San Diego for free in our Lemon Grove, CA Real Estate section.
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As a property owner, we know you want to keep track of how your property is doing in the marketplace. Want to know how much your Lemon Grove, CA Home is Worth today? Our automated home valuation tool just needs property address in order to provide you with Instant Home Valuation. In addition to your automated value, one of our local Lemon Grove, CA REALTORS will provide you with an in-depth market report which includes expected rental income, expenses, cap rates and investment potential of any Lemon Grove, CA property.
Obtain a free complimentary automated home value report for your property in Lemon Grove, CA. This report draws on public records data, Zillow data, as well as recent Real Estate sales in Lemon Grove, CA and will be displayed instantly. Find Your Lemon Grove, CA Home Value Now!
Lemon Grove, CA Mortgage and Home Loans
Working with the right mortgage lender is a crucial part of purchasing a property. Sometimes when you use a large bank, you don't get the attention you deserve which can lead to issues during the closing process which can cost you thousands. The San Diego Connection network together with The Local Realty have partnered with local Lemon Grove, CA lenders to offer our subscribers the lowest mortgage rates in the industry along with the best possible customer service. If you're considering purchasing or refinancing a property in Lemon Grove, CA, call (866) 538-6057 to be matched with some of our preferred local mortgage lenders.
Selling Real Estate in Lemon Grove, CA
The San Diego Connection Networks online Real Estate presence offers vast exposure buyers of San Diego Real Estate. Our email list alone includes over 20,000 local residence and our online marketing reaches millions of potential buyers across the world. Each month we generated over 300 new registered users which search homes in all San Diego communities. Our team of Experts at The Local Realty understands real estate marketing and how to sell your San Diego home for top dollar. If you're considering Selling your home in San Diego, Contact the team at The Local Realty to find out how they can help sell your home and maximize your equity.
Lemon Grove, CA Short Sales
If you are having trouble paying your mortgage a short sale may be the best option for you. Many San Diego home owners don't want to short sale for fear that they 1) won't be able to buy another home OR 2) It will hurt their credit. However, through a strategic short sale an expert San Diego short sale agent can help protect your credit and even get you into a new home! Our San Diego Short Sale Agents have helped over 300 homeowners avoid foreclosure since 2007. Talk with one of our experts at The Local Realty in San Diego for a free consultation by calling (866) 538-6057
Lemon Grove, CA Relocation Services
We offer professional relocation services for companies and individuals relocating into or out of San Diego County.
VA Approved Condos in Lemon Grove, CA
With San Diego being one of the largest military towns in the country, you may be looking to use your VA benefits by purchasing a home in San Diego using a VA loan. Did you know that not all properties are approved for VA loans? Don't worry, we've got you covered. Through our partnership with VA Home Map, we offer a refined list of VA approved condos and homes throughout San Diego. We also know what you need as an active duty military member in San Diego! We have helped hundreds of military members purchase a home in San Diego. Whether you need to close on a home while you're deployed, or you're in training for a week with no access to phones, we have the procedures in place to make sure you can close on a home without the headache. Speak with a VA loan expert and training Military Relocation professional with The Local Realty in San Diego to learn more.
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Bologna thought they had fought back to beat Torino, but Rolando Bianchi snatched a 2-2 draw with the last kick of the game.
Click here for the full match report.
Keep up to date with the latest news and action from Spain's Primera Division with Football Espana - from the team behind Football Italia.


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Introducing Person 2 Person payment services!
Bank of Blue Valley
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Pippo Inzaghi is furious at being left off Milan's Champions League list and could return to his former club Atalanta in January.
The striker has been out for several months with a knee injury and, although he has now recovered, Coach Massimiliano Allegri opted to leave him out of the UEFA list.
Allegri has confirmed Inzaghi was angry at the decision and it is now reported they could part ways in January.
Atalanta would be willing to bring SuperPippo back into the fold, where he scored 24 goals in 33 Serie A games in 1996-97.
Another of the 38-year-old's former clubs, Parma, are also interested in giving him another chance.
Keep up to date with the latest news and action from Spain's Primera Division with Football Espana - from the team behind Football Italia. [11]


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This regional capacity development technical assistance (TA) project seeks to maintain the Asian Development Bank's (ADB) efficiency and effectiveness to support better project implementation in accordance with ADB's midterm review of Strategy 2020. This TA is providing consultants to provide training and on-the-job assistance during key stages of procurement and project implementation.
Latest Project Documents
Consulting Notices See also: CMS
|International Procurement and Capacity Development Specialist (AFG/TKM)
|2 Mar 2017
Procurement Notices See also: Operational Procurement
No notices are currently available for this project.
Procurement Documents
None currently available.
Project Data Sheets (PDS) contain summary information on the project or program. Because the PDS is a work in progress, some information may not be included in its initial version but will be added as it becomes available. Information about proposed projects is tentative and indicative.
The Public Communications Policy (PCP) recognizes that transparency and accountability are essential to development effectiveness. It establishes the disclosure requirements for documents and information ADB produces or requires to be produced.
The Accountability Mechanism provides a forum where people adversely affected by ADB-assisted projects can voice and seek solutions to their problems and report alleged noncompliance of ADB's operational policies and procedures.
In preparing any country program or strategy, financing any project, or by making any designation of, or reference to, a particular territory or geographic area in this document, the Asian Development Bank does not intend to make any judgments as to the legal or other status of any territory or area.


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B2B CMS Responsive Website
This project received 38 bids from talented freelancers with an average bid price of $606 USD.Get free quotes for a project like this
Project Budget$250 - $750 USD
Total Bids38
Project Description
We are a direct sales company with full time independent sales agents which is mainly b2C. We are also starting to grow our B2B business with telesales in our call center offering a range of products,services & solutions to the small to medium size offline business ( websites,mobile sites, text messaging....) We also plan to host all websites we sell to our clients . We currently have run some 3rd party cloud software that would ideally be integrated into the new cms responsive website. We will need to cater for clients login portals. To cater for the B2C business a social page . photo galleries. The content management system would be the central hub for all content distribution reformatting & content for the different platforms ...mobile , desktop .
I'm looking for a modern clean design with priority for possible integration with 3rd party platforms. I will also require the the services of a very good copy writer for B2B type scripts. Our current site [url removed, login to view]
This will be a new website from start to finish.
Hourly - Est. Time Less than 1 month, Full-time - 30+ hrs/week
Skills Required
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- Set your budget and the timeframe
- Outline your proposal
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The NSO performs the Russian master's Sérénade mélancolique and Valse-Scherzo--both featuring NSO Concertmaster Nurit Bar-Josef--plus Tchaikovsky's "Winter Dreams" Symphony and Hamlet Fantasy Overture.
Please use the event calendar to search for current events.
All events and artists subject to change without prior notice.
All ticket prices are subject to change based on demand. Purchase early to lock in prices and the best seats!


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New Market Real Estate
New Market is located in Virginia. New Market, Virginia 22844 has a population of 2,312. New Market 22844 is less family-centric than the surrounding county with 20.31% of the households containing married families with children. The county average for households married with children is 26.65%.
The median household income in New Market, Virginia 22844 is $32,031. The median household income for the surrounding county is $49,953 compared to the national median of $53,046. The median age of people living in New Market 22844 is 41.7 years.
New Market Weather
The average high temperature in July is 87 degrees, with an average low temperature in January of 21.8 degrees. The average rainfall is approximately 35.5 inches per year, with 22.9 inches of snow per year. No weather information available.
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Give Now
Wilson coached the Hoosiers for six seasons. He will be co-offensive coordinator at Ohio State.
Athletic Director Fred Glass announced a plan Sunday to fund several athletic facilities as part of the University's bicentennial campaign.
Indiana University's athletic director says he expects more than 40,000 people to turn out for Thursday's game.
Fred Glass sent a new proposal to Kentucky University as to where the IU-UK games should be played.
The IU-Kentucky basketball rivalry may take a hiatus for several years.
IU athletics director Fred Glass says he wants to hire coaches on long-term contracts because it brings stability to their programs.
A recent study published by The Wall Street Journal ranked Indiana Men's basketball the third most valuable program
Indiana University will officially introduce Curt Miller as the new women's coach this afternoon.
IU Athletics and the Indiana State Police department are advising fans to take alternate routes so they don't miss kick off.
The approval to build the fields fulfills a promise Indiana University made to its baseball and softball programs several years ago.


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Newegg.com - A great place to buy computers, computer parts, electronics, software, accessories, and DVDs online. With great prices, fast shipping, and top-rated customer service - once you know, you Newegg.
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This review is from: Case Logic CDW-208 BLACK 208 Capacity CD Wallet
Pros: Newegg sells it.
Cons: Flimsy. At least I think flimsy. I have numerous different brand large (100+ and 200+) CD/DVD binders. I have not bought a new one in about 5 years. It just feels cheap. The special CaseLogic page system just seems weak when filled with DVD's. Hard to describe.
Other Thoughts: Shopping around, I have notice that the quality from most brands has diminished as well. Instead of tough nylon, we have settled for plastic and pleather which just seems flimsy and weak. Long term storage would be fine, but daily use, I not too sure.READ FULL REVIEW
This review is from: TOSHIBA Laptop Satellite Pro S300M-S2142 Intel Core 2 Duo P8600 (2.40 GHz) 2 GB Memory 160 GB HDD Intel GMA 4500MHD 14.1" Windows Vista Business with XP Pro downgrade media
Pros: I bought this to replace my old Dell C610. If you need a serial port and lots of other ports, go with this. Small enough for travel, decent battery life, placement of controls in good places. I don't really have a need for the fingerprint reader. I am pretty impressed with it's DVD burning capabilities.
Cons: Bloatware, but wasn't hard to cleanup. 2GB of RAM, but another easy fix. I wouldn't exactly call it lightweight, but not a brick either.
Other Thoughts: For the price, this IS a great deal. This is my first attempt at Vista Business, so far so good. As soon as I run into problems, I will downgrade. I have not had a chance to try any medium level resource games on it yet. NEWEGG!!!READ FULL REVIEW
This review is from: NEC Black 16X DVD+R 8X DVD+RW 8X DVD+R DL 16X DVD-R 6X DVD-RW 16X DVD-ROM 48X CD-R 32X CD-RW 48X CD-ROM 2M Cache IDE/ATAPI DVD Burner
Pros: I have four from the ND-35XX series, including this model, fantastic drive! I have burnt, ripped, and read all types of media. Have had this model for almost three years, and a ND-2500 for over five years, no problems.
Cons: Little spool-up noise, but not that I really care
Other Thoughts: It will be a sad day when these are not available. I recommend them to all my friends and family, from Newegg, of courseREAD FULL REVIEW


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Sonoco™ Insulated Shipper Multipurpose Containers and Accessories
Ideal shippers for the safe storage and transport of most refrigerated or frozen materials
Manufacturer: sonoco™ TS316
Description & Specifications
|Product Type
|Foam Insert
|Expanded Polystyrene (EPS) Foam
|Height (English) Exterior
|7.25 in.
|Height (Metric) Exterior
|Length (English) Exterior
|11 in.
|Length (Metric) Exterior
|Width (English) Exterior
|9 in.
|Width (Metric) Exterior
|For Use With
|Use for culture packs, serums, vaccines, and laboratory specimens; For dry ice sublimation.
|Dimensions (L x W x H) Exterior
|28 x 23 x 18cm


text:
Traditional Fit Non-Iron Dress Shirt (Online Only)
Product not available
Have questions?
Chat with us or call 1.888.282.6060.
HideDetails & Care
Graph checks add classic style to preppy spread-collar dress shirt crafted from non-iron cotton.
- Removable collar stays.
- French placket.
- Rounded, single-button cuffs.
- Chest pocket.
- Split back yoke.
- Cotton; machine wash.
- By Nordstrom; imported.
- Men's Furnishings.


text: 2006 Boise Fitness Expo Pictures - FAME Fitness Model Search. Page 1. By Bodybuilding Contests Last updated: Apr 10, 2006 The 2006 Bodybuilding.com Fitness Expo was huge especially with the addition of NPC and FAME contests. Page 1 Of 4 :: 53 Total Pictures CLICK TO ENLARGE ALL PICTURES! Gallery Navigation Main Page - Next Page » About The Author Bodybuilding Contests Check out our coverage of the latest bodybuilding, fitness, and figure contests! We have contest previews, schedules, results, and huge galleries... View All Articles By This Author Rate This Article


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Packed with information about machine modifications, metalworking equipment and gear reviews, it's the go-to guide for anyone who wants to learn more about this fascinating activity. Each issue is filled with advice on milling, drilling, finishing and sharpening tools. From micro machining to metal work on a lathe, The Home Shop Machinist teaches readers how to create successful projects. Whether you're looking for information about machine modifications, accessories for your metalworking equipment, shop improvement articles or step-by-step instructions, you'll find relevant advice in each new issue of The Home Shop Machinist.
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Nov 09, 2016 : Ford Motor Company has announced that it will develop a green and modern global technology and business center in Chennai, which will cater as a hub for mobility solutions, product development and busine ss services for India and other markets around the globe. The company plans to invest Rs 1,300 crore to further strengthen its connection with India and take the workforce count up to 12,000 employees in total. That will make the Chennai facility Ford's second-largest center of employment globally, after Dearborn, Michigan, US Read More Read Less
We get our hands on the massive new Ford Endeavour to see if the Americans can still get the big brash SUV formula right. Read on to find out....
Ford has launched the second generation Figo hatchback, which promises to be packed with equipment, safety features and looks a lot better than before. However, the compact car market has undergone a complete change in the recent past. Can the all-new Ford Figo carve a niche and recreate history...
We finally get our hands on the long awaited Ford Figo Aspire compact sedan. So does it have what it takes to take on the likes the Maruti Suzuki Swift Dzire, Honda Amaze and the Hyundai Xcent? Read on to find out.
The muscle car scored low marks in child occupant protection and safety assists
After reducing the prices just last September, Ford has hiked the prices of its flagship SUV after just three months
Pays homage to the legendary 1967 Shelby Super Snake
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