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Integrated Analysis of Innate, Th1, Th2, Th17, and Regulatory Cytokines Identifies Changes in Immune Polarisation Following Treatment of Human Schistosomiasis. | |
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PMID: 23045617 Owner: NLM Status: Publisher |
Background. Schistosomiasis elicits cross-regulatory immune responses, but it is unclear how antihelminthic treatment affects this balance. This study integrates data on 13 cytokines elicited by 3 schistosome to examine how praziquantel treatment alters immune polarization and whether post-treatment cytokine profiles influence reinfection status.Methods. Venous blood from 72 Schistosoma haematobium-exposed participants was cultured with schistosome egg, adult worm, and cercaria antigens pre- and 6 weeks post-praziquantel treatment. Innate inflammatory (tumor necrosis factor α [TNF-α], interleukin(IL-)-6, IL-8), Th1 (interferon γ [IFN-γ], IL-2, IL-12p70), Th2 (IL-4, IL-5, IL-13), Th17 (IL-17A, IL-21, IL-23p19), and regulatory (IL-10) cytokines were quantified via enzyme-linked immunosorbent assay. Cytokine data was integrated using nonmetric multidimensional scaling and factor analysis.Results. Egg-specific cytokine phenotypes became more proinflammatory post-treatment due to increased TNF-α, IL-6, IL-8, IFN-γ, IL-12p70, and IL-23 levels. Post-treatment cercariae-specific responses were also more proinflammatory reflecting elevated IL-8. In contrast, post-treatment adult worm-specific responses were less inflammatory, reflecting lower post-treatment IL-6. A combination of egg-induced IL-6, IL-12p70, IL-21, and IL-23 and adult worm-induced IL-5 and IL-21 post-treatment was associated with reduced reinfection risk 18 months later.Conclusions. Praziquantel treatment markedly alters polarization of schistosome-specific cytokine responses, and these changes, particularly in response to egg-stage parasites, may promote resistance to reinfection. |
Claire D Bourke; Norman Nausch; Nadine Rujeni; Laura J Appleby; Kate M Mitchell; Nicholas Midzi; Takafira Mduluza; Francisca Mutapi |
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Publication Detail:
Type: JOURNAL ARTICLE Date: 2012-10-8 |
Journal Detail:
Title: The Journal of infectious diseases Volume: - ISSN: 1537-6613 ISO Abbreviation: J. Infect. Dis. Publication Date: 2012 Oct |
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Created Date: 2012-10-9 Completed Date: - Revised Date: - |
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Nlm Unique ID: 0413675 Medline TA: J Infect Dis Country: - |
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Languages: ENG Pagination: - Citation Subset: - |
Institute of Immunology and Infection Research, School of Biological Sciences, University of Edinburgh, United Kingdom. |
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Journal Information Journal ID (nlm-ta): J Infect Dis Journal ID (iso-abbrev): J. Infect. Dis Journal ID (publisher-id): jid Journal ID (hwp): jinfdis ISSN: 0022-1899 ISSN: 1537-6613 Publisher: Oxford University Press |
Article Information Download PDF © The Author 2013. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. creative-commons: Received Day: 26 Month: 3 Year: 2012 Accepted Day: 6 Month: 6 Year: 2012 Print publication date: Day: 1 Month: 7 Year: 2013 Electronic publication date: Day: 8 Month: 10 Year: 2012 pmc-release publication date: Day: 8 Month: 10 Year: 2012 Volume: 208 Issue: 1 First Page: 159 Last Page: 169 PubMed Id: 23045617 ID: 3666130 DOI: 10.1093/infdis/jis524 Publisher Id: jis524 |
Integrated Analysis of Innate, Th1, Th2, Th17, and Regulatory Cytokines Identifies Changes in Immune Polarisation Following Treatment of Human SchistosomiasisAN1 | |
Claire D. Bourke1a | |
Norman Nausch1 | |
Nadine Rujeni1 | |
Laura J. Appleby1 | |
Kate M. Mitchell1b | |
Nicholas Midzi2 | |
Takafira Mduluza3 | |
Francisca Mutapi1 | |
1Institute of Immunology and Infection Research, School of Biological Sciences, University of Edinburgh, United Kingdom |
2National Institute of Health Research, Harare, Zimbabwe |
3Biochemistry Department, University of Zimbabwe, Harare |
Presented in part: August 2010 at the 12th International Congress of Parasitology (ICOPA), Melbourne, Australia, August 2010. Abstract ABSTS-DCLZC-E7Z2P-XZK5J; British Society of Immunology Congress, Liverpool, UK, December 2010. Abstract A-341-0003-00060. aPresent affiliation: Centre for Immunology and Infection, University of York, United Kingdom. bPresent affiliation: Faculty of Public Health and Policy, London School of Hygiene and Tropical Medicine, United Kingdom. Correspondence: Correspondence: Claire D. Bourke, PhD, Department of Biology (Area 5), University of York, York YO10 5DD, UK (firstname.lastname@example.com). |
Urogenital schistosomiasis is a debilitating disease caused by the parasitic trematode Schistosoma haematobium, which currently infects >100 million people [1]. Humans are exposed to infective larvae (cercariae), which invade via percutaneous penetration; adult male and female worms, which mate in the urogenital tract; and eggs, which are transmitted to the environment in urine. Repeated passage of eggs across the bladder and urogenital tract leads to characteristic immune-mediated morbidities [2, 3]. The World Health Organization advocates schistosomiasis control via treatment with the antihelminthic drug praziquantel (PZQ) [4] due to its efficacy at clearing infection [5, 6] and reducing the prevalence of schistosome-associated morbidities [7].
Praziquantel treatment also alters schistosome-specific immune responses, and removal of infection and/or exposure to antigens released from parasites damaged by treatment may influence responses to subsequent infection [8–10]. Studies showing a decline in the proportion of circulating T regulatory (Treg) cells [11] and increased effector responses to parasite [12, 13] and nonparasite antigens [14, 15] post-treatment indicate that immunoregulatory mechanisms established during infection are alleviated when infection is removed. Furthermore, PZQ-induced changes in antibody levels, CD23+ B cells [16], and cellular proliferation [13] have been associated with reduced Schistosoma mansoni reinfection rates, suggesting that treatment may have longer-term benefits in addition to transient infection clearance.
Taken together, these observations suggest that treatment alters not only the magnitude of the host immune response but also the phenotype of schistosome-specific responses. However, although previous human studies have inferred relative changes in the phenotype of the schistosome-specific cytokine response from changes in individual responses [12, 17], this hypothesis has not been directly assessed. Importantly, experimental models of helminth infection suggest that innate cells and CD4+ T helper (Th) cells (Th1, Th2, Treg, and Th17) cross-regulate one another during infection [18, 19], and the interaction between these responses rather than levels of individual cytokines alone may influence resistance to reinfection [20]. No studies to date have investigated schistosome-specific, Th17-associated cytokine responses in humans despite the role of Th17 in schistosome egg-mediated immunopathologies in mice [21–23] and detectable levels of Th17-type cytokines in plasma samples from S. haematobium–exposed humans [24].
This study addresses 4 key hypotheses relating to the effect of PZQ on schistosome-specific immune responses: (1) treatment influences levels of schistosome-specific, Th17-associated cytokines in addition to the previously assayed cytokines; (2) the cytokine environment (comprising markers associated with innate inflammatory, Th1, Th2, Th17, and regulatory responses) of treated individuals is fundamentally altered relative to pretreatment responses; (3) the effects of treatment on schistosome-specific cytokines differ between responses to cercariae, adult worms and eggs; and (4) cytokine responses elicited 6 weeks post-treatment influence reinfection rates. Our results yield insights into the immunological effects of antihelminthic treatment and how these relate to parasite life history and the development of protective immunity.
Ethical approval was granted by the Medical Research Council of Zimbabwe and the University of Zimbabwe's Institutional Review Board. Local permission was granted by the Provincial Medical Director. Community members were informed of the study aims and procedures in their local language (Shona), and compliant participants provided written consent or assent from a parent/guardian if aged <18 years.
Participants were recruited from Murehwa District an S. haematobium endemic region of Zimbabwe, which is part of ongoing immunoepidemiological studies [14, 25–27]. Recruitment was school-based, and non–school going community members were invited to attend via community meetings. After pretreatment sampling for parasitology and cytokine assays, participants were treated with a single dose of PZQ (40 mg/kg body weight). Blood samples for follow-up cytokine assays were collected 6 weeks post-treatment, and parasitological samples were collected 6 weeks, 6 months, and 18 months post-treatment. To be included in the study, participants had to meet the following 6 criteria: (1) life-long residents of the area; (2) provided at least 2 urine and 2 stool samples on consecutive days for parasitological analysis pretreatment and 6 weeks and 18 months post-treatment; (3) negative for S. mansoni, soil-transmitted helminths, malaria, and human immunodeficiency virus (HIV) at all 3 time points; (4) not previously treated with antihelminthic drugs; (5) received PZQ treatment after baseline sampling and cured of S. haematobium infection (ie, no eggs detectable) 6 weeks post-treatment; and (6) provided sufficient blood for cultures and all cytokine assays before treatment and 6 weeks post-treatment. No cases were excluded for parasite coinfection; 6 HIV-positive cases were excluded, 7 participants refused treatment, and 23 participants provided insufficient sample volume. In accordance with the Medical Research Council of Zimbabwe ethical guidelines on blood volumes that can be collected from young children, children aged <5 years were excluded. Based on these criteria, a total of 72 people were included in this longitudinal study. High community-wide infection levels precluded inclusion of an untreated control group.
Schistosoma haematobium, S. mansoni, and soil-transmitted helminth eggs were quantified via microscopic analysis of urine and stool samples using standard urine filtration and Kato–Katz protocols respectively [28, 29]. A minimum of 2 samples per participant collected on consecutive days and 2 slides per stool sample were analyzed to quantify mean infection intensity before treatment and 6 weeks and 18 months post-treatment. Some participants also provided parasitological samples 6 months post-treatment, and these were used to confirm reinfection status. Plasmodium spp. were identified via microscopic examination of blood smears and confirmed with a Paracheck rapid test (Orchid Biomedical Systems). HIV was detected using DoubleCheckGold HIV1&2 Whole Blood test (Orgenics), and positive cases were confirmed using Determine HIV1/2 Ag/Ab Combo (InvernessMedical).
Schistosoma haematobium cercariae antigen preparation (CAP), whole adult worm homogenate (WWH), and soluble egg antigen (SEA) were obtained from the Theodor Bilharz Institute, Giza, Egypt. Antigens were confirmed endotoxin-free using previously described protocols [30] (Supplementary Figure 1).
Venous blood samples were collected by trained nurses, diluted 1:3 in culture medium (Roswell Park Memorial Institute medium supplemented with 2 mM L-glutamine and 100 U penicillin/streptomycin; all Lonza), and cultured in duplicate wells coated with 10 µg/mL CAP, WWH, and SEA for 48 hours at 37°C in Anaerogen Compact anaerobic atmosphere generation pouches (OXOID). Unstimulated cultures (antigen-free media) were conducted in parallel. Fewer CAP stimulations were conducted (n = 21) due to sample limitations. Cell-free culture supernatants were frozen and assayed within 12 months.
Interferon γ (IFN-γ), tumor necrosis factor α (TNF-α), interleukin 2 (IL-2,) interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 8 (IL-8), interleukin 10 (IL-10), interleukin 12p70 (IL-12p70), interleukin 13 (IL-13), and interleukin 21 (IL-21) (BD Biosciences) and interleukin 17A (IL-17A) and interleukin 23p19 (IL-23p19) (eBiosciences) were assayed in culture supernatants via enzyme-linked immunosorbent assay. Ninety-six well plates were coated with 1 µg/mL or 2 µg/mL (TNF-α and IL-23 only) capture antibody in serum-free phosphate-buffered saline. Plates were washed 3 times and blocked for 2 hours with phosphate-buffered saline/2% bovine serum albumin (Alpha Diagnostic International). Recombinant cytokine standard dilutions were prepared from starting concentrations of 1 ng/mL (IL-17A), 2 ng/mL (IL-23p19 and IL-4), 5 ng/mL (IFN-γ, TNF-α, IL-5, IL-6, IL-8, and IL-10), 10 ng/mL (IL-13), 20 ng/mL (IL-2 and IL-12p70), and 40 ng/mL (IL-21). Supernatant samples were added to duplicate wells and incubated overnight at 4°C.
After 3 washes, 0.5 µg/mL (IFN-γ, TNF-α, IL-2, IL-5, IL-6, IL-8, IL-12p70, and IL-23) or 1 µg/mL (IL-4, IL-10, IL-13, IL-17A, and IL-21) biotinylated detection antibody was added for 2 hour at 37°C. After 4 washes, 1:6000 streptavidin–horseradish peroxidase (GE Healthcare) was added for 2 hour at 37°C. Plates were washed 4 times and incubated with 3,3′,5,5′-tetramethylbenzidine (TMB)-based streptavidin–horseradish peroxidase substrate solution (Sigma-Aldrich). Assays were developed for 1.5 minutes (IL-8 only) or 5 minutes, stopped with 25% hydrochloric acid, and read at 450 nm using SoftmaxPro spectrophotometer software (Molecular Devices).
Before analysis, mean cytokine concentrations in unstimulated wells were subtracted from those of the corresponding antigen-stimulated cultures for each participant to give antigen-specific cytokine levels.
Cytokine responses to CAP, WWH, and SEA pretreatment and 6 weeks post-treatment were square-root(x + 1) transformed to meet parametric assumptions and compared using repeated measures analysis of variance conducted using IBM SPSS Statistics v.19 software. Pair-wise comparisons were made using Fisher's least significant difference test [31]. Because multiple comparisons may give rise to false positive results, P values from analysis of variance were adjusted using the sequential Bonferroni method [32]. Raw P values are reported, and those significant postcorrection are considered highly significant.
In addition to 1-way analysis of cytokine levels, a data reduction method—multidimensional scaling—was used to visualize the multiple cytokine responses of each participant in a 2-dimensional space. The nonparametric version of this approach, nonmetric multidimensional scaling (NMS) [33], was used because combined data did not meet parametric assumptions. This approach was used to account for multicolinearity and multiplicity between the 13 cytokines and to identify how patterns of cytokine responses (an indicator of cellular immune phenotype) were affected by treatment. A single score was first assigned to each participant corresponding to their combination of CAP-, WWH-, or SEA-specific cytokine responses ranked relative to all other participants before treatment and 6 weeks post-treatment. Scores were then plotted via NMS ordination to give a visual representation of relative similarity and/or dissimilarity in participant cytokine responses [33]. Hypothesis testing comparing pretreatment and post-treatment NMS scores was conducted using the nonparametric multiple response permutation procedure (MRPP) [33]. Pearson's correlation between the original cytokine levels and NMS scores identified responses that varied most between participants. Nonmetric multidimensional scaling and MRPP were implemented using PC-ORD software as described in the Supplementary information.
Factor analysis was used to reduce all square-root(x + 1)–transformed, 6 week post-treatment SEA- and WWH-specific cytokine responses into a smaller number of variables (principal components [PCs]) reflecting cytokine profiles [31]. Only PCs that accounted for a greater than average proportion of variance in the original cytokine data (eigenvalue > 1) and correlated with at least 2 of the original cytokine variables with factor loadings ≥0.5 or ≤ −0.5 were included in subsequent analyses [31]. Binary logistic regression was conducted with sex, age, pretreatment infection intensity, and all SEA- and WWH-specific cytokine profiles (6 week post-treatment PCs) included simultaneously in the model as predictors of reinfection status (reinfected: ≥1 S. haematobium egg detected between 6 and 18 months post-treatment; uninfected: no S. haematobium eggs detected in post-treatment samples). The Log likelihood ratio test statistic (D) and associated P value were calculated for each predictor from the difference between the Log likelihood model fit for the full model to that where the predictor was not included.
Pretreatment prevalence of S. haematobium infection within the cohort was 58.3% with a mean infection intensity of 30.3 eggs/10 mL urine (standard error of the mean [SEM], 9.82; range, 0–481 eggs/10 mL urine). Participants had a mean age of 9.3 years (SEM, 0.31; range: 5–17 years); 40 were males, and 32 were females. Demographic and infection characteristics of the cohort are representative of the whole community [14].
Mean cytokine concentrations showed distinct post-treatment dynamics according to antigen stimulation (Figure 1 and Table 1). Concentrations of all SEA-specific responses, with the exception of IL-5, were higher than pretreatment levels, and this increase was statistically significant for innate inflammatory (TNF-α, IL-6, and IL-8), Th1 (IFN-γ, IL-2, and IL-12p70), and Th17 (IL-21 and IL-23p19) cytokines. Cercariae antigen preparation –specific IL-8, IL-10, and IL-13 responses were significantly higher 6 weeks post-treatment, but CAP-specific IL-23p19 levels were significantly lower 6 weeks post-treatment. Whole adult worm homogenate –specific TNF-α and IL-6 were significantly lower post-treatment than pretreatment. Interleukin 5 responses to all antigens were lower post-treatment than pretreatment, although this trend was not statistically significant. IL-21 responses to all schistosome antigens were higher post-treatment than pretreatment, but only significantly so for SEA and WWH. Changes in SEA-specific TNF-α, IL-6, IL-8, IFN-γ, IL-12p70, IL-21, and IL-23p19 and CAP-specific IL-8 were highly significant (ie, remained significant after Bonferroni correction).
Consistent with systemic effects of treatment, levels of spontaneously produced IL-12p70 and IL-21 increased and IL-2, IL-5, and IL-8 responses decreased in unstimulated cultures post-treatment (Supplementary Figure 2).
Having identified differences between the levels of individual cytokines following treatment, the implication of these changes for polarization of whole blood cytokine responses was investigated via NMS. Participant scores are plotted according to similarity and/or dissimilarity between cytokine profiles pre- and post-treatment (Figure 2). The individual cytokines that varied most between participants are indicated on spatial axes (correlation coefficients in Supplementary Table 1).
Pre- and post-treatment responses to CAP varied predominantly according to IL-6 (axis 1) and IL-8 (axis 2). Distinct groupings along axis 2 (Figure 2A) reflect the significant difference between pre- and post-treatment cytokine polarization (T, −17.25; P < .001; A, 0.199) and particularly the highly significant increase in cercariae-specific IL-8 post-treatment (Table 1).
Variation between pre- and post-treatment WWH-specific cytokine responses (Figure 2B) was mainly due to IL-6 (axis 1), reflecting the post-treatment decline in WWH-specific innate inflammatory cytokines (Figure 1). No cytokines were strongly associated with axis 2, and there was considerable overlap between pre- and post-treatment responses, suggesting that pre- and post-treatment cytokine profiles were similar, consistent with the weakly significant differences in individual WWH-specific cytokines (Table 1). Although MRPP indicated that WWH-specific post-treatment cytokine profiles changed relative to baseline (T, −9.37; P < .001), the effect size of the difference was small (A, 0.029).
Soluble egg antigen–specific cytokine responses exhibited a clear shift post-treatment (Figure 2C) according to a combination of TNF-α, IL-6, IFN-γ, IL-12p70, and IL-23p19 (axis 1) and TNF-α, IL-6, IL-8, IL-12p70, and IL-23p19 (axis 2) and significantly differed relative to pretreatment responses (T, −72.67; P < .001; A, 0.391). The distinct clustering and low overlap of pre- and post-treatment SEA-specific cytokine responses indicate that this post-treatment shift was evident in nearly all participants. The loading of IL-8 onto axis 2 but not axis 1 and IFN-γ onto axis 1 but not axis 2 indicates that participants differed in their IL-8 and IFN-γ responses in addition to the more uniform upregulation of other proinflammatory cytokines.
Spontaneous cytokine production was also affected by treatment predominantly due to variation in IL-6 and IL-8 (axis 1) and IL-21 (axis 2) after treatment relative to baseline (Figure 2D). Pre- and post-treatment cytokine responses in unstimulated cultures differed significantly (T, −2.804; P = .02), although the effect size of this difference was low (A, 0.008).
Of the study participants, 18 were reinfected within 18 months of treatment. Table 2 summarizes the characteristics of the reinfected participants relative to those that remained uninfected. Having identified reinfection within the cohort, we sought to investigate whether the risk of reinfection was related to the cytokine profiles mounted in response to S. haematobium adult worm and egg antigens 6 weeks post-treatment.
Factor analysis identified 4 profiles of 6 weeks post-treatment cytokine responses to WWH and SEA (Table 3). Principle component 1, which accounted for the most variation between cytokine profiles (15.6%), reflects regulatory and Th2-polarized cytokine responses due to its positive loading with SEA- and WWH-specific IL-10 and IL-13 and negative loading with SEA-specific TNF-α, IL-6 and IL-23. Principle component 2 reflects a combination of SEA-specific proinflammatory and WWH-specific Th2- and Th17-polarized responses. Principle component 3 reflects proinflammatory cytokine responses to WWH- (IL-6, IL-8, and IL-23p19) and SEA-specific IL-17A and was negatively loaded with Th2 cytokines. Principle component 4 comprised SEA-specific TNF-α and IL-13, both of which are associated with schistosome-associated immunopathology [34, 35].
Binary logistic regression analysis showed that participant age, sex, and pretreatment infection intensity were not significant predictors of reinfection status, but high PC2 factor scores (reflecting a combination of SEA-specific proinflammatory [IL-6, IL-12p70, IL-21, and IL-23p19] and WWH-specific Th2- (IL-5) and Th17-associated [IL-21] responses) were significantly associated with a lower risk of reinfection (Table 4). None of the other post-treatment cytokine profiles were significant predictors of reinfection status (Table 4).
Praziquantel treatment is known to affect levels of individual schistosome-specific immune responses and may do so directly by killing parasites and exposing their antigens to immune recognition [36, 37] and/or indirectly by removing immunosuppressive mechanisms mediated by live parasites [11]. The current study shows that the effects of treatment on schistosome-specific cytokine responses are dependent both on parasite life-cycle stage and the cellular phenotype with which they are associated. In particular, cercariae- and egg-specific, but not adult worm–specific, cytokine responses become polarized toward a more proinflammatory phenotype after treatment than that observed prior to treatment. Interestingly, participants who became reinfected within 18 months of curative treatment had lower PC scores for a combination of egg-specific proinflammatory and adult worm–specific IL-5 and IL-21 than their uninfected counterparts, suggesting that the boost in proinflammatory responses to egg antigens after treatment may promote resistance to reinfection. Importantly, quantification of innate-, Th1-, Th2-, Treg- and Th17-associated cytokine families allowed a more comprehensive analysis of how treatment affects schistosome-specific cytokine profiles than those described in previous studies. Use of a multivariate statistical approach also facilitated meaningful interpretation of highly complex responses, which are known to interact and cross-regulate one another during helminth infections [18, 19].
As initially hypothesized, levels of Th17-associated cytokines (IL-21 and IL-23) changed markedly 6 weeks post-treatment; however, the 2 cytokines had distinct antigen-specific dynamics. Soluble egg antigen–specific IL-21 and IL-23 increased in the context of a range of other proinflammatory cytokines, cercariae-specific IL-21 was unchanged, cercariae-specific IL-23 declined, and adult worm-induced IL-21, but not IL-23, increased post-treatment. Interestingly, despite being present at detectable levels in response to all antigens, IL-17A levels did not differ post-treatment, suggesting that IL-21 and IL-23 may have IL-17A– (or indeed Th17-) independent roles despite their shared association with the Th17 lineage. For example, murine IL-21 has also been implicated in Th2 function and alternative activation of macrophages [38]. Furthermore, murine dendritic, cell-derived, schistosome-specific IL-23 promotes T cell IL-17 in vitro, and heightened IL-23 levels may also elicit tissue-derived proinflammatory cytokines in vivo [23]. Further studies are required to ascertain the cellular sources and functions of IL-17, IL-21, and IL-23 in human schistosomiasis.
Distinct post-treatment responses to schistosome life-cycle stages suggest that the exposure of stage-specific antigens differs following treatment and is evident despite the overlap in somatic antigens [39]. For example, the low cytokine responsiveness to WWH post-treatment may be due to rapid removal of adult worms from the circulation [40], whereas ongoing contact with cercariae-infected water [41] and prolonged excretion of eggs from host tissues [5] may have maintained exposure to these life-cycle stages during the 6 weeks following treatment. Thus, our results do not preclude the possibility that cellular immune responses to adult worms are "boosted" at earlier post-treatment timepoints, as observed in S. mansoni studies [17, 42], but indicate that these effects decline within 6 weeks without repeated treatment.
Egg- and cercariae-specific proinflammatory responses may also increase post-treatment due to disruption of immunosuppression mediated by live parasites during infection [43]. This hypothesis is supported by observations that circulating Treg numbers decrease postclearance of S. mansoni [11], suggesting that infection elevates peripheral immunoregulation. However, observed changes in SEA-specific cytokines were not due to changes in egg-specific IL-10, which was unaffected by treatment. Thus, treatment-dependent elevation of SEA-specific proinflammatory cytokines may reflect changes in alternative mechanisms (eg, Tregs [35, 44]) regulating SEA-specific responses during infection. Consistent with the latter hypothesis, levels of spontaneous IL-10 were lower post-treatment than pretreatment, and we have previously shown that serological responses to non-parasite antigens increase post-treatment in the same population [14]. Regulation of egg-specific responses prior to treatment may be particularly important due to their immunopathological potential. For example, IL-13 and TNF-α, which are associated with egg-induced immunopathology [34, 35] and Th1 cytokines (IFN-γ and IL-12p70), which tend to be lower in chronically infected than uninfected humans [45, 46], were significantly higher 6 weeks post-treatment. The elevated IL-21 and IL-23 responses to SEA post-treatment are also consistent with a "rebound" in potentially pathological responses after removal of infection because Th17-associated responses exacerbate egg-induced granulomas in experimental schistosomiasis [21, 22].
This study also tested the hypothesis that post-treatment cytokine profiles influence the acquisition of new schistosome infections. Whereas some human helminth studies have focused on how pretreatment immune responses relate to reinfection [12, 47], we focused on post-treatment responses because reinfection occurs in the context of this altered immune environment. High post-treatment PC2 scores (comprising SEA-specific proinflammatory cytokines and WWH-specific IL-5 and IL-21) were associated with a reduced risk of reinfection within 18 months of treatment, consistent with an immune-mediated component to protective immunity rather than age-related changes in water contacts or immune physiology alone [48]. Only 18 participants were reinfected within 18 months of treatment, and thus reinfection analysis may have lacked statistical power to detect other significant associations. Nonetheless, our data provide the first evidence that Th17-associated IL-23 and IL-21 may contribute to protection from schistosome infection in addition to their putative immunopathological effects [21], and thus our data advocate further study of these cytokines in human schistosomiasis. The association between adult worm-specific Th2 cytokines and resistance to reinfection has been noted previously [9, 13] but never assessed in the context of innate inflammatory, Th1, Th17, and regulatory cytokines which can cross-regulate Th2 responses [19].
Because eggs are only produced after an infection has already been established, immune responses targeting egg antigens alone are unlikely to directly reduce reinfection. What seems more likely is that post-treatment, egg-specific, proinflammatory cytokine levels correspond to responses that limit de novo invasion by cercariae or the fitness and/or fecundity of adult worms. For example, SEA contains a range of antigens that cross-react with CAP, WWH, and schistosomules [39], and therefore increased reactivity to egg-stage parasites may also promote anti-larval and anti–adult worm responses [49]. Consistent with our observations, individuals with low peripheral blood mononuclear cell proliferative responses to S. mansoni SEA (and CAP) were more likely to be reinfected independent of age, sex, and pretreatment infection intensity in an Egyptian cohort [50]. Interestingly, we show that CAP-specific IL-8, IL-10, and IL-13 increase post-treatment; however, the limited number of cases for which CAP stimulation was conducted meant their relationship with reinfection rates could not be assessed.
This study provides the most comprehensive analysis of schistosome-specific cytokine responses in human schistosomiasis to date and the first description of human Th17 cytokine responses to schistosome antigens. Our results demonstrate that a single dose of PZQ markedly alters the polarization of schistosome-specific cytokine responses and that these changes, particularly in response to egg-stage parasites, may contribute to the development of resistance to reinfection.
Supplementary materials are available at The Journal of Infectious Diseases online (http://jid.oxfordjournals.org/). Supplementary materials consist of data provided by the author that are published to benefit the reader. The posted materials are not copyedited. The contents of all supplementary data are the sole responsibility of the authors. Questions or messages regarding errors should be addressed to the author.
Supplementary DataAcknowledgments. This study would not have been possible without the participation of Magaya community and the students and teachers of Magaya primary and secondary schools. We also acknowledge the support of technical specialists from the National Institute of Health Research, Harare, and the University of Zimbabwe, Harare, for collection and analysis of all parasitological samples; the nursing staff of Murehwa District Hospital, Murehwa, who collected all blood samples and administered treatment; and Noah Paul and students from the University of Zimbabwe, who assisted with whole blood sample processing at the field site. We thank the director and staff of Murehwa District Hospital for hosting our research; Dr Margo Chase Topping, University of Edinburgh, Edinburgh, for her advice on NMS; and Dr Adrian Mountford for comments on the manuscript.
Financial support. This work was supported by the Carnegie Trust for the Universities of Scotland; Tenovus Scotland; the University of Edinburgh Moray Endowment Fund; the Thrasher Foundation, the World Health Organization (grant RPC264); and the Wellcome Trust (grant WT082028MA). C. D. B. was funded by a studentship from the Biotechnology and Biological Sciences Research Council and a travel grant from the British Society of Parasitology Garnham Expeditionary Fund. These funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript.
Potential conflicts of interest. All authors: No reported conflicts.
All authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed.
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12. | Joseph S,Jones FM,Walter K,et al. Increases in human T helper 2 cytokine responses to Schistosoma mansoni worm and worm-tegument antigens are induced by treatment with praziquantelJ Infect DisYear: 20041908354215272413 |
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15. | van den Biggelaar AHJ,Rodrigues LC,van Ree R,et al. Long-term treatment of intestinal helminths increases mite skin-test reactivity in Gabonese schoolchildrenJ Infect DisYear: 200418989290014976607 |
16. | Mwinzi PNM,Ganley-Leal L,Black CL,Evan Secor W,Karanja DMS,Colley DG. Circulating CD23+ B cell subset correlates with the development of resistance to Schistosoma mansoni reinfection in occupationally exposed adults who have undergone multiple treatmentsJ Infect DisYear: 2009199272919072134 |
17. | Fitzsimmons CM,Joseph S,Jones FM,et al. Chemotherapy for schistosomiasis in Ugandan fishermen: treatment can cause a rapid increase in interleukin-5 levels in plasma but decreased levels of eosinophilia and worm-specific immunoglobulin EInfect ImmunYear: 20047240233015213147 |
18. | Allen JE,Maizels RM. Diversity and dialogue in immunity to helminthsNat Rev ImmunolYear: 2011113758821610741 |
19. | Diaz A,Allen JE. Mapping immune response profiles: the emerging scenario from helminth immunologyEur J ImmunolYear: 20073733192618000958 |
20. | Wilson MS,Cheever AW,White SD,Thompson RW,Wynn TA. IL-10 blocks the development of resistance to re-infection with Schistosoma mansoniPlos PathogYear: 20117e100217121829367 |
21. | Rutitzky LI,Bazzone L,Shainheit MG,Joyce-Shaikh B,Cua DJ,Stadecker MJ. IL-23 is required for the development of severe egg-induced immunopathology in schistosomiasis and for lesional expression of IL-17J ImmunolYear: 200818024869518250458 |
22. | Rutitzky LI,Smith PM,Stadecker MJ. T-bet protects against exacerbation of schistosome egg-induced immunopathology by regulating Th17-mediated inflammationEur J ImmunolYear: 20093924708119714576 |
23. | Shainheit MG,Smith PM,Bazzone LE,Wang AC,Rutitzky LI,Stadecker MJ. Dendritic cell IL-23 and IL-1 production in response to schistosome eggs induces Th17 cells in a mouse strain prone to severe immunopathologyJ ImmunolYear: 200818185596719050275 |
24. | Milner T,Reilly L,Nausch N,et al. Circulating cytokine levels and antibody responses to human Schistosoma haematobium: IL-5 and IL-10 levels depend upon age and infection statusParasite ImmunolYear: 2010327102121039611 |
25. | Imai N,Rujeni N,Nausch N,et al. Exposure, infection, systemic cytokine levels and antibody responses in young children concurrently exposed to schistosomiasis and malariaParasitology FirstView,Year: 2011115 |
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27. | Rujeni N,Nausch N,Midzi N,Mduluza T,Taylor DW,Mutapi F. Schistosoma haematobium infection levels determine the effect of praziquantel treatment on anti-schistosome and anti-mite antibodiesParasite ImmunolYear: 2012343304022429049 |
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35. | Wamachi AN,Mayadev JS,Mungai PL,et al. Increased ratio of tumor necrosis factor-alpha to interleukin-10 production is associated with Schistosoma haematobium–induced urinary-tract morbidityJ Infect DisYear: 200419020203015529268 |
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40. | Hassan MM,Medhat A,Makhlouf MM,et al. Detection of circulating antigens in patients with active Schistosoma haematobium infectionAm J of Trop Med and HygYear: 1998592953019715950 |
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50. | Colley DG,Barsoum IS,Dahawi HSS,Gamil F,Habib M,Elalamy MA. Immune responses and immunoregulation in relation to human schistosomiasis in Egypt 0.3. Immunity and longitudinal-studies of in vitro responsiveness after treatmentTrans R Soc Trop Med HygYear: 19868095273111030 |
Schistosome-Specific Cytokine Responses Differ 6 Weeks After Treatment Relative to Pretreatment Levels
Phenotype | Cytokine | Antigen
F Valuea | P Value | F Valuea | P Value | F Valuea | P Value | ||
Innate inflammatory | TNF-α | 3.70 | 0.07 | 6.18 | 0.02 | 75.24b | <0.001b |
IL-6 | 0.82 | 0.38 | 7.78 | 0.01 | 208.47b | <0.001b | |
IL-8 | 64.92b | <0.001b | 2.38 | 0.13 | 90.49b | <0.001b | |
Th1 | IFN-γ | 3.53 | 0.07 | 0.95 | 0.33 | 28.70b | <0.001b |
IL-12p70 | 2.29 | 0.15 | 2.77 | 0.10 | 94.52b | <0.001b | |
Th2 | IL-4 | 0.76 | 0.39 | 2.70 | 0.10 | 1.34 | 0.25 |
Regulatory | IL-10 | 9.09 | 0.01 | 3.45 | 0.07 | 1.38 | 0.24 |
Th17 | IL-17A | 0.07 | 0.79 | 1.04 | 0.31 | 0.26 | 0.61 |
IL-21 | 4.31 | 0.05 | 7.32 | 0.01 | 8.88b | <0.001b | |
IL-23 | 5.01 | 0.04 | 1.34 | 0.25 | 190.15b | <0.001b | |
Results of repeated measures analysis of variance comparing mean concentrations (square root(x + 1) transformed) of cytokines present in whole blood cultures stimulated with cercariae (CAP), adult worm (WWH), and egg (SEA) antigens before treatment with those elicited 6 weeks after a single 40 mg/kg body weight dose of praziquantel (CAP, n = 21; WWH, SEA, and unstimulated, n = 72). Significant differences (P < .05) are highlighted in bold.
Abbreviations: IFN-γ, interferon γ; IL-2, interleukin 2; IL-4, interleukin 4; IL-5, interleukin 5; IL-6, interleukin 6; IL-8, interleukin 8; IL-10, interleukin 10; IL-12p70; interleukin 12p70; IL-13, interleukin 13; IL-17A, interleukin 17A; IL-21, interleukin 21; IL23, interleukin 23; TNF-α, tumor necrosis factor α.
a Error degrees of freedom: 1, 20 (CAP); 1, 71 (WWH and SEA).
b Differences that are significant after Bonferroni adjustment for multiple comparisons.
Demographic and Schistosoma haematobium Infection Characteristics of the Study Cohort 18 Months After Praziquantel Treatment
Reinfection status
Characteristics | Uninfected | Reinfecteda |
No. | 54 | 18 |
Mean age (SEM) | 9.2 (0.37) | 9.3 (0.53) |
Males, females | 27, 27 | 13, 5 |
Mean pretreatment infection intensitya | 28.6 (11.0) | 35.6 (21.9) |
Pretreatment infection rangeb | 0–481 | 0–395 |
Pretreatment infection prevalence | 57.4% | 61.1% |
Mean post-treatment infection intensityb (SEM) | 0 | 10.2 (4.5) |
Post-treatment infection rangeb (SEM) | 0 | 0–65 |
Abbreviation: SEM, standard error of the mean.
a Reinfection classified as presence of ≥1 S. haematobium egg in ≥1 urine sample between 6 months and 18 months post-treatment.
b Infection intensity quantified as mean S. haematobium egg counts/10 mL urine.
Adult Worm and Egg-Specific Cytokine Profiles 6 Weeks After Treatment
Phenotype | Cytokine | Antigen | Principal Component
1 | 2 | 3 | 4 | |||
Th2/Regulatory | SEA Proinflammatory/WWH Th2/Th17 | WWH Proinflammatory | SEA Proinflammatory/Th2 | |||
Innate inflammatory | TNF-α | SEA | −0.5 | 0.3 | 0.1 | 0.5 |
Th1 | IFN-γ | SEA | −0.4 | 0.4 | 0.1 | 0.4 |
IL-12p70 | SEA | −0.4 | 0.5 | 0.1 | 0.4 | |
Th2 | IL-4 | SEA | 0.2 | 0.4 | −0.5 | −0.2 |
Regulatory | IL-10 | SEA | 0.7 | 0.1 | 0.2 | 0.4 |
Th17 | IL-17A | SEA | 0.2 | 0.4 | 0.5 | −0.5 |
% Total variance | 15.6 | 12.3 | 15.4 | 12.5 |
Antigen-specific cytokine factor loadings for principal components (PCs) 1–4 extracted by factor analysis of all Schistosoma haematobium whole worm homogenate (WWH)– and soluble egg antigen (SEA)–specific cytokine responses 6 weeks after praziquantel treatment (n = 72). Cytokines with factor loadings ≥0.5 or≤−0.5 for an extracted PC are highlighted in bold. The cellular immune phenotype with which the cytokines are associated is given for each PC. The percentage of total variance in cytokine responses accounted for by each PC is given at the bottom of the relevant column.
Abbreviations: IFN-γ, interferon γ; IL-2, interleukin 2; IL-4, interleukin 4; IL-5, interleukin 5; IL-6, interleukin 6; IL-8, interleukin 8; IL-10, interleukin 10; IL-12p70; interleukin 12p70; IL-13, interleukin 13; IL-17A, interleukin 17A; IL-21, interleukin 21; IL23, interleukin 23; TNF- α, tumor necrosis factor α.
Adult Worm– and Egg-Specific Post-Treatment Cytokine Responses Influence the Risk of Reinfection 18 Months After Curative Treatment
Predictor | D | P Value | Odds Ratio | 95% CI |
Sex | 2.54 | .11 | 2.65 | .77–9.10 |
Age in years | 0.06 | .81 | 0.97 | .76–1.24 |
Pretreatment infection intensity | 0.31 | .58 | 1.00 | 1.00–1.01 |
Th2 (PC1) | 2.36 | .63 | 1.56 | .88–2.77 |
SEA proinflammatory/WWH Th2 (PC2) | 5.67 | .02 | 0.48 | .25–.93 |
WWH proinflammatory (PC3) | 0.62 | .43 | 0.75 | .35–1.60 |
SEA proinflammatory/Th2 (PC4) | 0.35 | .56 | 0.81 | .41–1.61 |
Results of adjusted binary logistic regression analysis (n = 72) with reinfection status (uninfected or reinfected) as the dependent variable and sex, age, pretreatment infection intensity (mean egg count/10 mL urine), and factor scores for whole blood cytokine profiles 6 weeks after treatment identified via factor analysis (Table 2) included as linear predictors. Analysis of each predictor was conducted adjusting for all other predictors. Significant predictors (P < .05) of reinfection status are highlighted in bold. Hosmer–Lemeshow statistic (full model): 7.794; P = .45.
Abbreviations: CI, confidence interval; D, Log likelihood ratio statistic (difference between model excluding the predictor and the full model); PC, principle component.
Article Categories:
Keywords: Human, helminth, cytokine, schistosomiasis, immune response, praziquantel. |
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MEXICO CITY -- A former Texas high school football player and petty street dealer who allegedly rose to become one of Mexico's most savage assassins says he personally knew the country's top drug lords and shipped cocaine from Colombia through Panama.
In a video released by Mexico's federal police, Edgar Valdez Villarreal, known as "the Barbie" for his fair complexion and green eyes, also told his interrogators that he transported cash hidden in trailers.
With his arrest Monday, Valdez became the third major drug lord brought down by Mexico in less than a year. The 37-year-old Valdez faces charges in three U.S. states for trucking in tons of cocaine.
"I have work ... investments, there in Colombia," he said, laughing, on the tape that was broadcast late Tuesday and provided to news organizations, including The Associated Press.
When asked if he worked in drugs, he replied yes.
U.S. prosecutors say Valdez has been the source of tons of cocaine smuggled into the United States.
The arrest was portrayed by the Mexican and U.S. governments as a victory for President Felipe Calderon, who is trying to recover public support for his war on organized crime in the face of escalating violence.
Authorities also said Valdez could provide intelligence on other top traffickers, including Sinaloa chief Joaquin "El Chapo" Guzman, Mexico's most-wanted drug lord.
Valdez told interrogators that he knew the principal leaders of the drug cartels, such as Guzman, the brothers Arturo and Hector Beltran Leyva, Ismael "El Mayo" Zambada and Jose Gerardo "El Indio" Alvarez, whom he called his friend.
During the video, Valdez sometimes looked up and laughed. He constantly wiped sweat with a tissue or shirt sleeve, though he did not look nervous.
Local media also showed a video from inside the three-level residence where he was arrested, including paintings of religious subjects, horses and flowers, Gucci and Cartier boxes, big-screen TVs, a pool table and a bar.
Mexican police said they chased Valdez across five Mexican states for a year, a pursuit that intensified in recent months as they raided home after home owned by the drug lord, missing him but nabbing several of his allies.
His arrest also yielded computers, telephones and other equipment authorities said would likely provide more information about his group.
Valdez's presentation before the media Tuesday coincided with an announcement that Colombian authorities had detained 11 people allegedly linked to the Mexican kingpin in that South American cocaine-producing country. Mexican Federal Police Commissioner Facundo Rosas said the arrests were likely related, with Colombian authorities taking advantage of a break in his organization.
As a U.S. citizen living illegally in Mexico, Valdez could be deported to the United States if Mexico agrees, or he could face prosecution in Mexico for drug-related crimes. Mexican authorities say he could be responsible for dozens of murders.
Born in the border city of Laredo, Texas, Valdez grew up in a middle-class subdivision popular with Border Patrol agents, police officers and firefighters. His father was a nightclub and bar owner.
The former Laredo United High School linebacker became a small-time street dealer as a teen, before rising to become the head of a group of assassins for Mexico's notorious Beltran Leyva gang, allied with the powerful Sinaloa cartel, according to U.S. and Mexican officials.
After leaving Texas for Mexico, Valdez quickly rose through the ranks of the Beltran Leyva cartel, police say. He was anointed head of Acapulco operations by cartel leader Arturo Beltran Leyva after serving as the drug kingpin's top bodyguard, Rosas said.
In Mexico, Valdez built up a life of luxury, with homes in the most expensive neighborhoods of Mexico City. Valdez said in the tape that he commissioned a movie on his life and invested $200,000 in it, but it was never released.
"He was flamboyant, he felt like he was untouchable," said a senior U.S. law enforcement official, who wasn't authorized to speak on the record.
That life started to crumble as Mexican law enforcement took on the Beltran Leyva gang. Two years ago, a widespread corruption probe toppled the cartel's top government protectors, including Mexico's former drug czar.
The biggest coup came in December, when Mexican marines killed cartel lord Arturo Beltran Leyva during a gunbattle in Cuernavaca.
That unleashed a gruesome fight between Valdez and Beltran Leyva's brother, Hector, the only one of the cartel's founders who was still at large. Decapitated and dismembered bodies littered the streets of Cuernavaca and Acapulco -- and often hung from bridges -- along with messages threatening one of the two feuding factions.
Valdez confirmed on the tape that he berated one accomplice for shooting Paraguayan soccer player Salvador Cabanas during an argument at a Mexico City bar in January, an attack that led to the arrest of a minor henchman, who later told police about Valdez's fury over the incident.
Valdez said he hid the suspect, Jose Jorge Balderas, also known as "JJ", who is still at large.
More of his allies fell in a series of raids and shootouts in Mexico City, Acapulco and other towns. Fifteen Valdez henchmen were killed in a battle with soldiers in June in Taxco, a mountain hamlet outside of Mexico City.
The pursuit intensified six weeks ago when Mexican security officials began getting tips on Valdez's whereabouts and approached U.S. agents for help, according to U.S. law enforcement officials in Washington, who spoke on condition of anonymity because of the sensitivity surrounding the arrest. U.S. intelligence helped pinpoint his location Monday.
An elite, U.S.-trained Mexican federal police squad arrested Valdez and four accomplices at the entrance of a ranch outside Mexico City.
"We were on his heels for the last six weeks, receiving tips, but Mexican law enforcement would show up and they would miss him," one U.S. official said. "He was feeling the heat of Mexican law enforcement."


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According to TMZ, Rodney King, who became a household name after his videotaped beating by Los Angeles police officers sparked a chain of events that lead to the deadly LA riots of 1992, has died at 47 at his home in Rialito, Calif. The website reports that law enforcement sources say he was found in the bottom of his pool. There has been no official confirmation of his death.
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Wednesday, August 06, 2008
4 comments:
- UrbanTulip said...
Μα που τα βρίσκεις όλα αυτά καλέ;
- kotosalata said...
Τα μοιράζανε στον Ηλεκτρικό της Κηφισιάς παρακαλώ!
- καιρος said...
α καλα εχει ξεφυγει ο κοσμος
καιρος - 6/21/2009 1:21 AM
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2013 Touch Screen Hang.
This was our first weekend out on the new X10 (I picked her up Friday). First thing this morning I set the profile to 'Surf Left' , activated the profile and about halfway thru filling the ballast the 7 inch (HV700) screen froze. The ballast pumps would not shut off (a good bit of water came out of the overflow before they finally stopped) and the cruise control was stuck on 10 mph.
Does anyone know of a key combination to force a reboot of the screen? I ended up pulling connectors to cut the power and force a reboot. It worked fine for the rest of the day today. Kind of disturbing so much key functionality is tied to this unit and there is no visible hard reset button anywhere and there are not manual overrides for all the critical components like the cruise. |
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03.24.17 – 04.01.17
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News Archive International
Monday, May 23, 2011
After the only light wind race on Saturday, the breeze filled in more and more on Sunday allowing to sail another four heats with up to 15 kn at the end. Thanks to two bullets in the morning, 'Foncia' with Xavier Revil FRA (standing in for Michel Desjoyeaux FRA) steering the multihull took the first place of the Realstone Cup, followed by the Geneva CER team with Jerôme Clerc SUI an rank 2 strongly coming back during the afternoon, extending the overall lead in the Vulcain Trophy 2011, the annual competition. 'Veltigroup' with Stève Ravussin SUI finished 3rd. The report and the interims rankings of the Vulcain-Trophy. The newcomer-team 'Artemis Racing' with Torbjörn Tornquist SWE and Paul Cayard USA is on the 4th place. Cayard USA, having gone thru many critical sailing situations in his round-the-buoys and offshore career, was impressed: 'I got my first taste of being scared in this boat.'
Décision 35 - Vulcain Trophy - Realstone Cup - Crans-sur-Nyon
Friday, 20. May 2011– Sunday, 22. May 2011
Friday, 20. May 2011– Sunday, 22. May 2011
» view all news for Regatta Décision 35 - Vulcain Trophy - Realstone Cup - Crans-sur-Nyon
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Olympic Classes - Training Camp Trophy - El Arenal ESP - Final results, Henken/Scutt ...
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Farr 40 - Sydney Open - Sydney AUS - Final results
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I loaded the money gem (Ruby 1.9.1), but it is not found. $sudo gem install money -v 2.1.4 Any ideas? When I hit my webapp in a browser, I get this in the tomcat logs. What is up with money module? --- /home/user/.rvm/rubies/ruby-1.9.1-p431/lib/ruby/site_ruby/1.9.1/rubygems/custom_require.rb:36:in `require': no such file to load -- money (LoadError) from /home/user/.rvm/rubies/ruby-1.9.1-p431/lib/ruby/site_ruby/1.9.1/rubygems/custom_require.rb:36:in `require' from /home/user/projects/access/mod/mod-common/trunk/kbr/web_reports/cgi/RC_Helper_Html.rb:7:in `<top (required)>' from /home/user/projects/access/mod/mod-common/trunk/kbr/web_reports/cgi/RC_Home.rb:2:in `require' from /home/user/projects/access/mod/mod-common/trunk/kbr/web_reports/cgi/RC_Home.rb:2:in `<main>' [Mon Jun 03 14:00:40 2013] [error] [client 127.0.0.1] Premature end of script headers: RC_Home.rb
on 2013-06-03 22:06
on 2013-06-05 02:40
J. V. <firstname.lastname@example.com> wrote: > I loaded the money gem (Ruby 1.9.1), but it is not found. > $sudo gem install money -v 2.1.4 > > Any ideas? Since you're using per-user rvm, DON'T sudo gem install. That uses the ruby that is available to root, which is likely NOT the ruby or gem space you want, since you are using per-user rvm. Seriously.
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For those who enjoy casting a skyward eye, August presents three opportunities to be swept up in astrological grandeur. The first occurs Aug. 10, when the full moon coincides with perigee, the closest point in its elliptical orbit around Earth.
I've been fortunate to spend a large part of my life living on islands that range from remote and minuscule to Manhattan. Many might say Manhattan doesn't count, but no one who has lived there would agree. It's true that the vertical nature of the city draws your eye up — you need to venture to the city's edges or heights to see the water that surrounds you. But there's no going anywhere without a bridge or a tunnel or a ferry.
Clambakes are fun, signature events of summer. But there's nothing like sitting around a simple beach fire with family or a few friends to make a summer evening really special.
There's something about staring into a driftwood fire, with everyone's faces alive in the light, that makes you sense your dusty ancestors.
Like many of us, I spend more and more of my life tethered to a bevy of electronic devices — smartphone, laptop, desktop, tablet. Am I ever without my phone?
I grew up with field guides for fish, shells, seabirds and the hundreds of other denizens of the shallows and the deep. As a boy, I not only wanted to know their names but also their habits and habitats, their food, range, migration and so on.
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Limited: Target another player: Look at his hand, except for one card of his choice. Lose 2 Honor.Limited: Destroy your performing Personality and target another player: He targets and bows one of his unbowed Personalities.
"My sources indictate that the Lion are weak toward the west, lord Junnosuke." - Hantei Naseru


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We ordered the Yamadaya Ramen with Tonkatsu Kotteri broth and the Spicy Ramen. Both were delicious. Both had a very deep, porky, rich milky broth. The Kotteri broth had extra savory depth from the black garlic oil as well as thick, melt in your mouth pieces of pork and back fat. It's definitely a dish for pork lovers. The "Yamadaya" style just added some more of the regular ramen toppings like seaweed, menma, and an extra egg to the dish, which was nice to balance out the richness. The Spicy Ramen actually had some decent heat, which is usually not the case with most spicy ramen out there, like at Santouka. The noodles were very thin, with pretty good texture, not too mushy. We switched dishes midway through the meal; I'm not sure that I could finish an entire bowl of one type of ramen as it is so rich and heavy, in slightly different ways. A little variety in broth definitely helps. You can tell how oily the broth is; both formed slight films on top after cooling down a bit. Overall, it is my new favorite ramen in the westside. I can't discern any MSG and I think the broth is better than Santouka and overall more interesting than Ramenya. It's definitely very heavy and very rich; it's been five hours since lunch and I'm still stuffed. I'll definitely be back as it was really good, but it's one of those places where I'll only go every once in awhile as it really is too much for regular consumption.
Yamadaya Ramen with Tonkatsu Kotteri broth |
Spicy Ramen (spicy #2 on a scale of 3) |
Pretty thin noodles |
11172 Washington Blvd.
Culver City, CA 90232
(310) 815-8776


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Reversed Button
sorry I had problems with my Internetbrowser.
My Question is the following.
If I used a wrong button, how could I make that reversed?
For example I tried to make the first tutorial, in which I must make a partition with "explode into 5 vertices by selection". I used the wrong command in Salome.
How can I make this reversed?
Is there a "reverse-Button" in Salome???
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Join OurStage to discover and listen to new music from great indie artists.
Get exposure building your fan base and selling music.
Find opportunities through music licensing and our competitions.
Artist Sign Up
I've been a guitar junkie since I was 11 and I've been composing with synthesizers since I was 14. I also gained an intense love for classical music when I was 17 and for this reason, began to learn how to play the piano.
I'm 27 today and still really love writing/playing/producing music.
My primary influences include: Vangelis, John Williams, Hans Zimmer, Tosin Abasi, Joe Satriani, Jean Michel Jarre, BT, Beethoven, J.S. Bach, and, most currently, Chopin.


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Kabocha Vanilla Chai Ice Cream
Author Notes: As an Ohio girl, I love frequenting Jeni's each time I'm in Columbus. Unfortunately, I've lived overseas for the past 7 years so these visits are infrequent (though I've been known to go twice in one day!). You can imagine my excitement when I heard Jeni had wrote a cookbook -- I promptly purchased the book. I've had fun creating her flavors at home and spreading Jeni's ice cream love around Japan!
Even though it's hot hot hot in Japan, I'm looking forward to cooler weather and autumn foods. For me, that means pumpkin. I thought adding the warming flavors of vanilla and chai would be a nice mix. I hope you agree!
Recipe is adapted from Jeni's ice cream base and various techniques from her cookbook. —Courtney Pollock
Food52 Review: WHO: Courtney Pollock is a new Food52 user -- this is her first recipe!
WHAT: A delicately spiced ice cream with Japanese kabocha squash, perfect for fall.
HOW: The milk is infused with chai flavors to add extra oomph to Jeni's classic base, and roasted, pureed kabocha squash adds body.
WHY WE LOVE IT: Like a more delicate pumpkin pie, this ice cream is perfect for the changing seasons. —The Editors
Makes approximately 1 quart
Roasted Kabocha Squash & Chai Milk Base
- 1 Kabocha squash
- 1 1-inch piece of fresh ginger, cut into thin rounds
- 1 teaspoon black peppercorns
- 5 whole cloves
- 3 cardamom pods
- 1 cinnamon stick
- 1 vanilla bean, split and scraped
- 3 cups whole milk
- 3 bags, Darjeeling tea
- For Kabocha Squash: Preheat oven to 400 degrees F. Cut the squash in half; remove seeds and membranes. Place cut side down on a baking sheet and roast for 40 minutes until soft. Let cool.
- Scoop the flesh into a food processor and puree until completely smooth. Measure out 1 cup of kabocha puree for the ice cream.
- For Chai Milk Base: Place next 5 ingredients (ginger through cinnamon stick) in a saucepan. Lightly crush with wooden spoon. Add vanilla bean (inside & bean) along with whole milk. Place over low heat and simmer for 20 minutes.
- Remove from heat and place in tea bags. Let steep for 10 minutes. Remove tea bags and strain milk to remove solids. Measure 2 cups of chai milk mixture for recipe.
Ice Cream (based on Jeni's ice cream base)
- 2 cups chai milk base
- 1 tablespoon plus 1 teaspoon cornstarch
- 1.5 ounces cream cheese, softened
- 1/8 teaspoon fine sea salt
- 1 1/4 cups heavy cream
- 2/3 cup sugar
- 2 tablespoons light corn syrup
- 1 cup kabocha puree
- 1/4 cup maple syrup
- Mix 2 tablespoons of chai milk base with cornstarch in a small bowl to make a smooth slurry.
- Whisk the cream cheese and salt in medium bowl until smooth. Add kabocha puree and maple syrup. Combine until smooth; set aside.
- Combine remaining chai milk base, cream, sugar and corn syrup in a 4-quart saucepan; bring to a rolling boil over medium-high heat and boil for 4 minutes. Remove from the heat and gradually whisk in the cornstarch slurry.
- Return the mixture to a boil over medium-high heat and cook, stirring until the mixture is slightly thickened, about 1 minute. Remove from heat.
- Slowly pour the hot cream into the kabocha squash mixture, whisking to combine.
- Pour the mixture into a 1-gallon freezer bag and submerge the sealed bag in an ice bath. Place ice bath in the refrigerator and leave for at least an hour, or preferably overnight.
- Pour the ice cream base into the frozen canister and spin until thick and creamy.
- Pack the ice cream into a storage container, press the sheet of parchment directly against the surface, and seal with an airtight lid. Freeze until firm, at least 4 hours.
- Your Best Ice Cream Contest Finalist!
More Great Recipes:
Ice Cream & Frozen Desserts|Ice Cream


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The Economic Approach to Human BehaviorSince his pioneering application of economic analysis to racial discrimination, Gary S. Becker has shown that an economic approach can provide a unified framework for understanding all human behavior. In a highly readable selection of essays Becker applies this approach to various aspects of human activity, including social interactions; crime and punishment; marriage, fertility, and the family; and "irrational" behavior. "Becker's highly regarded work in economics is most notable in the imaginative application of 'the economic approach' to a surprising breadth of human activity. Becker's essays over the years have inevitably inspired a surge of research activity in testimony to the richness of his insights into human activities lying 'outside' the traditionally conceived economic markets. Perhaps no economist in our time has contributed more to expanding the area of interest to economists than Becker, and a number of these thought-provoking essays are collected in this book."—Choice Gary Becker was awarded the Nobel Prize in Economic Science in 1992. |
レビュー - レビューを書く
The Economic Approach to Human Behavior | 3 |
Price and Prejudice | 15 |
Effective Discrimination | 17 |
Law and Politics | 31 |
Competition and Democracy | 33 |
Crime and Punishment An Economic Approach | 39 |
Time and Household Production | 87 |
A Theory of the Allocation of Time | 89 |
Irrational Behavior and Economic Theory | 153 |
Marriage Fertility and the Family | 169 |
An Economic Analysis of Fertility | 171 |
On the Interaction between the Quantity and Quality of Children | 195 |
A Theory of Marriage | 205 |
Social Interactions | 251 |
A Theory of Social Interactions | 253 |
Altruism Egoism and Genetic Fitness Economics and Sociobiology | 282 |
The Allocation of Time and Goods over Time | 115 |
On the New Theory of Consumer Behavior | 131 |
Irrational Behavior | 151 |
295 | |
310 | |
他の版 - すべて表示
allocation altruism amount analysis assortive mating assumed assumption average Becker behavior births budget Coase theorem commodities compensated constraint consumer consumption contraceptive correlation crime decisions decline demand curves depends determined discrimination discussion economic approach economists efficient egoist elasticity of demand equal equation example expenditures factor fertility firms forgone earnings full income gain from marriage greater household production function human capital implications important income effects income elasticity increase indifference curves inputs interaction irrational labor force less loss function marginal cost marginal utility married maximize measured money income Negroes number of children number of offenses opportunity set optimal sorting parents percentage persons polygyny positively related preferences punishment quantity rational real income reduce relative prices responses result rise sector shadow price social environment social income sociobiology spent substitution substitution effect tastes theorem theory tion total output traits transfers United utility function variables wage rates women


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"There has always been a thought that Australian players don't like it when other captains stand up to them. They like to make fun of a Mike Brearley or a Jeremy Coney, but put a Sourav Ganguly or an Arjuna Ranatunga in front of them - someone who stands up and mouths off -and they hate it." - Jarrod Kimber
I'm aware, but Spielberg's fingerprints were all over it. Granted that this was the intention, but still.
Super 8 was good enough to keep me entertained for the duration.
Can't wait to see Kate Beckensale in that outfit you mentioned Fusion.
If you'd watched the first, you'd have been silly to hope for anything above slight mirth.
Have seen it described by blokes forced to see it as Black Beauty goes to war. Basically only sappy, gullible women who've said anything good about it, and thats never beyond 'awww, so cute, so sad, awww, horsies'
Took me a while to warm to it, didn't straight away click with the characters, a by product of loving the Guinness version. Wasn't overly impressed with Oldman in it, think John Hurt would've been better as Smiley. Only ones I feel did a sterling job are Cumberbatch as Guillam and Toby Jones as Percy.
"All are lunatics, but he who can analyze his delusions is called a philosopher." - Ambrose Bierce
Langeveldt: I of course blame their parents.. and unchecked immigration!
GingerFurball: He's Austrian, they tend to produce the odd ****ed up individual
Burgey: Be careful dealing with neighbours whose cars don't have wheels but whose houses do.
Uppercut: Maybe I just need better strippers
Toby Jones is just great, think all the performances are excellent but his may be the best of the lot, still need to read the book.
Not too sure how I feel about TTSS. It was a good watch with great acting, but it's got nothing on the book.
Pretty much agree with cpr (as always).
I really enjoyed it, Oldman was immense in it.
I ended up getting dragged to watch The Girl With The Dragon Tattoo by the girlfriend, never got round to watching the original series of films but this had me glued from start to finish,well worth going to, very dialogue driven movie with a brilliant cast and the muisIcal score was brilliant. Friends say, for a change this version improved on the Swedish original, which I'm going to have to get round to watch soon.
Watched Dead Mans Shoes again last night, haven't seen it for a few years, what a ****ing film, highly recommend it to anyone who hasn't seen it.
Last edited by Jacknife; 23-01-2012 at 07:29 PM.
Paddy Considine Is close to a deity. still need to see Tyrannosaur, have only heard good things.
The problem is, when you've seen Alec Guinness do it, Oldmans got no chance....
I felt Oldman was too angsty, too driven, too forceful for Smiley.... His character's greatest strength was his silence, and his ability to draw your knowledge out of you, mainly through empathy and getting you to open up. His greatest moments were when he was just sat, thinking. Hence why the Karla meeting is such a key moment, the one time he shows weakness and opens himself to his only equal.
Certainly don't see the Oscar nomination in the performance. Guess its been a quiet year.
Cant remember if i said it, but watched The Guard the other night... Guess a sweary racist isn't the right character for an Oscar nod, but by god Gleeson was good. Also Mark Strong is guaranteed to appear in every British film nowadays, it seems. Not a bad thing mind.
Mark Strong is a great actor and I always enjoy his work.
One British actor whom I haven't seen in a recent film is Jeremy Northam, and he looks a lot like Mark Nicholas.
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Why Guitarists Spend Half Their time Tuning
And the Other Half Playing Out of Tune
A scholarly tome covering the physics of tuning, guitar adjustment and psycho-acoustics.
Printed by permission of Steve Mason, Luthier
science underlying this discussion more deeply, you can ask Steve Mason at: firstname.lastname@example.com.
FACT: C# AND Db ARE NOT THE SAME NOTE
In fact C# is slightly higher than Db, but they are played by the same key on the piano, and there is only one fret for them on each string on a guitar. What gives?
There is a mathematical relationship between every note in a scale. I won't bore you with all the math but the simplest relationships are: the octave is twice as many vibrations per second as the prime and the fifth is the prime plus one third of the difference between the prime and the octave. If you start on C and do all the math to calculate the vibrations per second for all the notes in a C scale and then calculate the vibrations per second for a D scale you will see that an E in a C scale is a slightly different note than an E in a D scale. This means that if your guitar is perfectly in tune to play a C chord and you want to play a perfect D chord you must raise your E and B strings a little. People apply "logic" to this situation and conclude that the E and B pegs are loose and need to be tightened or replaced or even soldered.
J.S. Bach in 1750 solved the problem with a system called "equal temperament", a system in which all the notes are slightly out of tune so that nothing is shockingly out of tune and you can play in any key with equal success. Pianos are tuned in equal temperament, and guitar fingerboards are fretted in equal temperament. Fiddlers (the good ones anyway) play in perfect pitches because they can micro adjust their finger positioning to play exactly the vibration per second required. The fixed frets of a guitar preclude this option.
A piano is a big clunky thing more closely related to a Chevy Mini-Van than to a violin. The tuning is expensive and takes a long time, so you tend to tune it and then play it as it sits. The guitar is more complicated: even though the frets are placed in equal temperament, guitar tuning pegs are easily accessible, and the strings can be "bent" with your fingers as you play.
Hence, the guitar player with a good ear is at constant war adjusting the string tension to approximate "just intonation" against the intentional misplacement of the frets. Very roughly, the first fret is 1/18 the distance from the nut to the saddle, the second fret is 1/18 the distance from the first fret to the saddle etc. In other words there is an equal distance between the notes (expressed logarithmically as the twelfth root of two). There is such a thing as a "just intonation fingerboard", the frets don't go straight across and are not evenly spaced. They play perfectly in tune, but they are only good for one key and are way out of tune in other keys.
The picture is further complicated by the physical makeup of your strings. As a string gets old it develops metal fatigue at some spots and not others, it gets a little dent everywhere it goes over a fret, the windings fill up with crud, the steel part of the string can rust and you can even wear it thin in spots. This uneven stress makes the string vibrate unevenly, so that the center point of the string is no longer the octave harmonic. You may have noticed that the first symptom that you need to change strings, just before they get "thumpy", is that you just can't get in tune.
And even fresh strings are imperfect. A string must be stretched to 80% of breaking tension so that the pull on its molecular bonds will make it act more like a fluid than like a rod. People using lots of different tunings will have noticed that a string tuned too low for its gauge won't "sing". A nice new string is great at producing primary vibrations but as the harmonics get higher and higher they are produced by smaller and smaller fractions of the string, and at some point the string section is so short that it acts more like a rod than like a fluid, and that pushes these high harmonics sharp.
The "action" of a guitar is the playability as governed by the straightness of the neck, the evenness of the fret tops, the roundness of the fret tops, the height of the nut and the height of the saddle, and the angle from the bridge to the pin holes and the nut to the peg barrels. This is all adjustable by adding material or taking material away. Guitars do not come adjusted from the factory. All new guitars must be adjusted (Martin makes no pretense about adjusting their new guitars; they don't even fit their bridge and butt pins). It takes this amalgamation of fine woods a while to get used to being a guitar, so you will probably have to readjust your action at six months and then yearly for the first five years. Every time the weather changes your guitar wood changes shape. If it bows backward it can come forward again, but if it bows forward it has 180 pounds of string tension holding it there. The normal motion is for the neck to bow up over time. It is then either a simple or difficult process to straighten it, depending on the brand of your guitar. After five years or so your guitar should stabilize to the point where you may go for two or three years between adjustments.
As you play, the high carbon steel strings wear down the nickel silver frets. This makes low spots on your favorite frets and turns the frets you rarely use into high frets. When you push a string down to a low fret it will buzz against the high fret in front of it. You can always miss that high fret by jacking your action up higher and higher. Every music store in the world knows how to shim your saddle to stop a buzz. This solves the problem, but makes your strings hard to push down. The proper solution is to let an experienced luthier "dress" your frets: file the fret tops so that all the fret tops are at the level of your lowest wear. This puts little flat spots on the tops of the frets that were too high. They are then rounded and polished. A good guitar in normal use can probably be adjusted 7 to 10 times before the frets are too low (worn about half way through) and must be replaced. However some very hard players wear through their frets in a few years.
So here's how this relates to the topic at hand: if your strings are too high you must "bend" them down to the frets. The extra stretch that it takes to get the string down to the fret tightens it in the same way as turning the tuning peg does. The string length from nut to saddle is designed (with more or less accuracy, depending on the guitar maker) to play in tune (equal temperament) at a certain string height. If you exceed that height the pitch will be sharp by the time the string contacts the fret.
Fret misplacement (actual misplacement, not to be confused with intentional misplacement) is very rare on modern guitars but saddle misplacement is common. In fact, saddle placement on vintage guitars seems quite random. If new strings do not yield a good match between your octave harmonic and your 12th fret go see your luthier. If you are using vastly different strings than the guitar manufactured intended, their bridge placement, though perfectly within their specs, will have to be changed for you.
Many people notice the tuning dilemma for the first time after they have shelled out the big bucks for a high quality guitar. A fine guitar has the ability to project, clearly and resonantly, the nuances of the string vibrations. Hence, you are more likely to hear the difference between C# and Db on your new Taylor, Collings, Santa Cruz, Martin, Larrivee, or Thompson than you were on your old junker. Also, on a fine guitar you don't notice that your strings are dead as quickly and you tend to not change them as often.
This is the study of how the human hearing system works. One major discovery of this field is that our hearing is often affected by things that we can't hear and we hear things that aren't really there. Another good reason to take up the violin is to hear a great demonstration of psycho acoustics: Play "Boil em Cabbage Down". As you play C# and E you will clearly hear an "undertone" of A, 2 octaves below. As you play the tune it sounds as if someone is playing bass with you. If you give the human ear the 1 and 3 it will interpolate the 5 of the chord when it is not being played.
And, perfect intonation does not necessarily please us. According to psycho acoustic research, we tend to like the high notes a little sharp and if any note is out of tune our first inclination is to "tune up". When players do the move from a D chord to a C chord, instead of lowering the E and B they tend to raise the bottom four strings. When they go back to D they raise the E and B and then when they switch to C they raise the bottom four again. Some of you old timers may remember jam sessions before the days of electronic tuners when the fixed pitch instruments would have to drop out by 9:00 and by 11:00 the guitars were tuned up to F and were popping strings left and right.
Our ears are much more accurate in the high range than in the low. For example, a violin player must put his fingers in exactly the right place or we all notice, but if the bass player's fingers are within a half inch of where they're supposed to be it's close enough. Bear this in mind, and always tune your highest string first.
Besides the few specifics already given, human hearing is too vastly complex for us to deal with in any completeness here. Suffice it to say: hearing varies from person to person, with temperature and humidity, and as we get older we tend to lose it (especially in the upper registers).
First, make sure that your guitar is adjusted properly and has good strings. Then there are two true paths:
1) Equal temperament. Buy an electronic tuner, use it and trust it. The guitar won't sound perfect to a very good ear but it will sound fine to most of us, and you can switch keys with impunity. If you don't have an electronic tuner, tune your high E to whatever is available and then find an E on every string and match it to the high E. Don't bother trying to use harmonics! Harmonics are perfect pitches above an equal temperament fingerboard.
2) Approximating Perfect. Start with the electronic tuner and then adjust your tuning using your tuning pegs or by bending individual notes in a chord as you play until the sound pleases your ear. Try tuning to perfect pitches where you are going to spend most of your time in a song. For example: in the key of C tune the C chord in first position to a perfect C chord. The F and G chords will be fine and when you go up the neck to play lead, bend into tune. If a perfectly in-tune note at the bottom of the neck yields a sharp note up the neck tune it low and bend it. Or, play fast and no one will notice.
Steve Mason Luthiers
3809 W. 24th Street
Lawrence, KS 66047-2508


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Tuesday, April 5, 2011
And Thus Spoke Media Blasters...
The following was taken from the official Media Blasters Facebook page on April 5, 2011 at 11:56 AM, PST:
Curiouser and curiouser...
(Please note that the poster of the initial inquiry to Media Blasters, makes an erroneous statement which asserts that MEGALON is slated for release "this year", and attributes that statement to me — and I never posted such a thing! To paraphrase Art Linkletter, "People say the darndest things!")
UPDATE: We've heard back from Media Blasters — now that the channels are open — and I'm asking for an official statement from them to clear the air once and for all. Stay tuned...
5 comments:
- rogue evolent said...
You are such a "consumate pro" August... I really respect the way you've been handling this one!
- Jay - April 5, 2011 at 3:09 PM
- August Ragone said...
August - April 5, 2011 at 7:51 PM
- blockhouse said...
August, last night I was was told an alternate take on your Megalon story. This one states that Elvin Feltner, owner of the then Krypton International Films, Inc. bought up the rights to Megalon years ago. There may be some truth to that as Krypton International is listed as underlying rights owner on The Mystery Science Theater 3000 collection Volume 10 put out by Rhino Home Video in 2006. Now a company called Film Chest says that in 2008, films from the film library of of Elvin Feltner, described by them as a legendary West Palm Beach Florida film collector were saved "days from being in the dumpster"according to Philip Hopkins, co-owner of Film Chest in Rockport, Massachusetts, that Film Chest now have the rights to the the Felter/Krypton archive and have started to restore and release some films. Nowhere on their websight, www.filmchest.com can I find a reference to Godzilla Vs Megalon. One other thought was that Film Chest was going to collaborate with Media Blasters to release the film, which brings the story full circle.
- April 9, 2011 at 7:38 AM
- August Ragone said...
Interesting. Very interesting... - April 16, 2011 at 4:23 PM
- brownsean said...
August, you probably already know this but Blu-Ray.com posted this news blurb a few days back..."Media Blasters released news today of their upcoming Destroy All Monsters and Godzilla vs. Megalon Blu-rays." No release date yet for Megalon but D A M presumably October...
- June 1, 2011 at 8:39 AM


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Guts go a long way at Sawgrass, meaning Graeme McDowell should go a long way at Sawgrass. The Ulsterman displayed his customary fortitude yesterday to remain in contention at the halfway point of The Players Championship.
A 69 hauled McDowell to eight-under and two behind the leader David Toms. It was a satisfactory manner in which to follow up his opening 67, although what made it truly commendable was the fashion in which he bounced back from his back-nine mishaps. Three times he fell foul of the scorecard, and three times he responded with a birdie. The most notable came on the feared 18th when he guided in his approach to three feet.
That helped rid his memory bank of an unseemly double-bogey six on the 12th, the shortest par four on the course. Accuracy is everything off that tee, as McDowell discovered. But then, on the par-three 13th he struck his iron to 18 inches and so the narrative was set. That was one of seven birdies for the world No 5, who believes his game has finally clicked after a session with his coach, Pete Cowen. Three missed cuts in four events have inevitably played heavily on his mind.
"Crazy things go through your head, like 'Will I ever been in contention again, am I done, am I finished?'" he confessed. "Sports psychologists say don't wait for the golf ball to make you happy; try and go out there and be happy. But the way my golf ball has been flying, it's been pretty tough to be happy. So I realise my technique has been the biggest problem. I really have been swinging it badly. It feels good to go out there and make some swings, see the shots, execute them. It gives me confidence."
If McDowell can reduce the error count at the weekend then he could well lift his third American title in a year, having began his dream run on Stateside with victory in the US Open. This is such a welcome return to form with his major defence just five weeks away.
However, "the fifth major" – as some Americans quaintly call The Players – is still a big ask. In a tie for third alongside McDowell is his Ryder Cup partner Luke Donald. Lee Westwood may be at Wembley today watching the FA Cup final, and Donald is in Florida. But believe it, the latter will have the former in his sights. A win would give Donald the world No 1 berth currently held by Westwood and on the evidence of yesterday's 67 that possibility is a live one. Rather incredibly, he became the first player in seven years to avoid taking a bogey in the first two rounds at The Players.
And then there is Tiger Woods, also absent but also still very much a talking point. After his withdrawal following just nine holes in the opening round, doubts continue to intensify over his participation in the US Open at Congressional and July's Open Championship at Sandwich. Woods, himself, was unable to put any sort of timeframe on his return from his latest injury to his left knee, causing yet more consternation.
"I think his whole future is up in the air," said Butch Harmon, who was his coach for 10 years. "Does this mean he needs more surgery? If he does, who knows what will happen."
Harmon's concern is understandable. Woods has undergone four operations on his left knee, the last of which, three years ago, was radical reconstruction which was supposed to solve the weakness once and for all. It has blatantly failed on that score. He was sidelined for four and a half weeks after injuring his knee and Achilles at The Masters, but the break was clearly not long enough.
- More about:
- Golf
- Lee Westwood
- The Masters
- US Open- Golf


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1 Routes from Anantapur to Bangalore by Road
On this Route - the Distance from Anantapur to Bangalore is 218 km.And Drive from Anantapur to Bangalore takes about 4.36 hours by car.
Go Straight on NH 7 cross Nyamaddala, Penulonda, Budili
Continue on NH 7
Continue on NH 7 cross Devanahalli
at Bangalore
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Popular tourist places from Bangalore
Shravanabelagola (142 kms)Mysore (145 kms)Srirangapatna (125 kms)Yelagiri (141 kms)Kanakapura (56 kms)


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Redirected from Brain-in-a-jar
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Brain-in-a-Jar | |
Type | Bookshelf |
Physics | No |
Tool | |
Stackable | Yes |
Mod Included | Thaumcraft |
What else can I say? It's a brain in a jar. It does occasionally whisper something useful - maybe you should use it instead of bookcases for enchanting or research...
Usage Edit
It functions identically to a Bookcase for Enchantment Tables, Thaumic Enchanters, Quaesitums and Occultic Enchanters. Additionally, it improves the speed of Occultic Enchanters and reduces the failure rate and loss rate of a Quaesitum.
Recipe Edit
On some older versions of Thaumcraft, this recipe may work:
Notes Edit
- The Dark Infuser discovery is a prerequisite to getting the Brain-in-a-Jar theory.
- This discovery can only be made researching Zombie Brains. It is a prerequisite for the Occultic Enchanter.
- It is classified as Forbidden Knowledge.
- When the player approaches, the brain will turn towards them.