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The hypothalamus-pituitary-adrenal (HPA) axis is activated during an immune challenge to liberate energy and modulate immune responses via feedback and regulatory mechanisms. Inflammatory cytokines and prostaglandins are known contributors to HPA activation; however, most previous studies only looked at specific time points following LPS administration. Since whole bacteria have different immune stimulatory properties compared with LPS, the aim of the present studies was to determine whether different immune products contribute to HPA activation at different times following live Escherichia coli challenge. Sprague-Dawley rats were injected intraperitoneally with E. coli (2.5 × 107 CFU) and a time course of circulating corticosterone, ACTH, inflammatory cytokines, and PGE2 was developed. Plasma corticosterone peaked 0.5 h after E. coli and steadily returned to baseline by 4 h. Plasma PGE2 correlated with the early rise in plasma corticosterone, whereas inflammatory cytokines were not detected until 2 h. Pretreatment with indomethacin, a nonselective cyclooxygenase inhibitor, completely blocked the early rise in plasma corticosterone, but not at 2 h, whereas pretreatment with IL-6 antibodies had no effect on the early rise in corticosterone but attenuated corticosterone at 2 h. Interestingly, indomethacin pretreatment did not completely block the early rise in corticosterone following a higher concentration of E. coli (2.5 × 108 CFU). Further studies revealed that only animals receiving indomethacin prior to E. coli displayed elevated plasma and liver cytokines at early time points (0.5 and 1 h), suggesting prostaglandins suppress early inflammatory cytokine production. Overall, these data indicate prostaglandins largely mediate the early rise in plasma corticosterone, while inflammatory cytokines contribute to maintaining levels of corticosterone at later time points.
- corticosterone
- prostaglandin
- indomethacin
activation of the hypothalamus-pituitary-adrenal (HPA) axis is one of the critical brain-mediated sickness responses that enhance survival of an organism during an immune challenge. The resulting elevation in circulating glucocorticoids, mainly cortisol in humans and corticosterone in mice and rats, liberate energy necessary to mount a fever and have numerous immunomodulatory effects that reduce the risk of septic shock and increase the chances of survival during infection (52, 53). For example, glucocorticoids suppress proinflammatory cytokines (2, 17, 34) and prostaglandin production (27, 38), stimulate anti-inflammatory cytokine production (16, 20), upregulate Fc receptors and major histocompatibility complex class II molecules on phagocytes (24), increase cell adhesion molecules on endothelial cells (57), enhance acute phase changes in the liver (3), and mobilize immune cells to sites of infection (10, 40). However, it remains unclear how and when various immune factors contribute to the initiation and maintenance of HPA activation during an immune challenge.
Numerous studies have investigated the role inflammatory cytokines, TNF-α, IL-1, and IL-6 play in HPA activation (11, 55) since these are the first cytokines released following immune activation (typically within 60–90 min). Studies generally find that inflammatory cytokines are sufficient, but not necessary, to activate the HPA axis during immune challenge. Administration of any one of the inflammatory cytokines potently stimulates the HPA axis and results in elevation of circulating corticosterone (11, 55). However, blockade of any individual inflammatory cytokine via administration of a receptor antagonist (12, 13, 50), neutralizing antibodies (46, 60) or use of selective cytokine or receptor knockout animals (31, 37, 58) fails to block HPA activation following LPS challenge. IL-6 appears to play the largest role since administration of IL-6 neutralizing antibodies (60), and IL-6 deficient mice (58) show some attenuation of corticosterone levels following LPS administration; however, a robust HPA response is still observed in these animals. The overall conclusion has been that all the inflammatory cytokines contribute to HPA activation and blockade of any individual cytokine fails to prevent HPA stimulation due to duplicity in their actions.
More recently, focus has turned to the role prostaglandins play in activation of the HPA axis during an immune challenge. In contrast to the soluble protein cytokines, prostaglandins are lipid mediators that can be rapidly (within minutes) released following immune stimulation via the conversion of arachidonic acid to PGH2 by constitutively expressed cyclooxygenase-1 (COX-1) and then converted to PGE2 by PGE synthase; furthermore, later production of PGE2 can be synthesized by the inducible COX-2 (39). Administration of PGE2 is sufficient to stimulate the HPA axis (43, 61, 62), but in contrast to cytokines, a nonselective COX inhibitor completely blocks the early (0.5–1 h) corticosterone response after LPS challenge (14, 23). Similar to inflammatory cytokines, prostaglandins are released by innate immune cells in local tissue and have been shown to signal the brain via activation of peripheral nerves such as vagal afferents in the liver (36) or released directly by endothelial cells composing brain microvasculature (23, 49, 51). The rapid production of prostaglandins compared with soluble proteins following an immune challenge led to the hypothesis that PGE2 mediates the initial activation of the HPA axis, while inflammatory cytokines help maintain HPA activation at later time points.
We present a series of studies that examined the contribution of both lipid (i.e., prostaglandin) and protein (i.e., IL-6) immune products across time in mediating activation of the HPA axis following intraperitoneal injection of live Escherichia coli. Previous studies used purified LPS derived from the cell membrane of gram-negative bacteria to stimulate immune responses, but LPS is only one component in gram-negative bacteria cell membrane that selectively targets toll-like receptor-4, while whole bacteria possesses multiple pathogen-associated molecular patterns capable of activating several toll-like receptors and is more potent at activating other immune responses, such as complement proteins (5). Here, studies examined corticosterone levels at multiple time points (30, 60, 120, 240 min) following two concentrations of E. coli, and used indomethacin (a nonselective COX inhibitor) and IL-6 neutralizing antibodies to examine the contribution of prostaglandin and IL-6 in activation of the HPA.
Adult male Sprague-Dawley rats (175–199 g; Harlan) were used for all studies in this investigation. Animals were single housed in Plexiglas cages with food and water available ad libidum and a 12;12-h light-dark cycle beginning at 07:00. Animals were given 1 wk to adjust to their new environment before any study was performed. Animals were handled once a day in the morning for 4 days before any study was performed to help minimize their stress response during the studies. All studies began in the morning (∼09:00–10:00 h) to minimize the effects of the natural rise of corticosterone in the early evening. Care and use of the animals was in accordance with protocols approved by the Kent State University Institutional Care and Use Committee. All studies were designed to minimize pain and the number of animals used.
Drug Concentration and Administration
Indomethacin (Sigma-Aldrich), a nonselective COX inhibitor, was injected at a concentration of 5 mg/kg in a volume of 100 μl of sterile 33% dimethyl sulfoxide. Rat IL-6 antibody (cat. no. AF506; R&D Systems, Minneapolis, MN) was injected at a concentration of 4.2 μg/kg in a volume of 250 μl of sterile saline. All drugs were injected intraperitoneally.
Bacterial Growth
E. coli (0111:B4; ATCC 15746; American Type Culture Collection) was rehydrated and grown overnight in 40 ml of brain-heart infusion broth (DIFCO Laboratories) at 37°C at 5% CO2. Bacterial cultures were then aliquoted into 1 ml brain-heart infusion broth with 10% glycerol and frozen at −20°C. All studies used bacteria from these stock cultures. One day prior to experimentation, stock cultures were thawed and cultured overnight in 40 ml of brain-heart infusion broth at 37°C and 5% CO2. Bacteria were quantified by extrapolating from previously determined growth curves. Cultures were centrifuged for 20 min at 3,000 rpm, and supernatants were discarded. Bacteria were resuspended in sterile 0.9% saline at a concentration of 1 × 108 CFU/ml for studies 1, 2, and 3, and at a concentration of 1 × 109 CFU/ml for studies 4 and 5. Rats were injected intraperitoneally with 250 μl of E. coli (2.5 × 107 CFU for studies 1, 2, and 3 and 2.5 × 108 CFU for studies 4 and 5).
Blood Collection
For studies 1 and 5, rats were euthanized by decapitation and truck blood was collected in 10 ml EDTA-coated vacutainer tubes (BD PharMingen) and stored on ice until the completion of the study. Blood was centrifuged for 10 min at 4,000 rpm at 4°C, and plasma was collected and stored at −80°C until the time of assay. For studies 2, 3, and 4 tail vein blood was collected at designated time points. Rats were gently wrapped in a towel and lightly secured with a Velcro strap prior to tail blood collection. A lateral tail vein was nicked at the posterior end of the tail using a scalpel blade, and the tail was stroked from anterior to posterior to facilitate the movement of blood. Approximately 200–300 μl of blood was collected in an Eppendorf snap-cap tube within ∼60–90 s of removing the animal from the cage. Blood was centrifuged for 10 min at 10,000 rpm, and plasma was collected and stored at −80°C until time of assay.
Tissue Collection
For studies 1 and 5, a small specimen of liver was quickly harvested from the rats and frozen in liquid nitrogen, and then stored at −80°C until time of use. Each liver specimen was sonicated in Iscove's Dulbecco Medium with 2% aprotinin for ∼10 s. Specimens were centrifuged at 10,000 rpm for 10 min at 4°C and the supernatant was collected for measurement of cytokines (IL-1β, IL-6, and TNF-α) by ELISA. Cytokine measurements were corrected for protein by Bradford assay.
Measurement of Peripheral Mediators
Cytokines were measured using available ELISAs for IL-1, IL-6, and TNF-α (R&D Systems). Plasma ACTH also was measured by ELISA (ALPCO). PGE2 and corticosterone were measured by enzyme immunoassay (Cayman Chemical and Assay Designs, respectively). All ELISAs and enzyme immunoassays were performed according to manufacturer's recommendations. PGE2 samples were diluted 1:20 in the manufacturer's enzyme immunoassay buffer, and corticosterone samples were diluted 1:50 in dH2O and heated for 1 h in 70°C water bath.
Experimental Design
Study 1.
Animals (n = 6–8/group) were injected intraperitoneally with either saline or E. coli (2.5 × 107 CFU) and euthanized by decapitation 0.5, 1, 2, 4, or 24 h later. One or two saline-control animals were euthanized at each time and presented as a 0-h time point. Trunk blood was collected for measurement of cytokines, ACTH, corticosterone, and PGE2. A small specimen of liver was quickly harvested, and cytokines was measured by ELISA.
Study 2.
A baseline sample of tail vein blood was collected prior to drug and E. coli administration. Animals (n = 6/group) were injected intraperitoneally with either vehicle or indomethacin (5 mg/kg) 30 min prior to intraperitoneal E. coli administration (2.5 × 107 CFU). Tail vein blood was then collected at 1, 2, and 4 h after E. coli challenge for measurement of corticosterone. PGE2 was also measured in blood samples to verify blockade of PGE2 formation by indomethacin.
Study 3.
A baseline sample of tail vein blood was collected prior to drug and E. coli administration. Animals (6–9/group) were injected intraperitoneally with either saline or IL-6 antibody (4.2 μg/kg) 30 min prior to intraperitoneal E. coli administration (2.5 × 107 CFU). Tail vein blood was collected at 1, 2, and 4 h after E. coli challenge for measurement of corticosterone.
Study 4.
A baseline sample of tail vein blood was collected prior to drug and E. coli administration. Animals (6–9/group) were injected with either DMSO or indomethacin 30 min prior to intraperitoneal E. coli challenge (2.5 × 108 CFU). Tail vein blood was collected at 1, 2, and 4 h after E. coli challenge for measurement of corticosterone.
Study 5.
Animals (6–9/group) were injected with either DMSO or indomethacin 30 min prior to intraperitoneal saline or E. coli (2.5 × 108 CFU) administration. Animals were killed by decapitation at 0.5 and 1 h, and trunk blood was collected for measurement of cytokines, corticosterone, and PGE2 by ELISA or enzyme immunoassay. A small specimen of liver was quickly harvested for measurement of tissue cytokines.
Statistical Analysis
Data from study 1, which examined the effects of E. coli on various mediators of HPA axis activation, were analyzed using a 1 × 6 ANOVA across time (0, 0.5, 1, 2, 4, and 24 h). In studies 2, 3, and 4, tail blood was taken from each animal to examine the effects of drug administration on corticosterone release across time. A repeated-measures ANOVA was used to examine the effects of each treatment across time (0, 1, 2, and 4 h), and significant interactions were further analyzed using one-way ANOVAs at each time point. Necessary post hoc analyses were done using Fisher's least significant difference test. P < 0.05 was used in all cases as a measure of significance to reject the null hypothesis.
Study 1: Time Course of Mediators Involved in HPA Activation
To establish a time course of HPA activation following E. coli administration and to determine the factors that might be involved in HPA stimulation at each time point, animals were injected intraperitoneally with 2.5 × 107 CFU of E. coli or saline and killed 0.5, 1, 2, 4, and 24 h later. After E. coli challenge, plasma corticosterone levels peaked at 0.5 h and decreased in a time-dependent manner, thereafter returning to baseline levels by 4 h [F(5,44) = 14.720, P < 0.001]. Post hoc analysis revealed that corticosterone was significantly elevated at 0.5 h (P < 0.001), 1 h (P < 0.001), and 2 h (P = 0.020) following E. coli challenge compared with saline-injected controls (0 h) (Fig. 1A). Plasma ACTH also increased across time [F(5,42) = 7.655, P < 0.001] following E. coli challenge. ACTH began to increase at 0.5 h (P = 0.072) although levels did not reach significance until 1 h (P < 0.001) and returned to baseline levels by 2 h (Fig. 1B).
Plasma PGE2 showed a similar pattern following E. coli challenge as plasma corticosterone with peak levels at 0.5 h and levels slowly decreasing across time [F(5,42) = 17.318, P < 0.001]. Post hoc analysis revealed that plasma PGE2 is rapidly elevated by 0.5 h (P < 0.001), continues to be significantly elevated at 1 h (P = 0.004), and returns to baseline levels by 2 h. Interestingly, PGE2 significantly decreased at 4 h (P = 0.015) compared with baseline levels before returning to basal levels by 24 h (Fig. 1C). Not surprisingly, the inflammatory cytokines took longer to elevate in plasma following immune challenge. Plasma IL-6 levels increased across time [F(5,45) = 53.588, P < 0.001] with levels significantly elevated at 2 h (P < 0.001) (Fig. 1D), but no longer significantly elevated at 4 h (P = 0.072). Plasma TNF-α also increased across time [F(5,43) = 11.723, P < 0.001] with TNF-α levels elevated at 1 h (P = 0.193) and significantly elevated by 2 h (P < 0.001), and returned to baseline levels by 4 h (Fig. 1E). Plasma IL-1β was measured, but there were no significant interactions across time [F(5,42) = 1.768, P = 0.141] with only low levels of IL-1β being detected (figure not shown).
Liver IL-1β showed a significant interaction across time [F(5,45) = 8.840, P < 0.001] following E. coli challenge. Post hoc analysis revealed that liver IL-1β levels did not rise until 2 h after E. coli challenge (P < 0.001). Liver IL-1β was still elevated at 4 and 24 h, but the values were not significant (P = 0.075 and P = 0.357, respectively) (Fig. 1F). Liver IL-6 and TNF-α levels were not elevated at any time point [F(5,47) = 1.189, P = 0.331] and [F(5,47) = 0.318, P = 0.899], respectively (figures not shown).
Study 2: Effects of Prostaglandins on HPA Axis Activation
Based on study 1, the time course of plasma PGE2 best matched that of plasma corticosterone as both were shown to rapidly elevate following E. coli challenge, with peak values reached within 30 min. To examine the role PGE2 may play on HPA activation and ultimately corticosterone secretion, animals were injected with either DMSO or indomethacin prior to E. coli administration, and plasma corticosterone was measured across time. A repeated-measures ANOVA revealed a significant effect of treatment across time [F(2,17) = 16.354, P < 0.001]. Administration of E. coli resulted in a rapid increase in plasma corticosterone with the greatest increase at 1 h and then levels steadily decreasing across time. Indomethacin alone had no significant effects on plasma corticosterone at any time point, but completely blocked the rise in corticosterone 1 h after E. coli challenge, but not 2 h following E. coli challenge (Fig. 2A). To verify that the effects of indomethacin did not wear off by 2 h, plasma PGE2 levels were measured at 1 and 2 h after E. coli challenge. Indomethacin completely blocked the rise in plasma PGE2 at both 1 h [F(2,17) = 11.305, P = 0.001] and 2 h [F = (2,14) = 15.525, P = 0.001] following E. coli challenge (Fig. 2B).
Study 3: Effects of IL-6 on HPA Axis Activation
As demonstrated in study 2, plasma corticosterone was elevated at 2 h following E. coli challenge, despite continual blockade of plasma PGE2. Since plasma IL-6 was significantly elevated at 2 h and IL-6 has previously been shown to contribute to LPS-induced HPA activation, we examined the contribution of IL-6 on E. coli-induced plasma corticosterone via immunoneutralization techniques. Administration of rat IL-6 antibody alone had no significant effects on plasma corticosterone compared with baseline levels across time; however, administration of IL-6 antibody significantly attenuated plasma corticosterone levels at 2 h following peripheral E. coli. A repeated-measures ANOVA revealed a significant effect of drug treatment across time [F(2,17) = 30.023, P < 0.001] with post hoc analyses revealing that administration of IL-6 antibody only attenuated plasma corticosterone levels 2 h after E. coli challenge (P = 0.004) (Fig. 3).
Study 4: Effects of Prostaglandins on HPA Activation After Higher Concentration of E. coli.
To further investigate the role of PGE2 in mediating HPA responses to higher concentrations of bacteria, we injected animals with either DMSO or indomethacin prior to a 10-fold higher concentration of E. coli (2.5 × 108 CFU) and measured plasma corticosterone across time. Administration of E. coli resulted in high levels of plasma corticosterone at 1 and 2 h with levels slightly lower at 4 h. Unlike with the lower concentration of E. coli, indomethacin only attenuated plasma corticosterone at 1 h (P = 0.044) compared with animals injected with E. coli alone as corticosterone levels were significantly elevated compared with baseline levels (Fig. 4). There were no other significant differences between the two treatments at any other time point.
Study 5: Effects of Indomethacin on Cytokines Production Following a Higher Concentration of E. coli.
Because indomethacin only attenuated the early HPA response after a high concentration of E. coli, we further investigated possible mediators that could be responsible for early activation of the HPA axis following a high concentration of E. coli. Animals were either injected with DMSO or indomethacin 30 min prior to intraperitoneal saline or E. coli administration, and animals were killed 0.5 h or 1 h later. Again, E. coli administration significantly elevated plasma corticosterone compared with DMSO-injected controls at both the 0.5 h (P = 0.001) and 1 h (P < 0.001) time points. There was a significant interaction between the treatments at both time points [F(2,15) = 8.351 P = 0.005 and F(2,14) = 22.812, P < 0.001, respectively] with post hoc analysis revealing animals injected with indomethacin prior to E. coli challenge had significantly attenuated plasma corticosterone levels at both 0.5 h (P = 0.024) and 1 h (P = 0.001) compared with E. coli-injected animals (Fig. 5A). E. coli administration also significantly elevated plasma ACTH compared with DMSO-injected controls at both the 0.5 h (P < 0.001) and 1 h (P < 0.001) time points. There was a significant interaction between the treatments at both time points [F(2,16) = 15.799, P < 0.001 and F(2,19) = 14.721, P < 0.001, respectively] with post hoc analysis revealing that indomethacin administration prior to E. coli challenge significantly attenuated plasma ACTH levels at both 0.5 h (P < 0.001) and 1 h (P < 0.001) compared with E. coli-injected animals (Fig. 5B).
Plasma PGE2 also showed a significant interaction between treatments after E. coli challenge at 0.5 h [F(2,17) = 7.926 P = 0.004] and 1 h [F(2,17) = 6.142, P = 0.010]. Post hoc analysis revealed animals injected with E. coli had significant increases in plasma PGE2 compared with control animals at both 0.5 h (P = 0.008) and 1 h (P = 0.011), and indomethacin completely blocked the elevations in PGE2 at both time points (Fig. 5C). Similar to that observed following the low concentration of E. coli presented above in Fig. 1, plasma TNF-α, IL-1β, and IL-6 were not significantly elevated at 0.5 h or 1 h following the high concentration of E. coli. Interestingly, a significant effect of treatment was observed for plasma IL-1β at 1 h [F(2,17) = 4.066, P = 0.035], plasma IL-6 at both 0.5 h [F(2,17) = 4.057, P = 0.039] and 1 h [F(2,17) = 10.007, P = 0.001], and plasma TNF-α at both 0.5 h [F(2,17) = 6.816, P = 0.008] and 1 h [F(2,17) = 8.537, P = 0.004]. In all cases, animals injected with indomethacin prior to E. coli challenge had significantly elevated plasma cytokines compared with control animals and animals injected with E. coli in the absence of indomethacin (Fig. 5, D–F).
Liver IL-β showed a similar pattern with a significant effect of treatment at 0.5 h [F(2,14) = 6.080, P = 0.015] and 1 h [F(2,14) = 14.814, P < 0.001]. Indomethacin treatment prior to E. coli significantly increased liver IL-1β at 0.5 and 1 h compared with controls (P = 0.005 at 0.5 h and P < 0.001 at 1 h) and animals receiving E. coli in the absence of indomethacin (P = 0.047 at 0.5 h and P = 0.011 at 1 h) (Fig. 6A). Liver IL-6 displayed no significant interactions between treatments at either time point (Fig. 6B). No elevations in liver TNF-α were reported at the 0.5-h time point, but there was a significant difference between treatments at 1 h [F(2,17) = 4.743, P = 0.022]. Post hoc analysis again revealed that only animals injected with indomethacin prior to E. coli challenge had elevations in liver TNF-α at 1 h, and these elevations were significant compared with both control animals (P = 0.015) and animals injected with E. coli alone (P = 0.017) (Fig. 6C).
The present data demonstrate a shift across time in the factors that mediate HPA activation following bacterial challenge. HPA responses following peripheral E. coli challenge are extremely rapid, within 30 min, and occur prior to detection of circulating or liver inflammatory cytokines. Data presented here demonstrate the initial activation of the HPA axis is largely mediated by COX-induced prostanoid synthesis. Not only did the early rise in circulating corticosterone correlate with the rise in circulating PGE2, but the administration of indomethacin, a nonselective COX inhibitor, completely blocked the early rise in plasma corticosterone following a low concentration of peripheral E. coli and attenuated the early rise in plasma corticosterone following a high concentration of E. coli. Interestingly, corticosterone levels peaked long before peak circulating ACTH, suggesting prostaglandins may stimulate corticosterone release through an ACTH-independent mechanism. This would support previous work demonstrating PGE2 directly mediate corticosterone release from cultured rat adrenals (41) and cortisol release from human adrenal H295R cells (56).
Even while prostaglandin production was completely inhibited, corticosterone levels increased 2 h following E. coli challenge, indicating other factors stimulate HPA responses at this time. The later rise in corticosterone correlated with the rise of inflammatory cytokines in plasma (IL-6 and TNF-α) and in liver (IL-1β). Administration of rat IL-6 antibodies attenuated the later rise in corticosterone 2 h following E. coli challenge. It should be noted that the magnitude of the corticosterone response in the original time course study was less compared with subsequent studies for unknown reasons. One possibility is that some bacteria were lost during the decanting process for the original study, which would have reduced the number of actual E. coli injected. However, this does not change the interpretation of the individual studies, that following bacterial challenge prostaglandins largely initiate HPA responses, while inflammatory cytokines, at least IL-6, are likely mediators that maintain HPA activation at later time points.
Most studies in the literature have utilized purified LPS, a component in the cell membrane of gram-negative bacteria, to stimulate immune responses and examine HPA activation. Here, we chose to use live E. coli to more closely mimic a true bacterial challenge as whole bacteria have multiple antigenic epitopes that can stimulate a wider range of immune responses (5). In fact, the time course of HPA activation presented here following E. coli is different than that commonly reported following LPS challenge. Administration of E. coli resulted in a very rapid rise in circulating corticosterone that peaked ∼30 min after E. coli challenge and decrease thereafter, while studies that administer LPS report a slower rise in circulating corticosterone that peaks ∼2 h after challenge (regardless of dose and route of administration) (13, 23, 26, 32, 35, 60). While there appears to be a rather dramatic shift in the kinetics of HPA activation between E. coli and LPS, the mechanisms mediating HPA responses appear to be very similar. By use of genetic and pharmacological techniques, recent studies have demonstrated inhibition of COX-1 results in a significant reduction in corticosterone levels in the first hour after LPS challenge but has little effect on corticosterone thereafter (14, 15, 23). Our data demonstrate COX-mediated prostanoid synthesis is also critical for the initial induction of corticosterone following E. coli challenge, particularly following lower concentrations of bacteria. However, because of the kinetic differences between E. coli and LPS, this study reveals prostaglandins may play a larger role in HPA activation during a bacterial infection than suggested by LPS studies. In studies using LPS, COX inhibitors only block a very small fraction of the overall corticosterone response because very little corticosterone is released in the first hour following LPS challenge (14, 23), while following E. coli the bulk of the corticosterone response is observed within the first hour, especially following lower concentrations of bacteria.
At later time points, inflammatory cytokines likely help maintain HPA responses. Many studies have examined the role of inflammatory cytokines and found that genetic or pharmacological blockade of individual inflammatory cytokines have little effect on attenuating HPA responses to LPS (12, 13, 31, 37, 46, 50, 58), likely due to duplicity in inflammatory cytokine signaling. One exception has been that the pretreatment with IL-6 antibodies often attenuates ACTH (46) and corticosterone (60) responses 2–4 h following LPS but not at early time points (18, 60). Here, we replicate this finding following E. coli challenge by demonstrating administration of goat anti-rat polyclonal IL-6 antibodies attenuate corticosterone levels 2 h following bacterial challenge. Together, these findings suggest prostaglandins mediate early HPA activation and IL-6 (along with potentially other inflammatory cytokines) help maintain HPA activation as needed.
Indomethacin failed to completely block the early rise in corticosterone following higher concentrations of E. coli, suggesting additional mediators may be involved in HPA activation as bacterial challenge increases. We examined whether a higher concentration of E. coli would stimulate a more rapid increase in inflammatory cytokines; but this was not the case as there was no significant increase in any circulating or liver inflammatory cytokines at either 30 or 60 min following the higher dose of E. coli. However, indomethacin administration resulted in exaggerated inflammatory cytokine responses to E. coli challenge as demonstrated by significant increases in liver IL-1β at 0.5 and 1 h, plasma IL-6 at 0.5 and 1 h, and increases in plasma TNF-α at 0.5 and 1 h and liver TNF-α at 1 h. This suggests inflammatory cytokines may elevate in an active compensatory fashion to stimulate corticosterone production, thereby, explaining why indomethacin only attenuates corticosterone responses at 0.5 and 1 h following a high concentration of E. coli. However, the cytokine levels at 0.5 h are very low, and it is unclear whether such values would be sufficient to activate the HPA axis. If these low cytokine levels are sufficient to stimulate corticosterone production, then one would have expected a further increase in corticosterone production as cytokine levels increased from 0.5 to 1 h, but this was not observed. One would also have expected a similar enhancement of inflammatory cytokines and attenuation of corticosterone responses in indomethacin-treated animals challenged with the lower concentration of E. coli, yet indomethacin completely blocked the elevation in corticosterone when administered following the lower concentration of E. coli. Our interpretation of these data is that additional factors contribute to HPA activation at higher bacterial concentrations. One possibility is activation of complement. Complement proteins are constitutively produced and continuously circulate in the blood, and following contact with the surface of bacteria complement proteins can be rapidly activated, thus available to rapidly activate HPA responses. Complement products C3a and C5a have been shown to elevate ACTH (21, 22) and activate paraventricular hypothalamic neurons (8) as well as act directly on the adrenal gland (21). Complement products can also trigger prostaglandin synthesis and release by Kupffer cells (45, 47) adding to HPA activation.
The fact that indomethacin pretreatment resulted in exaggerate inflammatory cytokine responses following E. coli challenge indicates that prostaglandin signaling inhibits inflammatory cytokine production. Prostaglandins can directly inhibit inflammatory cytokines, particularly TNF-α, as demonstrated in cultured Kupffer cells (19, 36, 54), peritoneal (1, 63), alveolar (48), and bone-derived macrophage (30, 59). Additionally, the dampened HPA response in indomethacin-treated animals likely reduced glucocorticoid inhibition of inflammatory cytokine production resulting in widespread increases in inflammatory cytokine production. These data are of particular interest because nonsteroidal anti-inflammatory drugs are commonly used for the relief of pain and inflammation, yet chronic use is associated with an increased risk of atherosclerosis and thrombosis (6, 25, 28). It is possible that COX inhibitors create a prothrombotic state by elevating proinflammatory cytokines responses, which upregulate adhesion molecules on endothelial cells promoting leukocyte adhesion and aggregation to vessel walls and increase the production of tissue factor while simultaneously preventing the formation of PGI2 that typically inhibit proinflammatory cytokine production (44, 64) as well as platelet activation (7) and aggregation (4, 9, 29, 33, 42).
Perspectives and Significance
Our studies demonstrate for the first time a complete time course of the possible mediators involved in HPA activation following peripheral E. coli challenge. The data indicate that early PGE2 synthesis is not only responsible for early corticosterone responses following an immune challenge but is also responsible for controlling inflammatory cytokine responses that ultimately themselves fine tune the HPA response. It should not be surprising that extremely rapid mechanisms, even ACTH-independent mechanisms, have evolved to stimulate glucocorticoid production during bacterial infection given their importance in reducing the risk of septic shock and modulating immune responses to increase the chances of survival during infection.
This work was supported by National Institute of Mental Health Grant R21MH-77004 and the Ohio Board of Regents Research Initiative.
No conflicts of interest, financial or otherwise, are declared by the author(s).
Author contributions: Z.R.Z. and J.D.J. conception and design of research; Z.R.Z., V.M.P., R.M.C., and J.D.J. performed experiments; Z.R.Z., V.M.P., and J.D.J. analyzed data; Z.R.Z., V.M.P., and J.D.J. interpreted results of experiments; Z.R.Z. prepared figures; Z.R.Z. drafted manuscript; J.D.J. edited and revised manuscript; J.D.J. approved final version of manuscript.
- Copyright © 2011 the American Physiological Society
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Here are 10 key fantasy observations from Sunday's games, with analysis of the important trends and an eye toward next week and beyond.
Tomlinson absolutely terrific: Entering Week 9, some fantasy leaguers might have started to wonder if LaDainian Tomlinson was beginning to lose some of his luster. In the two games prior to Sunday's game against the Jets, Tomlinson had totaled just 76 rushing yards and had not registered either a rushing or receiving TD. In Week 9, he more than compensated for the pair of disappointing outings. Tomlinson reaffirmed that he is the best player in fantasy football by scoring four times in a game for the first time in his career. It's difficult to fathom that the feat actually might be a first for Tomlinson, who rushed for 107 yards on 25 carries. But it was a clear signal that you should never doubt him, even if he has a quiet outing or two. No player has an excellent game every week.
Porter has clearly re-emerged: Last week, Jerry Porter awoke from a six-game slump to catch six passes for 123 yards and two TDs. But many of his fantasy owners weren't sure if his outstanding performance was a one-time tease against a porous Titans pass defense, or the start of a turnaround. In Week 9, Porter erased many doubts with a solid seven-catch performance that included a four-yard TD reception. Porter finished with just 68 receiving yards, but it is apparent he has stepped forward with Randy Moss playing at less than 100 percent. When Moss' health improves, Porter should truly benefit from playing as a complement to him, as the Raiders are obviously devising more ways to get him the ball.

subset: commoncrawl


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Battle on The Home Front
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Location may be everything, but these days Turks are wishing they were not quite so well endowed. Straddling East and West, the Black and Mediterranean Seas, Islam and Christianity, Turkey is once again caught in the midst of a brewing conflict. The Bush Administration has been pressuring NATO's only Muslim member to provide substantial support for its planned war on Iraq at one point urging the use of five air bases and facilities for up to 90,000 ground troops while ordinary Turks are overwhelmingly against an attack. More than 85% oppose the U.S.-led campaign, and protests are mounting across...


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Trinity Insurance Services Inc. has been purchased by McNeil & Company Inc. Insurance Risk Services out of New York in a move that gives McNeil a strong foothold in the West.
The deal was closed on Nov. 16., and terms were not disclosed.
Trinity, a Southern California-based multiline managing general agent and excess and surplus lines agency, offers an array of services, including bars, taverns and nightclubs, construction, professional liability, and technology.
But the program Trinity President Tom Gassen is most interested in pursuing for McNeil is AdvenSure, a program for fishing and hunting outfitters. The program also covers resorts and lodges, bed and breakfasts, inns, campgrounds, RV parks and even dude ranches.
Gassen, an avid fisherman, said it is a good time to sell, but the 75-year-old plans to stay on with McNeil to help bring on more outfitters in the Pacific Northwest as clients under the AdvenSure program.
"Tom has a home in Oregon and has done business up there for a number of years," said Dan McNeil, president and founder of McNeil. "He's looking at developing a great deal of business up there in the fishing guide industry."
Gassen said there are several requirements for fishing and hunting guides, who are required to undergo continuing education and know first aid. Gassen will maintain an active role in the insurance business, but part of his efforts to build the fishing guide business for McNeil will be to go fishing, he said.
"That's the greatest way to meet a guide," he added.
Gassen started Trinity in 1986, and has been in the insurance business for six decades. Stepping into an easier life has been a long time coming, he said.
"Fifty-seven years, that's enough," Gassen said.
McNeil, which operates in 49 states, has been looking for a while to be able to better represent its accounts in the West.
"Dan's programs are exclusive to McNeil and Co.," Gassen said, noting that Trinity has several exclusive programs. "Between the two firms that gives us nine or 10 exclusive programs with top rated A+ carriers."
McNeil and Gassen have known each other through the industry for roughly 10 years. McNeil started his company in 1990. McNeil offers a variety of lines, including products for fire departments, rescue squads, ambulance companies, car washes and guides and the leisure industry.
"We've been wanting to find a facility that could support our footprint out there in the West coast," McNeil said. "It's a very exciting market. Basically I wanted a presence out there that would allow easier access for the agents to do business with us. We'll be moving underwriting operations (out West) to support all of our programs out there to service both the Pacific and Mountain time zones."
Gassen said he likes being purchased by McNeil, as the firm is hands on with its clients and is "very risk managment and loss control involved in all of their accounts."
McNeil said they plan to beef up the number of employees as well. "We're going to probably be adding," McNeil said.
McNeil dispelled any thoughts of retiring Gassen, and said he plans to keep him on for quite some time, and it's still not clear what if any title Gassen will hold going forward.
"He's 75 years old. I think he's good for another 20 years," McNeil quipped.
While neither man would offer details on the purchase, Gassen noted that the timing of purchase, just before the end of the year, wasn't a factor in the sale, and he called the acquisition deal "satisfactory."
"The big buzz word in mergers and acquisitions has been EBITDA," he said, referring to a common practice in middle market acquisitions in which buyers and sellers establish target prices based on multiples of earnings before interest, taxes, depreciation and amortization. "That's been used a lot since the soft market has been affected things negatively, which may be made mergers and acquisitions more advantageous for the acquirer than seller."
Gassen said he and McNeil did discuss EBITDA, but that "we came up with a merger and acquisition agreement that's satisfactory to both of us, and that's all I'm going to tell you."


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They Left Where to Come Here?
Scott Lucas | Photo: Scott Lucas | August 6, 2013
Blue Jasmine aside, most San Franciscans hail from cities other than NYC.
Woody Allen's Blue Jasmine hinges on Cate Blanchett's physical (and psychological) move from one coast to the other. And while we can grant Allen a certain license to place New York at the center of the universe, no matter how many True Jasmines we have, the actual number of migrants making the JFK to SFO flight is much smaller than you'd expect.
That's according to data we found from the Urban Institute highlighted by The Atlantic in February 2012, and dug up from the IRS for the period of time from 2004 to 2010. In our data visualization, the size of each bubble corresponds to the influx of residents from a given city to San Francisco. We've pulled out the top twenty cities (they're actually the metro areas, which means that SF is counting Oakland and Hayward as well.)
What the data shows is that if Blue Jasmine were true to the trends, Blanchett's character would have only had to make a quick commuter hop from San Jose's Mineta Airport to SFO. Heck, she could have just taken Caltrain. That's because the population influx from the San Jose region dwarfed the flow all other cities. It was more than triple New York's numbers. The next runners up were L.A. and Sacramento. Only then did NYC make an appearance—way down at number four. Stockton sent almost the same amount of people as the Big Apple did—even more surprising given that it has 300,000 inhabitants to New York City's 8 million.
So while New York City may loom large as a cultural beacon, when it comes to San Francisco, if you can make it here, you probably made it from somewhere other than there.
Top Twenty Cities Migrating To SF (2004-2010):
1. San Jose-Sunnyvale-Santa Clara 64,669
2. Los Angeles, CA 38,571
3. Sacramento, CA 23,924
4. New York NY 19,124
5. Stockton, CA 14,954
6. San Diego, CA 14,495
7. Santa Rosa, CA 11,432
8. Boston, MA 8,993
9. Seattle, WA 8,178
10. Chicago, IL 7,906
11. Washington D.C. 6,964
12. Riverside, CA 6,165
13. Modesto, CA 6,037
14. Phoenix, AZ 4,896
15. Portland, OR 4,153
16. Las Vegas, NV 4,060
17. Dallas, TX 3,382
18. Fresno, CA 3,337
19. Philadelphia, PA 3,224
20. Houston, TX 3,208
Have feedback? Email us at firstname.lastname@example.com
Email Scott Lucas at firstname.lastname@example.com
Follow us on Twitter @sanfranmag
Follow Scott Lucas on Twitter @ScottLucas86


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Julian Raines was one of the first Englishmen to volunteer for the international brigade in Spain. The British Secret Service suspect that the flamboyant Raines was recruited for the KGB by the Bolsheviks during his student days at Oxford and send Robert Florry, a struggling young writer, to Spain after Raines with orders to eliminate him. Florry was an old school chum to Raines and had every reason to hate him. The British are not alone on Raines' trail. The ruthless Communist leader in Barcelona believes that the identity of the double agent conceals a powerful and profitable secret.
This is a novel that constantly surprises. (Originally published as The Spanish Gambit.)
©2010 Stephen Hunter (P)2010 Brilliance Audio, Inc.
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Thank you, Mandy. I think it's beautiful too! :-)
Beautiful flower, thanks Wendy.
Mary, I'm not sure who you're talking about....but Ardy lives in Maryland, advia in a European country that I'm not supposed to mention...even though it's an especially beautiful one, and whatnauts lives in Canada. Oh, and I live in Pennsylvania. I went to a university in Florida and I absolutely LOVED Florida. I even hitchhiked back there years later with a friend and had a wonderful time...lots of memorable adventures on the Keys.
One of my favorite things about Florida was the hurricanes...and the rains that would pop up out of the blue, and then disappear just as quickly.
Y'all must all live in the same area. Gail told me some others lived in the Portland area not too far from her. Stay safe from those storms, I'm in Florida, the lightning capitol of the world, literally. And, of course, there are hurricanes. Be prepared if you know one is coming! Stock up!
I'm guessing you got the storm that went through here this afternoon. Hope your lovely flowers all survived the heavy rain and high winds. I think it was headed for you and on up to Pat. Stay safe. Thanks, Wendy.
Oh, the Wendy Flower, the Wendy Flower! How I love them. Doing the Flower Dance! LOL Thanks, Wendy!
Say what??? I lost my comments to both of you. I swear I responded just a short while ago.
whatnauts, I went outside and snapped a picture so I wouldn't have to pick it.
advia, it's not moving at all. LOL!
This seems to move from the center towards outside.
This one must have been picked in the dawn's early light. Very cool.


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Bil'in Filmmaker Barred Entry to Israel Despite Acquittal
Imad Bornat was acquitted over a year ago on charges of assaulting an officer, but Israeli police still regard his case as 'open'.
Although photojournalist Imad Bornat from Bil'in was acquitted over a year ago on charges of assaulting an officer, police still regard his case as "open" and continue to bar him from entering Israel.
For this reason, Bornat will not be able to attend the Tel Aviv screening tomorrow of part of his film in progress, "Five Broken Cameras." The screening is set to take place before a crowd of producers and representatives of television stations from around the world.
According to director Guy Davidi, the film documents five years of struggle against the separation fence and how the repression of demonstrations against it have affected individual lives.
In November 2008, Bornat was severely injured in a car accident, requiring treatments he received in Israel. But in September 2009 his request to enter Israel was denied. Gabi Lasky, an attorney who represents dozens of Bil'in residents, subsequently learned that Bornat's request had been denied for reasons related to "police prevention" - stemming from a criminal record from 2006.
However, in April 2009, Bornat had been acquitted of all the relevant charges. And since September 2009, Lasky's office has contacted various police officials on Bornat's behalf - including the police's legal advisor, the officer in charge of individuals barred from entering Israel and the head of the acquittals section in the verdicts department - requesting that the error be corrected.
Appeals were also made to the military prosecution and a pre-High Court appeal was filed with the State Attorney's Office. All of these appeals were accompanied by the relevant documents, but to no avail.
"According to the existing records of the Israel Police," a police spokesman told Haaretz on Sunday, "the case is still open and with the military prosecution. Once its results are received, the lifting of the prevention order will be considered according to the standard criteria."
The criminal case against Bornat was opened in October 2006, after he had been arrested by Border Police officers following a protest in his village, where he had been taking pictures. Eyewitnesses initially said the officers beat Bornat, but later testified in court that he had been hit in the face by a radio device that fell on him while he was in the car.
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Police are hunting a dog owner after a young girl was bitten while playing at a Doncaster beauty spot.
The incident happened at the play area in Sandall Beat Woods and the six-year-old needed hospital treatment.
The girl was with her auntie and two cousins near the Ranger Station in the woods, at about 2.30pm on Saturday, August 10, at the same time as a man with a brindle-coloured Staffordshire bull terrier, which was not on a lead.
It began barking at the children, and the man reportedly tried to pull the dog away by its collar but it broke free of the man's grip and bit the girl on her left knee.
The girl, from the Intake area, was treated at Doncaster Royal Infirmary for puncture wounds.
Police are keen to trace the owner who was in his late 30s, about 5ft 9ins, of average build, unshaven, with short dark hair slicked back, wearing dark jeans, an off-white short-sleeved shirt, and was said to be of scruffy appearance.
He was riding a plain matt black mountain bike.
Anyone with information about the incident or the dog owner is asked to call South Yorkshire Police on 101 quoting incident number 696 of August 10.


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For LMS, You'd Better Shop Around
It's not uncommon for organizations using learning-management systems to be unhappy with them, according to a new report. Experts say a company's purchasing process may largely determine its future LMS satisfaction.
By Jill Cueni-Cohen
Are you pleased with your company's learning-management system? Nearly 48 percent of respondents to a new survey by Brandon Hall Group on LMS buying and usage trends said they're looking to leave their current platform behind and move to a new provider.
Delray Beach, Fla.-based Brandon Hall Group Chief Strategy Officer Michael Rochelle says there are several reasons why organizations may want to jump ship on their current LMS.
"The challenge with LMS today is the fact that the learning environment has changed tremendously," he says, pointing toward the recent evolutions in technology, including in the cloud, mobile and social-media realms. "On top of these changes, there's the additional challenge of having to appeal to a multi-generational audience."
Dissatisfaction with LMS stems from a variety of issues, ranging from poor reporting features to unreliable customer support, but Rochelle says there are ways HR leaders can ensure their next LMS lasts, but essentially it's all in how they shop.
The first step to shopping for the best LMS for an organization, he says, is to ensure the decision makers have a true understanding of their learning strategy.
"Once you figure out the learning strategy," he says, "it must be clearly aligned with your audience and goals."
Next up is to make a list of requirements needed for a system that will best support that learning strategy, he says.
"Most organizations would like the system to do a thousand different things, but that doesn't make sense," Rochelle says, noting that research has found organizations will largely use their LMS in just three-to-five ways, so HR leaders should identify those situations for their own organizations and then place them in the appropriate order of importance.
Armed with this information, he says, organizations can then rate how close or how far away a provider is to being capable of reproducing an appropriate "use case" through their system, and the closest systems should then be compared in-depth prior to a final choice being made.
"Solution providers and organizations [should] get to know each other well through the process," he says.
IT expert Steve Kerschenbaum, principal consultant and general counsel for the Marick Group, a Phoenix, Md.-based consulting firm, agrees that any LMS should be limited in its scope and usage.
"If you want to see a record that all of your employees took all the mandatory training they needed to have," he says, "an LMS is good at doing that," he says. "But a lot of the modern LMS providers can perform so many other functions."
When first setting out to find an appropriate LMS for their organization, Kerschenbaum says many buyers go in to meetings with vendors "and hear all these possibilities" about how they can assess and appraise employees online.
"But they don't realize that these things are only possible if your organization already understands what it is that they're trying to measure," he says. "Most organizations don't have a formal assessment process, so if your organization buys an LMS that has all that and goes to implement it, this will end up being a lot of work and costing a lot of money."
He says an organization's upper-level management and IT departments should both provide their input on LMS requirements.
"Your IT staff might have certain devices and security they're going to provide," he says. "The more focused you can make your LMS acquisition, the happier you'll be, so state what you need to accomplish in one sentence."
Kerschenbaum adds that it's not always necessary to replace an existing -- yet under-performing -- LMS.
"Before you buy a new one and get rid of the old, go back to your requirements," he says. "See what you used the first time and revisit those and see if they match. If they match, you keep the LMS. But if they're different, you may need to start shopping."
There are also external resources available to help guide buyers through the LMS maze.
Capterra is an online software-reviews service designed to help organizations shop for learning-management systems at no cost, says J.P. Medved, content editor for the Arlington, Va.-based firm.
He says the ideal number of LMS solutions to demo is three to five and recommends prospective buyers keep a demo scorecard that maps out their top-ten LMS requirements. (Capterra has a filterable software directory with 276 products.)
"This will allow you to compare apples to apples," says Medved.
And pricing should not be the driving force behind your final decision, he says.
"A lot of these systems are priced based on the number of learners," he says. "There's no one-time, upfront fee. This allows [the buyer] to scale things so that you don't have to worry too much if you're just starting out. Price is important, but it's kind of secondary to your requirements. After you fulfill those, then you can focus on pricing."
Medved says he considers the demo to be the most important test when winnowing down to your top two choices.
"The biggest differentiator would be your appraisal of the service of the company," he says. "They'll both be good systems, and the product software will be fine, but look into customer service and see how responsive they are."
Ultimately, he advises buyers to dig even further than simply having a conversation with an LMS sales rep.
"Send questions through their help line and their tech support email address," he says. Whether they respond -- and how quickly -- "can really indicate their typical level of service and help differentiate them further."


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"Gorgeous . . . a murder mystery that's also a brilliant meditation on love and memory and loss."—Maile Meloy Elanor Dymott's gorgeous debut tells the story of Alex, a solitary lawyer who has finally found love in the form of his beautiful wife, Rachel. When Rachel is brutally murdered one midsummer night on the grounds of their alma mater, Worcester College, Oxford, Alex's life as he knows it vanishes.
He returns to Oxford that winter, and, through the… (more)
Every Contact Leaves a Trace is an intelligent literary mystery, featuring the kind of tormented narrator that Robert Browning himself might have relished.
Publisher: W. W. Norton & Company (April 29, 2013)
Format: EPUB
Page count: 384 pages
File size: 1.6 MB
Protection: DRM
Language: English


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Written on: 06/02/2013 by alvinshock (1 review written)
I have a Guild D25 that I traded for a parrot back in 1980. I am primarily a bass player, and the thing has sat around in the case for years. It sounds great, but is an animal to play, I think it needs some work to bring it back into shape. I have been playing it a lot more as of late and wanted to purchase something like a Martin. I have not been able to pull the trigger on a new guitar and I think that I have not found one that has the same resonance. After reading all of the comments here, I think it would behoove me to have some repair work done instead. I am sitting on something quite good here based on your input. Thank you much,


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The disease is given antibiotics like erythromycin, azithromycin and clarithromycin. Antibiotics are appropriate for cellulitis condition that has not affected the blood stream with the patient and therefore are not yet widespread. Tazarotene comes in a gel or cream and is employed to keep pores to clean. It may be the drug-of-choice inside treatment of serious methicillin-resistant Staph. Other non-prescription supplements and medicines that may not able to be studied include vitamins, vitamins and minerals, herbal supplements, laxatives, antacids, magnesium, vitamin D, calcium, and phosphate.
Staph aureus is the most typical bacteria, causing more than 80% of cases. Moreover, apart from being convenient, they may be safe. It could be passed between species, including hamsters, rabbits, ferrets, birds, livestock, etc. Taking it with food will assist you to prevent an upset stomach. Consult your dermatologist for your right antibiotic and dosage.
If you are considering organic beef, find the USDA Organic label. For more detailed information, please talk to your local physician. Because Gonorrhea and Chlamydia are really much alike it's sometimes very difficult to determine exactly which disease you have contracted and further intricate testing will have to be carried out to determine this fact, also sometimes it can be possible for the person to contract both of these viruses at any given time. It is usually characterized by the inflammation of parenchyma of lungs and alveolar filling with fluid. The next time you hear your companion complaining how they can't last as long without ejaculating or when your partner says their not horny or they don't really want to have sex, you've got a reason to check out the doctor.
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Product Description
Designer bathroom Diadema where the modules meet and fit together. Furniture with sinuous lines to the mirror and the sink and rigorous lines for the bathroom cabinet and wall units. full of games and voids, muted colors and exquisite materials are the result of creativity and research for an innovative style and timeless.
Suspended bathroom cabinet measures: L. 143,8 P. 53,7 H. 210 cm
Composition modern bathroom furniture:
- Base 2 drawers W. 90 D. 51,4 H. 48 cm
- Base with drawer L. 50 P. 24 H. 51.4 cm
- Based on day L. 50 P. 24 H. 51.4 cm
- Washbasin cristalplant L. 63 P. 43 H. 12 cm
- Wall unit L. 30 P. 16 H. 30 cm
- nr 2 wall units L. 50 P. 16 H. 25 cm
- Mirror with LED Ø 90 cm
Mixer and siphon are not included.
Product Quality
Polished Cristalplant. Tub without overflow hole.
GRASS made of metal runners to return amortized with metal and quick-release latch system cover.
opening with outlet at 45 ° throat on the front.
- laminate: 18 mm panels made of wood particles with a polyurethane bonding edges and internal structure in aluminum gray.
- lacquered: NOVOPAN panels resistant to moisture and internal structure in gray.
- Ash: NOVOPAN repellent E1 class panels, with sides exposed coated melamine with double-layer barrier and interior in gray melamine.
- laminate: 18 mm panels made of wood particles with edges to polyurethane bonding.
- lacquered: MDF panels.
- ash: water repellent NOVOPAN panels E1 class, with sides exposed coated melamine with double-layer barrier.
This company was born in the province of Pordenone, in the 80's, has grown exponentially, such that in 1991 became one of the national leaders of the 'bathroom furniture industry.
Later, with the expansion of the distribution to foreign countries, this manufacturer becomes an important player on the international market for the bathroom player. Another recent expansion and the consequent growth in production has allowed the company to increase the offer, ensuring the ideal solution for every kind of bathroom furniture.
The modularity of the components allows extreme flexibility in design, allowing for a "tailoring" creating your own bathroom. With a range of over 300 finishes the series, the company offers a degree of customization unmatched, innovative materials and increasingly efficient contribute to the realization of the ideal product for every need of furniture and personal taste.


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Might gave the opportunity for half a day at Dalby next week.
Looking at the website the red looks an ideal length at just over 20 miles but seems to be split into 3 sections. I can't see a trail map so was wondering if they all flowed on from one another to form a loop or do they start from different places?
Also what are conditions like if anyone's been recently?


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WILTZ: Have you ever heard of Pheasant City
It's part of the wisdom that comes with age. When I'm going to spend some time outdoors, I pay attention to the weather forecast, and I put considerable thought into appropriate dress. Our vehicle is also equipped for a worst case scenario.
Our South Dakota weather can kill. Not long ago, a Redfield man died from exposure after abandoning his car. He underestimated the severity of the conditions. Don't think for a second that we can't have blizzards like they had in the 1880s. We still have those storms, it's just that our warning systems are much improved.
Like most South Dakotans, Betsy and I can remember a particular storm. We were married in June 1965. I taught at Willow Lake, and we lived in a single-story farm house a mile south of town. It was January and the Pirates had a big basketball game in Huron. We considered going, but Betsy was six months pregnant, so we decided to stay home. A winter storm was the farthest thing from our minds.
When I got up for school the next morning, we couldn't see out any of the windows. We tried the front and back doors, but they were snowed shut. I called school to see if there was school, but there was no answer. We eventually learned that at game's end, skies in Huron were clear. By the time the Willow Lake fans reached the Pheasant City gas station, 18 miles north of Huron at the intersection of Highways 37 and 28, further travel was impossible.
Eighty-three people spent three days at Pheasant City. The pop, chips and candy bars were gone in short order, the toilet ran over as the septic tank and drain field flooded and most of the people got sick. Still, had it not been for Pheasant City, good friends and students might have perished.
On the lighter side, February 1971 marked South Dakota's first Class B state wrestling tournament, which was held in Mitchell. It began to snow Thursday night, and as coach of the Parkston wrestlers, I worried about making it to Mitchell the next morning. Before going to bed that night, I had a fleet of four-wheel drives and snowmobiles ready to go. If you think my imagination is out of control, check with my good friend Jerry Opbroek. Jerry was coaching the Burke wrestlers at the time, and they used snow mobiles to get the Bulldogs to the Corn Palace.
South Dakota winter storms differ from those of Iowa or Minnesota. Our wind and theirs are completely different things. Wind makes cold weather brutal. Murderous would be a better word. We are also far more remote. Farms and ranches are farther apart. A seemingly harmless ice fishing junket to a dam north of Burke or a lake west of Colome might just as well be in the Arctic when the snow starts blowing.
Don't go alone! Take along some warmer clothes and blankets. Fill the gas tank, and carry a spare gas can. Pack some candy bars. Along these lines, I was really frustrated the week of Monday, Feb. 25. Betsy made me promise her that when I went ice fishing, I wouldn't go out on the ice if no one else was there.
On Thursday, Feb. 28, I headed to Pickstown. I was after big northern pike, and I had brought some big, live shiners back to South Dakota from Wisconsin. Most of the ice was out of the bay, and what was left looked rotten. I kept my promise to Betsy as no one was out there. I had the same experience Saturday afternoon at Dante Lake. In spite of the balmy 45 degree weather, no one was out there. I felt cheated, but I was alive and well. Betsy was right in making me promise.
* * * * * * * * * *
Over the years, I've pondered whether or not the Sierra Club was for or against hunting. Sierra Club representatives have personally told me that they were indeed "pro" hunting, but at times I've had my doubts. Still, their founder, John Muir, once wrote, "Fine scenery may not stir a fiber of mind or body, but how quick and how true is the excitement of the pursuit of game!" The guy was right on!
The Sierra Club recently revealed its current true colors. In or about August of 2012, after years of legal battles and injunctions, the Western gray wolf was finally removed from the federal list of endangered species. In the heavy wolf population states of Idaho, Montana and Wyoming, wolf hunting seasons were established. Immediately, new lawsuits were in place to re-list the wolf as endangered. These suits were initiated by The Humane Society, Earthjustice and other groups in spite of the fact that the wolves had decimated many elk herds.
I recently learned ("Backcountry," Sports Afield, March/April 2013) that the Sierra Club is in partnership with the Earthjustice law suit. There is nothing more to say. We now know where today's Sierra Club stands.
* * * * * * ** * * *
Today, I hope you will tolerate a little history in today's column that's far more interesting and meaningful than a few storms I pulled from my memory.
In January, some letters to the editor of The Daily Republic dealt with whether or not President Franklin Delano Roosevelt knew of Japan's plan to attack Pearl Harbor prior to the attack. Though there have long been some strong suspicions, there seemed to be no conclusive proof that he knew until The Freedom of Information Act enabled or forced the Navy to release the incriminating documents. A letter in The Daily Republic suggested reading "Day of Deceit" by Robert B. Stinnet. The glossary of Stinnet's book contains copies of the revealing documents, and I have since read the book.
Roosevelt not only knew of the attack, he, through the implementation of a plan orchestrated by Lt. Commander Arthur H. McCollum, provoked the attack. The McCollum plan included: "Giving all possible aid to the Chinese government of Chiang Kai-shek, Sending a division of heavy cruisers and two divisions of submarines into Japanese waters, Keeping the main strength of the U.S. fleet in the Hawaiian Islands as an inviting target, Insisting that the Dutch refuse to grant Japanese requests for goods including oil, and embargoing all trade with Japan in collaboration with a similar embargo by the British."
Other than American provocation, U.S. cryptographers had broken all of Japan's codes. Our highest-ranking leadership, with the exception Admiral Husband E. Kimmel, commander-in-chief of our Pacific fleet, knew of Japanese plans. Kimmel had been purposely kept in the dark. If Kimmel had been allowed to defend Pearl Harbor as he saw fit, there would have been no Japanese attack.
Before we condemn FDR, we must look at his reasons for wanting World War II. Stinnet actually sympathizes with Roosevelt up to a point. We'll look at the rest of the story next week, along with some thoughts on the meat we eat.


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Wild Horse Customs DAMASCUS NIGHTPROWLER Hunter Knife
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Wild Horse Customs DAMASCUS NIGHTPROWLER Hunter KnifeWild Horse Customs by J. Kidd "Let you passion for knives run wild" FEATURES : WOOD HANDLES GOLD DUST CAT W/RUBY EYES INLAY INDIVIDUALLY SIGNED & NUMBERED PROTOTYPE Wild Horse Customs DAMASCUS NIGHT PROWLER Hunter Knife. Genuine Rosewood handles feature a Gold Dust Cat inlay with ruby eyes. 8 7/8" overall length, 4 5/8" blade. DAMASCUS STEEL. Genuine leather belt sheath. Comes with Certificate of authencity. Prototype. Comes new with Sheath. This is a beautiful Knife!!!!! We accept all forms of payment through PAYPAL, money orders/cashiers check. PLEASE, NO PERSONAL CHECKS. Shipping inside the US is $6.95. We will combine shipping for multiple items, add $1.00 for each additional knife purchased. We ship USPS 2-3 day Priority so you get your package fast! Feel free to email us with any questions because at East Tennessee Cutlery we value your concerns. Check out my other items !


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This week Highsnobiety has teamed up with with Los Angeles label GPPR (Gentleman, Philosopher, Pervert, Rebel) to offer you a chance to win $1000 USD worth of gear from their latest collection. The winners will receive a backpack, lighter case, notebook and apparel in the size of their preference.
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The contest goes live November 30, 2011. Winners will be announced December 8, 2011.


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6 Cheap Ways to Cover Windows
A designer talks window treatments for skinny budgets.
There's no doubt about it, window coverings can be very expensive and yet they also provide a high level of function for privacy, light and heat control. Depending on the location of the window, a covering of some kind may be an imperative. However, if you're on a tight budget or have lots of windows to cover, it can be a daunting task. Here are six great ways to cover your windows that are cheap and effective.
Paper accordion-pleat window shades start at under $10 per shade, are easy to install and make a great quick covering that is easily replaceable. The shades come in many colors to coordinate with your décor and different paper thicknesses offer everything from light filtering to full-on room darkening levels of light control.
- Sheer fabric panels come in many different fabrics from voile to cotton to linen. Simple sheers to lace designs are available and work well for those looking for a more formal look while staying on budget. Sheers offer great light filtering properties as well as some privacy. If you are looking for an opaque fabric, one of the easiest DIY window panels around is a twin size sheet hung on a rod. Sheets have great patterns and usually come with a hemmed header that can be converted easily to a rod pocket. Or, simply fold and run a quick stitch, or bead of glue, across the top to create the rod pocket.
- Matchstick bamboo shades are a great way to add a simple, tailored look to your windows without breaking the bank. Bamboo comes in many natural shades and allows a gentle filtered light through plus a high level of privacy during both day and night. They are usually very easy to hang – often off of a simple pair of nails in the window frame.
- Fabric tie up shades are pretty and colorful, usually washable and easy to hang. A simple pocket along the top makes for an easy rod mount and the bottom can be pulled up to the desired height by ties. These shades are not fully operable in that there is no pulley-system for easy up and down movement, but for the price (usually under $18 each), they are a cute way to dress up a window without breaking the bank.
- For those who don't care for standard window coverings but who may need a certain level of privacy, window film or translucent contact paper is a great option. Window films are readily available in home improvement stores and come in decorative "stained glass" looks or simpler "etched glass" styles. Easy to install and remove, this option works well in bathroom or basement level windows.
- Don't forget about used window treatments. Often times, a homeowner who is redecorating will be replacing older window treatments that may still be in good condition. If they were decent quality to begin with, perhaps custom, they may try to sell them or give them away. Check out local thrift stores, Salvation Army stores, flea markets or even Craigslist for some potential great deals on "hand-me down" window coverings. They may not always fit perfectly, but for rock bottom prices, you might find the perfect solution.
When dealing with how to cover one window or a whole house of windows on a budget, you don't have to sacrifice style or functionality with a little creativity and thrifty shopping.


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About the Book
Praise for Jeaniene Frost:
"Put Jeaniene Frost on your must-read list!"
—Lara Adrian
"A gutsy heroine to cheer for and a smoldering hero to die for."
—Kresley Cole
Readers can't get enough of Cat Crawfield and her sexy vampire husband Bones—especially the fans of high quality paranormal fiction who have made the novels of Charlaine Harris, Kim Harrison, Lynsay Sands, and Christine Feehan blockbuster bestsellers. With One Grave at a Time, New York Times bestselling author Jeaniene Frost gives us another spectacular supernatural adventure in her Night Huntress series—as Cat and Bones, after narrowly averting an (under)World War, must figure out how to halt the horrific killing spree of a bloodthirsty witch hunter…a sadistic killer who has already been dead for centuries.
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Product Details
- Imprint: Avon
- On Sale: 08/30/2011
- Trimsize: 4.188 in (w) x 6.75 in (h) x 0.96 in (d)
- Pages: 384
- List Price: 7.99 USD
- BISAC1: FICTION / Romance / Paranormal / Vampires
- BISAC2: FICTION / Fantasy / Urban
- BISAC3: FICTION / Romance / Fantasy


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POC Team Athlete Aaron Blunck Signature Ed.
The Fovea Aaron Blunck Signature Edition goggle has a spherical shape that ensures maximum vertical and peripheral vision, which increases the level of safety. The double lens is made of optical grade PC/CP and provides superior optical performance. The lens is anti-scratch and anti-fog treated and has ventilation slots to improve air flow and reduce fogging. The soft-coated PU frame conforms to your face and maintains its flexibility even in cold temperatures and the triple layer face foam ensures a comfortable fit. The Fovea has been developed in collaboration with POC team athlete Aaron Blunck.
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A dental filling is a treatment used to repair minimal tooth fractures, tooth decay or otherwise damaged surfaces of the teeth. Resin-based composite is the same color as tooth, and the most commonly used material. The old silver amalgam filling is never used in our office because it is unattractive, contains 50% mercury, and frequently produces cracks in teeth. Only the most up to date materials are used at the Olympia Fields Dental Associates.
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All but about 15-20 are now on DVDs in my archives. What do I do with the other several hundred? They occupy a lot of space. Many are deteriorating and have glitches on playback. I still have one good VCR but never use it anymore, DVDs only now. So do I throw them out in the trash? Give them away (who would want them)? Try to sell them (how?)? I need the storage space. I put an incredible amount of time and money into acquiring them, so they have sentimental value. Difficult decision.
No doubt many of you have had a similar conflict. How did you resolve it?
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I got rid of hundreds of VHS tapes last year through Craigslist (the free section). It took three separate postings, and dealing with an array of flakazoids and people who need to be medicated ("can you deliver them to my cabin 100 miles away from where you live?"), but they're now in the hands of others. It turns out that there are still a lot folks who actually prefer tapes to DVDs ("I can stop the tape, pull it out, play another, and put the original back in, and restart play exactly where I left off. Can't do that with a DVD"). Many seniors, in fact.
If you have CL or freecycle in your locale, then give those a shot. You'll be amazed at the response volume!
Wow- I was just about to post the exact same question! Amazing the telepathy that passes between VH members sometimes. It does seem as though the only value these tapes have is sentimental (and of course the memory of how many paychecks were doled out over the years to acquire them). I'm halfway thru my own transfer project, and beginning to seriously wonder how I'll dispose of 3000 tapes when I'm done. I always vaguely thought I could post a note on Craigs List, and some eccentric soul would come by with a U-haul to pick them up and enjoy in their big house somewhere. But thats unlikely: despite the thousands of dollars spent over the years and the breadth of the collection, the fact is no one wants VHS anymore. And since they're virtually all "personal recordings" and not commercial Hollywood tapes, I can't really advertise them on eBay or donate them to a library. At this point I'm guessing 'll probably need to pay a couple hundred $ to a recycling plant to come tow them away to a landfill once I finish the last transfer.
So sad: its like watching the last 29 years of collecting disappear into thin air . To think when I started blank TDK 120s were selling for $12.99 apiece! Thats why I shake my head at the endless number of posts from people wanting to buy the "cheapest" available media, then complaining when it doesn't work. Even at their highest current reseller prices, Taiyo Yuden 8x Premium and Verbatim 8x DataLife cost only 40 cents per disc. Compared to even current VHS prices ($1.79 for a Maxell T120), DVD-R is a bargain dream come true for old compulsive collectors like me. Although I have to admit, I have no illusions that the DVD-Rs will last as long as the tapes have: no deterioration on any of 3000 tapes over the course of 29 years in casual basement storage is pretty darned good. Whether a 40-cent disc full of semi-organic volatile dyes can be as durable as magnetic tape is something I hope I live long enough to discover .
I have accumulated well over 1000 (all VHS) and still have most of them.
I did give away about 100 each to three friends, but I can't find a home for the rest.
Charitable organizations don't want them, neither do churches or anyone involved in security/surveillance;
they all have moved to digital tape or DVD.
I suppose they'll eventually end up in a dumpster somewhere, but first, I'm going to make a few more inquiries and try to find a willing recipient.THREADKILLER !
References on File.
Originally Posted by tomlee59
Every dvd player i have ever owned retained a memory for any disc i pulled in the middle of the film.
Once after quite some period of time and several dvd's i put in a dvd and it started in the middle of the movie, i was like WTF!!!
Then i remembered, oh yeah!!! i started watching that like 2 months ago & pulled it out.
All the one's i threw out over the years were self recorded VHS tapes of tv shows.
Store bought one's took a trip to the local Goodwill.
If you have them on dvd... what's sentimental about some obsolete crappy quality bulky format
I did the same thing with my VCD's from years and years past....
When it came out on dvd, into the trash they went.
Friends did not want some crappy quality vcd disc when they could have it on dvd
I don't have nearly as many as you guys, but I've been looking at the same issue. We have a local used music store that also still handles VHS tapes, so I was going to see how many they would buy, and the ones left over will get donated to the local library. These are commercial tapes; not sure what I'll do with the few homemade tapes once I transfer them (well, the tape of our wedding I'll keep ).
As I see it, here in the USA keeping the tapes as proof that you didn't pirate the video for the DVD you made could be a good thing.
Not to mention if you still have them you have the ability to go back and recapture them with better quality later.
Originally Posted by TBoneit
Sounds like a great way to get rid of the tapes;
let the MPAA confiscate them !THREADKILLER !
References on File.
Originally Posted by jman98
A home in my small midwestern town in the USA was raided a year or two ago, which made the newspaper headlines here. I believe the guy went to prison and had to pay a big fine. Of course he was selling them (games and movies) commercially over the internet which is presumably how they caught him then decided to prosecute. I doubt it would be worth their time and effort otherwise, so those who do it privately just for themselves are probably pretty safe - as long as they don't go blabbing and bragging about it. There are probably too many in the latter category for them to do anything effective.
Common movies like Home Alone or Lethal Weapon are more or less trash. True, nobody wants those. Give them to an electronics recycle facility.
Movies not commercially available on DVD, or not available in the same version (edited, unedited, uncut, etc), are still desired. Just last night I bought a VHS tape for $5 off Amazon -- there is no DVD version of that movie. Sell or otherwise pass on these kinds of tapes, do not destroy them, collectors out there WILL want these.Want my help? Ask here! (not via PM!)
FAQs: Best Blank Discs • Best TBCs • Best VCRs for capture • Restore VHS
Originally Posted by JohnnyBob
Well, if you have any music videos / music shows / music related stuff on VHS, you can always send them to me!
I'm a huge music video collector and I'm interesting on this! Forgotten tapes from MTV for example, or other music channels you have there. Crap quality / bad quality / good quality, I don't really care. The older the better of course!
Regarding movies / etc, you can always give the tapes to your local library.La Linea by Osvaldo Cavandoli
I only had a few commercial VHS movies, but I donated them to my local library. They were in good shape and the library was glad to have them. The video section is mostly VHS, and mostly well-used tapes.
Given that most of the VHS is plastic, a good local recycle plant should take them, my local Value Village takes them a few dozen at a time.PAL/NTSC problem solver.
USED TO BE A UK Equipment owner., NOW FINISHED WITH VHS CONVERSIONS-THANKS
Originally Posted by Noahtuck
The point I was endeavoring to make is that the physical memory of a stopped tape is obviously a low-power, nonvolatile, user-friendly memory for the technically maladroit. It relies on no software, no assumptions of how a firmware engineer chose to write the underlying code, etc., and most important, relies on zero sophistication of the user. And that has an appeal for a certain not-insignificant fraction of the population. The overwhelming response to my tapes giveaway spoke to that.
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Killer Whales Too Close for Kayaker [VIDEO]
I've always said that I love being out on the ocean…as long as it's on board a good size ship or boat. You know, something that is not easily capsized by a big wave or creature like a Killer Whale.
It just so happens that a kayaker got the scare of his life when Killer Whales started popping up all around him. Think about it, the name 'Killer' is used for a reason.
If it was me, I think the water would have turned a little brown.


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Lacheys expecting second child
Nick and Vanessa Lachey are expecting their second baby, their publicists have confirmed.
The singer and VH1's Big Morning Buzz Live host first made the announcement on his Twitter account on July 15, which also marked the couple's third wedding anniversary.
He posted a photo of their almost two-year-old son, Camden, running on the beach with a heart in the sand that said: "It's a girl."
In an interview last year, 40-year-old Nick said once you have a child "everything changes, from the way you drive to the way you think to the way you act".
Vanessa Lachey, 33, last had a role on the sitcom Dads on Fox.


text: The key thing with Kyle is he needed to get into condition, lose some weight, Casey said. He did that, got into condition. And then [he has] to be a positive leader. Hes done that. Everywhere you go you have a reputation or whatever it is. And [his] was that he wasnt positive. But hes that right now. Hes doing a great job. His energy is positive. Weve just got to keep that going.If the Raptors are successful, however, Lowry will likely be a big part. Given D.J. Augustins struggles last year, the Raptors are without a sure-thing backup. Lowry is the teams best passer, and most of his shots come from the paint or behind the arc. Throw in his fearless on-court demeanor and he possesses a lot of things that the Raptors do not have in bulk things good teams need.


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Boss FS-7 Latched and Unlatched Dual Footswitch
FS-7 is a compact dual footswitch for remote control of effect pedals or amp channel switching. Two footswitches are arranged in tandem, which saves space on effects pedal boards. Two selectable modes for latch or momentary allow for control of a wide range of equipment.
- Dual footswitch in a small footprint
- Latch/momentary switching function (common to A and B)
- Durable steel chassis
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- Power via 9v adapter (power splitter included)
- Operates as momentary switch with no power*
*LEDs will not illuminate without AC power.
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FRC and Military Veterans to Hold News Conference Call in Advance of Congressional Action on 'Don't Ask, Don't Tell'
May 24, 2010, 17:58 ET from Family Research Council
FRC to Release New Polling Data, Announce Launch of New Ad Campaign
WASHINGTON, May 24 /PRNewswire-USNewswire/ -- The following advisory was issued today by the Family Research Council:
(Logo: http://www.newscom.com/cgi-bin/prnh/20080930/FRCLOGO)
WHAT: News conference call to release new national polling on homosexuals in the military and discuss FRC's new ad campaign to protect the military
Tony Perkins, President, Family Research Council
Former military leaders
HOW: Call-in at (866) 901-2585
1 p.m. EST
Tuesday, May 25, 2010
Tomorrow at 1 p.m. EST, Family Research Council President and Marine veteran Tony Perkins will be joined by retired military leaders to release new national polling data regarding the drive by President Barack Obama and Speaker of the House Nancy Pelosi (D-CA) to overturn existing law on homosexuals in the military. Perkins will also discuss FRC's grassroots outreach in the lead up to this week's expected debate and votes in Congress. Speaker Pelosi is expected to bring the issue to a floor vote, and Senate Armed Services Committee Chairman Carl Levin (D-MI) is expected to attempt adding an amendment to the Defense Authorization bill.
SOURCE Family Research Council


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'Droid to the World
Robots are the future, especially if they can pour me a beer.
By Charles P. Pierce December 30, 2007
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Dear Jeanne Dietsch: As you are the CEO of MobileRobots Inc. up there in New Hampshire, I hope I'm not being too forward in saying that I love your mechanisms. For over a decade, I have been disheartened by all those lies that the Disney people fed me in my youth about Tomorrowland. I'm 54, and I still can't go ... (Full article: 299 words)
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Canvas Draw Created Sep 05, 2012 1024 x 736px Unless noted Copyright 2012 jo_esp.
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LOVE IT! The colours, the details, the filtered sunlight coming in, the "feeling" I get when looking at it - like her joy at doing the "flips" underwater. Great!
Excellent!! collection 5/5
FANTASTIC DRAW ! very supple movements lovely coloring ! 5/5 h
so many details :) and she looks so happy..
awww, I just realized that this is another drawing, you've completed...lovely, lovely....5/5..
this is beautiful!!!, your style is so fresh and and your coloring is simply amazing 5/5
Awesome draw..love the facial expression..5H
a really great drawing she looks so free and happy amazing detail to 5h fav