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Cholangiocyte proliferation is regulated in a coordinated fashion by many neuroendocrine factors through autocrine and paracrine mechanisms. The renin-angiotensin system (RAS) is known to play a role in the activation of hepatic stellate cells and blocking the RAS attenuates hepatic fibrosis. We investigated the role of the RAS during extrahepatic cholestasis induced by bile duct ligation (BDL). In this study, we used normal and BDL rats that were treated with control, angiotensin II (ANG II), or losartan for 2 wk. In vitro studies were performed in a primary rat cholangiocyte cell line (NRIC). The expression of renin, angiotensin-converting enzyme, angiotensinogen, and angiotensin receptor type 1 was evaluated by immunohistochemistry (IHC), real-time PCR, and FACs and found to be increased in BDL compared with normal rat. The levels of ANG II were evaluated by ELISA and found to be increased in serum and conditioned media of cholangiocytes from BDL compared with normal rats. Treatment with ANG II increased biliary mass and proliferation in both normal and BDL rats. Losartan attenuated BDL-induced biliary proliferation. In vitro, ANG II stimulated NRIC proliferation via increased intracellular cAMP levels and activation of the PKA/ERK/CREB intracellular signaling pathway. ANG II stimulated a significant increase in Sirius red staining and IHC for fibronectin that was blocked by angiotensin receptor blockade. In vitro, ANG II stimulated the gene expression of collagen 1A1, fibronectin 1, and IL-6. These results indicate that cholangiocytes express a local RAS and that ANG II plays an important role in regulating biliary proliferation and fibrosis during extraheptic cholestasis.
- angiotensin
- bile duct ligation
- renin-angiotensin system
- cholangiocyte
cholangiocytes are epithelial cells that line the intrahepatic and extrahepatic bile ducts and are the target cell type in cholangiopathies such as primary biliary cirrhosis and primary sclerosing cholangitis (4, 61). Cholangiocytes are relatively mitotically dormant unless induced by certain pathological conditions such as bile duct ligation (BDL), partial hepatectomy, and bile acid feeding (23). Cholangiopathies are characterized by an imbalance between proliferation and death of cholangiocytes with proliferation playing a critical role for the maintenance of biliary mass and function during the progression of cholangiopathies (4, 61). Numerous studies have demonstrated that proliferating cholangiocytes develop neuroendocrine phenotypes and secrete and respond to a number of hormones, neuropeptides, and neurotransmitters that modulate proliferation in autocrine and paracrine mechanisms (5, 23, 44).
Angiotensin II (ANG II) is the primary bioactive component of the renin-angiotensin system (RAS) (17, 36). ANG II is produced from angiotensinogen (Ao) in two steps. First, renin cleaves Ao to form angiotensin I (ANG I), which is subsequently cleaved by angiotensin-converting enzyme (ACE) to form ANG II (17, 36). The RAS is well known for the role it plays in the regulation of blood pressure and fluid homeostasis (17, 36). It is not only important as a regulator of the cardiovascular system, but it also plays a critical role in regulating cell physiology such as inflammation, fibrosis, and tissue remodeling in a number of tissue types (7).
The biological actions of ANG II are mediated via two G protein-coupled receptors, angiotensin receptor type 1 (AT1) and angiotensin receptor type 2 (AT2) (17, 36). AT1 and AT2 have distinct downstream targets that counteract the physiological actions of each other (58). For example, activation of AT1 stimulates vasoconstriction and cell proliferation whereas AT2 mediates vasodilatation, inhibits cell growth, and activates apoptosis (58). It has been reported that ANG II induces signal transduction by activating mitogen-activated protein kinases (MAPKs) (ERK1/2, JNK, and p38 MAPK), which play key roles in cellular differentiation, proliferation, migration, and fibrosis (62, 64). ERK1/2-dependent signaling plays a key role in the regulation of cholangiocyte proliferation (23, 44).
Recent studies have demonstrated that the RAS plays a key role in hepatic fibrosis and the activation of hepatic stellate cells (HSC). In a fibrotic liver, the local RAS is induced, which plays a role in the activation of HSC (6–10, 41). Paizis and colleagues (51) demonstrated that ACE and AT1 gene expression was upregulated in response to BDL in total liver and especially in areas of active fibrosis. In liver samples from patients with hepatic fibrosis, the expression of AT1 was downregulated in hepatocytes, whereas expression was increased in HSC, vascular endothelium, and bile duct epithelium (51). Moreover, ANG II induces HSC proliferation via the activation of AT1 (8). Studies have also shown that ACE inhibitors or AT1 receptor blockers noticeably reduce hepatic fibrosis in rat (16, 30, 52, 54, 70, 75). Another recent study demonstrated that ANG II induces tumor progression and fibrosis in intrahepatic cholangiocarcinoma through interaction with HSC (49, 50). However, the direct effects of ANG II on cholangiocyte proliferation during extrahepatic cholestasis are not known. Therefore, in the present study, we evaluated the effect of exogenous ANG II on the regulation of cholangiocytes proliferation during BDL. We also explored the underlying signaling mechanism by which RAS regulates cholangiocytes proliferation.
Reagents were purchased from Sigma-Aldrich (St. Louis, MO) unless otherwise indicated. Tissue culture reagents were purchased from Invitrogen (Carlsbad, CA). The real-time PCR primers were purchased from SA Biosciences/Qiagen (Valencia, CA). The primary antibodies for Ao, renin, ACE, AT1, and AT2 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Mouse and goat IgG Vectastain ABC kits for immunohistochemistry (IHC) were purchased from Vector Laboratory (Burlingame, CA). The cyclic adenosine monophosphate (cAMP) enzyme immunoassay (EIA) was purchased from Cayman Chemical (Ann Arbor, MI). The angiotensin II (ANG II) EIA kit was purchased from Phoenix Pharmaceuticals (Burlingame, CA). The inositol phosphate-one (IP-One) ELISA kit was purchased from Cisbio US (Bedford, MA).
Animal models.
All animal experiments were performed in accordance with protocols approved by the Scott & White and Texas A&M Health Science Center Institutional Animal Care and Use Committee. Male 344 Fischer rats (150–175 g) were purchased from Charles River (Wilmington, MA) and maintained in a temperature-controlled environment (20–22°C) with 12:12-h light-dark cycles. Animals were fed ad libitum standard rat chow and had free access to drinking water. To evaluate the in vivo effect of ANG II on cholangiocyte growth, normal and BDL (immediately after surgery) rats were treated with ANG II (50 ng·kg body wt−1·min−1) (56) or 0.9% NaCl (control) by intraperitoneal implanted osmotic minipumps for 2 wk. Normal and BDL (immediately after surgery) rats were also treated with losartan (5 mg/kg body wt intraperitoneal dissolved in saline) (35) for 2 wk. Before each experimental procedure, animals were injected with pentobarbital sodium (50 mg/kg body wt ip) following the regulations of the panel on euthanasia of the American Veterinarian Medical Association.
Purification of cholangiocytes and maintenance of biliary cell lines.
Cholangiocytes were isolated by immunoaffinity separation by using a specific monoclonal antibody (IgM, kindly provided by Dr. R. Faris, Brown University, Providence, RI) that recognizes an unidentified antigen expressed by all intrahepatic rat cholangiocytes (27). The purity (98–99%) of cholangiocytes was evaluated by γ-glutamyl transpeptidase IHC (57). Cell viability (95 to 98%) was confirmed by Trypan blue exclusion. Normal rat intrahepatic cholangiocyte lines (NRIC, displaying phenotypes similar to that of freshly isolated cholangiocytes) were developed, characterized, and maintained in culture as previously described by us (3).
Evaluation of RAS expression.
The expression of the RAS components: Ao, renin, ACE, AT1, and AT2 in purified cholangiocytes was evaluated by BD Accuri C6 Flow Cytometer (BD Biosciences, San Jose, CA). Briefly, 10,000 cells were used to determine the protein expression levels. The cells were fixed by 4% paraformaldehyde and then incubated with the selected primary antibody (pAb) followed by incubation with Molecular Probes secondary antibody (Alexa Fluor 488 or 350), corresponding to the pAb species. The labeled cells were evaluated and the fluorescence-activated cell sorting (FACS) data were analyzed by using C6Flow software (BD Biosciences, San Jose, CA).
RAS component expression in purified cholangiocytes was also confirmed by real-time PCR. Total RNA was extracted by using the RNeasy mini kit (SA Biosciences/Qiagen, Valencia, CA), according to the protocol provided by manufacturer. cDNA was prepared from total RNA by using the iScript cDNA synthesis kit (Bio-Rad, Hercules, CA). SYBR Green PCR Master Mix (SA Biosciences/Qiagen) was utilized in the experimental assay with RT2 PCR rat primers designed specifically for renin (NM_012642) (11), ACE (NM_012544) (28), AT1a (NM_030985) (31, 45), AT1b (NM_031009) (31), and AT2 (NM_012494) (34) as well as for the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH, NM_017008) (65). The housekeeping gene GAPDH was used to normalize gene expression levels. Real-time PCR was performed with an ABI Prism 7900HT System (Applied Biosystems/Life Technologies, Grand Island, NY). Data were expressed as relative mRNA levels ± SE of the selected gene-to-GAPDH ratio.
The expression of renin, ACE, AT1, and AT2 was evaluated by IHC in frozen liver sections from normal and BDL rats as described (5 μm thick; n = 3 per each group of animals). Immunofluorescence for renin, ACE, AT1, and AT2 expression was performed in NRIC as described (21). Images were taken in a blinded fashion on an Olympus IX71 fluorescence microscope (Tokyo, Japan) with a DP70 digital camera.
Evaluation of proliferation of biliary and NRIC.
We evaluated the effects of in vivo administration of ANG II and losartan to normal and BDL rats on cholangiocyte proliferation by PCNA (29) and CK-19 (13) IHC as previously described (29). Proliferation assays were also performed in NRIC following in vitro treatments with ANG II (10−7 M), L-162,313 (AT1 agonist, 10−7 M) (68), CGP-42112A (AT2 agonist, 10−7 M) (15), losartan [angiotensin receptor blocker (ARB), 10−5 M] (18), H89 [cAMP-dependent protein kinase A (PKA) inhibitor, 30 μM] (22), or PD098059 (ERK inhibitor, 10 nM) (22). Cell proliferation was measured at 48 h as described (14). NRIC proliferation was also evaluated in the presence of benazepril (ACE inhibitor, 1 to 40 μM) at 48 h. NRICs were seeded (7,000 cells/well) onto 96-well plates in complete medium and allowed to adhere overnight at 37°C. Cells were then serum-starved in medium containing 0.5% FBS for 24 h, washed twice with 1 × phosphate buffered saline (PBS); 8 replicates were subsequently treated with DMSO (vehicle) or different agents. Cells were incubated for 48 h with the respective treatments, at which time 10 μl of Cell Titer 96 was added to each well. Absorbance at 490 nm was determined with a microplate reader (Spectra Max 3400, Molecular Devices, Sunnyvale, CA). Absorbance is directly correlated to the number of viable cells (43).
Evaluation of ANG II levels and evaluation of intracellular signaling mechanisms.
The concentration of ANG II in cell supernatants and plasma were measured by an ANG II EIA (Phoenix Pharmaceuticals, Burlingame, CA). NRIC (1 × 106 cells per well) were cultured in a six-well plate overnight. Then, NRICs were stimulated with vehicle (0.2% BSA) and ANG II (10−7 M) for 10 min at room temperature. cAMP levels were determined by a competitive EIA kit (Cayman) (40). Next, the effects of ANG II (10−7 M at 37°C for 90 min) on the activity of PKA were evaluated in NRIC with the PepTag assay for nonradioactive detection of PKA according to the manufacturer's protocol (Promega, Madison, WI) (6). The effect of ANG II on ERK1/2 phosphorylation was evaluated by the CASE ELISA kit (SuperArray, Frederick, MD) (67). The kit includes a complete antibody-based detection system for colorimetric quantification of the relative amount of phosphorylated protein and total ERK1/2. For the assay, NRIC were seeded into 96-well plates and were stimulated with ANG II (10−7 M), for 5, 10, 30, 60, and 180 min. In another experiment NRIC were treated with ANG II (10−7 M), ARB (10−7 M) + ANG II (10−7 M), H89 (30 μM) + ANG II (10−7 M), or ARB (10−7 M) for 10 min to evaluate the effect of inhibitors on ANG II-induced ERK1/2 phosphorylation. Detection of total and phosphorylated protein expression was determined according to the manufacturer's protocol. Finally, we measured the effect of ANG II (10−7 M at 37°C for 90 min) on the activity and phosphorylation of CREB in NRIC in the absence or presence of pretreatment with H-89 (PKA inhibitor, 10 μM). Activation and phosphorylation of CREB were evaluated by a transcription factor assay kit (Active Motif, Carlsbad, CA), which detects binding activity of phosphorylated CREB (Ser-133) (41).
Evaluation of proliferation of biliary fibrosis and profibrotic gene expression.
Liver fibrosis was evaluated by Sirius red staining for identifying interstitial collagen with red color in paraffin-embedded liver sections (4–5 μm thick, 10 different fields analyzed from each sample obtained from three different animals). Image-Pro Plus software (Media Cybernetics, Silver Spring, MD) was used for semiquantitative analysis of the staining. Briefly, a series of digital images was captured from the Sirius red-stained sections. The percentage of the fibrotic area per total liver area was calculated based on the percentage area of red color in each individual slide from each group. Fibrosis was also evaluated by IHC for fibronectin (1:100; Abcam, Cambridge, MA) as described above. Fibrotic marker expression in NRIC was also confirmed by real-time PCR as described above. RT2 PCR rat primers designed specifically for collagen 1A1 (NM_053304) (37), fibronectin 1 (FN1; NM_019143) (63), and IL-6 (NM_012589) (46) as well as for GAPDH were utilized (65). The housekeeping gene GAPDH was used to normalize gene expression levels. Real-time PCR was performed with an ABI Prism 7900HT System (Applied Biosystems/Life Technologies). Data were expressed as relative mRNA levels ± SE of the selected gene-to-GAPDH ratio.
Statistical analysis.
All data are expressed as means ± SE. Differences between groups were analyzed by the Student unpaired t-test when two groups were analyzed by Mann-Whitney U-test and Kruskal-Wallis H-test when more than two groups were analyzed, followed by an appropriate post hoc test. A value of P < 0.05 was considered significant.
Cholangiocytes express the RAS.
Cholangiocytes isolated from normal and BDL rats as well as NRIC expressed RAS components including Ao, renin, ACE, AT1a, and AT1b receptors (Fig. 1). There was a significant increase in the gene expression levels of Ao, renin, ACE, AT1a, and AT1b in cholangiocytes isolated from BDL compared with normal rats (Fig. 1A). Consistent with gene expression levels, there was a significant increase in the protein expression levels of Ao, renin, ACE, and AT1 in cholangiocytes isolated from BDL rats compared with normal rats as determined by FACS (Fig. 1B). No change in AT2 gene expression was observed during BDL (Fig. 1A). A small increase in AT2 protein expression was observed in BDL cholangiocytes as evaluated by FACS (Fig. 1B). The RAS components were also expressed in NRIC as evaluated by immunofluorescence (Fig. 1C). There was higher expression of RAS components in the bile ducts of BDL rat liver sections compared with normal rat as evaluated by IHC (Fig. 1D). Hepatocytes are also positive for the expression of RAS components as expected (76). As shown in Table 1, serum from BDL rats showed higher levels of ANG II compared with normal rats. Also, primary cultures of cholangiocytes from BDL rats secrete higher amounts of ANG II in the medium compared with normal cholangiocytes (Table 1). In addition, the normal cholangiocyte cell line, NRIC, secretes ANG II in the culture media (Table 1).
ANG II stimulates biliary proliferation.
Liver sections from the BDL rats, infused with ANG II, showed higher expression of CK-19-positive cholangiocytes per portal area compared with normal control rats (Fig. 2, A and B). Similar to the increase in the number of CK-19-positive cholangiocytes per portal area (Fig. 2B), there was also a significant increase in the number of PCNA-positive cholangiocytes per portal area (Fig. 2C). The number of both CK-19- and PCNA-positive cholangiocytes per portal area was significantly reduced in BDL rats treated with the ARB (losartan) compared with BDL + ANG II-treated rats (Fig. 2, B and C).
As shown in Fig. 3A, the AT1 agonist induced a similar increase in NRIC proliferation compared with ANG II treatment at 48 h. However, the AT2 agonist had no effect on NRIC proliferation (Fig. 3A). ANG II-induced proliferation was inhibited by ARB (losartan) (Fig. 3A). Since cholangiocytes express RAS and secrete ANG II, we evaluated the role of ANG II in the regulation of basal biliary proliferation in NRIC treated with benazepril (ACE inhibitor). Benazepril induced a significant dose-dependent reduction in NRIC proliferation at 48 and 72 h (Fig. 3B). This was also confirmed in NRIC treated with ARB, in which a significant reduction in basal proliferation was observed at 48 h.
ANG II stimulates cAMP-, PKA-, ERK1/2-, and CREB-dependent intracellular signaling mechanisms and proliferation.
In NRIC, cAMP and inositol 1,4,5-trisphosphate (IP3) levels were evaluated to begin to elucidate ANG II mediated intracellular signaling mechanisms. ANG II caused an increase in intracellular cAMP but not IP3 levels (not shown), compared with basal values (Fig. 4A). Since ANG II stimulated intracellular cAMP levels, we measured cAMP-dependent PKA activity. There was enhanced PKA activity in NRIC treated with ANG II that was inhibited by ARB (Fig. 4B). ERK1/2 has been shown to play a key role in proproliferative signaling mechanisms in cholangiocytes (20, 22, 52). A significant activation of ERK1/2 was observed in the ANG II-stimulated NRIC in a time-dependent manner (Fig. 5A). There was higher phosphorylation of ERK1/2 in NRIC within 5–10 min of ANG II treatment and the phosphorylation levels of ERK1/2 remained unchanged up to 180 min (Fig. 5A). ANG II-induced ERK phosphorylation was attenuated both by ARB (losartan) and H89 (PKA inhibitor) (Fig. 5B). This data suggests that PKA is the upstream modulator of ERK1/2, which is supported by the inhibition of ANG II-dependent proliferation by H89 and PD98059 (ERK 1/2 pathway inhibitor) (Fig. 3A). Finally, we evaluated the effects of ANG II on downstream CREB activity, which has been shown to play an important role in modulating cholangiocyte proliferation (19, 23). Stimulation with ANG II and AT1 agonist increased pCREB DNA binding activity in NRIC, which was attenuated by ARB (losartan) (Fig. 5C). Stimulation with ARB alone also resulted in decreased pCREB DNA binding activity (Fig. 5C).
ANG II stimulates biliary fibrosis and expression of profibrotic genes.
Liver sections from the BDL rats, infused with ANG II, showed significantly higher levels of Sirius red staining (i.e., fibrosis) in the portal areas compared with normal control rats (Fig. 6A). Sirius red staining was significantly reduced in BDL rats treated with the ARB (losartan) compared with BDL + ANG II-treated rats (Fig. 6A). Similar findings were observed for fibronectin staining (Fig. 6B). In vitro, ANG II stimulated increased collagen 1A1 (Col1a1), fibronectin 1 (FN1), and IL-6 gene expression in NRIC, which was blocked by ARB (Fig. 6C).
The major findings in the study relate to the expression of a local RAS by cholangiocytes and the regulation of biliary proliferation by ANG II during extrahepatic cholestasis. We found that 1) cholangiocytes express the components of the RAS and produce ANG II; 2) RAS components and ANG II levels are elevated during extrahepatic cholestasis induced by BDL; 3) administration of exogenous ANG II to normal and BDL rats increased intrahepatic bile duct mass; 4) administration of the ARB (losartan) during BDL significantly reduces intrahepatic bile duct mass; and 5) ANG II stimulates cholangiocyte proliferation via the activation of AT1 triggering cAMP-, PKA-, ERK1/2-, and pCREB-dependent signaling mechanisms. Upregulation of the RAS may play an important role in the regulation of biliary mass during extrahepatic cholestasis via autocrine and paracrine signaling mechanisms.
Previous studies have shown that RAS plays an important role in advanced liver diseases (17, 36). Several groups have evaluated the expression of RAS components in animal models of hepatic fibrosis and human tissue samples. Paizis et al. (51) demonstrated that the gene expression for ACE and AT1 were upregulated by BDL in total liver and were highly expressed in areas of active fibrosis. Similar to our findings, the biliary epithelium in tissue samples from patients with advanced liver cirrhosis expressed AT1 by IHC (59). In this study, the expression of AT1 was downregulated in hepatocytes, whereas the expression was increased in HSC, vascular endothelium, and bile duct epithelium (59).
The endocrine (or classical) RAS is known for its role in the regulation of blood pressure, electrolyte balance, and fluid homeostasis (36). Recently, studies have demonstrated the expression of all of the "classical" RAS components required for a functioning local RAS in the pancreas (38, 53). Our findings that cholangiocyte express the components necessary for the formation of ANG II, such as renin and ACE, indicate that cholangiocytes have a functional RAS. This is similar to findings observed in activated hepatic stellate cells, which have a local RAS and produce ANG II that stimulates fibrosis via activation of NADPH oxidase (77). This concept is supported by the observation in our study that cholangiocytes express the message and protein for Ao, which is the precursor for ANG II. Ao expression has also been shown in liver cells such as Kupffer cells, hepatocytes, and HSCs (42). In addition to expressing angiotensinogen, cholangiocytes also produce ANG II, which is similar to findings in activated HSCs (77). Both in vitro and in vivo, HSCs have higher expression levels of active renin and ACE and synthesis ANG II (10). Cholangiocyte production of ANG II is significantly increased during BDL in both serum and cholangiocyte supernatants. In patients with cirrhosis or chronic liver disease, there were also higher expression levels of angiotensin and plasma renin activity (69). RAS plays a key role in the alterations of portal pressure during decompensated liver cirrhosis (1). Finally, the involvement of RAS during cholestasis is supported by our finding that renin and ACE expression levels are increased in cholangiocytes during BDL. The expression of RAS in numerous liver cell types indicates that cholangiocyte proliferation may be modulated in paracrine mechanisms in addition to autocrine mechanisms. The activation of RAS in cholangiocytes may play a role in the activation of other cell types throughout the liver contributing to ANG II-induced fibrosis.
We next evaluated the effects of ANG II on biliary proliferation in both normal and cholestatic BDL rats. Chronic administration of ANG II stimulated a significant increase in biliary mass and proliferation in both normal and BDL rats, which was blocked by ARB (losartan) (AT1 inhibitor). Blockage of the RAS signaling either through ARB or ACE inhibition has been shown to attenuate fibrosis and to suppress activated HSCs and hepatic TGF-β1 expression in animal models of chronic liver injury (including CCl4 and BDL) (30, 48, 66, 72, 73, 75). In vitro, in NRIC we found that ANG II and an AT1 agonist stimulate a significant increase in proliferation, which was blocked by ARB. Interestingly, treatment of NRIC with ARB resulted in a significant reduction in basal proliferation, which supports the concept of the presence of a local RAS in cholangiocytes. We also found a significant reduction in proliferation of NRIC when treated with the ACE inhibitor benazepril. This is similar to findings reported by Li and Wanchun (39) that benazepril reduces intimal proliferation.
Next, we evaluated the intracellular signaling mechanisms regulating ANG II-induced proliferation. We found that ANG II stimulated intracellular cAMP levels but not IP3, which is in contrast to many studies (2, 12, 25). The majority of studies have show that activation of AT1 produces second messengers such as IP3, diacylglycerol, and reactive oxygen species (26). However, our data are supported by the findings that ANG II stimulates increased intracellular cAMP in glomerular epithelial cells (60). Another study has shown that that ANG II can activate cAMP formation in renal mesangial cells (55). The cAMP/PKA/ERK/CREB intracellular signaling pathway has been shown to play an important role in the regulation of biliary proliferation both in vitro and in vivo (20, 44). Therefore, we explored the downstream PKA/ERK/CREB signaling pathway related to ANG II. Similar to our previous studies, ANG II stimulated PKA activity, ERK1/2 phosphorylation, and pCREB DNA binding activity. Supporting the concept that the PKA/ERK/CREB signaling pathway plays an important role in in ANG II-stimulated biliary proliferation, inhibition of PKA or ERK1/2 signaling attenuated proliferation. The concept of the presence of a local RAS in cholangiocytes is also supported by the findings that treatment with ARB alone reduced ERK1/2 phosphorylation and pCREB DNA binding activity below basal levels.
Finally, we also evaluated the effects of chronic ANG II administration on biliary fibrosis and the expression of proproliferative/profibrotic genes. We found, similar to other studies, that ANG II stimulates a significant increase in fibrosis surrounding the portal area as well as an increase in portal fibronectin expression, which were both blocked by ARB (33, 47, 74). In a similar fashion, we found that ANG II stimulates the gene expression of collagen 1A1 and fibronectin 1 in NRIC, which was blocked by pretreatment with ARB. In vitro, we also found that ANG II stimulates a significant increase in IL-6 gene expression in cholangiocytes, which is a proproliferative cytokine that plays a key role in regulating biliary hyperplasia (71). ANG II has been shown to increase IL-6 expression levels in several cell types (24, 32). Further studies will be necessary to fully elucidate the relationship between ANG II and IL-6 on biliary proliferation and fibrosis.
The present study demonstrates that the local RAS plays an important role in the regulation of normal and cholestatic cholangiocyte proliferation and the activation of biliary fibrosis. Modulation of the RAS and the ANG II/AT1 signaling pathway in cholangiocytes may represent a novel therapeutic approach for cholangiopathies.
This work was supported by an NIH RO1 grant (DK-081442), VA CDA-2, and VA Merit Award to S. Glaser.
No conflicts of interest, financial or otherwise, are declared by the author(s).
S.A., M.K.M., A.K.M., M.U., M. Gergely, C.S., M. Guerrier, D.N., and D.E.D. performed experiments; S.A., M.K.M., A.K.M., M.U., M. Gergely, C.S., M. Guerrier, D.N., D.E.D., and S.G. analyzed data; S.A., D.E.D., and S.G. interpreted results of experiments; S.A., M.K.M., and S.G. prepared figures; S.A. and S.G. drafted manuscript; A.K.M., M. Guerrier, D.N., D.E.D., and S.G. edited and revised manuscript; S.G. conception and design of research; S.G. approved final version of manuscript.
This material is the result of work supported with resources and the use of facilities at the Central Texas Veterans Health Care System, Temple, Texas. The views presented are those of the authors and do not necessarily represent the views of the Department of Veterans Affairs. The authors thank Dr. Gianfranco Alpini for reading the manuscript and for providing the NRIC line. We also thank Anna Web for assistance with confocal imaging.
Present address for M. K. Munshi: Institute of Food and Radiation Biology, Dhaka, Bangladesh.
- Copyright © 2015 the American Physiological Society
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September 3 1968 horoscope and zodiac sign meanings.
Are you born on September 3 1968? Then you're in the right place as you can get below a lot of thought-provoking details about your horoscope profile, Virgo zodiac sign facts together with many other astrology, Chinese zodiac meanings and an engaging personal descriptors assessment and lucky features.
Horoscope and zodiac sign meanings
Just to start, these are the most often referred to astrological connotations of this date:
- The sun sign of a native born on Sep 3 1968 is Virgo. Its dates are between August 23 and September 22.
- Virgo is represented with the Maiden symbol.
- According to numerology algorithm the life path number for all born on September 3, 1968 is 9.
- The polarity is negative and it is described by attributes like self-containing and reticent, while it is considered a feminine sign.
- The element for Virgo is the Earth. Three best descriptive characteristics of natives born under this element are:
- often having a solution focused attitude
- being honest about own prejudices or egocentric tendencies
- quickly grasping patterns, principles and structures
- The modality for Virgo is Mutable. The main three characteristics of natives born under this modality are:
- deals with unknown situations very well
- likes almost every change
- very flexible
- Virgo is known as most compatible in love with:
- Scorpio
- Cancer
- Taurus
- Capricorn
- It is very well known that Virgo is least compatible with:
- Sagittarius
- Gemini
Birthday characteristics interpretation
As each birthday has its influence, so September 3 1968 carries several features of the personality and the evolution of someone born on this day. In a subjective manner are selected and evaluated 15 descriptors showing possible qualities or flaws of a person having this birthday, together with a chart that displays possible horoscope lucky features in life.
Horoscope personality descriptors chart
Little to few resemblance!
Little resemblance!
Quite descriptive!
Great resemblance!
Completely descriptive!
Quite descriptive!
No resemblance!
Sometimes descriptive!
Some resemblance!
Little resemblance!
Very good resemblance!
Great resemblance!
Good description!
No resemblance!
Rarely descriptive!
Horoscope lucky features chart
Pretty lucky!
Very lucky!
Rarely lucky!
No luck!
Great luck!
September 3 1968 health astrology
Gallstones which are basically stones in the gall bladder, crystalline concretions formed out of bile components.
Indigestion as the general term for difficult digestion caused by various factors from eating too much or eating food prepared incorrectly.
Diarrhea that can have various causes or even pathogenic agents.
Profuse sweating for no particular reason or caused by a certain agent.
September 3 1968 zodiac animal and other Chinese connotations
The Chinese zodiac offers another approach on how to interpret the influences of the birthday upon an individual's personality and attitude towards life, love, career or health. Within this analysis we will try to explain its relevance.
Zodiac animal details
- The September 3 1968 zodiac animal is the 猴 Monkey.
- The Yang Earth is the related element for the Monkey symbol.
- It is belived that 1, 7 and 8 are lucky numbers for this zodiac animal, while 2, 5 and 9 are considered unfortunate.
- Blue, golden and white are the lucky colors for this Chinese sign, while grey, red and black are considered avoidable colors.
Chinese zodiac general characteristics
- From a list that is definitely larger, these are a few general characteristics that may be representative for this Chinese symbol:
- strong person
- dignified person
- independent person
- organized person
- This zodiac animal shows some trends in terms of behavior in love which we present in this list:
- may quickly lose affection if not appreciated accordingly
- loyal
- communicative
- likeable in a relationship
- In terms of characteristics related with the social and interpersonal relationship side, this sign can be described by the following statements:
- easily manage to get admiration of others because of their great personality
- easily manage to attract new friends
- proves to be diplomatic
- proves to be ingenious
- Under this zodiac symbolism, some career related aspects which may be laid down are:
- proves to be results oriented
- learns quickly new steps, information or rules
- prefers learning through practice rather than reading
- proves to be specialist in own working area
Chinese zodiac compatibilities
- The Monkey and any of the following zodiac animals can have a successful relationship:
- Snake
- Rat
- Dragon
- A relationship between the Monkey and the following symbols can evolve nicely at the end:
- Rooster
- Goat
- Monkey
- Pig
- Ox
- Horse
- A relationship between Monkey and any of these signs it is unlikely to be one of success:
- Tiger
- Dog
- Rabbit
Chinese zodiac career
If we look at its characteristics a few great careers for this zodiac animal are:
- investment officer
- operations officer
- business analyst
- researcher
Chinese zodiac health
A few things related to health should be in this symbol's attention:
- should avoid any vicies
- should try avoiding worrying for no reason
- should try to keep a proper diet plan
- has an actyive lifestyle which is positive
Famous people born with the same zodiac animal
These are a few famous people born in the Monkey year:
- Leonardo da Vinci
- George Gordon Byron
- Demi Lovato
- Bette Davis
This date's ephemeris
The ephemeris positions for this birthday are:
Sidereal time: 22:48:46 UTC
Sun in Virgo at 10° 33'. Moon was in Capricorn at 20° 26'. Mercury in Libra at 01° 55'. Venus was in Libra at 00° 53'. Mars in Leo at 18° 08'.
Jupiter was in Virgo at 15° 05'. Saturn in Aries at 24° 56'. Uranus was in Virgo at 28° 24'. Neptun in Scorpio at 23° 59'. Pluto was in Virgo at 22° 13'.
Other astrology & horoscope facts
On September 3 1968 was a Tuesday.
The soul number that rules the September 3, 1968 date is 3.
The celestial longitude interval related to Virgo is 150° to 180°.

subset: commoncrawl


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Must See! This is an adorable home with a great lay out that makes the most of all space available . Well maintained inside and out. Beautifully tiled floors are paired with high ceilings, lots of natural light and charming French doors that lead out from the kitchen . Enjoy relaxing on the deck out back. This home is move in ready ! Pictures coming soon Buyer or buyers agent to verify all information


text: Triple decker burnout.(video in post)Singhjr962/15/14 6:35pm31EditPromoteDismissUndismissHideShare to KinjaGo to permalink So there's a truck, an ute, and a motorcycle, all stacked on each other. These guy's are either properly crazy, or insanely brilliant. I'm going to say brilliant.skip to about the :26 second mark for instant burn out gratification.Reply3 repliesLeave a reply


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Hydro Aluminum Rockledge Inc
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Rio Olympics might be a mess, but Tokyo 2020 has some cool high-tech plans
Driverless taxis for athletes and holograms for info.
Thu, Jul 07 2016
Clinton takes big step toward easing tensions with Sanders
With new college affordability plan, the presumptive Democratic nominee won praise from rival.
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4 servings, 1-2/3 cups (400 mL) each
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Substitute baby spinach leaves for the bok choy.
Top with sliced green onions and toasted sesame seed just before serving.
Help your friends and family eat right with this delicious stir-fry. As a bonus, the bok choy provides an excellent source of both vitamins A and C.
* Nutrition information is estimated based on the ingredients and cooking instructions as described in each recipe and is intended to be used for informational purposes only. Please note that nutrition details may vary based on methods of preparation, origin and freshness of ingredients used.
Added mushrooms because we like them and used spinach. Very tasty. Will make again.
Very good; used broccoli rather than bok choy as it was what I had on hand.
I halved the recipe as there is only two of us. I also used baby spinach instead of bok choy as that is what I had on hand, added a few extra snap peas and some mushrooms it was great.


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Welcome to Librarium Online!
Join our community of 80,000+ members and take part in the number one resource for Warhammer and Warhammer 40K discussion!
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So, I took first in my City tournament to qualify for Regional competition in Utah, for the second round of the 'Ardboy tournament. Props to the Hydra, and to Kroot Mercenaries. (USE THEM)
Game 1: Tyranids
Played just like a Recon, except for a Ordnance blast every turn due to "Firestorm". That really helped pop off some Hormagaunts. The TMCs were then torn to bits by my Plasma Rifles.
Game 2: Death Guard
WOW tough game (Cleanse/Take and Hold hybrid). All troop choices in Reserve.. it was tough to keep him out of my deployment zone, and to keep Scoring units in the other Table Quarters. Fortunately, I destabilized the Great Unclean One, and the Kroot Headhunters sprayed it down with acid.. picture Mortal Kombat:
Game 3: Tau
Hilarious Secure and Control. 800 point O'Shovah + 9 Crisis Bodyguard unit gets charged by my Chaos Lord. After felling 1 Crisis, the whole unit flees the battle. After that.. the Kroot Mercs killed 12 Fire Warriors, 3 Broadside, Sniper Drone team, and Pirahna. Strong finish, for sure.
Did anyone else play the 'Ardboy? Did anyone qualify for Regionals? I'll be in Clearfield, Utah on October 13th, and I was hoping to see some Librarium users there.
Here's the list I used: HERE!


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Townhouse/condo has 3 beds, 2 baths, 1,426 sq. ft., 1426 sq. ft. lot size, was built in 1958 and is pet friendly. Offered as for sale by owner with a price of $2,134.
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This property is For Sale by Owner or FSBO , which can offer significant advantages to home buyers. Even though a homeowner chooses FSBO to save money on real estate fees, those savings can be passed on to the buyer as a lower purchase price. Some FSBO homeowners may also offer flexible financing options, including Owner Financing (a good alternative to a bank loan) or Rent to Own. The Rent to Own option would allow you to rent this home for a period of time with an option to purchase it for a predetermined price after the rental period. This...
gives you time to build up your credit and accumulate your down payment, all while living in the home of your dreams.
Townhouse located at Inglewood in North Andover, MA 01845 has 3 beds, 2 baths, 1,426 square feet and available to rent for $2,134. Inglewood was listed on RentalSource 35 days ago by RentBeforeOwning. Inglewood is in zip code 01845 which is located in North Andover, MA. The rental price of $2,134 is 34% higher than the average rental price of $1,601 for townhome rentals in North Andover, MA 01845.Report listing | Visit website
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Local perfusion of the dorsal root ganglion (DRG) with tumor necrosis factor α (TNF-α) in rats induces cutaneous hypersensitivity to mechanical stimuli. Thus we investigated the cellular mechanisms of TNF-α-induced mechanical hyperalgesia. The L4and L5 DRGs with the sciatic nerves attached were excised from rats for in vitro dorsal root microfilament recording. After baseline recording for 15 min, TNF-α (0.001, 0.01, 0.1, or 1 ng/ml) was applied to the DRG for 15 min, followed by washout for at least 30 min. Alternatively, H-89 or Rp-cAMPS, two specific cAMP-dependent protein kinase (PKA) inhibitors, was added to the perfusion solution for 15 min prior to TNF-α application. TNF-α (1 ng/ml) induced neuronal discharges in 67% (14/21) of C fibers and 27% (4/15) of Aβ fibers when applied topically to the DRG. Acute TNF-α application not only evoked discharges in silent fibers, but also enhanced ongoing activity of spontaneously active fibers and increased neuronal sensitivity to electrical stimulation of the peripheral nerves. H-89 (10 μM) and Rp-cAMPS (100 μM) each completely blocked the TNF-α-evoked response in most C and Aβ fibers tested but did not affect fiber conductivity. Our results demonstrates that exogenous inflammatory cytokines such as TNF-α can elicit a PKA-dependent response in sensory neurons and thus strongly suggest that endogenous TNF-α may contribute to the development of certain pathological pain states.
Proinflammatory cytokines such as tumor necrosis factor α (TNF-α) are involved in the development of inflammatory and neuropathic pain behaviors. The level of TNF-α in inflamed tissue was significantly increased after intradermal injection of endotoxin (Kanaan et al. 1998). Conversely, the hyperalgesic effect induced by carrageenan was limited by an antiserum to endogenous TNF-α (Cunha et al. 1992). In neuropathic animal models, it has been shown that TNF-α mediates thermal as well as mechanical hyperalgesia after nerve injury (George et al. 2000; Ignatowski et al. 1999; Sommer et al. 1998a,b; Wagner and Myers 1996). In previous work in this laboratory, TNF-α induced mechanical hyperalgesia when administrated locally to the normal dorsal root ganglion (DRG), and enhanced ongoing hyperalgesia when deposited at the chronically compressed DRG (Homma et al. 2002). However, little is known about the neurological and cellular mechanisms underlying these effects.
In rat peripheral nervous system, TNF-α evokes action potentials in nociceptive neurons when applied topically to peripheral axons in vivo (Junger and Sorkin 2000; Sorkin et al. 1997). Nicol et al. (1997) found that chronic treatment of the DRG cells with TNF-α enhanced capsaicin sensitivity of isolated sensory neurons. The enhanced sensitivity is likely to be mediated by the neuronal production of prostaglandins, as treating the cells with cyclo-oxygenase-2 (COX) inhibitors blocked the TNF-α-induced sensitivity enhancement. Some of these data can be used to explain, at least partially, the hyperalgesic effects of TNF-α when administered acutely to the nerve trunk (Sorkin and Doom 2000; Wagner and Myers 1996) or when injected subcutaneously into the rat hindpaw (Perkins and Kelly 1994; Woolf et al. 1997).
The role of cyclic AMP-dependent protein kinase (protein kinase A, PKA) pathway in the maintenance of inflammatory pain was demonstrated recently by the finding that PKA inhibitors reduced hyperalgesia induced by hyperalgesic agents (e.g., prostaglandin E2, and purine) (Aley and Levine 1999). Furthermore, a role for PKA in mediating nerve injury-induced neuropathic pain was indicated in a study demonstrating that intrathecal spinal delivery of H-89 (a specific PKA inhibitor) reduced the hyperalgesia resulting from a unilateral tight ligation of L5 and L6 spinal nerves (Hua et al. 1999).
In the present study, using extracellular electrophysiological techniques, we examined the effects of acute application of TNF-α on the excitability of DRG neurons. The intracellular transduction cascade, in particular the role of PKA pathway in mediating the interaction between TNF-α and neuronal excitability, was also investigated.
Extracellular electrophysiological recording
Male Sprague-Dawley rats (150–200 g, n= 54) were anesthetized with pentobarbital sodium (40 mg/kg ip). The L4 and L5 ganglia with attached dorsal roots (length: about 2 cm) and sciatic nerve (length: about 3 cm) were dissected and placed in a recording chamber (Zhang et al. 1997). The DRG was perfused at a rate of 5 ml/min with oxygenated artificial cerebrospinal fluid (ACSF) containing (in mM): 130 NaCl, 3.5 KCl, 1.25 NaH2PO4, 24 NaHCO3, 10 dextrose, 1.2 MgCl2, and 1.2 CaCl2(pH = 7.3). The perfusion solution was heated to maintain the bath temperature at 37°C. The dorsal root was led out of this chamber into an adjacent mineral oil-filled chamber where microfilament recordings were performed. The spinal/sciatic nerve (length: about 3 cm) was contained in an adjacent chamber with mineral oil and placed in contact with a bipolar stimulating electrode. Each chamber was separated by petroleum jelly (Vaseline) to prevent the solutions from different chambers from intermixing.
Dorsal root microfilaments were teased apart under a dissecting microscope. The proximal end of a dissected microfilament was placed on a fine silver electrode for single fiber recording. The discharges of single fibers were displayed on a digital oscilloscope and collected via Spike 2 data-acquisition system (Cambridge Electronic Design, Cambridge, UK) on a Pentium III PC. The conduction velocity of each fiber was obtained via electrical stimulation delivered to the sciatic nerve.
Drug preparation and application
Recombinant human TNF-α (R and D Systems, Minneapolis, MN) was dissolved in 0.1% bovine serum albumin (BSA) in buffered saline to a concentration of 100 ng/ml and stored at −80°C in 10 μl aliquots for later use. The H-89 (Sigma Chemicals, St. Louis, MO) was dissolved in methanol at 0.5 mM and diluted to final concentration of 10 μM (containing methanol 0.0025%, vol/vol) prior to recordings. Rp-cAMPS (Sigma Chemicals) was dissolved in distilled water and diluted to 100 μM immediately prior to drug application. The pH for all drugs used in the present study was 7.3.
To determine whether the acute application of exogenous proinflammatory cytokines directly evokes action potentials in the somata of lumbar ganglia, TNF-α (0.001, 0.01, 0.1, or 1 ng/ml) was applied to the DRG for 15 min after a 15-min baseline recording. Alternatively, either Rp-cAMPS or H-89 was applied to the DRG before the TNF-α application to test the involvement of the PKA pathway. Briefly, H-89 or Rp-cAMPS was applied to the DRG for 15 min, followed by Rp-cAMPS or H-89 plus TNF-α for another 15 min. In some experiments, after 30–40 min washout with ACSF, the same dose of TNF-α (containing the same vehicle as the first application) was once again applied to the DRG for 15 min.
Data acquisition and analyses
With the employment of Spike 2 data-acquisition system, we could examine up to three C or Aβ fibers with different amplitudes during each trial. A mean basal ("control") discharge rate was computed as the mean number of spikes/s (±SE) for 15 min before delivery of TNF-α. Spontaneously active was defined as fibers with any number of spikes in 5-min interval. For each fiber, the maximal or peak effect of TNF-α on the fiber's activity was defined as the discharge rate in the 5-min interval (spikes · s−1 · /5 min−1) following drug administration that exhibited the greatest increase from the peak basal rate. Student's two-tailed t-test was used to compare the response latencies between silent and spontaneously active fibers. The criterion for significance was P < 0.05.
Silent vs. spontaneously active C and Aβ fibers
Myelinated Aβ and unmyelinated C fibers were activated using an in vitro nerve-DRG-dorsal root preparation by electrical stimulation applied to the sciatic nerve in 54 rats. A total of 148 fibers were activated and identified as C fibers with conduction velocity ranging between 0.48 and 1.79 m/s (mean conduction velocity: 0.9 8 ± 0.05 m/s). Eight-five of the 148 C fibers (74 silent and 11 spontaneously active) could be isolated using the Spike 2 acquisition program and thus were used for the present electrophysiological study. The discharge rate was low and extremely variable, ranging between 1 and 150 spikes in a 5-min interval. Eight of 11 fibers had less than 12 spikes in a 5-min interval. The mean discharge rate over 15 min for the 11 spontaneously active C fibers was 0.1 1 ± 0.0 4 spikes · s−1 · 15 min−1. The discharge pattern was irregular for all but one fiber, which exhibited a short burst-type discharge. Of 34 Aβ fibers used in this study, 30 were silent and 4 were spontaneously active with conduction velocity >15 m/s (Harper and Lawson 1985; Villiere and McLachlan 1996) and a low discharge rate of 0.03 ± 0.02 spikes · s−1 · 15 min−1 (n = 4).
Acute application of TNF-α evoked responses in DRG neurons with slow-conducting C fibers and fast-conducting myelinated Aβ fibers
A total of 20 C fibers were tested with 1 ng/ml of TNF-α, 7 fibers were spontaneously active and 13 fibers were initially quiescent. TNF-α elicited discharges in 7 of 13 silent fibers, enhanced the firing rate by at least 30% in 6 of the 7 spontaneously active fibers (Fig. 1) and suppressed the firing rate in 1 spontaneously active fiber. The average latency for TNF-α to evoke discharges in initially quiescent fibers was 21 ± 7 min (range, 5–43 min). In four fibers, TNF-α evoked discharges within 15 min of drug application. The remaining fibers started responding after the beginning of the washout. The latency between TNF-α application and enhancement of the spontaneously active fibers (17 ± 5 min) was shorter but not significantly different from that of silent fibers (P > 0.05, Student'st-test). The evoked activity usually lasted for more than 30 min in response to a 15-min TNF-α application. After a 30-min washout, 50% of the tested fibers returned to basal level, but the remaining fibers did not recover for at least 1 h after TNF-α application. However, in two initially quiescent fibers, the evoked response lasted for only 10–20 min.
TNF-α was tested twice in one spontaneously active fiber. The first TNF-α application increased peak discharge rate from 0.02 to 0.27 spikes/s, with a latency of 2.6 min. After 30-min washout, a subsequent application of TNF-α at the same dose evoked a weaker response (peak discharge rate of 0.07 spikes/s) and a longer latency (30 min).
Three lower doses of TNF-α (0.001, 0.01, and 0.1 ng/ml) were also tested. None of six fibers that were tested with 0.001 ng/ml of TNF-α responded. Of the seven fibers (including 3 fibers that did not respond to 0.001 ng/ml) that were tested with 0.01 ng/ml, only two responded (including 1 that did not respond to 0.001 ng/ml) and had a latency of 30 and 44 min, respectively. Of 19 fibers (18 silent, 1 spontaneously active) treated with 0.1 ng/ml, 9 fibers exhibited enhanced firing with an average response latency of 23 ± 4 min. Only two of these fibers recovered after 30-min washout with ACSF alone. Of the 19 fibers tested with 0.1 ng/ml of TNF-α, 8 were also tested once with TNF-α at 0.01 ng/ml. No higher doses than 0.1 ng/ml of TNF-α were tested prior to the application of 0.1 ng/ml in any of the 19 fibers. Although variable doses of TNF-α elicited different response latencies, the peak discharge rates were not significantly different (P > 0.05, ANOVA; Fig.2).
TNF-α not only evoked discharges in silent C fibers but also enhanced neuronal sensitivity to electrical stimulation of the peripheral nerves. In 32 C fibers, a single stimulation of the nerve only evoked a single action potential prior to TNF-α application. However, 15 min after TNF-α application (1 ng/ml), the same current pulse evoked a long-lasting burst in 7 of 32 fibers (22%; Fig.3). The duration of electrically evoked firing varied among different fibers and ranged from 5 to 30 s, similar to that found in some DRG cells with chronic compression injury as reported previously (Zhang et al. 1999).
TNF-α (1 ng/ml) also elicited discharges in 4 of 15 (27%) quiescent Aβ fibers (Fig. 4). The average response latency was 24 ± 8 min. Only one of the four fibers that responded to TNF-α returned to its basal level after the 45-min washout. Two of nine Aβ-fibers (22%) responded to 0.1 ng/ml and one of five fibers (20%) responded to 0.01 ng/ml of TNF-α.
In a separate experiment, to eliminate the possibility that TNF-α-induced response could result from the instability of the fibers tested over a long period of time, the baseline activity from a total of nine silent and four spontaneously active C fibers was recorded for 60 min. No activity was recorded from any silent C fibers during the 60 min ACSF perfusion. Furthermore, no significant change (more than 30% over basal rate) was observed in any of the four spontaneously active C fibers.
Excitatory effect of TNF-α was blocked by H-89 or Rp-cAMPS
A total of eight C fibers (2 spontaneously active) were treated with H-89 (10 μM) for 15 min, followed by application of TNF-α (1 ng/ml) plus H-89 for another 15 min. None of the eight fibers responded to TNF-α. However, after the washout with ACSF for 30–60 min, TNF-α (1 ng/ml) induced responses in six of the eight fibers that had not responded to TNF-α plus H-89 (Fig.5).
In five silent Aβ fibers, TNF-α failed to evoke responses when applied to the DRG together with H-89. After washout with ACSF alone for 30 min, a second application of TNF-α without H-89 evoked responses in four of five fibers tested. The average response latency was 10 ± 3 min, and mean peak discharge rate was 0.34 ± 0.25 spikes · s−1 · 5 min−1. None of the four fibers recovered from TNF-α-induced response after the 30-min washout period.
In a separate experiment, six C fibers were treated with Rp-cAMPS (100 μM) for 15 min, followed by Rp-cAMPS and TNF-α (1 ng/ml) for another 15 min. No neuronal discharge was elicited in five of six fibers tested while a weak and transient response was induced in only one fiber. After 30-min washout with ACSF, TNF-α alone was added to the perfusion solution, and a response was elicited in four of six fibers tested, including the fiber that had responded previously. For this fiber, TNF-α alone evoked a much stronger response than did TNF-α plus Rp-cAMPS in the previous test. Alternatively, in three C fibers, TNF-α (1 ng/ml), instead of Rp-cAMPS, was first applied to the DRG for 15 min prior to the treatment with Rp-cAMPS. TNF-α alone evoked a robust discharge in all three fibers. However, the second application of TNF-α plus Rp-cAMPS failed to evoke any action potentials in one of three fibers and evoked a transient weak response in the remaining two C fibers tested (Fig.6).
Although PKA inhibitors (H-89 or Rp-cAMPS) blocked the acute TNF-α-elicited responses, the fiber conductivity, as examined by electrical stimuli of the peripheral nerve, was not interrupted in any fibers during or after the administration of the PKA inhibitors.
The present study demonstrated the ability of TNF-α to evoke discharges in DRG neurons with myelinated or unmyelinated axons. This effect likely is mediated by a PKA pathway as pretreatment of the ganglia with specific PKA inhibitors blocked TNF-α-induced responses.
A suppressive effect was only observed in a minority of spontaneously active C fibers even when higher dose of TNF-α (1 ng/ml) was employed. In previous studies, it was found that the ongoing activity was decreased in the majority of spontaneously active C fibers in response to higher doses of TNF-α (>0.05 ng/ml) applied topically to the nerve trunk (Sorkin et al. 1997). It is possible that the sensitivities to exogenous TNF-α are different between nerve fibers and cell bodies. The use of human, instead of rat, TNF-α in the present study might be another contributory factor to the discrepancy in TNF-α sensitivity between our and other studies. Although the amino acid sequence homology of human and rat TNF-α is 89% (Kwon 1993), it is expected that human TNF-α might have lower affinity to rat receptors.
Most fibers responded to TNF-α after exposure of at least 10-min TNF-α duration. The relatively long response latency suggests that an intracellular signal transduction pathway may have been involved in TNF-α-evoked responses. A similar response pattern has been observed recently. In a study exploring the effect of PKA on the modulation of spontaneous activity, most fibers responded to PKA activators after 15 min of topical application (Hu et al. 2001). The variability in response latency and recovering time might be due to the time for the drug to reach the somata that are located at different levels beneath the surface of the ganglion. The discrepancy in response latency between current results and results reported previously (Sorkin et al. 1997) may lie in the use of TNF-α at different pH. The use of different types of TNF-α (rat vs. human) in two studies might be another cause of the discrepancy in response latency.
The firing frequency elicited by TNF-α is greater in quiescent fibers than in spontaneously active fibers. The differences might be explained by partial phosphorylation of certain ion channels contributing to the generation of spontaneous activity. Earlier studies have shown that spontaneous activity of DRG neurons can be increased by protein phosphatase inhibitors such as okadaic acid (Hu et al. 2001), suggesting that ion channel phosphorylation may be involved in the signal transduction pathways that modulate spontaneous activity. These channels are likely potassium channels as demonstrated in earlier studies on TNF-α (Diem et al. 2001). A partial phosphorylation might have caused inactivation of certain numbers of potassium channels and decreased potassium conductance, which resulted in reduced magnitude of responses of spontaneously active fibers to any further drug applications.
Subsequent experiments with specific PKA inhibitors, H-89 and Rp-cAMPS, demonstrated that TNF-α-induced responses are PKA dependent. These results agree with previous reports that G-protein-mediated activation of PKA is one of the several signal transduction pathways that can be activated by TNF-α (Pan et al. 1997). It has been assumed that sensitization of nociceptors is due to increased concentrations of cAMP/Ca2+ in the sensory neurons (Cui and Nicol 1995; Ferreira 1993). This hypothesis is supported by results from a recent study in which blocking the PKA pathway with H-89 or Rp-cAMPS suppressed spontaneous activity of Aβ and Aδ fibers (C fibers were not tested) originating in the ganglia subjected to a previous chronic compression injury. Increasing intracellular cAMP level, on the other hand, enhanced ongoing spontaneous activity of DRG neurons (Hu et al. 2001). Thus it is likely that the excitatory effects of acute application of TNF-α on the DRG somata may have resulted from elevated intracellular cAMP level (Ebadi et al. 1997). The latencies for TNF-α to evoke neuronal responses were shorter in both A and C fibers tested previously with H-89 plus TNF-α. This suggests that earlier application of TNF-α may have partially sensitized the tested fibers, although no discharges were evoked in the presence of H-89.
The present study has demonstrated a PKA-mediated TNF-α response, a mechanism that is similar to prostaglandins E2(PGE2)-induced sensitization of DRG neurons (Cui and Nicol 1995; Evans et al. 1999;Lopshire and Nicol 1998). It is possible that topical application of TNF-α may have caused PGE2release through activation of COX-2 pathway as suggested previously (Nicol et al. 1997). However, a more extensive study is needed to determine if acute TNF-α application-evoked responses can be blocked by specific COX-2 inhibitors.
Clinically, lumbar ganglia and the adjacent dorsal roots are exposed to inflammatory cytokines (e.g., TNF-α) released from a ruptured lumbar disk (Kang et al. 1996). Our findings that TNF-α may evoke discharges in DRG neurons with myelinated as well as unmyelinated axons strongly suggest that, in addition to cutaneous hyperalgesia in inflammatory and neuropathic animal models, inflammatory cytokines also contribute to the initiation and maintenance of low back pain in patients with the preceding pathological conditions.
This work was supported by National Institute of Neurological Disorders and Stroke Grant R01NS-39568A.
Address for reprint requests: J.-M. Zhang, Dept. of Anesthesiology, Slot 515, University of Arkansas for Medical Sciences, 4301 W. Markham St., Little Rock, AR 72205 (E-mail:ZhangJunming{at}uams.edu).
- Copyright © 2002 The American Physiological Society


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- 09-23-2015, 05:36 AM Thread Author #1iMore Nanite
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Proximity Sensors with iOS9
I just upgraded to iOS9 and the screen not litting up when the phone is face down due to the proximity sensors in the phone is been advertised as one of the key features to upgrade. However in my face the phone does lit on every notification even if it is face down. Why is that?
Also I notice iOS9 is a little sluggish on iPhone 5 where the system lags before doing anything. Anyone else had that issue?
Someone has confirmed that this works fine on iPhone 6 Plus. I wanted to know if this works on iPhone 5?
- 09-23-2015, 02:34 PM #2iMore Genius
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Farrah Fawcett contentVisited: 11142
165 pics / nude
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5 vids / nude
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Farrah Fawcett
Birth Date: 02.02.1947.
Birth Place: Corpus Christi, Texas, U.S..
Occupation: Actress
Nationality: American
Appear in: Blonde Vintage
System rating: 4.65
Content: 165 pictures and 5 videos
More info [+]
Farrah Fawcett was an American actress and artist. A multiple Golden Globe and Emmy Award nominee, Fawcett rose to international fame when she first appeared as private investigator Jill Munroe in the first season of the TV series Charlie's Angels, in 1976.Fawcett was a sex symbol whose iconic poster, released the same year Charlie's Angels premiered, broke sales records, making her an international pop culture icon. Her hair style was emulated by millions of young women in the 1970s and early 1980s
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Real news. Real funny.
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But being so obsessed about a goal can have its little drawbacks, as (a) it affects negatively your relationship with other people and (b) you may confuse "motivation" with "effort". That's what happened to me.
Since the first day of class was approaching, I anxiousy decided to google "how to quickly revise japanese" and stumbled upon a site that began like this:
"Hello. I suppose you ended up in this site because you probably googled 'how to quickly revise japanese' didn't you? Well, here I present you with some methods to speed up revision, but you have to be committed. Before I describe to you each method, ask yourself how much determined you are to learn Japanese. If the answer is "very determined", then don't waste a good chance. For example, you have a couple of spare minutes and you're scrolling down your facebook page? Use them to revise japanese numbers instead. You've washed your teeth and are going to bed? Make a quick revision of Japanese months before sleep. You are bored and want to watch youtube videos for fun? Why not watch an episode of a j-drama/anime to impove your listening instead? Any moment you could be doing something not-so-important, choose to revise, in any way. That's the most important thing."
I was impressed and decided to follow the advice.
A few days later, I caught myself panicking with thoughts like:
O__O I am trying. I really am.
....and I am going to mentally throw up.
My mind is rejecting knowledge, refusing to memorise, after being at work all day and using the breaks in between to revise. I don't know how much I'll last. Burnout. Overdoing it. Too much pressure. Exhaustion. That's what I've started to feel, and it's only October, classes haven't even started yet, and when they do it will last till June. No way. But I want it too much. My will alone pushes the body to keep going, but it's nauseating and I'm starting not to feel so well... DX Ok, must take a break!!! Happy thoughts, err....I wish I took the day off, walked around, went shopping... gahh relax, calm down, self. Ok, this Sunday no power in the world will manage to make me get up for work. I'll just enjoy the day to the fullest; water my little plant, hum a tune, cook something nice, do non-mental chores, make the house smell clean, have some tea... XD Can't wait to smell the freshly-ironed clothes and feel the sense of coziness again. Allllll day to myself, or with my family. Pheww, it's working, I feel better already...Nope, I'm lying. My throat is tightening.
*deep breaths*
*meditates- Ohmmm...*
Ironically, the only moment of the day I was able to relax was when I went to teach. Finally, some peace. Since it requires the maximum of my attention, distractions are left outside. Time stops in that class and I don't even remember who I am or notice how the sun is slowly setting; I'm only aware that I'm teaching. Trust me, I've temporarily fogotten whole disastrous fights with boyfriend in that room. It does miracles.
When I came out I was feeling calmer. I never expected to like teaching so much, to feel so...safe doing it. So now I understand that while it's commendable to try hard for your dreams, you don't have to kill yourself in the process. I guess even if I don't manage to revise everything, the rest will come on its own, once we start.
*...revises verb "start" in japanese"
It was only when I uttered my worries through tears, bringing them out in the open, that I finally realised how deep they ran inside of me. No way, unbelievable. I couldn't be that much insecure a person. I couldn't be the girl who was breaking apart from low self-esteem in front of her boyfriend. Finally, after poking inside like an expert psychologist, past the hurt, I was able to find those core beliefs that hadn't even formed thoughts yet. There they were. They rang so true.
They caused me feelings of misery, self-pity, weakness... and suddenly I felt so uneasy. I wanted to throw up, to reject it, /this is not me/, I got sick of it.
The next day, I had an argument with J. and got stubborn. Instead of driving to meet him, I made a decision on the spot and took another turn. A change.
Trying not to think much about my feminility, my vanity -all the times spent brushing my long hair in front of the mirror after showering- I pushed open the hairdresser's door and went in for a drastic cut.
I didn't care if it turned out pixie-like, if I ended up looking like a boy, a tomboy, a lesbian or I-don't-know-what and he wouldn't approve of it. It would make me feel "me". My true self has short hair, I think. "Long" was no longer me, it felt foreign, associated with years of weakness piling up and I couldn't stand a minute more wearing it - I wanted it gone, even if I had to do it myself! (A sort of borderline hysteria had got over me, I wonder.)
The hairdresser gave me a weird look of 'why would you want to cut this beautiful hair' but nevertheless brought me the picture catalogue to chose from and did her best to guide me through it.
Difficult choice, normally I would panic. I hadn't had time to think what would suit me, but I knew what I didn't want: I did NOT want straight bangs on the forehead like a monk; NOT a bob cut that would make me look like I was wearing a helmet; NOT parted in the middle like Jesus, but to the side insead. I went for locks of uneven lenghth, volume at the top, layers around my face.
When she gave the first cut, she paused and looked at me through the mirror. 'Perhaps you would like to keep it?' she said, offering me the locks. I refused, surprising myself. I had expected some sort of sentimental attachment, but found only relief. No, I didn't want to keep it. And however it turned out, I was glad to have chosen it. I wasn't regretting it.
I kept a genuine smile thoughout the whole process and by the end, both her and I were ethusiastic with the result. Now, that was me. She said she was a little jealous, actually. Maybe because she witnessed a change. After the cut, I went to a clothes shop nearby and chose a nice, sleeveless top of intense blue, like the clear sky, which replaced my faded brick-red T-shirt. Farewell, people-pleaser. Welcome, new, hot, free girl.
It was like this that I went to meet him outside his house. I can now smile confidently. He has no idea what's coming for him. He's got to learn me again.
- Bought a sewing machine and learnt how to shorten curtains. (Not physical, but still proud!)
- Sunday, July 3rd : Hiking with friend, covered 7 klm.
- Monday, July 4th : Volunteered to take stray dogs for a walk. Spent 1.5 hours walking, while they pulled. Overcame my fear of being attacked by that one who jumps on me every time.
- Tuesday, July 5th: Aside from a looong walk for coffee...I feel restless. Edit: asked a nearby karate school about their prices and schedule...Plan to join them in September!
- Wednesday, July 6th: Tried modern dance lesson for 3 hours. I hurt everywhere....I pushed myself to the point that lifting a leg to wear my pyjama pants was nearly impossible.
Edit: A friend just decided to join me with the group. We're hiking this Sunday again!
Edit 2 : Glad we went! We were able to snap this picture :D
List of Concerts J. Has Been to
.__. ok, I get it. I don't even know half of these guys... but that's a lot of ticket money.
Found this on a blog and just had to repost it:
And now, if you will, a revised version of the Rifleman's Creed, for us writer-types –
This is my book. There are none like it, because this one is mine.
My book is my best friend. It is my life. I must master it as I must master my life.
My book, without me, is useless. Without my book, I am useless. I must write my tale true. I must shoot straighter than my fear who is trying to kill me. I must kill my fear before my fear kills my story.
My book and I know that what counts is not what others have done, what sales we make, what tweets I have twotted. We know that it is my heart that counts.
My book is a living document, because it is my life. I will learn it as it is my kin. I will learn its weaknesses, its strengths, its characters and plots and themes. I will put my heartsblood into the book and it will put its heartsblood into me as we become part of each other.
Before the Muse that I have shackled to the radiator in my office, I swear this creed. My book and I are the representatives of who I am. We are the masters of our fear. We are the ink-stained fools who press our fingerprints into the page for all to see. We are story and story-teller, one and the same. We are the gods of this place.
So be it, until victory is mine and I have finished my shit — fuck yeah and amen.
* * *
The song is called "The Mirror of my Thoughts" and it's about loving a person from a photograph. It uses poetry as lyrics and it's very sentimental. Try listening to it while reading along.
(Sorry I couldn't translate it better... I'm kinda in the mood for karaoke now! My next pick will be something more rock-y.)
Seduta sulla riva di questa mia coscienza
Sitting in the shore of my conscience
la malinconia,
the melancoly,
far conto dei miei giorni
keeping account of my days
Comprese le esperienze di questa vita mia
including the experiences of this life of mine
Somiglia ad un gabbiano con le ali aperte lungo l'orizzonte
it looks like a seagull with its wings spread along the horizon
Abbraccia il cielo che è sopra di me
it hugs the sky that is above me
che affonda dentro lo scenario di un tramonto
which dives into the scenery of a sunset.
( LyricsCollapse )
My interpretation: It seems to be from the perspective of a whore who reminds herself bitterly to stay level-headed and pragmatic. The singer could be drawing from her biography, as she used to work for a "show club" before being discovered by the discographic company.
But I think any woman can identify on the part about "fighting daily battles"... Heck, I did when I was cleaning the bedsheets! >__>; It gets me every time. She's saying 'I'm not a princess, I have to work hard', and she shouts it too. Go girl!
The rush in the heart
e' già una novità
is already a novelty
which behind a newspaper is
changing opinions.
And the evil of the day
is a few kilometres to the south
of my return
of my goodmorning.
To the wacky music players
To the ludicrous like me
To those who never liked me
To those I've never met
for which I wonder why.
Ai dimenticati,
To the forgotten,
to the ended playboys,
and to me as well.
BUT the next day I found a stubbornly positive teacher's blog which saved me. I regained my will to teach and got motivated, even excited about it. I'm actually looking forward to classes now. I was able to remember why I enjoy teaching. So, THANK YOU, Christina C!
EDIT: It's getting better and better. I got them motivated to speak in English without slipping into their mother tongue, I made them rejoice in their success and feel they've achieved a step, and I've organised everything. The method is working~ OH YEAH! =)


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29 Mar 2010
Madama Butterfly, NYCO
Once again, as in L'Etoile, Mark Lamos's staging and Robert Wierzel's lighting nearly steal the show in the City Opera's revival of Madama Butterfly.
Penny Woolcock's 2010 production of Bizet's The Pearl Fishers returned to English National Opera (ENO) for its second revival on 19 October 2018. Designed by Dick Bird (sets) and Kevin Pollard (costumes) the production remains as spectacular as ever, and ENO fielded a promising young cast with Claudia Boyle as Leila, Robert McPherson as Nadir and Jacques Imbrailo as Zurga, plus James Creswell as Nourabad, conducted by Roland Böer.
Francesco Cavalli's La Calisto was the composer's fifteenth opera, and the ninth to a libretto by Giovanni Faustini (1615-1651). First performed at the Teatro Sant'Apollinaire in Venice on 28th November 1651, the opera by might have been sub-titled 'Gods Behaving Badly', so debauched are the deities' dalliances and deviations, so egotistical their deceptions.
At the end of Shakespeare's A Midsummer Night's Dream, Theseus delivers a speech which returns to the play's central themes: illusion, art and the creative imagination. The sceptical king dismisses 'The poet's vision - his 'eye, in a fine frenzy rolling' - which 'gives to airy nothing/ A local habitation and a name'; such art, and theatre, is a psychological deception brought about by an excessive, uncontrolled imagination.
Following the success of previous 'mini-festivals' at St John's Smith Square devoted to Schubert and Schumann, last weekend pianist Anna Tilbrook curated a three-day exploration of the work of Ralph Vaughan Williams and his contemporaries. The music performed in these six concerts was chosen to reflect the changing contexts in which it was composed and to reveal the vast changes in society, politics and culture which occurred during Vaughan Williams' long life-time (1872-1958) and which shaped his life and creative output.
Trying to work around Manon Lescaut's episodic structure, this new production presents the plot as the dying protagonist's feverish hallucinations. The result is a frosty retelling of what is arguably Puccini's most hot-blooded opera. Musically, the performance also left much to be desired.
It is Herodotus who tells us that when Xerxes was marching through Asia to invade Greece, he passed through the town of Kallatebos and saw by the roadside a magnificent plane-tree which, struck by its great beauty, he adorned with golden ornaments, and ordered that a man should remain beside the tree as its eternal guardian.
Poor Puccini. He is far too often treated as a 'box-office hit' by our 'major' opera houses, at least in Anglophone countries. For so consummate a musical dramatist, that is something beyond a pity. Here in London, one is far better advised to go to Holland Park for interesting, intelligent productions, although ENO's offerings have often had something to be said for them.
With only four singers and a short-story-like plot Don Pasquale is an ideal chamber opera. That chamber just now was the 3200 seat War Memorial Opera House where this not always charming opera buffa is an infrequent visitor (post WWII twice in the 1980's after twice in the 40's).
"Yang sementara tak akan menahan bintang hilang di bimasakti; Yang bergetar akan terhapus." ("The transient cannot hold on to stars lost in the Milky Way; that which quivers will be erased.") As soprano Tony Arnold sang these words of Tony Prabowo's chamber opera Pastoral, with astonishingly crisp Indonesian diction, the first night of the second annual Momenta Festival approached its end.
Some operas seemed designed and destined to raise questions and debates - sometimes unanswerable and irresolvable, and often contentious. Termed a dramma giocoso, Mozart's Don Giovanni has, historically, trodden a movable line between seria and buffa.
Péter Eötvös' The Sirens Cycle received its world premiere at the Wigmore Hall, London, on Saturday night with Piia Komsi and the Calder Quartet. An exceptionally interesting new work, which even on first hearing intrigues: imagine studying the score! For The Sirens Cycle is elegantly structured, so intricate and so complex that it will no doubt reveal even greater riches the more familiar it becomes. It works so well because it combines the breadth of vision of an opera, yet is as concise as a chamber miniature. It's exquisite, and could take its place as one of Eötvös's finest works.
New from Oehms Classics, Walter Braunfels Orchestral Songs Vol 1. Luxury singers - Valentina Farcas, Klaus Florian Vogt and Michael Volle, with the Staatskapelle Weimar, conducted by Hansjörg Albrecht.
Manitoba Underground Opera took audiences on a journey — literally and figuratively — as it presented its latest installment of repertory opera between August 19–26.
On a recent weekend Lyric Opera of Chicago gave its annual concert at Millennium Park during which the coming season and its performers are variously showcased. Several of the performers, who were featured at this "Stars of Lyric Opera" event, are scheduled to make their debuts in Lyric Opera's new production of Wagner's Das Rheingold beginning on 1 October.
Desire and deception; Amor and artifice. In Jan Philipp Gloger's new production of Così van tutte at the Royal Opera House, the artifice is of the theatrical, rather than the human, kind. And, an opera whose charm surely lies in its characters' amiable artfulness seems more concerned to underline the depressing reality of our own deluded faith in human fidelity and integrity.
On September 22, 2016, Los Angeles Opera presented Darko Tresnjak's production of Giuseppe Verdi's opera Macbeth. Verdi and Francesco Maria Piave based their opera on Shakespeare's play of the same name.
On September 18th, at a casual Sunday matinee, Pacific Opera Project presented a surprising choice for a small company. It was Igor Stravinsky's 1951 three act opera, The Rake's Progress. It's a piece made for today's supertitles with its exquisitely worded libretto by W.H. Auden and Chester Kallman.
We are nearing the end of Classical Opera's MOZART 250 sojourn through 1766, a year that the company's artistic director Ian Page admits was 'on face value a relatively fallow year'. I'm not so sure: Jommelli's Il Vogoleso, performed at the Cadogan Hall in April, was a gem. But, then, I did find the repertoire that Classical Opera offered at the Wigmore Hall in January, 'worthy rather than truly engaging' (review). And, this programme of Haydn and his Czech contemporary Josef Mysliveček was stylishly executed but did not absolutely convince.
Globalization finds its way ever more to San Francisco Opera where Italian composer Marco Tutino's La Ciociara saw the light of day in 2015 and now, 2016, Chinese composer Bright Sheng's Dream of the Red Chamber has been created.
Renowned Polish tenor Piotr Beczala and well-known collaborative pianist Martin Katz opened the San Diego Opera 2016–2017 season with a recital at the Balboa Theater on Saturday, September 17th.
Once again, as in L'Etoile, Mark Lamos's staging and Robert Wierzel's lighting nearly steal the show in the City Opera's revival of Madama Butterfly.
It says a great deal in her favor that in her sensitive and passionate account of the title role, Shu-Ying Li holds her own against such competition. For example, a great circle dominates the backdrop — red against white for the rising sun or the Japanese flag as the opera opens, turning black against red when the Bonze denounces his errant niece for renouncing her ancestral gods, then white against serene blue for moonlight during the great duet that closes the act.
Quinn Kelsey as Sharpless, Eddie Schweighardt as Sorrow and Shu-Ying Li as Cio-Cio San
There are few props on the wide, bare stage, and therefore those we see possess greater significance. The tiny ancestral gods that Pinkerton scorns as "dolls" are placed stage center, as on an altar, during later acts of the story — first for Butterfly to ignore them, preferring the wide-awake American god, then for her to atone before them by hara-kiri when she has lost honor, child and everything else. A fleet of tiny red "warships" floats like a flock of birds above the stage at curtain-rise — to be echoed in Act II by the toy ship Sorrow plays with. A telescope is not merely functional, permitting Butterfly to stare at the empty harbor of Nagasaki, but also symbolizes her undying hopes.
The traditional screen behind which the heroine kills herself does not appear — instead she shows her gumption (and forfeits a pathos playwright David Belasco and composer Puccini both wished to claim) by turning about to slash her throat in her faithless lover's face, precisely as Pinkerton enters and after his desperate, perhaps repentant cries of "Butterfly!" This confrontational death is an excess. Butterfly, as Belasco and Puccini conceived her, may be a convenient myth, a construction of the guilty Western colonialist conscience, but that is how they created her; to turn her final act into an expression of vendetta, a spitting in the face of her oppressor — the man she has entirely loved hitherto — seems an intrusive modern afterthought. We may have been patronizing when we took Cio-Cio-San to our hearts — but this resentful figure, this image of hate, will not win our hearts at all.
Shu-Ying Li as Cio-Cio San and Steven Harrison as B.F. Pinkerton
Shu-Ying Li has a large and genuine Puccini soprano of great beauty, but there is a certain unsteadiness until it warms up. Her entrance was not luminous, but the duet was, and many other soaring phrases as well. Her Cio-Cio-San was a complete portrait, each line part of the whole, not the collection of pretty excerpts it can become. She is a tall woman and does not move like a small one, and in her enormous wedding kimono she all but dwarfs her Pinkerton, but the robe is soon removed — it is present, hanging on rods, for the rest of the opera, a visible symbol of her past — and in slimmer robes she manages to appear becomingly frail.
Jessica Klein as Kate Pinkerton, Nina Yoshida Nelsen as Suzuki, Quinn Kelsey as Sharpless, and Shu-Ying Li as Cio-Cio San
Broad-shouldered Steven Harrison was, like most modern Pinkertons, cast in the shade by his Butterfly. The role may be thankless, an exemplary cad, but he should be a robust cad and Harrison sounded provincial. Nina Yoshida Nelsen sang an effective Suzuki, especially impressive in the Flower Duet and the penultimate trio. Quinn Kelsey's Sharpless possessed — not only because of his size — a real presence, so that one accepted the judgments of this involved, sympathetic observer as more than the official boilerplate they can seem. Little Eddie Schweighardt made one of the liveliest Troubles I've seen, whacking Jeffrey Halili's Goro with a fan and chasing about the stage with his toy boat.
George Manahan drew a precise and lustrous account of the score from his orchestra, full of Italian Wagnerism in the way melodies were foreshadowed, then foregrounded, then fragrantly recollected to remind us of joys past or hint at doom to come.
John Yohalem


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text:
Which Joe gave his name to 'sloppy joes'? We look at five interesting sandwiches and their lexical origins.
A piece of rock or metal that has fallen to the earth's surface from outer space as a meteor. Over 90 per cent of meteorites are of rock while the remainder consist wholly or partly of iron and nickel.
- 'The Martian meteorite crashes through the Earth's atmosphere - a blazing fireball.'
- 'A good case has already been made for bacteria microfossils in a Martian meteorite.'
- 'It is brought to us by such solar system visitors as meteorites, which burn up or fragment and drift as fine particles through the atmosphere.'
- 'Other sources of high iridium concentrations are extraterrestrial objects, such as meteorites or comets.'
- 'That is how fragments of the asteroid belt can end up crashing into Earth as meteorites.'
- 'It is possible that something may have been at the end of this, a piece of rock, a meteorite.'
- 'The age of the carbonates can be clearly determined, and some very old ones had been found on certain asteroids and meteorites.'
- 'It has been identified in carbonaceous achondrite and iron meteorites.'
- 'One possible source of such untainted data are Earth's lunar meteorites.'
- 'When meteorites hit nearby worlds, they kick up bits of rock, some of which might have enough speed to escape from their planet entirely.'
- 'A safe containing moon rocks and meteorites was stolen from the Johnson Space Center in Houston on July 15.'
- 'On display are minerals, fossils, meteorites, and some rocks, most of which are of reference quality.'
- 'Other Moon rocks brought back by the Apollo astronauts have helped in understanding the lunar meteorites found on Earth.'
- 'They are very similar in composition to the carbonaceous chondrite meteorites that sometimes fall on Earth.'
- 'Small impactors deliver meteorites, while large ones infrequently wreak global devastation.'
- 'As the newly launched Sputnik satellite orbits earth, a strange meteorite approaches the planet.'
- 'About 1.8 billion years ago, a meteorite or comet the size of Mount Everest slammed into what is now Canada.'
- 'Grouped with iron meteorites, these are pieces from the cores of fledgling planets destroyed by collisions when the Solar System formed.'
- 'Meteorites contain platinum and meteorites have been landing on earth for billions of years already.'
- 'The most recent major extinction occurred 65 million years ago when a meteorite crashed into Earth, leading to the demise of the dinosaurs.'
Further reading
12 synonyms for foolRead more
Are you looking for a word for a foolish person? We explore twelve interesting words to describe the dunderheads in your life.
6 'run' phrases you probably don't knowRead more
Before you run for the hills, let's run through a list of 'run' expressions that are running through our minds.
How brothers became buddies and brosRead more
The definitions of 'buddy' and 'bro' in the OED have recently been revised. We explore their history and increase in popularity.