Since patients with NAFLD have altered intestinal microbiota (IM), there is the potential for altered signaling of the chief TLR4 ligand, gram-negative endotoxin and other IM derived TLR ligands. Aims: 1. To compare hepatic expression of genes involved in TLR signaling between patients with simple steatosis (SS) or nonalcoholic steatohepatitis (NASH) Selleck Smoothened Agonist and healthy controls (HC); 2. To determine whether these genes correlate with IM. Methods:

In a cross-sectional study, gene expression (Illumina Whole-Genome DASL HT-assay) was measured in liver tissue obtained from biopsies of 20 patients with SS and 19 with NASH or during live donor hepatectomy (HC: n=24). In a subgroup (6 SS, 9 NASH, 8 HC), the relative

amounts of fecal Bifidobacteria, E. coli, Bacteroides/Prevotella, and Firmicutes selleckchem were assessed (qPCR). ANOVA with Tukey’s HSD test, Wilcoxon test, and Spearman correlations were applied. Results: Body mass index (BMI) and insulin resistance increased significantly from HC to SS and NASH. Bacteroides/Prevotella were lower in NASH compared to HC [median (IQR)] [0.90 (1.66) vs.3.45 (3.84) % of total bacteria; p=0.012) but not different from SS [3.03 (4.97) %]. Seven genes involved in TLR signaling were differentially expressed (>2.0-fold change, adjusted p<0.05) between NAFLD patients and HC. There were no differences between SS and NASH. Bacteroides/Prevotella (%) were correlated with TLR3, TLR7, and PELI1. When controlling for BMI, this remained significant: TLR3 (r=-0.85, p<0.0001), TLR7 (r=-0.599, p=0.0041), PELI1 (r=0.59, p=0.021). Conclusion: TLR signaling related genes are differentially expressed in NAFLD vs. HC, but expression profiles are similar in SS and NASH. The proportion of Gram-negative Bacteroides/Prevotella in stool was higher in NASH vs. HC and was correlated with the expression of important MCE公司 regulators of TLR signaling. The role of IM in TLR-induced inflammation and insulin resistance in NAFLD warrants further

study. Disclosures: Scott Fung – Advisory Committees or Review Panels: Merck, Vertex; Grant/Research Support: Gilead Sciences, Roche; Speaking and Teaching: Gilead Sciences, BMS The following Comelli, Marialena Mouzaki, Ian McGilvray, Sandra E. Fischer, Johane P. Allard Objective To evaluate cardiovascular benefits of modest alcohol consumption among men with nonalcoholic fatty liver disease (NAFLD). Design Cross-sectional study of 10, 581 consecutive male participants aged 40-69 years undergoing abdominal ultrasonography and carotid artery ultrasonography at the Center for Health Promotion of the Samsung Medical Center in Korea from January 2009 to December 2009. Results There were total 2, 129 men diagnosed with NAFLD, and the mean age of the participants was 52.6 years old.

Since patients with NAFLD have altered intestinal microbiota (IM), there is the potential for altered signaling of the chief TLR4 ligand, gram-negative endotoxin and other IM derived TLR ligands. Aims: 1. To compare hepatic expression of genes involved in TLR signaling between patients with simple steatosis (SS) or nonalcoholic steatohepatitis (NASH) this website and healthy controls (HC); 2. To determine whether these genes correlate with IM. Methods:

In a cross-sectional study, gene expression (Illumina Whole-Genome DASL HT-assay) was measured in liver tissue obtained from biopsies of 20 patients with SS and 19 with NASH or during live donor hepatectomy (HC: n=24). In a subgroup (6 SS, 9 NASH, 8 HC), the relative

amounts of fecal Bifidobacteria, E. coli, Bacteroides/Prevotella, and Firmicutes Cisplatin cost were assessed (qPCR). ANOVA with Tukey’s HSD test, Wilcoxon test, and Spearman correlations were applied. Results: Body mass index (BMI) and insulin resistance increased significantly from HC to SS and NASH. Bacteroides/Prevotella were lower in NASH compared to HC [median (IQR)] [0.90 (1.66) vs.3.45 (3.84) % of total bacteria; p=0.012) but not different from SS [3.03 (4.97) %]. Seven genes involved in TLR signaling were differentially expressed (>2.0-fold change, adjusted p<0.05) between NAFLD patients and HC. There were no differences between SS and NASH. Bacteroides/Prevotella (%) were correlated with TLR3, TLR7, and PELI1. When controlling for BMI, this remained significant: TLR3 (r=-0.85, p<0.0001), TLR7 (r=-0.599, p=0.0041), PELI1 (r=0.59, p=0.021). Conclusion: TLR signaling related genes are differentially expressed in NAFLD vs. HC, but expression profiles are similar in SS and NASH. The proportion of Gram-negative Bacteroides/Prevotella in stool was higher in NASH vs. HC and was correlated with the expression of important medchemexpress regulators of TLR signaling. The role of IM in TLR-induced inflammation and insulin resistance in NAFLD warrants further

study. Disclosures: Scott Fung – Advisory Committees or Review Panels: Merck, Vertex; Grant/Research Support: Gilead Sciences, Roche; Speaking and Teaching: Gilead Sciences, BMS The following Comelli, Marialena Mouzaki, Ian McGilvray, Sandra E. Fischer, Johane P. Allard Objective To evaluate cardiovascular benefits of modest alcohol consumption among men with nonalcoholic fatty liver disease (NAFLD). Design Cross-sectional study of 10, 581 consecutive male participants aged 40-69 years undergoing abdominal ultrasonography and carotid artery ultrasonography at the Center for Health Promotion of the Samsung Medical Center in Korea from January 2009 to December 2009. Results There were total 2, 129 men diagnosed with NAFLD, and the mean age of the participants was 52.6 years old.

009) and serum sodium (131 ± 7 versus 135 ± 5 mEq/L, P = 0.007) and higher blood urea nitrogen (32 ± 24 versus 24 ± 15 mg/dl, P = 0.06), plasma

renin activity (7.1 ± 9.9 versus 3.4 ± 5.6 ng/mL*h, P = 0.03), and noradrenaline concentration (544 ± 334 versus 402 ± 316 pg/mL, P = 0.02). During follow-up, patients with RAI exhibited a higher probability of infection (41% versus 21%, P = 0.008), severe sepsis (27% versus 9%, P = 0.003), type-1 hepatorenal syndrome (HRS) (16% versus 3%, P = 0.002), and death (22% versus 7%, P = 0.01). Conclusion: RAI is frequent in noncritically ill patients with acute decompensation of cirrhosis. As compared with those with normal adrenal function, patients with RAI have greater impairment of circulatory and renal function, higher probability of severe sepsis and type-1 HRS, and higher short-term mortality. (Hepatology 2013;58:1757–1765) Relative adrenal insufficiency (RAI) ABT-199 is a syndrome characterized by an inadequate production of cortisol with respect to peripheral demands.[1, 2] It was first described and has been mainly studied in critically ill patients (severe sepsis, septic shock, head injury, pancreatitis,

burns, and major surgery) who require high circulating cortisol levels to modulate systemic inflammatory response, maintain the vascular tone and permeability, and adapt metabolism to stress.[1-7] During the last decade evidence has been presented that RAI may also be an important feature in patients with cirrhosis.[8-22]

The first studies on RAI in critically ill patients with cirrhosis were published NVP-LDE225 cell line between 2003 and 2008 and included mainly patients with decompensated cirrhosis and severe sepsis or septic shock. These studies showed a very high prevalence of RAI (51%-77%) and a clear association with poor liver function, renal failure, medchemexpress refractory shock, and hospital mortality.[8-11] Two recent studies confirm the high prevalence of RAI in patients with cirrhosis and septic shock (76%)[12] or gastrointestinal hemorrhage (60%).[13] Recent but limited data suggest that RAI can also occur in noncritically ill cirrhosis patients.[14-22] The reported prevalence of this entity, which is now refer to as “hepatoadrenal syndrome,”[9, 22] ranges in the different studies between 7% and 49%, depending on the methodology used for RAI diagnosis.[15-22] However, there are no data on the relationship between RAI and clinical course of noncritically ill cirrhosis patients. This article reports the results of a prospective study evaluating adrenal function in a large series of patients admitted to the hospital for the treatment of acute decompensation of cirrhosis. Patients were followed for 3 months. The aim of the study was to assess if RAI is associated with differences in the natural course of the disease and survival.

2F,G). These data collectively demonstrate that epigenetic repression of the Pparγ gene in culture-activated HSCs is lifted by the

YGW extract treatment, and this effect must be responsible for restored PPARγ expression and HSC quiescence. Another important biochemical feature of activated HSCs is increased activity of nuclear factor kappaB FK506 purchase (NF-κB).24 We tested how the YGW extract affects this parameter. The treatment with the YGW extract markedly inhibits the activity of IκB kinase (IKK) as assessed by phosphorylation of IκBα-GST fusion protein (Fig. 3A), the expression of IκBα and β, both targets of NF-κB (Fig. 3B) in day-5 HSCs, and NF-κB promoter activity in the rat HSC line (BSC) (Fig. 3C). The demonstrated suppressive effects of YGW on IKK and NF-κB suggest

that it may promote apoptotic death of HSCs. Only after a prolonged extract treatment exceeding 4-5 days with replenishment of the medium containing the extract every 2 days does apoptosis of cultured HSCs begin to appear and become apparent after 8 days as assessed by TUNEL staining (Supporting Fig. 1A). As the first step in identifying active ingredients of YGW rendering the above reversal effects on activated HSCs, we first tested different fractions of gel filtration of the YGW water extract in culture-activated HSCs. This analysis revealed a fraction with a molecular mass range of 200 to 750 Da, reproduced the YGW effects including the morphological reversal (Fig. 4A), down-regulation of α1(I)procollagen mRNA (Fig. 4B), and decreased MeCP2 enrichment at the Pparγ promoter (Fig. 4C). This gel filtration fraction was next applied to LC/MS for identification of active Acalabrutinib clinical trial ingredients. This analysis identified small peaks with a retention time of 14 to 15 minutes (boxed in the UV254 tracing of Fig. 4D). Due to low amounts of these molecules detected in the water extract to allow their purification and identification, we next analyzed YGW ingredients extracted with butanol (BuOH). This method ensures that most hydrophilic and lipophilic organic compounds

are extracted into the butanol layer, while most of the sugar and ionic inorganic components remain in the water layer. After lyophilization, the water-soluble portion of YGW shows reduced activity of the HSC morphologic reversal when compared with the YGW water extract before butanol medchemexpress partitioning. In contrast, the butanol-soluble portion of YGW shows clear bioactivity toward HSCs (data not shown), suggesting that the bioactive phytocompounds are enriched in the butanol-soluble portion. We further fractionated the butanol-soluble portion by reverse phase chromatography eluted with 10% (A fraction), 40% (B fraction), and 100% (C fraction) acetonitrile-water mixtures (Fig. 4D). The butanol A fraction shows a reproducible effect on HSC morphologic reversal (Fig. 4E), whereas the C fraction causes immediate cytotoxicity evident by detachment of the cells (data not shown).

Non-assortative or mismatched pairs that had formed when females were first receptive could be gradually replaced by pairs matched for size through two mechanisms. First, males that are already paired can hold and assess two females simultaneously and tend to retain the female of higher quality in terms of fecundity and proximity to the moult (Dick, 1992). In this case, the shorter the female time left to the moult, the more the pairs previously made between mismatched pairs are likely to be broken because a large paired male would preferentially leave a small female for a bigger one (Dick & Elwood, 1989; Dick,

1992; Murata selleck screening library & Wada, 2002; Wada et al., 2011). Second, males can struggle for the access to a precopula female. Takeovers, where an unpaired male outcompetes a paired male in the access to a receptive female may account for positive size-assortative pairing because of the advantage of large males in both making

takeovers and resisting takeover attempts (Ward, 1983; Dick & Elwood, 1990). The ‘takeover hypothesis’ would predict that paired males would on average be larger in size when females are in late moulting PD0325901 order stages. However, our data do not support this hypothesis. Takeovers usually occur at a low frequency (Dick & Elwood, 1990) and their effect might be negligible in G. pulex (Franceschi et al., 2010). There is increasing evidence that the generation of size-assortative pairing is not entirely male determined and that female resistance also plays a role in the pairing process (Jormalainen, 2007; Wellborn & Cothran, 2007). Female resistance to pairing attempts are interpreted either as a form of female choice or as a way to shorten precopula duration to reduce the costs

associated to guarding (Jormalainen, Merilaita & Riihimaki, 2001; Cothran, 2008). Female resistance could become more important through time, so that the more mismatched the initial pairings, the more rapidly the female may be able to dislodge the male (Jormalainen, 1998; Sutherland, Hogg & Waas, 2007). Only larger males may be able to keep hold on or subdue resisting females (Elwood et al., 1987; Jormalainen, Merilaita & Hardling, 2000; MCE公司 Sutherland et al., 2007). Consequently, under the scenario of female resistance, the size of females found paired should differ according to moult stage, with a pattern where only males hold small females and/or large males hold large females in the early premoult stages. We did not find any difference (or any tendency) in mean female size between the premoult stages. This pattern suggests that G. pulex females are at least indifferent to males. If the costs associated to precopula paid by females are weak, female resistance is expected to be low or negligible. Accordingly, in G. pulex, early- and long-lasting precopula have been shown to confer on females some benefits by decreasing the intermoult duration (Galipaud et al., 2011). As such, pairs are likely to be formed early in the moult cycle.

[1, 9, 10] A minority of patients with idiopathic gastroparesis (19% in the National Institute of Diabetes and Digestive and Kidney Diseases Gastroparesis Clinical

Research Consortium Registry study noted earlier[12]) report an initial infectious prodrome such as gastroenteritis or respiratory infection. It has been suggested that idiopathic gastroparesis of acute onset with infectious prodrome could constitute postviral or viral injury to the neural innervation of the stomach or the interstitial cells of Cajal in the stomach. Differential diagnosis of gastroparesis entails excluding other possible causes including gastric SCH772984 outlet obstruction, peptic ulcer disease, neoplasm, small bowel obstruction, and inflammatory bowel disorder.[9] For evaluating gastric motor function, the standard test is gastric emptying scintigraphy, which uses a labeled isotope bound to solid food to image gastric motility.[9, 13] Examples of normal and delayed gastric emptying rates measured with gastric emptying scintigraphy after consumption of a low-fat egg-white sandwich meal with water are shown in the Figure. Use of a wireless motility capsule to quantify luminal pH

Akt inhibitor and pressure is an alternative to gastric emptying scintigraphy.[9] This test measures whole-gut transit – that is, gastric emptying, small bowel transit, and colonic transit. Gastric emptying is manifested by a significant, sustained increase in pH as the capsule passes from the acidic stomach to the alkaline small intestine. Breath tests, another alternative to gastric emptying scintigraphy, measure radiolabeled CO2 in exhaled breath samples after duodenal processing of a consumed substrate.[9] Findings generally correlate well with results of gastric emptying scintigraphy. This test is used clinically in Europe, whereas in the United States, breath tests are most often employed in research studies and rarely

used in the clinic. While gastric motor testing is useful 上海皓元 in diagnosing gastroparesis, it has drawbacks. First, gastric emptying rates measured by gastric motor testing generally correlate poorly with symptoms and quality-of-life impact of gastroparesis.[14, 15] This finding suggests that factors in addition to slow gastric emptying contribute to symptoms. Relatively high interindividual and intraindividual variability in gastric emptying rates measured with gastric motor testing constitutes another limitation of gastric motor testing.[9] The relative contributions to these variabilities of gastric motor testing methodology and biologic inconsistency in gastric emptying are not currently known. Management of gastroparesis is guided by the goals of correcting fluid, electrolyte, and nutritional deficiencies; identifying and treating the cause of delayed gastric emptying (eg, diabetes); and suppressing or eliminating symptoms.

[1, 9, 10] A minority of patients with idiopathic gastroparesis (19% in the National Institute of Diabetes and Digestive and Kidney Diseases Gastroparesis Clinical

Research Consortium Registry study noted earlier[12]) report an initial infectious prodrome such as gastroenteritis or respiratory infection. It has been suggested that idiopathic gastroparesis of acute onset with infectious prodrome could constitute postviral or viral injury to the neural innervation of the stomach or the interstitial cells of Cajal in the stomach. Differential diagnosis of gastroparesis entails excluding other possible causes including gastric 5-Fluoracil chemical structure outlet obstruction, peptic ulcer disease, neoplasm, small bowel obstruction, and inflammatory bowel disorder.[9] For evaluating gastric motor function, the standard test is gastric emptying scintigraphy, which uses a labeled isotope bound to solid food to image gastric motility.[9, 13] Examples of normal and delayed gastric emptying rates measured with gastric emptying scintigraphy after consumption of a low-fat egg-white sandwich meal with water are shown in the Figure. Use of a wireless motility capsule to quantify luminal pH

PLX4032 cost and pressure is an alternative to gastric emptying scintigraphy.[9] This test measures whole-gut transit – that is, gastric emptying, small bowel transit, and colonic transit. Gastric emptying is manifested by a significant, sustained increase in pH as the capsule passes from the acidic stomach to the alkaline small intestine. Breath tests, another alternative to gastric emptying scintigraphy, measure radiolabeled CO2 in exhaled breath samples after duodenal processing of a consumed substrate.[9] Findings generally correlate well with results of gastric emptying scintigraphy. This test is used clinically in Europe, whereas in the United States, breath tests are most often employed in research studies and rarely

used in the clinic. While gastric motor testing is useful MCE in diagnosing gastroparesis, it has drawbacks. First, gastric emptying rates measured by gastric motor testing generally correlate poorly with symptoms and quality-of-life impact of gastroparesis.[14, 15] This finding suggests that factors in addition to slow gastric emptying contribute to symptoms. Relatively high interindividual and intraindividual variability in gastric emptying rates measured with gastric motor testing constitutes another limitation of gastric motor testing.[9] The relative contributions to these variabilities of gastric motor testing methodology and biologic inconsistency in gastric emptying are not currently known. Management of gastroparesis is guided by the goals of correcting fluid, electrolyte, and nutritional deficiencies; identifying and treating the cause of delayed gastric emptying (eg, diabetes); and suppressing or eliminating symptoms.

4A). Similar trends were observed from the samples treated with GW4064 for a short time only (1 hour) (Supporting Fig. 12), which is consistent with a direct effect of FXR on gene expression, rather than delayed indirect responses. For the genes unique to obese mice, mRNA levels were increased significantly for 5 of 13 genes and decreased significantly in one (Fig. 4B). These results suggest that a large fraction of potential FXR target genes examined are likely repressed by agonist-activated FXR. Direct gene repression by agonist-activated FXR was expected to be rare, because

Ibrutinib manufacturer nearly all of the direct FXR target genes have been reported to be activated.2, 3, 19 We therefore further examined epigenetic

histone markers of gene activation and repression, as well as occupancy by RNAPII.21, 24, 25 Genes with increased expression, as well as representative genes that were repressed or not Small molecule library concentration significantly affected (Fig. 4A), were examined. Two-step re-ChIP and qRT-PCR analyses were performed, and the known FXR target gene, Shp, was first analyzed as a control. Co-occupancy of RNAPII with FXR and acetylated histone H3K9/K14 at the Shp promoter was increased, whereas co-occupancy of RXRα with FXR was not changed after GW4064 treatment (Fig. 5A). As expected, mRNA levels for Shp were increased after a 1-hour treatment with GW4064 (Fig. 5B). For selected potential FXR genomic targets, occupancy of FXR was increased in nearly all of the genes examined after GW4064 treatment (Fig. 5C). For the three genes with increased mRNA levels after GW4064 treatment (Fig. 4A), aldehyde dehydrogenase 1 and 2 (Aldh1/2), 3-oxoacid coenzyme A transferase 2B (Oxct2b), and cell division cycle-associated 4 (Cdca4), co-occupancy of RXRα and RNAPII with FXR was increased and acetylated histone H3K9/K14 levels were increased, medchemexpress as expected, for activated genes (Fig. 5D-F). For the remaining genes, RNAPII

occupancy and acetylated histone H3 levels were decreased or unchanged, which would be consistent with either no induction or suppression of these genes, as observed from the mRNA levels. Finally, to directly determine whether binding of agonist-activated FXR leads to the repression of genes examined, ChIP assays were performed to measure the levels of known epigenetic histone marks for gene repression, such as H3K9 di- and H3K27 tri-methylation.24, 25 After treatment with GW4064, occupancies of FXR and RNAPII were increased and tri-methylated H3K9 and H3K27 levels were decreased at the control Shp gene promoter (Fig. 6A). For the potential FXR target genes with increased expression, such as Ald1/2, Oxc2b, and Cdca4, levels of repressed histone marks (H3K27 and H3K9) were unchanged or decreased after GW4064 treatment (Fig. 6B).

, 1996) with

a readily available, evenly distributed and relatively stable food source (Dostine & Franklin, 2002), all of which could presumably reduce extrinsic mortality. In our comprehensive multivariate analysis, breeding sociality significantly affected mean maximum longevities of avian families (Table 2; Appendix 3). A posteriori analyses revealed that social species lived longer than non-social species (Fig. 4). These results agree with those of Arnold & Owens (1998), who reported that cooperative breeding was correlated with low annual mortality and long life spans, as predicted by kin selection this website theory and life-history theory (Bourke, 2007). However, subsequent analyses by Møller (2006) and Blumstein & Møller (2008) called into question the role of sociality in the evolution of Linsitinib mw avian longevities and senescence patterns. The reasons for the difference between our results and theirs probably lie in differences

in both sample sizes and definitions of sociality. Whereas Møller (2006) defined sociality as ‘colonial nesting’ and Blumstein & Møller (2008) defined it as ‘cooperative breeding,’ our definition included both. We took the broader approach because both colonial nesting and cooperative breeding have often been linked to reduced predation rates on adult birds, chicks and eggs, due to shared vigilance, sentinels, alarm calling, cooperative group defense, safety in numbers and selfish herd effects (e.g. Hoogland & Sherman, 1976; Hoogland, 1981; Hailman, McGowan & Woolfenden, 1994; Clutton-Brock et al., 1999; Hatchwell

& Komdeur, 2000; reviewed by Safran et al., 2007). The link between sociality and longevity is illustrated by the characteristics of the four longest and shortest-lived avian orders 上海皓元医药股份有限公司 (Fig. 1). All four species of Phoenicopteriformes (flamingos) in our data base (Appendix 2) breed in colonies and crèche their chicks, all 25 Procellariiformes (petrels and shearwaters) and all 16 Pelecaniformes (pelicans) nest colonially, and 25 of 47 species (54%) of Psittaciformes (parrots) nest colonially or breed cooperatively. By contrast, only 38 of 179 (21%) Passeriformes (perching birds) and only two of nine (22%) Columbiformes (pigeons) in our data base nest colonially or breed cooperatively, only one of four (25%) Podicipediformes (grebes) breeds colonially, and only three of 15 (20%) Piciformes (woodpeckers) breed cooperatively.

19 This led us to evaluate the potential role of β2SP in mouse and human liver regeneration

and, specifically, the activation of hepatic progenitor cells. Our initial analysis reveals that β2SP expression demonstrates a clear spatial and temporal CHIR-99021 chemical structure variation as regeneration proceeds and has a reciprocal relationship with the expression of several progenitor cell markers. Reduced β2SP is also associated with an expanded population of hepatic progenitor cells following two-thirds partial hepatectomy that are likely activated by impaired hepatocyte proliferation and activated Wnt signaling in β2SP+/− mutant mice. β2SP, β2-Spectrin; DDC, 3,5-diethoxycarbonyl-1.4-dihydrocollidine; ES, embryonic stem; HCC, hepatocellular carcinoma; Oct3/4, octamer 3/4; PH, plekstrin homology; STAT3, signal transducer AZD2014 cost and activator of

transcription 3; TBRII, TGF-β type II receptor; TGF-β, transforming growth factor beta; TRITC, tetramethyl rhodamine isothiocyanate. Formalin-fixed and paraffin-embedded human postliving donor transplant liver biopsy specimens were obtained from the Department of Pathology, Georgetown University Medical Center, Washington, DC. Liver biopsies from 10 living donor transplant recipients were collected at 1 week (two specimens), 6 weeks (five specimens), and 12–16 weeks (three specimens) posttransplant as part of a standardized protocol to rule out liver pathology following living donor transplantation. Zero specimens were collected to evaluate for suspected rejection. All human tissue procedures were approved by the Institutional Review Board of Georgetown University Medical Center, Washington, 上海皓元 DC. Wild-type

and β2SP+/− 129 SvEv Black Swiss mice 8–16 weeks of age were subjected to two-thirds partial hepatectomy as described by Mitchell and Willenbring20 and then sacrificed at 0, 24, 48, 72, and 168 hours after hepatectomy (n ≥ 3). Liver tissue was then collected for immunohistochemical, protein, and RNA analysis. Generation of β2SP+/− knockout mice was as described.16 Whole-cell lysates were prepared from pooled livers from each experimental group with a radioimmunoprecipitation assay buffer (Sigma) containing fresh protease and phosphatase inhibitor cocktails. The primary antibodies used in this study were rabbit anti-β2SP (1:1000) and rabbit anti-actin (1:2500). Details of anti-β2SP antibody have been described.16 Horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnology) were used at 1:5000 dilution.