However, immunogenetic influence has been poorly investigated and

However, immunogenetic influence has been poorly investigated and mainly confined to HLA-class

II serological polymorphisms, because of their central role in the adaptive response. Nevertheless, it has been suggested that the role of the immune defense system, as well as the relevance of the genetic background, could better explain the pathogenesis of HCV infection, and these factors have been examined.10, 11 In adult patients, genetic variations in the IL28B gene, an innate cytokine, have been associated with the response to IFN-α/ribavirin therapy and spontaneous clearance in HCV genotype 1.26-28 For this reason, we evaluated the role of Quizartinib mw IL28B polymorphism in HCV genotype 1 vertical transmission, transient viremia, and chronic infection in infants. This is the first study that attempts to describe both HCV-VT and the spontaneous clearance of HCV, taking into account the influence Selleck Sotrastaurin of IL28B polymorphism

in mothers and children. The data obtained indicate that the IL28B genotype of mothers and children does not influence HCV-VT. Nevertheless, in the chronic infection study, 83% of the infants with the CC genotype exhibited spontaneous clearance (transient viremia) versus only 22% of the children with a non-CC genotype. On the other hand, the maternal IL28B genotype did not influence HCV chronic infection. Multivariate analysis identified the infant’s Rs12979860 CC IL28B genotype as the only factor independently associated with the spontaneous clearance Prostatic acid phosphatase of HCV. To the best of our knowledge, the present study is the first one to identify IL28B Rs12979860 polymorphism as a predictor of HCV spontaneous clearance in infants infected with HCV genotype 1 by vertical transmission. More information is now needed to understand the mechanisms that underlie this association, as well as the clinical impact of IL28B polymorphisms on HCV infection. The multivariate

analysis performed clearly shows the distinction between the risk factors in HCV-VT and in chronic infection. In HCV-VT, a high HCV viral load was independently associated with HCV-VT, thus confirming the bivariate analysis and the data previously published, by ourselves and by others. These data suggest that the maternal characteristics are more important in HCV-VT than are those of the infants. However, in the chronic HCV infection study, the multivariate analysis showed that the only factor independently associated with HCV clearance was the infants’ IL28B genotype, which confirmed our hypothesis that in infected infants the host’s immunogenic influence is crucial to the HCV viral response. Finally, all retrospective analyses have inherent limitations, but we have tried to minimize their effects.

Results: Gallbladder was enlarged in 35 patients (89 [86-106] mL)

Results: Gallbladder was enlarged in 35 patients (89 [86-106] mL) and within normal

range in 42 (41 [35-47] mL). Patients with enlarged gallbladders did not significantly differ from others regarding gender (65% vs. 66% males), median age (43 vs. 42 years), time from diagnosis (5 vs. 5.5 years), body mass index (21.6 vs. 23.7), associated inflammatory bowel disease (71% vs. 50%), UDCA treatment (89% vs. 90%), other MRI features including cystic abnormalities (8.5% vs. 12%), clinical or histological parameters of liver disease (Mayo risk score of −0.18 [−0.45-0.27] vs. −0.005 [−0.63-0.56]). Notably, malignancy was less frequent in the group with enlarged gallbladder, occurring in 2 (5.7%) vs. 11 (26.2%) patients with normal gallbladder size (P=0.029). Colorectal cancer in particular was 6.7-fold less frequent, occurring in 1 (2.8%) Akt inhibitor vs. 8 (19%) patients (P=0.037, OR=6.7 [0.9- 354])). In patients with enlarged gallbladder, the serum concentrations of secondary bile acids were lower than in other patients (1.6 [1.3-1.9] vs. 2.5 [2-3.1]

μmol/L, P=0.0004). This was true for deoxycholic acid (0.7 [0.5-1] vs. 2.2 [1-6-3] μmol/L, P=0.0001), a secondary bile acid known to promote colon carcinogenesis. Patients in this group also had higher concentrations of primary bile acids (10.5 [6.6-16.7] vs. 4.3 [3.5-5.3] μmol/L, P=0.0001) selleck chemicals and of UDCA (44.0 [29.4-52] vs. 27.2 [14.6-31.1] μmol/L, P=0.001). Furthermore, they had higher serum concentrations of the gallbladder-relaxing hormone FGF19 (211.6 [168.6-234.6] vs. 88.6 [72.7-121.6] pg/mL, P=0.0001), 4��8C which concentration was correlated with gallbladder volume (R2=0.46, P=0.001) Conclusion: Gallbladder is enlarged in approximately half of PSC patients, which can be caused by increased FGF19 levels, and which is associated with a lack of secondary bile acids, enhanced UDCA enrichment and a lower prevalence of colorectal cancer, consistent with protective properties of gallbladder enlargement in PSC. Disclosures: Olivier Chazouillères – Consulting: APTALIS, MAYOLY-SPINDLER The following

people have nothing to disclose: Mourad Aissou, Lionel Arrivé, Dominique Rainteau, Sara Lemoinne, Astrid Donald D. Kemgang Fankem, Delphine Firrincieli, Nicolas Chignard, Christophe Corpechot, Chantal Housset [Background and Aims] Although it is well established that treatment with ursodeoxycholic acid (UDCA) improves long-term outcome in patients with primary biliary cirrhosis (PBC), it is still uncertain whether “early” PBC, with normal or low ALP levels and at early histological stages, would benefit from UDCA treatment. In Japan nationwide surveys for PBC have been performed every three year since 1980, and so far clinical data of 7,376 cases have been accumulated. In the current study we examined the long-term outcome of asymptomatic PBC patients with normal or low ALP and at early histological stages in whom UDCA treatment was not initiated.

12 Accordingly, in our current study we confirmed that UDCA-LPE w

12 Accordingly, in our current study we confirmed that UDCA-LPE was able to dampen the susceptibility of the liver toward extrinsic apoptosis as well as the inflammatory response, with a

pronounced down-regulation of mediators such as MCP1, VCAM1, or TNF-α, which are responsible for leukocyte recruitment to the site of inflammation. MCP1 has also been described to be involved in the process of Alpelisib mouse chemotaxis and fibrogenic activation of stellate cells,28, 29 leading to TGF-β1 and extracellular matrix production. Currently, our preliminary observations indicate that UDCA-LPE may be capable of impairing fibrotic response due to the MCD diet (unpublished data). Thus, based on ongoing experimental studies, potentially beneficial effects of UDCA-LPE on hepatofibrogenesis should be addressed in the MG132 future. Lipotoxicity attributable to potent proinflammatory lipid intermediates has been implicated in deteriorating parenchymal damage during NAFLD. The phospholipase A2 (PLA2) cleavage product LPC, which plays a pivotal role in different inflammatory conditions,30-32 mediates hepatocellular apoptosis due

to palmitate-induced lipotoxicity.33 Furthermore, LPC levels were found to be elevated in livers of NAFLD patients.4 Earlier studies showed that LPC abundance in mitochondria is able to induce hepatocellular death34 caused by mitochondrial membrane depolarization.35 Our results proved that UDCA-LPE is capable of lowering increased LPC pools in dietary NAFLD, as previously demonstrated for acute liver injury in vitro and in vivo.12 Additionally, HFD mice treated with UDCA-LPE displayed reduced serum activity of PLA2 (unpublished observations), which

Thiamet G was previously reported to be necessary for lipid droplet biogenesis.36 Thus, inhibition of PLA2 may serve as a further mechanism for how the conjugate prevents hepatic lipid accumulation by way of inflammation inhibition. Further experimental studies are needed to prove this hypothesis. Excessive hepatic fat deposits may further serve as a prerequisite for subsequent liver injury due to lipid peroxidation. Enhanced ROS formation in NASH may oxidize unsaturated lipids to generate lipid peroxidation products.37 In our study, treatment with UDCA-LPE achieved a marked reduction in lipid hydroperoxides in mice fed an MCD diet, which has been previously shown to cause extensive increase in these cytotoxic lipid byproducts.38 Hepatic fat accumulation accompanied by changes in lipid metabolism is an essential pathophysiological feature of NAFLD. In accordance with earlier studies in humans,39, 40 HFD mice displayed up-regulation of de novo lipogenesis with enhanced expression of FASN and ACC1 as well as a moderate increase in SREBP1c, which is largely responsible for the regulation of enzymes involved in fatty acid synthesis.

0 Results: Results Among the 2849 HBV complete genome sequences,

0. Results: Results Among the 2849 HBV complete genome sequences, 217(8%) strains with Y(I/V) DD were identified. Of them, 120 had YIDD mutation and 97 had YVDD mutation. 1543 strains (54.2%) with PC-BCP mutation, seven mutation patterns of G1896A, G1899A, G1896A-G1899A, A1762T/G1764A, A1762T/G1764A-G1896A, A1762T/G1764A-G1899A, A1762T/G1764A-G1896A-G1899A were identified. Among the Y(I/V) DD strains, 165 (76%) strains had PC-BCP mutations. YV/IDD mutation with higher incidence of PC-BCP mutations were detected than without YMDD mutation (76% vs 24.0%, χ2 = 45.283,

P = 0.000), YIDD mutation with higher incidence of PC-BCP mutations than YVDD mutation (85% vs 64.9%, χ2 = 11.836, P = 0.001) and lamivudine (LAM) resistance of YI/VDD mutation with higher incidence of PC-BCP mutations than pre-existent YI/VDD (89.3% vs 58.9%, χ2 = 27.084, P = 0.000). The three patterns of G1896A-G1899A BTK inhibitor mutation (P = 0.000, OR = 7.573), A1762T/G1764A- G1899A mutation (P = 0.000, OR = 6.539) and A1762T/G1764A-G1896A-G1899A mutations (P = 0.000, OR = 6.596) have a higher tendency to develop YIDD mutation according to binary logistic analysis. Conclusion: Conclusion There is a relationship between HBV YI/VDD mutation and PC-BCP mutations. Different PC-BCP mutation patterns have different effect on YI/VDD mutation. Key Word(s): 1. Hepatitis B virus;

2. Y(I/V) DD mutation; 3. PC-BCP mutations; Table 1 Relative risks of YI/VDD according selleck inhibitor to Binary logisticl analysis for difference Pre Core-basic core

promotoe mutation pattens Factors YIDD YVDD 相对危险度 OR (95%CI) P 值 相对危险度 OR (95%CI) P 值 ? G1896A-G1899A 7.573 0.000 0 0.996 (3.77–15.212) (0–0) ? A1762T/G1764A 4.497 0.000 2.494 0.000 (2.729–7.410 ) (1.568–3.96) ? A1762T/G1764A+ G1896A 6.103 0.000 3.575 0.000 (3.474–10.725) (2.074–6.161) ? A1762T/G1764 A+ G1899A 6.539 0.000 0 0.997 (2.402–17.799) (0–0) ? A1762T/G1764A+G1896AA+1899G 6.596 0.000 0 0.997 (2.757–15.785) (0–0) Presenting Author: QIANQIAN ZHANG Additional Authors: XIAOLIN GUO Corresponding Author: QIANQIAN ZHANG, XIAOLIN GUO Affiliations: the first hospital of Jilin university Objective: Nucleoside Ureohydrolase analogues of antiretroviral drugs currently widely listed are entecavir, lamivudine, telbivudine, adefovir dipivoxil, each of which has its advantages and disadvantages. And entecavir with strong antiviral effect, rapid onset, low resistance, especially for patients with initial treatment or severe conditions, is the best choice for antiviral treatment at present. To observe the decrease of viral load of hepatitis B, the improvement of liver function and blood coagulation function of the patient of hepatitis B associated with acute on chronic liver failure after antiviral treatment with application of entecavir 1.0 mg/day, we reviewed 1 case with acute on chronic hepatitis B liver failure in our hospital. Methods: The patients of acute on chronic hepatitis B liver failure was administrated entecavir of 0.

Methods: The changes of intracellular Ca2+ ([Ca2+]i) in HCC cells

Methods: The changes of intracellular Ca2+ ([Ca2+]i) in HCC cells were examined using Alpelisib datasheet the Ca2+-sensitive dye Fura-2 AM. The cell proliferation was estimated with MTT and Edu assays, and the cell migration and invasion were estimated with scratch wound and transwell

assays. Results: TGF-β (10 ng/ml) rapidly stimulated [Ca2+]i increases in normal liver cell LO2 and hepatocellular carcinoma cell HepG2. Both 2-APB, a blocker of canonical transient receptor potential channels (TRPC6), and KB, a blocker of Na+/Ca2+ exchanger (NCX1) partially inhibited TGF-β-induced [Ca2+]i increase in HepG2(P < 0.05), and 2-APB had more markedly inhibitory effect than KB. In the absence of extracellular Ca2+, TGF-β did not induce significant the change of [Ca2+]i in HepG2 cells. The further study showed that the mRNA and protein expression levels of NCX1 and TRPC6 were obviously increased in HepG2 after incubation with TGF-β

for 24 hours. TGF-β promoted the cell proliferation, migration, and invasion in HepG2 cells, which was partially check details inhibited by both 2-APB and KB. Bapta-am, an intracellular calcium chelator, completely inhibited the effect of TGF-β on HepG2 cells. Conclusion: TGF-β regulates the cellular behavior of HCC through intracellular Ca2+ signal. TRPC6 and NCX1 were involved in the role of TGF-β on HCC cells. Key Word(s): 1. TGF-β; 2. HCC; 3. NCX1; 4. TRPC6; Presenting Author: BIGUANG TUO Additional Authors: RUI XIE, JINGYU XU, GUORONG WEN, HAI JIN, XUEMEI LIU, YUAN YANG, BEI JI, YIXIA JIANG, HUI DONG Corresponding Author: BIGUANG TUO Affiliations: Department of Gastroenterology, Affiliated Hospital of Zunyi Medical college Objective: P2Y2 receptor (P2Y2R) mediates a variety Thiamet G of biological functions. ATP is a physiologic ligand for P2Y2R, which can be released from inflammatory cells and tumor cells, and is an abundant biochemical component of the tumor microenvironment. It is well known

that chronic inflammation plays a key role in the development and progression of human hepatocellular carcinoma (HCC). In this study, we investigated the expression and role of P2Y2R in human HCC cells, which aimed to find a new therapeutic target against HCC. Methods: The experiments were performed in native isolated human HCC cells and normal hepatocytes, and human HCC cell lines. The xenograft model of human HCC was established in nude mice. Results: The mRNA and protein expressions of P2Y2R, not P2Y4R, in native human HCC cells and human HCC cell lines, HepG2 and BEL-7404, were markedly increased, compared to human normal hepatocytes and normal hepatocyte line LO2, respectively (P < 0.01 and P < 0.001). P2Y2R anatgonist suramin and specific siRNA, not P2Y4R specific siRNA, inhibited ATP-induced [Ca2+]i increases in HCC cells (P < 0.0001).

4 A total of 81 cases of patients with cirrhosis of the liver ba

4. A total of 81 cases of patients with cirrhosis of the liver basis, of which 39 patients received chemoembolization, the median survival was 4.08 months, 42 patients received embolization, the median survival of 3.5 months, The average of TTP, mediansurvival and survival curves difference between two groups of patients has not statistically significant.j Conclusion: TAE was effective and safety for primary hepatocellular carcinoma associated with ascites, leukopenia, and portal vein tumor thrombus, liver cirrhosis. Key Word(s): 1. TACE,; 2. HCC; 3. leukopenia; Presenting

Author: LILI DING Additional Authors: JUNPU GAO, QIJUN NIU Corresponding Author: LILI DING Affiliations: jilin university Objective: The objective of the present study was to explore Tamoxifen the different metabolic substances in sera between hepatocellular carcinoma (HCC) caused by hepatitis B virus (HBV) or hepatitis C virus (HCV). We used the metabonomic method to profiling the sera among liver cirrhosis, NVP-BKM120 liver cancer

and normal persons, and find potential tumor biomarkers. Methods: UPLC/Q-TOF-MS was utilized to profile the different metabolic substances in sera among 29 cases of liver cirrhosis which induced by HBV and HCV respectively, 38 and 32 cases of HCC sera which induced by HBV and HCV respectively, and

30 cases of normal persons. We got retention time, m/z and total electrories Verteporfin manufacturer strength related figure, then matched peaks, normalized these peaks, and analysed the data with PCA and PLS methods, matched these substances with human metabonomics database, the different substances between each team may be the potential biomarkers. Results: Metabolic substances in liver cirrhosis caused by HBV and HCV had no differences. Twenty-six substances were identified as potential tumor markers, ten compounds were identified as: LysoPC (20 : 1), LysoPC (20 : 2), LysoPC (P-18 : 0), LysoPC (P-18 : 0), LysoPC (15 : 0), arachidonic acid, hemolysis phosphatidyl choline, carnitine, glycine, tryptophan, N-Arachidonoyl glycine. Conclusion: There were no differences between liver cirrhosis between HBV and HCV induced liver cancer in metabolites level. Through analysising liver cirrhosis and liver cancer sera, Twenty-six different substances were found, LysoPC (20 : 1), LysoPC (20 : 2), LysoPC (P-18 : 0), LysoPC (P-18 : 0), LysoPC (15 : 0), arachidonic acid, hemolysis phosphatidyl choline, carnitine (C18 : 1), carnitine (C18 : 2), glycine, tryptophan, N-Arachidonoyl glycine, they are the potential tumor biomarkers of HCC. Key Word(s): 1. Metabonomics; 2. HCC; 3. liver cirrhosis; 4.

1) CD40L bisulfite sequencing data were obtained on a minimum of

1). CD40L bisulfite sequencing data were obtained on a minimum of seven clones prepared from each of CHIR-99021 cell line both CD4+ and CD8+ T cells isolated from the PBMCs of 20 PBC patients and 20 unrelated controls. The CD40L promoter sequences amplified from CD4+ T cells of PBC patients

showed significantly lower methylation, as compared to healthy controls. The methylation patterns of individually sequenced clones are shown for two representative subjects in Fig. 2A. Overall promoter methylation was determined as the percentage of methylated CpG sites of all possible CpG sites, which indicated a significant reduction in CD4+ T cells from patients, compared to healthy controls (0.54 versus 0.64; P < 0.001), to subjects with type I diabetes (0.54 in PBC versus 0.66; P < 0.001), and to psoriasis patients (0.54 in PBC versus 0.67; P < 0.001) (Fig. 2B). Similarly, site-specific methylation was calculated for each of the 10 CpG sites in the CD40L click here promoter region (Fig. 2C) and ranged from 0.33 to 0.61

in PBC patients versus 0.29-0.78 in healthy subjects, 0.29-0.78 in type I diabetes, and 0.32-0.75 in psoriasis patients (Fig. 2C). No detectable difference in the level of CD40L promoter methylation in isolated CD8+ T cells was observed between PBC patients and controls (data not shown). However, in general, the levels of CD40L promoter methylation were significantly downmethylated in CD4+ T cells, compared to CD8+ T cells from both PBC patients (Fig. 3A) and controls (data not shown), as confirmed by a significantly lower CD4+/CD8+ methylation ratio in PBC patients (0.72 versus 0.85; P = 0.0002) (Fig. 3B). Levels of CD40L mRNA were also evaluated in CD4+ T cells from both patients and healthy controls by reverse-transcriptase PCR. Levels of CD40L mRNA expression was increased in CD4+T cells from PBC patients, compared to controls (2−(ΔΔCt) = 2.53 versus 1.12; P = 0.0178) and inversely correlated with levels of CD40L promoter methylation (r2 = 0.2347, P = 0.0355; Fig. 4). Based on the fact that patients with

mutations of the X-linked CD40L gene exhibit high titers of serum IgM,18 we evaluated the potential correlation between levels of CD40L methylation and serum IgM levels. The sera from 16 of the 6-phosphogluconolactonase 20 PBC patients (80%) included in the study had high relative levels of IgM (Fig. 5A) and showed significantly lower levels of CD40L promoter methylation within CD4+ T cells, compared to their normal IgM counterparts (Fig. 5B). Interestingly, IgM levels inversely correlated with levels of CD40L promoter methylation (r2 = 0.5448, P = 0.0011; Fig. 5C). To determine the potential contribution of the presence of mutations of the CD40L gene that could influence IgM levels,17, 18 we sequenced CD40L in gDNA samples isolated from each of the PBC patients and analyzed them for the presence of mutations previously documented for the CD4L gene (Fig. 6).


“High levels of intraspecific variability are often associ


“High levels of intraspecific variability are often associated with HAB species, and this variability is likely an important factor in their competitive success. Heterosigma akashiwo (Hada) Hada ex Y. Hara et M. Chihara is an ichthyotoxic raphidophyte capable

of forming dense surface-water blooms in temperate coastal regions throughout the world. We isolated four strains of H. akashiwo from fish-killing northern Puget Sound blooms in 2006 and 2007. By assessing numerous aspects of biochemistry, physiology, and toxicity, we were able to describe distinct ecotypes VX-770 that may be related to isolation location, source population, or bloom timing. Contrasting elements among strains were cell size, maximum growth and photosynthesis rates, tolerance of low salinities, amino acid use, Selleckchem BMS-777607 and toxicity to the ciliate grazer Strombidinopsis acuminatum (Fauré-Fremiet). In addition, the rDNA sequences and chloroplast genome of each isolate were examined, and while all rDNA sequences were identical, the chloroplast genome identified differences among the strains that

tracked differences in ecotype. H. akashiwo strain 07A, which was isolated from an unusual spring bloom, had a significantly higher maximum potential photosynthesis rate (28.7 pg C · cell−1 · h−1) and consistently exhibited the highest growth rates. Strains 06A and 06B were not genetically distinct from one another and were able to grow

on the amino acids glutamine and alanine, while the other two strains could not. Strain 07B, which is genetically distinct from the other three strains, exhibited the only nontoxic effect. Thus, molecular tools may support identification, tracking, and prediction of strains and/or ecotypes using distinctive chloroplast gene signatures. “
“This study provides the first morphological features of resting cysts of Cochlodinium polykrikoides collected from Korean coastal sediments. Evidence for the existence of resting cysts of C. polykrikoides is based on the morphological and CYTH4 molecular phylogenetic data of the germinated cells and a resting cyst. The morphology of the resting cysts differed from that reported previously in sediments and culture experiments. The distinct feature is that the cyst body was covered by the reticulate ornaments and spines. “
“The diatoms (Bacillariophyta) from a coastal lagoon from the Diablas wetlands (Isla Isabela, the Galápagos Islands) were studied in material from surface samples and a sediment core spanning the past 2,700 years in order to examine evidence of diatom evolution under geographic isolation. The total number of taxa found was ∼100. Ultrastructural variation in valve morphology between members of Galápagos taxa was used to describe 10 species from the genus Navicula sensu stricto, which are new to science.

4% (VWD 17 9%; platelet function defect 23 2%; mild clotting fact

4% (VWD 17.9%; platelet function defect 23.2%; mild clotting factor deficiencies 3.9%); 11.5% had combined defects. However, 59.6% of these patients had abnormal bleeding of unknown pathogenesis. Prolonged bleeding time (BT) was found as an isolated laboratory abnormality in 18.6% of these patients. Neither differences in bleeding pattern, nor in the relationship between bleeding severity and any haemostatic measurement, Tanespimycin clinical trial were found [45]. A related study in patients with inherited MCB showed that light transmittance aggregometry was highly reproducible if properly standardized. Both normal and abnormal platelet

aggregation in 213 patients were reproducible in 93.3% and 90.4% of the cases respectively [46]; 13.7% of healthy controls had combined abnormalities of platelet aggregation with 10 μm epinephrine and 4 μm ADP. This combination, therefore, was not considered a useful criterion for diagnosing a platelet function disorder [46]. The finding that platelet function defects

were at least as prevalent as VWD, supports the recommendation that an initial laboratory workup should include AZD3965 supplier investigations for both diseases [45]. In a complementary study, the contradictory reports on the influence of gene polymorphisms on platelet function have been addressed. We analysed the genotype–phenotype relationship for six common polymorphisms [ITGB3 1565T>C (HPA-1), GP1BA variable number tandem repeat and 524C>T (HPA-2), ITGA2 807C>T, ADRA2A 1780A>G, and TUBB1 Q43P] in 286 controls and 160 patients with MCB of unknown cause. We found no effect of these polymorphisms on platelet aggregation, secretion, PFA-100® closure times, or thrombin generation in platelet rich plasma. Thus, they appear

to have no impact on platelet function assessed by these commonly employed assays Carbohydrate [47]. Other studies have also identified significant numbers of patients with inherited MCB and no discernible cause, but these observations and their relevance in clinical practice have not been adequately highlighted. Associations of low VWF, platelet function defects and mild clotting factor deficiencies were more frequent than predicted by chance: any combination of these defects occurred in 11.5% of the patients. Combined abnormalities could unmask or increase the bleeding tendency, similar to the multi-factorial risk for thrombosis. Furthermore, the analysis of the BT is also illustrative: 18.6% of patients with bleeding of unknown cause had prolonged BTs; this proportion increased to 39% and 41% in those with platelet function defects and VWD, respectively, and to 55.6% in those with combined abnormalities of VWF and platelet function. These results suggest that low plasma VWF levels, most platelet function defects, and mild to moderate clotting factor deficiencies should be considered risk factors rather than unequivocal bleeding causes.

5) Overall, little interstrain difference in hepatic levels of m

5). Overall, little interstrain difference in hepatic levels of methionine and SAM, or the effect of alcohol feeding (with the exception of a nonsignificant, yet consistent decrease in SAM) was observed (Fig. 5A,B). Liver SAH (Fig. 5C) and homocysteine (Fig. 5E) levels were elevated as a consequence of alcohol feeding in most strains,

with several strains showing a significant effect. Liver SAM/SAH ratios were decreased (Fig. 5D). Liver injury scores were significantly correlated with SAM/SAH ratio (inverse correlation) and liver homocysteine content only when both control and alcohol-fed groups were considered. Plasma hyperhomocysteinemia has been observed in mice but not rats treated intragastrically with an alcohol-containing diet.21 In addition, hyperhomocysteinemia has been associated with the degree of liver injury.27 We observed that plasma levels of homocysteine are elevated in alcohol-fed mice (Fig. 6A) and that the degree of hyperhomocysteinemia is correlated significantly with both overall liver injury (Fig. 6B) and steatosis (Fig. 6C). These correlations remained significant when only alcohol-fed animals were B-Raf inhibition considered (Supporting Table 2). Homocysteine metabolism is dependent on the concordant action by a number of enzymes in the one-carbon metabolism

pathway. To evaluate the mechanisms of interstrain differences in hyperhomocysteinemia, we evaluated the expression of genes or protein levels of major enzymes responsible for the maintenance of the methyl donor pool in the liver (Fig. 7). It has been previously shown that expression of Bhmt is not affected in alcohol-fed C57BL6 mice.21 However, in our study we did observe changes in Bhmt protein in the liver of alcohol-fed mice of some strains (Fig. 7A). There was a significant decreasing nonlinear relationship between alcohol-induced change

in liver Bhmt and plasma homocysteine (Supporting Table 2). Changes in other regulators of one-carbon metabolism were assessed using gene expression, as messenger RNA (mRNA), protein, and activity levels of these enzymes correlate closely.31 Genes encoding 5-methyltetrahydrofolate-homocysteine methyltransferase (Mtr), an enzyme that catalyzes the final step in methionine biosynthesis, and Mthfr, an enzyme that is involved in homocysteine-methionine transition, were CYTH4 generally down-regulated in alcohol-fed mice, especially in strains that exhibited higher liver injury (Fig. 7B,C). Methionine adenosyltransferase 1 alpha (Mat1a), an enzyme that converts methionine into SAM, was markedly induced in strains with low liver injury (Fig. 7D). Glycine N-methyltransferase (Gnmt), an enzyme that converts SAM to SAH, was also induced in strains that had little liver injury and down-regulated in strains that had the most severe injury (Fig. 8A). Similar trends were observed in the expression of adenosylhomocysteinase (Ahcy) (Fig. 8B), cystathionine-beta-synthase (Cbs) (Fig. 8C), and cystathionase (Cth) (Fig.