6-8 Nevertheless, correlations are not always found between tumor stage and the actual prognosis, and this phenomenon is more common in patients with early HCC than in those with advanced HCC. Considering that HCC is increasingly diagnosed and resected at an early stage and that current staging systems have some limitations in the prognostic evaluation of early HCC,4, 9, 10 efforts have been made to investigate prognostic molecules.11-13 To date, no easily measurable biomarker that has strong correlations with

clinical outcomes has been identified. Human aspartyl-(asparaginyl)-β-hydroxylase (AAH) is abundantly expressed in proliferating trophoblastic cells of the placenta and is rarely expressed in normal adult tissues.14 Overexpression of AAH can promote the malignant transformation of biliary epithelial cells, enhance the metastasis of cholangiocarcinoma cells,14-17 3-MA molecular weight and increase the mobility of neuroblastoma.18 Wands and colleagues14, 19 have reported that AAH is highly expressed in HCCs and that overexpression of AAH significantly increases motility and invasiveness of HepG2 and Huh-7 cells.20, 21 Our previous study also showed enhanced AAH immunostaining in HCC when compared with that in nontumorous tissue, which was associated with poor differentiation of HCC cells.22 However, the U0126 nmr significance of AAH expression level in the prognostic evaluation of patients undergoing surgical resections

has not been reported. The purpose of this prospective study was to examine whether AAH expression level is an effective prognostic factor for HCC patients who undergo hepatectomy. Pembrolizumab ic50 Differential expression levels of AAH in HCCs and nontumorous tissues were analyzed through hybridization with complementary DNA (cDNA) microarray, reverse-transcription polymerase chain reaction (RT-PCR), and immunohistochemical staining in tissue microarray (TMA). We found that AAH expression level in tumor tissue was well-correlated with multiple malignant clinico-pathological

characteristics and was strongly associated with tumor recurrence and patient survival. Moreover, AAH overexpression predicted a worse surgical outcome in patients with early stage HCC according to Barcelona Clinic Liver Cancer (BCLC) staging classification. AAH, aspartyl-(asparaginyl)-β-hydroxylase; AFP, AFP, α-fetoprotein; BCLC, Barcelona Clinic Liver Cancer; cDNA, complementary DNA; CI, confidence interval; HCC, hepatocellular carcinoma; HR, hazard ratio; mRNA, messenger RNA; PVTT, portal vein tumor thrombosis; RT-PCR, reverse-transcription polymerase chain reaction; TMA, tissue microarray; TTR, time to recurrence. A total of 281 adult patients with HCC undergoing hepatectomy by three independent surgical teams at Eastern Hepatobiliary Surgery Hospital between January 1 and June 30, 2004, were enrolled in the study. The preoperative clinical diagnosis of HCC met the diagnostic criteria of the American Association for the Study of Liver Diseases.

Moreover, a lysine acetylation/deacetylation-sumoylation switch has been implicated in the functional regulation of several important molecules.22, 23 Ethanol inhibits sirtuin 1 (SIRT1), an nicotinamide adenine dinucleotide-positive–-dependent class III protein deacetylase, both in cultured hepatic cells and in animals.13, 24 It is possible that this ethanol-mediated hyperacetylation/hyposumoylation of lipin-1 may be a consequence of the inhibition of SIRT1 by ethanol. Whether acetylation/sumoylation would serve as a molecular switch to control the nuclear localization and coactivator activity of lipin-1 in the liver and how ethanol affects the functional relationship of SIRT1 and lipin-1

selleck compound are currently under investigation in our laboratory. Lipin-1 localizes to the nucleus and is a component of a transcriptional complex with PPARα/PGC-1α, which stimulates fatty acid oxidation in the liver.3 Ethanol-mediated dysregulation of the hepatic PPARα/PGC-1α axis and subsequent incomplete stimulation of PPARα/PGC-1α target genes involved in fatty acid oxidation contributes to the development of alcoholic liver steatosis.17 Taken together with a recent study demonstrating that a high-fat-diet–induced fatty liver

is partially mediated by impairment of the PGC-1α/nuclear lipin-1/PPARα axis and fatty acid oxidation in mice, our current findings suggest that depletion of nuclear lipin-1 is likely to lead to impairment of the Idasanutlin supplier PPARα/PGC-1α axis and fatty acid oxidation in the livers of chronically ethanol-fed animals.25 Furthermore, lipin-1 subcellular localization regulates SREBP-1 signaling and Glycogen branching enzyme governs the

assembly and secretion of very-low-density lipoproteins (VLDLs).1, 4 It is tempting to speculate that ethanol-induced nucleocytoplasmic shuttling may activate SREBP-1 and impair VLDL secretion and, subsequently, contribute to hepatic fat accumulation. Another major novel finding of the present study is that ethanol up-regulates lipin-1 largely through inhibition of AMPK and activation of SREBP-1. Our study provides evidence, for the first time, to our knowledge, that AMPK is involved in the regulation of lipin-1 gene expression. However, the exact mechanism by which AMPK inhibition by ethanol leads to activation of SREBP-1, and subsequent inhibition of Lpin1, remains to be determined. Our earlier work showed that ethanol selectively increases hepatic SREBP-1 activity in rodent models through inhibition of AMPK.6, 9 AMPK directly phosphorylates SREBP-1 and suppresses SREBP-1 activity in hepatocytes exposed to high glucose.26 Conceivably, ethanol-mediated inhibition of AMPK may cause reduced phosphorylation of SREBP-1, which, in turn, results in activation of proteolytic processing and transcriptional activity of SREBP-1 and, ultimately, increased lipin-1 gene expression. Moreover, several lines of evidence have suggested functional connections between SIRT1 and AMPK.

We have previously demonstrated that sylvanus and cynomolgus macaques are susceptible to in vivo HBV infection after intrahepatic HBV DNA inoculation.

In this study, we evaluated the susceptibility of primary macaque hepatocytes (PMHs) to HBV infection with a highly efficient HBV genome–mediated transfer system via a recombinant baculovirus (Bac-HBV). Freshly prepared PMHs, isolated from macaque liver tissue by collagenase Selleck Rapamycin perfusion, were transduced with Bac-HBV, and intermediates of replication were followed for 9 days post-transduction. Evidence of HBV replication (hepatitis B surface antigen secretion, viral DNA, RNA, and covalently closed Caspase-dependent apoptosis circular DNA) was detected from day 1 to day 9 post-transduction. HBV markers were dose-dependent and still detectable at a multiplicity of infection of 10. Importantly, transduced PMHs secreted all typical forms of HBV particles, as evidenced by a cesium chloride gradient as well as transmission electron microscopy. Furthermore, the Toll-like receptor 9 (TLR9) ligand was used to stimulate freshly prepared macaque peripheral blood mononuclear cells to generate TLR9-induced

cytokines. We then demonstrated the antiviral effects of both TLR9-induced cytokines and nucleoside analogue (lamivudine) on HBV replication in transduced PMHs. Conclusion: Baculovirus-mediated genome transfer initiated a full HBV replication cycle in PMHs; thus highlighted both the baculovirus efficiency in crossing the species barrier and macaque susceptibility

to HBV infection. Moreover, our results demonstrate the relevance of thus system for antiviral compound evaluations with either nucleoside analogues or inhibitory cytokines. Cynomolgus macaques are readily available, are immunologically closely related to humans, and may therefore represent for a promising model for the development of new immunotherapeutic strategies. (HEPATOLOGY 2010) Despite an effective vaccine, hepatitis B virus (HBV) infection remains a major public health problem because more than 350 million people are chronic HBV carriers worldwide and have a greater risk of developing severe liver diseases such as cirrhosis and hepatocellular carcinoma.1 Currently available therapies are restricted to the use of standard/pegylated interferon-alpha (IFN-α) or nucleos(t)ide analogues such as lamivudine, but both are still unsatisfactory. Indeed, despite its immunomodulatory and antiviral properties, IFN-α is effective in less than 30% of chronic carriers, and severe adverse side effects restrict its use. In contrast, nucleos(t)ide analogues are well tolerated and strongly suppress viral replication, but they require indefinite therapy, which leads to the emergence of drug-resistant mutants.

The nuclear and cell boundaries could not be seen clearly. Moreover, many dead cells or cell debris with bright green fluorescence were floating Atezolizumab clinical trial above the living cell layer. These phenomena suggest that G2-122×8 cells might have been undergoing apoptosis or/and differentiation. To determine whether miR-122 promoted hepatocyte differentiation, we quantified the mRNA expression of three cytochrome P450 family genes (CYP1A2, CYP2C9, and

CYP7A1) that are hepatic functional proteins specifically expressed in mature hepatocytes.18, 27, 28 Notably, CYP7A1 is a known target of CUTL1 in HepG2 cells.27 As shown in Fig. 5F, in G2-122×4 cells, only the expression of CYP7A1 increased, Selleck MAPK Inhibitor Library whereas in G2-122×8 (B7) cells, all three cytochrome P450 genes were significantly up-regulated. This result indicates that the continuous high level of miR-122 eventually induces the differentiation of hepatoblastoma cells. In addition,

it suggests that CUTL1 is an important functional target of miR-122. In combination, our studies suggest that the activation of miR-122 plays an important role in guiding hepatocyte differentiation during development. This study demonstrates that four LETFs (C/EBPα, HNF1α, HNF3β, and HNF4α) are involved in the transcriptional regulation of miR-122, which could directly regulate a group of target genes involved in proliferation and differentiation regulation. In line with this, restoration of miR-122 in hepatoblastoma cells suppresses cellular proliferation and activates the expression of hepatocyte functional genes. We show that CUTL1 is a biological target of miR-122 during liver development. Our findings support a role of miR-122 in liver development, as shown in Fig. 6. According to this model, miR-122 acts as an important bridge connecting the two different types of regulators that control the balance between the proliferation and differentiation of hepatocytes IKBKE during liver development. The transcriptional regulation of the majority of miRNAs is currently unknown.

Because miR-122 is the most abundant and specific miRNA in the liver, clarification of its regulatory mechanism is necessary to reveal the transcriptional regulation of liver miRNAs. Here, we provide the first direct evidence that the transcription of miR-122 is regulated by several LETFs. The involvement of several transcription factors in the transcriptional regulation of a single miRNA has not been reported previously. However, this mechanism is a common principle for liver gene regulation.17, 18 While our study was underway, others identified LETFs as central regulatory molecules in gene networks associated with the loss of miR-122 in human HCCs, and their knockdown experiments suggest that miR-122 is under the transcriptional control of HNF1α, HNF3α, and HNF3β.

PKR, eIF2α and JNK1 phosphorylation/activation were studied by western blots from liver tissues. Apoptosis was assessed by the active caspase 3 subunit. Fibrosis tested by procollagen a1(I) ASMA, TGFβ expression and hydroxyl proline assay. In vitro, primary wt hepatocytes were exposed to palmitate and NOX4 promoter studies were conducted. Wt hepatocytes were transfected with the PKR siRNA, exposed to palmitate and eiF2a and JNK1 phosphorylation were evaluated. selleck chemicals Results: NO X4 expression was induced in the fl/fl

mice on the CDAA diet (p < 0.05). The TG content significantly improved in the Albcre/NOX4 mice (p<0.01) with the downregulation of SREBP1c, Fas (p = 0.04), and HIF inhibitor PPARγ compared to fl/fl controls. PKR, eIF2α, JNK1

and CHOP activation were significantly attenuated in the Albcre NOX4 mice on the CDAA diet. Hepatocyte apoptosis (active caspase 3) and fibrosis also improved significantly in the NOX4/Albcre mice with lower procollagen α1(I) ASMA, TGFβ expression and hydroxy proline content (p < 0.05). In vitro, palmitate induced NO X4 promoter activity. PKR, eiF2α, JNK1 phosphorylation, and CHOP activation have decreased in the palmitate-treated NOX4-/- hepatocytes. In the PKR siRNA and palmitate treated fl/fl cells JNK1 and eiF2α induction were significantly attenuated. Conclusion: NO X4 induction in hepatocytes by saturated fatty acids contributes to steatosis and to the induction of PKR-mediated stress pathways and cell death during NASH progression. Targeting NO X4 could thus be beneficial to improve hepatocyte injury and fibrosis in NASH. Disclosures: The following people have nothing to disclose: Tzu-I Chao, Xiaosong Jiang, Hiroo Fukada, Fawaz Haj, Ahmed Bettaieb,

Natalie Torok Background: Non-alcoholic steatohepatitis (NASH) is characterized by fat accumulation, chronic inflammation (includingmacrophages) DNA ligase and fibrosis that can lead to cirrhosis or hepatocellular carcinoma. Treatment options are very limited. Recent evidence suggests that C-C chemokine receptor (CCR) type 2 and its main ligand, monocyte chemotactic protein-1, contribute to macrophage recruitment in the liver. Cenicriviroc (CVC) is an oral, potent, dual CCR2/CCR5 antagonist that showed favorable safety and tolerability over 24 weeks of dosing in an ongoing 48-week Phase 2b study in 143 HIV-1-infected adults (NCT01338883). We evaluated CVC in a mouse model of NASH that leads to hepatocellular carcinoma; data from the first, fibrotic stage of the model are presented. Methods: NASH was induced in male mice by a single injection of 200μg streptozotocin 2 days after birth (causing insulin resistance), followed by a high fat diet from 4 weeks of age.

PKR, eIF2α and JNK1 phosphorylation/activation were studied by western blots from liver tissues. Apoptosis was assessed by the active caspase 3 subunit. Fibrosis tested by procollagen a1(I) ASMA, TGFβ expression and hydroxyl proline assay. In vitro, primary wt hepatocytes were exposed to palmitate and NOX4 promoter studies were conducted. Wt hepatocytes were transfected with the PKR siRNA, exposed to palmitate and eiF2a and JNK1 phosphorylation were evaluated. FDA approved Drug Library high throughput Results: NO X4 expression was induced in the fl/fl

mice on the CDAA diet (p < 0.05). The TG content significantly improved in the Albcre/NOX4 mice (p<0.01) with the downregulation of SREBP1c, Fas (p = 0.04), and Ibrutinib mw PPARγ compared to fl/fl controls. PKR, eIF2α, JNK1

and CHOP activation were significantly attenuated in the Albcre NOX4 mice on the CDAA diet. Hepatocyte apoptosis (active caspase 3) and fibrosis also improved significantly in the NOX4/Albcre mice with lower procollagen α1(I) ASMA, TGFβ expression and hydroxy proline content (p < 0.05). In vitro, palmitate induced NO X4 promoter activity. PKR, eiF2α, JNK1 phosphorylation, and CHOP activation have decreased in the palmitate-treated NOX4-/- hepatocytes. In the PKR siRNA and palmitate treated fl/fl cells JNK1 and eiF2α induction were significantly attenuated. Conclusion: NO X4 induction in hepatocytes by saturated fatty acids contributes to steatosis and to the induction of PKR-mediated stress pathways and cell death during NASH progression. Targeting NO X4 could thus be beneficial to improve hepatocyte injury and fibrosis in NASH. Disclosures: The following people have nothing to disclose: Tzu-I Chao, Xiaosong Jiang, Hiroo Fukada, Fawaz Haj, Ahmed Bettaieb,

Natalie Torok Background: Non-alcoholic steatohepatitis (NASH) is characterized by fat accumulation, chronic inflammation (includingmacrophages) Nintedanib (BIBF 1120) and fibrosis that can lead to cirrhosis or hepatocellular carcinoma. Treatment options are very limited. Recent evidence suggests that C-C chemokine receptor (CCR) type 2 and its main ligand, monocyte chemotactic protein-1, contribute to macrophage recruitment in the liver. Cenicriviroc (CVC) is an oral, potent, dual CCR2/CCR5 antagonist that showed favorable safety and tolerability over 24 weeks of dosing in an ongoing 48-week Phase 2b study in 143 HIV-1-infected adults (NCT01338883). We evaluated CVC in a mouse model of NASH that leads to hepatocellular carcinoma; data from the first, fibrotic stage of the model are presented. Methods: NASH was induced in male mice by a single injection of 200μg streptozotocin 2 days after birth (causing insulin resistance), followed by a high fat diet from 4 weeks of age.

TDF was well tolerated overall; 3 patients (1.1%) discontinued the study early for an AE (2-MRI and 1-NRF). No patients had a confirmed increase in serum creatinine of ≥0.5 mg/dL, and 1% (2-NRF) had transient PO4 <2 mg/dL. Nine MRI patients had CLCr <50 mL/min (pre-treatment range: 49–61 mL/min) that stabilized with dose adjustment. No differences were observed in % change in spine or hip BMD over 96 weeks, and no clinically relevant bone loss was noted in either group. At Week 96 there was no significant difference (missing = failure) in % with HBV DNA < 400 copies/mL, or rates of ALT normalization or HBeAg loss/seroconversion. Conclusions: The

safety, PK, and efficacy of patients with MRI receiving TDF were similar to NRF patients; in MRI patients there was no evidence of increased

risk for renal- or bone-related complications. W SIEVERT,1 S STRASSER,2 E GANE,3 J GEORGE,4 F WEILERT,5 signaling pathway JF FLAHERTY,6 P DINH,6 KM KITRINOS,6 JG MCHUTCHISON,6 P MARCELLIN7 1Monash University and Monash Medical Centre, Melbourne, VIC; 2Royal Prince Alfred Hospital, Cell Cycle inhibitor Sydney, NSW; 3Auckland City Hospital, Auckland, New Zealand; 4Storr Liver Unit, University of Sydney at Westmead Hospital, Sydney, NSW; 5Waikato Hospital, Hamilton, New Zealand; 6Gilead Sciences, Foster City, CA, USA; 7Hôpital Beaujon, Clichy, France. Background: We previously reported that 5 years of tenofovir DF (TDF) therapy in mostly treatment naïve patients results in sustained virological suppression with no development

of resistance and was associated with either the halting or regression of fibrosis in 96% of patients. Here we present 6 year results from these two ongoing 8 year studies (Studies 102 and 103). Methods: After 48 weeks of double-blind comparison of TDF to adefovir dipivoxil, Fludarabine solubility dmso all patients undergoing liver biopsy were eligible to continue open-label TDF. Patients were assessed every 3 months for safety and efficacy with annual resistance surveillance; annual assessments of bone mineral density (BMD) of the spine and hip by DXA were added starting at year 4. Results: In a total 641 patients who were initially randomized and treated, 585 (93%) entered the TDF extension phase, and at Year 6, 466 (73%) remain on study. Efficacy results at Year 6 are shown in the table. TDF was well tolerated over the 6 year evaluation period. Less than 2% of patients discontinued TDF due to an adverse event, and ≤1.5% experienced a confirmed renal event (≥0.5 mg/dL increase in serum creatinine from baseline, phosphorus <2 mg/dL, or CrCL <50 mL/min). BMD (T scores) was stable over 2 years of evaluation. No resistance to TDF has been detected through Year 6. Conclusions: In these two trials, TDF remains safe and effective over a 6 year treatment period, with no detectable resistance to TDF; a relatively low rate of renal events and no evidence of clinically relevant bone loss were also observed.

The selection of optimal cut-off point values

was based on the IL-22, HBsAg and HBcrAg levels at which accuracy was maximal. Optimal cut-off value, sensitivity, specificity, positive predictive value, negative predictive value and calculated area under the curve (AUC) values for each parameter are listed in Table 5. The AUC values were consistently high and ranged between 0.731 (IL-22) and 0.858 (HBcrAg). Several factors found in association with a VR to ETV therapy were evaluated for their independence by multivariate analysis. We determined that IL-22 of 27.8 pg/mL or more (hazard ratio [HR] = 13.67 [95% confidence interval [CI] = 1.05–178.11], P = 0.046) and HBcrAg of 5.7 log U/mL or less (HR = 10.88 [95% CI = 1.02–115.44], P = 0.048) were independent factors related to a Selleck Y 27632 VR. HBsAg did

not have a significant Vemurafenib independent association in this study (P = 0.071). Longitudinal analysis of IL-22, HBsAg and HBcrAg levels was carried out at 6, 12 and 24 months after the initiation of therapy and showed significant gradual reductions in IL-22 (P < 0.001, Friedman test), HBsAg (P < 0.001) and HBcrAg (P < 0.001) in samples collected from patients who achieved a VR (Fig. 1). We noted a higher median serum IL-22 concentration at month 6 in the VR group than in the non-VR group (P = 0.012), and there were significant differences at each time point for HBsAg (6 months, P = 0.002; 12 months, P = 0.006; and 24 months, P = 0.004) and HBcrAg (6 months, P < 0.001; 12 months, P < 0.001; and 24 months, P < 0.001) between responders and non-responders. In the present study, we measured the levels of six cytokines and five chemokines in patients with chronic hepatitis B and analyzed their association with ETV therapy outcome using a bead-array multiplex immunoassay system. mafosfamide Four of our observations are noteworthy and require further comment. First, serum IL-6, CCL2,

CXCL9 and CXCL10 concentrations were higher in patients with chronic hepatitis B than in healthy subjects. Second, serum IL-22 concentration before treatment was significantly higher in patients achieving a VR to ETV therapy. In contrast, responders had lower serum levels of HBsAg and HBcrAg at baseline. Third, IL-22, HBsAg and HBcrAg decreased during treatment and remained low in patients with a VR. Fourth, serum IL-6, CXCL9, CXCL10 and CXCL11 were positively correlated with serum values of AST, ALT and bilirubin, but were negatively correlated with HBsAg. Interleukin-6 is a well-recognized multifunctional cytokine that may reflect more active hepatic necroinflammation and be associated with chronic HBV infection severity.

The selection of optimal cut-off point values

was based on the IL-22, HBsAg and HBcrAg levels at which accuracy was maximal. Optimal cut-off value, sensitivity, specificity, positive predictive value, negative predictive value and calculated area under the curve (AUC) values for each parameter are listed in Table 5. The AUC values were consistently high and ranged between 0.731 (IL-22) and 0.858 (HBcrAg). Several factors found in association with a VR to ETV therapy were evaluated for their independence by multivariate analysis. We determined that IL-22 of 27.8 pg/mL or more (hazard ratio [HR] = 13.67 [95% confidence interval [CI] = 1.05–178.11], P = 0.046) and HBcrAg of 5.7 log U/mL or less (HR = 10.88 [95% CI = 1.02–115.44], P = 0.048) were independent factors related to a Z-VAD-FMK ic50 VR. HBsAg did

not have a significant AP24534 independent association in this study (P = 0.071). Longitudinal analysis of IL-22, HBsAg and HBcrAg levels was carried out at 6, 12 and 24 months after the initiation of therapy and showed significant gradual reductions in IL-22 (P < 0.001, Friedman test), HBsAg (P < 0.001) and HBcrAg (P < 0.001) in samples collected from patients who achieved a VR (Fig. 1). We noted a higher median serum IL-22 concentration at month 6 in the VR group than in the non-VR group (P = 0.012), and there were significant differences at each time point for HBsAg (6 months, P = 0.002; 12 months, P = 0.006; and 24 months, P = 0.004) and HBcrAg (6 months, P < 0.001; 12 months, P < 0.001; and 24 months, P < 0.001) between responders and non-responders. In the present study, we measured the levels of six cytokines and five chemokines in patients with chronic hepatitis B and analyzed their association with ETV therapy outcome using a bead-array multiplex immunoassay system. Methisazone Four of our observations are noteworthy and require further comment. First, serum IL-6, CCL2,

CXCL9 and CXCL10 concentrations were higher in patients with chronic hepatitis B than in healthy subjects. Second, serum IL-22 concentration before treatment was significantly higher in patients achieving a VR to ETV therapy. In contrast, responders had lower serum levels of HBsAg and HBcrAg at baseline. Third, IL-22, HBsAg and HBcrAg decreased during treatment and remained low in patients with a VR. Fourth, serum IL-6, CXCL9, CXCL10 and CXCL11 were positively correlated with serum values of AST, ALT and bilirubin, but were negatively correlated with HBsAg. Interleukin-6 is a well-recognized multifunctional cytokine that may reflect more active hepatic necroinflammation and be associated with chronic HBV infection severity.

Smooth LPS from strain C28 did not cause leakage of K+ or of UV-absorbing Venetoclax datasheet material and did not prevent growth of C28. The relevance of these findings is discussed in relation to disease. “
“One hundred and eighty isolates of Rhizoctonia solani AG1-IA, the causal agent of rice sheath blight,

were obtained from six locations in southern China. The genetic structure of R. solani isolates was investigated using random amplified polymorphic DNA (RAPD) markers, and a considerable genetic variation among R. solani isolates was observed. Most of the genetic diversity was distributed within populations, rather than among them. The distribution pattern of the genetic variation of R. solani appears to be the result of high gene flow (Nm) and low-genetic differentiation among populations. The aggressiveness of R. solani was visually assessed by rice seedlings of five different cultivars in the glasshouse. All isolates

tested were found to induce significantly different levels of disease severity, reflecting considerable variation in aggressiveness. The isolates were divided into highly virulent, moderately virulent and weakly virulent groups, and the moderately virulent isolates were dominant in R. solani U0126 cost population. No significant correlation was observed among the genetic similarity, pathogenic aggressiveness and geographical origins of the isolates. Information obtained from this study may be useful for breeding for improved resistance to sheath blight. “
“Wheat powdery

mildew caused by Blumeria graminis f.sp. tritici (Bgt) is an important and destructive disease worldwide. Detection of latent infection of wheat seedlings is critical to estimate initial inoculum potential of epidemics in the fields. To improve the conventional method, a nested PCR approach had been established in this study to detect latent infections of wheat leaves caused by Bgt. The DNA primer sets including external and internal primer pairs for the nested PCR were designed followed by testing their specificities to Bgt by using Bgt and other fungal species of wheat. Sensitivity test demonstrated that the nested PCR could detect as low as 0.1 fg Buspirone HCl template DNA and about 10,000 times more sensitive than the standard PCR. Results of artificial inoculation experiments showed that the nested PCR assay can detect a low level of latent infection of wheat seedlings 2 days earlier than did standard PCR. The incidences of latent infection of wheat seedlings determined by the nested PCR linearly correlated with those by the conventional incubation method (r2 = 0.66, P = 0.0023). The incidences of latent infection detected with nested PCR were higher than that with the conventional method. This study provides an accurate method to efficiently estimate the initial inoculum potential of wheat powdery mildew epidemics in the fields. “
“Colletotrichum spp.