Gender- and age-matched C57/BL6 wild-type (WT) and ATGL KO litter

Gender- and age-matched C57/BL6 wild-type (WT) and ATGL KO littermates were intraperitoneally injected with 1 mg/kg body weight of a 0.1-mg/mL suspension of tunicamycin (TM) in saline. Mice were reinjected after 24 hours. At 48 hours after the starting point, mice were killed by cervical dislocation. The experimental protocols were approved by the local animal care and use committees according to criteria outlined in the Guide for

the Care and Use of Laboratory Animals prepared by the U.S. National Academy of Sciences (National Institutes of Health publication 86-23, revised 1985). The animals were kindly provided by Rudolf Zechner from the Institute of Molecular Biosciences at Karl-Franzens University (Graz, Austria) Fulvestrant cost and were generated as described previously.26 Animals were fed a standard rodent chow and were housed in a controlled environment with 12-hour light-dark cycles. TM, sodium oleate, and sodium palmitate were from Sigma-Aldrich (Vienna, Austria). Enzymatic assays were used to

measure serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), this website alkaline phosphatase (ALP), cholesterol (CHOL), TGs (Roche Diagnostics, Mannheim, Germany), and free FAs (Wako Chemical, Neuss, Germany). Lipoprotein subfractions were determined by quantitative agarose gel electrophoresis (Helena Biosciences, Gateshead, UK). For conventional light microscopy, livers were

fixed in 4% neutral buffered formaldehyde solution for 24 hours, embedded in paraffin, and stained with hematoxylin and eosin (H&E) or Sirius Red. Frozen tissue was embedded in Tissue Tec (Sakura Finetek Europe B.V., Alphen aan den Rijn, The Netherlands), and sections of why 2 μm were stained with Oil Red O. Double immunofluorescence staining for cleaved caspase-3 and cytokeratin 18 was performed as described previously.30 RNA isolation, complementary DNA synthesis, and real-time polymerase chain reaction (PCR) were performed as described previously.31 All messenger RNA (mRNA) expression data were normalized to 36b4. Oligonucleotide sequences are available upon request. Hepa1.6 cells (American Type Culture Collection, Manassas, VA) were grown in Dulbecco’s modified Eagle’s medium and knockdown for ATGL was performed by using a lentivirus containing a short hairpin RNA against ATGL, as described in the Supporting Materials and Methods. Hepatic nuclei were extracted with the NE-PER Nuclear and Cytoplasmic Extraction kit from Pierce Biotechnology (Rockford, IL). Srebp1c protein levels were determined using the polyclonal antibodiy Srebp-1(K10), sc-367 (Santa Cruz Biotechnology, Santa Cruz, CA).

In clinical practice, plaster fragments may be bonded without har

In clinical practice, plaster fragments may be bonded without harming the accuracy of the final denture, provided that the bonding agent does not cause dimensional alterations. Cyanoacrylate could be a good material because of its ease of use, quick set, wide availability, and low cost. The aim of this study was to assess the dimensional alteration of Type IV plaster fragments bonded with a cyanoacrylate-based adhesive. Materials and Methods: Ten hexagonal regular prisms were made of Type IV plaster, with two reference marks on one of the faces. The distance between the marks was measured under a comparison microscope. After this, the prisms were fractured so that the fracture line would be between the two reference marks, bonded

with a cyanoacrylate-based universal adhesive

and measured again. Results: The mean difference between the measurements performed before and after fracture and bonding of the fragments was 0.0194 mm. At a level of significance of 0.05, there was no statistically significant difference between the measurements before and after fracture and bonding of the dies BMN 673 supplier (p = 0.1582). Conclusion: It may be concluded that bonding of Type IV plaster fragments with a cyanoacrylate-based adhesive did not cause significant dimensional alterations. “
“Purpose: This study evaluated the resistance to corrosion in welds made with Tungsten Inert Gas (TIG) in specimens made of commercially pure titanium (cp Ti) in comparison with laser buy Forskolin welds. Materials and Methods: A total of 15 circular specimens (10-mm diameter, 2-mm thick) were fabricated and divided into two groups: control group—cp Ti specimens (n = 5); experimental group—cp Ti specimens welded with TIG (n = 5) and with laser (n = 5). They were polished mechanically, washed with isopropyl alcohol, and dried with a drier. In the anodic potentiodynamic polarization assay, measurements were taken using a potentiostat/galvanostat in addition to CorrWare software for data acquisition and CorrView for data visualization and treatment.

Three curves were made for each working electrode. Corrosion potential values were statistically analyzed by the Student’s t-test. Results: Statistical analysis showed that corrosion potentials and passive current densities of specimens welded with TIG are similar to those of the control group, and had lower values than laser welding. TIG welding provided higher resistance to corrosion than laser welding. Conclusion: Control specimens welded with TIG were more resistant to local corrosion initiation and propagation than those with laser welding, indicating a higher rate of formation and growth of passive film thickness on the surfaces of these alloys than on specimens welded with laser, making it more difficult for corrosion to occur. “
“Purpose: This study evaluated the bond strength between resin cement and Y-TZP ceramic (Yttrium-stabilized Tetragonal Zirconia Polycrystalline) submitted to different surface conditionings.

Approximately 25% of all samples exceeded the 20 ng/g limit for a

Approximately 25% of all samples exceeded the 20 ng/g limit for aflatoxin B1 (AFB1) adopted by the National Agency for Food and Drug Administration and Control while 83 and 79% of all samples contained AFB1 and total aflatoxins above the European Union limits Wnt inhibitor of 2 and 4 ng/g, respectively. Aflatoxin concentrations in the raw and coated samples were as much as five times higher than those in the roasted and de-coated nuts, respectively. However, no significant difference was recorded between aflatoxin levels in the coated and de-coated samples. This study has shown that roasting of groundnut and testa removal (de-coating) are effective processing interventions that can significantly lower aflatoxin quantities

in the kernels, thus making it Angiogenesis inhibitor fit for human consumption. “
“To analyse the inheritance of fruit ring rot (FRR) resistance and to screen for microsatellite markers linked to resistance/susceptibility, 875 apple hybrid seedlings (Malus domestica, Jonathan × Golden Delicious) were inoculated with five isolates of Botryosphaeria dothidea in 2 years (2008 and 2009). The results indicated that incidence and non-incidence were qualitatively

segregated, and incidence was dominant to non-incidence. The variation in susceptibility within this population was attributed to the segregation of three major genes. For the phenotype of incidence, the severity of lesion development was a quantitative trait. From 230 Cytidine deaminase published microsatellite primer pairs, six markers were identified that were linked to the susceptibility to FRR. CH04d02-120 and Hi08g12-190, located in LG12 and LG2, respectively, were linked to susceptibility to the pathogen isolate Mx1, and their map distances to the susceptibility loci were 8.2 and 5.1 centimorgan (cM), respectively. CH01e01-120 and CH02c02b-100, which were linked to susceptibility to Ls1, were located in LG14 and LG4, and the map distances to the susceptibility loci were 16.9 and 8.4 cM, respectively. CH05d11-150 and CH03a03-230, linked to susceptibility to Lw048, were located in LG12 and LG14; for both of them, the map distance

was 13.4 cM. “
“A series of small-scale controlled inoculation experiments has been conducted during 2005–2009 to determine whether temperature and controlled atmosphere (CA) storage conditions affect significantly the incidence of Botrytis cinerea and Neonectria galligena rots of apples and to assess whether CA regimes can be ‘fine-tuned’ to suppress fungal rotting. The incidence of B. cinerea and N. galligena rots on apple was reduced consistently by storage in lower temperatures (1.5–2°C). In no case was the disease incidence significantly higher than that under air storage conditions. However, the effect of CA conditions on rot development varied greatly from year to year so that overall there were no significant effects of CA conditions on the incidence of rot during storage till the following April.

Disclosures: Gyongyi Szabo – Consulting: Idenix; Grant/Research S

Disclosures: Gyongyi Szabo – Consulting: Idenix; Grant/Research Support: BMS, LDK378 mouse GSK, Cona-tus, Idera, Johnson&Johnson, Novartis, Ocera, Roche, Shering – Plough, Wyeth, Integrated Therapeutics, Idera The following people have nothing to disclose: Michal Ganz, Timea Csak, Shashi Bala, Banishree Saha Background: Them1 is a long-chain fatty acyl-CoA thioesterase that is highly expressed in brown adipose tissue (BAT) and is strongly upregulated by decreasing ambient temperature. At room temperature, Them1 −/− mice exhibit increased energy expenditure and resistance to diet-induced obesity and hepatic steatosis despite increased food consumption (Zhang et al, PNAS, 2012;109:5417). Aim: The

objective of this study was to elucidate the regulatory role of Them1 in energy metabolism, which may contribute in part to the pathogenesis of NAFLD. Methods: Using a temperature controlled Comprehensive Laboratory

Animal Monitoring System (Columbus Tamoxifen solubility dmso Instruments), Them1−/− and Them1+/+ control mice (n = 6/group) were acclimated (48 h) to ambient temperatures ranging from thermoneutrality (30 °C), room temperature (22 °C) and cold (4 °C). This was followed by 48 h of data collection, which included rates of O2 consumption (VO2) and CO2 production (VCO2), physical activity and food intake. Core body temperatures were determined using a rectal probe. Lean body weights were determined by magnetic resonance spectroscopy. Energy expenditure (EE) was calculated using VO2 and VCO2 and adjusted for lean body weights by ANCOVA. Upon completion of experiments, BAT was harvested Axenfeld syndrome to analyze for mRNA expression

levels by qPCR and for ultrastructure analysis by transmission electron microscopy. Results: Cumulative (48 h) values of EE (kJ) were increased when the temperature was reduced from 30 °C to 22 °C, but there were no genotype dependent differences. By contrast, at 4 °C, EE values were higher in Them1 −/− mice (Them1+/+, 154±3; Them1 −/−, 165±2). At 4 °C, Them1 −/− mice were less active (activity counts/48 h; Them1+/+, 80,843±3,570; Them1−/−; 57,561 ±3,386), consumed the same amounts of food (g/g lean body mass; Them1 +/+, 0.73 ± 0.03; Them1 −/−, 0.69 ± 0.01), but tended to lose a greater percentage of their weight (Them1 +/+, 2.2 ± 0.7; Them1 −/−, 3.8 ± 1.2). Core body temperatures did not differ between the two genotypes. Expression of the thermogenic genes Ucp1 and Pgc1α were markedly upregulated in BAT by decreasing ambient temperature, but genotype dependent differences were not observed. In BAT from mice housed at 4 °C, the absence of Them1 was associated with smaller lipid droplets and larger mitochondria. Conclusions: Our data demonstrate that Them1 in BAT functions to suppress thermogenesis potentially by reducing the delivery of fatty acids from lipid droplets to mitochondria for oxidation. Inhibition of Them1 could provide a unique strategy for the management of the obesity and NAFLD. Disclosures: David E.

122 Other approaches have looked at transdifferentiation of hepat

122 Other approaches have looked at transdifferentiation of hepatocytes from other progenitor cell sources. Mesenchymal stem cells,

whether from bone marrow, cord blood, cord lining cells, cord matrix cells, amniotic cells, adipose progenitor and muscle progenitors have been demonstrated to be capable of differentiation into hepatocytes in vitro.110,118 While it would be exhaustive to describe all human transplantation studies to date, a review on liver regeneration would be incomplete if studies relating to the original aims of understanding regeneration are not covered. To date, some 20 human studies have been undertaken, in which attempts were made to enhance liver regeneration. These can be grouped into CH5424802 adult hepatocyte transplantation, fetal hepatocyte transplantation and bone marrow stem cell transplantation. As mature hepatocytes are the main cells that

regenerate the injured liver, roughly 25 patients with acute liver failure Lenvatinib nmr have been transplanted with 107–9 hepatocytes in an attempt to salvage the failing liver.123 Instead of adult hepatocytes, Habibullah et al.124 transplanted 6 patients with acute liver failure with 107 fetal hepatocytes. In these studies, there were transient clinical improvements in encephalopathy and ammonia levels, but there was no overall transplant-free survival benefit. It is likely that the quantity of cells (up to 5% of liver mass) transplanted for each patient may have been too low to register a clinical benefit, and that the window period was too narrow for these cells to regenerate. Although the use

of bone marrow stem cells as candidates for liver regeneration is controversial, the availability of these cells and ease by which they can be harvested has lead to the transplantation of bone marrow stem cells or peripheral blood stem cells in more than 100 tuclazepam patients with cirrhosis. Of note, one small study employed the infusion of AC133+ cells mobilized from the bone marrow after one lobe of the liver has been deliberately embolised, and showed that regeneration in the remaining lobe was augmented.125 Most of these studies are uncontrolled, but clinical improvement in measurable parameters has been claimed.126 The mechanisms by which improvement has occurred are still not known, but studies have shown that remodeling in cirrhotic liver can occur by paracrine signals (metalloproteinases) from bone marrow mesenchymal cells, without actual transdifferentiation into hepatocytes. Whether this work represents true progenitor cell regeneration or the modulation of local environment for native hepatocytes to regenerate, this strategy may yet be promising as long as liver regeneration occurs and clinical outcome is improved.

6C) Furthermore, the levels of RORα protein and pAMPK were well

6C). Furthermore, the levels of RORα protein and pAMPK were well correlated in vivo, as the levels of RORα and pAMPK were decreased

after HFD and the decrease in pAMPK was recovered after adenovirus-mediated expression of RORα (Fig. 6). Recently, Raichur et al. showed that RORα signaling is associated with increased levels of pAMPK in skeletal muscle, which may be related to our observation. 26 Further questions, INK 128 datasheet such as the manner through which RORα activates AMPK, the molecular functions of phosphorylated RORα, and the identification of phosphorylated residues of RORα, need to be investigated in the future. Mutual antagonism

HM781-36B mw between RORα and LXRα has been addressed previously in drug metabolism and metabolic homeostasis. 24, 25 RORα up-regulates the transcriptional expression of Cyp7b1, an enzyme that is critical for the homeostasis of cholesterol, oxysterol, and bile acids, whereas LXRα suppresses the RORα-induced expression of the Cyp7b1 gene. 24, 25 Activity of the LXRα-responsive reporter gene was inhibited by cotransfection with RORα, indicating that LXRα activity is suppressed reciprocally by RORα. Here, we revealed a novel molecular mechanism of RORα-induced repression of the transcriptional function of LXRα. RORα inhibited the autoactivation cycle of transcription of LXRα, thereby decreasing the mRNA level of LXRα. Obviously,

the function of the critical LXRE located on the LXRα promoter was suppressed by RORα, which may be due to the protein–protein interaction between RORα and LXRα (Fig. 3). Additionally, RORα may repress LXR function indirectly, as it activates AMPK, pentoxifylline which inhibits LXRα. 4 The fact that known ligands of LXRα, TO901317 and 24S-hydroxycholesterol, act as inverse agonists of RORα may cause difficulties in interpreting the mutual antagonism mediated by the physical interaction of the receptors. 28, 29 The affinity of ligand–receptor binding and the intracellular availability of specific ligands may determine the mode of this cross-talk. Nevertheless, the efficient down-regulation of the function of LXRα by RORα may provide a valuable tool for restricting many pathological conditions induced by overly functional LXRα, such as hepatic steatosis. The synthetic compounds that down-regulated LXRα via RORα in this study, as well as naturally occurring flavonoids that inhibit LXRα-regulated lipogenic genes, such as naringenin, are good candidates for such therapies (Fig. 7).

Validation of the score in three external cohorts of BCLC C patie

Validation of the score in three external cohorts of BCLC C patients. Methods: This retrospective study included BCLC C HCC patients (n = 160) at diagnosis or during follow-up, treated by chemoembolization (TACE) 27%, sorafenib 30%, TACE and sorafenib 24% and untreated patients 19%. Determining a score based on prognostic variables of our population. Validation

within three external cohorts of BCLC C HCC patients (Rennes, Nancy, Bordeaux). Results: Cirrhosis was viral 45%, alcohol-related 31%, Child-Pugh A 62%, Child-Pugh B 38%. 50% of HCC were infiltrative tumors. The number of nodules was ≥3 in 44% of Cell Cycle inhibitor cases. Portal vein thrombosis was present in 60% of cases, metastasis in 12% of cases. 45% of the patients had elevated AFP ≥ 200 ng / ml at diagnosis. ECOG grade was ≥1 in 85% of cases. Median survival time of Venetoclax chemical structure patients treated by Sorafenib was 7

months [5–10], by TACE: 10 months [6–15], by TACE then Sorafenib: 13 months [10–15], p = 0.462. Multivariate analysis found five prognostic variables associated with overall survival (AFP ng/ml rate at diagnosis, Child-Pugh score, infiltrative vs. encapsulated tumor, nodule number, ECOG grade). These variables were included in a score (N.I.A.C.E) determined from Beta regression coefficients: 1 x (Nodules: 0 if <3, 1 if ≥ 3) + 1.5 x (Infiltrative 0 if no, 1 if yes) + 1.5 x (AFP: 0 if <200, 1 if ≥ 200) + 1.5 x (Child-Pugh: 0 if A, 1 if B) + 1.5 x (ECOG grade 0 if 0, 1 if ≥ 1). With a threshold value <3, the score found two groups with different survival: <3 (n = 38) 16 months [14–27] vs. ≥3 (n = 122) 7 months [6–9], p < .0001 . Application of the

score to three external BCLC C HCC cohorts treated or not by Sorafenib also found two groups with different DNA ligase survival: <3 (n = 37) 10.6 months [4.1–17.1] vs. ≥3 (n = 46) 5.1 months [2.9–7.4] p < .001 Rennes; Score < 3 (n = 28) 16 months [14–25] vs. ≥3 (n = 55) 6 months [4–8] p < .0001 Nancy; <3 (n = 141) 12 months [10–16] vs. ≥3 (n = 236) 6 months [5–7] p < .0001 Bordeaux. Conclusion: This series confirms that BCLC C HCC are a heterogeneous group. We have determined and validated a simple score which can distinguish a sub-group of better prognosis, regardless of treatment. Current treatments do not appear to alter the natural course of BCLC C HCC with a NIACE score >3. Key Word(s): 1.

The extent of hepatic proliferation was comparable to TAK1LPC-KO

The extent of hepatic proliferation was comparable to TAK1LPC-KO mouse livers or livers from partially hepatectomized mice (Supporting Fig. 3B). The observed hyperproliferation in CAIKK2LAP mice could be due to activation of the cell cycle driven by the NF-κB signaling, or by subordinately activated JNK signaling (Supporting Fig. 3C). Sirius-red staining revealed a hepatic fibrosis in 12-week-old

CAIKK2LAP mice (Fig. 2A), whereas no significant fibrosis was seen in 4-week-old CAIKK2LAP mice nor in nontransgenic animals at any age (Fig. 2A; Supporting Fig. 4A). The extent of fibrosis in 12-week-old CAIKK2LAP mice was variable, ranging from mild portal fibrosis (Desmet score 1) to bona fide cirrhosis (Desmet score 4, seen in 2 out of 16 analyzed animals) (control 0.06 ± 0.3, CAIKK2LAP 1.9 ± 1.3, P = 3 × 10−6; Fig. 2B and data not shown). These LY294002 chemical structure data were further confirmed by elevated hydroxyproline content (biochemical marker of collagen deposition; control 1 ± 0.1, CAIKK2LAP 1.8 ± 0.1, P = 3 × 10−8), morphometrical analysis of Sirius-red stained sections, as well as increased hepatic collagen messenger RNA (mRNA) levels (gene Col1a1) in CAIKK2LAP versus control mice (relative to hypoxanthine-guanine see more phosphoribosyltransferase gene [HPRT], control 0.04 ± 0.03, CAIKK2LAP 3.0 ± 2.0, P = 0.002; Fig. 2A,D,E). The higher collagen deposition in CAIKK2LAP mice was PAK5 likely due

to increased HSC activation, given that α-SMA (gene Acta2), an established HSC activation marker, was significantly elevated, both at the mRNA and protein levels (Fig. 2C,E). In addition, we also observed higher levels of several fibrosis-associated genes such as Tgfb1 and Icam (Fig. 2E). Of note, elevated Col1a1 and Acta levels were already seen in 4-week-old CAIKK2LAP mouse livers (Supporting Fig. 4B), suggesting that these mice already display a significant HSC activation, but no appreciable collagen deposition. To study the pathogenesis

of liver fibrosis development in CAIKK2LAP mice, we performed microarray analyses using livers from 4-week-old mice. In all, 1,043 genes were significantly overexpressed in double-transgenic animals compared to controls (Supporting Table 1). Gene ontology analysis revealed hepatocyte stress reaction, inflammation, and chemotaxis as the major pathways altered in CAIKK2LAP animals (Supporting Table 2). The altered expression of selected genes (SAA isoforms Saa1, Saa2, and Saa3; chemokines Ccl2, Ccl5, Cxcl2, Ccl20, and Cxcl10; chemokine receptors Ccr2 and Cxcr4) and the up-regulation of macrophage-related Tnfa, Il6, and Mmp9 was confirmed by quantitative real-time PCR (Fig. 3). Furthermore, elevated SAA, CCL2, and CCL5 serum levels were observed in CAIKK2LAP mice as compared to controls (Fig. 3). To further characterize the hepatic inflammation in CAIKK2LAP mice, we performed immunohistochemical stainings.

19, 55 Risk variants in the NOD2 gene are also associated with sm

19, 55 Risk variants in the NOD2 gene are also associated with small intestinal occurrence of Cohn’s disease, which is mediated by a compromised expression of Paneth cell defensins.10, 22 Here, the lack of Paneth cell function, especially by the α-defensins HD5 and HD6,10, 22 leads to decreased mucosal

killing activity and shift in the bacterial composition, Palbociclib which likely facilitates bacterial overgrowth and translocation and secondary mucosal and systemic inflammation. In summary, in the healthy state a complex interplay between commensal microbes and the intestinal mucosa results in the establishment of a delicate balance and an intact barrier against pathogens, which prevents inflammation.56 In view of the findings reported here, we hypothesize that in predisposed animals liver cirrhosis leads to the disruption of this host–bacteria balance, mediated by relative deficiency of Paneth cell defensins, particularly in the ileum. This defect weakens intestinal antimicrobial defenses, may lead to progressive changes in the composition of the intestinal microbiota,57-59 and promotes BT. This novel aspect of disease pathophysiology suggests that therapeutic strategies should aim at restoring the host mucosal barrier in cirrhosis. Finally, the data presented emphasize the presence of the liver–gut axis,

underscoring its bidirectional communication. Additional Supporting Information may be found in the online version of this article. “
“Bile acid metabolism is intimately linked to the control of energy homeostasis and glucose and lipid metabolism.

The nuclear receptor farnesoid X receptor (FXR) plays a major role Cilomilast in the enterohepatic cycling of bile acids, but the impact of nutrients on bile acid homeostasis is poorly characterized. Metabolically active hepatocytes cope with increases in intracellular glucose concentrations by directing glucose into storage (glycogen) or oxidation (glycolysis) pathways, as well as to the pentose phosphate shunt and the hexosamine biosynthetic pathway. Here we studied whether the glucose nonoxidative hexosamine biosynthetic pathway modulates FXR activity. Our results show that FXR interacts with and is O-GlcNAcylated by O-GlcNAc transferase in its N-terminal AF1 domain. Increased FXR O-GlcNAcylation enhances FXR gene expression and protein stability Morin Hydrate in a cell type-specific manner. High glucose concentrations increased FXR O-GlcNAcylation, hence its protein stability and transcriptional activity by inactivating corepressor complexes, which associate in a ligand-dependent manner with FXR, and increased FXR binding to chromatin. Finally, in vivo fasting-refeeding experiments show that FXR undergoes O-GlcNAcylation in fed conditions associated with increased direct FXR target gene expression and decreased liver bile acid content. Conclusion: FXR activity is regulated by glucose fluxes in hepatocytes through a direct posttranslational modification catalyzed by the glucose-sensing hexosamine biosynthetic pathway.

Because of the large proportion of Medicare patients that enter t

Because of the large proportion of Medicare patients that enter the program in advanced stages of disease, treatment prior to Medicare entry is likely to be more effective in mitigating the health consequences of HCV. Disclosures: David B. Rein – Grant/Research Support: Gilead Selleckchem PS 341 Sciences, Inc. The following people have nothing to disclose: John S. Wittenborn, Danielle Liffmann, Joshua M. Borton Background: Several combinations of Direct Acting Antivirals (DAAs) can cure Hepatitis C (HCV) in the majority of treat-ment-naïve patients, in a range of genotypes. Mass treatment programmes to cure HCV in developing countries are only feasible if the costs of treatment and monitoring are very low.

This analysis aimed to estimate minimum costs of DAA treatment and associated diagnostic monitoring. Methods: Clinical trials of HCV DAAs were reviewed to identify combinations with consistently high rates of Sustained Virological Response (SVR) in different genotypes. For each DAA, molecular structures, doses, treatment duration and components of retro-synthesis were used to estimate costs of mass production. Manufacturing

costs per gram of DAA were projected as formulated product cost, based upon treating at least 5 million patients/year (to arrive at volume demand) and a 40% margin for formulation. Costs of diagnostic support were estimated based on published developing country prices of genotyping, HCV antigen tests (to confirm infection pre-treatment and identify relapse/re-infection post-treatment), AZD8055 price plus full blood count/clinical chemistry.

Avelestat (AZD9668) Results: Predicted minimum costs for 12-week courses of HCV DAAs (patent expiry dates) were: US$50 for ribavirin 1200mg/day (generic), US$20 for daclatasvir 60mg/day (2027), US$102 for sofosbuvir 400mg/day (2029), US$90 for ledipasvir 90mg/day (2030), US$44 for MK-8742 (2028), and US$71 for MK-5172 (2030). Predicted minimum costs for 12 week courses of combination DAAs with the most consistent efficacy results were: US$122 per person for sofosbuvir+da-clatasvir, US$152 for sofosbuvir+ribavirin (US$304 for 24 weeks), US$192 for sofosbuvir+ledipasvir and US$115 for MK-8742+MK-5172. Diagnostic testing costs were estimated at US$90 for genotyping (if treatment not pan-genotypic), US$34 for two HCV antigen tests (lower detection limit 2000 IU/mL) and US$22 for two full blood count, ALT and creati-nine tests (before and during treatment). Conclusions: Minimum costs of treatment and diagnostics to cure HCV were estimated at US$171-360 per-person, without genotyping or US$261-450 per-person with genotyping. These cost estimates assume that similar large-scale treatment programmes for HIV/AIDS can be established for HCV. Treatments with proven pan-ge-notypic activity will be required to avoid expensive pre-treat-ment genotyping. Further reductions in price could be achieved through shorter durations of treatment, if efficacy is shown in future trials. Disclosures: Andrew M.