Alternatively, serum-induced hepatic differentiation was performed as described.22 For reverse-transcription polymerase chain reaction (RT-PCR), total RNA was extracted using RNeasy mini-kits and then treated with RNase free DNase (Qiagen, Valencia, CA). One microgram of total RNA was then reverse-transcribed to complementary DNA using a Superscript RT kit (Invitrogen,

Carlsbad, CA) with random hexamers. PCR was performed using Taq polymerase (Takara Bio, Shiga, Japan) in PCR buffer containing 2.5 mM MgCl2 and 0.2 μM dNTPs. Oligonucleotide primers were performed ACP-196 concentration using the primer pairs shown in Supporting Table 1. For real-time PCR, commercially available assay mixes for Alb, Afp, carbamoyl phosphate synthetase (Cps1), transcription factor 1 (Tcf1), Hex, BMP-4, Delta-like 1 (Dlk1), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were used to quantify messenger RNA (mRNA) levels, and PCR was performed using a Prism 7700 Sequence Detector (Applied Biosystems, Foster City, CA). mRNA levels were normalized to GAPDH mRNA levels in the same samples. EBs-derived cells were stained with a PE-conjugated anti-c-kit antibody (BD PharMingen, San Diego, CA), after which the cells were analyzed using a FACSan (Becton Dickenson, San Jose, CA) or sorted on a FACS Aria cell sorter (Becton Dickenson). Foxa2 staining of brachyury+ and c-kit+ cells was performed in microtiter wells as described.22

Briefly, the cells were incubated with an anti-Foxa2 Palbociclib manufacturer primary antibody

(goat polyclonal P-19; Santa Cruz Biotechnololgy, Santa Cruz, CA) and visualized using Cy3-conjugated anti-goat secondary immunoglobulin G antibody (Jackson Immunoresearch, West Grove, PA). For Alb staining, day 14 EBs were scraped, embedded in Tissue Tek O.C.T. compound (Sakura, Torrance, CA), and frozen in liquid nitrogen, after which 4-μm-thick sections were cut on a cryostat and placed on polylysine-coated glass microscope slides. After the fixation and permeabilization, the cells were incubated for 1 hour with anti-Alb primary antibody (Biogenesis, Kingston, NH) and visualized using a Cy3-conjugated anti-rabbit immunoglobulin G secondary antibody (Jackson Immunoresearch). After culturing EBs for 14 days under various conditions, the medium was changed to serum-free Iscove’s modified Dulbecco’s medium containing 2 mM glutamine. The EBs were then incubated for Fludarabine an additional 24 hours, and the conditioned medium was collected for assay. Alb and transferrin concentrations in the conditioned medium were measured using solid-phase sandwich enzyme-linked immunosorbent assays (Bethyl, Montgomery, TX) according the manufacturer’s instructions. For microarray analysis, total RNA was extracted using RNeasy mini kits (Qiagen), after which 10 μg of fragmented target total RNA was used for hybridization of each UniSet Mouse I Expression Bioarray chip (Amersham Life Sciences, Buckinghamshire, UK), which contained 10,012 probes.

This variant is not known to confer reduced susceptibility to narlaprevir. All patients with treatment-emergent resistance variants failed to achieve undetectable viral HCV-RNA levels. Virological breakthrough was observed HSP mutation in four patients; one previous nonresponder appeared to be a nonresponder again during SOC. One treatment-experienced patient with a serine-54 polymorphism at baseline associated with reduced susceptibility to narlaprevir achieved undetectable viral load levels in period 2

(cohort 2). This patient remained HCV-RNA undetectable during SOC but relapsed after 24 weeks of treatment. No severe or serious adverse events (AEs) and no dosing interruptions or discontinuations were reported during narlaprevir dosing. A complete listing of the most frequently reported AEs recorded for both period 1 and period 2 is provided in Table 6. During period 1, the most commonly reported AEs were gastrointestinal symptoms (diarrhea, anorectal discomfort, abdominal discomfort, abdominal distension). Gastrointestinal symptoms were reported in 25 (76%) patients who received narlaprevir and 4 (50%) patients who received placebo. During period 2, when PEG-IFN-α-2b was added to the treatment regimen, the most commonly

reported AE was influenza-like illness, which was observed in 30 (94%) patients who received narlaprevir and 6 (75%) patients who received placebo. Also during period 2, there was an elevated rate of gastrointestinal selleck inhibitor symptoms. Gastrointestinal-related AEs were reported by 24 (75%) patients who received narlaprevir, compared with no patients in the placebo group. No significant difference in AEs was noted between patients that were treatment-naïve

versus treatment-experienced. Ritonavir coadministration did not significantly affect the AE profile. Three serious AEs (one instance of elevated CRP and two instances of pyrexia) occurred during SOC administration. All three events occurred in the same patient and required hospital admission, but they were not considered related to narlaprevir treatment. No clinically significant changes in blood chemistry or hematological parameters, vital signs, or electrocardiograms occurred in any treatment G protein-coupled receptor kinase group. The present study was the first clinical trial to evaluate narlaprevir in chronic hepatitis C patients and to evaluate a treatment regimen that used a pharmacokinetic enhancer (ritonavir) in combination with an HCV NS3 protease inhibitor for the treatment of hepatitis C. In addition, this was one of the first phase 1b studies to offer treatment with PEG-IFN-α-2b and RBV to all patients following treatment with narlaprevir in order to explore the potential of increasing the RVR and, consequently, the SVR rates. Finally, the first clinical mutational analysis of narlaprevir was performed to investigate the development of NS3/4 genome sequence changes during and after narlaprevir treatment.

[53, 54] In the Medoc and Haut-Medoc regions in France (left margin of the Gironde River), the wines are primarily made of selleck screening library Cabernet Sauvignon grapes (up to 75%), while in the right bank of the Gironde River, due to the arenous soil, the typical wines are primarily made with Merlot grapes as those

from the Pomerol district, which originates the legendary Chateau Petrus.47,50-52 Merlot is a dark blue wine grape used as a blending grape and for varietal wines. The name Merlot is thought to be a diminutive of merle, the French name for the blackbird, probably a reference to the color of the grape. Merlot-based wines usually have medium body with hints of berry, plum, and currant. Its softness and “fleshiness,”

combined with its earlier ripening, makes Merlot a popular grape for blending with the sterner, later ripening Cabernet Sauvignon, which is much higher in tannins.[50] Along with Cabernet Sauvignon, Cabernet Franc, Malbec, and Petit Verdot, Merlot is one of the primary grapes Paclitaxel manufacturer used in Bordeaux wine, and it is the most widely planted grape in the Bordeaux wine regions. Merlot is also one of the most popular red wine varietals in many markets throughout the world.50-54 This flexibility has helped to make it one of the world’s most planted grape varieties. As a varietal wine, Merlot can make soft, velvety wines with plum flavors.[44, 46, 47, 53] Some of the fruit notes commonly associated with Merlot include cassis, black and red cherries, blackberry, blueberry, and plum. Vegetable and earthy notes include black and green olives, nuts, leather, mushrooms, and tobacco. When Merlot has spent significant time in oak (longer than 8 months), the wine may show notes of caramel, chocolate, coconut, coffee bean, smoke, vanilla, and walnut.[46, 47, 53] Wines, especially Avelestat (AZD9668) red

wines, are very different in composition, producing processes and therefore, taste and the ability to please. Consumption habits of different grapes and varietal wines are very peculiar among countries and people. For the good or bad, wine has been linked to headache and migraine attacks as a traditional trigger. Tannins and the phenolic flavonoid components of the red wine, with its ability to interact with the metabolism of certain monoamines as well as its capacity of mobilizing 5-HT, are probably related. However, the methodology of most of the studies discussed in this review and the analysis of the available literature do not allow definitive conclusions regarding the real role of wine in headache. We believe that red wine is indeed a migraine trigger, at least for a percentage of migraineurs, even under regular preventive treatment. In addition, red wines with more tannins are probably worse in triggering migraine attacks. Controlled studies with well-known wines are important to clarify this common belief.

6–3.2 μg/mL. Likewise, DHNA inhibited clinical isolates of H. pylori, resistant to clarithromycin. However, DHNA did not inhibit other Gram negative or anaerobic bacteria in the normal flora of the human intestine. Both H. pylori cellular respiration and adenosine 5′-triphosphate (ATP) generation were dose-dependently inhibited by DHNA. Similarly, the culture filtrates of propionibacterial strains inhibited the growth of H. pylori, and oral administration of DHNA

could eradicate H. pylori in the infected germ-free mice. Conclusions:  The bifidogenic growth stimulator DHNA specifically inhibited the growth of H. pylori including clarithromycin-resistant strains in vitro and its colonization activity in vivo. The bactericidal activity of DHNA was via inhibition of cellular respiration.

These actions of DHNA may have clinical relevance in the eradication of H. pylori. “
“The gastric mucosa of dogs is often DNA Damage inhibitor colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. The colonization of the human gastric mucosa by H. pylori is highly dependent on the recognition of host glycan receptors. Our goal was to define the canine gastric mucosa glycophenotype and to evaluate the capacity of different gastric Helicobacter species to adhere to the canine gastric mucosa. The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa learn more was assessed in cases representative of the canine glycosylation pattern. The canine gastric drug discovery mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. The canine gastric mucosa showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors

may be involved in Helicobacter spp. binding. Helicobacter pylori and NHPH strains differ in their ability to adhere to canine gastric mucosa. Among the NHPH, H. heilmannii s.s. presented the highest adhesion capacity in agreement with its reported colonization of the canine stomach. “
“A combination capsule of bismuth, metronidazole, and tetracycline plus omeprazole given as 10-day therapy has an overall effectiveness of 92–93% in per-protocol analysis (Grade B) with eradication of 86–91% of metronidazole-resistant Helicobacter pylori. This study aimed to explore whether extending the duration to 14 days would improve overall effectiveness per protocol to ≥95% (Grade A) in a population in which metronidazole resistance was anticipated to exist. A one-arm, open-label pilot study of H.

3 mmol/L, mitochondrial degeneration were observed; At 55.5 mmol/L, both the swelling of the endoplasmic reticulum and cytoplasm dissolved occured. Compared with 5.55 mmol/L, the expression of SCF was abundant at Anti-infection Compound Library cost glucose concentrations of 25 and 33.3 mmol/L, scanty at 55.5 mmol/L. Conclusion: Some degree of hyperglucose is in favour of the expression of SCF in SMC. Excessively hyperglucose can damage the SMC ultrastructure. Key Word(s): 1. Glucose; 2. stell cell factor; 3. smoth muscle cell; Presenting Author: ZHU BINHUA Corresponding Author: ZHU BINHUA Affiliations: ying tan people’s hospital Objective: To

investigate the influence of insulin and glucose on the proliferation and the SCF expression of colonic SMC. Methods: SMCs were cultured: at different Insulin concentrations (0, 2.5, 5, 20, 40 and 80 mg/L) for different durations (0, 8, 16 and 24 hours); MTT was used to test the proliferation of SMC; SMCs were cultured at different glucose concentration (5.55, 25, 33.3, 55.5 mmol/L) for 20 days. SMC ultrastructure was observed under the electron microscope, the proliferation of SMC was tested by MTT. Results: The effect of different concentrations Sunitinib nmr of Ins on the proliferation and the SCF expression of colonic SMC: At concentration of 5 mg/L, Ins showed

remarkable role in SMCs proliferation, the same to the concentrations of 20, 40, 80 mg/L; The SMCs incubated at glucose concentrations of 25, 33.3 mmol/L proliferated relative rapidly; At glucose concentrations of 55.5 mmol/L, the SMCs increased slowly. Conclusion: A certain concentration of Ins can promote SMC proliferation; Bacterial neuraminidase Some degree of hyperglucose is in favour of SMC proliferation. Key Word(s): 1. Insulin; 2. glucose; 3. smooth muscle cells; Presenting Author: HUA YANG Additional Authors: BING-QING XIA, BO JIANG Corresponding Author: BO JIANG Affiliations: Department of Gastroenterology, Nanfang Hospital, Southern Medical University; Department

of Gastroenterology, Nanfang Hospital, Southern Medical University Objective: The diagnostic value of stool DNA (sDNA) testing for colorectal neoplasms remains controversial. To compensate for the lack of large-scale unbiased population studies, we performed a meta-analysis to evaluate the diagnostic value of sDNA testing for multiple markers of colorectal cancer (CRC) and advanced adenoma. Methods: he Pubmed, Science Direct, Biosis Review, Cochrane Library, and Embase databases were systematically searched in January 2012 without time restriction. Meta-analysis was performed using a random-effects model using sensitivity, specificity, diagnostic odds ratio (DOR), summary receiver-operating characteristic (sROC) curves, area under the curve (AUC), and 95% confidence intervals (95% CIs) as effect measures.

In our previous study immortalized mouse stellate cell lines that were TLR4 wild type (JS1) and TLR4 knockout (-/-) (JS2) were generated (Guo, et al. Hepatology, 2008). The aim of the present study was to investigate the differential gene expression in these cell lines with or without the stimulation by lipopolysacchirde (LPS), the exogenous TLR4 ligand, and high mobility group box 1 (HMGB1), a potential endogenous TLR4 ligand

and damage pattern molecule that signals the presence of necrosis (Zhang, et al, Lif Sci, 2012). Methods: JS1 LY2157299 clinical trial and JS2 cells that were sub-cultured to 80% confluence were stimulated with normal saline vehicle (control), or 100 ng/ml LPS, or 100 ng/ml HMGB1 for 24 Neratinib nmr hours. The cells were collected with Trizol reagent for RNA extraction. The RNA extracts from the control, LPS and HMGB1 groups were hybridized on a 4644 K Agilent whole mouse genome oligo microarray for the gene expression analysis. Functional analysis of the microarray data was performed using KEGG analyses. Gene interaction network and co-expression network were built on the base of ontology and pathway analysis to which the differentially

expressed genes attributed. Selected genes were validated by real-time polymerase chain reaction (RT-PCR), ELISA and/or Western Blot. Results: The gene expression profiles are different between JS1 and JS2 cells under basal condition and after stimulated with TLR4 ligands. The differentially expressed genes encode extracellular matrix and matrix remodeling proteins,

growth factors and receptors, chemokines and receptors, inflammatory and immune related proteins, as well as transcriptional factors and important signaling molecules. In JS1 cells LPS upregulates genes that belong to the signaling pathways of Toll-like receptors, neurotrophic factor, immune, the spliceosome and nucleotide excision repair and downregulates PPAR signaling, with a variety of MHC molecules, MAPKs, Pik3r3, Prkca, Ikbkb as central regulatory factors. Under HMGB1 stimulation, MAPKs, TRAF6, IGF1R, Gstps appeared to be core regulatory Tangeritin factors in JS1 cells. The gene interaction and co-expression network in JS2 cells under LPS or HMGB1 stimulation are different from JS1 cells, which are simple and lack of core regulatory factors. Conclusion: There were complex gene expression alterations subsequent to the lacking of TLR4 in HSCs. These included key inflammatory, fibrogenic, growth and metabolism related signals in HSCs. These finding emphasizes the complex pathways downstream of TLR4 in this important fibrogenic cell type and the significant consequence of TLR4 signaling on HSC biology and function. Key Word(s): 1. stellate cells; 2. Toll like receptor 4; 3. ligands; 4.

In addition, a patient with mild HA and HR showed the missense p.Glu1704Lys associated with two neutral intronic substitutions

potentially affecting the A3 domain. A case/control study (84/143) permitted estimation of F8 genotype–specific inhibitor risks [OR; prevalence (CI)] in severe-HA patients classifying a high-risk group including multi-exon deletions [3.66; 55% (19–100)], Inv22 [1.8; 24% (19–100)] and nonsense in FVIII-LCh [1.2; 21% (7–59)]; an average risk group including single-exon deletions, indel frameshifts and nonsense-HCh; and a low-risk group represented by missense defects [0.14; 3% (0.6–11)]. Analysis of inhibitor concordance/discordance in related patients indicated additional genetic factors other than F8 genotype for inhibitor formation. No significant EX 527 ic50 inhibitor-predisposing factors related to FVIII

product exposure were found selleck compound in age- and F8 genotype–stratified populations of severe-HA patients. In conclusion, the Argentine HA patient series presents similar global and mutation-specific inhibitor risks than the HA database and other published series. This case-specific information will help in designing fitted therapies and follow-up protocols in Argentina. “
“The multifactorial nature of disability makes it difficult to point out a specific cause for limitations in participation. The conceptual framework of the WHO-ICF (International Classification of Function, Disability and Health) was used to study the determinants participation in patients with severe haemophilia. Outcome was assessed in a single-centre else cohort of 124 patients with severe haemophilia. Joint mobility and muscle strength of the elbows, knees and ankles, in combination with recent X-ray findings (N = 39 only) and the MPQ-DLV pain questionnaire were used to assess Body Functions and Structures. Four performance-based functional tasks and the HAL questionnaire were used to assess Activities. The IPA questionnaire was used to assess Participation. Stepwise and hierarchical regression analysis adjusted for age

and psychological health (Dutch-AIMS 2) was used to associate the various domains of the ICF. Irrespective of age, joint mobility was an important factor in explaining self-reported and performance-based activities. Muscle strength had no significant association with participation. Self-reported activities showed a stronger association with participation than performance-based activities. Adjusted for age and psychological health, joint mobility and pain explained none of the variation in participation. Self-reported activities, however, significantly contributed in explaining participation (25%), whereas performance-based activities (3%) did not. This study adds to the knowledge of determinants of participation in haemophilia.

5 mouse livers, which are comprised of several different liver cell lineages (Supporting Fig. 4A-C).19 These cells, named Hepo-2, Y27632 exhibited low HAI expression (Supporting Fig. 4A,C). Using these Hepo-2 cells we found that interleukin (IL)-β, tumor necrosis factor alpha (TNF-α), and transforming growth factor beta (TGF-β)-1

stimulated HAI-1 expression, whereas TNF-α and HGF marginally induced HAI-2 expression (Fig. 3A). In addition, the expression of an inflammation-related enzyme activated in BA liver,28 COX-2, was significantly elevated in the cells treated with the above factors (Supporting Fig. 4D), and a COX-2 inhibitor, celecoxib, efficiently blocked these stimulatory effects on both HAIs (Fig. 3B). Furthermore, among various bile acids, deoxycholic and lithocholic acids, but not cholic or chenodeoxycholic acid, significantly stimulated HAI-1 expression but not HAI-2 (Fig. 3C). Deoxycholic acid induced the HAI-1 expression in a dose-dependent manner (Fig. 3D). Taken together, these data indicate that selective factors enriched in BA livers activate HAI expression, possibly through the increased expression of COX-2. BMS-777607 supplier To test whether increased HAI-1 or HAI-2 expression plays a role in the fibrosis process in BA livers, portal fibroblasts (PFs) (Fig. 4A) and stellate cells,20 two major cell types responsible for cholestasis-related fibrosis,29, 30 were treated with conditioned media from Hep3B cells stably

Teicoplanin overexpressing HAI-1 or HAI-2 (Fig. 4B) or control media (green fluorescent protein [GFP] or vector) to assay their effects on the fibrogenic activity. Conditioned media containing HAI-1 or HAI-2 significantly increased mRNA levels of collagen I and IV, two common types of collagen expressed in the ductular reaction in BA livers,31 in both cells compared with controls (Fig. 4C). Western blot analysis further confirmed that the conditioned media containing HAI-1 or HAI-2 enhanced the protein levels of collagen I in PFs (Supporting Fig. 5A,B). We also found that conditioned media containing either HAI-1 or HAI-2 significantly increased

PF cell migration, but only HAI-2 significantly increased stellate cell migration (Supporting Fig. 5C). Moreover, colorimetric cell viability (MTT) assays revealed that the HAI-1-rich, and probably HAI-2-rich (from one of two clones) conditioned media, significantly increased the survival of both fibroblasts (Supporting Fig. 5D). Furthermore, recombinant HAI-2 protein (Fig. 4D) significantly up-regulated the expression of Co11a1 and Col4a1 in PFs and the Co11a1 expression in stellate cells. Both HAIs were also expressed in BA livers in cell clusters or single cells with much CK19 and little or no AFP expression (Fig. 2C, arrows; Supporting Fig. 3D), which probably represent a subset of HSCs.15, 24 Thus, we hypothesized that HAI-1 and/or -2 may also have functions in HSCs. To prove this, we examined HAI expression in the developing livers of legally aborted fetuses.

7, 29, 30 It also lends support to the hypotheses that ammonia and inflammatory cytokines may act synergistically31 and that they might induce astrocyte swelling/dysfunction as a common pathogenic endpoint.32 The observation that patients with alcohol-related cirrhosis are more likely to exhibit neuropsychiatric abnormalities than their counterparts with non–alcohol-related cirrhosis is not entirely novel and most likely due to the direct damage that alcohol misuse causes to the brain, regardless Ridaforolimus order of the degree of hepatic involvement.1, 33, 34 In addition, the enhanced activation of the inflammatory cascade observed in patients with alcohol-related cirrhosis

may also play a role. The findings of this study have several direct and indirect implications: First, the discrepancies between EEG and psychometric abnormalities often observed in patients with cirrhosis depend, at least to some extent, on the different pathways leading

to such abnormalities. Second, PHES and EEG analysis are both useful for an optimal HE evaluation in that they reflect different aspects of the pathogenesis of HE and they independently predict the subsequent occurrence of severe overt HE and death. Third, ITF2357 manufacturer for the same reasons, and for purposes of differential diagnosis, the results of a comprehensive neuropsychiatric examination should probably be included in the decision process leading to selection for hepatic transplantation. Finally, it Ergoloid is possible to hypothesize that the effects of ammonia/indole-lowering therapeutic strategies are more likely to be measured by neurophysiological rather than psychometric tools. In contrast, the effects of new drugs aimed at modulating the inflammatory cascade are probably best assessed by psychometry. In conclusion, PHES and EEG abnormalities

in patients with cirrhosis have partially different biochemical correlates and independently predict outcome. If confirmed, these results suggest that, despite the demands of routine hepatology practice for simple tools for the evaluation of neuropsychiatric status, meaningful and prognostically useful results can be obtained only with protocols including both psychometry and neurophysiology and, where possible, measurement of venous ammonia/indole and an inflammatory marker. This will be even more important within research and clinical trial settings. The authors are grateful to Dr. Antonietta Sticca for technical assistance. “
“Cysts in the liver can be classified in several different ways. Congenital cystic disease includes autosomal dominant polycystic kidney disease (ADPKD), simple cysts of the liver, polycystic liver disease (PLD), and the spectrum of diseases that includes autosomal recessive polycystic kidney disease (ARPKD), congenital hepatic fibrosis, and Caroli disease. Acquired cysts include hydatid disease, cystadenoma, and cystadenocarcinoma.

In fact, with the recommended NAS threshold of 5, only 61% of NAFLD cases who died of liver-related causes were diagnosed as having NASH. On the other hand, the Brunt criteria demonstrated the lowest specificity of the four pathologic protocols. In fact, our data indicated that 85% of individuals with NAFLD who did not die from liver-related causes were still diagnosed with NASH by the Brunt criteria, whereas other protocols diagnosed 33% to 60% of those with NASH. However, when patients with Brunt’s

grade 1 NASH were excluded from the NASH group, the proportion of patients with NASH diagnoses among those who did not die from liver-related causes decreased to 41%, and the proportion of correctly included NASH diagnoses (those resulting in LRM) remained at the level of 89%. These data again suggest that Brunt grade 1 for NASH leads to an overdiagnosis of NAFLD BMN 673 ic50 in patients who do not develop progressive liver disease causing liver-related deaths. In an attempt to assess the predictability of NASH diagnoses made by the four separate pathologic protocols, we ran a multivariate analysis. After we controlled for important confounders (age, gender, ethnicity, and presence of obesity and diabetes), a diagnosis of NASH made by the original criteria

for NAFLD subtypes [aHR = 9.94 (95% CI = 1.28-77.08)] and a diagnosis of NASH made by the current study’s criteria for subtypes [aHR = 4.43 (95% CI = 0.97-20.20)] were independent predictors of LRM (Table 4). Again, similarly to the interprotocol agreement analysis, check details we attempted to find the optimal NAS value for predicting LRM. In our analysis, for an association with LRM, the best NAS threshold was 4 (i.e., a patient with NAS ≥ 4 was presumed to have NASH). Nevertheless, the association of this NAS threshold with LRM remained nonsignificant [log-rank P = 0.098, aHR = 2.92 (95% CI = 0.95-8.95)].

Additionally, other thresholds for NAS (both higher and lower) did not return any significant association with LRM. Next, we assessed individual Adenosine triphosphate pathologic features used in the original criteria for NAFLD subtypes for their ability to predict LRM. In a Cox proportional hazards model consisting of all the originally independent pathologic features (i.e., bridging fibrosis and cirrhosis were not included because they could be described as linear combinations of other features) and using each feature as an ordinal parameter, only fibrosis independently predicted LRM. After each pathologic feature was transformed into binary parameters, univariate survival analyses showed that portal inflammation [grade ≥ 2; HR = 6.68 (95% CI = 2.20-20.3), P = 0.0008], ballooning degeneration [grade ≥ 2; HR = 5.32 (95% CI = 1.89-14.9), P = 0.