The ‘high-dose’ Malmö prophylaxis protocol

was aimed to maintain trough FVIII and FIX levels above 1%. Breakthrough joint bleeds were treated with one or more infusions of FVIII or FIX (25–40 IU kg−1) according to severity and until bleeding had stopped. It was recommended that prophylaxis continue life-long. In the Netherlands, prophylaxis was started at an early age according to the individual’s bleeding pattern, generally after the occurrence of at least one or two joint bleeds. The NVP-BEZ235 supplier Dutch regimen involved the administration of 15–25 IU kg−1 of FVIII two or three times a week for haemophilia A cases, and 30–50 IU kg−1 of FIX once or twice a week for haemophilia B cases. The intensity of prophylaxis was adjusted based on spontaneous breakthrough bleeding into joints and not increased according to the subject’s body weight alone. Trough levels of FVIII or FIX were not taken into consideration when adjusting prophylactic treatment. selleck products It was recommended that prophylaxis continue throughout adulthood. The Canadian dose-escalation primary prophylaxis study was started in 1997. In this single arm, prospective study, boys’ ages 1 year to 30 months with severe haemophilia A, no evidence of a circulating inhibitor to FVIII and absence of any overt joint disease were

started on once weekly infusions of FVIII (50 IU kg−1). If clinically significant bleeding into muscles and/or joints occurred, the frequency of FVIII infusions was increased to twice weekly (dose 30 IU kg−1); continuation of bleeding resulted in escalation of the prophylaxis regimen 上海皓元 to 25 IU kg−1 on alternate days. Criteria for escalation included: ≥ 3 clinically determined bleeds into any one joint over a consecutive 3-month period; ≥ 4 significant soft tissue/joint bleeds over a consecutive 3-month period and ≥ 5 bleeds into any one joint while on the same dosage (step) of factor therapy over any period of time. The interim results of this study have been reported [16], and 10-year follow-up results were presented at the 2009 International

Society on Thrombosis and Hemostasis Congress [22]. The Canadian primary prophylaxis study is now closed to patient accrual, but follow-up of enrolled cases is ongoing. Key results from these three long-term prophylaxis studies are as follows: 1 Compared to on-demand therapy, intermediate-dose prophylaxis (the Dutch protocol) started at an early age in boys with severe haemophilia results in significantly fewer joint bleeds, a better joint status and a more favourable health-related quality of life [20]. The results of the retrospective Swedish and Dutch cohort studies continue to be debated. At the centre of this debate is the issue of when should primary prophylaxis be started in boys with haemophilia A? Data from the Swedish and Dutch studies suggest that primary prophylaxis should be started at an early age but can be individualized based on the bleeding pattern in the individual child [23,24].

The ‘high-dose’ Malmö prophylaxis protocol

was aimed to maintain trough FVIII and FIX levels above 1%. Breakthrough joint bleeds were treated with one or more infusions of FVIII or FIX (25–40 IU kg−1) according to severity and until bleeding had stopped. It was recommended that prophylaxis continue life-long. In the Netherlands, prophylaxis was started at an early age according to the individual’s bleeding pattern, generally after the occurrence of at least one or two joint bleeds. The Midostaurin concentration Dutch regimen involved the administration of 15–25 IU kg−1 of FVIII two or three times a week for haemophilia A cases, and 30–50 IU kg−1 of FIX once or twice a week for haemophilia B cases. The intensity of prophylaxis was adjusted based on spontaneous breakthrough bleeding into joints and not increased according to the subject’s body weight alone. Trough levels of FVIII or FIX were not taken into consideration when adjusting prophylactic treatment. MS 275 It was recommended that prophylaxis continue throughout adulthood. The Canadian dose-escalation primary prophylaxis study was started in 1997. In this single arm, prospective study, boys’ ages 1 year to 30 months with severe haemophilia A, no evidence of a circulating inhibitor to FVIII and absence of any overt joint disease were

started on once weekly infusions of FVIII (50 IU kg−1). If clinically significant bleeding into muscles and/or joints occurred, the frequency of FVIII infusions was increased to twice weekly (dose 30 IU kg−1); continuation of bleeding resulted in escalation of the prophylaxis regimen medchemexpress to 25 IU kg−1 on alternate days. Criteria for escalation included: ≥ 3 clinically determined bleeds into any one joint over a consecutive 3-month period; ≥ 4 significant soft tissue/joint bleeds over a consecutive 3-month period and ≥ 5 bleeds into any one joint while on the same dosage (step) of factor therapy over any period of time. The interim results of this study have been reported [16], and 10-year follow-up results were presented at the 2009 International

Society on Thrombosis and Hemostasis Congress [22]. The Canadian primary prophylaxis study is now closed to patient accrual, but follow-up of enrolled cases is ongoing. Key results from these three long-term prophylaxis studies are as follows: 1 Compared to on-demand therapy, intermediate-dose prophylaxis (the Dutch protocol) started at an early age in boys with severe haemophilia results in significantly fewer joint bleeds, a better joint status and a more favourable health-related quality of life [20]. The results of the retrospective Swedish and Dutch cohort studies continue to be debated. At the centre of this debate is the issue of when should primary prophylaxis be started in boys with haemophilia A? Data from the Swedish and Dutch studies suggest that primary prophylaxis should be started at an early age but can be individualized based on the bleeding pattern in the individual child [23,24].

MK-6096 for insomnia, painful diabetic neuropathy, depression, and migraine. A Phase 2 study entitled “A Study of the Safety and Efficacy of MK-6096 for Migraine Prophylaxis in Participants With Episodic Migraine” (NCT01513291) was initiated in early 2012, with an expected enrollment of 450 subjects. The results from this proof of concept study were expected in 2013 but have not been announced. However, a company drug development pipeline update in April 2013 indicated that filorexant was now being developed for only insomnia, suggesting

a discontinuation Ibrutinib ic50 of its development for migraine prevention. BGG is an AMPA antagonist and putative anticonvulsant being developed by Novartis. A Phase 2 acute migraine study using single doses of the drug was completed in late 2010 but the data were never published. In addition, another Phase 2 study entitled “A Randomized, Double Blind, Placebo Controlled Study to Assess Efficacy, Safety and Tolerability

of BGG492 in Migraine Prevention” (NCT01617941) was initially planned to begin in 2012. However, enrollment into this planned 90 subject study was suspended in January 2013. BGG492 is also in clinical development for epilepsy and tinnitus. Crizotinib A small (n = 70 subjects) Phase 3 study entitled “Efficacy and Safety of Cyclobenzaprine Hydrochloride Extended Release for the Treatment of Chronic Migraine” (NCT01151787) began in 2010. The study drug (cyclobenzaprine)

is an extended release formulation of the marketed product Flexeril®. The principle outcome variable will be the mean total number of migraine/migrainous headache days, which will be calculated for the month prior to enrollment in the study (pretreatment) and then calculated for the third month after study treatment 上海皓元 (posttest) after taking 15 mg of cyclobenzaprine or placebo. The study is being conducted at a single site in the United States (ie, The Headache Center at Kennedy Health Alliance in New Jersey). The study is scheduled to complete in 2014. Trigemina, Inc., is developing TI-001, an intranasal formulation of the hormone oxytocin. The results of a single-dose, placebo-controlled, double-blind study found that TI-001was safe and effective in the acute treatment of chronic migraine. The data from this study included 40 subjects with chronic migraine who received 32 units of nasally applied oxytocin dose of the agent and were asked to rate their pain, nausea, photophobia, and phonophobia on a 4-point scale (indicating severe, moderate, mild, or none) after dosing with TI-001. At 2 hours after dosing, nasal OT reduced pain by 2 categories in 42% of subjects compared with 11% for placebo. Photophobia and phonophobia were also decreased compared to placebo. The authors concluded that nasal oxytocin may be a viable alternative for the treatment of chronic migraine headache.

5 hours after Jo2 administration (Fig. 5C). At 5 hours after Jo2 administration, marked phosphorylation and subsequent degradation of BimEL and reduction of the cytochrome c level in the mitochondrial fraction were seen in WT mice, whereas these changes were significantly suppressed in ASK1−/− mice (Fig. 5D). As reported,19 administration of a JNK inhibitor reduced Jo2-induced BimEL phosphorylation and serum ALT elevation. However, administration of a p38 inhibitor had no detectable effect on BimEL phosphorylation

or liver injury (Fig. 5E,F). These results suggest that ASK1 plays an important role in Fas-induced activation of the JNK–Bim–mitochondrial apoptotic pathway. Next, Selleckchem Regorafenib to examine whether ASK1 may be involved in a Fas-induced mitochondria-independent apoptotic pathway, we used primary thymocytes, which are independent of mitochondria for Fas-induced apoptosis (so-called type I cells). Fas-induced activation of JNK and p38 was reduced in ASK1−/− thymocytes, whereas caspase-3 activation and cell viability were comparable between WT and ASK1−/− thymocytes (Supporting Fig. 1A,B), suggesting that ASK1 is not required for the mitochondria-independent

apoptotic pathway. Recently, Fas signaling was reported to play a role in not only cancer cell apoptosis, but also cancer cell proliferation.26 JNK has also been shown to be one of the main mediators of Fas-mediated proliferative Bortezomib signals. To investigate whether ASK1 participated in Fas-mediated hepatocyte proliferation, we injected Jo2 to WT and ASK1−/− mice after partial hepatectomy, which is known to convert

Fas signaling from MCE公司 apoptotic to proliferative.27 As reported,26, 27 Jo2 injection after partial hepatectomy induced JNK phosphorylation and accelerated hepatocyte proliferation without liver injury (Supporting Fig. 2A,B). Although liver regeneration after partial hepatectomy and Jo2-induced JNK phosphorylation were slightly impaired in ASK1−/− mice (especially the upper band corresponding to JNK2), there was no significant difference in Jo2-mediated acceleration of hepatocyte proliferation (Supporting Fig. 2A,B). Thus, ASK1 seemed to regulate the apoptotic, but not proliferative, function of JNK in Fas signaling. To further confirm the involvement of ASK1 in Fas-induced hepatocyte apoptosis, we examined whether the reintroduction of ASK1 to ASK1−/− mouse liver restored sensitivity to Fas. We injected an adenoviral vector encoding either Ad-ASK1 or LacZ into the tail vein of ASK1−/− mice. ASK1 protein was successfully expressed in ASK1−/− mouse liver, as much as that in WT mouse liver, at 48 hours after Ad-ASK1 injection (Fig. 6A). Immunohistochemical analysis using anti-HA antibody revealed that ≈70%-80% of hepatocytes were transduced with the ASK1 gene (Fig. 6B). The reintroduction of ASK1 did not affect the serum ALT level or liver histology.

The lack of significance between prey with high and low levels of unpalatability may indicate that low levels of unpalatability have an intermediate effect on predation. Indeed, it may be profitable TSA HDAC order for aposematic prey to invest in lower levels of unpalatability in light of the metabolic costs of chemical defences (Nishida, 2002; Mappes et al., 2005). However, the

lack of significance in predation rates between low unpalatability prey and cryptic prey suggests that there is a benefit to being more unpalatable, particularly as predators may strategically consume aposematic prey based on factors such as hunger and toxin load (Sherratt, Speed & Ruxton, 2004; Skelhorn & Rowe, 2007). The lack of significance between the two types of unpalatable prey could also have been caused by predators moving between sites, because in two of the sites, the colour treatments were reversed and this may have confused predator learning. However, we consider this unlikely because both conspicuous prey types possessed some level of chemical defence, PLX4032 supplier and the colour treatments were never reversed within a single site. The differences between complete and partial consumption of cryptic and aposematic baits are readily explained by ‘go-slow’ predation (Guilford, 1994), a strategy in which predators cautiously sample aposematic prey and

reject those that are unpalatable without necessarily killing them. This allows predators to avoid the cost of consuming chemically defended prey, while still being able to sample novel or rare conspicuous species; at the same time, aposematic individuals may avoid the disproportionately high mortality rates that are often a consequence of conspicuousness. Go-slow predation may therefore represent a potential defensive advantage of aposematism over crypsis, especially 上海皓元 because aposematic insects can survive sampling and rejection by both captive and wild avian predators (Wiklund & Jarvi, 1982; Sillen-Tullberg, 1985; Pinheiro, 1996). Go-slow predation

can also help to explain the evolution and spread of novel aposematic species, which has been traditionally considered problematic because of the presence of anti-apostatic (positive frequency dependent) selection (Endler, 1988; Skelhorn & Ruxton, 2007), by providing a benefit for honest signalling (Holen & Svennugsen, 2012). To date, several experiments with captive avian predators have demonstrated the presence of go-slow predation in response to novel aposematic prey (Sillen-Tullberg, 1985; Gamberale-Stille & Guilford, 2004; Skelhorn & Rowe, 2006a,b; Halpin et al., 2008), but as far as we are aware, it has not yet been documented in wild predators. It is important to note that our results could also have been caused by simple taste-rejection behaviour. Taste rejection differs from go-slow predation in that predators reject prey based solely based on palatability, and do not exhibit cautious attack behaviour when chemically defended prey are conspicuous.

“
“(Headache

2010;50:256-263) Aim.— To estimate the proportion of individuals with migraine using triptan therapy as a function of their cardiovascular (CV) Selleck LGK-974 profile and disease severity. Methods.— As a part of the American Migraine Prevalence and Prevention study, we identified migraineurs representative of the U.S. adult population. Triptan use was estimated as a function of presence of CV disease (CVD), of CV risk factors, and by level of migraine-related disability. Results.— Our sample consists of 6102 individuals with migraine. Compared with migraineurs without risk factors for CVD, triptans were significantly less likely to be used in individuals with diabetes (11.5% vs 18.3%, OR = 0.6, 95% CI = 0.5-0.7), hypertension (14.8%, OR = 0.8, 0.7-0.9) and by smokers (12.9%, OR = 0.7, 0.6-0.8). Similar findings were seen for individuals with established CVD. As contrasted to individuals without CVD, those with myocardial infarct (8.5% vs 18.5%, OR = 0.4, 0.3-0.7), stroke (7%, OR = 0.6, 0.3-0.9) and heart surgery (9.3%, OR = 0.5, 0.4-0.7) were less likely to use triptans. Use of triptan increased as a function of disability regardless of CVD status or presence of CV risk factors. Conclusion.— Triptan use is lower in those with vs without CV risk, suggesting that doctors and/or patients fear using triptans

in individuals at risk to CVD. Furthermore, triptan use in those with established CVD increases with headache-related disability, suggesting that patients and providers balance risks and selleckchem MCE公司 benefits. Additional and analytical data are needed on the safety of triptans in the setting of CVD risk. This study has not assessed adequacy of care. “
“Objectives.— To determine the 1-year prevalence of headache and clinical characteristics of primary headaches among school children in South Korea. Background.— Many population-based studies have estimated the 1-year prevalence of headache, migraine, and tension-type headache (TTH).

The results of those studies vary in terms of race and region. There have been few epidemiological population-based studies of headache in children and adolescents in Korea. Methods.— We conducted a cross-sectional school-based study of a randomized and proportional sample of 5360 boys and girls. All 180 sampled schools participated in this study. The questionnaires collected demographic data in addition to specific questions about headache according to the International Classification of Headache Disorder criteria, 2nd Edition. Valid questionnaires were returned by 94.1% of the sample population. Modified criteria changed the “duration” of migraine (>1 hour instead of 4 hours). Results.— The prevalence of headache among school children was 29.1% (1465/5039) in South Korea. The prevalence of headache in girls (33.4%) was significantly higher than in boys (24.4%) (P < .001). The mean age of students with headaches (14.02 ± 3.

“
“(Headache

2010;50:256-263) Aim.— To estimate the proportion of individuals with migraine using triptan therapy as a function of their cardiovascular (CV) http://www.selleckchem.com/products/i-bet-762.html profile and disease severity. Methods.— As a part of the American Migraine Prevalence and Prevention study, we identified migraineurs representative of the U.S. adult population. Triptan use was estimated as a function of presence of CV disease (CVD), of CV risk factors, and by level of migraine-related disability. Results.— Our sample consists of 6102 individuals with migraine. Compared with migraineurs without risk factors for CVD, triptans were significantly less likely to be used in individuals with diabetes (11.5% vs 18.3%, OR = 0.6, 95% CI = 0.5-0.7), hypertension (14.8%, OR = 0.8, 0.7-0.9) and by smokers (12.9%, OR = 0.7, 0.6-0.8). Similar findings were seen for individuals with established CVD. As contrasted to individuals without CVD, those with myocardial infarct (8.5% vs 18.5%, OR = 0.4, 0.3-0.7), stroke (7%, OR = 0.6, 0.3-0.9) and heart surgery (9.3%, OR = 0.5, 0.4-0.7) were less likely to use triptans. Use of triptan increased as a function of disability regardless of CVD status or presence of CV risk factors. Conclusion.— Triptan use is lower in those with vs without CV risk, suggesting that doctors and/or patients fear using triptans

in individuals at risk to CVD. Furthermore, triptan use in those with established CVD increases with headache-related disability, suggesting that patients and providers balance risks and R788 supplier 上海皓元医药股份有限公司 benefits. Additional and analytical data are needed on the safety of triptans in the setting of CVD risk. This study has not assessed adequacy of care. “
“Objectives.— To determine the 1-year prevalence of headache and clinical characteristics of primary headaches among school children in South Korea. Background.— Many population-based studies have estimated the 1-year prevalence of headache, migraine, and tension-type headache (TTH).

The results of those studies vary in terms of race and region. There have been few epidemiological population-based studies of headache in children and adolescents in Korea. Methods.— We conducted a cross-sectional school-based study of a randomized and proportional sample of 5360 boys and girls. All 180 sampled schools participated in this study. The questionnaires collected demographic data in addition to specific questions about headache according to the International Classification of Headache Disorder criteria, 2nd Edition. Valid questionnaires were returned by 94.1% of the sample population. Modified criteria changed the “duration” of migraine (>1 hour instead of 4 hours). Results.— The prevalence of headache among school children was 29.1% (1465/5039) in South Korea. The prevalence of headache in girls (33.4%) was significantly higher than in boys (24.4%) (P < .001). The mean age of students with headaches (14.02 ± 3.

The results show that high dietary fat supplied as cocoa butter (a fat source relatively high in saturated fat) and excess cholesterol

interacted to produce both the metabolic and hepatic features of NASH, whereas neither dietary factor alone was sufficient to cause significant disease. These studies provide clear evidence that the hepatic and metabolic effects induced by combined high dietary fat and cholesterol were substantially greater than the sum of the separate effects of each dietary component LY2606368 order alone. How does high dietary cholesterol synergize with high dietary fat to cause NASH? One explanation for this synergy may be the activation of the liver X receptor (LXR) pathway by cholesterol (or more specifically,

by the oxysterol metabolites of cholesterol) resulting in a gene expression program designed to detoxify and eliminate cholesterol.3, MK0683 4 The membrane disruptive effects of mildly amphipathic free cholesterol can be prevented by cholesterol esterification with fatty acids to form highly lipophilic cholesterol esters. Perhaps in an effort to preserve fatty acid availability for cholesterol esterification, LXR activation also inhibits mitochondrial β-oxidation (Fig. 1).5 This is of little consequence when the liver is not faced with a surfeit of fatty acids. However, when the liver must deal with MCE excessive fatty acids, either from de novo lipogenesis, as can occur with fructose feeding, or from excess dietary fat with spillover of fatty acids into the circulation from lipoprotein lipase-mediated hydrolysis or adipose insulin resistance, the stage is set for lipotoxic liver injury. The inability of the liver to handle fatty acids through oxidative pathways or the formation of triglyceride may predispose to the excessive formation of other fatty acid derivatives such as diacylglycerols, ceramides, and lysophosphatidyl choline that have

been proposed to cause lipotoxic liver injury manifesting itself as NASH.6 Another important observation by Savard et al. in their study of the combined effects of a high cholesterol, high-fat diet was that the mice fed this combination exhibited a much greater increase in weight than mice fed a control diet or even the high-fat diet alone. This was despite having the same daily caloric intake as the control mice and a lower daily caloric intake than the mice fed the high-fat diet. One explanation is that excess dietary cholesterol facilitates fat absorption. Increased fat absorption associated with a high cholesterol diet is a known phenomenon7 and in fact the combined diet fed mice did exhibit slightly less fecal fat loss, indicating greater absorption.

GFP fluorescence was collected through a 530/40

nm bandpass filter. Cells were sorted at a rate of 30,000 events per second using a sort purify 1 drop window. GFP-positive cells were sorted into Waymouth’s this website culture medium supplemented with 10% FBS and plated. X-gal staining was performed in the same manner as described above. Liver tissues were fixed in 10% buffered formalin, embedded in paraffin, and sectioned at 5 μm thickness. Sections were stained with Sirius red solution (0.1% Direct Red 80 in saturated picric acid) to visualize collagen deposition. Hepatocytes were isolated and cultured from triple transgenic mice ROSA26 stop β-gal, Alb Cre, and Coll GFP. Hepatocytes were GFP-negative before TGFβ-1 treatment or after 48 hours culture without TGFβ-1 (Fig. 1A, left and middle). Treatment of hepatocytes with 3 ng/mL TGFβ-1 for 48 hours induced a fibroblast-like morphological change (Fig. 1A, upper APO866 cell line right), consistent with previous findings.6, 13 TGFβ-1 treatment not only induced a fibroblast-like morphological change, but also activated the collagen α1(I) promoter, as demonstrated by GFP expression in TGFβ-1-treated hepatocytes (Fig. 1A, middle right). The messenger RNA (mRNA) level of collagen α1(I) in TGFβ-1-treated hepatocytes was increased to a level comparable to culture-activated HSCs (Supporting Fig. S1A). The increase was not blocked by gliotoxin, which eliminates potentially

contaminating HSCs, as demonstrated by inhibition of the level of desmin, a marker for HSCs (Supporting Fig. S1B). β-Gal expression of

GFP-positive cells indicated that these cells were not contaminating nonparenchymal cells, but instead originally derived from hepatocytes (Fig. 1A, bottom, right). This result was confirmed by another experiment in which X-gal staining was followed by immunocytochemistry for GFP. Consistently, hepatocytes doubly positive for GFP and β-gal appeared upon stimulation with TGFβ-1 (Fig. 1B, right). A few GFP-positive cells were seen even in the absence of TGFβ-1 (Fig. 1B, middle). However, they were never positive for β-gal, indicating they were contaminating nonparenchymal cells. Liver fibrosis was induced in triple transgenic mice ROSA26 stop β-gal, Alb Cre, and Coll GFP by eight injections with CCl4 (Supporting medchemexpress Fig. S2). GFP-positive cells (collagen-expressing cells) were seen along fibrotic septa (Fig. 2, upper). However, they were never positive for β-gal, as demonstrated by the lack of double-positive cells in merged images of GFP and X-gal staining (Fig. 2, bottom). Higher-magnification images clearly show that the GFP-positive cells are present exclusively in the X-gal-negative area (Supporting Fig. S3). The absence of hepatocyte-derived collagen-expressing cells (i.e., cells double positive for GFP and β-gal) was confirmed by cell isolation; double-positive cells were not seen in the whole liver cell fraction (Fig.

GFP fluorescence was collected through a 530/40

nm bandpass filter. Cells were sorted at a rate of 30,000 events per second using a sort purify 1 drop window. GFP-positive cells were sorted into Waymouth’s Cobimetinib solubility dmso culture medium supplemented with 10% FBS and plated. X-gal staining was performed in the same manner as described above. Liver tissues were fixed in 10% buffered formalin, embedded in paraffin, and sectioned at 5 μm thickness. Sections were stained with Sirius red solution (0.1% Direct Red 80 in saturated picric acid) to visualize collagen deposition. Hepatocytes were isolated and cultured from triple transgenic mice ROSA26 stop β-gal, Alb Cre, and Coll GFP. Hepatocytes were GFP-negative before TGFβ-1 treatment or after 48 hours culture without TGFβ-1 (Fig. 1A, left and middle). Treatment of hepatocytes with 3 ng/mL TGFβ-1 for 48 hours induced a fibroblast-like morphological change (Fig. 1A, upper Y-27632 manufacturer right), consistent with previous findings.6, 13 TGFβ-1 treatment not only induced a fibroblast-like morphological change, but also activated the collagen α1(I) promoter, as demonstrated by GFP expression in TGFβ-1-treated hepatocytes (Fig. 1A, middle right). The messenger RNA (mRNA) level of collagen α1(I) in TGFβ-1-treated hepatocytes was increased to a level comparable to culture-activated HSCs (Supporting Fig. S1A). The increase was not blocked by gliotoxin, which eliminates potentially

contaminating HSCs, as demonstrated by inhibition of the level of desmin, a marker for HSCs (Supporting Fig. S1B). β-Gal expression of

GFP-positive cells indicated that these cells were not contaminating nonparenchymal cells, but instead originally derived from hepatocytes (Fig. 1A, bottom, right). This result was confirmed by another experiment in which X-gal staining was followed by immunocytochemistry for GFP. Consistently, hepatocytes doubly positive for GFP and β-gal appeared upon stimulation with TGFβ-1 (Fig. 1B, right). A few GFP-positive cells were seen even in the absence of TGFβ-1 (Fig. 1B, middle). However, they were never positive for β-gal, indicating they were contaminating nonparenchymal cells. Liver fibrosis was induced in triple transgenic mice ROSA26 stop β-gal, Alb Cre, and Coll GFP by eight injections with CCl4 (Supporting 上海皓元医药股份有限公司 Fig. S2). GFP-positive cells (collagen-expressing cells) were seen along fibrotic septa (Fig. 2, upper). However, they were never positive for β-gal, as demonstrated by the lack of double-positive cells in merged images of GFP and X-gal staining (Fig. 2, bottom). Higher-magnification images clearly show that the GFP-positive cells are present exclusively in the X-gal-negative area (Supporting Fig. S3). The absence of hepatocyte-derived collagen-expressing cells (i.e., cells double positive for GFP and β-gal) was confirmed by cell isolation; double-positive cells were not seen in the whole liver cell fraction (Fig.