Hwang et al. reported that the total amount of hepatic steatosis in nine living donors changed significantly from 48.9% ± 25.6% before 2–6 months of weight reduction (loss of 5.9% ± 2.0% of initial body weight) to 20.0% ± 16.2% after

weight reduction.40 All donors recovered uneventfully and all recipients survived AUY-922 more than 15 months. This type of intervention, if the general condition of the recipient permits, will contribute to expanding the pool of marginal living doors. Hepatitis C virus (HCV) recurrence is universal and is associated with poor graft and patient survival. Pre-emptive antiviral therapy started within weeks of transplantation is limited by tolerability. Rates of sustained virological response (SVR) vary from 8% to 39%.41,42 Post-transplant antiviral therapy initiated upon histological evidence of recurrence is therefore the mainstay of treatment. Most recently, KU57788 the Kyoto group reported that a combination of pegylated interferon alpha-2b and ribavirin achieved a 50% SVR rate for recurrent hepatitis C genotype 1b.43 In contrast, pretransplant therapy is a favorable option for well-compensated patients with HCC or

mildly decompensated liver cirrhosis, since up to two-thirds of patients who become HCV RNA-negative on treatment are suggested to be HCV infection-free after LT.42 From the analysis of 275 patients who underwent pretransplant antiviral therapy selleck inhibitor with mostly mild to moderate liver decompensation, receiving either LDLT or DDLT, rates of on-treatment SVR were 30% (range, 18–56%) in genotype

1 and 83% (range, 82–100%) in genotype 2/3 recipients.42,44 Whether HCV-infected LDLT recipients show worse graft outcomes than HCV-infected DDLT recipients has been controversial. The retrospective A2ALL cohort study recently demonstrated that graft survival did not differ significantly for recipients of LDLT compared with DDLT once centers have sufficient experience with LDLT.45 The risks to the healthy live donor represent the greatest disadvantage for LDLT. As noted earlier, the healthy live donor undergoes major surgery for no direct, physical benefit. The Japanese Liver Transplantation Society collected data from 3565 live donors and reported that 299 donors (8.4%) suffered complications related to live donation, with one donor death.46 A worldwide systematic review reported that donor morbidity ranged from 0% to 100%, with a median of 16.1%.47 Biliary complications and infections were the most commonly reported donor morbidities, with median frequencies of 6.2% and 5.8%, respectively. Based on the estimate of 14 000 LDLTs performed worldwide, the donor death rate is 0.1–0.3%.1 The A2ALL group investigated the rate and severity of complications in 393 living donors.48 Eighty-two donors (21%) experienced one complication, and 66 (21%) had two or more.

1-IDO cells. Murine model of gastric cancer was established. Tumor-bearing mice was divided into 6 gmups: MFC non-transfected group (Group A), pcDNA3.1 group (Group

B), pcDNA3.1-IDO group (Group C), 1-MT-pcDNA3.1-IDO treatment group (Group D), FOLFOX4-pcDNA3.1-IDO treatment group (Group E) and FOLFOX4+1-MT-pcDNA3.1-IDO treatment group (Group F). We can observe the mice tumor cases, differences of tumor weight, and then detect the expression of IDO PD0325901 supplier in tumor tissues by immunohistochemical. The method of RT-PCR was used to detect the expression of IDOmRNA of the spleen organization of tumor-bearing mice. The method of Flow cytometry was used to detect the expression of CD11c, and CD86, and CD80, and Major histocompatibility Antigen complex II (MHC-II) in the surface of Dendritic Cells of the spleen of tumor-bearing mice. Results: Compared learn more with Group A and Group B, the tumor

on Group C grows faster, the weight of tumor increases significantly (P < 0.05). Compared with Group C, the weight of tumor on Group D, Group E and Group F decreases significantly (P < 0.05). The tumor inhibitory rates were 8.91%, 80.20%, 86.13% respectively. There were significant differences between them (P < 0.05), Compared with Group D and Group E, the weight of tumor on Group F decreases significantly (P < 0.05). IDO expression in tumor tissues: Treatment group, cancer cells were less color, especially in the combination group there was even no brown color cells; the gastric selleck kinase inhibitor cancer cells in control group were more, which were arranged in sheets, much larger, colored extensively, and more colored granules. Compared with Group A and Group

B, the expression of IDOmRNA of Group C increases significantly (P < 0.05). The expression of IDOmRNA of Group A and Group B are not significantly different (P > 0.05). Compared with the Group C, the expression of IDOmRNA of group D, group E and group F reduces significantly (P < 0.05). Compared with Group F, the expression of IDOmRNA of Group D and Group E increases significantly (P < 0.05). Compared with the Group C, the number of DC in the spleen of Group A, Group B and Group D increases Slightly. Compared with the Group C, the number of DC in the spleen of Group D and GroupE increases significantly (P < 0.05). Compared with the Group F, the number of DC in the spleen of Group D and Group E reduces Slightly. Compared with the Group C, the surface molecule including CD86, CD80 and MHC-II of DC of the spleen of Group A, Group B, Group D, Group E and Group F increases significantly (P < 0.05). Compared with the Group F, the surface molecule including CD86, CD80 and MHC-II of DC of the spleen of Group D and Group E reduces significantly (P < 0.05).

General effects such as steric hindrance or changes in electrostatic binding properties of the modified rFVIIa to its receptors are probably responsible for this impairment rather than a loss of specific recognition of the receptors, which could explain FK228 molecular weight near normal activation of factor X by glycoPEGylated rFVIIa on TF expressing cells while its uptake is reduced. “
“Summary.  Inpatient costs comprise >50% of annual healthcare costs for haemophilia patients with inhibitors but no reports exist on inpatient

resource use and costs at a US national level. To quantify inpatient resource use and costs for on-demand treatment of bleeds of US haemophilia patients with inhibitors and compare costs and treatment duration between Factor VIII bypassing agents (BAs). Stays with haemophilia A from 2003–2008 were identified from inpatient billing records. Presence of inhibitors was inferred through use of BA; recombinant activated Factor VII and plasma-derived activated prothrombin complex concentrate. Duration and number of infusions of BA, length of stay, use of opioid-containing analgesics and

costs were assessed and compared. Among 1322 stays mean BA treatment duration was 4.6 days with 4.9 infusions, 6.1 nights spent in hospital, and 58% administered opioid-containing analgesics. In unadjusted JQ1 cell line analyses there were significant differences in the above mentioned outcomes by BA use, reflecting underlying differences between the two patient populations. Average inpatient costs were $82 911. In adjusted analyses, African-American race, greater disease severity, hospital region outside the southern US and older age (cost model only) were significant predictors of longer BA treatment duration and higher costs. The economic burden of inpatient find more on-demand treatment of haemophilia with inhibitors is substantial and is associated with lengthy stays, high costs and inadequate pain relief. Availability of more effective BAs could reduce the need for re-treatment, reducing

treatment costs and other medical costs, while improving health related quality of life. “
“Summary.  Muscle haematomas (MH) represent 10–25% of all bleeds in patients with severe haemophilia. We performed a cross-sectional survey on current practice in the management of MH with participation from 22 consultants. The respondents reported 492 MH/year, corresponding an average of 25/centre, mostly associated with trauma. Iliopsoas (55%), calf (18%) and thigh (18%) bleeds were scored as most serious. Half of the respondents distinguished between contusion and strains, whereas the majority (68.2%) did not categorize bleedings as intra- or intermuscular, although 77.3% routinely used ultrasound. Half of the respondents used a standard protocol for the management of MH. Twenty of 22 (90.

Daily rhythms are controlled by a circadian clock, entrained to the overriding cue of light intensity (a ‘zeitgeber’ in the terms of Lorenz & Kickert, 1981), and in evolutionary terms, responding to a zeitgeber facilitates efficient use of the environment (Kronfeld-Schor et al., 2001). Here, it triggers appropriately timed, physiological and behavioural responses (Heldmaier et al., 1989; Refinetti, Nelson & Menaker, 1992; Aronson et al., 1993), and facilitates interspecific coexistence (Schoener, 1974; Richards, 2002). Though temporal partitioning in communities has

never been a strong focus of ecology (Kronfeld-Schor Wnt inhibitor & Dayan, 2003) and biologists are aware that there is a degree of rigidity in the response to light, there are few field data to reveal the plasticity of this endogenous rhythmicity.

In particular, little is known of what triggers are likely to mask the zeitgeber, although there are examples where one species causes another to adopt an opposite activity pattern [e.g. mink Neovison vison : otter Lutra lutra and fox Vulpes vulpes : rat Rattus norvegicus interactions (Fenn & Macdonald, 1995; Harrington et al., 2009)]. Furthermore, in the context of landscapes increasingly dominated by people, behavioural plasticity may reduce the threats to a species but will incur a cost [e.g. hyaenas Crocuta crocuta (Boydston et al., 2003)]. With the African wild dog or painted hunting dog (Courchamp, Opaganib in vivo Rasmussen & Macdonald, 2002) Lycaon

pictus (hereinafter referred to as Lycaon) representing a monotypic genus and listed as endangered by the International Union for Conservation of Nature/Species Survival Commission (Woodroffe, Ginsberg & Macdonald, 1997), the aim of this article is therefore to explore (1) the relationship between activity patterns of Lycaon and sympatric competition under ‘natural’ see more conditions of coexistence; (2) plasticity in response to high anthropogenic activity; (3) potential costs of sub-optimization and masking behaviours. We present data from two parapatric Lycaon populations in Zimbabwe, and their competitors. As circadian entrainment is essentially light driven, we make our measurements relative to solar and lunar light cues. Lycaon are eusocial (Sherman et al., 1994; Rasmussen et al., 2008) kin-selected, obligate cooperative breeding canids (Courchamp et al., 2002), living in packs of up to 20 adults. Usually, only the alpha pair breeds, with the remaining adults being reproductively suppressed. It is among the most endangered large carnivores in Africa, with most of the remaining packs being in populations too small to be viable (Woodroffe et al.

The authors

thank the Cellular and Molecular Morphology Core of the Texas Medical Center Digestive Diseases Center (NIDDK-P30-DK056338) and Pamela Parsons for help with immunohistochemistry, the Clinical Pathology Laboratory of Texas Children Hospital for liver function tests, and Dr. Juan Marini (BCM) for help with submandibular bleeding. Additional Supporting Information may be found in the online version of this article. “
“Interleukin (IL)-20 is a proinflammatory cytokine of the IL-10 family and involved in rheumatoid arthritis, atherosclerosis, stroke, and osteoporosis. However, the pathophysiological Ceritinib roles of IL-20 in liver injury have not been extensively studied. We explored the involvement of IL-20 in liver injury and the therapeutic potential of IL-20 antagonists for treating liver fibrosis. Compared with normal liver tissue from healthy individuals, the amount of IL-20 was much higher in hepatocytes and hepatic stellate cells in liver biopsies from patients with fibrosis, cirrhosis, and hepatocellular carcinoma. Carbon tetrachloride (CCl4) treatment induced IL-20 that further up-regulated the expression of transforming growth factor (TGF)-β1 and p21WAF1 and resulted in cell cycle arrest in the Clone-9 rat hepatocyte cell line.

IL-20 activated quiescent rat hepatic stellate cells (HSCs) and HSP inhibitor up-regulated TGF-β1 expression. IL-20 also increased TGF-β1, tumor necrosis factor (TNF)-α, and type I collagen (Col-I) expression, and promoted the proliferation and migration of activated HSCs. Serum IL-20 was significantly elevated in mice with short-term and long-term CCl4-induced liver injury. In mice with short-term liver injury, anti-IL-20 monoclonal antibody (7E) and anti-IL-20 receptor (IL-20R1) monoclonal antibody (51D) attenuated hepatocyte damage caused by CCl4, TGF-β1, and chemokine production. In mice with long-term liver injury, 7E and 51D inhibited CCl4-induced cell damage, TGF-β1 production, liver fibrosis, HSC activation, and extracellular matrix accumulation, which

was caused by the reduced expression of tissue inhibitors of metalloproteinases check details as well as increased metalloproteinase expression and Col-I production. IL-20R1-deficient mice were protected from short-term and long-term liver injury. Conclusion: We identified a pivotal role of IL-20 in liver injury and showed that 7E and 51D may be therapeutic for liver fibrosis. (Hepatology 2014;60:1003–1014) “
“We tested the hypothesis that the pathogenesis of alcoholic liver injury is mediated by epigenetic changes in regulatory genes that result from the induction of aberrant methionine metabolism by ethanol feeding. Five-month-old cystathionine beta synthase heterozygous and wild-type C57BL/6J littermate mice were fed liquid control or ethanol diets by intragastric infusion for 4 weeks.

Of the 185 patients, 89 had serum samples stored in the serum bank. These samples were retrieved for virological analysis. Regression analysis indicated that the HBV-DNA levels derived from liver samples were significantly correlated with those from serum samples (regression coefficient 0.0439; 95% CI 0.0391-0.0487; P < 0.001) (Fig. 5). In a subgroup of patients, a large amount of HBV-DNA was found in the liver tissue, but a relatively lower HBV-DNA concentration was detected in the serum sample (Fig. 5, squares [n = 8]), suggesting a defect of viral secretion. In another subgroup of patients, a small amount of

HBV-DNA Epacadostat in vitro was found in the liver tissue, but a high HBV-DNA concentration click here was detected in the serum sample (Fig. 5, circles [n = 5]), indicating an extraordinarily high efficiency of viral secretion. Using cases without abnormal secretion efficiency, a regression equation was obtained to convert tissue to serum HBV-DNA levels for practical

application (Fig. 5). Sequence analysis for precore stop codon mutation, BCP mutation, and genotype revealed discrepancies between serum-derived and tissue-derived data in 11 patients, including eight and seven discrepancies for precore stop codon mutation and BCP mutation, respectively. Interestingly, of these 11 patients, six were in either of the two abnormal subgroups (Fig. 5, solid squares and circles). Discrepancies in the detection selleck chemicals llc of large fragment pre-S mutants occurred in 18 patients. The mutants were detected in the serum but not the tissue samples in 11 of them, whereas in the remaining seven patients, the mutants were detected in the tissue but not the serum samples. Five of these seven patients were in the subgroup with secretion defect (Fig. 5, asterisks).

Of the 89 patients with available serum samples, short fragment pre-S deletion mutants were detected in the liver tissues in nine of them (PSMUT 1-8 and 11). Of these mutants, seven were also detected in the serum samples (PSMUT 1-5, 7, 8). The remaining two were in the subgroup with secretion defect (Fig. 5, pound symbols). Despite HCC’s multifactorial etiology, HBV remains the major causative factor in Southeast Asia, where HBV is highly prevalent.26 By characterizing the HBV genome sequences derived from patients’ serum, several studies have illustrated that various virological factors are closely associated with hepatocarcinogenesis, including serum HBV-DNA concentration, genotype, and the presence of a basal core promoter mutation.27 Furthermore, by performing cell-based and animal-based experiments, several viral proteins have been shown to be implicated in the oncogenesis of liver cancer, including X proteins, large surface proteins, and pre-S deletion mutants.

36 Emu Oil lotion enhanced these parameters twofold, whereas pure Emu Oil did not exert significant effects.36 Improved wound healing with Emu Oil lotion was proposed to have occurred through a mechanism of enhanced keratinization.36 Nevertheless, in a study by Lagniel and Torres, Emu Oil improved recovery of damaged skin in children with second- and third-degree burn injuries caused by fire and hot water.37 The mechanism of action of Emu Oil and the nature of the active factor(s) are yet to be fully elucidated. It has been suggested that the n-3 and n-9 FAs present in

Emu Oil may confer anti-inflammatory learn more properties,31 and efforts to ameliorate several chronic inflammatory diseases, including IBD and rheumatoid arthritis, have been directed towards increasing dietary intake of n-3 and n-9 FAs.18,38 n-3 FAs reduce inflammation both directly (via downregulation of the inflammatory eicosanoid pathways that produce thromboxane B2, prostaglandin selleck kinase inhibitor E2 and leukotriene B4) and indirectly (by altering the expression of inflammatory genes through effects on transcription factor activation),39 whereas n-9 FAs inhibit macrophage migration.31 Yoganathan et al.31 demonstrated that the ability of Emu Oil to reduce levels of pro-inflammatory cytokines were more pronounced in an experimental model of inflammation, compared with other oils known to contain higher levels of FAs. This suggests that the

anti-inflammatory properties of Emu Oil could not be solely attributed to the FA profile alone.31 It is proposed that the effects of Emu Oil may

be attributed to the synergism of FAs and other constituents in Emu Oil and/or the FA ratio. Other minor constituents of Emu Oil in the non-triglyceride fraction, such as antioxidants including carotenoids and flavones, and skin-permeation enhancing factors, are reported to evoke antioxidant or radical scavenging activities,29 modulate anti-inflammatory, pro-apoptotic selleck chemicals and anti-proliferative pathways in intestinal epithelial cells40 and reduce pro-inflammatory cytokine production and colonic neutrophil infiltration in a mouse model of colitis.41 Furthermore, the high ratio of unsaturated to saturated fatty acids (UFA: SFA, approximately 1.8) may confer protection against oxidative damage.29 Emu Oil requires extensive testing, both topically and orally, with respect to its reported therapeutic benefits. Only in recent years have more rigorous studies of Emu Oil been conducted in pre-clinical models of gut disease. Lindsay et al.26 proposed a potential mechanism of action of Emu Oil following oral administration to rats with chemotherapy (5-Fluorouracil; 5-FU)-induced mucositis. In this study, Emu Oil decreased acute small intestinal inflammation assessed by myeloperoxidase activity (found in the intracellular granules of neutrophils) 96 h after 5-FU-administration.

With that said, this is comparable to other well-established screening initiatives that exist in the United States, such as cervical cancer or cholesterol screening. The limitations of this study are mostly intrinsic to its design. Because it is a model simulation, assumptions have to be made. These assumptions may be close NVP-LDE225 solubility dmso to, or veer far from, reality. For example, the probability of sustained virological response

(SVR) with DAAs plus standard of care was estimated based on results of one clinical trial (ADVANCE).7 This trial used telaprevir, one of the two approved PIs, and led, among previously naïve patients, to the highest SVR rate of 75%. This percentage was multiplied by the ratio of the average SVR rate of Peg-IFN/RBV therapy (genotypes 1/4) in primary care setting divided by the Selleckchem AZD3965 SVR of Peg-IFN/RBV therapy observed in clinical trials (0.33:0.46). As we all know, the real-world response rates will undoubtedly be less than 75%, in part as the result of the higher proportion of patients with cirrhosis that will be treated, with cirrhosis being a clear negative predictive factor of response

with triple therapy. There is no final data yet, but early data from the European Association for the Study of Liver Disease suggest, in patients with cirrhosis at least, lower response rates and more side effects, potentially leading to a higher discontinuation rate.10, 11 The assumed probability of SVR of 54% in the present study may or may not represent the real-life setting. A study published by McGarry et al. in HEPATOLOGY this year showed similar results.12 Also, based on a Markov model of the natural history of HCV, the investigators assessed the potential costs and benefits of a birth-cohort screening program in the United States. In this model, screening 100% of U.S. residents born 1946-1970 over 5 years would avoid 78,000 HCV-related deaths, which is analogous to the data in the Rein et

al. study. Similarly, the ICER of birth-cohort screening with DAAs plus standard treatment was $37,700 per QALYs saved, compared with risk-based screening, which is similar to the findings of Rein et al. As the anniversary of the approval of DAAs approaches, the CDC has proposed “an expansion of its current learn more risk-based guidelines to include a simple, one-time blood test for all baby boomers.”1 The cost-effectiveness analysis presented supports these recommendations. The investigators were wise to point out that these numbers are based on the published clinical trial data, which may overestimate the cure rates. On the other hand, at the pace at which HCV drug development is moving with the expected approval of two to four new DAAs in 2014 and many more after that, these lower real response rates may be a thing of the past.

Sclerostin mRNA in the liver was overex-pressed as compared with control samples (2.7±0.3 fold vs healthy liver). Sclerostin was detected by immunohistochemistry in 7 of the 11 liver samples but not in controls, and mainly located in the bile ducts. Sclerostin was directly Autophagy activator associated with the severity of cholangitis (p=0.02)

and indirectly with the degree of lobular inflammation (p=0.03), but not with the degree of fibrosis neither with the Ludwig’s histologic stage. Sclerostin mRNA expression was higher in samples positive by immunohistochemistry (2.9 ±0.4vs 2.5 ± 0.3, p: n.s.), and particularly in those with lobular granuloma (3.6±0.6 vs 2.4±0.2, p=0.02). Conclusion: The increased expression of sclerostin in the liver and the association with histologic cholangitis may explain the high serum levels of this protein in patients with primary biliary cirrhosis, thus suggesting that sclerostin influences the decreased bone formation in this cholestatic disease. Disclosures: Albert Pares – Consulting: Lumena Pharmaceuticals The following people have nothing to disclose: Silvia Ruiz-Gaspa, Laia Gifre, Nuria Guañabens, Rosa Miquel, Pilar Peris, Ana Monegal [Background and

Aim] Drug-induced cholestasis is a rare, but sometimes, fatal disease. Underlying mechanisms remain unknown. Organic anion transporting polypeptides OATP1B1 (gene: SLCO1B1) and OATP1B3 (gene: SLCO1B3) were identified as responsible Napabucasin datasheet click here for Rotor syndrome. We have recently demonstrated that six Japanese

patients with Rotor syndrome were homozygous for the insertion of LINE-1 (L1) retrotransposon in intron 5 of SLCO1B3 and for c.1738C>T (p.R580X) nonsense mutation in SLCO1B1. Intronic L1 insertion in SLCO1B3 causes skipping of exon 5 or exons 5-7 in SLCO1B3 mRNA, yields immature stop codon, and results in the generation of truncated OATP1B3 protein. OATP1B1 and 1B3 are involved in hepatic uptake of bilirubin glucuronides, bile acids, and steroidal and thyroid hormones, as well as various drugs such as statins and anticancer drugs. Therefore, their genetic abnormalities may cause acquired cholestasis. In this study we analyzed L1 insertion in SLCO1B3 and c.1738C>T mutation in SLCO1B1 in Japanese patients with drug-induced cholestasis. [Patients and Methods] A total of 44 Japanese patients with drug-induced cholestasis were enrolled after written informed consent was obtained. Inclusion criteria were (1) alkaline phos-phatase (AP) greater than 2 times the upper limit of normal (ULN), (2) R ≤ 2, when R was defined as alanine aminotransferase (ALT)/ULN divided by AP/ULN, and (3) absence of other hepatobiliary diseases. A single-tube, three-primer PCR assay was established to detect L1 insertion in SLCO1B3. The c.1738C>T mutation in SLCO1B1 was genotyped by direct sequencing. Allele frequency was compared with Japanese controls (n=554). [Results] Median (min-max) age of the cases was 65 (21-80) years and males were dominant (64%).

Stock

solutions of esterase (E3019; Sigma Inc., St. Louis, MO, USA), horseradish peroxidase (Sigma P8125), and catalase were prepared in DI water daily. The reaction mixture was inverted to mix and incubated for 3 min at room temperature before reading on a Fluoromax 2 fluorometer (HORIBA MK-8669 cell line Jobin Yvon Inc., Edison, NJ, USA). Excitation and emission were 488 and 525 nm, respectively. The experiment was conducted under low light conditions to avoid photooxidation of DCFH during sample incubation. Fluorescence readings in counts per min were converted to concentration of strong oxidants by comparison to a standard curve generated daily using H2O2. The concentration of strong oxidants was converted to the number of moles of oxidants released per gram of algal fresh weight (nm oxidants · g−1 FW) and strong oxidant release immediately upon wounding was determined by subtracting the amount of oxidants measured in

the seawater medium before wounding from those measured in the medium 1 min after wounding. The ability of 500 U catalase to decompose 1 M H2O2 in ice-cold seawater buy Seliciclib was verified each day by measuring DCFH fluorescence of a dilution of 30% H2O2 (Sigma 216763) made in ice-cold SFSW as above with and without the addition of 500 U catalase from the fresh catalase stock solution. In order to look at the relative timeline of the oxidative burst, we sampled oxidant release in the seawater medium over the course of 65 min after the wounding of three species (A. mirabilis, P. decipiens, and T. antarcticus). Samples were wounded at t = 5 min and oxidants were measured as above, with and without the addition of 500 U catalase, in the medium of each species (n = 3) at times 0, 20, 40, and 70 min. To determine whether RNS were a component of the wound-induced oxidative burst, paired samples of A. mirabilis, D. anceps, P. decipiens, and T. antarcticus were flash frozen in liquid nitrogen 30–60 s after punching with a sterile pipette tip. Protein was extracted from each

sample using a plant total protein extraction kit (Sigma PE0230) and quantified using a Bradford-based selleck protein microassay (Bio-Rad protein assay kit II #500-0002), both according to the manufacturer’s protocol. For comparison, protein was also extracted from samples of Saccharina latissima (Linnaeus) C.E. Lane, C. Mayes, Druehl et G.W. Saunders collected from the dock at Friday Harbor Laboratories (University of Washington, Friday Harbor, WA 98250, USA) in August 2012 and exposed to peroxynitrite (ONOO−, Cayman Chemical 81565, Ann Arbor, MI, USA), an RNS and a strong nitrating agent. Protein samples were stored at −80°C until use. Nitrated proteins were detected using 1-D 10% SDS polyacrylamide gel electrophoresis (PAGE) with subsequent transfer to PVDF membrane and immunoblotting. 20–40 μg of protein were loaded for 1-D PAGE depending on the species, with equal amounts loaded for each replicate within a species (n = 6 for P.