Samples were collected one day prior to laboratory procedures and stored overnight in a domestic refrigerator (5°C) prior to processing. For each sample, microbiological and molecular analyses were conducted on both intact (unsterilized) material and on surface sterilized material. Unsterilized samples (an assortment of leaves corresponding to 10–20 g of leaf material) were washed under regular tap water (as might be done by a typical consumer) and then added to bottles containing 100 mL of sterile magnesium phosphate buffer [40]. Surface sterilized samples (10–20 g of leaf material) were washed in the

same manner as unsterilized samples and then placed into sterile sample bottles. These bottles then received #SC79 cost randurls[1|1|,|CHEM1|]# 100 ml of a 1.3% sodium hypochlorite solution and were shaken (200 rpm) for 5 min. The sodium hypochlorite solution was decanted and replaced with 70% ethanol, and bottles were shaken for a further 2 min. The ethanol was decanted, replaced with 100 ml sterilized distilled water, learn more and bottles were shaken for 10 seconds. The water was removed and this sterile water rinse repeated three more times to ensure that there was minimal sodium hypochlorite or ethanol remaining in the bottle. Following the final wash, 100 mL of sterile magnesium phosphate buffer was added to the bottle. Efficiency of this sterilization technique

was tested by wiping of sterilized leaves of each type across the surface of a trypticase soy agar (TSA) plate, which consistently yielded no bacterial colonies. Culture dependent microbiological analyses Surfaced sterilized and unsterilized samples were homogenized using a Power Gen 500 homogenizer Forskolin ic50 (Fisher Scientific) and the resulting leaf slurries serially diluted ten-fold. Subsamples (0.1 mL) of each dilution were plated in triplicate onto both TSA and R2A agar; each medium also contained 0.1 g L-1 cycloheximide to inhibit fungal growth. Plates were incubated at room temperature (22°C) for 2–5 d, after which time colonies were counted

and final counts expressed as CFU g-1 leaf vegetable. Colonies were qualitatively typed based on color and overall morphology, and a sample of each numerically dominant morphological colony type was transferred onto a new plate of the appropriate medium and incubated (22°C; 2–4 d). These isolates were transferred three times to ensure purity. Following growth of the third transfer, DNA was extracted from a single colony of each isolate using UltraClean Microbial DNA Isolation kits (Mo Bio Laboratories, Carlsbad, CA). A portion of the 16S rRNA gene was amplified using the Bac799f and Univ1492r primers with amplification conditions described below and amplicons subsequently sequenced. Potentially erroneous bases (low quality scores) were removed and sequences were then processed through the Greengenes database [41] in order to identify and classify them.

This process is primarily a function of vasodilation of the arterioles (distal, proximal, and feed) and the pre-capillary sphincters, which is to a great degree induced by factors such as adenosine, carbon dioxide, and potassium, which are released in proportion to intensity of effort by adjacent muscle fibers during exercise [4]. The close coupling of muscular blood flow and exercise intensity supports the theory that further elevations in localized blood flow during exercise may, in some cases, result in increased peak work capacity and/or increased resistance to local muscle fatigue, thereby enhancing exercise performance. The process of vasodilation

NVP-BSK805 cell line as a primary component of exercise hyperemia involves mechanisms other than the aforementioned muscle metabolite induced vasodilatory mechanisms (adenosine, CO2, K+). For example, the initial increases of blood flow (first 1 – 2s) during exercise are now believed to be related to increased concentrations of acetylcholine

as released by the motor end-plate during muscle activation [5]. Tschakovsky and Joyner [6] outlined several mechanisms believed to contribute to the secondary phase of vasodilation (3+ sec) including flow mediated mechanisms, the mechanical muscle pump, mechanically induced responses, muscle activation Erismodegib mechanisms, and red blood cell HbO2 desaturation mechanisms. Each of these mechanisms can be associated with during different variations and intensities of exercise stresses. However, each of these distinct mechanisms shares the common function of initiating the synthesis of nitric oxide (NO). Nitric oxide (NO) is a very short-lived, reactive gaseous nitrogen molecule that is involved in a variety of physiological functions. Approximately twenty years ago, it was revealed that NO was the endothelial factor responsible for regulating muscle tone of vascular

structures, originally referred to as endothelial dependent relaxation factor (EDRF) several years prior. However, a viable means to manipulate this molecule has not been identified. Therefore, it is uncertain at this time what influence increased production of NO would have on cardioselleck chemicals llc vascular functioning and/or resistance to local muscle fatigue. Nitric oxide is synthesized in endothelial cells from arginine via enzymatic action of endothelium nitric oxide synthase. This molecule diffuses easily into the vascular smooth muscle where it binds to the enzyme guanylyl cyclase, which in turn catalyzes the phosphorylation of gunaosine-5-triphosphate (GTP) into cyclic gyanosine monophosphate (cGMP). Cyclic GMP serves as an important second messenger for many physiological functions, including relaxation of smooth vascular muscle. The amino acid, arginine, acts as a precursor to NO synthesis. Due to this role, a significant nutritional supplement market has developed for arginine-based products which supposedly enhance the production of NO.

The results were expressed as the mean value of at least ten pendant drops at 23°C and 55% relative humidity. Biosurfactant serial dilutions BIRB 796 mouse in water were performed and analyzed using the pendant drop technique described above to determine the critical micellar concentration [34]. The measurements were taken until the Volasertib nmr surface tension was close to the one of water. Analysis of conditioned surfaces The surfaces samples were 2 cm2 coupons of stainless steel AISI 304, stainless steel AISI 430, carbon steel, galvanized steel and polystyrene. All of

them were cleaned by immersing them in 99% ethanol (v/v), placing them in an ultrasonic bath for 10 min, rinsing them with distilled water, immersing them in a 2% aqueous solution of commercial detergent and ultrasonic cleaning them for 10 more minutes. The coupons were washed with CBL-0137 distilled water and

then sterilized at 121°C for 15 min. The cleaned coupons were then conditioned with aqueous solutions 5% (w/v) of the dried powder obtained after neutralization of AMS H2O-1 lipopeptide extract, surfactin or water (control) by immersing them in the solutions for 24 h at room temperature. The samples were then washed with water and left to dry at room temperature until further analysis. The water, formamide and ethylene glycol drop angles were measured to determine the surface free energy and hydrophilic and hydrophobic characteristics of the metal and non-metal surfaces after they were conditioned

with the AMS H2O-1 lipopeptide extract, surfactin, or water (control). The assays were performed using a Krüss DSA 100S goniometer (model: OF 3210) to measure the contact angles between the liquids and the different surfaces (stainless steel AISI 304, stainless steel AISI 430, carbon steel, galvanized steel and polystyrene). The results are expressed as the mean value of at least ten drops (10 μl) at 23°C and 55% relative humidity. The surface free energy was calculated from the surface tension components from each known liquid obtained from the Cyclooxygenase (COX) contact angle using the equation 1 [35]: (1) where: θ is the contact angle between the liquid and the surface; γTOT is the total surface free energy; γLW is the Lifshitz-van der Waals component; γAB is the Lewis acid–base property; γ+ and γ- are the electron acceptor and donor components, respectively; . The surface hydrophobicity was determined through contact angle measurements and by the approach of Van Oss [35] and Van Oss et al. [36], which states that the degree of hydrophobicity of a material (i) is expressed as the free energy of the interaction between two entities of that material when immersed in water (w), ΔGiwi. If the interaction between the two entities is stronger than the interaction of each entity with water, the material is considered hydrophobic (ΔGiwi<0). Hydrophilic materials have a ΔGiwi>0.

In contrast, treatment with the cytostatic drug cyclophosphamide prevents the recruitment of immune effector cells to the side of infection. Therefore, despite a retarded germination of conidia, fungal hyphae stay alive, which is well visualized by the massive increase in fungal DNA determined at the late stage of infection (Figure 2). In agreement, the bioluminescence steadily

increased under this regimen and explanted lungs show a 50 – 100 times higher light emission than observed under corticosteroid treatment. This result shows that bioluminescence measurements and DNA quantification correlate best under the cyclophophamide regimen. Although the bioluminescence readout does not correlate linearily with the fungal burden as measured by qRT-PCR, the general tendency of increasing and decreasing fungal burden as well as the impact of the inflammatory

response seems well reflected AZD1480 nmr by bioluminescence imaging. Impact of immunosuppression regimens on the inflammatory response In order to correlate survival curves, weight loss, fungal burden from DNA quantification and bioluminescence with histopathological findings, additional experiments were performed, in which mice were sacrificed one day (early) and three days (late) post infection. For the clodrolip condition, S63845 concentration mice were sacrificed eight days after infection to assess any later effect of treatment on mice survival. Lungs were removed, and thin sections were studied for the www.selleckchem.com/products/ly2606368.html evaluation of the recruitment of immune effector cell lineages and fungal tissue invasion. Clodrolip treatment Lung instillation with clodrolip was expected to reduce the number of AM, which are generally denoted as the first cellular line of host innate immune defense through phagocytosis and killing of inhaled conidia. To confirm the reduction in the number

of AM, the BAL Tacrolimus (FK506) fluid of non-infected mice were sampled two days after intranasal administration of clodrolip or liposomes, respectively. Flow cytometry was used to quantify the number of AM within the BAL fluid. The clodrolip treatment resulted in a numeric depletion of 60% of AM (8.30 × 104 ± 1 × 104 versus 2.03 × 105 ± 1.8 × 104) when compared to control liposome treated animals (p < 0.05). Furthermore, the viability of the residual AM subset was only 50% as evaluated by trypan blue staining. Taken together, clodrolip treatment depleted or resulted in the death of 80% of AM compared to control mice. When the cell populations in BAL were evaluated one day post-infection, we noted a 3.2-fold decrease (22 ± 11 versus 71 ± 28%) in the concentration of AM and a 2.6-fold increase (77.5 ± 10 versus 29 ± 28%) in the neutrophil concentration in clodrolip-treated mice compared to control liposome-treated mice (Figure 3A).

Switch to second-line antibiotic therapy was defined as the addition of one or more parenteral antibiotics to the initial antibiotic regimen

or as a complete or partial switch of the initial antibiotic regimen to another parenteral antibiotic regimen. Unscheduled additional abdominal surgeries were taken into account if they occurred 2 or more days after the primary surgical procedure and were related to poor primary source control. Secondary procedures were not considered in the analysis when there was a mention of other reasons (i.e. technical issues or hemorrhage) that might have led to re-operation. First-line empiric antibiotic therapy was defined as appropriate if all isolated bacteria were sensitive to at least one of the antibiotics administered CBL-0137 in patients with Akt inhibitor documented positive intra-abdominal swabs or blood cultures. Alternatively, in patients with negative or no cultures, empiric therapy was deemed as appropriate when the selected regimen covered enteric gram-negative aerobic and anaerobic bacteria and drug dosing was adequate, Tozasertib molecular weight according to current guidelines [1]. Antibiotic regimens not fulfilling the above criteria were defined as inappropriate. Leucocytosis was defined by a white blood cell (WBC) count >12,000/mm3. Leukopenia was defined as a WBC count <4000/mm3. Cost analysis A estimate of the cost of antibiotics was performed by multiplying the number of antibiotic

days by the unit price of that antibiotic and by the number of per day doses. The overall cost of antibiotic treatment for each patient was the sum of costs calculated for all parenteral antibiotics received by the patient during the hospitalization period. The unit price of antibiotics was based on official ex-factory prices per unit in Italy [12]. Laboratory tests, instrumental tests, and specialists’ consultancies utilization were directly recorded and their costs were assessed by referring to fees for providers of specialist services recognized by

the Italian National Health Service (I-NHS). Costs related to primary surgical procedures were not included in analysis, as we assume they were independent Demeclocycline of the adopted antibiotic therapy. Other direct costs, including personnel, ordinary maintenance and hotel costs, were indirectly estimated by using Diagnosis-Related Group’s tariffs per admission provided to hospitals by the I-NHS. Specifically, this estimate was based on the acknowledged over-threshold per hospital day tariff, which is the per day cost to hospitals for length of stay prolonged over an a priori defined threshold (i.e. a tariff applicable to length of stay statistically considered as outliers), assuming that by subtracting the average costs of specialist services provided from this tariff, an acceptable proxy of the general cost sustained for patient management could be obtained. Costs were expressed in Euro values at the time they were incurred (year 2009 values).

Lancet 2003, 361:1715–1722.PubMedCrossRef 2. Cheng AC, Currie BJ: Melioidosis: epidemiology, pathophysiology, and management. Clin Microbiol Rev 2005, 18:383–416.PubMedCrossRef 3. Currie BJ, Jacups SP: Intensity of rainfall and severity of melioidosis, Australia. Emerg Infect Dis 2003, 9:1538–1542.PubMed 4. Suputtamongkol Y, Hall AJ, Dance DA, Chaowagul Selleck Small molecule library W, Rajchanuvong A, Smith MD, White NJ: The epidemiology of melioidosis in Ubon Ratchatani, northeast Thailand. Int J Epidemiol 1994, 23:1082–1090.PubMedCrossRef 5. Leelarasamee A, Trakulsomboon S, Kusum M, Dejsirilert S: Isolation rates of

Burkholderia pseudomallei among the four regions in Thailand. Southeast Asian J Trop Med Public Health 1997, 28:107–113.PubMed 6. Vuddhakul V, Tharavichitkul P, Na-Ngam N, Jitsurong S, Kunthawa B, Noimay P, Noimay P, Binla A, EVP4593 ic50 Thamlikitkul V: Epidemiology of Burkholderia pseudomallei in Thailand. Am J Trop Med Hyg 1999, 60:458–461.PubMed 7. Wongpokhom N, Kheoruenromne I, Suddhiprakarn A, Gilkes RJ: Micromorphological properties of salt affected soils in Northeast Thailand. Geoderma 2008, 144:158–170.CrossRef 8. O’Quinn AL, Wiegand EM, Jeddeloh JA: Burkholderia

pseudomallei kills the nematode Caenorhabditis elegans using an endotoxin-mediated paralysis. Cell Microbiol 2001, 3:381–393.PubMedCrossRef 9. Vandamme P, Holmes B, Vancanneyt M, Coenye T, Hoste B, Coopman R, Revets H, Lauwers S, Gillis M, Kersters K, et al.: NADPH-cytochrome-c2 reductase Occurrence of multiple genomovars of Burkholderia cepacia in cystic fibrosis patients and proposal of Burkholderia multivorans sp. nov. Int J Syst Bacteriol 1997, 47:1188–1200.PubMedCrossRef 10. Mahenthiralingam E, Baldwin A, Vandamme P: Burkholderia cepacia complex infection in patients with cystic fibrosis. J Med Microbiol 2002, 51:533–538.PubMed 11. Widdicombe JH: Altered NaCl concentration of airway surface liquid in cystic fibrosis. Pflugers Arch 2001,443(Suppl

1):S8–10.PubMed 12. Joris L, Dab I, Quinton PM: Elemental composition of human airway surface fluid in healthy and GW786034 diseased airways. Am Rev Respir Dis 1993, 148:1633–1637.PubMed 13. O’Carroll MR, Kidd TJ, Coulter C, Smith HV, Rose BR, Harbour C, Bell SC: Burkholderia pseudomallei : another emerging pathogen in cystic fibrosis. Thorax 2003, 58:1087–1091.PubMedCrossRef 14. Choy JL, Mayo M, Janmaat A, Currie BJ: Animal melioidosis in Australia. Acta Trop 2000, 74:153–158.PubMedCrossRef 15. Dance DA: Ecology of Burkholderia pseudomallei and the interactions between environmental Burkholderia spp. and human-animal hosts. Acta Trop 2000, 74:159–168.PubMedCrossRef 16. Yamamoto T: [Stress response of pathogenic bacteria--are stress proteins virulence factors?]. Nippon Saikingaku Zasshi 1996, 51:1025–1036.PubMed 17. Pumirat P, Saetun P, Sinchaikul S, Chen ST, Korbsrisate S, Thongboonkerd V: Altered secretome of Burkholderia pseudomallei induced by salt stress. Biochim Biophys Acta 2009, 1794:898–904.PubMed 18.

Bacterial enumeration showed no differences between the two growth conditions, indicating that pGEN-lux is stable in vivo up to 96 hpi in all organs tested (Figure 1). Additionally, organs from all animals imaged in this study ACY-738 research buy were also plated on BHI and BHI with carbenicillin (after last imaging time point). We observed the same levels of plasmid stability that we report in Figure 1 (data not shown). Figure 1 Bacterial loads in C57Bl/6J mice infected subcutaneously with pGEN- luxCDABE

-carrying Y. pestis. Animals were infected with ~200 CFU at a cervical site. Organs were harvested and plated for bacterial counts at the indicated hours post inoculation on BHI alone (gray symbols) and BHI + carbenicillin (white symbols). Bacterial numbers are reported in CFU/g of tissue. Each mark represents MK-8931 order a value from a single organ and the horizontal lines represent the 4SC-202 datasheet median of the group. Superficial cervical lymph nodes are represented as circles, spleens as squares, and lungs as triangles. A dotted line represents the limit of detection. Data shown from a single

experiment. Another important control experiment was to determine if pGEN-lux impacted the virulence of Y. pestis. Mice were inoculated with either Y. pestis alone or Y. pestis carrying pGEN-lux. Both groups of mice displayed signs of plague infection and mortality at similar times. However, the bacterial burden in tissues from mice infected with Y. pestis carrying pGEN-lux was lower in comparison to tissues from mice infected with Y. pestis without the plasmid (Figure 2). While bacterial counts suggest that pGEN-lux might cause a slight delay in the progression of infection, overt signs of plague were observed in all mice infected with either strain at comparable times. Additionally, all mice infected during our BLI experiments died at times expected from infections with a wild type strain. Since all strains used for BLI will carry

the same plasmid, relative virulence attributes will be comparable despite the slight attenuation caused by BCKDHA pGEN-lux. Figure 2 Bacterial loads in C57Bl/6J mice infected subcutaneously with either wild type or pGEN- luxCDABE -carrying Y. pestis. Animals were infected with ~200 CFU at a cervical site. Organs were harvested and plated for bacterial counts at the indicated hours post inoculation. Bacterial numbers are reported in CFU/g of tissue. Gray and white symbols represent organs from animals infected with Y. pestis and Y. pestis carrying pGEN-luxCDABE, respectively. Each mark represents a value from a single organ and the horizontal lines represent the median of the group. Superficial cervical lymph nodes are represented as circles, spleens as squares, and lungs as triangles. A dotted line represents the limit of detection and an x letter represents missing values of a specific tissue due to the death of an animal. Data shown from a single experiment. BLI of Y.

Of course, this suggested approach is similar to previous attempts to separate https://www.selleckchem.com/products/pci-34051.html phytoplankton groups based on fluorescence excitation spectra (Millie et al. 2002; Beutler et al. 2002; Beutler et al. 2004; Parésys et al. 2005; Gaevsky et al. 2005; Seppälä and Olli 2008). The small number of algal and cyanobacterial species used in our this website experiments, despite being grown in conditions to allow for a wide range in F v/F m, limits the applicability of our

results. Fluorescence emission profiles of the major algae groups are relatively similar because the main source of fluorescence is always Chla located in PSII. The excitation spectrum, on the other hand, is dependent on the accessory photosynthetic pigments present. The choice of a single chlorophyte and diatom, representing red absorption by Chlorophylls b and c, is therefore still a realistic representation of many natural communities where algae and cyanobacteria co-exist. LY3023414 in vitro It does, however, not cover natural communities extensively. We may

consider the case of phycobilin-producing rhodophytes and cryptophytes, as well as cryptophyte-ingesting ciliates (Gustafson et al. 2000) in further studies. The fluorescence excitation–emission matrices of rhodophytes are similar to those of the cyanobacteria used here, although planktonic rhodophytes are generally few in environments where cyanobacteria are abundant. We hypothesize that the solutions for instrument design proposed here apply to these algae in the same manner as for the cyanobacteria described here. In contrast, the presence of phycoerythrin in cryptophytes and some dinoflagellates leads to a broader excitation domain in the algal groups. The presence of these ‘special’ algal groups in a natural sample will hamper efforts to decompose multi-channel fluorescence measurements into

the contributions by individual groups (but see Seppälä and Olli buy Gefitinib 2008), even though it should not markedly change our definition of optimal excitation–emission bands to yield results that are most representative of the whole phytoplankton community. The PBS pigments produced by strains in this study absorb yellow-to-red light as is common to freshwater and coastal species. The presence of oceanic species with forms of phycoerythrin absorbing down to 495 nm (Lantoine and Neveux 1997; Subramaniam et al. 1999; Neveux et al. 2006) would reduce the specificity of the blue-excited fluorescence signals to the algal part of the community, but we remain confident that the inclusion of an orange-to-red excitation band markedly increases sensitivity to the cyanobacteria present.

Despite the automatic annotations, all the gene findings in this study were based on manual gene comparison rather than automatic annotation, since in several cases the automated annotation was incorrect. In order to determine whether a gene has homologs existing in other genomes, we used the genomic BLAST tool of the NCBI [68] with the tblastn (search translated nucleotide database using

a protein query) algorithm for searching. The Genome-To-Genome Distance Calculator [69] was used for genome-based species delineation as described [70]. This system calculates DNA-DNA similarity values by comparing the genomes to obtain high-scoring segment pairs (HSPs) and inferring distances from a set of three formulas (1, HSP length/total length; 2, identities/HSP length; 3, identities/total length). Spectroscopic DNA-DNA reassociation experiments were

performed according to the protocol outlined by the DSMZ Identification Service [62]. selleck chemicals llc Phylogenetic trees based on 16S rRNA, pufLM and rpoB gene sequences were reconstructed using distance matrix (neighbor-joining) and buy CH5183284 parsimony programs included in the ARB package [71]. Maximum likelihood trees were reconstructed with the program RAxML (version 7.2.8) using raxmlGUI [72] and the GTRGAMMA option with 1000 rounds of bootstrap replicates [73]. The dataset of aligned and almost complete 16S rRNA gene sequences was based on the ARB SILVA database release 108 (September 2011) [74], whereas DNA sequences of pufL, pufM and rpoB genes were 5-Fluoracil cell line obtained from GenBank and aligned using the ClustalW algorithm implemented in the ARB package. The generated alignments of pufLM and rpoB this website nucleotide sequences in PHYLIP format are available as Additional file 2 and Additional file 3, respectively. Identity values of aligned nucleotide sequences were determined by using the similarity option of the neighbor-joining program included in the ARB package. Acknowledgements We thank Ivalyo Kostadinov and Alexandra Meziti for taking of samples. We are grateful to the Genome Analytics group (HZI Braunschweig) for providing sequence data

of DSM 19751T and to Anne Fiebig (DSMZ Braunschweig) for help with the genome assembly. The assistance of Andrey Yurkov (DSMZ Braunschweig) in performing maximum likelihood analyses is gratefully acknowledged. The excellent technical assistance of Jörg Wulf (MPI Bremen), Nicole Mrotzek, Gabriele Pötter and Bettina Sträubler (all DSMZ Braunschweig) is acknowledged. We are grateful to Dr. J. P. Euzéby (http://​www.​bacterio.​net/​) for correcting the etymology of the proposed Latin name of strain Ivo14T and to Dr. B. T. Tindall (DSMZ Braunschweig) for helpful discussions. TR was supported by the DFG Transregio-SFB 51 Roseobacter. BMF and SY were supported by the Max Planck Society. Genome sequencing of strains Ivo14T and Rap1red was funded by the Marine Microbiology Initiative of the Gordon and Betty Moore Foundation.

09). This indicated that the null AZD4547 solubility dmso association of C282Y and HCC when compared in HCC cases and viral LC cases should be taken with caution and that it warranted further study in a larger scale. FPRP is a valuable criterion to assess whether or not a positive discovery came about by chance. We used FPRP to assess the positive association attained by this meta-analysis. The association between C282Y (Y vs. C) and HCC attained by subgroup analysis of four studies using alcoholic LC patients

as controls was proved to be reliable (FPRP = 0.03). Population-attributable risk (PAR) is a valuable parameter to assess the influence of risk factors on disease occurrence. The PAR of the variant allele Y of C282Y among alcoholic LC patients was 5.12% (95%CI: 2.57%-7.67%). This result suggested that the role 4SC-202 of C282Y polymorphism on HCC occurrence was modest. Conclusions This meta-analysis proved that C282Y mutation was associated with HCC in European alcoholic LC patients. The role of C282Y polymorphism on HCC occurrence was modest. The association of this polymorphism and HCC is warranted further studies in large scale including diverse ethnicities.

The molecular mechanism 3 Methyladenine of the different effect of C282Y on alcoholic LC and viral LC, with respect to HCC occurrence, also merits further studies. This meta-analysis did not find association of H63D mutation with HCC. Acknowledgements The present study was supported by the China Ministry of Health (2009ZX10004-301), National Natural Science Foundation (No. 30772505, No. 30872503 & No. 40830744), National Basic Research Program of China (2007CB936004) and China National Key Projects for Infectious Diseases (2008ZX10002-017). References 1. Mazzaferro V, Llovet JM, Miceli R, Bhoori S, Schiavo M, Mariani L, Camerini

T, Roayaie S, Schwartz ME, Grazi GL, Adam R, Neuhaus P, Salizzoni M, Bruix J, Forner A, De Carlis L, Cillo U, Burroughs AK, Troisi R, Rossi M, Gerunda GE, Lerut J, Belghiti J, Boin I, Gugenheim J, Rochling F, Van Hoek B, Majno Amino acid P: Predicting survival after liver transplantation in patients with hepatocellular carcinoma beyond the Milan criteria: a retrospective, exploratory analysis. Lancet Oncol 2009,10(1):35–43.PubMedCrossRef 2. Edwards CQ, Dadone MM, Skolnick MH, Kushner JP: Hereditary haemochromatosis. Clin Haematol 1982,11(2):411–435.PubMed 3. Tavill AS: Diagnosis and management of hemochromatosis. Hepatology 2001,33(5):1321–1328.PubMedCrossRef 4. Niederau C, Fischer R, Sonnenberg A, Stremmel W, Trampisch HJ, Strohmeyer G: Survival and causes of death in cirrhotic and in noncirrhotic patients with primary hemochromatosis. N Engl J Med 1985,313(20):1256–1262.PubMedCrossRef 5. Fargion S, Mandelli C, Piperno A, Cesana B, Fracanzani AL, Fraquelli M, Bianchi PA, Fiorelli G, Conte D: Survival and prognostic factors in 212 Italian patients with genetic hemochromatosis. Hepatology 1992,15(4):655–659.PubMedCrossRef 6.