First, epidemiological studies have found that SS2 outbreaks are usually infrequent and only affect a small number of pigs, which can lead to underdiagnosis or misdiagnosis. Second, pigs infected with SS2 do not always show obvious clinical symptoms, and may become carriers without showing clinical signs. Finally, based on its polysaccharide capsular antigens, at least 35 serotypes of S. suis exist. Isolates belonging to other serotypes (such as 1, 1/2, 3, 4, 5, 7, 8 and 9) have also been associated with disease in pigs [28, 29]. Common

antigens had been found to be shared between SS2 and these other serotypes (unpublished data from our lab). To reduce these possible interferences, we used www.selleckchem.com/products/Tipifarnib(R115777).html pigs with clear backgrounds as animal models, and convalescent sera were prepared following artificial infection. Until recently, the exact mechanism of SS2 transmission (from LXH254 purchase pig to human or between pigs) was still poorly understood, but was thought to involve aerosol transmission or other pathways [28–30]. However, some hypotheses about the critical stages of the infection, such as bacterial invasion from the mucosal surfaces to the bloodstream, survival of the bacteria in blood, and

invasion from blood into the central nervous system have been presented [28]. Regardless of the mechanism of SS2 invasion, circulation in the blood plays an important role during SS2 disease development. In addition, S. suis is an agent of zoonosis, afflicting people in close contact Nintedanib with infected pigs or pork-derived products. The organisms probably gain entry via small wounds or through inhalation [4, 10, 29]. Furthermore, transmission between pigs in herds through cutaneous wounds has been suggested [29]. In light of

these considerations, intravenous and intramuscular inoculations were employed to assay the expression of SS2 in vivo, and to try to mimic natural infection (such as the middle or late stage of the infection). In this study, we used real-time PCR to analyze the induction of the expression of IVI genes under different environmental conditions. Real-time PCR results demonstrated that the expression of six of the 10 selected genes was upregulated under in vivo conditions. The upregulation time points for these six genes were 12, 24, and 36 h for ss-1616 and trag, 24 h for hprk and sdh, and 36 h for nlpa and ss-1298. This upregulated expression suggests that these genes may play a significant role during the course of SS2 infection (middle, late, or whole stage of infection). The expression profiles of the other four genes (ysirk, srt, cwh, and ss-1955) showed that they were not obviously upregulated under the in vivo condition (Figure 3). There are two possible explanations for this result. First, since we measured the in vivo gene expression at 12, 24, and 36 h pi, it is possible that we missed the time when the levels of expression of these genes were high relative to the expression of the same gene in vitro.

A personal responsibility to disclose genetic information A personal responsibility to

disclose genetic information is more permissive in describing what we expect to happen in family check details relationships, as opposed to a legal obligation, which is more about what we require. In this instance, it permits a patient to decide what, to whom, when, and how to disclose information that could have an impact on the health of a family member, as well as on the family member’s relationship with the patient. The familial context of each patient is different (Wiseman et al. 2010), and a personal responsibility recognizes this. This responsibility has adherents in national and international guidelines and policies that promote patient disclosure of genetic risk to their families. Although these are often not detailed, they are a starting point for discussion. In Germany, a personal responsibility to communicate genetic risk is explicit. “A moral obligation of family members to share their knowledge of their genetic makeup can be seen, as well as a moral obligation of partners to inform each other of their medical genetic problems, insofar as the latter concern children they may have in common” (German Society of Human Genetics 1998). France also takes a more explicit view of the obligation MK-1775 cell line of patients. The National Consultative

Ethics Committee for Health and Life Sciences makes clear that the patient has the moral responsibility (though not the legal) to disclose pertinent information to those who could benefit (France National Consultative Ethics Committee for Health and Life Sciences (CCNE) 2003). In the UK, the General Medical Council recognizes that most patients will share genetic information with relatives if properly advised of the health implications of the

information (General Medical Council 2009). The Nuffield Council on Bioethics is clear that patients “acting responsibly would normally wish to communicate important genetic information to other family members who may Bacterial neuraminidase have an interest in that information, and… that the primary responsibility for communicating genetic information to a family member or other third party lies with the [patient] and not with the doctor who may, however, do this at the request of the person concerned” (Nuffield Council on Bioethics 1993). This statement places responsibility for disclosure solely with the patient, though it does not provide further direction as to how and when patients should do so. Finally, the Joint Committee on Medical Genetics recently released guidance on consent in genetic practice, emphasizing the importance that genetic information might hold for family members and recognizing the patient as a potential source of the disclosure (Royal College of Physicians et al. 2011). Other guidance implies a responsibility for patients to inform family of risk.

Nat Rev Drug Discov 2012, 11:37–51. 18. Seo MD, Won HS, Kim JH, Mishig-Ochir T, Lee BJ: Antimicrobial peptides for therapeutic applications: a review. Molecules 2012, 17:12276–12286.PubMedCrossRef 19. Campbell Belinostat supplier EL, Serhan CN, Colgan SP: Antimicrobial aspects of inflammatory resolution in the mucosa: a role

for proresolving mediators. J Immunol 2011, 187:3475–3481.PubMedCrossRef 20. Lehrer RI, Lu W: alpha-Defensins in human innate immunity. Immunol Rev 2012, 245:84–112.PubMedCrossRef 21. Mehra T, Koberle M, Braunsdorf C, Mailander-Sanchez D, Borelli C, et al.: Alternative approaches to antifungal therapies. Exp Dermatol 2012, 21:778–782.PubMed 22. Zhu S: Discovery of six families of fungal defensin-like peptides provides insights into origin and evolution of the CSalphabeta defensins. Mol Immunol 2008, 45:828–838.PubMedCrossRef 23. Batoni G, Maisetta G, Brancatisano FL, Esin S, Campa M: Use of antimicrobial peptides against microbial biofilms: Epigenetics Compound Library concentration advantages and limits. Curr Med Chem 2011, 18:256–279.PubMedCrossRef 24. Dziarski R, Gupta D: Review: Mammalian peptidoglycan recognition proteins (PGRPs) in innate immunity. Innate Immun 2010, 16:168–174.PubMedCrossRef 25. Taraszkiewicz A, Fila G, Grinholc M, Nakonieczna J: Innovative strategies

to overcome biofilm resistance. Biomed Res Int 2013, 2013:150653. doi: 10.1155/2013/150653PubMed 26. Cota-Arriola O, Cortez-Rocha MO, Burgos-Hernandez A, Ezquerra-Brauer JM, Plascencia-Jatomea M: Controlled release matrices and micro/nanoparticles of chitosan with antimicrobial potential: development of new strategies for microbial control in agriculture. J Sci Food Agric 2013, 93:1525–1536.PubMedCrossRef 27. Dhople V, Krukemeyer A, Ramamoorthy A: The human beta-defensin-3, an antibacterial peptide with multiple biological functions. Biochim Biophys Acta 2006, Resminostat 1758:1499–1512.PubMedCrossRef 28.

Joly S, Maze C, McCray PB Jr, Guthmiller JM: Human beta-defensins 2 and 3 demonstrate strain-selective activity against oral microorganisms. J Clin Microbiol 2004, 42:1024–1029.PubMedCrossRef 29. Mooney C, Haslam NJ, Pollastri G, Shields DC: Towards the improved discovery and design of functional peptides: common features of diverse classes permit generalized prediction of bioactivity. PLoS One 2012, 7:e45012.PubMedCrossRef 30. Na DH, Faraj J, Capan Y, Leung KP, DeLuca PP: Stability of antimicrobial decapeptide (KSL) and its analogues for delivery in the oral cavity. Pharm Res 2007, 24:1544–1550.PubMedCrossRef 31. Hong SY, Park TG, Lee KH: The effect of charge increase on the specificity and activity of a short antimicrobial peptide. Peptides 2001, 22:1669–1674.PubMedCrossRef 32. Oh JE, Hong SY, Lee KH: Structure-activity relationship study: short antimicrobial peptides. J Pept Res 1999, 53:41–46.PubMedCrossRef 33. Concannon SP, Crowe TD, Abercrombie JJ, Molina CM, Hou P, et al.: Susceptibility of oral bacteria to an antimicrobial decapeptide. J Med Microbiol 2003, 52:1083–1093.PubMedCrossRef 34.

The results might be affected by an underlying selection bias due to the nature of retrospective data. Also, our study was limited by the small number of patients, the heterogeneity of the disease, the transplant procedure and the stem cell source. However, the major strengths of our study were that the follow-up period was sufficient with more than 5 years and the impact of cGVHD as well as pre-transplant

factors on long-term survival Nutlin-3a price were analyzed exclusively for subjects with active leukemia. Conclusion These data show that allo-HCT has the potential to cure active leukemia possibly via cGVHD, particularly in patients with favorable factors even when in non-remission. Further research is warranted to explore the essential factors contributing to the success of allo-HCT such as intensity of conditioning, and GVL effects mediated through cGVHD. Acknowledgements This work was supported by a Grant-in-Aid for Scientific Research from the Japanese Ministry of Education, Science, Sports, and Culture, and a grant from the Japanese Ministry of Health, Welfare, and Labour. References 1. Champlin Crenolanib order R, Gale RP: Acute myelogenous leukemia: recent advances in therapy. Blood 1987, 69:1551–1562.PubMed 2. Biggs

JC, Horowitz MM, Gale RP, Ash RC, Atkinson K, Helbig W, Jacobsen N, Phillips GL, Rimm AA, Ringdén O, et al.: Bone marrow transplants may cure patients with acute leukemia Paclitaxel supplier never achieving remission with chemotherapy. Blood 1992, 80:1090–1093.PubMed 3. Sierra J, Storer B, Hansen JA, Bjerke

JW, Martin PJ, Petersdorf EW, Appelbaum FR, Bryant E, Chauncey TR, Sale G, et al.: Transplantation of marrow cells from unrelated donors for treatment of high-risk acute leukemia: the effect of leukemic burden, donor HLA-matching, and marrow cell dose. Blood 1997, 89:4226–4235.PubMed 4. Greinix HT, Reiter E, Keil F, Fischer G, Lechner K, Dieckmann K, Leitner G, Schulenburg A, Hoecker P, Haas OA, et al.: Leukemia-free survival and mortality in patients with refractory or relapsed acute leukemia given marrow transplants from sibling and unrelated donors. Bone Marrow Transplant 1998, 21:673–678.PubMedCrossRef 5. Wong R, Shahjahan M, Wang X, Thall PF, De Lima M, Khouri I, Gajewski J, Alamo J, Couriel D, Andersson BS, et al.: Prognostic factors for outcomes of patients with refractory or relapsed acute myelogenous leukemia or myelodysplastic syndromes undergoing allogeneic progenitor cell transplantation. Biol Blood Marrow Transplant 2005, 11:108–114.PubMedCrossRef 6. Oyekunle AA, Kröger N, Zabelina T, Ayuk F, Schieder H, Renges H, Fehse N, Waschke O, Fehse B, Kabisch H, et al.: Allogeneic stem-cell transplantation in patients with refractory acute leukemia: a long-term follow-up. Bone Marrow Transplant 2006, 37:45–50.PubMed 7.

Mass spectrometry generated a list of 105 C. burnetii proteins in ACCM culture supernatants. Immunoblotting

of culture supernatants following growth of C. burnetii transformants expressing individual epitope-tagged versions of identified proteins confirmed secretion of 27 of these proteins. Secretion of epitope-tagged proteins also occurred during growth of C. burnetii in Vero host cells. An intact N-terminal signal sequence was required for secretion, indicating secreted proteins have a transient periplasmic location. Results Coxiella burnetii proteins are present in growth medium supernatant The Dot/Icm type IVB secretion system BIX 1294 of C. burnetii has been extensively studied [9, 10, 39]. However, little is known about other secretion systems of C. burnetii that are presumably important for intracellular parasitism. To determine if C. burnetii secretes proteins during axenic growth, bacteria were cultivated in ACCM-2 without neopeptone to eliminate media proteins. Following 7 days of growth, supernatant was concentrated and analyzed

by SDS-PAGE and silver staining (Figure 1). Many proteins were detected, with the majority GDC-0449 having a molecular weight below 20 kDa. In a discovery experiment to generate a list of potentially secreted proteins to further investigate, SDS-PAGE was conducted again and proteins stained with Coomassie G-250 to allow analysis by microcapillary reverse-phase HPLC nano-electrospray tandem mass spectrometry (μLC/MS/MS). A list of 105 proteins was generated (Additional files 1 and 2) with functions assigned based on the annotated genome of the C. burnetii Nine Mile RSA493 reference strain [18]. Sixteen proteins were annotated as hypothetical exported proteins, which represents 36% of the total proteins with this annotation in the predicted C. burnetii proteome [18]. Twenty-nine proteins,

such as translation initiation factor 1 (InfA) and ribosomal protein subunit L31P (RpmE), were predicted as cytoplasmic using the PSORTb v3.0.2 bacterial protein subcellular localization prediction program [40]. This result could be explained by a small amount of bacterial lysis releasing abundant cytoplasmic proteins that are then detected by highly sensitive mass spectrometry. The only Dot/Icm type IVB secretion Bay 11-7085 system substrate detected was CBU0937 [39]. However, type IVB-dependent secretion of CBU0937 was demonstrated using L. pneumophila as a surrogate host, and the protein contains a predicted signal sequence, which are typically not associated with Dot/Icm type IVB effectors [41]. Thus, CBU0937 may represent a false positive type IVB effector. Nonetheless, the lack of identified C. burnetii Dot/Icm type IVB secretion system substrates in culture supernatants indicates secretion via this mechanism requires host cell-derived signals. Figure 1 Multiple Coxiella burnetii proteins are present in growth medium supernatant. C.

Phytopathology 98:571–579PubMedCrossRef Gazis R, Rehner S, Chaverri P (2011) Species delimitation in fungal endophyte diversity studies and its implications in ecological and biogeographic inferences. Mol Ecol 20(14):3001–3013PubMedCrossRef Giménez-Jaime A, Aroca A, Raposo R, Garcia-Jiménez J, Armengol J (2006) Occurrence of fungal pathogens associated with

grapevine nurseries and the decline of young vines in Spain. J Phytopathol 154:598–602CrossRef Gonzáles V, Tello ML (2010) The endophytic mycota associated with Vitis vinifera in central Spain. Fungal Divers 47(1):29–42CrossRef Gramaje D, Armengol J (2011) Fungal trunk pathogens in the grapevine high throughput screening propagation process: potential inoculum sources, detection, identification, and management strategies. Plant Dis 95(9):1040–1055CrossRef Gramaje D, Garcia-Jiménez J, Armengol J (2010) Field evaluation of grapevine rootstocks inoculated with fungi associated with Petri disease and esca. Am J Enol Vitic 61(4):512–520CrossRef

Selleck SN-38 Graniti A, Surico G, Mugnai L (2000) Esca of grapevine: a disease complex or a complex of diseases? Phytopathol Mediterr 39:16–20 Green F III, Clausen CA (1999) Production of polygalacturonase and increase of longitudinal gas permeability in southern pine by brown-rot and white-rot fungi. Holzforschung 53(6):563–568CrossRef Green F III, Kuster TA, Highley TL (1996) Pectin degradation during colonization of wood by brown-rot fungi. Rec Res Devel Plant Pathol 1:83–93 Guo LD, Hyde KD, Liew ECY (2001) Detection and taxonomic placement of endophytic fungi within front tissues of Livistona chinensis based on rDNA sequences. Mol Phyl Evol 19:1–13CrossRef Halleen F, Crous PW, Petrini O (2003) Fungi associated with healthy grapevine cuttings in nurseries, with special reference to pathogens involved in the decline of young vines. Aust

Plant Pathol 32:47–52CrossRef Halleen F, Fourie PH, Crous P (2006) A review of black foot disease of grapevine. Phytopathol Mediterr 45:S55–S67 Higgins KL, Coley PD, Kursar TA, Arnold AE (2011) Culturing and direct PCR suggest prevalent host generalism among fungal endophytes of Methamphetamine tropical grasses. Mycologia 103(2):247–260PubMedCrossRef Hyde KD, Soytong K (2008) The fungal endophyte dilemma. Fungal Divers 33:163–173 International Organisation of Vine and Wine (2011). State of the vitiviniculture world market. OIV annual report, March. Available: http://​www.​indianwineacadem​y.​com/​2011_​note_​conj_​mars_​EN.​pdf. Accessed 8 March 2012. Ko Ko TW, McKenzie EHC, Bahkali AH, To-anun C, Chukeatirote E, Promputtha I, Abd-Elsalam KA, Soytong K, Wulandari NF, Sanoamuang N, Jonglaekha N, Kodsueb R, Cheewangkoon R, Wikee S, Chamyuang S, Hyde KD (2011) The need for re-inventory of Thai phytopathogens. Chiang Mai J Sci 38(4):1–13 Kuntzmann P, Villaume S, Larignon P, Bertsch C (2010) Esca, BDA and eutypiosis: foliar symptoms, trunk lesions and fungi observed in diseased vinestocks in two vineyards in Alsace.

The TFFBR also contains a pump, by which the water flow rate can be controlled. The main advantages of this TFFBR are (i) its high optical efficiency, (ii) it’s simple construction

method and (iii) the low investment costs involved in development. Further advantages are that oxygen transfers effectively into the water film and there is no need for TiO2 separation from the treated water, in contrast to reactors based on TiO2 slurries. An understanding of the mechanism of microbial photoinactivation during solar photocatalysis comes mostly from studies of bacteria [5, 7, 21]. The most common photocatalytic inactivation mechanism described is based on inactivation due to hydroxyl radicals and other reactive oxygen species (ROS) when bacteria come in contact with a solar-excited photosensitiser. GM6001 research buy This photooxidation process click here causes cell membrane disruption and increase cellular permeability,

with significant cell damage that eventually results in complete inactivation of the bacteria [13]. The conventional approach to assessing the viability of bacteria during solar disinfection is to enumerate samples after exposure to sunlight, using conventional plate counts on a suitable agar-based growth medium with incubation of plates in standard aerobic conditions (e.g. 24 h incubation at a suitable temperature). However, recent studies have demonstrated that reactive oxygen species (ROS), derived mainly from aerobic respiration during the enumeration process, may inactivate sub-lethally Sclareol damaged bacteria and prevent their growth and enumeration under aerobic conditions [22]. Such injured cells can only be cultured and counted under conditions where reactive oxygen species are neutralised (ROS-neutralised conditions) e.g. by supplementing the growth medium with the peroxide scavenger sodium pyruvate and incubating under anaerobic conditions

to prevent cellular respiration, allowing the bacteria to grow by fermentation [22–24]. This approach was taken in the present study; uninjured bacteria were enumerated under aerobic conditions while uninjured plus injured (ROS-sensitive) bacteria were enumerated under ROS-neutralised conditions, with the difference between the counts under both sets of conditions representing the number of injured bacteria in the sample. Even though bacteria have received more attention than other groups of microbes in solar photocatalysis research, bacterial pathogens of fish have been largely ignored in these studies, prompting the study reported here. Aeromonas hydrophila is a Gram-negative bacterium, known to be a primary fish pathogen [25]. A. hydrophila tends to be virulent towards most cultured and wild freshwater fish, especially trout, salmon, carp, catfish and tilapia. Red fin diseases and haemorrhagic septicaemia are mainly associated with A. hydrophila [26]. Antibiotics and several vaccines have been used to treat these infections, but extensive use of antibacterial agents has caused A.

In this case, the shape of Fe clusters is controlled by the thermodynamic stability of the planes in growth. But if the growth is controlled by the growing rate of crystal planes, the morphology of particles are check details changed depending on the condition [18]. As mentioned

above, three Si ad-atoms are remained in each half unit cell on the Si(111)-7 × 7-C2H5OH surface, and the deposited Fe atom may be stabilized by the dangling bond with one electron. In fact, the single-Fe atoms are recognized on the surface at low Fe coverage as shown in Figure 2a. If the Fe atoms are increased in a half unit cell, some kinds of interaction between the Fe atoms stabilize a cluster such as a pentagonal-base pyramid structure observed in the insert of Figure 2a. It should be reminded that the internal bond of Fe clusters may be stronger than selleck compound the interaction of Fe cluster with the surface, so that small Fe clusters grow instead of the Fe layers [19]. Figure 4 shows the simplified periodic grid of clusters in Figure 2d and the sizes of clusters. From Figure 4, it is known that the linearly arrayed Fe clusters take a size of about 5.4 × 4.7 nm, which

is much smaller than the reported critical size of Fe single magnetic domain clusters (~101 nm) prepared by the chemical methods. The 5-nm Fe clusters formed on Si(111)-7 × 7-C2H5OH showed unusual one-dimensional self-assembly with a regular periodic arrangement as shown in Figure 2c,d, which indicate some kinds of attractive interaction of large Fe clusters along the strings. This fact suggests the possibility for the preparation ID-8 of ca. 5-nm-size single magnetic domain Fe clusters. It is worthy of note that the straightly

linked chain structures appears on larger Fe clusters, just as shown in Figure 2c,d, and the authors presumed the formation of single magnetic domain of 5-nm Fe clusters. In addition, if we could oxidize and/or azotize the 5-nm Fe cluster, we could prepare the strong magnetic materials of FeO x and/or FeN x with single magnetic domain. Figure 4 The size of Fe clusters with 4 ML. In general, the periodical arrangement of Si atoms on Si(111)-7 × 7-reconstructed surface could result in the periodical surface potential field [20–22]. Then, the periodical surface potential field could restrain the growth of Fe cluster with certain periodicity. Based on the dimer-ad-atom-stacking (DAS) model of Si(111)-7 × 7-reconstructed surface [23], the side length of unit cell was 2.668 nm, just as shown in Figure 5a and the rhombus A in Figure 5b. According to the periodicity of rhombus unit cell in DAS model, the smallest rectangle structure with periodicity could be designed as the rectangle B-E shown in Figure 5b. Through the simple calculation, the width and length of the designed rectangle was 4.66 and 5.376 nm, respectively, which was corresponded well with the values of Fe clusters in Figures 2d and 4.

Important in this context is the observation that, after disregarding nonangiogenic subsets of NSCLC (which tend to obscure the association Foretinib research buy of Oct-4 with tumor angiogenesis), a subset of NSCLC tumors does not induce

angiogenesis, but instead co-opts the normal vasculature for further growth. On the basis of the previous finding that Oct-4 may be a major contributor to the maintenance of self-renewal in embryonic stem cells, we investigated the association of Oct-4 expression with self-renewal of NSCLC cells. The immunohistochemical analyses presented here showed clear Oct-4 staining in most sections, and RT-PCR showed Oct-4 mRNA in all NSCLC cell lines. Our data extend the previous report of Oct-4 overexpression in lung adenocarcinoma [20], providing the first demonstration that Oct-4 is also present in lung squamous cell carcinoma specimens, exhibiting an apparent difference in the degree of expression among sections analyzed. One possible explanation for these findings is that the genesis of lung buy Salubrinal adenocarcinoma and squamous cell carcinoma may be different. The former arises from mucous glands or the cells of bronchoalveolar duct junction and the latter grows most commonly in or around major bronchi. Further studies designed

to address the relationship between Oct-4 expression in endothelial precursors and the sites of origin of adenocarcinoma and squamous cell carcinoma are required to confirm this. Our data also showed that the degree of immunohistochemical staining was positively

correlated with poor differentiation of tumor cells and Ki-67 expression; this latter marker provides an opportunity to analyze the proliferative cell fraction in preserved tumor specimens. High levels of Oct-4 have been shown to increase the malignant potential of tumors, whereas inactivation of Oct-4 induces a regression of the malignant component [22]; moreover, knockdown of Oct-4 expression in lung cancer cells has been shown to facilitate differentiation of CD133-positive cells into CD133-negative cells [23]. These findings, taken together with our data, indicate that overexpression of Oct-4 in NSCLC tissues may maintain the second poorly differentiated state by contributing to tumor cell proliferation. On the other hand, down-regulation of Oct-4 expression has been shown to induce apoptosis of tumor-initiating-cell-like cells through an Oct-4/Tcl1/Akt1 pathway, implying that Oct-4 might maintain the survival of tumor-initiating cells, at least in part, by inhibiting apoptosis [13]. Whether an Oct-4-dependent pathway modulates apoptosis in clinical NSCLC samples or NSCLC cell lines has not yet been tested. Previous reports have indicated that tumor-induced angiogenesis is important in maintaining the poorly differentiated state and promoting metastasis in NSCLC [23, 24].

Nine replicates for each treatment.

Table 1 Metabolites with significant differences between mycelia of A. flavus grown in media with or without D-glucal eFT-508 supplier (40 mg/mL) Compoundsa Relative peak areab Fold increasec P valued Control D-glucal     Organic acids         Furanacetic acid 0.0184 ± 0.0039 2.9291 ± 0.2771 159.10 <0.01 Kojic acid 0.0942 ± 0.0333 0.2076 ± 0.0293 2.20 <0.01 Sugar metabolism         Ribitol 0.0066 ± 0.0038 0.0168 ± 0.0051 2.56 <0.01 Glycerol 0.0219 ± 0.0055 0.0514 ± 0.0350 2.34 <0.01 D-glucose 0.0133 ± 0.0060 0.1233 ± 0.0400 9.27 <0.01 D-galactose 0.0317 ± 0.0096 0.1750 ± 0.0743 5.53 <0.01 TCA intermediates         Succinic acid 0.0053 ± 0.0016 0.0020 ± 0.0005 0.37 <0.01 Malic acid 0.0023 ± 0.0013 ND ND ND Fumaric acid 0.0003 ± 0.0001

0.0002 ± 0.0000 0.53 <0.01 Fatty acids         Palmitic acid 0.1428 ± 0.0116 0.0856 ± 0.0144 0.60 <0.01 Stearic acid 0.0702 ± 0.0150 0.0468 ± 0.0072 0.66 <0.01 Oleic acid 0.1957 ± 0.0159 0.0377 ± 0.0093 0.19 <0.01 Linoleic acid 0.2647 ± 0.0219 0.1281 ± 0.0212 0.48 <0.01 Others         Glycine 0.0010 ± 0.0004 0.0004 ± 0.0002 0.39 <0.01 Pyrimidine 0.0018 ± 0.0005 0.0009 ± 0.0001 A 769662 0.53 <0.01 aIndividual metabolites were identified by GC-TOF MS, as described in the Methods. bRelative peak area as normalized to the peak area of heptadecanoic acid. cFolds represents the relative peak areas in D-glucal-treated samples/peak areas in the control. dStatistical differences between check details control and D-glucal treated samples were calculated by two-tailed Student’s t-test. N.D.: not detected. We next cultured A. flavus A 3.2890 in GMS

media with or without 40 mg/mL D-glucal, and measured kojic acid contents in media using a colorimetric method [19]. During the 5-d culture period the kojic acid contents in media with D-glucal were always higher (about 4 to 5 folds) than the control (Figure 4A). We also measured glucose content in the media and observed that, in the presence of D-glucal, the glucose content on the 4th and the 5th d were about 30% higher than those in the control media lacking D-glucal, suggesting that exogenous D-glucal inhibited the consumption of glucose (Figure 4B). Figure 4 Effects of D-glucal on kojic acid production, glucose consumption and NOR accumulation. (A) Production of kojic acid by A. flavus grown in GMS media with or without 40 mg/mL D-glucal. Values are presented as means ± S.D. (n = 3), from three independent experiments. (B) Glucose contents in media at different time points when cultured in the presence of 40 mg/mL D-glucal. (C) D-glucal inhibited NOR accumulation. The amount of NOR in GMS medium lacking D-glucal was set to 1, those in other samples were calculated accordingly. Values are presented as means ± S.D. (n = 3). D-glucal inhibited NOR production We used the A. flavus strain Papa 827 to decipher at which step D-glucal inhibits AF biosynthesis.