These potential insulin-induced epigenetic changes would functionally mimic both a (preceding) growth-promoting effect of an insulin-RB complex formation and a (subsequent) gene mutation pattern that may arise during the further evolution of these cells/tissues towards malignancy. In other words, the viral oncoprotein-like insulin Niraparib molecule [27, 31] would display two distinct properties that are functionally equivalent in terms of driving oncogenesis [18]. Moreover, the immunohistochemical identification of insulin in lung cancer tissue samples (whereby, besides the actual tumor cells, some normal pneumocytes were also revealed to be insulin-positive)

in buy Saracatinib the absence of detectable insulin PF299 mouse transcripts [32] additionally strengthens the concept of a pathological spread of (blood-borne) insulin in malignant diseases.

Beyond insulin, there are also other candidate molecules that could undergo an oncoprotein metastasis, e.g. osteopontin. Accordingly, it has been shown that osteopontin is found in premalignant and malignant cells derived from patients with tumors of the oral cavity [33] and, moreover, that osteopontin translocates to the nuclei of mitotic cells [34]. Entirely consistent with the oncoprotein metastasis concept and intriguingly, it has furthermore been shown that primary tumor-derived and blood-borne osteopontin is able to promote the microenvironmental changes necessary for

distant metastatic seeds [35]. Most second recently, a known amino acid labeling technique has been extended to investigate intercellular communication via both secreted and internalized proteins such as metastasis associated protein 3 and retinoblastoma binding protein 7 [36]. It will therefore be interesting to probe in future studies as to whether these proteins can add to insulin and osteopontin as mediators of the proposed oncoprotein metastasis phenomenon. Since it thus appears that that there are various proteins that cross subcellular borders and thereby contribute to carcinogenesis, a therapeutic strategy that suggests itself in order to counteract these microbial infection-like, transcellular processes of malignancy would be to administer cell-permeable agents that directly block these mobile oncoproteins. Possible pharmacological candidates for such intervention are cell-penetrating tumor suppressor peptides, in particular those targeting the RB and nucleocrine pathways [17, 18, 28, 30, 37–40]. In this context, a parallel is noteworthy: in the same way as insulin’s internalization into cells is not saturable [41] nor is that of a 16-amino acid fragment derived from the Antennapedia homeodomain and termed “”Penetratin”" either [42].

Mol

Microbiol 2000,36(3):585–593.CrossRefPubMed 8. Chen CJ, Elkins C, Sparling PF: Phase variation of hemoglobin utilization in Neisseria gonorrhoeae. Infect Immun 1998,66(3):987–993.PubMed 9. Jordan PW, Snyder LA, Saunders NJ: Strain-specific differences in Neisseria gonorrhoeae associated with the phase variable gene repertoire. BMC VX-770 chemical structure Microbiol 2005,5(1):21.CrossRefPubMed 10. Richardson AR, Stojiljkovic I: Mismatch repair and the regulation of phase variation in Neisseria meningitidis. Mol Microbiol 2001,40(3):645–655.CrossRefPubMed 11. Kline KA, Sechman EV, Skaar EP, Seifert HS: Recombination, repair and replication in the pathogenic Neisseriae: the 3 R’s of molecular genetics of two click here human-specific bacterial pathogens. Mol Microbiol 2003,50(1):3–13.CrossRefPubMed 12. Skaar EP, Lazio MP, Seifert HS: Roles of the recJ and recN genes in

homologous recombination and DNA repair pathways of Neisseria gonorrhoeae. J Fedratinib Bacteriol 2002,184(4):919–927.CrossRefPubMed 13. Campbell LA, Yasbin RE: A DNA excision repair system for Neisseria gonorrhoeae. Mol Gen Genet 1984,193(3):561–563.CrossRefPubMed 14. Nyaga SG, Lloyd RS: Two glycosylase/abasic lyases from Neisseria mucosa that initiate DNA repair at sites of UV-induced photoproducts. J Biol Chem 2000,275(31):23569–23576.CrossRefPubMed 15. Campbell LA, Yasbin RE: Deoxyribonucleic acid repair capacities of Neisseria gonorrhoeae : absence of photoreactivation. J Bacteriol 1979, 140:1109–1111.PubMed 16. Campbell LA, Yasbin RE: Mutagenesis of Neisseria gonorrhoeae: Absence of error-prone repair. J Bacteriol 1984, 160:288–293.PubMed 17. Ambur OH, Davidsen T, Frye SA, Balasingham SV, Lagesen K, Rognes T, Tonjum T: Genome dynamics in major bacterial pathogens. FEMS Microbiology Reviews 2009,33(3):453–470.CrossRefPubMed 18. Bryant DW, McCalla DR, Leeksma Astemizole M, Laneuville P: Type I nitroreductases of Escherichia coli. Can

J Microbiol 1981,27(1):81–86.CrossRefPubMed 19. Jorgensen MA, Trend MA, Hazell SL, Mendz GL: Potential involvement of several nitroreductases in metronidazole resistance in Helicobacter pylori. Arch Biochem Biophys 2001,392(2):180–191.CrossRefPubMed 20. Koder RL, Haynes CA, Rodgers ME, Rodgers DW, Miller AF: Flavin thermodynamics explain the oxygen insensitivity of enteric nitroreductases. Biochemistry 2002,41(48):14197–14205.CrossRefPubMed 21. Watanabe M, Nishino T, Takio K, Sofuni T, Nohmi T: Purification and characterization of wild-type and mutant “”classical”" nitroreductases of Salmonella typhimurium . L33R mutation greatly diminishes binding of FMN to the nitroreductase of S. typhimurium. J Biol Chem 1998,273(37):23922–23928.CrossRefPubMed 22. Zenno S, Kobori T, Tanokura M, Saigo K: Purification and characterization of NfrA1, a Bacillus subtilis nitro/flavin reductase capable of interacting with the bacterial luciferase. Biosci Biotechnol Biochem 1998,62(10):1978–1987.CrossRefPubMed 23.

62 plastocyanin – ↓ LIC12829 (LA0790) gltA -1.53 citrate (Si)-synthase – - – carbohydrate transport and metabolism           (G)   -1.82 phosphonomutase – ↓ LIC12331 (LA1416) mgsA -1.72 methylglyoxal synthase – - LIC12733 (LA0909)   -1.58 adolase – ↓ LIC12233 (LA1532)           – amino acid transport and metabolism (E)   -2.17 dioxygenase superfamily selleckchem protein – - LIC10069 (LA0076) glnK -2.17

nitrogen regulatory protein PII – - LIC10440 (LA3807) csdB -1.60 selenocysteine lyase – - LIC20204 (LB267) speD -1.54 adenosylmethionine decarboxylase – - LIC20239 (LA-SPN3792) gltB -1.53 glutamate synthase (NADH) – - LIC12694 (LA0956)   -1.52 lactoylglutathione or related lyase – - LIC10460 (LA3782)           – nucleotide transport and metabolism (F)   -1.65 purine-nucleoside phosphorylase – - LIC13399 (LA4248) adk -1.55 adenylate kinase – - LIC12852 selleck (LA0760)           Transmembrane Transporters inhibitor – coenzyme transport and metabolism (H) ubiG -1.86 2-polyprenyl-3-methyl-5-     LIC10737 (LA3436)     hydroxy-6-metoxy-1,4- – -       benzoquinol methylase     – lipid transport and metabolism (I) ivd -1.77   – - LIC10363 (LA0414)     isovaleryl-CoA dehydrogenase     – inorganic ion transport and

metabolism amtB -3.10   – - (P) kdpA -2.09 ammonia permease ↑ – LIC10441 (LA3806)     potassium-transporting ATPase A     LIC10990 (LA3112)     chain     aGene ID is based on predicted ORFs of whole-genome sequence of L. interrogans serovar Copenhageni. Gene ID of corresponding serovar Lai is in parenthesis. ORFs of unknown or poorly characterized function were excluded from this table. bPrevious microarray data on the effect of overnight 37°C upshift [11] compared to growth at 30°C. cPrevious microarray data on the effect of osmolarity upshift [13] compared to EMJH medium. d ↑ represents up-regulation of gene expression and ↓ represents down-regulation of gene expression. Information storage and processing Putative transcriptional regulators

including Phospholipase D1 a protein in the PadR family (encoded by LIC10378) were up-regulated in response to serum. PadR has been shown to be a transcriptional repressor of padA gene (encoding a phenolic acid decarboxylase) expression in response to phenolic acid stress in Lactobacillus plantarum [46, 47]. However, the target of the leptospiral PadR homolog remains unknown. In the presence of serum, several subunits of 30S and 50S ribosomal proteins of Leptospira were repressed, possibly due to the shift of energy to produce other gene products that are needed for survival in serum. Reduction of ribosomal gene expression has also been found in organisms under various stress conditions such as Streptococcus pneumoniae isolated from infected blood [48], Campylobacter jejuni, Staphylococcus aureus, and Helicobacter pylori in response to acid shock [49–51], and E. coli under anaerobic and acidic conditions [52] and nitrogen and sulfur starvation [53].

The expression of tk and MCP-1 protein were detected by western blot 48 h after transfection. a: SKOV3/tk. b: SKOV3/MCP-1. c: SKOV3/neo. d: SKOV3/tk-MCP-1. RT-PCR Total RNA was extracted as described

previously and RT-PCR was performed comprising 33 thermal cycles of 95°C for 5 min, 94°C for 1 min, 58°C for 1 min, 72°C for 1 min and 72°C for 7 min. The same condition was used in MCP-1 amplification except 30 cycles in total. SNX-5422 Cell culture and retrovirus infection The human epithelial ovarian cancer cell line SKOV3 was used in vitro and vivo. SKOV3 cells were infected with supernatant of retrovirus at high titre containing pLXSN/tk-MCP-1(5.3 × 105 CFU/ml), pLXSN/tk(6.0 × 105 CFU/ml), pLXSN/MCP-1(4.8 × 105 CFU/ml) and pLXSN/neo(4.5 × 105 CFU/ml) at various volumes (100 μl, 200 μl, 500 μl or 1 ml), supplied with RPMI-1640 with 10% NBS to 2 ml, and then added polybrene (the concentration of polybrene at 8 μg/ml). Three hours later, cells were supplied with RPMI-1640 with 10% NBS to 8 ml and cultured for 2–3 days at 37°C in a 5% CO2 atmosphere. G418 at 600 μg/ml was added into 4 kinds

of cells. Ten days later, cells which survived in medium containing G418 at 600 μg/ml named SKOV3/tk-MCP-1, SKOV3/tk, SKOV3/MCP-1 and SKOV3/neo. Western blot Proteins were 3-Methyladenine solubility dmso extracted using protein extraction reagent, 48 h after transfection and save at −20°C, following a protocol provided by the manufacture. MCP-1 protein and tk protein expressions were detected with western blot. Proteins with equal amount were separated by appropriate concentration SDS-polyacrylamide gel electrophoresis and transferred onto PVDF membrane (Millipore, Billeriaca, AZD9291 cost MA, USA). The membranes were blocked in TBST for 1 h at room HSP inhibitor temperature and then incubated with primary antibodies fo tk (1:500,

Abcam, United Kingdom), MCP-1 (1:500, Santa Cruz Biotechnology) and β-actin (1:5000, Boston, MA) overnight at 4°C The membranes were then washed three times with TBST, followed by incubating with HRP-labeled secondary antibodies (KPL, Gaithersburg, MD, USA) (1:5000). Bound antibody was visualized using ECL detection reagent (Merck, Darmstadt, Germany). Antitumor effect of GCV The number of viable cells were determined by 3-(4, 5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide (MTT) assay. There were 4 experimental groups including SKOV3/tk, SKOV3/MCP-1, SKOV3/tk-MCP-1 and SKOV3/neo. Cells were re-suspended in fresh culture medium at the density of 2 × 104 cells/ml, 180 μl suspension were incubated in 96-well plates. The cells were treated with 20μl GCV at the concentrations of 10−2, 10−1, 1, 10, 102, 103 μg/ml for 72 h at 37°C in 5% CO2 incubator. SKOV3/tk-MCP-1 and SKOV3/neo seeded by same way was added GCV (1.0 μg/ml, 0.1 μg/ml) incubated for 24, 48, 72 and 96 h to detect time toxicity of GCV. 20μl Sodium Chloride was added to controls.

The cross-tabulation of all variables with the severity score regrouped in three categories is given in Appendix 5. Table 3 presents the odds ratios of the full

model including all the variables selected in the above step as well as the model which is the result of the backward selection with a 5 % p value for removal. All variables with several categories (e.g., age classes) were either removed or kept jointly. Table 3 EPZ-6438 order Ordinal logistic regression analyses CP-868596 manufacturer of predictors on the severity score   Full modela Selected modelb OR 95 % CI OR 95 % CI Gender  Male –        Female 2.20 0.73, 6.61     Age  <35          35–44 0.74 0.25, 2.17      45 and more 1.13 0.38, 3.39     Initial symptoms of psychological distress  None –   –    Minor 3.25 1.03, 3.43 3.02 0.99, 9.23  Moderate 4.80 1.40, 16.5 5.47 1.71, 17.5

 Severe 44.4 7.95–248 54.2 10.7, 275 Perception of the employer’s response  Adequate –        No employer 3.90 1.12, 13.5 3.73 1.09, 12.8  Inadequate 2.87 1.04, 7.94 2.86 1.06, 7.66 Previous experience of violence and jobs with high risk and awareness of violence  No/other jobs –   –    No/high risk and awareness of violence jobs 13.0 2.43, 69.9 11.0 2.08, 58.3  Yes/other jobs 0.54 0.18, 1.63 0.70 0.25, 1.97  Yes/high risk and awareness of violence jobs 0.72 0.22, 2.37 0.61 0.19, 1.90 aModel including jointly all factors which were statistically significant in simple regression GSI-IX in vitro analyses bModel obtained from the full model by backward selection The strongest feature of the regression analysis was that the severity score increased with the severity of the initial symptoms of psychological distress. On the other hand, age and sex were no longer found to be significant independent variables. The analysis of the interaction between previous experience of violence and “high risk and awareness of violence jobs” vs. “other jobs” (i.e., “moderate and low risk and awareness of violence jobs”) revealed notable results. First, in the “other jobs,” previous experience of violence did not affect severity of consequences

of the violent event. Second, in the “high BCKDHA risk and awareness of violence jobs,” the severity score was higher in the group without previous experience of violence. The significance of independent variables differed when considering their effect on the three components of the severity score taken separately (Table 4). For psychological consequences, the significant independent variables were initial symptoms of psychological distress and perceived lack of support from employer. For the consequences on work and employment, only severe initial symptoms of psychological distress were significant. For physical consequences of violence, only “no employer” (i.e., being an independent worker) was significant.

Acknowledgments This work was supported by the National Key Basic Research Program of China (2013CB922303, 2010CB833103), the National Natural Science Foundation of China (60976073, 11274201, 51231007), the 111 Project (B13029), the National Found for Fostering Talents of Basic Science (J1103212), and the Foundation for Outstanding Young Scientist in Shandong Province (BS2010CL036). References

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2011; Chu et al. 2011). The existence of taxane gene clusters in fungi and plants raises intriguing questions about the origin and evolution of these highly-specialized biosynthetic pathways, and the potential for HGT from fungi to Taxus trees. However, HGT between distantly-related organisms is a rare evolutionary event which is also constrained by the amount of genetic information transferred and genetic barriers

involving incompatible regulation and codon usage. This contrasts Regorafenib manufacturer sharply with the widespread observation of Taxol Selleck BI 10773 biosynthesis in many different endophytic fungi (Kurland et al. 2003). Material and methods Isolation of endophytic fungi from Taxus spp. plant material Endophytic fungi were isolated as previously described by Guo et al. (2006). Bark segments (0.5 × 0.5 cm) were removed with a sterile scalpel and surface sterilized for 5 min in 70 % ethanol. The inner bark was separated from the outer layer and placed on PDA agar (Carl Roth GmbH, Karlsruhe, Germany) supplemented with 25 mg/L streptomycin. The plates were incubated at room temperature until fungal growth was visible. The mycelium was then transferred to fresh plates using the hyphal tip method. Cultivation of endophytic fungi

The isolated endophytic fungi were cultivated on solid media, PDA (Carl Roth GmbH) supplemented with streptomycin or on YM-6.3 agar (0.4 % (w/v) glucose, 0.4 % (w/v) yeast extract, 2 % (w/v) malt extract, pH 6.3, 1.5 % (w/v) agar-agar). The fungi were transferred to fresh PF299804 manufacturer plates at weekly intervals by cutting out a piece of overgrown agar. In liquid culture, the fungi were grown in 0.6–10 L YM-6.3 medium (120 rpm in the dark) for 3 weeks or until no more glucose could be detected. The fungi were also cultivated in S7 medium as described for taxane-producing endophytes (Stierle et al. 1993). Taxoid extraction For taxane analysis, the Fenbendazole fungal culture media were extracted twice with an equal volume of chloroform. The organic phase was then dried over magnesium sulfate, evaporated to dryness and the residue was redissolved

in 3–5 mL methanol. Plant material (30 g Taxus needles or tobacco leaf tissue) was lyophilized and extracted with 1:1 dichloromethane/methanol in a Soxhlet extractor. The organic solution was evaporated to dryness and redissolved in dichloromethane. After two rounds of extraction with water, the organic layer was dried over magnesium sulfate, evaporated to dryness and the residue was redissolved in methanol (Witherup et al. 1990). Anti-taxane immunoassay (competitive inhibition enzyme immunoassay, CIEIA) The anti-taxane immunoassay was carried out according to the manufacturer’s instructions (Cardax Pharmaceuticals, Hawaii). A standard curve for taxane quantitation was made using Taxol concentrations of 111, 37, 12.33, 4.11, 1.37, 0.46 and 1.15 ng/mL (Table S1). The samples were analyzed using three dilutions.

Peptide conjugated to antibody has been used for delivery of siRNA to T cells of humanized mice to suppress HIV infection [35]. PEI polymers are able to successfully complex DNA molecules and they also have distinct transfection efficiency in a wide variety of cell types compared CBL0137 research buy to some other polymer systems described later. PEI derivatives cross-linked with different acrylates showed high gene expression in the lung or the spleen in mice. They also showed only little toxicity in cell culture experiments [36]. In vivo application of this polymer promises to take the polymer-based vector to the next level where it

can undergo clinical trials and then could be used for delivery of therapeutics in humans [37]. PLL is another cationic polymer, and its efficiency in gene delivery depends on its molecular weight. In low molecular weight, its complex with DNA is less soluble and rapidly removed by the Kupffer cells of the liver. With increasing the molecular weight, the efficiency of PLL is enhanced, interestingly [38]. Selleck Cilengitide Dendrimers are three-dimensional polymers with spherical, highly branched structures. Frequently used dendrimers are polyamines, polyamides, or polyesters. Because of its high transfection efficiency, polyamidoamine (PAMAM) is the most commonly used. The type of amine groups and the size of dendrimers have an influence

on their transfection efficiency. The primary amine groups promote DNA cellular uptake because of their participation in DNA binding but the buried tertiary amino groups act as a proto-sponge in endosomes and enhance the release of DNA into the cytoplasm. The studies show that with increasing the size and diameter, dendrimers enhance transfection efficiency [39, 40]. Recently, nitrogen-core

poly(propyl Mannose-binding protein-associated serine protease ether imine) (PETIM) dendrimer DNA complexes have been investigated and results showed low toxicities and efficient gene delivery vector properties. Quantitative estimation, using luciferase assay, showed that the gene transfection was at least 100 times higher when compared to poly(ethyleneimine) branched polymer, having similar number of cationic sites as the dendrimer [40]. Poly lactic-co-glycolic acid (PLGA)-based nanoparticles have been recognized as a potential vector to deliver genes. They are used in gene therapy for tumor and other miRNA-related diseases such as diabetes and cardiovascular and neurodegenerative diseases. The researches show that PLGA makes an improved safety profile in comparison with high-molecular weight PEIs and www.selleckchem.com/products/AZD2281(Olaparib).html liposome. Also, it is demonstrated that serum cannot inhibit the transfection activity of these nanoparticles [41]. PLGA nanoparticles are internalized in cells through pinocytosis (fluid phase) and also through clathrin-based endocytosis. These nanoparticles rapidly escape the endo-lysosomes and enter the cytoplasm within 10 min of incubation [24].

By definition, Wortmannin monoterpenes possess a carbon skeleton based on two C5 units originating from isopentenyl pyrophosphate (IPP), which is synthesized via the mevalonate (in eukaryotes) or the find more mevalonate-independent

pathway (in prokaryotes and plant plastids) [12–14]. Mainly, plant monoterpenes are produced via the latter pathway, but the metabolic cross linkage between both has been reported in several species [15, 16]. Monoterpenes are together with sesquiterpenes the major constituents of essential oils. Due to their status – they are generally recognized as safe (GRAS) [17] – and their odorous properties, these substances are widespread in the food, cosmetics, flavour and fragrance industry [18]. Monoterpenes are utilized as energy and carbon source by several aerobic microorganisms, a fact known since the 1960s [19–21]. Most reports dealt with Pseudomonas species, e.g. [22–28], but also Bacillus stearothermophilus[29], Rhodococcus erythropolis[30], and Enterobacter cowanii[31] metabolize these hydrocarbons. The microbial degradation of α-pinene and limonene, one of the most widespread monoterpenes in nature, involve complex and multiple pathways that comprise in large part oxidation reactions [30, JSH-23 datasheet 32–34]. In addition these studies revealed the importance of oxygenases, which

catalyze hydroxylation reactions with molecular oxygen as co-substrate [35–38]. Under anaerobic conditions, the biochemistry GNAT2 for the activation of these natural abundant alkenes seems to follow a totally different mechanism. The first evidence for the anaerobic degradation of monoterpenes were seven nitrate-reducing enrichment cultures with monoterpenes as sole carbon source [39]. Isolation

led to the description of four Alcaligenes defragrans strains, including strain 65Phen isolated with α-phellandrene [40]. A taxonomic study transferred these strains in the novel genus Castellaniella within the Alcaligenaceae, as C. defragrans[41]. The betaproteobacterium is capable of degrading a broad substrate range of a-, mono-, and bicyclic monoterpenes (Figure  1) [40]. Initial metabolite studies on the anaerobic monoterpene degradation pathway in C. defragrans elucidated the demand for a sp2-hybridized C1-atom as structural prerequisite for monoterpenes utilization [42] as well as the formation of geranic acid as intermediate [43], which is likely degraded on a modified β-oxidation pathway [44, 45]. These findings proposed the degradation of β-myrcene via hydration to linalool, followed by isomerisation to geraniol, and then two oxidations to geranial and to geranic acid [43]. The genes and proteins involved this pathway were recently identified [46, 47] (Figure  2).

Comparisons of CYC202 order a large collection of carbon sources reveal that sugars that are normally oxidized through the hexose monophosphate or glycolytic pathway

such as glucose, raffinose and mannose are efficient carbon sources for AF productions [23], while lactose and most amino acids excluding aspartate are considered to be unsuitable carbon sources for AF production [11, 26]. AFs are usually produced in parallel with fatty acid biosynthesis following the rapid growth and sugar utilization phase, as common precursors such as acetyl-CoA and malonyl-CoA derived from glucose catabolism are utilized in both pathways [18]. As many carbohydrates are able to induce AF production, Abdollahi and Buchanan (1981) believe that utilization of readily metabolized carbohydrates may result in elevated energy status which in turn induces AF biosynthesis [23]. Wiseman and Buchanan (1987) note that, although mycelia grow well in media with low concentrations of suitable sugars, AFs are produced only when sugar concentrations are higher Selleckchem Alvocidib than 0.1 M, and in which reduced mycelial growth and inhibited TCA cycle activity are observed [27]. Addition of TCA cycle intermediates inhibits AF production, suggesting that glucose may MK-2206 in vitro regulate AF productions

through inhibition of the TCA cycle [25, 26]. Recent studies have revealed cell density-dependent sclerotium formation and AF production in media with glucose and sorbitol as the carbohydrate sources, which is regulated through non-cell autonomous factors [28, 29]. In nature, seeds with high protein and lipid content, such as peanut and cotton, are more susceptible

to high AF production than starchy seeds like rice and sorghum [1]. It has also been shown in maize that mycelial growth and AF production occur primarily in the embryo and the aleurone layer where mainly storage proteins and lipids are accumulated [30, 31]. Removal of oil from ground cotton seeds greatly enhances AF production, Interleukin-2 receptor suggesting that lipids are not essential for optimal AF biosynthesis [32]. Fatty acids may stimulate or inhibit AF production through the presence of various oxidation-derived oxilipins [33–36]. The influence of protein and peptone on AF biosynthesis remains largely unknown. In this study we investigated how AF production by Aspergillus was influenced when peptone was used as the sole carbon source. Contrary to expectations, we observed spore density- and peptone concentration-dependent AF production with peptone as the sole carbon source. AFs were only produced in the PMS medium when initial spore densities were 104 spores/ml or lower. In contrast, mycelia cultured in the PMS medium with higher initial spore densities or with increased peptone concentrations grew rapidly but without AF production.