Introduction 1 2 4 1 2A 2B 3 2 4 5 5 6 i 5 6 7 8 9 3 5 7 10 10 3 7 3 6 3 i 11 max 2+ 11 3 3 12 50 12 7 5 12 7 12 7 50 12 10 12 7 12 7 10 3 12 3 3 3 Materials and methods Chemicals 3 Cell culture 2 Membrane preparations 7 13 14 7 13 15 3 7 2+ 2+ i 3 4 \documentclass[12pt]{minimal}
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\begin{document}$$ {\text{OD}}_{{600}}  = 1\;{\text{equals}}\;3 \times 10^{8} \;{{\text{cells}}} \mathord{\left/  {\vphantom {{{\text{cells}}} {{\text{ml}}}}} \right.  \kern-\nulldelimiterspace} {{\text{ml}}} $$\end{document} 16 Data analysis 50 Isolation of microorganisms from contaminated buffer solutions and classification  Bergey’s Manual of Systematic Bacteriology 17 Results 3 3 1 3 1 Fig. 1 3 a b c 3 n  Microbial contaminations in buffer solutions 3 3 3 3 3 6 3 1 18 3 3 m 2A 19 3 R H 3 20 12 3 21 2 3 3 3 3 3 3 3 3 Fig. 2 3 3  Isolation of microbial buffer contaminants 3 3 Fig. 3 3 a b 50 a \documentclass[12pt]{minimal}
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\begin{document}$${\text{rat}}\,{\text{cortex}} = 57.0 \pm 4.4\;{\text{nM}}{\left( {n = 3} \right)}$$\end{document} \documentclass[12pt]{minimal}
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\begin{document}$${\text{contaminant}}\;1 = 4.59 \pm 0.49\;{\text{nM}}{\left( {n = 3} \right)}$$\end{document} \documentclass[12pt]{minimal}
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\begin{document}$${\text{contaminant}}\;2 = 1,010 \pm 240\;{\text{nM}}{\left( {n = 2} \right)}$$\end{document} n 50 b \documentclass[12pt]{minimal}
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\begin{document}$$Achromobacter\,xylosoxidans = 13.8 \pm 2.7\;{\text{nM}}{\left( {n = 8} \right)}$$\end{document} \documentclass[12pt]{minimal}
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\begin{document}$$Achromobacter\,denitrificans = 253 \pm 46\;{\text{nM}}{\left( {n = 5} \right)}$$\end{document} \documentclass[12pt]{minimal}
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\begin{document}$$Acinetobacter\,lwoffii = 299 \pm 129\;{\text{nM}}{\left( {n = 5} \right)}$$\end{document}  \documentclass[12pt]{minimal}
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\begin{document}$$ {\text{IC}}_{{50}}  = 4.59 \pm 0.49\;{\text{nM}}\;{\text{vs}}{\text{.}}\;10\;{\text{nM}}\;{\left[ {^{3} {\text{H}}} \right]}{\text{adenine}} $$\end{document}  n \documentclass[12pt]{minimal}
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\begin{document}$$ {\text{IC}}_{{50}}  = 1008 \pm 240\;{\text{nM}} $$\end{document}  n 50  n \documentclass[12pt]{minimal}
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\begin{document}$$ {\text{IC}}_{{50}}  = 57.0 \pm 4.4\;{\text{nM}} $$\end{document} 3 Identification of microbial contaminants 1 Pseudomonas  Achromobacter Achromobacter  Acinetobacter  17 Acinetobacter A. lwoffii A. xylosoxidans A. denitrificans 1 Achromobacter Achromobacter 1 1 Table 1 Summary of classification test results using standard procedures and kits Test Contaminant 1 Contaminant 2 Contaminant 3 Morphology of colonies   Form Round Round Round   Diameter 1–2 mm 1 mm 1 mm   Color Grey Beige White   Brim Smooth Smooth Smooth   Surface Shiny Shiny Shiny   Cross section Raised Raised Raised   Gram stain Negative Negative Negative Biochemistry   Catalase + + +   Oxidase + + –   O/F test –/– –/– –/–   Motility + + –   Lysine/BBL Ox/Ferm Tube II – + n.d.   Result Achromobacter  Pseudomonas  Achromobacter  Acinetobacter  Api 20 NE 3 + + –   GLU, glucose + – –   GNT, gluconate + + –   CAP, caprate + – +   ADI, adipate + + –   MLT, maltose + + +   CIT, citrate + + –   PAC, phenylacetate + + +   OX, oxidase + + –   Result A. xylosoxidans A. denitrificans A. lwoffii VITEK 2   N-LGGH, γ-L-glutamyl (nutrient assimilation, NA) + + n.d.   P-BPHO, bis(p-nitrophenyl)phosphate + – n.d.   C-CIT, citrate – + n.d.   C-KGA, α-acetoglutaryc acid + + n.d.   N-LPROT, L-proline-NA + – n.d.   Result Achromobacter  Achromobacter  n.d.  n.d.  3 3 50 A. xylosoxidans \documentclass[12pt]{minimal}
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\begin{document}$$ {\text{IC}}_{{50}}  = 13.8 \pm 2.7\;{\text{nM}} $$\end{document} \documentclass[12pt]{minimal}
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\begin{document}$$ {\text{IC}}_{{50}}  = 4.59 \pm 0.49 $$\end{document} A. denitrificans A. lwoffii 50 D max A. xylosoxidans D max n D max n Structure–activity relationships A. xylosoxidans 7 4 \documentclass[12pt]{minimal}
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\begin{document}$$ {\text{IC}}_{{50}}  = 13.8 \pm 2.7\;{\text{nM}} $$\end{document} \documentclass[12pt]{minimal}
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\begin{document}$$ {\text{IC}}_{{50}}  = 59.1 \pm 19.6\;{\text{nM}} $$\end{document} \documentclass[12pt]{minimal}
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\begin{document}$$ {\text{IC}}_{{50}}  = 32,100 \pm 3000\;{\text{nM}} $$\end{document} 2 A. xylosoxidans i 3 50 i 7 50 A. xylosoxidans Fig. 4 3 Achromobacter xylosoxidans \documentclass[12pt]{minimal}
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\begin{document}$${\text{IC}}_{{50}} \,{\text{values}}:\,{\text{adenine}} = 13.8 \pm 2.7\;{\text{nM}}\;{\left( {n = 8} \right)}$$\end{document} \documentclass[12pt]{minimal}
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\begin{document}$${\text{hypoxanthine}} = 59.1 \pm 19.6\;{\text{nM}}\;{\left( {n = 3} \right)}$$\end{document} \documentclass[12pt]{minimal}
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\begin{document}$$2 - {\text{fluoroadenine}} = 32,100 \pm 3000\;{\text{nM}}\;{\left( {n = 3} \right)}$$\end{document}  Table 2 Comparison of affinities of adenine and selected compounds for the rat adenine receptor and bacterial binding sites determined in radioligand binding studies Compound Achromobacter xylosoxidans Rat brain cortical membranes 50 a i a Adenine b c 2-Fluoroadenine 32.1 ± 3.0 c 2-Hydroxyadenine d 29.4 ± 6.4 2,6-Diaminopurine d 4.95 ± 0.75 6 d 34.4 ± 8.1 Hypoxanthine 0.0591 ± 0.0196 45.0 ± 19.4 Uracil ≫10 (6 ± 1) ≫100 (−6 ± 2) Xanthine >10 (17 ± 6) >100 (23 ± 7) Uridine d ≫100 (−9 ± 3) Adenosine d c a b c i 7 d Discussion 3 6 7 5 10 3 11 12 3 11 12 12 3 3 12 3 1 3 1 3 3 3 3 6 3 3 12 3 11 12 3 12 3 12 3 3 3 Saccharomyces cerevisiae 3 Achromobacter Acinetobacter 17 A. xylosoxidans A. lwoffii 22 32 17 24 26 33 36 A. lwoffii Achromobacter  50 A. xylosoxidans A. xylosoxidans 7 5 6 A. lwoffii A. denitrificans A. xylosoxidans 3 3 A. xylosoxidans 37 Escherichia coli Bacillus Aspergillus nidulans Neurospora crassa 37 39 40 41 E. coli 42 3 A. xylosoxidans 43 44 Trypanosoma brucei brucei Crithidia luciliae 44 45 E. coli B. subtilis 42 46 47 46 47 3 Achromobacter Acinetobacter 42 47 A. xylosoxidans Achromobacter 3 11 12 12 10 12 7 12 3 A. xylosoxidans 37 48 37 48 52 46 53 C. luciliae 45 E. coli 53 B. subtilis 46 A. baumannii Pseudomonas  3 Achromobacter  Conclusions 3 3 A. lwoffii A. xylosoxidans, A. denitrificans 3