All of the chemicals used in this investigation were of analytical grade. Danofloxacin (DAN), ofloxacin (OFL), levofloxacin (LEV), garenoxacin (GAR), pefloxacin (PEF), gatifloxacin (GAT), clinafloxacin (CLI), sarafloxacin (SAR), lomefloxacin (LOM), tosufloxacin (TOZ), sparfloxacin (SPA), difloxacin (DIF), pazufloxacin (PAZ), marbofloxacin (MAR), moxifloxacin (MOX), rufloxacin (RUF), norfloxacin (NOR), ciprofloxacin (CIP), enrofloxacin (ENR), pipemidic acid (PIP), nalidixic acid (NAL), oxolinic acid (OXO), orbifloxacin (ORB), enoxacin (ENO), nadifloxacin (NAD), flumequine (FLU), bovine serum albumin (BSA), ovalbumine (OVA), casein, 1-ethyl-3-(dimethylaminopropyl) carbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), N,N-dimethylformamide (DMF), ethylenediamine hydrochloride, triethylamine, sodium borohydride, glutardialdehyde, 4-aminomethylfluorescein (4-AMF), 3,3′,5,5′-tetramethylbenzidine (TMB), and Tween-20 were Sigma-Aldrich (St. Louis, MO, USA) products. Complete and incomplete Freund’s adjuvants were produced by Becton Dickinson (Franklin Lakes, NJ, USA). Peroxidase-labeled anti-rabbit immunoglobulins were from the Gamaleya Institute of Microbiology and Epidemiology (Moscow, Russia). All other chemicals (salts and solvents of analytical grade) were from Khimmed (Moscow, Russia).
Sparfloxacin
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom
Sparfloxacin is a fluoroquinolone antibiotic used in the laboratory. It functions as an antibacterial agent that inhibits the activity of DNA gyrase and topoisomerase IV, essential enzymes for bacterial DNA replication and transcription.
Automatically generated - may contain errors
Lab products found in correlation
Ciprofloxacin, by Merck Group (6 mentions)
Norfloxacin, by Merck Group (5 mentions)
Ofloxacin, by Merck Group (4 mentions)
Moxifloxacin, by Merck Group (4 mentions)
Enrofloxacin, by Merck Group (3 mentions)
Levofloxacin, by Merck Group (3 mentions)
Nalidixic acid, by Merck Group (3 mentions)
Lomefloxacin, by Merck Group (2 mentions)
Difloxacin, by Merck Group (2 mentions)
Enoxacin, by Merck Group (2 mentions)
Gatifloxacin, by Merck Group (2 mentions)
Danofloxacin, by Merck Group (1 mentions)
Pefloxacin, by Merck Group (1 mentions)
Sarafloxacin, by Merck Group (1 mentions)
Tosufloxacin, by Merck Group (1 mentions)
Marbofloxacin, by Merck Group (1 mentions)
Pipemidic acid, by Merck Group (1 mentions)
Oxolinic acid, by Merck Group (1 mentions)
Orbifloxacin, by Merck Group (1 mentions)
Nadifloxacin, by Merck Group (1 mentions)
16 protocols using sparfloxacin
1
Development of Fluoroquinolone Immunoassays
2
Synthesis of Sparfloxacin-TCNE Charge Transfer Complex
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All the chemicals used were of analytical grade. The candidate donor sparfloxacin (SFX, 98.0%, HPLC, Batch# BCBN3519V), acceptor tetracyanoethylene (TCNE, 98%, Batch# S84306), SiO2 as silica gel (99%, Batch# 11615CH) and TiO2 (99.9%, Batch# 1317-70-0) were purchased from Sigma-Aldrich, USA. Cellulose (Batch# 33820) was purchased from Riedel-de Haen, USA. Acetonitrile (98%, GLC, BDH, 2 Batch# 9220, USA) and methanol (99.8%, Merck, Batch# I838218633, USA) were used as received. All the materials used were with a stated purity of >98% so were used without further purification.
3
Characterization of Multidrug-Resistant Klebsiella and Escherichia Strains
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EA27 is a multidrug resistant clinical isolate of Klebsiella aerogenes exhibiting a mutation in the quinolone resistance-determining region, a porin deficient phenotype and an energy-dependent efflux9 (link),25 (link). EA294 is the acrA::Kanr derivative strain of EA289 (not presented in this study), which itself is a kanamycin-sensitive derivative strain of EA2726 (link). EA3, EA117, KP 55, ARS100 and ARS108 are multidrug resistant clinical isolates of Klebsiella aerogenes (EA3, EA117) K. pneumoniae (KP55) and Escherichia coli (ARS100, ARS108) exhibiting a porin deficient phenotype and expression of the AcrB efflux pump27 (link),28 .
All strains were cultured in Mueller Hinton II broth (MHII) or Luria Broth (LB) at 37 °C. EA294 was grown in MHII or LB supplemented with kanamycin (50 µg.ml−1). The quinolones ciprofloxacin (CIP), enrofloxacin (ENR), fleroxacin (FLE), nalidixic acid (NAL), norfloxacin (NOR), sparfloxacin (SPA), chloramphenicol and erythromycin were purchased from Sigma-Aldrich. Rosoxacin (ROS) was purchased from LGC Standards Ltd. The quinolones were dissolved in Milli-Q water supplemented with 0.1 to 0.4% (v/v) NaOH, chloramphenicol and erythromycin were dissolved in 50% ethanol. Physicochemical parameters of the studied quinolones are listed in Supplementary Fig.1 .
All strains were cultured in Mueller Hinton II broth (MHII) or Luria Broth (LB) at 37 °C. EA294 was grown in MHII or LB supplemented with kanamycin (50 µg.ml−1). The quinolones ciprofloxacin (CIP), enrofloxacin (ENR), fleroxacin (FLE), nalidixic acid (NAL), norfloxacin (NOR), sparfloxacin (SPA), chloramphenicol and erythromycin were purchased from Sigma-Aldrich. Rosoxacin (ROS) was purchased from LGC Standards Ltd. The quinolones were dissolved in Milli-Q water supplemented with 0.1 to 0.4% (v/v) NaOH, chloramphenicol and erythromycin were dissolved in 50% ethanol. Physicochemical parameters of the studied quinolones are listed in Supplementary Fig.
4
Quantifying IL-17A+ T Cells by Flow Cytometry
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The differentiated T cells were incubated for 4 hours at 37°C in 5% CO2 with 0.1 mg/mL of monensin (Sigma, Saint Louis, USA), 1 μg/mL of ionomycin (Sigma, Saint Louis, USA), and 500 ng/mL of sparfloxacin (Sigma, Saint Louis, USA). Then, cells were stained with FITC-IL-17A monoclonal antibody (BD Bioscience, Franklin, USA) and APC-Cy™ 7-CD4 monoclonal antibody (BD Bioscience, Franklin, USA). The flow cytometry was performed in Cytek Athena system (Cytek, San Francisco, USA). FlowJo software was used to analyze the results.
5
Sparfloxacin-Impregnated Catheter Fabrication
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The samples were prepared by immobilization of sparfloxacin obtained from Sigma-Aldrich, on commercial latex catheters (Rusch, Germany). The catheters were cut into 0.5 cm long fragments and activated with 2% methanolic iodine solution for 1 h under gentle stirring at 40 °C. Next, the samples were removed, rinsed with distilled water, and dried. In the next stage, the samples were placed in Eppendorf tubes, immersed in 1 mL of 2 mg mL−1 solution of sparfloxacin and incubated in closed tubes for 12 h, at 60 °C with stirring at 100 rpm. Then the catheters were removed, rinsed with water, and left to dry overnight.
6
Photochemical Modulation of AhR Signaling
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VRCZ and VNO were purchased from Toronto Research Chemicals Inc. (North York, ON, Canada). They were dissolved in DMSO as 150 mM stock solutions and were used immediately or stored at −20 °C until used. HPLC-grade acetonitrile and methanol were purchased from Wako Pure Chemicals (Osaka, Japan). Photoproduct of VNO (P-VNO) was generated from VNO by irradiating with UVB irradiation (500 mJ/cm2). Afloqualone (AQ) was obtained from Tanabe Seiyaku Co., Osaka, Japan. The solution of PBS contained 0.3 mM AQ was prepared as described previously20 (link). Sparfloxacin, the AhR antagonists CH223191 and GNF351 were procured from Sigma-Aldrich, St. Louis, MO. KCZ, TBF and ITCZ were purchased from Wako Pure Chemicals, Osaka, Japan. Recombinant human TNF-α was purchased from R&D Systems, Minneapolis, MN.
7
Fluoroquinolone Compound Preparation
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The fluoroquinolones moxifloxacin (MXF), levofloxacin (LVX), norfloxacin (NOX), ofloxacin (OFX), sparfloxacin (SPX), and ciprofloxacin (CIP) were purchased from Sigma, UK. All compounds were made up in dimethyl sulfoxide (Sigma).
8
Antimicrobial Agents Acquisition and Preparation
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Ofloxacin, ciprOfloxacin, levOfloxacin, moxifloxacin, sparfloxacin, streptomycin, amikacin, kanamycin, capreomycin, and tobramycin were purchased from Sigma-Aldrich Co. (St Louis, MO, USA). All the antibiotics were freshly prepared before they were used.
9
Parabens and Dyes Analysis Protocol
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Methyl paraben, ethyl paraben, propyl paraben, butyl paraben, 5-(dimethylamino)naphthalene-1-sulfonyl chloride (Dansyl-Cl) trifluoroacetic acid (TFA), 4-dimethylaminoazobenzene-4′ sulfonyl chloride (Dabsyl-Cl), 1,3-benzothiazole-6-sulfonyl chloride (Batsyl-Cl), sparfloxacin, α-cyano 4-hydroxycinnamic acid (CHCA), sinapinic acid (SA), 2,5-dihydroxybenzoic acid (2,5-DHB), 2-mercaptobenzothiazole (2-MBT), 2-mercaptobenzoic acid (2-MBA) and 7-mercapto-4-methyl coumarin (7-MMC) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Acetonitrile, acetone, dimethyl sulfoxide (DMSO), sodium bicarbonate (NaHCO3), sodium carbonate, (Na2CO3), sodium hydroxide (NaOH), formic acid, trifluoroacetic acid, ethyl acetate, hexane and toluene were purchased from Merck (Darmstadt, Germany). Ethyl-d5 paraben (internal standard, IS) was obtained from Toronto (Toronto, Ontario, Canada). All reagents were of analytical grade. Deionized water was obtained using a Millipore Milli-Q (Bedford, MA, USA) water purification system.
10
Preparation of Fluoroquinolone Antibiotic Solutions
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Ciprofloxacin, enrofloxacin,
difloxacin, and sparfloxacin were purchased from Sigma-Aldrich (Gillingham,
U.K.). Pefloxacin, cinoxacin, and oxolinic acid were purchased from
Santa-Cruz Biotechnology (Dallas, TX, USA). Enoxacin was purchased
from TOKU-E (Washington, USA), and norfloxacin was purchased from
Merck Life Science UK Ltd. (Southampton, U.K.). Stock solutions of
antibiotics (100 mM) were prepared in DMSO, with the exception of
enoxacin which was prepared in 1 M NaOH. Stock solutions were stored
at −20 °C before use.
difloxacin, and sparfloxacin were purchased from Sigma-Aldrich (Gillingham,
U.K.). Pefloxacin, cinoxacin, and oxolinic acid were purchased from
Santa-Cruz Biotechnology (Dallas, TX, USA). Enoxacin was purchased
from TOKU-E (Washington, USA), and norfloxacin was purchased from
Merck Life Science UK Ltd. (Southampton, U.K.). Stock solutions of
antibiotics (100 mM) were prepared in DMSO, with the exception of
enoxacin which was prepared in 1 M NaOH. Stock solutions were stored
at −20 °C before use.
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