Endoxan

Additionally, 2–3-month-old female Lewis rats were treated with the immunosuppressant cyclophosphamide (Endoxan, Baxter, Prague, Czech Republic, 75 mg/kg i.p.) 5 days and 1 day before they were infected with A. fumigatus, to induce neutropenia. The animals repeatedly received (5 days, 1 day before, and on the day of inoculation) the antibiotic teicoplanin (Targocid, Sanofi-Aventis, Prague, Czech Republic, 35 mg/kg—5 days before i.m., or 25 mg/kg i.m.—1 day before and on the day of inoculation) to avoid bacterial superinfections, and additional antibiotics were administered by drinking water (Ciprofloxacin, Fresenius Kabi, Prague, Czech Republic, 2 mM, Colomycin, Teva, Prague, Czech Republic, 0.1 mM) for the duration of the experiment. Infection in the lung was established by intratracheal inoculation of 100 μL of A. fumigatus spores (10⁹ CFU/mL A. fumigatus ATCC 46645) using TELE PACK VET X LED system equipped with a flexible endoscope (Karl Stroz GmbH and Co. KG, Tuttlingen, Germany) [36 (link)].

Doxorubicin (ADRIBLASTINA® RD; Pfizer, New York, NY, USA) and cyclophosphamide (ENDOXAN®; Baxter Healthcare, Deerfield, IL, USA) solutions were administered on the same day via intraperitoneal injection at a dose of 2 mg/kg (doxorubicin) and 100 mg/kg (cyclophosphamide). To obtain a complete response for models HBCx-17 and HBCx-6, the same dose of AC chemotherapy was applied a second time, 3 weeks after the first injection. AC chemotherapy was applied to 68 mice of tumor graft model HBCx-17, 32 mice of HBCx-10, 35 mice of HBCx-6, and 30 mice of HBCx-14 model, not including the control group.

This experimental study was performed on 54 male Wistar rats (4 months old) weighing 200-250 gr. Animals were purchased from the Laboratory Animal Breeding Center of Baqiyatallah University of Medical Sciences (Iran) and were randomly divided into six groups (n = 9): group 1 (control) underwent the normal diet and water; group 2 (sham) received 2 ml/day normal saline; group 3 (positive control) received 300 mg/kg/day Ceratonia extract (14); group 4 (Ceratonia + CP) received Ceratonia extract (300 mg/kg/day) + 5 mg/kg/day CP (Endoxan, baxter oncology gmbh, Germany) after 4 hr; group 5 (CP) received 5 mg/kg/day CP (15, 16) + normal saline 4 hr after it; and group 6 (CP + Ceratonia) received Ceratonia extract (300 mg/kg/day) 4 hr after 5 mg/kg/day CP. CP and Ceratonia extract were administered for 28 days by gavage. Treatment period was selected on the basis of previous studies that demonstrated 5 mg/kg/day of CP treatment for 28 days induces toxicity in the testis tissue (15, 16).

BMC transplantation from C57BL/6 and C3HA donor mice or transgenic GFP-expressing C57BL/6 mice was performed similarly. mdx mice were irradiated on RAP-150/300-14 X-ray apparatus (Russia) at a dose of 2 or 3 Gy. One day after irradiation, the mice were injected with 0.2 mL (15–20 × 10⁶ nucleated BMCs) of BMC suspension in DPBS Ca⁻, Mg⁻ in the jugular vein.
After transplantation, the mice that were injected with BMCs from C3HA mice donors were additionally injected with the immunosuppressor Endoxan (cyclophosphamide; Baxter Oncology GmbH, Halle, Germany) either intraperitoneally or intramuscularly.

Mice received BU (10 mg kg⁻¹ day⁻¹ from days –7 to –4; Busilvex, Pierre Fabre Pharma, Germany) and CY (100 mg kg⁻¹ day⁻¹ from days −3 to −2; Endoxan, Baxter Healthcare, USA) i.p. (BU/CY). Day of stem cell injection was assigned as day 0 and the days before stem cell injection are numbered backwards.

GSK 269962 (N-[3-[[2-(4-Amino-1,2,5-oxadiazol-3-yl)-1-ethyl-1H-imidazo[4,5-c]pyridin-6-yl]oxy]phenyl]-4-[2-(4-orpholinyl)ethoxy]benzamide, Tocris), a potent ROCK inhibitor (IC50 values: 1.6 and 4 nM for ROCK1 and ROCK2, respectively) was dissolved in DMSO and administered intravenously (i.v.) at a daily dose of 30 mg/kg. CYP (Endoxan, Baxter Deutschland GmbH, Unterschleißheim, Germany) was diluted with saline (0.9% NaCl) and administered i.p. at a single dose of 200 mg/kg. The control rats received volume-matched saline i.p. The doses of the administered agents were chosen based on the literature data and were confirmed/adjusted in our laboratory in preliminary experiments (Hidalgo-Lucas et al. 2016 (link); Juszczak et al. 2010 (link); Kobayashi et al. 2009 (link); Wróbel and Rechberger 2015 (link); Wróbel and Rechberger 2016a (link); Wróbel and Rechberger 2016b (link); Wróbel and Rechberger 2016c (link)).

Teneral tsetse flies were fed 24–48 hours after emergence with a T.b.brucei AnTAR1, AnTat1.1E^DsRED or AnTat1.1E^TagGFP2 infected blood meal supplemented with 10 mM reduced L-glutathione to increase trypanosome infection establishment [31 (link)]. Parasitized blood was harvested with heparin from cyclophosphamide-immune suppressed mice (Endoxan, Baxter) at 6–7 days post-infection and mixed with defibrinated horse blood (E&O Laboratories Limited) to obtain > 10⁶ BSF trypanosomes/ml in the initial blood meal. After this trypanosome-infected blood meal, flies were fed every 2–3 days on uninfected defibrinated horse blood. Salivary gland infected flies were selected by induced probing on pre-warmed glass slides that were microscopically examined for the presence of metacyclic trypanosomes. Flies with a mature, metacyclic salivary gland infection (SG+) were selected for infecting mice. Some SG+ flies were dissected to isolate the salivary glands and to evaluate by flow cytometry the spontaneous parasitic outflow from non-disrupted glands in phosphate saline glucose buffer (PSG; PBS pH 7.4 supplemented with 1% glucose).