An animal model of oral candidiasis was induced basically as reported by Jones et al., [15] (link) with some modifications. The rats were immunosuppressed, starting 1 week before experimental infection and continuing throughout the experiment, by administration of dexamethasone (Fortecortin; Merck Laboratories, Madrid, Spain) in the drinking water at a dose of 0.5 mg/l. Also, a 0.1% aqueous solution of tetracycline hydrochloride (Terramicine; Pfizer Laboratories, Alcobendas, Spain) was given to the rats, beginning 7 days before infection. The concentration of tetracycline hydrochloride were reduced to 0.01% on the day of infection and maintained throughout the experiment. A suspension of C.albicans, containing 5 × 108 viable cells/ml was prepared according to Reed et al. [16] (link). The suspension of C. albicans (0.2 ml) was dripped into the mouth of the rat with the aids of a 1 ml syringe and 30 × 8 mm blunt needle. Then the inoculum was spread on the tongue dorsum with a swab previously soaked in the suspension. The rats did not receive water for at least 1 h after inoculation. This procedure was repeated for 3 consecutive days. Establishment of C. albicans infection was evaluated by swabbing the inoculated oral cavity with sterile cotton applicator 6 days after inoculation, followed by plating on yeast extract- peptone- dextrose agar [4] (link).
Fortecortin
Manufactured by Merck Group
Sourced in Germany
Fortecortin is a lab equipment product manufactured by Merck Group. It is designed for use in scientific research and laboratory settings. The core function of Fortecortin is to provide a reliable and consistent tool for researchers to conduct their experiments and analyses.
Automatically generated - may contain errors
Lab products found in correlation
Glutamax, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Mem neaa 100, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Trypan blue exclusion technique, by Merck Group (1 mentions)
Neubauer chamber, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Six well culture plates, by BD (1 mentions)
Human insulin, by Sanofi (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Mem neaa 100x, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline solution, by Thermo Fisher Scientific (1 mentions)
Trypan blue, by Harvard Bioscience (1 mentions)
E coli 0127 b8, by Merck Group (1 mentions)
Diclofenac, by Merck Group (1 mentions)
8 protocols using fortecortin
1
Induction of Oral Candidiasis in Rats
2
Isolation of Primary Human Hepatocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue samples for PHH isolation were obtained from macroscopically tumor-free areas of resected livers (Table 1 ; D1–4 and DH2–10). PHHs were isolated as described previously [19 (link),20 (link),21 (link)] by a two-step EGTA/collagenase perfusion technique. Cells were pooled, washed with phosphate-buffered saline (PBS, Gibco, Paisley, UK) and resuspended in PHH culture medium (William’s Medium E, GlutaMAX™ (WME, Gibco, Paisley, UK) supplemented with 10% fetal bovine serum (FBS, Merck Biochrom, Berlin, Germany), 100 U/100 μM penicillin/streptomycin, 1% nonessential amino acids (MEM NEAA 100×), 15 mM HEPES, 1 mM sodium pyruvate (all provided by Gibco, Paisley, UK), 1 μg/mL dexamethasone (Fortecortin®, Merck, Darmstadt, Germany) and 80 IU/l human insulin (Lilly Deutschland GmbH, Bad Homburg, Germany/Sanofi-Aventis, Frankfurt am Main, Germany). Cell count and viability were determined using a Neubauer chamber with the trypan blue exclusion technique (Sigma-Aldrich, St. Louis, MO, USA).
3
2D Hepatocyte Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In parallel to bioreactor cultures, experiments with cells from the same donor were performed in 2-D plates for comparison. Six-well culture plates (Falcon; BD Biosciences, San Jose, CA, USA) were coated with rat-tail collagen prepared according to a protocol described previously.17 Freshly isolated human hepatocytes were suspended in Williams Medium E with GlutaMax (Thermo Fisher Scientific, Waltham, MA, USA), containing penicillin–streptomycin (100,000 U/L–100 mg/L), 15 mM HEPES, 1 mM sodium pyruvate, 1% non essential amino acids, 10% FBS (all Thermo Fisher Scientific), 0.8 mg/L dexamethasone (Fortecortin; Merck, Darmstadt, Germany), and 1 mM human insulin (Sanofi Aventis, Frankfurt am Main, Germany), referred to as “full medium”. Cells were seeded in the collagen-coated plates at a density of 1.4 million cells per well (Figure 3B and D ). From 24 hours onward, the culture medium was exchanged daily with the same medium, but without FBS and hormones (starving medium).
4
Establishing Rat Model for Candidiasis Study
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The study design was certified through approval from ethical board of the Faculty of Pharmacy, Damanhour University (Ref.No.720PM16). Female Sprague Dawley rats weighed 200–250 gm were utilized throughout the entire studies. They were given access to chow and water. They were habituated three per cage and maintained at 21 °C and 50–55% humidity under a 12:12 h light-dark cycle. Prior infection, all rats were subcutaneously injected with estradiol valerate dissolved in 0.10 µL sesame oil and given in a dose of 0.5 mg for each rat for six days to induce pseudo estrous (De Bernardis et al. 1997 (link)). Moreover, rats were administrated with Dexamethasone (Fortecortin; Merck Laboratories) (1 mg/L) as immunosuppressant and Amoxycillin (SAIMED innovation) (250 mg/L) in drinking water and continued through experiment (Martinez et al. 2001 (link)).
5
Isolation and Culture of Primary Human Hepatocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PHHs were isolated from liver tissue samples by a two-step EDTA/collagenase perfusion technique as described previously [47 (link),48 (link)]. The resulting PHH fractions were washed with PBS and re-suspended in PHH culture medium. Afterwards, the cell number and viability were determined in a Neubauer counting chamber by Trypan blue. The cells were seeded on cell culture plastics using hepatocyte culture medium for adherence overnight. PHH culture medium was based on William’s Medium E with GlutaMAX™ (WME, Gibco, Paisley, UK), supplemented with 10% fetal bovine serum (FBS, Merck Biochrom, Berlin, Germany), 15 mM HEPES, 0.1 mM non-essential amino acids (MEM NEAA 100×), 1 mM sodium pyruvate, 100 U/100 µM penicillin/streptomycin (all provided by Gibco, Paisley, UK), 80 IU/l human insulin (Lilly Deutschland GmbH, Bad Homburg, Germany), and 1 µg/mL dexamethasone (Fortecortin®, Merck, Darmstadt, Germany).
6
Neonatal Dexamethasone Exposure: Effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Male pups received s.c. injections of vehicle (saline) or DEX (Fortecortin, Merck, Darmstadt, Germany) on postnatal day (PND) 1–7 (DEX 200 μg kg−1 day−1 on PND 1–3, tapered down to 100 μg kg−1 day−1 on PND 4–7); the neonatal vehicle- and DEX-treated groups are hereafter referred to as CON and ND, respectively. Animals that showed signs of weakness or discomfort were immediately culled in accordance with rules on animal welfare.
7
Hepatocyte Attachment and Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The hepatocyte attachment and culture medium was based on William's Medium E with GlutaMAX™ (WME, Gibco, Paisley, UK), supplemented with 10 % fetal bovine serum (FBS, Merck Biochrom, Berlin, Germany), 15 mM HEPES, 0.1 mM non-essential amino acids (MEM NEAA 100x), 1 mM sodium pyruvate, 100 U/100 µM penicillin/streptomycin (all provided by Gibco, Paisley, UK), 80 IU/l human insulin (Lilly Deutschland GmbH, Bad Homburg, Germany) and 1 µg/ml dexamethasone (Fortecortin®, Merck, Darmstadt, Germany). Phosphate-buffered saline solution supplemented with calcium and magnesium (PBS) was purchased from Gibco and Trypan Blue was provided by Biochrom (Berlin, Germany). All other chemicals were purchased from Sigma-Aldrich (Munich, Germany), if not stated otherwise. Rat tail collagen was prepared in our laboratory according to the protocol established by Rajan et al. (2006[31 (link)]).
8
Endotoxin-Induced Inflammation Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Before the experiment was started, a stable baseline of MAP, ICP and LDF was secured. Arterial blood levels of O2, CO2, pH, K+ and Na+ were measured by the use of a blood analyzer (ABL 505; Radiometer, Copenhagen, Denmark) before and every 30 min during the experiment. Once a stable baseline was obtained LPS (E. Coli 0127 B8, Sigma-Aldrich) dissolved in sterile saline (1 mg/ml) or vehicle alone was injected i.p. (1 mg/kg) and subsequently i.v. (1 mg/kg). Ammonium acetate (NH3) at a concentration of 140 μmol/kg/min or saline alone was then administered i.v. to the animals by an infusion rate of 2.3–2.6 ml/hour and the chronometer was started.
Indomethacin (Confortid; Activas, Copenhagen, Denmark) or diclofenac (Sigma-Aldrich) 10 mg/kg were administered i.v. 15 min after start, while dexamethasone (Fortecortin; Merck, Darmstadt, Deutschland) 2 mg/kg was administered i.v. 30 min before start. Sterile saline was used as vehicle in the control group.
The experiment was terminated after 70 min. Arterial blood was sampled and the animal was decapitated. Blood samples were centrifuged and the plasma were frozen in liquid nitrogen in heparin containing tubes.
Indomethacin (Confortid; Activas, Copenhagen, Denmark) or diclofenac (Sigma-Aldrich) 10 mg/kg were administered i.v. 15 min after start, while dexamethasone (Fortecortin; Merck, Darmstadt, Deutschland) 2 mg/kg was administered i.v. 30 min before start. Sterile saline was used as vehicle in the control group.
The experiment was terminated after 70 min. Arterial blood was sampled and the animal was decapitated. Blood samples were centrifuged and the plasma were frozen in liquid nitrogen in heparin containing tubes.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!