As an integral part of the course, the Drug Interaction and Pharmacodynamics Laboratory provides students with practical experience in evaluating drug interactions at the pharmaceutical, pharmacokinetic and pharmacodynamic stages. Where [A] is the agonist concentration, nH is the Hill slope, and the EC50 value is the concentration of agonist that produces a response that is 50% of the maximal current. Bacteriostatics inhibit the growth of bacteria until the body's immune system takes over the rest of the bacteria.
One of the most popular methods for evaluating the synergistic, additive or antagonistic effects is the checkerboard and E-test method. The concentration of bacterial colonies was then matched to 0.5 McFarland turbidity standard (equivalent to bacterial concentration of 1.5x108 CFU/ml. Please check how to make McFarland standard at www.dalynn.com/dyn/ck_assets/files/tech/TM53. pdf!) by visual detection or by using spectrometer UV-Vis at 625 nm (absorbance. Before checkerboard testing, the bacterial colonies must be diluted with Mueller-Hinton broth medium until the concentration is 10x105 CFU/ml.
The MIC (minimal inhibitory concentration) will be interpreted as the lowest concentration of antibiotic that completely inhibits bacterial growth as detected by the naked eye. The MIC data will then be analyzed in terms of the fractional inhibitory concentration index (FIC). In which MIC A = minimal inhibitory concentration of A antibiotic; MIC B = minimal inhibitory concentration of B antibiotic; MICA combination A & B = minimal inhibitory concentration of combination A & B antibiotic, where the concentration of A is the smallest;.
MICB combination A and B = the minimum inhibitory concentration of the combination antibiotic A & B at which the concentration of B is the lowest.
Effect of Broccoli Juice Towards Ethanol-Induced Liver Damage
Transfer the tissue to xylene and paraffin (3x) for histology maintained at 60 ⁰C, respectively. Then immerse the slides in water before placing them in the hematoxylin and eosin jar for 2 minutes. Quickly rinse the slides with water and dehydrate the slides with increasing concentration of alcohol for 2 minutes.
Transfer the slides to xylene for 1 minute and cover the slides with an objective glass supplemented with eukitt.
Combination of Acetaminophen-Codeine as Analgesic in Mice
Twisting is defined as contraction of the abdominal muscle together with an extension of the hind legs or rotation of the trunk. please see https://www.youtube.com/watch?v=Ib63O4F856w for details). Example of solution calculation that will be given to mice. For example: the BW of the mouse is 30 mg. BW (g) Treatment given (name of drug, how many ml of drug, how many ml of gum . arabic solution).
Observe the number of writhes from acetaminophen-treated mice, codeine-phosphate-treated mice, and combination acetaminophen-codeine-phosphate-treated mice for a total of 5 minutes.
Pharmacokinetic Interaction Between Diazepam and Cimetidine
The amount of solution needed for each mouse is given orally to diazepam-treated group and diazepam-cimetidine-treated group, respectively. Note when the treatment is given and calculate the time when the mouse begins to sleep or loses its ability to grasp (drug onset) and the time when the mice wakes up or gains its ability to grasp (duration of drug effect). Record the time when the drug is given, when the mice start to sleep, and when the mice start to wake up by evaluating its grasping ability or turning the mice in a supine position.
Compare the duration of drug action between diazepam-treated mice and the combination of cimetidine-diazepam-treated mice. Discuss the interaction between cimetidine and diazepam related to the sedative, hypnotic/muscle relaxing effect of diazepam.
Combination of Taurine & Caffeine on Platelet Aggregation
Calculate the dose and solution required for each mouse using a dose of 13 mg/kg for caffeine and 130 mg/kg for taurine. The end of the mouse tail is cut off and the blood that comes out is absorbed into the capillary tube. Capillary containing blood is broken every 15 seconds until fibrin forms in the broken capillary.
Please explain about possible mechanisms by which caffeine, taurine and their combination can affect platelet aggregation.
Drug Incompatibility
Pharmacogenetic Polymorphisms of CYP2C19
Carefully apply 700 µL of the mixture from the previous step onto the QIAamp Mini spin column (in a 2 mL collection tube) without wetting the rim. Place the QIAamp Mini spin column in a clean 2 ml collection tube (supplied) and discard the filtrate tube. Carefully open the QIAamp Mini spin column and add 500 µL of buffer AW1 without wetting the rim.
Place the QIAamp Mini spin column in a clean 2 ml collection tube (provided) and discard the collection tube containing the filtrate. Carefully open the QIAamp Mini spin column and add 500 µL Buffer AW2 without wetting the rim. Place the QIAamp Mini spin column into a new 2 ml collection tube (not supplied) and discard the old collection tube with filters.
Place the QIAamp Mini spin column into a clean 1.5 ml microcentrifuge tube and discard the collection tube containing the filtrate. Carefully open the QIAamp Mini column and add 150 µL of buffer AE or distilled water. pure DNA is obtained at this stage). Genotyping of CYP2C19 is performed using the Polymerase Chain Reaction (PCR)-Restriction Fragment Length Polymorphism (RFLP) technique. Please see tutorial on YouTube videos about PCR and RFLP).
Dilute primers for CYP2C19*2 and CYP2C19*3 to 100 µM using Nuclease Free Water (1 primer set contains sense and antisense primers, respectively). Record the result from the nanodrop and take pictures of the electrophoresis gel, then interpret the data. Please explain the concept of Polymerase Chain Reaction (PCR)-Restriction Fragment Length Polymorphism (RFLP) technique and how it is used to detect genetic polymorphisms.
Describe the importance of cytochrome P450 enzymes, especially CYP2C19, and further explain the polymorphisms in CYP2C19, including its consequences.
Interaction of Cisapride and Erythromycin Induced Bradycardia in Danio rerio larval
The maximum amount of ethanol per well is 1% (1 well filled with 6 ml of medium. Maximum amount of ethanol is 60 µl per well). Harvest the embryo on the morning of day 1, collect live embryos and remove unfertilized and dead embryos. Dispense 2dpf zebrafish embryos into six-well microplates in 6mL fish water (supplemented with PTU 0.2 mM per well) containing the drug according to this treatment card.
In the normal state, the atrium and ventricle are side by side; blood flows into the atrium from the sinus venosus and out of the ventricle through the bulbus arteriosus. Morphological abnormalities, including the size of the heart and the number position of the heart chamber, should be investigated. Pericardial edema or pericardial effusion is an abnormal accumulation of fluid in the pericardial cavity.
Abnormalities in the heart will lead to abnormalities of blood circulation which can lead to other abnormalities including bleeding. Try to explain the action of each drug and the combination of the drugs on the heart of zebrafish larvae.
