The sample was transferred to a 250 ml conical flask kept in water bath for alkali treatment. 75 ml of 17.5% caustic soda was measured using a measuring cylinder at 20°C. 15 ml of 17.5% NaOH was added and fibres were macerated gently with a flattened glass rod for 1 minute. 10 ml more NaOH was added and the solution was mixed for 45 seconds. 10 ml NaOH was again added and mixed for 15 seconds to make lump free slurry.
The chemical compound and the water were mixed together using a glass rod until the chemical was thoroughly dissolved. After the chemical had dissolved, the beaker containing 20mL of room temperature water was placed into the chemical mixture with a thermometer in the beaker as well. The lid for the polystyrene cup was then placed onto the cup, and the stopwatch began timing. The lid was not removed until exactly 2 minutes had passed, and the thermometer was taken out. The final temperature was recorded and the difference in temperature change was calculated by subtracting the initial water temperature from the final water temperature.
22.5 g of plate count agar powder was dissolved in a litre of sterile distilled water on the hot plate 2. pH of the solution was adjusted to 7.0 ± 0.2 by adding NaOH or HCl and was immediately transferred into the Schott bottle to be autoclaved at 121 ° C for 15 minutes 3. Prepared medium was stored in 4° C chiller Lauryl Sulphate Broth
Water will act as initial solvent for caffeine extraction. This is due to water that slowly soluble with caffeine at ambient temperature but highly soluble when temperature is at 100°C. Then, methylene chloride is chosen as the extraction solvent, due to its miscibility with caffeine and immiscibility with water. As mentioned above, the immiscible pair is chose for the extraction part because to allow the aqueous and organic layers to be separated. Basically, the bottom layer is the aqueous layer while the upper layer is the organic compound.
Twenty tablets were weighed accurately and powdered. An amount of the powder equivalent to 5 mg of amoxicillin trihydrate (content of one tablet) was dissolved in 60 ml of diluent. The solution was stirred for 10 min using a magnetic stirrer and filtered into a 100 ml volumetric flask through 0.45µ nylon membrane filter. The residue was washed 3 times with 10 ml of diluent and then the volume was completed to 100 ml with the same solvent. This solution was diluted with diluents to gae a concentration of 0.1 mg/ml solution each of Amoxicillin trihydrate.
Measurement of lipid peroxidation TBARS, a measure of lipid per oxidation, was measured as described by Ohkawa . Briefly, 1 ml of suspension medium was taken from the 10% tissue homogenate. 0.5 ml of 30% Trichloroacetic acid (TCA) was added to it, followed by 0.5 ml of 0.8% thiobarbituric acid (TBA) reagent. The tubes were covered with aluminium foil and kept in shaking water bath for 30 minutes at 80°C. After 30 minutes, tubes were taken out and kept in ice-cold water for 30 minutes.
Standardization of NaOH solution Accurately weigh out a sample of approximately 0.3-0.4 g of primary standard potassium hydrogen phthalate, KHPh, which has been previously dried at 120°C. Do not use more than 0.4 g. To obtain an accurate mass, weigh the sample on weighing paper, slide it into a clean (but not necessarily dry) 250 mL Erlenmeyer flask and reweigh the paper to account for any KHPh that may remain on it. Dissolve the KHPh sample in about 50 mL of CO2-free water and add 2-3 drops of 0.1% phenolphthalein indicator. Begin adding the approximately 0.1 M sodium hydroxide solution from the buret while continuously swirling the flask contents. Do not open the stopcock completely.
MATERIALS AND METHODS Bacterial strains and culture conditions Two S. aureus strains were used in the present study; S. aureus 8325-4 (SigB-) and SH1000 representing a SigB+.strain. Overnight cultures were grown in Luria Broth (LB) at 37°Cwith shaking at 150 rpm. Exposure to antibiotics was carried out as detailed below. Antibiotics Ciprofloxacin were purchased from Sigma-Aldrich CO. 10 mg/ml stock solution of antibiotic were prepared freshly with 0.1N HCl and stored at -20°C. During the experiment we diluted with sterile water 1:10 and 1:100 depending on the different drug concentration.
Fe3O4 nanoparticles were synthesized according to our previously reported method by chemical co-precipitation of Fe2+ and Fe3+ ions with a molar ratio of 1:2 . Briefly, 2.4 g of FeCl3.6H2O and 0.8 g of FeCl2.4H2O were dissolved in 30 mL of deionized water under using continuous N2 purge at 70 °C. and Under vigorous stirring, followed by dropwise addition of 10 mL of NH3.H2O was dropwise added to the reaction mixture until the color of mixture turned to black and kept reacting for 90 min to complete the reaction. At the end, the synthesized Fe3O4 nanoparticles were separated by a magnet and washed by using water and ethanol for further three times with 89.3%
Preparation of hot water extracts: 20 g from each bark powder was surface sterilized and mixed with 100 ml sterilized water and placed in water path for 90 minutes at 100ºC, and then filtered through 3 layers of sterilized cheese cloth. The products were kept in a refrigerator at 4 ºC until needed for testing their bacterial
Place C Elegans into the plates with the E Coli and leave for 24 hours. Examine the C Elegans to insure that the C Elegans have survived at the room temperature and continue to have multiple C Elegans surviving. Once this is done prepare the dilutions of all the subjects which we are testing. Start with 1% solution for Nitrate-N 100ml and move 10ml of the first well into the next. Fill the well with 90ml dh20 to reach 100ml.
The dried roots of Inula racemosa were pulverized and sieved with 100 ~ 200 mesh. The herb powder was placed into a glass bottle. Ultrasound-assisted extraction was carried out in an ultrasonic cleaner RK102H (Bandelin sonorex, Germany). The powder of Inula racemosa was extracted three times under the following conditions: the ratio of material to solvent was 10:1, undergoing ultrasonic treatment 30 minutes at 25 °C, 100 kHz /450 W.31 Before large extraction, a small-scale extraction experiments were carried out: 95% ethanol and ethyl acetate as the extractive solutions was investigated, respectively. The extraction efficiency was evaluated according to the percent content of AL and IS contained in the dried roots of Inula racemosa and calculated