the final point of injection was previously confirmed by injection of 1 microliter of colorant in a small subgroup of animals. A stainless steel guide cannula, was inserted into the hole made previously. After penetrating the dura, we slowly lowered the cannula to the desired Z coordinate of the injection site, and 
once it reached the right depth slowly, 2 ml of solution were infused into the intracerebroventricular zone of the left brain hemisphere, using a single syringe infusion pump connected to the cannula via injection tubing previously filled with mineral oil. 25 minutes after the end of the infusion we retracted the cannula slowly to avoid backflow of the injected solution to the surface, and removed the animal from the stereotaxic frame. After cleaning the injection site with sterile saline by moist cotton swabs we sutured the skin with a non-absorbable, sterile, surgical silk suture and disinfected the scalp with Betadine along the incision site. Next, we injected sterile saline solution subcutaneously to avoid dehydration of the animal after the surgery, and subsequently we injected the same amount of glucosate solution to improve the animal feeding immediately after surgical procedure. Finally, we kept the animal warm on a temperature-controlled heating pad until its full recovery. Once the animal recovered, we returned it to a clean cage and put wet food pellets in the cage for easy access to food. Immunohistochemistry Under deep anesthesia, rats were perfused transcardially with a rinse of saline, PP-242 web followed by 4% formaldehyde fixative. Endogenous Hes3+ Cells in the Adult Hippocampus Brains were removed immediately, stored in the fixative solution overnight, and then in 30% sucrose for 3 days. Brains were frozensectioned at 12 or 30 micrometers. Immunohistochemical detection of BrdU was performed with an antigen-retrieval step. Wild type mice were deeply anesthetized and transcardially perfused with a saline solution, followed by 4% paraformaldehyde in Phosphate Buffer. Brains were removed, post-fixed in 4% paraformaldehyde in PB overnight and finally transferred in 30% sucrose in PB for 3 days. Brains were then coronally frozensectioned. Slices were rinsed three times at room temperature in PB, and then blocked in PB with 10% BSA, 0.3% Triton X-1000 for two hours. Sections were then incubated overnight at 4uC in PB with 0.3% Triton X-1000, 0.1% normal donkey serum with primary rabbit anti-Hes3 and mouse anti-Sox2 antibodies. Slices were then rinsed three times in PB at room temperature and incubated with Alexa Fluor 488-conjugated donkey anti-Mouse and DyLight 594-conjugated donkey anti-Rabbit secondary antibodies for 3.5 hrs at room temperature. Slices were rinsed three times in PB at room temperature and coverslipped in mounting medium. Immunofluorescence was then observed with a laser confocal microscope and images were acquired. ~~ A number of natural products, such as curcumin, isoflavone, resveratrol and epigallactocatechin-3-gallate, show efficacy in controlling the growth and metastasis of various cancers. Studies suggest that dietary intake of some of these products could aid in cancer prevention or enhance the efficacy of standard chemotherapeutic agents. Resveratrol is a polyphenolic antioxidant found in peanuts, grapes and red wine, which possesses significant health benefits. This compound has shown beneficial effects in experimental cancer models, where it suppresses the initiation, promotion and progression
Comments are closed.