EVALUATION OF ANALGESIC AGENTS: THERMAL STIMULUS Hot-plate test Mice on hot plate at 55°c. cut off time 30 sec PHYSICAL STIMULUS Haffner’s tail clip method A metal artery clip was applied to the root of the mouse’s tail for 30 sec.
Writhing test the mice were given an intraperitoneal (i.p.) injection of 0.6% v/v acetic acid in a volume of 10ml/kg to induce the characteristic writhings.
Carrageenan-Induced Rat Paw Edema:Animals of group I received normal saline (3 ml/kg b.w. intraperitoneal, i.p) and served as saline control.The .groups II and III received methanol extract of . Clerodendron infortunatum (250 and 500 mg/kg b.w., i.p, . respectively) and group IV received reference drug . phenylbutazone (100 mg/kg b. w., i.p)..
EHRLICH ASCITES CARCINOMA(EAC) weekly intraperitoneal inoculation of 106 cells per mouse. Substances under investigation are administered either orally or intraperitoneally. The viability of the cells are judged by the trypan blue test viable tumour cell count is done using haemocytometer.
is induced in Balb-C mice by subcutaneous injection of 3-methylcholanthrene(3-MC).
defn The capability of producing fetal malformation. A given teratogen may be organ specific. It may be species specific. It can be dose specific. teratogens halidomide Accutane Diethylstilbestrol(DES) Alcohol OBSERVATION PARAMETERS
Multigenerational studies Single generational studies Segment I:Evaluation of Fertility and Reproductive Performance Segment II: Assessment of Developmental Toxicity Segment II: Postnatal Evaluation Tests under ICH: Fertility Assessment Postnatal Evaluation and Pregnancy State Susceptibility Assessment of Developmental Toxicity
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Mutation A change in the DNA molecule Process which produces changes in the DNA that may be inherited. MECHANISM 1. By the change in the structure of DNA which is responsible in inaccurate application of genome. 2. Prevent cell proliferation that fixes the DNA damage. 3. DNA repair removal of large segement of DNA.
Induction of in vitro mammalian cells resistant to certain chemical compound ◦ (e.g. 8-azaguanine, 6-thioguanin, ouabain) SOS/umu test – measurement of DNA repair in bacteria ◦ The bacteria is transformed by plasmide with mutator (repair) genes umuC and umuD fused with the gene for galactosidase. If the DNA is damaged by mutagen both umu mutator genes and gal gene are transcribed and galactosidase turns a specific substrate into a color product.
Test system – auxotrophic strain of Salmonella typhimurium – survives only in medium with histidine (dies in normal medium without histidine) After treatment with mutagen some auxotrophic cells are turned into normal ones that synthesize histidine and survive in a normal medium. These cells are called revertants (due to reverse mutation).
Mutagen His– Reverse mutation His+ bacteria bacteria Medium Dies in a Normal containing normal medium histidine medium
0 Negative control SPONTANEOUS REVERTANTS A dish with GENOTOXICITY a CONFIRMED compound to be tested Positive IS USED FOR CONTROL OF control THE TEST
Number of revertant colonies Dose of 1-nitropyrene (micrograms per dish) The graph describes results of the Ames test of various 1-nitropyrene doses. Describe the relationship between the dose and number of revertant colonies. Is it possible to confirm the mutagenic activity? Why we test the compound in various concentrations?