Specific Problems: 1. What stage in the maturity of the deadly nightshade fruit is most efficient as an anti-neoplastic agent? 2. What concentration of the extract is most efficient against HPV related cervical neoplasms? 3. What are the implications by the study should the crude polysaccharides derived from the product be proven a potent anti-neoplastic agent? HYPOTHESES: Is the Lubi-lubi extract a potent anti-neoplastic agent against HPV related cervical neoplasms? • Null hypothesis.
The lubi-lubi (Atropa belladonna) extract is not a potent anti-neoplastic agent against HPV related cervical neoplasms. • Operational hypothesis The lubi-lubi (Atropa belladonna) extract is a potent anti-neoplastic agent against HPV related cervical neoplasms. METHODOLOGY Materials: 1) For extracting the polysaccharide a) Lubi-lubi (Atropa belladonna) fruit in different levels of maturity b) distillation apparatusc) beakers d) petri dish e) refrigerator (for preservation of samples) f) mortar and pestle g) thermometer canoscopy.
2) for preperation and treatment of cancer cells a) sample of blood with HPV related neoplasms, 5ml per sample b) sample of normal blood, 5ml per sample c) centrifuge d) test tubes e) petri dish f) dropper g) refrigerator (for preservation of samples) h) compound microscope PROCEDURE 1. Collection of Lubi-lubi fruit - The plant to be used naturally grows in the province of Albay so collecting samples would not be hard. - To be collected are fruits in different levels of maturity: young, half ripe and ripe 200g each 2. Extracting fluid from the fruit a.
Using mortar and pestle, pound young fruits until fluid evidently separate from the fibers of the fruit b. Repeat for half ripe and ripe specimen c. Collect fluid and store in a beaker, label accordingly. 3. Dehydrating the fluid (rationale: to separate water from polysaccharide) a. Pour 5 ml of fluid (from young fruit) onto beaker, place thermometer in beaker. b. Heat beaker continuously at 100o c. Continue heating beaker until water on the distal end of the setup reaches 3. 75ml. d. Repeat process to get at the distal end of the setup 2. 5ml and 1.
25 ml respectively. Label accordingly. e. Repeat process for half ripe and ripe specimen. Label accordingly. 4. Testing the effect of isolated polysaccharide on neoplastic specimen a. Using the specimen collected by a professional, use Diffusion Kirby-Bauer2 disk-diffusion method to test effect of polysaccharide on neoplastic specimen. b. Swab specimen uniformly across a culture plate. c. A filter-paper disk, impregnated with the compound to be tested, is then placed on the surface of the agar. The compound diffuses from the filter paper into the agar.
The concentration of the compound will be highest next to the disk, and will decrease as distance from the disk increases. d. If the polysaccharide is effective against bacteria at a certain concentration, no further growth will be observed, or the spread of the neoplasm slows at a desirable rate. e. This is the zone of inhibition. Thus, the size of the zone of inhibition is a measure of the compound's effectiveness: the larger the clear area around the filter disk, the more effective the compound. EXPERIMENTAL METHOD The experimental method of research will be used in this study.
The experimental method is a systematic and scientific approach to research in which the researcher manipulates one or more variables, and controls and measures any change in other variables. This research is proposed to determine the efficacy of lubi-lubi fruit extract as an anti-neoplastic agent against HPV related cervical neoplasms. The advantages and disadvantages as well as the reliability of this instrument were also part of the objectives. Sample neoplastic cells are to be collected by biopsy from patients diagnosed with the condition. The biopsy would be conducted by a qualified oncologist.
The lubi-lubi (Atropa belladonna) extract is not a potent anti-neoplastic agent against HPV related cervical neoplasms. • Operational hypothesis The lubi-lubi (Atropa belladonna) extract is a potent anti-neoplastic agent against HPV related cervical neoplasms. METHODOLOGY Materials: 1) For extracting the polysaccharide a) Lubi-lubi (Atropa belladonna) fruit in different levels of maturity b) distillation apparatusc) beakers d) petri dish e) refrigerator (for preservation of samples) f) mortar and pestle g) thermometer canoscopy.
Using mortar and pestle, pound young fruits until fluid evidently separate from the fibers of the fruit b. Repeat for half ripe and ripe specimen c. Collect fluid and store in a beaker, label accordingly. 3. Dehydrating the fluid (rationale: to separate water from polysaccharide) a. Pour 5 ml of fluid (from young fruit) onto beaker, place thermometer in beaker. b. Heat beaker continuously at 100o c. Continue heating beaker until water on the distal end of the setup reaches 3. 75ml. d. Repeat process to get at the distal end of the setup 2. 5ml and 1.
25 ml respectively. Label accordingly. e. Repeat process for half ripe and ripe specimen. Label accordingly. 4. Testing the effect of isolated polysaccharide on neoplastic specimen a. Using the specimen collected by a professional, use Diffusion Kirby-Bauer2 disk-diffusion method to test effect of polysaccharide on neoplastic specimen. b. Swab specimen uniformly across a culture plate. c. A filter-paper disk, impregnated with the compound to be tested, is then placed on the surface of the agar. The compound diffuses from the filter paper into the agar.
The concentration of the compound will be highest next to the disk, and will decrease as distance from the disk increases. d. If the polysaccharide is effective against bacteria at a certain concentration, no further growth will be observed, or the spread of the neoplasm slows at a desirable rate. e. This is the zone of inhibition. Thus, the size of the zone of inhibition is a measure of the compound's effectiveness: the larger the clear area around the filter disk, the more effective the compound. EXPERIMENTAL METHOD The experimental method of research will be used in this study.
The experimental method is a systematic and scientific approach to research in which the researcher manipulates one or more variables, and controls and measures any change in other variables. This research is proposed to determine the efficacy of lubi-lubi fruit extract as an anti-neoplastic agent against HPV related cervical neoplasms. The advantages and disadvantages as well as the reliability of this instrument were also part of the objectives. Sample neoplastic cells are to be collected by biopsy from patients diagnosed with the condition. The biopsy would be conducted by a qualified oncologist.
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