Physical and mechanical properties of cardboard panels made

Physical and mechanical properties of cardboard panels made from used beverage carton with veneer overlay Ayrilmis, N. ; Candan, Z. ; Hiziroglu, S. 2008. 29, 1897– 1903 Student : Rattaphong Pokkaew (李平) Student ID : 0970456 Department of Food Science

Outline 1. Introduction 2. The objective 3. Methods 4. Results 5. Conclusion

Introduction

Peanut q The peanut (Arachis hypogaea L. ( q The peanut is called as the “king” of oil seeds. q Peanuts are also known as : • Ground nuts, earthnuts, goober peas, pindas, jack nuts, pinders, manila nuts and monkey balls. • In the UK these are sold as monkey nuts.

Tocopherols in peanut q Peanuts are a good source of tocopherols, phytosterol and phospholipid q The tocopherol content of peanuts varies with variety and production location q Peanut oil mainly contains -tocopherol 373– 50) ppm) and γ-tocopherols 390– 90)ppm( )Firestone, 1999(

Tocopherols in peanut q Sturm et al. (1966) determined tocopherol content of peanut oil from 17 varieties. Runner varieties had higher levels of -, γ-and -tocopherols than the Spanish varieties q Hashim et al. (1993) reported that there were significant differences in tocopherol content among Runner- and Virginia-type peanut cultivars

Phospholipid in peanut q Phospholipids (PL) contribute to the smoothness , texture, and mouthfeel of foods q Phospholipids improve the stability of the product because of their inherent antioxidant properties

Breast cancer q Breast cancer is the erratic growth of cells that originate in the breast tissue q A group of rapidly dividing cells may form a lump or mass of extra tissue. These masses are called tumors q The World Health Organization reported more than 1. 2 million people worldwide will be diagnosed with breast cancer this year

Breast cancer q The occurrence of breast cancer varies widely among women from different countries and cultures § Higher incidences in European and North American women § Lower in women in less developed countries and countries relying more heavily on vegetarian diets )American Cancer Society, 2003( q Dietary factors, specifically the proportion of animal versus plant fats, may play a role in the development of breast cancer ) Messina and Barnes, 1991; Cho et al. , 2003(

Breast cancer & Phytosterol q Plant foodstuffs contain specific phytochemicals which may offer protection from breast cancer. q An attractive hypothesis which may account for the cancer protective action of phytosterols is that phytosterols induce apoptosis or programmed cell death in highly proliferative tumor cells. q Plant foodstuffs contain specific phytochemicals which may offer protection from breast cancer.

Fig 1. Structure of some representative phytosterols Source : Moreau et al. , 2002; Berger et al. , 2004; Kritchevsky and Chen, 2005

Apoptosis q Apoptosis, or programmed cell death, is associated with several fundamental biological processes, including cell development, differentiation and response to injury. q Apoptosis is defined as a set of events that once initiated induce lethal changes that include membrane blebbing, mitochondrial break down and DNA fragmentation q Apoptosis occurs via two main pathways • The intrinsic or mitochondrial pathway • The extrinsic or death receptor-mediated pathway

Apoptotic proteaseactivating factor 1 (Apaf-1( Cytochrome c Fig 2. The intrinsic or mitochondrial pathway Pro-apoptotic factors

Fas ligand Fas receptor FADD Fas-associated death domain TNF receptorassociated death domain Fig 3. The extrinsic or death receptor-mediated pathway

The objective q The objective of this study was to assess the effect of cellular supplementation with either -sitosterol or cholesterol on the extrinsic caspase-8 pathway in the two breast cancer cell lines, MCF-7 and MDA-MB-231. q Hypothesized of -sitosterol, that may potentiate Fas-death domain signaling, leading to caspase-8 activation and ultimately to apoptosis. q To test the hypothesis, the breast cancer cell lines were treated with -sitosterol or with cholesterol as a control and effects on cell growth, sterol incorporation in cell membranes, expression of Fas-death domain signaling proteins, and caspase-8 activity were determined.

Methods

Cell culture MCF-7 and MDA-MB-231 cells 1% antibiotic–antimycotic 2 g/l sodium bicarbonate Cultured at 37 C 5% fetal bovine serum 5% CO 2 / 95% air as monolayers using RPMI 1640 growth medium

Preparation of sterol supplemented media and measurement of cell growth 2 -hydroxypropyl beta-cyclodextrin; CD complexes )RPMI 1640 media supplemented with cholesterol or b-sitosterol( Growth medium (8– 16 m. Msterols: 5 m. M CD) Control groups -Cell were treated with 5 m. M CD Studies groups

Measurement of sterol content of cell membranes by GLC Cells were seeded into 6 -well plates harvested by scraping frozen 80 C in 350 ml 10 m. M Tris and (20 m. M mannitol buffer (p. H 7. 4 Samples were thawed and briefly sonicated on ice

Samples were then saponified at 80 C in 95% ethanolic KOH 2 ml of hexane 2 ml of water Saponified samples The upper organic phase Dried under nitrogen The lower phase GLC

The media Determination of caspase-8 activity were replaced Cells were seeded in T-75 flasks; 10, 000)cells/cm 2( with sterolsupplemented 24 h or control media 3 day treatment Cells were scraped Washed in PBS (p. H 7. 4)

Lysed by suspension on ice of 710 cells/200 ml in buffer After 30 min The lysates were aspirated Centrifuged at 10, 000 g ; 4 C ; 30 min

The supernatants (cell lysates) were analyzed by the Bio-Rad DC protein assay Cell lysates (100 mg protein) were assayed caspase-8 activity

Results

Results Fig. 4. Effect of sterol supplementation on growth of MCF-7 cells. Cells were grown in RPMI-1640 growth medium supplemented with different concentrations of sterols

Results Fig. 5. Effect of sterol supplementation on growth of MDAMB-231 cells. Cells were grown in RPMI-1640 growth medium supplemented with different concentrations of sterols

Table 1. Effect of 2 d-sterol supplementation on sterol content of MCF-7 cell membranes* Supplementation ) Cholesterol mg/mg protein) -Sitosterol Total sterol -Sitosterol (mg/mg protein) (% total sterol( CD vehicle 43 2 a 0. 0 0 a 43 2 a 0 a Cholesterol 49 2 b 0. 0 0 a 49 2 a 0 a -Sitosterol 40 1 a 50 5 b 90 5 b 56 b cells were treated for 2 d with 16 m. M sterol or vehicle and sterol contents of membranes determined by gas–liquid chromatography as described. Data are means SEM (n = 3) and letters (a, b) of values in each column are significantly different (p<0. 05). *MCF-7

Table 2. Effect of 2 d-sterol supplementation on sterol content of MDA-MB-231 cell membranes* Supplementation ) Cholesterol mg/mg protein) -Sitosterol Total sterol -Sitosterol (mg/mg protein) (% total sterol( CD vehicle 49 7 a 0. 0 0 a 49 7 a Cholesterol 73 4 b 0. 0 0 a 73 4 b 0 -Sitosterol 33 2 a 51 6 b 84 8 b 0 a a 61 b cells were treated for 2 d with 16 m. M sterol or vehicle and sterol contents of membranes determined by gas–liquid chromatography as described. Data are means SEM (n = (3 and letters (a, b) of values in each column are significantly different (p<0. 05) *MDA-MB-213

Table 3. Effect of 3 d-sterol supplementation on caspase-8 activity of MCF-7 and MDA-MB-231 cell lysates* Supplementation Caspase-8 activity of MCF-7 lysates Caspase-8 activity of MDA-MB-231 lysates CD vehicle 6000 800 a 4600 200 a Cholesterol 5000 200 a 4600 200 a -Sitosterol 9800 400 b 8100 800 b were treated for 3 d with 16 m. M sterol or vehicle and caspase-8 activities of cell lysates determined. Data are RFU/100 mg lysate protein and represent means SEM (n = 3). Letters (a, b) of values in each column are significantly different (p<0. 05). *Cells

CONT CHOL SIT Fas. L FADD p-FADD Caspase-8 Actin Fig. 6. Effect of sterol supplementation on expression levels of Fas-related signaling proteins in MCF-7 cells were treated with sterols for 24 h as described. Expression of Fas-related signaling pathway proteins was quantified by immunoblot.

CONT CHOL SIT Fas. L FADD p-FADD Caspase-8 Actin Fig. 7. Effect of sterol supplementation on expression levels of Fas-related signaling proteins in MDA-MB-231 cells were treated with sterols for 24 h as described. Expression of Fas-related signaling proteins was quantified by immunoblot.

Conclusion

Conclusion q -sitosterol can be affect the amounts and activity of components of the extrinsic apoptotic pathway in human breast adenocarcinoma cells q -sitosterol induces a reduction in membrane sphingomyelin and an increase the ceramide levels in some tumor cells q The effect of -sitosterol treatment to increase caspase-8 activity and apoptosis in these cells may be mediated, at least in part, by changes in membrane sterol content and effects on the Fas apoptotic pathway

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Arachis hypogaea L Source : http: //upload. wikimedia. org/wikipedia/commons/1/1 f/Koeh-163. jpg http: //en. wikipedia. org/wiki/Image: Peanuts. jpg

Preparation of sterol supplemented media and measurement of cell growth Studies groups incubated for 24 h cells were seeded at 5000 cells/cm 2 into 24 -well plates media were replaced with that containing -sitosterol or cholesterol in graded concentrations or CD vehicle

Preparation of sterol supplemented media and measurement of cell growth Media were similarly changed on days 3 and 5 Cells were trypsinized and counted on days 2, 4 and 6 by Coulter Counter using the electrical sensing zone method Growth curves were generated from the Coulter Counter data