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Extracellular RNAs and Extracellular Vesicles: Inception, Current Explorations, and Future Applications

  • Perumal, Ayyappasamy Sudalaiyadum;Chelliah, Ramachandran;Datta, Saptashwa;Krishna, Jayachandran;Samuel, Melvin S.;Ethiraj, Selvarajan;Park, Chae Rin
    • Journal of Food Hygiene and Safety
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    • v.35 no.6
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    • pp.535-543
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    • 2020
  • In addition to the ubiquitous roles of cellular RNA in genetic regulations, gene expression and phenotypic variations in response to environmental cues and chemotactic signals, the regulatory roles of a new type of RNA called extracellular RNAs (exRNAs) are an up-and-coming area of research interest. exRNA is transported outside the cell through membrane blebs known as membrane vesicles or extracellular vesicles (EVs). EV formation is predominant and conserved among all microbial forms, including prokaryotes, eukaryotes, and archaea. This review will focus on the three major topics concerning bacterially derived exRNAs, i.e., 1) the discovery of exRNA and influence of extraneous RNA over bacterial gene regulations, 2) the known secretion mechanism for the release of exRNA, and 3) the possible applications that can be devised with these exRNA secreted by different gram-negative and gram-positive bacteria. Further, this review will also provide an opinion on exRNA- and EV-derived applications such as the species-specific exRNA markers for diagnostics and the possible roles of exRNA in probiotics and the epigenetic regulations of the gut microbiome.

Inhibitory Effect of Biotransformed-Fucoidan on the Differentiation of Osteoclasts Induced by Receptor for Activation of Nuclear Factor-κB Ligand

  • Park, Bobae;Yu, Sun Nyoung;Kim, Sang-Hun;Lee, Junwon;Choi, Sung Jong;Chang, Jeong Hyun;Yang, Eun Ju;Kim, Kwang-Youn;Ahn, Soon-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.32 no.8
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    • pp.1017-1025
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    • 2022
  • Bone homeostasis is regulated by constant remodeling through osteogenesis by osteoblasts and osteolysis by osteoclasts and osteoporosis can be provoked when this balance is broken. Present pharmaceutical treatments for osteoporosis have harmful side effects and thus, our goal was to develop therapeutics from intrisincally safe natural products. Fucoidan is a polysaccharide extracted from many species of brown seaweed, with valuable pharmaceutical activities. To intensify the effect of fucoidan on bone homeostasis, we hydrolyzed fucoidan using AMG, Pectinex and Viscozyme. Of these, fucoidan biotransformed by Pectinex (Fu/Pec) powerfully inhibited the induction of tartrate-resistant acid phosphatase (TRAP) activity in osteoclasts differentiated from bone marrow macrophages (BMMs) by the receptor for activation of nuclear factor-κB ligand (RANKL). To investigate potential of lower molecular weight fucoidan it was separated into >300 kDa, 50-300 kDa, and <50 kDa Fu/Pec fractions by ultrafiltration system. The effects of these fractions on TRAP and alkaline phosphatase (ALP) activities were then examined in differentiated osteoclasts and MC3T3-E1 osteoblasts, respectively. Interestingly, 50-300 kDa Fu/Pec suppressed RANKL-induced osteoclasts differentiation from BMMs but did not synergistically enhance osteoblasts differentiation induced by osteogenic agents. In addition, this fraction inhibited the expressions of NFATc1, TRAP, OSCAR, and RANK, which are all key transcriptional factors involved in osteoclast differentiation, and those of Src, c-Fos and Mitf, as determined by RT-PCR. In conclusion, enzymatically low-molecularized 50-300 kDa Fu/Pec suppressed TRAP by downregulating RANKL-related signaling, contributing to the inhibition of osteoclasts differentiation, and represented a potential means of inducing bone remodeling in the background of osteoporosis.

The Experimental Study on Antioxidant, Anti-inflammatory, Antipruritic and Antibacterial Effects of the Banchong-san (BCS) (반총산의 항산화, 항염증, 항소양증, 항균효능에 관한 실험 연구)

  • Cho, Eun-Jin;Jo, Seong-Hui;Yang, Seung-Jeong
    • The Journal of Korean Obstetrics and Gynecology
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    • v.34 no.3
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    • pp.29-48
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    • 2021
  • Objectives: Banchong-san (BCS) is a herbal formula composed of 13 korean medicinal herbs and is traditionally used to treat inflammatory diseases and pain. The object of this study was to research the antioxidant, anti-inflammatory, antipruritic and antimicrobial effects of the BCS in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Methods: In this experiment, effects of BCS on the following four were measured as follows: (1) Anti-oxidative effects were evaluated by 1,1-diphenyl-2-picryl-hydrazyl (DPPH) Radical scavenging activity, 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) Radical scavenging activity. (2) Anti-inflammatory effects were evaluated by the production amount of Reactive oxygen species (ROS), Nitric oxide (NO), Interleukin-1β (IL-1β), Interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), Prostaglandin E2 (PGE2), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2)(the previous two are "mRNA"), extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinases (p38), inhibitor of nuclear factor kappa B (IκBα), nuclear factor kappa B (NF-κB) (the previous five are "Protein") in LPS-Stimulated RAW 264.7 cells. (3)Antipruritic effects were evaluated by the production amount of histamine, Leukotriene B4 (LTB4), LeukotrieneC4 (LTC4) Levels in phorbol 12-myristate 13-acetate(PMA)/ionomycin-stimulated MC/9 mast cell. (4) Anti-microbial effects were evaluated by the growth suppression of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Aspergillus niger. Results: The following results were obtained through each measurement: (1) DPPH Radical Scavenging Activity, ABTS Radical Scavenging Activity evoked a significant concentration-dependent increase. (2) ROS, NO, IL-1β, IL-6, TNF-α, PGE2 production amount, iNOS, COX-2 mRNA expression were significantly reduced in the BCS extraction group compared with the control group and significantly decreased the amount of ERK, JNK, p38, NF-κB Protein expression. The amount of IκB-α Protein Expression have increased significantly. (3) The amounts of histamine, LTB4, LTC4 were significantly decreased. (4) The antibacterial efficacy, BCS inhibited the growth of Escherichia coli, Pseudomonas aeruginosa at concentrations of 5 ㎍/ml, but did not suppress the growth of staphylococcus aureus and aspergillus niger. Conclusions: The experimental results show that BCS has anti-oxidant, anti-inflammatory, antipruritic and antimicrobial properties.

A Rapid Procedure for Screening and Isolation of Various Sizes of Plasmid DNA in Serovars of Bacillus thuringiensis (Bacillus turingiensis 변종(變種)들로부터의 Plasmid DNA 추출(抽出) 및 분리(分離))

  • LEE, YUNG KEUN;Faust, Robert M.;KANG, SEOK KWON;McCawley, Patricia E.;Meyers-Dowling, Carol L.
    • Korean journal of applied entomology
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    • v.24 no.1 s.62
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    • pp.45-50
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    • 1985
  • The use of a modified procedure for the isolation of extrachromosomal DNA of low to high molecular weight, followed by agarose gel electrophoresis of the crude lysates, provided a simple screening procedure for detecting plasmids ranging in molecular weights from approximately 1 to more than 135 megadaltons from serovars of Bacillus thuringiensis. The procedure provides for a relatively large-volume stable lysate for isolation of plasmids for restriction endonuclease mapping and cloning procedures. The method was used for screening of plasm ids in 6 differenentially effective serovars of B. thuringiensis toxic to dipteran and lepidopteran insects. Relatively large plasmid DNAs of masses above 50 megadaltons (Mdal) were isolated from all of the serovars examined using this technique. The number of extrachromosomal DNAs detected in serovars of B. thuringiensis was 8 for israelensis, 10 for kurstaki, 13 for aizawai, 2 for dendrolimus, 1 for finitimus, and 6 for yunnanensis. Smaller plasmid DNAs were isolated in four of the six serovars that ranged in mass down to approximately 2 Mdal.

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Major, Trace and Rare Earth Element Geochemistry, and Oxygen-Isotope Systematics of Illite/smectite in the Reindeer D-27 Well, Beaufort-Mackenzie Basin, Arctic Canada (카나다 보포트-맥켄지 분지의 일라이트/스멕타이트의 원소 지화학 및 산소동위원소 연구)

  • Ko, J.;Hesse, R.;Longstaffe, F.J.
    • Economic and Environmental Geology
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    • v.28 no.4
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    • pp.351-367
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    • 1995
  • The elemental geochemistry and oxygen isotopes of illite/smectite (I/S) have been studied in relationship to the mineralogical trend in the Reindeer D-27 well, Beaufort-Mackenzie Basin. The increase in concentrations of $K_2O$, Rb and rare earth elements (REE), the decrease in concentrations of tetrahedral elements such as Mg, Ti, Sc, Zn and Zr, and the increase in concentrations of tetrahedral elements such as Be and V can be related to I/S compositions that vary systematically with depth. Layer formulae of S- and I-layers are estimated as $[Al_{1.57}Fe_{.19}Mg_{.31}Ti_{.07}][Si_{3.84}Al_{.16}]O_{10}(OH)_2$ and $[Al_{1.84}Mg_{.16}][Si_{3.33}Al_{.67}]O_{10}(OH)_2$, respectively. The mobilization of REE appears to occur during illitization. The increase in concentrations of REE, especially La and Ce, with depth is probably linked to incorporation of ions with high valency (e.g. $V^{5+}$) in tetrahedral sites. The excess valency due to V is partly counter-balanced by ions with low valency (e.g. $Be^{2+}$) and, in turn, the local valency deficiency caused by $Be^{2+}$ could be compensated by high-charge interlayer cations such as REE (+3). ${\delta}^{18}O$ values of I/S range from 2.91 to 15.72‰ (SMOW), and increase with depth, contrasting to trends observed in the Gulf Coast and elsewhere. The increase in ${\delta}^{18}O$ of I/S results from the rapid increase in ${\delta}^{18}O$ of pore water that overcomes the decrease in temperature-dependent fractionation values with increasing burial depth (${\delta}^{18}O_{pore\;water}>-d{\Delta}/_{I/S-water};\;d{\delta}^{18}O_{I/S}>0$). Calculated ${\delta}^{18}O$ values of pore water in equilibrium with I/S suggest that the original water was probably meteoric water. The stratification of pore water is postulated from the presence of an isotopically light interval, about 450m thick. The depth range of the isotopically light zone overlaps, but does not coincide with the interval of lowered I-content and $K_2O$ concentrations, suggesting that oxygens may have been exchanged independently of mineralogical and geochemical reactions.

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Industrial potential of domestic Zanthoxylum piperitum and Zanthoxylum schinifolium: Protective effect of both extracts on high glucose-induced neurotoxicity (국내산 초피와 산초의 산업적 활용 가능성: 고당으로 유도된 뇌신경세포 독성에 대한 추출물의 보호 효과)

  • Han, Hye Ju;Park, Seon Kyeong;Kim, Min Ji;An, Jun Woo;Lee, Se Jin;Kang, Jin Yong;Kim, Jong Min;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.52 no.3
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    • pp.274-283
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    • 2020
  • This study focused on the in vitro investigation of antioxidant and anti-diabetic activities, along with neuroprotection against high glucose-induced cytotoxicity, in order to evaluate the physiological effects of Zanthoxylum piperitum and Zanthoxylum schinifolium. The highest total phenolic content was measured in the 40% ethanolic extracts of Zanthoxylum piperitum (EZP) and Zanthoxylum schinifolium (EZS). The in vitro EZP antioxidant activity showed a relatively higher ABTS/DPPH radical scavenging activity and malondialdehyde inhibitory effect than that of EZS. The EZP inhibited carbohydrate hydrolysis (α-glucosidase and α-amylase) more efficiently than EZS in anti-diabetic tests. However, EZS showed a more efficient inhibition of advanced glycation end-products formation than EZP. In addition, both EZP and EZS effectively protected human-derived neuronal cells from high glucose-induced cytotoxicity. Finally, the physiological compounds were analyzed using UPLC IMS-QTOF/MSE, and the main EZP (quercetin-3-O-glucoside and 3-caffeoylquinic acid) and EZS (5-caffeoylquinic acid) compounds were identified as phenolic compounds.

Study for the Synthesis of $[^{123}I]$Idoxifene and Its Uptake in the Breast Cancer Cell ($[^{123}I]$Idoxifene 합성과 유방암의 세포섭취에 관한 연구)

  • Cho, Young-Sub;Yang, Seung-Dae;Suh, Yong-Sup;Chun, Kwon-Soo;Ahn, Soon-Hyuk;Lim, Soo-Jung;Lim, Sang-Moo;Yu, Kook-Hyun
    • The Korean Journal of Nuclear Medicine
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    • v.34 no.5
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    • pp.410-417
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    • 2000
  • Purpose: Idoxifene is currently entering phase II clinical trials for the treatment of advanced breast cancer. The radiolabeled idoxifene using $[^{123}I]$ provides an opportunity for clinical pharmacology with single photon emission computed tomography (SPECT). The purpose of this study was to prepare radiolabeled idoxifene using $[^{123}I]$ and to determine its cell uptake of breast cancer cell line. Materials and Methods: With a view to evaluating new anticancer drugs, we are investigating the novel antiestrogen pyrrolidino-4-iodotamoxifen (idoxifene). $[^{123}I]$Idoxifene has been prepared in no-carrier-added form using a tributyl stannylated precursor which has been synthesized by means of (2-chloroethoxy)benzene with (${\pm}$)-2-phenylbutanoic acid on the basis of previously reported standard methods. The biodistribution and dynamic behavior of the compound were investigated using the comparative breast cancer cell line, MCF-7 (estrogen receptor-positive) and MDA-MB-468 (non-estrogen receptor). Results and Conclusion: Acylation of (2-chloroethoxy)benzene with (${\pm}$)-2-phenylbutanoic acid gave the versatile ketone (81%) which reacted with 1,4-diiodobenzene to give triphenylethylene as a mixture of E and Z geometric isomers, which were separated by the recrystallization in ethanol. The E-isomer was treated with pyrrolidine to give idoxifene (67%). In order to incorporate radioactive iodine into the 4-position, the 4-stannylated precursor was prepared (30%). The yield of radioiodination was 90-92% with a high radiochemical purity greater than 98%. The ratio of tumor uptake of the breast cancer cell line between MCF-7 and MDA-MB-468 was about 1.7.

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Potential Contamination Sources on Fresh Produce Associated with Food Safety

  • Choi, Jungmin;Lee, Sang In;Rackerby, Bryna;Moppert, Ian;McGorrin, Robert;Ha, Sang-Do;Park, Si Hong
    • Journal of Food Hygiene and Safety
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    • v.34 no.1
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    • pp.1-12
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    • 2019
  • The health benefits associated with consumption of fresh produce have been clearly demonstrated and encouraged by international nutrition and health authorities. However, since fresh produce is usually minimally processed, increased consumption of fresh fruits and vegetables has also led to a simultaneous escalation of foodborne illness cases. According to the report by the World Health Organization (WHO), 1 in 10 people suffer from foodborne diseases and 420,000 die every year globally. In comparison to other processed foods, fresh produce can be easily contaminated by various routes at different points in the supply chain from farm to fork. This review is focused on the identification and characterization of possible sources of foodborne illnesses from chemical, biological, and physical hazards and the applicable methodologies to detect potential contaminants. Agro-chemicals (pesticides, fungicides and herbicides), natural toxins (mycotoxins and plant toxins), and heavy metals (mercury and cadmium) are the main sources of chemical hazards, which can be detected by several methods including chromatography and nano-techniques based on nanostructured materials such as noble metal nanoparticles (NMPs), quantum dots (QDs) and magnetic nanoparticles or nanotube. However, the diversity of chemical structures complicates the establishment of one standard method to differentiate the variety of chemical compounds. In addition, fresh fruits and vegetables contain high nutrient contents and moisture, which promote the growth of unwanted microorganisms including bacterial pathogens (Salmonella, E. coli O157: H7, Shigella, Listeria monocytogenes, and Bacillus cereus) and non-bacterial pathogens (norovirus and parasites). In order to detect specific pathogens in fresh produce, methods based on molecular biology such as PCR and immunology are commonly used. Finally, physical hazards including contamination by glass, metal, and gravel in food can cause serious injuries to customers. In order to decrease physical hazards, vision systems such as X-ray inspection have been adopted to detect physical contaminants in food, while exceptional handling skills by food production employees are required to prevent additional contamination.

Culture Conditions of Aspergillus oryzae in Dried Food-Waste and the Effects of Feeding the AO Ferments on Nutrients Availability in Chickens (건조한 남은 음식물을 이용한 Aspergillus oryzae균주 배양조건과 그 배양물 급여가 닭의 영양소 이용률에 미치는 영향)

  • Hwangbo J.;Hong E. C.;Lee B. S.;Bae H. D.;Kim W.;Nho W. G.;Kim J. H.;Kim I. H.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.291-300
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    • 2005
  • Two experiments were carried out to assess the appropriate incubation conditions namely; duration, moisture content and the ideal microbial inoculant for fermented dried food waste(EW) offered to broilers. The nutrient utilization of birds fed the FW diets at varying dietary inclusion rates was also compared with a control diet. In Experiment 1, different moisture contents(MC) of 30, 40, 50 and $60\%$ respectively were predetermined to establish the ideal duration of incubation and the microbial inoculant. A 1mL Aspergillus oryzae(AO) $(1.33\times10^5\;CFU/mL)$ was used as the seed inoculant in FW. This results indicated that the ideal MC for incubation was $40\~50\%$ while the normal incubation time was > 72 hours. Consequently, AO seeds at 0.25, 0.50, 0.75 and 1.00mL were inoculated in FW to determine its effect on AO count. The comparative AO count of FW incubated for 12 and 96 hours, respectively showed no significant differences among varying inoculant dosage rates. The FW inoculated with lower AO seeds at 0.10, 0.05 and 0.01mL were likewise incubated for 72 and 96 hours, respectively and no changes in AO count was detected(p<0.05). The above findings indicated that the incubation requirements for FW should be $%40\~50\%$ for 72 hours with an AO seed incoulant dosage rate of 0.10mL. Consequently, in Experiment II, after determining the appropriate processing condition for the FW, 20 five-week old male Hubbard strain were used in a digestibility experiment. The birds were divided into 4 groups with 5 pens(1 bird per pen). The dietary treatments were; Treatment 1 : Control(Basal diet), Treatment 2 : $60\%$ Basal+4$40\%$ FW, Treatment 3 : $60\%$ $Basal+20\%\;FW+20\%$ AFW(Aspergillus oryzae inoculate dried food-waste diet) and Treatment 4: $60\%$ Basal+$40\%$ Am. Digestibility of treatment 2 was lowed on common nutrients and amino acids compared with control(p<0.05) and on crude fat and phosphorus compared with AFW treatments(T3, T4)(plt;0.05). Digestibility of treatment 3 and 4 increased on crude fiber and crude ash compared treatment 2 (p<0.05). Digestibility of control was high on agrinine, leucine, and phenylalnine of essential amino acids compared with treatment 3 and 4(p<0.05), and diestibility of treatment 3 and 4 was improved on arginine, lysine, and threonine of essential amino acids. Finally, despite comparable nutrient utilization among treatments, birds fed the dietary treatment containing AO tended to superior nutrient digestion to those fed the $60\%$ Basa1+$40\%$ FW.

Consumer Awareness and Evaluation of Retailers' Social Responsibility: An Exploratory Approach into Ethical Purchase Behavior from a U.S Perspective (소비자인지도화령수상사회책임(消费者认知度和零售商社会责任): 종미국시각출발적도덕구매행위적탐색성연구(从美国视角出发的道德购买行为的探索性研究))

  • Lee, Min-Young;Jackson, Vanessa P.
    • Journal of Global Scholars of Marketing Science
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    • v.20 no.1
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    • pp.49-58
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    • 2010
  • Corporate social responsibility has become a very important issue for researchers (Greenfield, 2004; Maignan & Ralston, 2002; McWilliams et al., 2006; Pearce & Doh 2005), and many consider it necessary for businesses to define their role in society and apply social and ethical standards to their businesses (Lichtenstein et al., 2004). As a result, a significant number of retailers have adopted CSR as a strategic tool to promote their businesses. To this end, this study sought to discover U.S. consumers' attitudes and behavior in ethical purchasing and consumption based on their subjective perception and evaluation of a retailer. The objectives of this study include: 1) determine the participants awareness of retailers corporate social responsibility; 2) assess how participants evaluate retailers corporate social responsibility; 3) examine whether participants evaluation process of retailers CSR influence their attitude toward the retailer; and 4) assess if participants attitude toward the retailers CSR influence their purchase behavior. This study does not focus on actual retailers' CSR performance because a consumer's decision making process is based on an individual assessment not an actual fact. This study examines US college students' awareness and evaluations of retailers' corporate social responsibility (CSR). Fifty six college students at a major Southeastern university participated in the study. The age of the participants ranged from 18 to 26 years old. Content analysis was conducted with open coding and focused coding. Over 100 single-spaced pages of written responses were collected and analyzed. Two steps of coding (i.e., open coding and focused coding) were conducted (Esterberg, 2002). Coding results and analytic memos were used to understand participants' awareness of CSR and their ethical purchasing behavior supported through the selection and inclusion of direct quotes that were extracted from the written responses. Names used here are pseudonyms to protect confidentiality of participants. Participants were asked to write about retailers, their aware-ness of CSR issues, and to evaluate a retailer's CSR performance. A majority (n = 28) of respondents indicated their awareness of CSR but have not felt the need to act on this issue. Few (n=8) indicated that they are aware of this issue but not greatly concerned. Findings suggest that when college students evaluate retailers' CSR performance, they use three dimensions of CSR: employee support, community support, and environmental support. Employee treatment and support were found as an important criterion in evaluation of retailers' CSR. Respondents indicated that their good experience with a retailer as an employee made them have a positive perception and attitude toward the retailer. Regarding employee support four themes emerged: employee rewards and incentives based on performance, working environment, employee education and training program, and employee and family discounts. Well organized rewards and incentives were mentioned as an important attribute. The factors related to the working environment included: how well retailers follow the rules related to working hours, lunch time and breaks was also one of the most mentioned attributes. Regarding community support, three themes emerged: contributing a percentage of sales to the local community, financial contribution to charity organizations, and events for community support. Regarding environments, two themes emerged: recycling and selling organic or green products. It was mentioned in the responses that retailers are trying to do what they can to be environmentally friendly. One respondent mentioned that the company is creating stores that have an environmentally friendly design. Information about what the company does to help the environment can easily be found on the company’s website as well. Respondents have also noticed that the stores are starting to offer products that are organic and environmentally friendly. A retailer was also mentioned by a respondent in this category in reference to how the company uses eco-friendly cups and how they are helping to rebuild homes in New Orleans. The respondents noticed that a retailer offers reusable bags for their consumers to purchase. One respondent stated that a retailer uses its products to help the environment, through offering organic cotton. After thorough analysis of responses, we found that a participant's evaluation of a retailers' CSR influenced their attitudes towards retailers. However, there was a significant gap between attitudes and purchasing behavior. Although the participants had positive attitudes toward retailers CSR, the lack of funds and time influenced their purchase behavior. Overall, half (n=28) of the respondents mentioned that CSR performance affects their purchasing decisions making when shopping. Findings from this study provide support for retailers to consider their corporate social responsibility when developing their image with the consumer. This study implied that consumers evaluate retailers based on employee, community and environmental support. The evaluation, attitude and purchase behavior of consumers seem to be intertwined. That is, evaluation is based on the knowledge the consumer has of the retailers CSR. That knowledge may influence their attitude toward the retailer and thus influence their purchase behavior. Participants also indicated that having CSR makes them think highly of the retailer, but it does not influence their purchase behavior. Price and convenience seem to surpass the importance of CSR among the participants. Implications, recommendations for future research, and limitations of the study are also discussed.