Identification of Epitope in EAV N Protein

Identification of Epitope in EAV N ProteinIdentification of a novel maintain B cell epitope in the N protein of EAV (Bucyrus strain)Running title Identification of epitope in EAV N protein.HighlightsOne EAV N-specific mAb 1C11 was positive.A marginal linear peptide epitope within the N protein was determine.The determine epitope was conserved among several(predicate) regional EAV strains.The mAb and identified epitope may be recyclable diagnostic tools for EAV infection.AbstractObjective To identify the minimal epitope of N protein of the equid arteritis virus (EAV).Methods The full-length sequence of EAV N gene was cloned by RT-PCR and ligated into pET32a vector for expression. The recombinant pET-N protein was uttered in E. coli and purified by Ni affinity chromatography. The purified N protein was utilise to immunize mice for preparing monoclonal antibody (mAb). The responsiveness of mAb was evaluated by westerly blot and immunofluorescence assay (IFA). The peptides were identified utilize the fain mAb by indirect ELISA and Western blot. The homology analysis was performed using DNAMAN software.Results Recombinant EAV N protein was successfully expressed in the procaryon expression system. An EAV N-reactive mAb was selected and designated as 1C11. Indirect ELISA results showed that overlapping domain of MBP-N10 and MBP-N11 was recognized by the mAb 1C11. Further, the indirect ELISA and Western blot showed that 101QRKVAP106 was the minimal linear epitope of the EAV N protein. The homology analysis showed that the identified epitope is conserved among all EAV isolated strains, with the exception of the ARVAC which is a modified live virus vaccinum strain.Conclusion One EAV N-specific mAb was developed and a minimal linear peptide epitope within the N protein was identified. The EAV N-specific mAb and the defined linear peptide epitope of EAV N protein may be useful for the ontogenesis of specific diagnostic tools and design of vaccine.Keywords Epi tope N protein Equine arteritis virus Monoclonal antibodyIntroductionEquine arteritis virus (EAV) is the etiologic agent of equine viral arteritis (EVA) which is a respiratory and reproductive disease of horses 1-3. EAV was rst isolated from horses in Ohio in 1953. It is the prototype virus of the family Arteriviridae (genus Arterivirus, order Nidovirales) 4, 5. EAV infection of horses has been reported in many countries including New Zealand, Australia, and southwestward Africa 6-10.EAV is a positive-sense, enveloped and single-stranded RNA molecule with a length of 12.7kb 11. It contains devil large open reading frames (ORFs, 1a and 1b) and seven smaller ORFs (2a, 2b, and 3 to 7). ORFs 1a and 1b encode two replicase polyproteins (pp1a and pp1ab), whereas the ORFs 2a, 2b, 5, 6, and 7 encode the known EAV structural proteins E, GS, GL, M, and N, respectively 12. Moreover, ORFs 3 and 4 encode glycosylated membrane-associated proteins whose functional role is still under debate 13, 14.EAV N raise be used as an alternative protein candidate of diagnostic antigens and accounts for 35-40% of the total virion protein 15. B cell epitopes involved in the immune response against EAV 16. In the fork out study, we aimed to identify the nice B cell epitope using a monoclonal antibody (mAb) against EAV N protein. Our result will provide important information for developing serological diagnosis of EAV infection and understanding the antigenic structure of EAV N protein and vaccine design.Materials and methodsEthics statementCare and use of laboratory animals and all animal experiments were in accordance with animal ethics guidelines established by the Institutional puppet Ethics Committee in China. All animal studies were approved by the Animal Ethics Committee of Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences (SYXK (H) 2006-032).Cell lines and virusSP2/0 myeloma and Rabbit kidney 13 (RK-13) cells were nicetyd and keep in Dulbe ccos modified Eagles medium (DMEM Invitrogen, Carlsbad, CA, USA) in a humidified 5% CO2 atmosphere at 37C. All culture media were supplemented with 10% heat-inactivated fetal bovine serum (GIBCO, Invitrogen) and antibiotics (0.1mg/ml streptomycin and 100 IU/ml penicillin).The Bucyrus strain of EAV (GenBank accession No. NC-002532.2, a highly cell culture-adapted strain provided by the key laboratory of Tropical and Subtropical Animal Viral Diseases in Yunnan province, China) was propagated in RK-13 cells and stored at -80.Expression and characterization of recombinant EAV N proteinThe full-length sequence of EAV N gene was cloned by RT-PCR using the following primers 5-CCCGGATCCATGGCGTCAAGACGATC-3 (upstream) and 5-TTTGTCGACTTACGGCCCTGCTGGAGGCGCAAC-3 (downstream). The primers contained BamH I and Sal I restriction sites (italicized). The purified and digested PCR product was ligated into an expression vector pET32a (Novagen, Germany). The pET-N recombinant plasmid was transformed int o E. coli BL21 (DE3) and 1mM isopropyl--D-1-thiogalactopyranoside (IPTG, Invitrogen, USA) was used for inducing expression of N protein. The recombinant proteins were obtained from the bacterial lysates. The insoluble inclusion bodies were washed and solubilized with phosphate buffered saline (PBS, pH 7.4). Then, the recombinant N protein fused with 6 His-tags was evaluated by SDS-PAGE and purified by Ni affinity chromatography according to manufacturers instruction (Invitrogen).Preparation and characterization of mAbs against N proteinEAV N-reactive mAb was generated as previously described 17. Briefly, 6-week-old female BALB/c mice were immunize with the purified recombinant N protein (100g per mouse) mixed with an pertain volume of Freunds complete adjuvant (FCA, Sigma, USA). Two booster injections containing the same amount of purified N protein in an equal volume of Freunds incomplete adjuvant (FICA) were given at 2-week intervals. The purified N protein without adjuvant was injected intraperitoneally as the final immunization. afterwards three days of the final injection, the mice were euthanized and their splenocytes were fused with SP2/0 myeloma cells using polyethylene glycol (PEG4000, Sigma). The hybridoma cells were seeded into 96-well plates and selected in hypoxanthine- aminicpterin-thymidine (HAT) selection medium (DMEM containing 20% fetal bovine serum, 100g/ml streptomycin, 100IU/ml penicillin, 100mM hypoxanthine, 16mM thymidine, and 400 mM aminopterin). After 5 days, the medium was removed and replaced with hypoxanthine-thymidine (HT)-DMEM medium (DMEM containing 20% fetal bovine serum, 100g/ml streptomycin, 100IU/ml penicillin, 100mM hypoxanthine, and 16mM thymidine). After selection in HAT and HT medium, hybridoma supernatants were screened for evaluating reactivity and specificity of mAb by Western blot and immunofluorescence assay (IFA). The class and subclass of the mAb was determined using a SBA ClonotypingTM System/HRP (Southern Bio technology Associates, Inc., Birmingham, AL35260, USA).Polypeptide design and expressionEleven overlapping peptides spanning the N protein were designed (Table 1,). For each peptide, a pair of oligonucleotide strands was synthesized. Each pair of oligonucleotide strands was annealed and cloned into the BamH and Sal I sites of pMAL-C4x vector and expressed as MBP-N fusion proteins. These MBP-fused proteins were named consecutively MBP-N1 to MBP-N11. The recombinant plasmids were transformed into E.coli Rosetta (DE3) (Novagen). Each MBP-fused polypeptide was bring on by IPTG and screened by indirect ELISA. Briefly, MBP tags and purified N protein were used as negative and positive controls, respectively. Ninety-six-well microtiter plates were coated with expressed MBP-N fusion proteins at 4 overnight and occlude with 5% skim milk for 1 h at 37. After washing three times by PBST (PBS plus 0.5% Tween-20), 100 l of mAb was added to wells and incubated at 37 for 1 h. Then, the plates we re washed three times by PBST and incubated with diluted horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Abcam, UK) at 37 for 1 h. The color was developed and the reaction was stopped with 2M H2SO4. The absorbance at 450 nm was measured. All assays repeated three times and the average of the three values was calculated.Identification of the epitopesThe MBP-N-fusion proteins were identified by indirect ELISA and Western blot using the mAb. Indirect ELISA was performed as mentioned above. For Western blot, the purified MBP-N recombinant proteins were electrophoresed on SDS-PAGE, and then transferred to a nitrocellulose membrane. nonspecific antibody binding sites on the membrane were blocked with 5% skim milk in PBS overnight at 4. The membrane was washed and incubated with mAbs for 1h at 37. The membrane was incubated with HRP-conjugated goat anti-mouse IgG secondary antibody after five times washing with PBST. Following an other five times washing, the color was develop ed using 3,3-diaminobenzidine (DAB) and terminated by rinsing the membrane with deionized water.Homology analysisTo evaluate the saving of the identified linear epitope among EAV from different geographic areas, the identified epitope and the corresponding regions of other regional EAV virus strains were aligned using DNAMAN software (Lynnon BioSoft Inc., USA).ResultsProduction of recombinant EAV N protein and mAbAs shown in Fig.1a, Recombinant EAV N protein was successfully expressed in the procaryon expression system. A clear single target band with expected molecular weight was displayed. Accordingly, the recombinant EAV N protein was suitable as an antigen for immunization and hybridoma screening.Purified proteins were utilized to immunize BALB/c mice. After cell fusion and selection, an EAV N-reactive mAb generated from one hybridoma cell line was selected for its strong reactivity against N protein. This mAb was designated as 1C11. As shown in Fig.1b, c, mAb 1C11 reacted with recombinant N protein and total protein of EAV (Fig.1b, c). The reactivity of mAb was in like manner assessed using RK-13 cells by IFA (Fig.1d). The mAb only reacted with EAV infected cells and not reacted with uninfected control RK-13 cells.Identification of EAV N epitopeTo localize linear antigenic epitopes within the N protein, 11 16-amino acid long MBP fused peptides (MBP-N1 MBP-N11) were expressed and probed by mAb 1C11 by indirect ELISA. The results showed that MBP-N10 (91TVSWVPTKQIQRKVAP106) and MBP-N11 (95VPTKQIQRKVAPPAGP110) epitopes were recognized by the mAb 1C11 (OD450 1) (Fig. 2a). All the left fragments (MBP-N1-9) failed to react with the mAb. Because adjacent epitopes have 12 overlaps, we deduced that the linear epitope located in the overlapping domain of MBP-N10 and MBP-N11 (95VPTKQIQRKVAP106). To identify the minimal linear peptide epitope within this overlapping domain, a series of truncated polypeptides were expressed as MBP-fusion proteins. Ultimately, the i ndirect ELISA and Western blot showed that 101QRKVAP106 was the minimal linear epitope for the reactivity of the EAV N protein recognized by mAb 1C11 (Fig. 2c, d).Homology analysisSequence alignment was performed to evaluate the conservation of the identified epitope among different regional EAV viruses (Fig. 3). The identified epitope is conserved among all EAV isolated strains, with the exception of the ARVAC which is a modified live virus vaccine strain.Discussion subprogram location of viral protein epitopes and defining the degree of their conservation may play an important role for understanding of the antigenic structure, virus-antibody interactions. It may be very useful for vaccine design and clinical applications. In this study, the B cell epitopes of EAV N protein were identified using a mAb.Epitope mapping using mAbs has become a regent(postnominal) tool to study protein structure and provides new tools to diagnose diseases and design vaccines 18. Here, we defined one peptide epitope of EAV N protein in by using an EAV N-specific mAb. To our knowledge, epitope on the N protein of EAV has been published, unless no previous studies about 101QRKVAP106 have been reported. Starick et al. 19 produced a mAb against the N protein to detect EAV. Weiland et al. 20 used the same method to produce a mAb against the N protein of EAV and to distinguish different virus isolates from semen and tissue samples after passaging through RK-13, Vero and fetal equine kidney cells. However, the minimal epitope of these mAb was not defined precisely. Similar to the work of Starick et al. and Weiland et al. 19, 20, a mAb named 1C11 against EAV N protein was prepared by using recombinant N protein expressed in E. coli and used for identifying B-cell epitopes on EAV N protein. mAb 1C11 reacted well with EAV by WB and IFA, thus this antibody may be a useful detection tool in EAV diagnosis.mAbs are useful and effective for mapping antigenic epitopes of viral proteins. In thi s study, for epitope mapping, 11 overlapping peptides from EAV N protein were expressed with MBP tags and identified by ELISA to screen linear epitopes. The ELISA results showed that the epitope located in the sharing region of MBP-N10 (91TVSWVPTKQIQRKVAP106) and MBP-N11 (95VPTKQIQRKVAPPAGP110). Then this region (95VPTKQIQRKVAP106) was expressed, and 7 peptides with deletions were obtained to identify the precise epitope. According to the results of ELISA and Western blot, 101QRKVAP106 was considered as the minimal linear epitope of EAV protein. This result is different from the previous studies 15, 21 which stated that the precise epitope of N protein located in amino acids 1-69. This may be due to the discrepancy of the specificity and reactivity of the mAbs. Sometimes, a mAb can react with different locations of a viral protein.Sequence alignment showed that the identified epitope is very conservative among distinct regional EAV strains, but with a mutation of one amino acids on the ARVAC N protein epitope. This result suggests a slight regional difference emerged in this epitope. Therefore, it is possible to distinguish anti-Bucyrus EAV antibody from anti- ARVAC EAV antibody by using the epitope as antigen. This will be helpful in distinguishing the distinct regional EAV infection. This finding indicates that the N epitope of EAV identified in our study have a potential use in serological monitoring and differential diagnosis.In conclusion, one EAV N-specific mAb was developed and a minimal linear peptide epitope within the N protein was defined. The EAV N-specific mAb and the defined linear peptide epitope of EAV N protein may be useful for the development of specific diagnostic tools and design of vaccine.

The Criminal Black Stereotypes In Detail

The Criminal grisly Stereotypes In DetailIn this chapter, I firstly develop with a detailed presentation of the shameful male pigeonholes, then I bring into discussion the perceptions ab knocked out(p) Blacks and crime in the United States. The war on drugs of the 1980s represented a real subject of interest and I tried to present and discuss briefly about it because it was a powerful contri thoor to the typification of shepherds crooks as Black.Black Male StereotypesTo begin with, the society we live in, every ethnic group has a current stereotype that is placed on, regular if we admit it or non. Some of these stereotypes are positive, but most of them are negative. During ages, most of us hold a series of stereotypic views about certain races, g hold oners, or classes. These stereotypes are not an error of perception, but a form of social control and population often resent them or live up to them.For instance, when talking about stereotypes that society places upon some body, the most ones who ca-ca their back to the w every are the downhearted custody. They have always been seen as criminals or in the get through case as inferior species. Even today black men are depicted as uneducated, marijuana smoking men, that do nothing all day long but to commit crimes. solely, what we refuse to admit is the fact that the puritys are much violent and commit ten ms more(prenominal) crimes than a Black man.There are a few scholars I selected, that refer strictly to this matter. The first one is Katherin Russel-Brown, that refers to the stereotype as the criminalblackman, because crime and young person Black men have become synonymous in Ameri kindle culture. In her book, The Color of Crime Racial Hoaxes, snow-covered Fear, Black Protectionism, Police Harassment and different Macroaggressions (1998), she writes that black men have always been portrayed as corporeal threats. They are blamed for all that is not good.According to her, the criminalblack man is a myth and says that the stereotype enables the use of racial hoaxes, which she defines aswhen someone fabricates a crime and blames it on another person because of his race OR when an actual crime has been pull and the perpetrator falsely blames someone because of his race. (Russel, 199869)This quote describes best the society we live in, the reality that distorts peoples contrive. Here, the blames are not put only on black men, but on other ethnic groups too. Those who are in minority will always be treated like this, as scapegoats.Secondly, Linda G. Tucker came with another argument concerning Black men. In Lockstep and Dance Images of Black Men in Popular Culture (2007) she argues that the criminal black mens representation in popular culture help to bear on the realize. In her writings, she states that one of the most significant methods of criminalizing black males is the use of crime as a metaphor for race.In the context of athletics and sports, black men are consi dered to be suitable for this kind of career. They have win medals in all sports, in contrast with snowys that claim racial superiority in basketball game and American Football. What White people are afraid to admit is that the most exciting sports the ones played by black people.It is believed that sports somehow marginalizes black men from society. Sports resume advantage of the fact that some blacks dream to be on television, to show the world that they were born to be athletes. In order to fulfill their goal, they trade their faculty member future in exchange for the chance to shine in sports. But the reality is that few black men would go on to play pro sports. And even so, they have very short careers which do not last such longer.This system is based on persuasion, peer pressure and leads in the end to self distruction. Sport Institutions are designated to encourage black men not to take advantage of the opportunity of education, in exchange for athletic pursuits. One ca n play professional sports and still be an intelligent and capable member of the community in which he lives.The author of Journal of Contemporary Criminal Justice, that overly lead me throughout my study for this chapter is Kelly Welch. She is an assistant professor of sociology at Villanova University, where she teaches in the criminal Justice Program. Her research interests include race and crime, sociology of punishment, social justice, crime victims, and earthly concern polity.The scientific worldation I mentioned before, refers to the stereotyping of blacks as criminals. This image is so pervasive throughout society that criminal predator is used as a euphemism for young Black male. In American society, a prevalent representation of crime is that it is overwhelmingly committed by young Black men.Welch suggests that the familiarity many Americans have with the image of a young Black male as a violent and menacing street thug is fueled and perpetuated by typifications everyw here.(2007276)In fact, perceptions about the criminals individualism cannot be easily changed because it seems that when it comes to talk about crime, inevitably we talk about race too. As I mentioned before, Whites have long viewed criminal behavior as an inherent characteristic of Blacks and this can be traced back to the enslavement of Africans in the United States.The familiarity of crime with Blackness may have existed for some time and can be traced in the 1970s and premature 1980s. In that period of time, the image of the young Black man suffered a semnificative transformation, from a rapist into that of a criminal predator.Though Blacks have often been portrayed as physically threatening, the current Black stereotyping of criminals represents an unusual phenomenon. The evolving criminal image of Blacks appears to be a more threatening image than had been considered in the past. The general criminal tendency, now is taken for granted as a biological f fairness of African A mericans.2.2.1 Blacks and Crime Perceptions and StatisticsFor more than a century, this relationship between race and crime in the United States, has served as a topic of public controversy. Crime is on the top of the appoint of public concerns and the media often portray it in such manner that can affect our attitudes towards the minority groups from our society.Cesare Lombroso, founder of the Italian school of criminology, argued that criminal behavior was the intersection of biological factors, including race. He developed a theory according to which, some people were more civilized and others more savage. Black people were grouped plainly in the latter category, along with yellow and mixed group minorities.Lombroso believed that crime was first of all a manifestation of innate qualities and humans can be grouped as prone to crime only by analyzing their physical characteristics.For instance, slave holders from United States, began to associate African Americans with crime due to the fact that their physical features seem savage.As historians have noted, the southbound share of U.S. has had a higher rate of personnel in contrast with other parts. The rise of drug crimes and violence in the inner cities in 1970s and early 1980s, do people to associate black men with criminal predators.This biological perspective was criticized by the early 20th century scholars, including Frances Kellor, Johan Thorsten Sellin and William Du Bois, who argued that other circumstances, such as social and economic conditions, were the central factors which led to criminal behavior, regardless of race.Du Bois traced the causes of the disproportional representation of Blacks in the criminal justice system back to the emancipation of Black slaves in general and the convict leasing program in particular. He wroteThere are no reliable statistics to which one can safely appeal to measure exactly the growth of crime among the emancipated slaves. About seventy per cent of all pris oners in the South are black this, however, is in part explained by the fact that accused Negroes are still easily convicted and get long sentences, while whites still touch to escape the penalty of many crimes even among themselves. And yet allowing for all this, there can be no reasonable doubt but that there has arisen in the South since the civil war a class of black criminals, loafers, and neer-do-wells who are a menace to their fellows, both black and white. (1901147)Moving on to crime statistics in the late years, although African-Americans did not surpass the actual number of Whites in nationwide arrests, their presence in these statistics has been greater than their representation in the general public.For example, according to the survey do by the U.S. Department of Justice in 2004, Blacks composed approximately 13% of the U.S. population. But in 2002 they accounted for 38% of arrests for violent crimes and nearly 30% of arrests for property crimes. puerile arrest stat istics indicate that during the same year, Black youth accounted for approximately 43% of arrests for violent crimes and 27% of arrests for property crimes.In 2008 there were approximately 14,180 victims and 16,277 perpetrators of murder and non-negligent homicide inform by justice enforcement agencies to the FBI. The following table presents the racial demographics of murder in the United States for 2008OffendersVictimsWhite VictimsBlack VictimsOther VictimsUnknown VictimsWhite32.8%48.2%83.3%7.6%30.0%37.0%Black36.5%47.8%13.8%90.0%16.7%29.0%African Americans, constituting approximately 12% of the general population, were significantly overrepresented in the total arrests made. African Americans were also significantly overrepresented in victimization, representing 47.8% of all murder victims.Murder in White American and African American populations were overwhelmingly intraracial, with 83% of all White victims and 90% of all Black victims having been murdered by individuals of the same race.Secondly, for non-lethal violent crime, law enforcement agencies made arrests for rape, robbery and aggravated assault. The following table presents the racial demographics of these non-lethal violent crimes in the United States for 2008TotalRapeRobberyAggravated AssaultWhite58.3%65.2%41.7%63.3%Black39.4%32.2%56.7%34.2%(U.S. Department of Justice, Federal Bureau of Investigation, Criminal Justice Information Services Division. 2008)As it is showed in the table, white Americans were arrested more times than Black people in 2008, making up 58.3% of all arrests. White Americans, constituted approximately 79% of the total population, while though, African Americans constituted approximately 12% of the population, and made up 39.4% of all arrests for non-lethal violent crimes in the same year.Thirdly, regarding clerical crime, there have been arrests by enforcement agencies for forgery and counterfeiting, fraud and embezzlement. The following table presents the racial demogra phics of these white-collar crimes in the United States for 2008TotalForgery-CounterfeitingFraudEmbezzlementWhite67.4%67.7%67.5%64.5%Black30.9%30.7%30.8%33.4%(U.S. Department of Justice, Federal Bureau of Investigation, Criminal Justice Information Services Division. 2008. Retrieved 07 October 2009)White Americans were arrested again more times Blacks for these white-collar crimes in 2008, making up 67.4% of all arrests. African Americans were significantly overrepresented in forgery/counterfeiting, fraud and embezzlement, making up nearly 30.9% of all arrests.After consulting these tables of arrests made only in 2008, the Black mens image should improve because, these surveys represent the reality we are living in and not those negative narratives and images that media transmits about minorities. This is why they have such a significant influence on the belief of the society.2.2.2 War on DrugsOn Juky 14th , 1969, death chair Richard Nixon identifies drug abuse as a national threat . After presenting the juvenile drug-related arrests and the street crime that was happening during 1960s and 1967s, he called for a national anti-drug policy at both the federal and state level. Then, he popularized the term War on Drugs when first used it in 1971.The policies that his administration implemented as part of theComprehensive Drug Abuse Prevention and Control Act of 1970were a continuation of drug prohibition policies in the U.S. which traced back to the year 1914. The first U.S. law which restricted the distribution and use of certain drugs was the Harrison Narcotics Tax Actin 1914.The well-known war on drugs was a powerful contributor to the typification of criminals as Black. Crack was generally recognized as an inexpensive drug that was predominantly used by minorities.Americans were already familiar with cocaine before the war on drugs, however. Prior to break through epidemic, disintegrate cocaine was prevalent in White communities, with little acknowledgemen t from law enforcement. Black people began to use powder cocaine when this drug was affordable on the market. This contributed to the promotion of punitive policies that have hit the Black population. As Michael Tonry, among others, has noted in Malign Neglect Race, Crime and punishment in AmericaUrban black Americans have borne the brunt of the War on Drugs. They have been arrested, prosecuted, convicted, and imprisoned at increasing rates since the early 1980s, and grossly out of proportion to their numbers in the general population or among drug users. By every standard, the war has been harder on blacks than on whites. (1995105)According to crime surveys, racial and ethnic groups consume illegal drugs at approximately similar rates. Specifically, Whites account for almost 75% of the nations illegal drug users, while Blacks for about 13%. But Blacks, however, account for about 75% of the nations drug prisoners, which reveals the extreme disparity manifest in the national crackdow n on the drug problem .The sale and use of crack cocaine used by racial minorities, brought them a series of criminal penalties, which are heavier than those associated with other illegal drugs, used by Whites such as powder cocaine. This has resulted in a highly disproportionate number of Blacks who have been criminalized because of their drug use. The war on drugs may have been a war on Blacks or a war on Black drug use (Tonry, 1995).The relationship between race and crime rised various theories in the past hundred years. These theories have ranged from Lambrosos belief that certain group posses inherent criminal tendencies, to the one that is more widely accepted namely that of certain racial groups that are exposed to distress and this fact lead them to commit crimes more often. Poverty sometimes is thought to be the most prominent cause of crime.Donald Yacovone presented his interesting idea inThe regeneration of the Black Temperance Movement, 1827-1854 An Interpretation, sta ted as followsThe shift away from modern temperance and to total abstinence in 1836 marked the beginning of black liberty from white reform, although whites began to adopt teetotalism during the same period. Acceptance of total abstinence coincided with the adoption of political action as a reform measure and generally of greater militancy among blacks. By rejecting all alcohol, blacks not only sought to establish their personal integrity but they saw themselves as promoting the interests of the larger black community by offering practical and symbolic resistance to the forces of racism and slavery.This statement shows that the people of black antebellum society were genuinely interested in making a change. They believed that by initiating a temperance movement, they would gain the respect of the whites and therefore reduce acts of discrimination. The black temperance movements were established to diminish discrimination against blacks, but instead had more of an effect on the inte mperance of members of the community who were of all ethnic backgrounds.There were voluntary associations involved in determining the causes of crime, preventing crime, or reforming citizens who had committed crimes. And they played a huge role in helping the residents of the black community feel safe. Many of these voluntary associations contributed to aiding the suffering black population of Boston. These associations all-encompassing their aid to the black population and many allowed membership to black citizens. These groups all aided the process of liberating blacks and helping them to integrate into society.2.3 ConclusionThis chapter was meant to bring into discussion the association of crime with Blackness. The reputation of Blacks has been characterized by beliefs about predispositions toward criminality that can be traced back to the enslavement of Africans in the United States. I have shown that the current recognizability of the image of a young Black criminal has been t he result of various representations of crime. Contributions to this relationship that many identify between African Americans and criminality include actual pastime in crime, especially crack cocaine violations and violent offenses. Blacks do account for a disproportionate amount of crime arrests and are disproportionately convicted and incarcerated. But public estimates of Black criminality surpass the reality.

Skills and Responsibilities of a Nutritionist

Skills and Responsibilities of a alimentistMonica Suzanne B. CastroC arer Investigated Nutrition/Diet specializer partially 1 Educational and Career GoalsThere are many reasons why I chose to study upkeep. A) I will drive home opportunities in variant fields that involve advising people on what to eat, so jobs often are easier to get into by than they are in other fields. B) I will be able to work with those who are ill or need a special diet, in which case a hospital, nursing home or discourse center may be the best option for a job. C) I can become a private nutritionist, all types of people can come to me for advice on how to eat right.The nutrition industry is a field of wellnesscare focused on improving everyday health and state of well-being. I whoop it up activities often associated with nutrition and fitness including yoga, running, outdoor activities, meditation, healthy diet, organic living, and more. My goal is to further my education, get certified and become a n utrition specialist. As the health and fitness trend is sweeping the world, it is affecting and revolutionizing the health industry like never before. The demand for young, ambitious health and wellness graduates is continuously rising and a myriad of great job opportunities are opening up in different fields of nutrition science. With the evolution in this industry there is an make up scope of enjoying a successful career in this exciting field of healthcare. It is hard work, and for the dedicated, the pay can be very good to excellent. But I would do it because I want to share my knowledge and help other people, not for the money.Part 2 Employment Opportunities and Applications of Applied LearningPotential Employers chore title Nutrition SpecialistGENERAL PURPOSE Under close supervision, prepares hot and nutritious meals for homebound individuals and senior citizens.QualificationsEducation and Experience 2 (2) historic period of meal supply experience OR an equivalent combina tion of education and experience providing requisite skills to successfully perform position requirements.Special Requirements moldiness own Food Handlers Card at time of hire. Physical Demands / Work Environment Works in an industrial kitchen and senior center facility. Must be able to lift up to 50 lbs. without dish upance, greater than 50 lbs. with assistance or use of mechanical aid.Miscellaneous InformationReports To Community middle Manager Supervision Exercised NoneJob Title Nutrition Specialist/Outreach WorkerPREFERRED MINIMUM QUALIFICATIONSGraduation from utmost school or GED equivalent, with previous public contact work experience preferred. Previous supervisory experience preferred. Two years of Food Service training required.Knowledge, Skills, and Abilities(A) office to learn example Microsoft software and Food Works software.(B) Ability to obtain yearly Food Safety Sanitation Training.(C) Ability to obtain CPR and First Aide training.(D) Ability to drive a motor vehicle.(E) Ability to handle stressful situations.(F) Ability to perform duties with a high degree of attention to detail with a high degree of accuracy.(G) Ability to establish and maintain effective working relationships with Older Americans staff and the general public.Job Title Nutrition SpecialistQualificationsTwo years experience as a cook in a non-fast diet restaurant or healthcare setting.Familiarity with basic kitchen principles, portioning, stock management.High School lambskin or equivalent,Oregon Food Handlers cardServSafe Certification preferred.Experience growth and development by means of our continuous training, as well as tuition and licensure reimbursement, or pursue opportunities to advance both clinically and administratively with Youth Villages.Job Title Nutrition AssistantIn this position you willAssemble and carry through patient, visitor, and staff meal orders in a high-volume, fast-paced environment to patients rooms or designated location.Visit patients who did not place a meal order and assist them as appropriateMaintain cash bank and process cash sale ordersRetrieve meal trays from patients roomsRequiredqualifications for this position include-High school lambskin or GED- associate customer service experienceJob Title Nutrition Services SubstituteEssential job functions include the following Assist with preparation of meals according to specifications from the Cook Manager, Head Satellite Cook or Nutrition Services Specialist. Clean all food service equipment and preparation and service areas subspecies dishes, pots, pans and utensils used in preparation and service of meal. Set up serving line and gather carts, trays, napkins and utensils place items in proper location for customers. Serve food to students and staff.Other functions of the job include but are not limited to the following Operate computerized register (Point of Sale System). Attend workshops and in-services for continued professional growth and training. Other d uties as assigned.Knowledge, Skills And Abilities RequiredTo perform this job successfully, an individual moldiness be able to perform each essential duty satisfactorily. The requirements listed below are representative of the knowledge, skill, and/or ability required. Reasonable accommodations may be made to enable individuals with disabilities to perform the essential functions.Understand and follow oral and written directions. misgiving of sanitation and safety practices related to handling and serving food.Ability to use standard kitchen equipment, utensils and measurements.Basic food preparation including washing, cutting and assembling food items and ingredients. unremitting attendance.Physical Requirements Continuously stand, reach with one or both hands, lift up to 15 pounds frequently twist, lift up to 40 pounds, push and institutionalise up to 15 pounds occasionally crouch, kneel, stoop, lift up to 50 pounds.Potential Customers In My Local AreaRachel Geller She is intere sted in getting a own(prenominal) nutritionist. She is trying to eat healthier and shred some unwanted pounds.In order to accomplish her goals, I have to discuss and tackle her personalized eating and exercise plan, she will have to start off slow and eventually work her way up, to an everyday natural routine. She will have to start a daily food diary it will aid her in keeping track of her daily calorie intake. It will also aid her by presentation what recommendations she need and how much of them she need, that way she can fulfill her daily nutritional recommendations.The Skills I Have Obtained Nowadays, the requirement for academic achievement play less key role as compared to the soft skills. Competitive world has led the organizations set up their recruitment based on other non-academic factors. Knowledge is being gained through experience, not from the formulae from the prescribed syllabus. The knowledge gained from syllabus 3 years ago is not necessarily applicable in tod ays scenario. Therefore, students like me have to update ourselves with general knowledge and current issues happening in the world.I believe that in useing all of my lifelong studies, the lifelong study definitely increase my general knowledge and skills. I am more sensitive about the dynamic environment, have higher flexibility to make my future career as a nutritionist better.Part 3 Continuing Education and Career DevelopmentI want to further my education in nutrition sciences by researching and taking online programs. I believe in long life learning and self-paced studies. It will give me many opportunities to intensify my skills while pursuing my occupation. Having entranceway to origination Wide Web all over the world, with the information I need at my fingertips will propel me to a higher level of intellectual aptness. The big number of nutrition programs available over the internet enables me to arm myself with the knowledge needed to aid me on any project or endeavor th at I may attempt. Ashworth College is one of those ideal online institution that helped me pursue my dreams and ambitions to become a nutritionist. It goes above and beyond the call of duty to ensure that its students become useful members of society. As I commence my journey into real life challenges, Ashworth College will be my guide making sure that I follow the right track into the future.Two Universities I plan to apply after Ashworth CollegeKaplan University Bachelor of acquirement in Nutrition ScienceTotal Core Credits 33Total course Credits 180In this program you will explore various topics, concepts, values, research methods, and applications in nutrition science.Your coursework will focus onExploring the principles and pharmacology of nutrition and how dietary choices can contribute to a healthy lifestyleAcquiring an understanding of digestion, absorption, and metabolism of foods and nutritional needs at various stages of the life musical rhythmDeveloping the administr ative, ethical, and professional skills necessary for leadership positions throughout the health care fieldCompleting an original comprehensive capstone project that demonstrates your ability to apply your nutrition science knowledge and skills to real-world scenariosUniversity of South Dakota Bachelor in Health SciencesThis online Health Science degree program provides the opportunity for students with an interest in the health professions to explore the many opportunities in the field and to develop the knowledge, values and interpersonal skills needed to be successful in todays health and human service settings.Certifications Related to My Field of StudyThe Certification Board for Nutrition Specialists (CBNS) The clinical Nutrition Certification Board (CNCB) is a 501(c)(3) non-profitTax-exempt certification agency which provides professional training, exam and certification for health care organizations, specialty credentialing programs and state license/certification examinat ions.The Purpose of Certification The Certified Clinical Nutritionist (CCN) ExaminationEstablishes reputable standards of morality* Creation of a quality examination, based on a validated scope of practice which appropriately represents the field of clinical nutrition* Candidate access to the broad scope of clinical nutrition training and* A mechanism for re-certification every five years, recognizing continuedcompetency in the field.The CCN Examination Candidate must1. Satisfy the CORE Requirements2. accommodate Credential Review Application and College Transcripts3. Obtain Credential Review Approval4. Complete the Post Graduate Studies in Clinical NutritionProgram (PGSCN) 5. Achieve a passing score on the written CCN ExaminationThe CCN Examination processes and the examination data bank are reviewed and rewrite annually, in accordance with the specifications of the Role Delineation Study.The CNCB utilizes a criterion referenced passing point for each section of the certificatio n examination.To obtain a security system of Completion in the field of Clinical Nutrition ONLYCandidate does not meet the minimum standard in core coursework or degree(s).1. Submit your Candidate Profile Page to the CNCB.2. Complete the 56 hour PGSCN course to obtain a Certificate of Completionin the field of Clinical Nutrition through the Clinical Nutrition Certification Board.MAINTAINING THE CERTIFICATIONThe CCN Credential requires on-going maintenance to verify that the practitioner iscontinuing to update knowledge and skillsForty approved CE Hours every 2 years, fulfilled by the following1. Attendance at three out of four IAACN Annual Scientific Symposiums (60 CEs earned).2. Payment of Annual CNCB sustenance Fee. (No refunds in part or in full)3. Recertification in the spring of your fifth year via annual study textbooks with contracted Open Book Exam. Recertification Text books bestow 24 CE hours.The CCN Continuing Education Maintenance Requirement of attendance at3-out-of- every-4 IAACN Scientific Symposiums will satisfy the above statedCE requirements.Part 4 Job Application DetailsApplicants Name (last) Castro FirstMiddle Initial B. Monica SuzanneSocial Security 518-50-7839Mailing speak to (Number) bridle-path1632 N Normandie Avenue, Apt 315Work Telephone (323) 345 5839City Los Angeles Zip code 90027State CaliforniaEducationName of SchoolFT Career CollegeAshworth College military position of SchoolLos Angeles, CA6625 The Corners Parkway,Suite 500Norcross, Georgia 30092Degree or Course of StudyCertified care for AssistantNutrition, Diet and Health ScienceDate CompletedMay 2013March 2014Employment History Begin with your most recent job. List Job SeparatelyJob Title Nursing Assistant Dates worked from 2012 to PresentName of Employer Mark AuriaName of Supervisor Estelle FranciaDuties PerformedExperienced ambulatory and non- ambulatory elderly patients. Administered medication, documented food and fluid intake, measured and charted resilient si gns, prepared meals, provided personal care including feeding, grooming and dressing.PERSONAL REFERENCES List the names of references that employers may contact.1 NameNoelia BlackTelephone (323) 345 8709RelationshipInstructorAddress City State123 Easton Blvd., Los Angeles CAZip code 900782 NameApril SkyTelephone (434) 346 7563RelationshipTeacherAddress City State564 walnut tree Drive Los Angeles CAZip code 90045

Comparison Between Grapes Of W :: essays research papers

People who feel trapped often do desperate things that be active themselves and others. &8220Fifth Business and &8220The Grapes of Wrath are two novels that both lie in of trapped characters that are affected physically, emotionally, and socially. These elements are the very foundation of entrapment, which lead to the downfall of the characters. Mary from the novel &8220Fifth Business and Grampa from &8220The Grapes of Wrath are two characters that have a fatal flaw in their lives and characteristics that lead to their destruction.Mary and Grampa are the two characters that can be portrayed as being physically trapped. Mary is physically trapped by her own actions. When she meets a tramp in the gravel pit, she allows him to make love to her. This event causes her husband Amasa to resign from his job as a Baptist Minister due to shame and disgrace. After this happening Amasa keeps her tied in a harness so she cannot get out of the house. Mary&8217s actions affect and ruin her family life. Later in the story it is mentioned that she is in a small hospital behind bars and often under sedation due to her black behavior towards Dunstan Ramsay. Dunstan is the one who has cared most for her during her life however, she considers him her worst enemy for keeping her son away from her. Her physical entrapment due to her behavior and actions, eventually lead to her death. Grampa is as well as physically trapped, because of his old age. His inability to move around freely has an impact on his lifestyle. Grampa is always trapped in his household and in the vehicle the family is travel in to get to California. When he does not wish to travel, he is drugged and is forced to go on the journey. Now the entrapment can be seen in the transport he is physically unable to do anything and ultimately dies. He is not able to go anywhere and is restricted to visit places due to his old age. aroused entrapment is one of the major factors in the destruction of the characters. Mary is emotionally trapped in the past. Even years after her son ran away, she still feels that he is a little boy. Her emotional condition ultimately drives her insane. Her insanity changes her thoughts and ideas. This can be seen when she blames Dunstan for keeping her son away from her.

Azalea Seafood Gumbo Shoppe Essay -- Business Marketing Case Study, so

Azalea Seafood Gumbo ShoppeIntroductionFound in approximately 1000 supermarkets and 300 restaurants, Azalea Seafood and Gumbo Shoppe is among the largest producers of ready-to-eat gumbo with yearly revenues in 2000 of more than $1 million. The companys gumbo was their best selling item earning nearly 90% of yearly sales. Although Azalea has a competitive advantage with its tasty gumbo, Azalea is expression to maintain long-term growth and obtain a sustainable competitive advantage. In order increase its profitability, the company needs to consider options to improve its byplay strategies.Currently Azaleas facilities are located in a small country side building and many large restaurants do not find it at par with their standards. This causes Azalea to lose sales from businesses like Applebee?s and Cracker Barrel. In addition, major highways for the rapid distribution of their product are not available and slowly accessible. Despite the emplacement, Azalea?s owners are satisfied w ith the new facility and its improved capabilities, compared to that of their old location. In order to increase sales by 50% and military service Azalea to obtain a sustainable competitive advantage, the company needs to develop four strategic plans. The first one includes staying in their current location. Staying at this location will maintain their satisfaction with the facility and allow them to maximize their efficiency. Next, Azalea should implement a Total Quality Management (...

Definition Essay - Genre -- Definition Essay

Definition Essay GenreGenre, in the most generic definition, takes the meaning kind sort style (OED). Prior to the terms inception, the popular opinion of genre in the study of media emerged in The Poetics, with Aristotles discussion of the mode or manner of imitation in poetry. Of this Aristotle writes, the medium being the same, and the objects of imitation the same, the poet may imitate by narration - in which case he can either take another personality as Homer does, or speak in his have person, unchanged - or he may present all his characters as living and moving before us (Aristotle, 53). Here lies the distinction between epic, lyric, and drama, a distinction based solely in convention, the usage of the medium, independent of specific content (see also Narrative/Lyric/Drama). Yet Aristotles genre binary of disaster and Comedy rests on some observation of the objects of imitation themselves Comedy aims at representing men as worse, Tragedy as better than in actual bearing (Aristotle, 52). Later, in Section IV of The Poetics , Aristotle offers another method of generating this binary, based in historical observation (or some semblance of it). One final consideration, that of a works purpose, arises in Aristotles definition of Tragedy as through pity and fear effecting the proper purgation of these emotions, (Aristotle, 61). In this way The Poetics sketches out the basic framework of genre til now this framework remains loose, since Aristotle establishes genre in terms of both convention and historical observation, and defines genre in terms of both convention and purpose. In variety of Criticism (1957) Northrop Frye wrote, We discover that the critical theory of genre is stuck precisely where Aristotle left ... ...ohns Hopkins University Press, 1979. Frye, Northrop. Anatomy of Criticism. Princeton University Press Princeton. 1990. Lacey, Nick. Narrative and Genre Key Concepts in Media Studies. New York St. Martins Press, 2000. Liddell & Scott. An arbitrate Greek-English Lexicon (Middle Liddell). London Oxford University Press, 1968. MacLuhan, Marhsall. Understanding Media. Malmkjr, Kirsten. Genre Analysis. The Linguistics Encyclopedia. Ed. Kirsten Malmkjr. New York Routledge, 1991. Monaco, James. How to Read a Film. Oxford University Press New York. 1981. Trowse, Nadeane. The Exclusionary Potential of Genre Margery Kempes Transgressive Search for a Deniable Pulpit. The palaver and Ideology of Genre. Cresskill, NJ Hampston Press Inc., 2002. Warshow, Robert. The Immediate Experience. Harvard University Press Cambridge. 2001.

Free Bluest Eye Essays - Learning to Hate :: Bluest Eye Essays

The Bluest Eye  - Learning to Hate     Many Americans today be not meet with their physical appearance. They do not feel that they are as beautiful as the women on television or in magazines. The media is brainwashing American females that if they are not slim and have blonde whisker and blue eyes, they are not beautiful. This causes women not only to detest the ideal females, only if also hate themselves. In Toni Morrisons novel The Bluest Eye two of her main characters, Claudia and Pecola show hatred toward others, and themselves because they are not as beautiful as the supreme females.   Claudias hatred starts at the beginning of the novel when she and her sister are staring at Rosemary Villanucci. Rosemary has what Claudia and Frieda want. They want the things that snow-clad people have. We stare at her, wanting her bread, but more than that wanting to poke the arrogance out of her eyes and smash the pride of ownership that curls her chewing m outh.(Morrison, p.9) Claudia and Frieda hate Rosemary because she has both of the things that Claudia and Frieda will neer have or be, particularly Rosemarys white skin. This forces a feeling of self-hatred for being black upon the girls.   You can see Claudias hatred again when she receives a white baby doll for Christmas. Instead of adoring and cradling the new gift, as most other children would have done, she mutilated and destroyed the doll. Adults, older girls, shops, magazines, newspapers, window sign - all the world had agreed that a blue-eyed, yellow-haired, pink-skinned doll was what every girl child treasured. Here, they said, this is beautiful, and if you are on this day worthy you may have it. (Morrison p. 20-21) She hated the dolls blonde hair and blue eyes staring back at her, reminding her of how different she looked from the doll. She knew that she was wrong for destroying the doll, but she could not refrain herself from doing it. The doll, symboli zed the perfect girl, and she knew she was very far from looking the like her. In Emily Pragers essay Our Barbies, Ourselves, she reveals the damaging effect of a doll that establishes such an impossible standard of physical perfection for little girls.