Child Labour Essay Example for Free

Child Labour Essay What is child labour? According to wikipedia, Child labour refers to the employment of children at regular and sustained labour. In many countries child labour is a well known factor for employment, especially in the under developed places. Approximately 153 million children, aged between 5 and 14 are children that work in labour. Although, child labour is recognized as being children preforming work under the age of 18. There are approximately 218 million child labourers around the world. Making children do the work that adults should be doing is morally incorrect. There are many organizations that help broadcast (describe) the issue of child labour, but overall the issue is not overly recognized. The basic, easy definition of child labour is long hours of work on a full-time, regular basis, being treated abusively by the employer, either no acces or very poor access to any education. The jobs that would be considered child labour would be selling thins on the streets or working in someone elses house as a servant. Its not so much the work itself that’s bad, but how the child is treated, how many hours a day they work and whether the work prevents school attendance. Meaning that the work is not the only bad thing, it can do some good. It is the way that the child is treated that is the worse part, if the child is treated with respect and with part time hours, as well as having access to schooling, would be a great opportunity for the child. Although, that is not the case. The children are treated with poor respect, or none, which is the number one issue, they are not treated as equals. Development cooperation and poverty reduction are central aspects of Canada’s child labour strategy The number of child labourers fell by 16 percent between 2000 and 2004. Around one in three children in Sub-Saharan Africa are workers. While the percentage of child labourers is decreasing across Africa, there are actually more children in the workforce now than 10 years ago. Out of 10 working children, seven work in agriculture, two in services and one in industry. It is estimated to take $760 billion over 20 years to eliminate child labour completely. The estimated benefit in terms of better education and health is over $4 trillion, a six-to-one difference Child labour is defined as: Its not so much the work itself that’s bad, but how the child is treated, how many hours a day they work and whether the work prevents school attendance. In the worst cases, children are trapped in these situations by debts or outright slavery. Then there are extreme kinds of child labour. One type of what are called the â€Å"worst forms† of child labour is â€Å"hazardous work,† work that is very difficult and harmful to the child’s physical development. This includes anything from carrying heavy loads and using dangerous machinery to spraying pesticides and working in unclean environments. The other worst type of child labour is called â€Å"unconditional worst forms. † That means that no matter what the circumstances are, no matter how much is paid or how little the child does, it is illegal—even for adults. Every effort must be made to end this form of labour. This includes slavery, the buying and selling of a human being (called â€Å"human trafficking†), forced or bonded labour, using children in armed conflict, prostitution, pornography and involvement in drugs or any other illegal activity. there are some 250 million children between the ages of 5 and 14 years who are in economic activity in developing countries alone.? Child labour means that they toil in mines and quarries, are exposed to chemicals in agriculture, squat in crippling positions to weave rugs and carpets, and scavenge in rubbish piles. Too many are enslaved in bonded labour, isolated in domestic service, and traumatized and abused in the commercial sex trade. according to the World Bank over 70 per cent of child workers are unpaid family workers, especially in rural areas where working girls outnumber working boys. They are engaged in domestic service, agriculture, and small family enterprises, whether in rural or urban areas. This means that four in five children are not paid, or if they are paid it is much less than most legal minimum wages These children are forced to put at stake their education, their health, their normal development to adulthood, and even risk losing their lives. Child labour and the poverty cycle are intertwined. A family living in poverty cannot send their children to school. Therefore, in order to survive, children are forced to work and therefore receive little or no education. Children who grow up without an education are much more likely to remain in low-paying (and potentially dangerous) work. It also means that girls may marry young and have children young. The cycle begins again as the young family has children to support with very little income. However, child labour should not be automatically associated with children who engage in non-harmful work. Non-harmful work is part-time and does not stop children from having access to an education. It can be beneficial in the sense that is gives them an income and the ability to help to support their families. It is also important in many instances for children to gain useful social and work-related skills. Therefore, it is vitally important to not only eliminate harmful child labour, but to also support children who do have to work so that they are able to continue their studies and gain an education. Strict bans on child labour may cause more harm than good, forcing child to work in more underground and dangerous conditions than they would otherwise. These children are forced to put at stake their education, their health, their normal development to adulthood, and even risk losing their lives. Child labour and the poverty cycle are intertwined. A family living in poverty cannot send their children to school. Therefore, in order to survive, children are forced to work and therefore receive little or no education. Children who grow up without an education are much more likely to remain in low-paying (and potentially dangerous) work. It also means that girls may marry young and have children young. The cycle begins again as the young family has children to support with very little income. However, child labour should not be automatically associated with children who engage in non-harmful work. Non-harmful work is part-time and does not stop children from having access to an education. It can be beneficial in the sense that is gives them an income and the ability to help to support their families. It is also important in many instances for children to gain useful social and work-related skills. Therefore, it is vitally important to not only eliminate harmful child labour, but to also support children who do have to work so that they are able to continue their studies and gain an education. Strict bans on child labour may cause more harm than good, forcing child to work in more underground and dangerous conditions than they would otherwise.

Treating Alcoholism

Treating Alcoholism Introduction Alcoholism is defined as a disease that is persistent, progressive and often fatal. It is not a symptom of other diseases or emotional problems; it is its own disorder. Alcohol affects every part of the body even the brain which will eventually adapt to the alcohol use by becoming dependent on it after prolonged use. Genetics and environment are factors that are influenced by this disease. Drinking can reduce life expectancy by 10-12 years and next to smoking is the second most common preventable cause of death in America. The earlier a person starts drinking, the greater their chance is of developing serious illness later on. Once dependant on alcohol, its very difficult to quit. The cause of alcoholism is unknown. Just drinking gradually and consistently over time can produce alcohol dependence and cause withdrawal symptoms during periods of abstinence, but this is not the only cause of alcoholism. To develop alcohol dependence several other factors typically come into play, like genetics, culture and psychology. The signs of alcoholism or a dependence on alcohol vary from person to person and depend on how much the person drinks or how the person drinks. The definition of alcohol use and abuse ranges from moderate drinking, which is defined as equal to or less then two drinks a day for men and equal to or less then one drink a day for women. Hazardous or heavy drinking would be defined as more then 14 drinks per week or 4-5 drinks at one sitting for men or more then 7 drinks a week or 3 drinks in one setting for women or frequent intoxication in either gender. Harmful drinking is when alcohol consumption has actually caused physical or psychological harm or alcohol consumption has persisted for at least a month or has occurred consistently for a year. Alcohol abuse is used when the person either cant fulfill work or personal obligations and/or has recurrent problems with the law. Alcohol dependence is used for people that three or more alcohol related problems in a period of one year that in cludes increased amount of alcohol needed to produce an effect earlier obtained with less alcohol, experiences withdrawal symptoms or drinks to avoid withdrawal symptoms, drinks more then intended, unsuccessfully attempting to cut down or quit, gives up hobbies or leisure activities to drink. Summary Emotional and behavior problems such as depression and anxiety put people at a high risk for alcoholism and often are the reason the elderly turn to alcohol. Problem drinking in this case can be a way to self medicate. People may also use alcohol to become less inhibited in public situations that for some may be a source of great anxiety. Those that have impulsive personalities are also at a great risk for developing dependence to alcohol, due to the fact that they have low impulse control. Alcoholism affects every part of the body causing illness, cancer and with long term consumption can even cause death. Frequent, heavy drinking is associated with a higher risk of death to injury, violence and medical disorders, like pancreatitis, upper gastrointestinal bleeding, nerve damage and even impotence. As people age it takes few drinks to become intoxicated and organs are damaged by smaller amounts of alcohol then younger people. Those that require surgery also have an increased risk of postoperative complications, including infections, bleeding and decreased heart and lung functions, along with wound healing problems. If withdrawal symptoms are present can inflict further stress on the body and delay healing. Neurological or mental disorders can be caused from binge drinking which can cause memory impairment and problems thinking and concentrating. Nerve damage from severe vitamin deficiencies can impair mental function and memory and cause emotion disorders and even psychosis, like Wernicke-Korsakoff Syndrome, that causes loss of balance, confusion and memory loss and can lead to permanent brain damage and even death. Diagnosing alcoholism can be hard, since nearly always people deny the problem. But for most, denial may be the first warning sign that their drinking is out of hand. There are tests to screen for alcoholism, most are short and allow the person to take them on his or her own time. Because people deny their problem or attempt to hide it, the questions relate to problems associated with drinking, rather then the amount of liquor consumed. The quickest test is the CAGE test and is an acronym for: 1. Attempts to CUT (C) down on drinking. 2. ANNOYANCE (A) with criticisms about drinking. 3. GUILT (G) about drinking. 4. Use of alcohol as an EYE-OPENER (E) in the morning. This test is called a Self Administered Alcoholism Screening Test (SAAST) and appears to be the most useful in detecting alcoholism in white middle aged males. Alcoholism, hard to detect in elderly women is sometimes diagnosed as depression and prescribed anti-anxiety drugs that can have dangerous effects when mixed with al cohol conclusion Treatments for alcoholism vary, but the overall goal in treatment is total abstinence, since those that abstain have better success rates then those that dont. Treatment should also, include replacing addictive patterns with satisfying, time filling, behaviors which are able to fill the void in daily activities when drinking has stopped. Because alcoholism is so difficult to treat, most doctors will choose to treat alcoholism as a chronic disease that include relapses and remission periods. Inpatient and outpatient treatments are available to those that would benefit most from these types of treatments. Those with co-existing medical or mental disorders or those that might harm themselves have greater success with inpatient treatments at a psychiatric hospital or alcohol center. Outpatient treatments work best with people that have a good support system and are able to take medications for mild to moderate withdrawal symptoms. Psychotherapy treatments focus on Psychotherapeutic approaches and include cognitive-behavior therapy is used for severe alcoholism and gives people the opportunity to learn to cope and control their behavior, by changing the way they think about drinking and Interactional group psychotherapy that includes group based therapy like Alcoholics Anonymous or AA, 12-step program. Alcoholism is a very real and serious disease that requires medical treatment and those suffering will require lifelong care and support.

Pseudomonas Aeruginosa Gene Expression

Pseudomonas Aeruginosa Gene Expression Pseudomonas aeruginosa is the gram negative rod shaped bacteria is found in all natural and man made environments. the organism is so stubburn so that it can grow in any environment and also at high temperatures such as 420C. The unique ability of the organism is forming biofilm. The biofilms are formed by the organism with the association with a surface. The surface may be anything such as cloth, paper, glass etc and any artificial surface. It is the opportunistic pathogen which behaves neutral to the normal tissues but it infects once the defences of them are compromised. The development of the biofilm starts with the attachment of the freely moving planktonic cell to a surface. Generally the twitching motility of the P.aeruginosa helps to use its unipolar flagellum to adhere a surface. The bacterium develops colonies known as microcolonies and adhere to the surface strongly which are known as pili structures (Baron S et al.,1996). During the biofilm development several factors pla ys a key role mainly the exopolymeric matrix. The EPS matrix is not visualised directly, but it can be done by using some fluorescent dyes. The functioning of the EPS matrix in holding cells of Pseudomonas aeruginosa together and forming the mature biofilms. The biofilm develops with cell division and recruitment. The cells interacts via quorum sensing and these interactions are studied under scanning electron microscopy. The biofilm formation takes place in stages and many changes are observed in its size and shape during the development. The EPS matrix is made up of polysaccharides, proteins and nucleic acids. The exopolysaccharide matrix is the important component of the biofilm. The exopolysaccharide genes such as pilB, pslA, lecA, ureB and alginate genes are responsible for the formation of pseudomonas biofilms. The locus of the psl polysacchide genes encode the cells to adhere to a surface and thus maintaining the biofilm structure. In this research project we try to quantify the gene expression of the above genes pilB, pslA, lecA, ureB using rpoD as the house keeping gene. The biofilms are grow invitro on microcarriers. The microcarriers made up of dextrons in the range of 60-90 µm in size are used as a surface for Pseudomonas aeruginosa to form biofilms. The microcarriers are spherical in size where the bacteria can easily adhere on surface. In this present work we try to grow planktonic cultures and biofilms cultures on the microcarriers in a fermentor separately. The genes responsible for the biofilm formation are quantified by comparing the planktonic and biofilm samples. The planktonic cells grow very rapidly whereas the bacterial adhering takes some time to form biofilm in other case. The samples are collected at regular intervals and mRNA is extracted. The gene expression is quantified using real time PCR. The relative quantification method of qPCR or real time PCR method does not require any standard curve for the analysis where the samples are normalised by using an internal control. The threshold CT values obtained from the curve are used to calculate relative quantification by comparative à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  CT method. The results from the planktonic and biofilm gene expression results are compared. On the other hand we also assay the nutrients such as glucose, ammonia, phosphorus and protease during the fermentation. The concentration levels of the nutrients are also compared for both biofilm and planktonic fermentation. Review of literature: Pseudomonas aeruginosa is a member of gamma proteobacteria class of bacteria. The bacteria is gram negative, rod shaped and aerobic belongs to the bacterial family of pseudomonadanceae. The recent developments in the taxonomy based on the 16s ribosomal RNA or conserved macromolecules, the family members are divided into eight groups. Pseudomonas aeruginosa is a species present in this group sharing the with other twelve members. Pseudomonas aeruginosa is a free-living bacterium, commonly found in soil, water, skin flora and in all man made environments. The organism regularly occurs on the surfaces of the plants and animals. Pseudomonas aeruginosa is an occasional pathogen to plants. It is also an increasingly emerging pathogen of clinical relevance. The organism not thrives in the normal atmosphere but also in the places with little oxygen and thus forms the colonies in natural and artificial environments. Pseudomonas aeruginosa is an opportunistic pathogen for humans, which means that it exploits some break in the host defences to initiate an infection. The bacteria never infects the uncompromised tissues, but it can infect any if the tissue defences are comprised. It can cause infections such as urinary tract, respiratory system infection, soft tissue infection dermatitis, gastro intestinal infections. It causes a variety of systemic infections in patients particularly with severe burns and immune suppressed in cases of cancer, AIDS, cystic fibrosis in lungs. Characteristics Pseudomonas aeruginosa is rod shaped gram negative bacteria. The size of it ranges between 0.5 to 0.8  µm by 1.5 to 3.0  µm. almost all the strains of Pseudomonas are motile with a unipolar flagellum. It is ubiquitous in soil ,water and on the surfaces in contact with water or soil. It has respiratory metabolism and can grow in the absence of oxygen if NO3 is available as respiratory acceptor for electron. Generally the bacterium is found in nature in the form of biofilm attached to a surface or substrate, or in the planktonic form as a unicellular organism swimming actively with its unipolar flagellum. It is one of the most vigorous and fast swimming organism found in pond waters. It can grow at temperatures at 42OC or high, but the optimum temperature is 30 to 37OC. it is resistible to a wide variety of physical conditions such as temperature and can even resist the salts, dyes, antiseptics and most commonly used antibiotics. Pseudomonas aeruginosa produces different types of colonies from the source they have isolated. They appear like small and rough colonies. Moreover the samples collected from the clinical sources look like a fried egg like structure which is smooth, large with elevated appearance. The other type of colonies obtained from the respiratory and urinary tract secretions looks like mucoid.this mucoid appearance is attributes for the production of alginate slime. These smooth, mucous colonies presumably plays an important part in the colonization and virulence. Pseudomonas aeruginosa produces two types of soluble pigments. Pyocyanin which is a blue-green and a yellow-green and fluorescent pigment called as pyoverdin. Pyocyanin is produced abundantly in the media with low levels of iron and also functions for the iron metabolism of the organism. Many strains of Pseudomonas aeruginosa produces pyocyanin but not all the strains does it. Although the organism is classified as the aerobic, sometimes the organism is considered as facultative anaerobe where it can adapt and proliferate the conditions with less or total O2 depletion. The organism can grow in the absence of oxygen if NO3 is available as respiratory acceptor for electron. It can ferment arginine by substrate level phosphorylation where the layers of alginate present surrounding the mucoid cells of the bacteria controls the diffusion of oxygen(Collins FM, 1955), this is witnessed in the cystic fibrosis of the lung infection. Pseudomonas aeruginosa which is found in the biofilm form is responsible for a variety of activities involved which are natural and artificial. Resistance to antibiotics Pseudomonas aeruginosa is opportunistic pathogen, has a notorious resistance to antibiotics. The organism is very resistance towards many antibiotics due to the potential permeability barrier provided by the gram negative membrane. Pseudomonas aeruginosa in the biofilm form makes the cells impervious to the concentrations of the therapeutic antibiotic. The organism gets associated with others like actinomycytes, moulds and bacillus because of the natural environment they live. Moreover the organism have antibiotic resistance plasmids R factors and RTFs, the genes are able transferred by conjugation and transduction following the mechanism horizontal gene transfer(HGT) (Todar.K,2008). Biofilms Biofilms are defined as a matrix enclosed bacterial populations which are adherent to each surfaces or interfaces. Microbial aggregates and floccules and also adherent populations within the pore spaces of the media are spoken in the definition (Costerton J.W, et al 1995).bacterial biofilms came into existence and became significant from their first recognition in their ubiquity. The past researches show that profoundly the growth phase of the biofilms is distinct from the planktonic. In response to the surfaces proximity, bacterial cells alter their phenotypes (Costerton J.W, et al, 1995). The early stages of biofilm formation the bacteria is in stable juxta position with those of same and other species. The micro environmental conditions, cellular juxtapositions and the extra cellular polysaccharide matrix constitute to the development of the bacterial biofilm. Different bacteria responds to the specific environmental conditions with certain growth patterns, and a structurally comp lex and mature biofilm develops(Hamilton W.A et al, 1987). Physiological cooperation is the major criteria in shaping structure and juxta positions establishment to make biofilms adherent for the surfaces. It has been proposed that large suites of genes are expressed differentially regulated. Formation The formation of the biofilm initially starts with the attachment of the freely floating bacterial cells to a surface. The bacterial colonies adhere to the surface with weak vander wall forces. These colonies can lead themselves to more adhesion called as pili structures (Baron S et al.,1996). The organism adheres more to the available surface by binary fission, exopolymeric matrix is produced to develop mature biofilms. The matrix produced holds the biofilm together. Biofilm develops with cell division and recruitment, the cell in the biofilm communicate via quorum sensing. We can observe changes in shape and size in the final stages of biofilm formation. Distribution and ubiquity of biofilms The general phenotypic strategy of organism is to change in the mode of growth that to adopt to the altering growth conditions. The cells of Pseudomonas aeruginosa which is a ubiquitous bacterial species respond to all favourable nutrient conditions. The organism adheres to the available surface by binary fission, exopolymeric matrix is produced to develop mature biofilms. The rod shaped bacteria grows in matrix which is enclosed in a sessile growth mode. The cells are protected from the biological, antibacterial, chemical agents and adverse environmental conditions. The periodically shed planktonic cells from the biofilm provide and sacrifice the protection to the new fresh habitats can colonize with the biofilm. The adherent biofilm populations has a significant metabolic activities( Fletcher.M, 1986). The extent of biofilm formation is predicted in the particular system is based on the following principles: Metabolically active bacteria show tendency towards the surfaces for adhesion in all natural environments. In all environmental conditions the extent of biofilm formation is controlled by the available nutrient for cell respiration and exopolysaccharide production. The organic nutrients sometimes tend to associate with the surfaces, in such cases the local biofilm is difficult to trigger. This is more in extremely oligotrophical environments, but bacteria normally does not adhere to surfaces in nutrient less environments. Pseudomonas aeruginosa Biofilms The communities of microbial bacteria(Pseudomonas aeruginosa) forms a definite structures with the association with surfaces are abundantly available environment. These structures are often called as biofilms which are less susceptible to antimicrobial agents. These biofilms causes chronic infections and very difficult to remove. The extracellular polymeric matrix(EPS) are known to holds the biofilm structure and it performs as a matrix. The EPS matrix holds the biofilm cells together and protects them from shear forces in fludic environments. The extra cellular matrix for Pseudomonas aeruginosa forms the biofilm development in different stages. The matrix holds the cells together on the surface and forms a critical structural design resists to many environmental changes. Pseudomonas aeruginosa biofilm develops in a five stage multicellular cycles, the process is initiated by the attachment of planktonic (free) cells to a surface. This is followed by the formation of microcolonies. T he microcolonies are seeded and gets dispersed where the cells form the microcolonies comes out to occupy a new surface( Ma. L et al, 2009;Stoodley.P et al,2002). The EPS of Pseudomonas aeruginosa composed of polysaccharides, proteins and nucleic acids. It is the important component of the biofilms. The exo polysaccharide genes lec A , Pil B, Psl A, alginate are responsible for the formation of Pseudomonas biofilms. The locus of the Psl exopolysaccharide encodes bacterial cells to surface and thus maintaining the biofilm structure. The matrix which is anchored by the Psl is not visualised directly during the developmental stages of Pseudomonas aeruginosa biofilm. In addition to the present scenario of exopolysaccharides, extracellular DNA also plays an important part in . P.aeruginosa biofilm matrix (Allensen-Holm M et al, 2006). The extracellular DNA is derived from the chromosomes and it functions for signalling between the cells interconnecting the biofilm matrix. In the biofilm matrix, extracellular cellular DNA promotes cation gradients, release of genomic DNA and induces antibiotic resistance ( Mulcahy H, et al. 2008). Real time PCR Polmerase chain reaction or simply PCR is introduced by Kary mulis in 1980, which really a revolution in the molecular biology. PCR is the standard procedure to amplify small quantities of DNA molecule into large quantities. The method depends on thermal cycling which involves the continuous heating and cooling of reaction for DNA melting and enzymatic replication of DNA. PCR applications employ a general heat stable DNA polymerase such as Taq polymerase enzyme isolated from the bacterium Thermus aquaticus. PCR method is involves three major steps.1) Denaturation 2) Annealing 3) Elongation. In the first step DNA template is denaturated to single strand at high temperature of 940C. In the second step, single stranded DNA template annealed with primers, the temperature is 650C. At 720 C the DNA starts elongation which is third step. DNA polymerase synthesizes new complementary strand to the template DNA strand. Real time polymerase chain reaction or qualitative polymerase chain reaction is a laboratory technique which not only amplifies but also quantifies the target DNA molecule. it helps the user for detection and quantification(absolute and relative) of many specific genes in a present in a given DNA sample. The general principle of polymerase chain reaction (PCR) is also used in the RT PCR. Unlike PCR, RT PCR is a new technology product is detected at the end of the reaction. There are two common methods for the product detection in RT PCR 1) Non -specific fluorescent dyes which interacts with the double stranded DNA molecule 2) Sequence specific DNA probes consists of oligonucleotides that are fluorescently labelled with reporter. The product is detected after the hybridization of the probe and complementary strand. The RT PCR is used to amplify the reversly transcribed cDNA products from the mRNA, which is becoming the effective and powerful tool in the field of molecular biology to quantify and study the gene expression. The RT PCR method is very easy to apply provides the reliable, accuracy and rapid gene expression quantification. The accurate nucleic acid quantification requires mathematical application for data analysis(Michael W.P., 2001). The real time PCR provides the very accurate and reproducible quantification of gene copies (Heid C.A., 1996). Unlike other PCR methods, RT PCR does not require post process sample handling, prevents product contamination, provides faster and accurate assay results. Real time PCR is very accurate and less labour usage than the present gene quantification methods. Nucleic acid sequence quantification has an important role in the biological research. Measuring the gene expression have been extensively used in the biological responses to various external s timuli( Tan et al, 1994; Huang et al,1995). Gene and genome quatification also used for HIV demonstrates the change in the expression level of virus in the disease period( Conner et al. 1993); (piatak et al, 1993). There are two types of quantification methods used in real time PCR: Absolute quantification Relative quantification Absolute quantification determines the input copy number generally by relating the PCR signal to a standard curve. In this method equal input amounts of sample, each sample under analysis amplifies with identical efficiency up to the point of qualitative analysis( Heid et al,1996). We quantitate the unknowns based on a known standard quantity. The unknown samples are compared to the known by creating a standard curve. The input copy number of the PCR signal is related to standard curve in absolute quantification method. Sometimes it is not necessary to know the copy number but it is important to calculate the relative change gene expression (Livak.K.J et al, 2001). The second method is relative quantification, we analyse the changes in gene expression in a given sample relative to another reference sample(untreated control sample). Relative standard again consists of two types of methods they are relative standard curve and comparative Ct method. In the relative quantification the PCR signal is related to the targeted transcript of the samples in the group. The new applications of comparative Ct method is the 2-à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  Ct method which is efficiently useful for the analysis of qPCR data. The 2-à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  Ct method(Livak.K.J et al , 2001): Derivation of 2-à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  Ct method: The equation for the amplification of PCR can be written as: Xn=X0 (1+Ex)n(1) Where Xn is the number of target molecules at cycle n of the reaction. X0 is the initial number opf target molecules. Ex is the efficiency of the target amplicon. n is the number of cycles CT is the threshold cycle, indicates the fractional cycle at which the amount of amplified target reaches a fixed threshold. XT =X0 (1+Ex)CT.X=Kx(2) Where XT is the threshold number of target molecules. CTX is the threshold cycle for target molecule and Kx is constant. A similar reaction for the internal control gene is RT =R0 (1+ER)CTR=KR-(3) Where RT is the threshold number of reference molecules R0 is the initial number of reference molecules ER is the efficiency of the reference molecule. CT.R is the threshold cycle for reference amplicon. KR is a constant. Dividing XT and RT, we get , (XT/ RT )= (X0 (1+Ex)CT.X) / (R0 (1+ER)CT.R )= (Kx / KR ) = K-(4) For real time amplification using Taqman probes the exact values of the XT and RT depend upon the number of factors including the reporter dye and fluorescence properties of the probes, efficiency and purity of the probe, florescence threshold setting. Thus k cannot be equal to one. Assume the efficiencies of target and the reference as equal. ER = Ex=E (X0 / R0) (1+ E) CT.X- CT.R =K(5) XN (1+E)-à ¢Ã‹â€ Ã¢â‚¬  CT =K-(6) Where XN is equal to the normalised amount of target gene (X0 / R0) and à ¢Ã‹â€ Ã¢â‚¬  CT is the difference of the threshold cycles of target and reference (CT.X- CT.R ). The expression can be written as XN=K(1+E) -à ¢Ã‹â€ Ã¢â‚¬  CT -(7) If we divide the whole equation by XN for any sample q by XN as the calibrator(cb) XN.q /XN.cb ={[ K(1+E) -à ¢Ã‹â€ Ã¢â‚¬  CTq ]/[ K(1+E) -à ¢Ã‹â€ Ã¢â‚¬  CTcb ]} =(1+E) -à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  CT Here -à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  CT = -(à ¢Ã‹â€ Ã¢â‚¬  CTq -à ¢Ã‹â€ Ã¢â‚¬  CT.cb) For amplicons which are designed less than 150 bp, primer and magnesium concentration are optimised, the efficiency is nearly equal to 1. Therefore the amount of target gene, normalized to an endogenous reference which is relative to a calibrator, taken as Amount of target =2-à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  CT . When the threshold is set for n standard deviations in the plot , above the mean of the base line emission from cycles 0 to n. The standard deviations is calculated from the data attained from base line of the amplification plot. When the fluorescence exceeds the threshold limit, CT values are determined at that point. The CT values are ten times more than the standard deviation of base line. The point at which amplification plot crosses the threshold can be defined as CT (Heid et al, 1996). Materials and methods Identifying pseudomonas aeruginosa Gram straining: Prepare a small amount of smear on a clear glass slide. Heat fix the smear to slide. Add a drop of crystal violet on to smear allow for 60 seconds. Rinse the slide with water. Add a drop of grams iodine to the smear allow for 60 seconds. Rinse the slide with water. Add 95% absolute ethanol to destain the crystal violet. Rinse again with water. Add a drop of saffronin over the smear, allow for 60 seconds. Rinse with water. Blot dry the slide, do not rub. Observe the smear under microscope. 10. If the colour of the bacteria appears pink it is gram negative, it is gram positive if it appears purple. Rod shaped pink coloured bacteria is clearly observed for gram negative pseudomonas aeruginosa. Shake flask method Prepare media of nutrient broth about 13 grams per litre of distilled water. Mix well and pour 50 ml of media into a 250ml conical flask. Plug the conical flask and wrap the mouth with aluminium foil. The less volumes of media in the flask increases surface area and oxygen transfer rate(OTR). The flasks are autoclaved at 1210 C and 1.5 lbs pressure for 1 hour 15 minutes. Planktonic-contains 50 ml of nutrient broth Biofilm-contains 25ml of PBS with microcarriers plus 25 ml of nutrient broth. Inoculate a loopful of pseudomonas aeruginosa into 50 ml nutrient broth aseptically. Allow the bacteria to grow overnight. Take the optical density at 550nm using spectrophotometer.The amount of the inoculums to be added to the shake flasks for analysis is calculated using the formula below: Amount of sample to add into shake flask = [working concentration X volume(ml)] / stock(OD) obtained After inoculation the flasks are kept in the orbital shaker at 300C. The samples of 1 ml is collected for RNA extraction at particular time intervals at 0,1,2, 4,24 and 48 hours for planktonic. In respect the biofilm samples are collected at time 0,22,24,26,44,46 and 48 hours as their growth is slower than the freely suspended planktonic cells. Then 2 ml of RNA later solution is added to the collected samples for RNA stabilization. The optical density is regularly monitored to check the growth of pseudomonas aeruginosa . Fermentation The fermentation process we perform here is batch fermentation. The bioreactor used for the study is a small scale fermentor from APPLICON BIOTECHNOLOGIES. The process will take place in a 3L fermentor(consists the working volume of 2.5L) for about 24 hours. During the run the criteria of temperature is kept at 300C . The pH is run free, stirrer speed is set at 600rpm for planktonic culture. The fermentor is run free for 24 hours overnight to check the production of foam. The cold finger helps to control the temperature by feeding water from/to the reactor. The o ring is checked for any leakage. The sample port is a tube for sampling using back pressure and forces the liquid into a glass recipient. The inlets for acid, alkali and antifoam are linked to durham bottles, going through the pumps. The durham bottles are blank, because above parameters are not controlled in this process. Air is pumped through a filter. The condenser is connected to water that comes out of the fermentor. Wh en the water is warmer than outside, condensed water reaches the condenser. The gas goes from an outlet connected to the condenser. The dissolved oxygen probe is connected to dO2 cable from the machine. It measures the amount of dO2 in the media. This probe needs to be kept in the buffer, when not in use. The probe should be plugged for 10 to 12 hours before the inoculation inorder to polarize. The stirrer is placed on the top of fermentor. The vessel is heated with the help of heating pad and temperature is detected using temperature probe. The air inlet is set up through the pumps and its volume is controlled with a rotometer in front of the machine. In this case 2.5L of air/min is supplied to the fermentor. For the gas analysis of oxygen and carbon dioxide the gas is passed through a drying column and reaches the monitor which is already set up for calibration. we run two runs of fermentor for both planktonic and biofilm cultures. The parameters such as temperature, aeration for planktonic and biofilm cultures are almost same. The only parameter varies for both is stirring is maintained at 600rpm for planktoni c and 200rpm for biofilm fermentation. Procedure: The fermentor is sterilised and all the valves are sealed with foil before going into the autoclave. The fermentor is run for about 24 hours before adding the inoculum to check any formation of foam. About 20 ml of an overnight culture is added to the fermentor. Sampling :About 7ml of sample is collected for every 20min for planktonic and every 2 hours for biofilm until the exponential phase is reached. The sample collected is preserved in the freezer for different analysis. The sample is assayed for gene expression by adding 2 ml of RNA later solution for RNA stability. The samples are assayed for ammonia, phosphorus, glucose and protease. The fermentation conditions are listed below. RNA EXTRACTION (protocol provided by applied biosystems) Materials required: 2-mercapto ethanol,100% ethanol, 10% SDS(in RNase free water), 0.5  µl/sample. Lysozyme solution: 100  µl/ sample 10mM Tris HCl (PH 8.0)mM EDTA 10mg/ml lysozyme (in RNase free water) Lysis buffer: Before beginning the lysis and homogenization steps, prepare a fresh amount of lysis buffer containing 1% 2-mercapto ethanol for each purification procedure. Add 10 µl 2-mercaptoethanol for each 1ml lysis buffer. Use 350  µl of freshly prepared lysis buffer for 1109 bacterial cells. Lysis and homogenization: Harvest 1109 bacterial cells and transfer them to an appropriately sized microcentrifuge tubes. Centrifuge at 500xg for 5 minutes at 40C to pellet cells. Discard the supernatant. Add 100  µl of prepared lysozyme solution to the cell pellet and resuspend by vortexing. Add 0.5  µl 10% SDS solution vortex to mix well. Incubate the cells in the tube for 5 minutes at room temperature. Add 350  µl lysis buffer prepared with 2-mercaptoethanol. Vortex to mix well. Transfer the lysate to a homogenizer inserted in an RNase free tube and centrifuge at 12,000xg for 2 minutes at room temperature. Remove the homogenizer when done. Purifying the RNA from bacterial cells: These steps are followed to bind wash and elute the RNA from the P.aeruginosa cells sample. Add 250  µl 100% ethanol to each volume of bacterial cell homogenate. Mix thoroughly by vortexing to dispersing any visible precipitate that may form after adding ethanol. Transfer the sample(including any remaining precipitate) to a spin cartridge( with a collection tube. Centrifuge both the spin cartridge and collection tube at 12,000xg for 15 seconds at room temperature. Discard the flow through and re-insert the spin cartridge in the same collection tube. Add 700  µl of wash buffer I to the spin cartridge. Centrifuge at 12,000xg for 15 seconds at room temperature. Discard the flow through and the collection tube. Place the spin cartridge into a new collection tube. Add 500  µl wash buffer II with ethanol to the spin cartridge. Centrifuge at 12,000Xg for 15 seconds at room temperature. Discard the flow through and re insert the spin cartridge into the same collection tube. The steps are repeated once again. Centrifuge the spin cartridge and collection tube at 12,000Xg for 1 minute at room temperature to dry the membrane with attached RNA. Discard the collection tube and insert the spin cartridge into a recovery tube. Add 50  µl of RNase free water to the centre of spin cartridge. Incubate at room temperature for 1 minute. Centrifuge the spin cartridge and recovery tube for 2 minutes at 12,000Xg at room temperature. Store the purified RNA or proceeded for DNase treatment after RNA treatment. (store purified RNA at -800C for long term). Agarose gel electrophoresis: The RNA samples after extraction are checked for the RNA signal. To visualise the RNA 1% agarose gel electrophoresis. The materials required for agarose gel electrophoresis is TAE buffer and agarose. TAE buffer(50x Tris acetate) 242 g of Tris base 57.1 ml glaciel acetic acid 100 ml of EDTA 0.5M PH .8.0 700ml of dH2O . About 20 ml of above solution is mixed with 980ml of distilled water to make it 1X. 1 gram of agarose is added to 100 ml of TAE buffer. The solution is mixed well. The solution is heated until the agarose is completely dissolved and solution becomes colourless. About 2.5  µl of ethidium bromide is added to the solution to visualise the RNA bands. Special care is taken while adding the ethidium bromide because it is bio-hazardous. Then about 5  µl RNA sample is mixed with 1  µl of DNA loading dye. The RNA samples are RUN with the potential difference of 80V with DNA Hyper ladder II as a standard. DNase treatment (protocol and kit provided by applied biosystem) The DNase I treatment is performed to remove any traces of genomic DNA from the eluted RNA. Protocol: Add 10x DNase buffer and DNase I to the RNA sample in the following composition: Amount component 1/9 th volume 10x DNase buffer 4  µl DNase I The components are mixed with gentle pippeting. Then the mixture is incubated at 370C for 90 minutes. DNase inactivation reagent is added to the solution and mixed well, leave at room temperature for 2 minutes. The samples are centrifuged at maximum speed for 1 minute to pellet the DNase inactivation reagent, after that the RNA solution is transferred to a new RNase free tube. The samples are run on 1% agarose gel electrophoresis to check the DNA contamination. We fou

Seminole Indians :: essays papers

Seminole Indians Effects on European Settlers and The United States. Timeline of Seminole History 1528-Spain lands in Florida for first time (Cabeza de Vaca), 1817-First Seminole War. 1819-Spain cedes Florida to the United States. 1832-Second Seminole War begins. 1835-Osceola is captured by Andrew Jackson. 1838-Osceola dies in Federal Prison. 1842-Second Seminole War ends. 1845-Third Seminole War. 1907-Oklahoma Enters the union with most of the residents from the southeast part of The United States. In June of 1528 Cabeza de Vaca and the Spanish came to what is now known as Florida for the first of many trips. They came capturing the women and children along with one governor of the Seminole Tribe. The Seminole asked for their tribe member’s back. Spain gave them the women and children but not the governor. Spain responded by attacking and chasing the Indians across the swamps killing only one Indian. The Spanish attacked once again and only killed one Indian again. The Spanish fled to their boats escaping Florida. The Seminole had mounted their first of many defenses against European Settlers that would wreak havoc on The United States of America in the 19th century. Who are these light skinned people who come tour land They arrest our chief and take away our women and children We ask for our people back, and they gave us our women and children, but not our chief Then they attack us, not defeating us; they flee in terror, killing one man They say our land and our people are poor They attacked us once more but our strong warriors held up They fled to their ships and departed These light skinned men were foolish to try to push us around on our land They killed two of our tribe members and a Spanish commander in battle This poem was called Foolish White People and was inspired by Alvarez Nunez Cabeza de Vaca from Relation of Alvarez Nunez Cabeza de Vaca. Seminole is a word that means "wild" or "those who camp at distances." It is equal to the American word "frontiersman." The Seminole were a tribe that broke off from the Creek civilization of Georgia. The Seminole were from the southeast region of the United States. The tribe lived in houses called chickees (see drawing one) When white people came to colonize, as time went on, they referred to the Seminole tribe as part of the "Five Civilized Tribes." The other four tribes were the Choctaws, the Chickasaws, the Creek, and the Cherokee.

Legal Progression of Marriage in America :: Essays Papers

Legal Progression of Marriage in America As the British colonies in North America took root, a great part of the economic growth in colonial society was predicated on the labor of imported African slaves. As the number of slaves increased significantly, especially in the southern colonies, a system of separation of the races was established. Since the beginning of the White and Black coexistence in America, Whites have suppressed the rights of Blacks in order to emphasize their racial superiority and to prohibit mixing between the races. In order to maintain their racial purity, Whites established laws making illegal the marriage of Blacks and Whites. Although anti-miscegenation laws were present in early colonial societies, the legacies have continued in the contemporary period. For example, Alabama amended its constitution in 2000 to acknowledge interracial marriages as valid and legal. For over 300 years, anti-miscegenation laws have remained generally the same, outlawing marriages between people of different races. Over time, however, definitions of who is Black have drastically changed, reflecting the status of Blacks’ in society. Anti-miscegenation laws during the era of slavery defined Blacks as having at least one Black grandparent, or one quarter â€Å"Black blood†. Later, these same laws during the Great Migration of the twentieth century, evolved to define Blacks by the â€Å"One Drop Rule,† a rule stating that one was Black if he or she had at least one Black ancestor. These laws, as represented in this paper and based on the policies of the colony and state of Virginia, were used to maintain a separation of races and, thus, to preserve the purity of Whites and to guarantee their system of White supremacy. Sent by King James I, The Virginia Company established the first colony in America in 1607 appropriately named Jamestown (History of Jamestown 1). It is generally accepted that the first Blacks were imported to America in 1619, only twelve years after the colony was established. At this point, no specific laws prohibited interracial relations, but societal taboos and religious doctrines were enough to separate Whites and Blacks from sexual contact. Such beliefs were noted in the proceedings between the governor and his council within the colony: In 1630, a man named Hugh Davis, accused of being sexual involved with a Black, was â€Å"to be soundly whipped before an assembly of Negroes and others for abusing himself to the dishonor of God and the shame of Christians by defiling his body and lying with a Negro† (Hall 602).

Romulus My Father Notes for Belonging

* Study Questions * * Chapter 1 * 1. Romulus has a crisis in identity. He feels he is accepted in society due to his skill in blacksmithing trade. Although accepted, he does not feel he belongs due to his culture and heritage. His character is tied up with a strong moral code, in working hard, pride, communal strength and contributing to society but has no confidence in identity and how he fits in. He â€Å"Always considered himself a Romanian. † And felt he belongs to that part of his heritage. He carried this culture through to Australia and keeps these character traits, which could be the reason he is separated socially.He is a religious man that takes education seriously and in the highest regards. He always knew poverty and hard work for the community rather than money, which is why he was able to work so hard as a blacksmith for no pay. This shows his strength of character and good work ethic. He felt attached to Europe and therefore detached from Australia. In retrospec t he was accepted by many people due to his hard work but still didn’t ‘belong’. * 2. Retrospective, hard working before and after school. Romulus prides himself on character.The luxury that he didn’t get his son now is able to enjoy. Also not having a childhood because he worked the whole time. Raimond was able to become a writer and enjoy more creative things in life. He had free time where as his father didn’t. His dad grew up with little money and had to work for all the money he needed where as Raimond didn’t have to work as much. Most importantly however, Raimond was able to enjoy a sense of belonging when he was growing up to his father and family. Romulus was unable to enjoy any real sense of belonging whilst growing up.Raymond would not have the same work ethic as his father; His dad thought that everything would revolve around his work. He worked before and after school. Romulus is a pragmatist or a practical man due to his job to g et things done. This is because of his lack of childhood he has no humor, etc. Raimond was not made to contribute to the family and community as much as his father. He doesn’t always need to work for money; he came from a communist country so he worked for community. Education was very important to him and becomes apparent through his child.He was a very strong, integrity, incorruptible, non-negotiable, religious and inflexible man. * 3. In ‘retrospect’ or hindsight Raimond is able to show his perspective contrasting to his fathers perspective. â€Å"Childhood as we know it† positions the reader to realize that childhood is a small time frame and many do not enjoy it such as his father. It makes people realize how hard times were back then. * 4. The use of past and present tense enables the author to show his perspective of things and relate things to present day situations. This also positions the reader to understand hindsight and retrospectively. 5. He was a working class opposed to middle class. This is because he is a blacksmith and works hard long hours to get money and gets by with minimalistic things. Romulus believes that she is snobbish and her activities such as going to the theatres were mistakenly snobby. Christine’s parents were able to accept him even though there was a distinct difference in class because of Romulus’ availability to clothes through the black-market. He was able to get food and clothes from the money he made. The money he earned allowed their relationship to be socially acceptable. * Chapter 2 * 6.The labels ‘immigrant’, ‘foreigner’ and ‘new Australian’ are used to set a tone of alienation. It creates a clear separation between the Australians and the â€Å"Bolts†. Romulus felt no anger or â€Å"no resentment or indignation, or any other response which depended on the assumption that he was owed something better. † He was able to reconc ile the terms above by having no negative feelings. He also held onto his â€Å"young dreams of a new life and so he saw his two years of bondage as a short interim,† * 7. (p. 13) Baringhup: hay day late 1800’s. Cheeses were famous won many prizes. Small town. 00 kids at school. Flourishing community and therefore belonging to community. 1950 became a ghost town. Rather sentimental, gaiter feels bad that the community has diminished. The juxtaposition between the two paragraphs of the heyday compared to the ghost town of 10 houses and convenience store. Camp swelled in gold rush. When they weren’t working, little to do. Movies on sat night and occasional dances hosted. Immigrants didn’t like the eucalyptus. * 8. The landscape became symbolic of Romulus’ sense of alienation because of the idea the Romulus feels like a stranger due to the environment.He doesn’t recognize the harsh and perceived blandness of the Australian scenery. He has a bur ning desire for the European atmosphere shown, â€Å"He longed for the generous and soft European foliage. † Romulus finds the gum trees, â€Å"Symbols of deprivation and barrenness† (p. 14) This shows him being alienated and feeling foreign due to the landscape and he feels that he would look â€Å"directly to the foliage and always turned away offended. † (p. 14) It is Romulus’ memories being based around the diverse and beautiful European environment that is one of the reasons he feels lost and separated in Australia. 9. Romulus establishes a sense of community in his new environment with the other Romanians, Hora and Mitru. â€Å"He asked the man who greeted any new arrivals whether there were any other Romanians. † In searching for them it shows his need to find a sense of belonging. â€Å"He sought them out and they became friends quickly,† depicts the idea that they all needed a companionship and somewhere to belong amongst the foreig n surroundings. This idea that it an essential part to the human spirit is demonstrated through the use of the concise phrasing and wording of this bonding.The lack of detail in this part shows that immediately they sorted out to find somewhere to belong and then after that was settled, other factors were now able to take place. This is reinforced by the principles of Maslow’s hierarchy of needs. It is through the searching for common people that Romulus shows his spiritual needs to belong and connect. * * Chapter 3 * 10. The landscape illustrates a feeling of belonging or alienation to a place. â€Å"A dead red gum stood only a hundred meters from the house and became for my mother a symbol of her desolation. This shows how the countryside invokes ideas of desolation and alienation into his mothers mind. The use of symbolism and irony of the Australian tree that is renowned for lasting through harsh conditions contrasting with Christine who has a mental breakdown and has to back away from many commitments and is viewed to have very little character and any sagaciousness towards going through a tough time. The peppercorns â€Å"were planted as though to mediate between local and European landscapes. † Shows that there is not only despondency with the outback but also a consciousness of belonging.These two quotes create a stark juxtaposition from the gum trees to the peppercorns, which reminded his father of home. This inkling is backed up with the description of Frogmore. The farm being a simple place with no electricity or running water, rats and snakes living under the house and 6-7 pounds for workers in the area creates the primitive existence. This contributes and reinforces the concept of isolation and desolation through surroundings. The use of the descriptive and almost poetic language emphasizes these points. Also Raimond retrospectively looks back at how his mother felt.These points demonstrate tensions between the desire to belong and alienation. * 11. Christine was always troubled and spoiled which lead to her suffering isolation. She stemmed to self-harm, then sort out reconciliation. She did this by looking for company or someone to belong to. She had an affair with Mitru, which didn’t last long. Due to her alienation, she felt deprived and overdosed on sleeping tablets. â€Å"She was alone, small, frail, walking with an uncertain gait and distracted air. † Shows how the separation has affected her. The use of visual imagery shows how small she is compared to the vast environment. †¦She appeared forsaken. † As though â€Å"†¦she had returned from the dead. † The use of Religious and biblical allusion create a mellow dramatic setting. People view her as a drama queen. Raimond writes the book in hindsight in a sorrowful manner. As though to give the reader the interpretation that he feels sorry for her and what she had to do. He tries to understand her feelings of isolation a nd seclusion. He tries to be sympathetic towards her in retrospect, even though at the time, she was widely considered pathetic. * 12. Romulus, as an immigrant from a European background, finds it hard to ‘fit in’ among Australians.This could be due to his character and strong moral codes, but also due to him just being different. Him being afraid of the snake, and â€Å"unused to the tinder-dry conditions† caused Romulus to set fire to a large proportion of the property. â€Å"My father was the focus of their hostile attention. † This goes to show that they do not accept him at this point in time and therefore does not belong to either the people or the environment. They all believe he is too different. This is juxtaposed with the change in shorter paragraph structure to emphasis the change in opinion.People â€Å"attributed his survival to my father’s prompt and sensible action. † This shows how he is not completely outcast and he does belo ng to certain parts of the society despite his differences and misunderstandings with some locals. * * Chapter 4 * 13. Hora’s connection with Romulus illustrates a deep sense of belonging and their combining for an enterprise. Hora however, does not respect Christine. There is a contrast between belonging with Hora and Romulus and his alienating attitude to Raimond’s mother. â€Å"He and Hora, together,† gives a hint of the bondage and belonging to one and other. They had quarreled and they quarreled,† deeply juxtaposes the idea of belonging and indicates a stark reality of separation and disconnection. â€Å"Heightened his impatience with my mother. † Shows that Hora has little respect for Christine. The constant argument between them also goes to show that he has such little respect for her. The emotive language used in â€Å"my mother was stricken with remorse,† helps to bring out the irony in the statement. The mother had not cared about her husband, was selfish, flirtatious with other men, exaggerated, had a false note and was incapable of sympathizing with another person yet she had â€Å"remorse†.We can see further disrespect through the use of prosaic and factual language: â€Å"Hora disliked her and did not respect her. † Also through the use of symbolism such as with the oranges, eggs and milk. They are symbols of looking after and a sense of responsibility, also the relationship tension between him and Christine. However a connection to Romulus and Raimond is shown through respect in contrast to disrespect with his mother. He goes to the effort to get oranges â€Å"each week he carried groceries, fruit and anything else we needed† p. 45 â€Å"my mother was unmoved by his efforts† p. 45.The eggs are a positive symbol of belonging, responsibilities from belonging, carrying the family, respect for Romulus and therefore belonging to family. It also is a symbol of livelihood. The milk s hows responsibility of wanting Raimond to eat right and again respecting Romulus’ ways. â€Å"Carried†¦in a heavy sack on his back† symbolizes him carrying the weight and responsibility of belonging to the family on his back. This is the upmost respect for Romulus, doing it whilst despising Christine in the process. Hora carrying them It is through these statements that we can clearly see the contrasting respect for Christine to Romulus. 14. ‘Pass auf mein lieber Fritz’ exemplifies the connection between Hora and Raimond. This represents Hora’s sense of responsibility and mark of respect to Romulus. â€Å"Our previously affectionate relationship,† shows a strong connection between the two. Although it changes slightly due to disciplinary action, he still is a ‘father figure’ and they find a feeling of belonging to each other. The use of a cliche â€Å"when I was sailing close to the wind,† emphasizes the belonging and respect that Raimond has for Hora, that he would never try to cross the border line.The relationship serves a purpose of showing belonging to each other but not belonging with place. Their close attachment through alienation, because they feel so left out, they form a close relationship with each other. The close bond between them is symbolic of him belonging to the family in replace of Romulus as a â€Å"step in father†. This close relationship is shown. * 15. Like most Europeans sets Romulus apart from Raimond because of the strong moral codes. The razor is important because it shows how strong Romulus’s moral code is. Raimond lied about steeling the razor.Romulus gets more ‘angry’ about him lying about it rather than steeling it as well. This shows the strength in his moral code and overreacts with a vicious beating of Raimond for a corruption in his character. We see the repetition or theme of character appearing in this part of the chapter. This focu s of character emphasizes the disconnection, difference and contrast between Romulus â€Å"like most Europeans† and Raimond. As Raimond didn’t see himself as European he feels the isolation and separation in pressure on strength of character.Romulus also feels disconnected as Raimond says retrospectively, â€Å"He feared that I would be like my mother. † This shows that Romulus feels Raimond is closer to his mother than him and could maybe belong better with her. The â€Å"cut throat razor† shows the difference in times as Romulus values the razor for its â€Å"craftsmanship† and aesthetic value. Raimond however, feels that it could be used to cut wood and doesn’t value it as much and therefore throws it in the river. This again shows the lack of belonging Raimond has to his fathers European background.Retrospectively he looks at them belonging to each other and he has utmost respect for his father even though at the time there is more tensi on and disconnection. â€Å"I knew my father valued truthfulness above most things,† This sets high moral codes which may have led to him not belonging due to the higher than normal standards and values set. Through this we can see Raimond and Romulus are separated through this strong moral code even though they may appear to belong. * 16. The relationship between father and son is strongly depicted through the Spartan description of Frogmore life.There is a change in language, the Spartan lifestyle shown by simple, banal and prosaic sort of language. This is a symbol of a beginning to belong to each other and possibly the environment. This contrasts with the next passage, which is crammed with rich and emotive language. This also shows a sense of connection and belonging to education and the belief in learning they posses. Romulus’ leg is mended and he is able to go back to his trade, being a blacksmith. This creates a sense of harmony. Romulus becomes connected, not with community but with his son, trade, past and background.Descriptive language highlights the new sense of belonging. â€Å"This was better than the poultry farming. †, â€Å"Classical hammer beaten iron work†, â€Å"Took such pride†, â€Å"crafted so superbly†, â€Å"copious amounts of fruit†. These use excessive adjectives, over description and exaggeration to create an appearance of harmony and peace to the reader. This helps the reader to become more attached to their relationship after times of adversity. This is a happy time for Romulus and therefore a happy time for his son. Present participles have an emotive effect all creating a sense of belonging between father and son.

Heredity, the Environment and Development Essay

The study of genetics has grown out of a desire to understand how exactly the individual comes to be just that, an individual different from its peers. In order to comprehend the scope of possible outcomes and how they came to be behavioral genetics looks at a number of variables; these include the impact of our genes (nature), and our environment (nurture). A countless number of hypotheses were put to the test through research to analyze the degree of influence of each. In this paper, team A will discuss the methods of behavioral genetics, the various research techniques used, their testing populations and why used, along with proposed answers and explanations. Behavioral GeneticsBehavioral geneticists uses family, twin and adoption studies as a basis for their argument of individual differences (Lerner, Bearer, Garcia, & Coll, 2004). A significant contributor to studies in behavioral genetics, provides this definition: â€Å"Behavioral genetics is the genetic study of behavior, which includes quantitative genetics (twin and adoption studies) as well as molecular genetics (DNA studies) of human and animal behavior broadly defined to include responses of the organism from responses measured in the brain such as functional neuro-imaging to self-report questionnaires† (Plomin,2004). Read more:  Influences that affect children’s development essay One of the first twin studies was conducted by Bouchard in 1979 when he found a set of monozygotic twins, babies from a fertilized egg that splits into two. The babies were separated at a few weeks old. The babies had many physiological and psychological similarities. Since Bouchard’s initial study it has been proven that, while monozygotic twins raised together have many similarities, those separated at an early age have an even greater likeness. Since twins being raised together are more likely to highlight their differences in order to maintain some element of independence, behavioral geneticists argue that this indicates a strong genetic underpinning in human development (Plomin, 2004). Research conducted by Grilo and Pogue-Geile (1991) correlated the familial relationships with extroversion. The study included monozygotic twins reared together and apart, dizygote twins reared together and apart, biological parents and children, biological siblings, adoptive parents and children and  unrelated siblings reared together. The results reflected that the highest correlation was between monozygotic twins raised together and apart. The lowest correlation was between unrelated siblings raised together. For behavioral geneticists, these results conclude that genetics are at work in determining the extroversion of a person. Behavioral geneticists suggest reasonable doubt in assuming connections between psychological environments and developmental results may be genetically arbitrated and that the environment a person is in responds to the genetically influenced characteristics (Plomin, 2004). The genetic association between parent and child is useful to examine. For example, â€Å"†¦differences in parenting can be the genetic effect rather than the environmental cause of children’s psychopathology† (Plomin, 2004, p. 345). The results of the twin, adoption and family studies support these assumptions. The Correlation of Heredity and EnvironmentThe nature versus nurture controversy exists because some people believe that a person’s genetics has the greatest impact on their personality, intelligence and behavior. On the flipside, some people believe that the environment has more of an impact. Behavior geneticists assume that behavior is influenced by the relations of heredity and environment. With the help of twin studies, and adoption studies researchers are working on understanding what molds a person into the individual he or she is today. Twin studies, using identical twins, are conducted to understand how biology influences traits and psychopathology in humans whose genotypes are the same (Haimowitz, n.d.).Twin studies also use fraternal twins who share half of the genes they acquire at conception which helps to compare the degrees of genetic influence such as intelligence and personality. Adoption studies take a look to see if adoptive children exhibit the behavioral and psychological traits of their adoptive parents, or those of their biological parents (Haimowitz). Any links to biological parents can be attributed to genetics, and any connection to adoptive parents can be  attributed to environment. Heredity-Environment correlations can be shown in three ways. One is the passive genotype-environment correlations. Passive genotype-environment correlation exists when a child’s biological parents are raising him or her (MacDonald, n.d.). An example of this situation could be Ana’s parents having the genetic predisposition to be intelligent and read skillfully leading one to believe that Ana will more than likely share these skills. Evocative Genotype-Environment Correlation occurs when a child’s genotype provokes a specific type of physical or social environment (MacDonald, n.d.). An example of this type of correlation: Andrew is artistic, and outgoing, he will elicit encouragement to try out for plays. Sheena is very athletic and competitive; she will be encouraged to go out for sports. Active genotype-Environment Correlations emerge when a child seeks out environments he or she will find compatible and stimulating (MacDonald, n.d.). An example could be that a child like Matilda, who has a gift of music, will seek a musical environment where she can expand on her talent. Scientist researching how genetics influences academic achievements show three ways heredity and environment could possibly be correlated. The three ways in which Meredith Phillips and a team of colleagues found genetics and environment to be correlated are passive correlation, active correlation, and reactive correlation. â€Å"Passive correlation: genes influence both a child’s environment and heredity (Phillips, Brooks-Gunn, Crane, Duncan, & Klebanov, n.d.,  ¶ 3).† â€Å"Active correlation: genes influence the environments that a child seeks out (Phillips et al.,  ¶ 3).† â€Å"Reactive correlation: environments react differently to people with different genetic profiles (Phillips et al.,  ¶ 3).† In passive correlation if a parent is the type of person whom enjoys reading, the love for reading could be transferred to the child from the parent reading to the child frequently. The child will already have the genetics from the parents. My son enjoys music I would like to think he received that from me because of my interest in music. He hears a great deal of music when at home therefore, the music rich environment my son is placed in has an influence on his musical achievements. Combined with the genetic aspect of my love for music and his father’s love for music the affect of his music rich environment causes a stronger influential desire to be involved in music. In active correlation, the child has genetic influences from the parent reading to him or her. When the child voices the desires for the parent to read to him or her, the parent enjoying the reading ultimately influences the child by reading to the child. The child requesting stories to be read is the incentive the parent has to continue the process along with the parent’s love for reading. Reactive correlation was described as genetics affecting the child’s physical features with the child’s features being judged by peers. The views of the child’s peers are voiced and in the process the child’s academic achievements are effected. The child’s environment can put him or her under a certain labels. In this situation genetics affects the views of the child’s peers and the environment combined with genetics can have a negative affect on the child’s academics. Definition of Shared and Non-shared Environmental ExperiencesBeyond genetics, each individual has a unique personality that is based on a blending of their shared and non-shared experiences in life. Shared environmental experiences are those which the majority of the world encounters. Shared experiences can occur differently by culture, but generally adhere to a specific â€Å"social clock or a set of age norms that defines a sequence of normal life experiences (Boyd & Bee, p. 10).† For American culture think of the traditions of school, watching a baseball game, having a BBQ, getting married, having children, working and retiring as relative shared norms that all, or most of us, encounter. Of course, non-shared experiences are different for each of us; these are categorized as individual experiences. Individual or non-shared experiences  can be influenced by â€Å"race, socioeconomic status, and other social factors (Boyd & Bee, 2006, p. 36).† These individual differences can also be related to school, relationships, marriage and childbirth and the unique perspective that each person has. Each individual thinks much differently and so the way that they perceive and interact in the world will make their experiences, shared and non-shared, unique to them. Role Played by Shared and Non-shared Environmental Experiences DevelopmentThe importance that shared and non-shared experiences have in development is that they help shape our personal development as well as our social development. If in fact, each person encounters shared experiences in accordance with the norm expected, they are more likely to fit in culturally and have a higher understanding of appropriate and healthy relationships. Likewise, with non-shared experiences if interactions each person has with their parents and peers, and in his or her independent life, is healthy they will know themselves internally and process environmental factors that occur around them in a healthy manner. If the shared and non shared experiences of an individual do not go according to the norms of society it will be more difficult for them to develop into healthy adults who function both independently and interdependently at appropriate levels. (Boyd & Bee, 2006, p. 36) In conclusion, behavioral geneticists have used a wide array of approaches to their research in developmental theories. Through the use of identical twins a great deal of information has been acquired on the basis of both genetics and environment. The strongest proponent of this argument was shown to be the case of identical twins separated at birth exhibiting very similar characteristics even though they had not been raised in the same environment. Corresponding research which also strengthens this argument shows that adoptive children exhibit very few of the characteristics of their adoptive parents. It seems safe to say that genetics lay the foundation of behavior with environment and individual experience capable of exhibiting some influence beyond that. References Boyd, D., & Bee, H. (2006). Lifespan Development. Retrieved from http://ecampus.phoenix.eduGrilo, C. M., & Pogrue-Geile, M. F. (1991). The Nature of Environmental Influences on Weight and Obesity: A Behavior Genetic  Analysis [White paper]. Retrieved from National Institute of Health: http://www.pubmedcentral.nih.govHaimowitz, A. G. (n.d.). Heredity versus Environment: Twin, Adoption, and Family. Retrieved April 21, 2009, from http://www.personalityresearch.org/papers/haimowitz.htmlLerner, R. M., Bearer, E. L., Garcia, , & Coll, C. G. (2004). Nature and Nurture: the Complex Intereplay if Genetic and Environmental Influences on Human Behavior and Development. . Retrieved from http://books.google.com/books?id=iFriCJCTsx4C&printsec=frontcoverMacDonald, K. (n.d.). PSYCHOLOGY 361: BEHAVIOR GENETICS. Retrieved April 21, 2009 , from http://www.csulb.edu/~kmacd/361Notes2.htmlPhillips, M., Brooks-Gunn, J., Crane, J., Duncan, G. J., & Klebanov, P. (n.d.). How Might Genetic Influences on Acad emic Achievement Masquerade as Environmental Influences?. Retrieved April 22, 2009, from http://www.children.smartlibrary.org/NewInterface/segment.cfm?segment=2606Plomin, R. (2004). Genetic and Developmental Psychology. Merrill-Palmer Quarterly, 50(3), 341-352. Retrieved from http://muse.jhu.edu/login?uri=/journals/merrill-palmer_quarterly/v050/50.3polmin.html