Analysis of SSR Sequences in Rice

Analysis of SSR Sequences in Rice 3. MATERIALS AND METHODS 3.1. Plant Material Total of twenty one introgression lines carrying African rice genes and its parent lines were used in this study (table 1) 3.2. Methods 3.2.1. DNA extraction Genomic DNA was extracted from young leaves of the seedlings grown in fields of Huazhong Agricutural University, Wuhan, China. Method used was according to Cetyltrimethyl ammonium bromide (CTAB) method reported by Xu et al. (2011), but with some improvements. Preparation of 1.5%CTAB buffer CTAB 3.0g 1M Tris pH8.0 15.0ml 0.5M EDTA (ethylenediaminetetraacetic acid), pH8.0 6.0ml NaCl 12.28g dd H2O 200.0ml PVP40Mw40, 000 1.0g Tris-base buffer to make sure pH=8 Two gram of fresh or frozen leaf tissue was ground to a fine powder in liquid nitrogen with a conical hand tissue grinder by using mortar and pestle. The powder was transferred to 1.5ml Eppendorf tube content 750ÃŽ ¼L of 1.5*CTAB and 25ÃŽ ¼L ÃŽ ² –mercaptoethanol. Then incubated for 1hour in a 65 ºC water bath with an interval mixing by inverting the tubes every 15min. Add 750ÃŽ ¼L of chloroform: Isoamyl alcohol (24:1) was done in a fume hood. The samples were gentle mixed by shaking for 30min in shaker at room temperature and then spin in the microfuge at 10000rpm for 10min to precipitate the cell debris. The upper aqueous phase (supernatant) was pipette and transferred into fresh eppendorf tubes. The same volume from preceding mix was put to the liquid in fresh eppendorf and the mixed shaking for 30 min in the shaker then spin in microfuge at 10000 rpm for 10 min. The supernatant was pipette and transferred to fresh eppendorf tubes, and then 2/3 the volume from iso-propyl alcohol 70% was added to each sample, then the samples was put in refrigerator 1 hour. After that centrifuged for 3 min with the rotational speed of 7000 rpm in 220c. After this step DNA collected on the tube sides. The supernatant was carefully discarded from the tube, DNA pellets appeared as tiny white tear drop-shaped smears on the tube sides. The pellets were washed twice with 75% ethanol, then air dried by inverting the tubes up side down over tissue paper. To re-dissolve DNA pellets 100ÃŽ ¼L of ddH2O was added to the tubes and immediately stored at –200C 3.2.2. Quality and quantity check of DNA DNA was checked for its purity and then quantified. The genomic DNA was run on 1.0% agarose gel stained with ethidium bromide and photographed under UV transilluminator using Image Lab TM software Version 4.0.1. The concentrations of the pure genomic DNA as assessed by agarose gel electrophoresis were estimated on spectrophotometer ND-2000. Based on the quantification data, all the genomic DNA samples were diluted to a final concentration of 100 ng†¢ÃŽ ¼L-1 with double-distilled water (ddH2O) and stored at -20o C for further use. 3.2.3. SSR analysis 3.2.3.1. PCR amplification and agarose gel electrophoresis A total of 50 microsatellite primer pairs were used for analyzing and identification 21 genotypes and then 22 polymorphic primers were selected to provide genetic identity and assess the genetic relationships among genotypes. PCR was performed in 20 ÃŽ ¼L reactions by using Thermal cycler touchdown as described by Don, et al. (1991), with some modifications. PCR mixture component The Touchdown-PCR program: PCR products were analyzed by electrophoresis using 2.0% agarose gel in 1x Tris Acetic cid EDTA (TAE), stained with ethidium bromide solution. then gel was visualized and photographed under UV light using Image Lab TM software Version 4.0.1. The SSR markers with high polymorphism were further used in SSR fingerprinting analysis 3.2.3.2. 6% denature polyacrylamide gel electrophoresis (PAGE) polyacrylamide gel electrophoresis has been used an unique analytical tool for many studies related to the identification of cultivars, species and F1 hybrids 3.2.3.2.1. Materials A. preparation of 6% PAGE for SSR analysis B. preparation of 10*TBE (Tris-borate EDTA) buffer Dissolved in 800 ml double distilled water, filtered through 0.22  µm filter paper, made up to 1000 ml. C. preparation of 40% Acrylamide d. Silver staining Preparation of silver solution 2.5g AgNO3 (Silver nitrate) 2000 ml Distilled water Then shaking well Preparation of developer solution 28g NaOH (Sodium hydroxide) 10ml HCHO (Formaldehyde) 2000 ml Distilled water 3.2.3.2.2. Method The large and short spacer glass plates, combs, and other pertinent materials were cleaned with water and completely dry. The inside of both plates were cleaned with 95% ethanol to facilitate drying. 650ÃŽ ¼L of Repel-silane was applied to long plate and spread evenly using tissue paper. 800ÃŽ ¼L of glide-acrylamide (200ÃŽ ¼L qin he gui wan + 10 ml 75% alcohol then agitate) was applied to short plate and distributed evenly using tissue paper and leaved to dry for a short time. Glass plates and sealers using clamps were assembled according to manufacturer’s instructions. 60ml of 6%PAGE, 40ÃŽ ¼L of TEMED (Tetra-methyl-ethylene-diamine), and 400ÃŽ ¼L of 10% ammonium persulfate (APS), previously stored at 40C were mixed into beaker. The gel was leaved to polymerize for about 1hr. After 1hr polymerization it was assembled in an electrophoresis unit. After cleaning the wall with 1X TBE buffer, the gel was pre-run at constant 1500w for 30 min to clean the gel and pre heat the buffer to about 60-650. 20ÃŽ ¼Lof PCR amplified product was mixed with 5ÃŽ ¼L of the loading buffer and added 5ÃŽ ¼L in each well. The electrophoresis unit was resumed and allowed to proceed at 1200w constant until loading buffer covered more than 3/4 of the distance. Finally the unit was disassembled and the gel was subjected to silver staining. Silver staining Gel was soaked for 20-30 min in staining solution with gentle shaking followed by washing in water for 3 sec. For developing color, gel was soaked in developer solution with gentle shaking until band appeared. The gel was rinsed in water for several minutes. Then the gel was air dried and visualized under normal light. 3.2.3.3. Data analysis 3.2.3.3.1. Analysis of polymorphism Unambiguous polymorphic bands were scored visually for the presence or absence of corresponding bands among the tested accessions. Stutter and background bands were excluded. Those SSR markers displaying no polymorphisms, non-specific banding patterns or without PCR products were discarded. Molecular data were prepared by scoring the SSR markers amplification profile as present or absent for each marker to generate a binary matrix. 4. RESUTS The study was designed to provide genetic identity for introgression lines carrying African rice genes by using molecular markers. Twenty one rice genotypes were used in this study (Table 1). A total of twenty two pairs of SSR primers distributed in 9 rice chromosomes were selected to analyze the twenty one genotypes. The sequence and the details of selected primers showed in table 2 4.1. Analysis of polymorphism SSR-PCR reaction system was optimized with 3% agarose gel electrophoresis and SSR markers were analysed with 6% denaturing polyacrylamide gel electrophoresis. Polymorphism was analysed with composited 22 pairs of SSR primers and optimized SSR-PCR system to determine the appropriate SSR markers applied in fingerprint mapping. Banding patterns generated by primer pairs RM310, RM213, RM202, and RM80 in 21 genotypes are shown in Fig 1.and Fig 3., while RM337 are shown in Fig 2. A total of 91 alleles were detected across 21 rice genotypes using 22 SSR markers. The maximum number of polymorphic alleles was 6, while the minimum number of polymorphic bands (2 alleles) was amplified with the markers RM 85, RM240. The average number of polymorphic alleles per marker was 4.1. Molecular data were ready for recording the SSR markers amplification profile as current (1) or absent (0) for each marker to create a binary matrix. The binary matrix data were analyzed through the use of the Similarity for Qualitative Data (SIMQUAL) module to generate Dice similarity coefficients [Dice LR. 1945]. The similarity coefficients were used to construct dendrograms using the Unweighted Pair Group Methods with Arithmetic means (UPGMA). 4.2. Cluster analysis The SSR markers were able to distinguish between different rice genotypes. The high degree of polymorphism of microsatellite markers allows rapid and efficient identification of rice genotypes. These markers classified the rice genotypes into eight clusters. (Fig4.) According to the results of pylogenetic tree twenty one genotypes were divided into eight groups according to the standard genetic similarity o.73. The lowest diversity was found between ILA65 and ILA78 (similarity level 98%) that strengthen the supposition of close relationship between them. While the highest diversity was found between J23B and other genotypes used in this study at similarity level 35%. These obtained results could be due to the number of SSR markers used in the study or the bias of genetic similarity estimation conducted by the UPGMA-based method. 4.3. Principal component analysis A principal component was performed using 22 SSR markers. The Values of the Eigen values and their contribution to variation are presented in Table 3. The score plot of 21 genotypes based on the first two principal components is presented in Fig 5. 4.4. Genetic similarity among Rice genotypes The Dic similarity was computed according to the obtained data from the polymorphic primers. The similarity index value obtained for each pair wise comparison among the 21 genotypes and presented in Table 4. The similarity coefficients ranged from 23.08% to 97.8% among tested genotypes. ILA 65 and ILA78 were the most similar among all the genotypes with a coefficient of 0.9780. The least similar genotypes were J23B and ILA19, ILA123 with a coefficient of 0.2308. 4.5. The fingerprints for identification The results showed the molecular identification of 21 Rice genotypes using 22 SSR polymorphic sequences. The thirteen Pair of these SSR primers were selected from the polymorphic primers which can amplify clear bands and have more alleles to identity nineteen introgression lines carrying African rice genes and three varieties. Table 5 The microsatellite assay generated cultivar-specific alleles in some of the genotypes screened; these used as DNA fingerprints for genotypes identification. This will be the assistance for the establishment and defense of proprietary rights and the determination of cultivar purity. The core SSR used to generate the fingerprint code of each used germplasm Table 6. 5. DISCUSSIONS In our study, microsatellite markers were used for investigating genetic diversity of 21 rice genotypes under study (Table 1). To this end, 22 primer pairs of microsatellite were used which had relatively high polymorphism in available literatures (Table 2). According to the previous results primer pairs will be referred to as loci and DNA bands as alleles (Sefc et al., 2000). The number of alleles obtained by microsatellite markers varied from 2 to 6 with an average of 4.1 alleles per locus. However, the average numbers of alleles detected in present study were significantly higher than this reported by JOSH et al. (2006) in non-Basmati aromatic rice genotypes of India which equals 2.6. The disparity among reports might be due to genotype number, SSR loci distribution, concerned sets of germplasm and gel electrophoresis method adopted in various studies. Higher number of alleles was found when a large number of landraces from a wide range of geographical origins were included in the study (Brondani et al., 2006). The cluster analysis, using unweighted pair group method of arithmetic means (UPGMA) was constructed for measuring genetic diversity and relatedness among the genotypes (Fig. 3). 5.1 cluster analysis based on SSR markers The similarity matrix was computed using SSR markers based on Dic’s coefficient following the UPGMA method using SHAN programme of NTSYS-pc. The Dic’s similarity coefficient for the SSR data set varied from 0.2308 to 0.9780. According to the results of phylogenetic treeà ¯Ã‚ ¼Ã‹â€ Figure1) Twenty one introgression lines and cultivars were divided into eight groups according to the standard genetic similarity which is 0.73. The first group ILA17, ILA13, STB, STA/F The second group ILA11 The third group ILA145, ILA21 The fourth group ILA12, ILA166, ILA1 The fifth group ILA147, ILB19, ILA172, ILA65, ILA78, ILA30, ILA29, ILA60 The sixth group ILA19 The seventh group ILA123 The eighth group J23B In group fifth we can distinguish two sub groups: sub group V-1 having ILA147, ILB19 and sub group V-2 having ILA172, ILA65, ILA78, ILA30, ILA29, and ILA60 but they were closely related groups. And five groups according to the standard genetic similarity which is 0.67. ILA17, ILA13, STB, STA/F ILA11, ILA145, ILA21, ILA12, ILA166, ILA1 ILA147, ILB19, ILA172, ILA65, ILA78, ILA30, ILA29, ILA60 ILA19, ILA123 J23B Rice genotypes clustered into eight well defined groups in accordance with their pedigree, probably due to the origin genetic of these materials, because most genotypes obtained from other genotypes and had similar pedigree. The studied Rice genotypes were showed existence of genetic diversity among 21 rice genotypes. The SSR markers played an important role in studying the germplasm diversity in rice (Yu et al., 2005). The results indicated that SSR analysis could be a better method to study the genetic diversity in rice. The highest genetic distance was found between ILA17 and J23B, where they held the first and last position of the dendrogram. On the other hand, the lowest genetic distance was found between ILA65 and ILA78 in the same group. This result consistent with the pedigree of these two ILs which shared a high proportion of ancestry (Table 1) SSR markers efficiently separated the rice genotypes into groups consistent with their origin and pedigree. Gerdes and Tracy (1994) explained that pedigree relationship can be used as an indicator to test the effectiveness of markers in determining relationships among breeding lines. Our results showed that the SSR markers were able to detect the extent of genetic diversity among rice genotypes used in this study. 5.2 Principal component analysis The principal component analysis study was also done using the subroutine EIGEN. The PCA results showed that the PC1 contributed 65.1389% followed by PC2 7.8560% and cumulative variance of first two PCA was 72.9948%. The results were close similarity of the results obtained based on unweighted pair group method with Arithmetic average (UPGMA) 5.3 similarity index A similarity matrix according to the proportion of shared SSR fragments was used to establish the level of relatedness between the tested genotypes. Pair-wise estimates of similarity ranged from 0.2308 to 0.9780 and the average similarity among all genotypes was 0.6807 (table 3.). Two genotypes ILA65 and ILA78 were the closest related genotypes with the highest similarity index of 97.8%. This was followed by 94.51% similarity between two pairs of genotypes ILA166 and ILA1. The lowest similarity (23.08%) was observed between genotypes ILA19 and J23B, ILA123 and J23B. As expected, J23B had the greatest dissimilarity with all the other tested genotypes. The similarity coefficients of J23B with all the other genotypes ranged from 0.2308 to 0.4396. It could be concluded that Simple Sequence Repeat markers could identify the different rice genotypes, and some of rice genotypes under investigation have probably originated from closely related ancestors and possess high degree of genetic sim ilarity. 5.4 DNA fingerprinting analysis Finally the thirteen pairs of SSR primers were selected from the polymorphic primers as the core set of SSRs (Table4) which could detect varying numbers of polymorphic bands. Their amplified bands were clear, legible, easy to count, and distinguishable from one another. These markers were distributed among 8 rice chromosomes. The microsatellites exhibited several bands that were shared among the check genotypes. Eight accessions (ILA123, ILA19, ILA21, ILA12, ILA145, ILA11, STA/F, STB) displayed unique bands in comparison with all other genotypes with different microsatellite markers. SSR markers analysis will help the identification and differentiation of introgression lines. The information will enable construct a DNA fingerprinting database of tested rice genotypes (Table 6). Construct a unique DNA fingerprints of the tested genotypes can distinguish each of the tested materials and provide basic guidelines for its conservation. CONCLUSION Through the present study, a total of 18 introgression lines carrying African rice genes and three Varieties were identified with specific SSR primer. DNA-based SSR markers revealed high genetic diversity among the genotypes and were able to differentiate them successfully. The similarity index values ranged from 0.2308 to 0.9780 Highest similarity (0.9780 ) observed between ILA65 and ILA78, whereas lowest similarity (0.2308) obtained between ILA19 and J23B, ILA123 and J23B. Thus, it can be inferred that more diversity was detected using SSR markers as it is evident from its similarity value. Results showed the high polymorphism and abundance of SSR sequences in rice. Total of 13 primers were selected to generate fingerprint of 21 genotypes Amany Kamel Elhabbak

Compaison of two poems †‘Night Over Birkenau’ and ‘Earrings’ Essay

This essay will be analysing and contrasting two poems relating to the Auschwitz concentration camp during WWII and how it diversely affected Jews. The first poem is ‘Night over Birkenau,’ a first hand experience poem written by Tadeusz Borowski to display the daily lives of Jewish prisoners in concentration camps and ‘Earrings,’ a second generation poem by Annette Bialik Harchik, written to inform readers how Jewish women were imprisoned whilst exploring aspects such as the racial discrimination by Nazi Germans in the Auschwitz concentration camp. By exploring these aspects it seems that the poems are written to give a voice to the deceased Jews who have died without having the chance to inform the world about the inhumane crimes inflict upon them deceitfully. From the start of ‘Night over Birkenau’ readers are given a clear insight to the poem’s setting, hence the title ‘Night over Birkenau.’ This gives readers the sense of insecurity as a setting in the night is likely to be more vile. This is then reinforced in, â€Å"Again the grim sky closes.† ‘Grim’ gives readers a sense of horror; and ‘sky closes’ evidently proves that the prisoners are captivated in this horror, it is inescapable. ‘Again’ reinforces that it is cyclic, constant and never ending within their hearts. The Jews’ feeling of horror is initially derived from the horrifying methods used by the Nazi Germans to exterminate them. These were methods such as the crematorium, â€Å"and the eyes of the crematorium blaze.† The word ‘blaze’ allows readers to interpret the intensity and aggressiveness of the flame but it is the use of personification in ‘eyes’ which emphasises the intensity of the heat as the eyes symbolise the shape of an igniting red and blue flame. The personification allows readers to interpret how slow the Nazi Germans have made the length of the termination process, as the eyes represent a quality of live humans. They were burnt alive which emphasises the agony that Jews had to go through. Just to die in the end. All this gives us a like comparison of hell. In my opinion, the Germans in the poem possess a character equivalent God’s, shown by their possessiveness over a wide nation, and, how they wiped them easily because of their ignorant, unjustified hate. There is further reference to the ‘eyes’ later in the poem which can effectively be linked to stanza four where Borowski refers to the eyes and the poison on the same line, â€Å"my eyes are poisoned from sleep.† This reinforces the idea that the prisoners were tortured in the methods of death used as poison takes a long time to distil in the body, slowly working its way through the body resulting in the victim left in agony. Alternatively, the reference to the ‘eyes’ made in the above quotes can mean that the Germans’ crimes were being watched and would not go without being known to the world (hence how we now know). The use of ‘eyes’ shows that they are being watched more widely than simply saying ‘eye’. Imagery of nature’s deceitful and unwilling characteristic is similarly portrayed in ‘Earrings.’ At the end of the poem, we are similarly shown how nature trying to contribute to the Germans’ crimes. This is shown on the ending stanza of the poem. Harchik does this by describing the holes dug in which the camp prisoners’ corpses to be buried in, â€Å"The empty holes,† and, on the next line they are described as ‘grown shut.’ This is a very effective way to use language, it emphasises nature’s attempt to stop the finding of the Jews’ corpses after the massacre. This is shown by the use of oxymoron to emphasise how the holes that the Germans have dug to mass bury the prisoners’ corpses is veiled by grass growing over the burial sites eventually levelling off with the rest of the earth. The idea of nature’s conceal of the Germans’ crimes is similarly portrayed in ‘Night over Birkenau.’ â€Å"Fog descends over Birkenau.† ‘Descends’ allows readers to see the slow approach of the ‘fog’ covering all signs of reality. The use of ‘fog’ allows readers to interpret the spreading of loss and confusion on its progression. The fog concealing Birkenau can symbolise the ignorance of the outside world. They were blinded by mere fog which the Germans used as a temporary veil to their crimes. The world’s ignorance is shown by the way the outside world did not have a basic outlook of what was actually happening inside this typical, yet deceitful looking camp. Day be day, the tension of: hunger;sickness and fear accumulated within the prisoners. Even their feet were not put to rest. But, instead, they had to wear wooden shoes. A source of pain and can sometimes prove fatal. And with all this people in the Auschwitz camp were losing sanity. They were afraid of dying. Not just dying; but not being found again. Borowski cleverly presents the loss of sanity to readers by the theme of astronomy, â€Å"blue Orion- lost among the stars.† For one, a setting in outer space is very effective as it reinforces the same theme as the night, as space, likewise, is pitch black. The real reason to why Borowski used this quote is because the Orion is visible to everyone, all around the world. Yet, historically, because of the business (and lack of care) the world gave no attention to the crimes which were happening. Physical loss is shown in ‘Earrings’ in stanza four. On the first line, Harchik shows readers how atrocious her mother’s physical condition is once she left her ‘earrings’ from the last stanza, â€Å"Under her wavy white hair.† The use of ‘white’ emphasises the dramatic change in physical attributes that happen within a character – this emphasises how much the camp prisoners must have been worked out. The use of the alliteration, ‘Wavy white,’ gives a double impact and meaning to the ageing idea, as similarly ‘wavy’ hair is often degraded in quality and is the last stage of hair ‘development’ within a person thus it is a characteristic of an ageing person. Fear is heavily referred to in ‘Night over Birkenau.’ It was the initial reason to why people in concentration camps lost sanity. This fear of death and humiliation had a huge psychological impact on the prisoners. Which, with the deprivation of food and water, depression and over working – it could only get worse. The first stanza describes the potential threat which the camp prisoners are facing, and this is done in very simple and deeply understood language, â€Å"Like a crouching beast over the camp.† The word ‘beast’ is used to portray a grim character which will do anything possible to harness its goal. This fear is further reinforced because a ‘beast’ also possesses a savage, daunting appearance. But fear is shown when Borowski uses the word ‘crouching,’ to show that prisoners lived under threat as the Nazi Germans have the potential to take you. Just like the beast. The use of enjambment helps to reinforce the beast’s potential tokill by the use of death imagery, â€Å"The moon sets pale as a corpse.† The colour of the moon is comparable to ‘corpse.’ The aftermath of death. Alternatively this can show fear by the way that prisoners could not look up to the sky for hope but misery. At the beginning of stanza two, Borowski shows readers the lack of defence prisoners had and their vulnerability to the Nazi Germans in, â€Å"And like a shield abandoned in battle.† The use of ‘shield;’ ‘abandoned’ and ‘abandoned’ in his simile allows readers to see the situation the Jews were in. They could not retaliate, as, without a shield, they could not defend themselves enough to make it a fair battle. They were betrayed; and their only chance of survival was by following their enemies’ instructions until their fates are decided. This quote can link with, â€Å"Lead foot crushing my chest.† ‘Lead’ is a metallic element, and without the ‘shield’ mentioned earlier, prisoners were not able to defend themselves against the metallic ‘crush’ attack style inflicted by the Nazis’ boots, making them resign for the humiliation. We can see this by the way the poet chooses to describe all images of fear and death figuratively to personify how they were mislead to the ‘gate of hell’, â€Å"The transports growl in darkness.† Borowski chooses to use ‘transports’ instead of simply saying ‘transport,’ to emphasise the abnormal amount of train activity in the camp. The fear is further reinforced by the use of onomatopoeia, ‘growl’ which can be linked to animal imagery of the beast in stanza one. The purposeful play of plurals is used similarly in stanza 4 to convey a different meaning, â€Å"Like God’s judgement on the corpse of the earth.† This only refers to one thing ‘corpse,’ instead of saying ‘corpses’ – this is done to show that the Jews’ unity. They all went through the same experience and they all die in the same place, they were all equal in such a tragedy. Fear is presented in a differently in ‘Earrings.’ Harchik identifies her family’s name on the first line; instead of keeping anonymous as Borowski does in his poem. This shows us that the intention of the poet was to speak out to give a voice to the Jewish women almost 45 years after the mass termination of the Jews in her poem published in 1989 with other similar poems speaking about Jewish women in the Auschwitz camp. The way in which the Jewish generation suddenly arises from its dormant state in decades shows how survivors were afflicted with fear from the experience they had to go through. ‘Earrings’ shows speaks out by, first of all, giving her family’s name, with the basis of the poem coming first hand from her mother’s perspective but is also based around other Jewish women, the general picture. In comparison to ‘Night over Brikenau,’ ‘Earrings’ surround itself around a more factual and authoritative theme thus it is less emotional. Readers are shown the poem’s factual content by Harchik’s straightforward language, which is effective and to the point, â€Å"stripped, shorn and tattooed.† Straightforward and to the point. Loss is presented in both poems. But, in ‘Earrings’ Harchik shows the readers the loss by deprivation of vital things to Jewish girls, such as culture. â€Å"Ears pierced in infancy; adorned in string golden hoops for girlhood; diamond studs for marriage.† Harchik shows the loss of identity and culture by the way that people, without her earrings, would not be able to differentiate whether she is married or not – they lost their identity. And the importance is shown by the clever use of semi-colons to emphasise how each step and every step was equal in importance and must be conducted in a woman’s lifetime and the deaths of Jewish children shows the deprivation that is caused by the Germans. Harchik explores the aspect of death in the concentration camp. This is shown in stanza three to show the death toll of the prisoner’s using the earrings as a symbol of their identity: â€Å"leaving behind her—earrings in a huge glittering pile of jewelry.† The word ‘huge’ emphasises the sheer amount of women imprisoned as they have all left their belongings in the pile. Harchik uses the word ‘leave’ to show the that her mother left her possessions with her intention – which reinforces the deceitfulness of the Nazi Germans as they lied and told the Jews that they were only going in for a ‘shower,’ when, in fact they were to be inhaled with toxic gas. ‘Glittering’ shows that the jewellery was new and polished – in other words, it was to give an impression. This can be referred historically to the way Jews were deceived into a wrong destination, the total opposite of what they have been told. A destination in which there minds would never dwell on. Alternatively, ‘glittering’ can suggest the speed at which everything is happening, which is very fast. Whilst the pile of jewellery is ‘huge,’ it is still ‘glittering’ to emphasise the activity of women leaving their possessions and never coming back out. Both poems portray the same sense of unity between the Jews making them ‘one’. This is shown in a simpler and more effective form in ‘Earrings’ because the link of unity between the Jewish women was also a link of culture as all the women carried at least earrings. Harchik shows us this by using the world ‘pile’ in, â€Å"in a huge glittering pile of jewelry.† ‘Pile’ suggests the disorganization in the camp – which reinforces the idea how everything in the camp was going at extreme speed. But, it is this ‘pile’ which suggests the equality that all the Jews had – every person’s possessions was mixed in a huge pile with other cheaper or more expensive items. There is a huge range of who’s this jewlerry may belong to whether it was a poor or a rich person. All that really mattered was that these people were Jewish. In ‘Night over Birkenau,’ Borowski displays the aspect of humiliation in a discreet way to emphasises his feelings. A clear example of this humiliation is shown in stanza three. Humiliation is shown here when the Nazi Germans stamped their shoes on the prisoners to show how their dominance over them as they are the ‘Arian race’ and how Jews were inferior in comparison to them. â€Å"This lead foot crushing my chest.†The use of words like ‘crushing’ is effective, it does not simply say ‘stamping’ but goes into a higher level of complexity to relate it to make it deadlier to emphasise that the Germans were never lenient with the camp prisoners. This can subtlety linked with the previous line of the Stanza ‘Breath rattles’, which is a sound of suffocation as the Germans crushed the prisoner’s chest. From this, we can deduce how Jews were racially ridiculed as soon as they entered the camp. They were not only humiliated but seized in a deadly way which is reinforced in the enjambment, â€Å"is the silence of three million dead.† This is what almost all Jews had to go through; regardless of who they were in the other world: the world were they had their own names. Prisoners humiliation is portrayed in ‘Earrings.’ Harchik guides readers through it in a chronological format where commas identify the pause between every step, â€Å"my mother was stripped,shorn,tattooed.† Harchik uses a continuous 3 line enjambment to show the speed at which everything is happening. Once prisoners are off the train they must do what the Nazi Germans tell them without a pause. The personal reference to her mother will make readers more sympathetic towards the situation since a mother is a character of significance. Prisoners were treated barbarically like a herd of sheep. We can see this by the way they had to follow all orders given to them, just like the sheep would do to the farmer without a halt. What is even more humiliating, is the way that the womens’ heads were shaven off completely, ‘shorn.’ This is done to remove any environmental variation between them. They could not be themselves – they were almost clones of each other serving the Germans. They were not easily differentiable and in some cases it was non-deferential between a man and a woman. And this mark of identity is shown by the way they were ‘tattooed’ – almost a sign from the Germans of their permanency in the camp. This shows the how some prisoners were even humiliated by their simple, indirect thoughts. There is also an effective use of alliteration in both ‘Night over Birkenau’ and ‘Earrings.’ Both of the poems use alliteration to convey both both the onomatopoeic meaning and a hidden historical inference. In ‘Night over Birkenau,’ alliteration is used to show death â€Å"It’s steamy stifling.† This refers to the sound of a snake ‘Ss’, which is an example of animal imagery with the intention to bring death. The use of ‘steam’ emphasises the intensity of temperature whilst ‘stifling’ reinforces the idea of the Germans’ control. Despite the ‘living’ conditions they are in – they are forced to live through it. The punchy full-stop is used to show an end to the life of prisoners who chose to give up than live under control. Alliteration is similarly used in ‘Earrings’ to show the camp’s treatment towards prisoners â€Å"My mother was stripped,shorn.† The alliterate use of ‘Ss’ is used here again, ‘striped,shorn.’ The main use of alliteration here is because of its historical reference. ‘SS’ stood for ‘Schutzstaffel’, a major Nazi military organisation run by Adolph Hitler during World War 2 – this organization was primarily responsible for the crimes for the crimes against humanity, thus was the major inflict of pain and suffering to the Jews. Onomatopoeia is used in ‘shorn’ and when read gives a ‘Shhh’, a sound of sheering – which reinforces that the women prisoners’ heads were shaved off. Reference to religion in poetry helped prisoners to keep sane during very demanding times. With the prisoners’ loss of identity, culture and even their name there would only be one thing in which they can truly embrace in their hearts without the feeling of a threat, which is religion. In ‘Night over Birkenau’ religious issues are raised and introduced in the first stanza to set an instant scene of terror, â€Å"Grim sky closes circling like a vulture over the dead silence.† The idea of the sky ‘circling’ in vulture like motions, in Jewish scriptures is a sign to determine the Day of Judgement. They felt that this was the end of the Jewish spurt. And indeed; it was the ‘Day of Judgement’ for them – this was the Nazi Germans’ intention. To wipe out all Jews from existence. Alternatively, the quote can be used to show the death by animal imagery. The Nazi Germans are seen as the ‘vulture’, circling around its prey, the Jews. This simile further emphasises the vulnerability of the prisoners. Borowski chooses a vulture to show how strongly patrolled the prisoners were. Whilst they may think that they are safe – they are still watched cautiously by the Nazi Germans. On stanza three, readers are evidently shown the death. Not figuratively but statistically. â€Å"Is the silence of three million dead.† Furthermore, these quotes can link us as to how Borowski consciously gives evidence to the silence in the camp emphasising the Jews fearfulness speak. Silence is shown when Borowski coherently uses death imagery and death implies silence – this is evident by the poet in the fourth stanza, â€Å"like a vulture over the dead silence;† and this is reinforced by â€Å"Is the silence of three million dead.† In comparison with ‘Earrings,’ there is no involvement of religion.’Earrings’ concentrates on culture and identity. ‘Night over Birkenau’ is structured chronologically. The first stanza gives readers an insight to the poem’s setting including the thematic information to give the reader an idea as to what the poem is about. The first two lines of the second stanza progress to tell the reader about the emotions that the people felt; including their loss which is an effective follow-up from stanza one’s description of the night, the last two lines of stanza two show us the feelings of the people on the way to enter the camp, ‘The transports growl in darkness.’ Stanza three concentrates on the suffering and humiliation that the camp prisoners went through. The image of death is effectively shown throughout this stanza and there is an effective use of punctuation to reinforce death and tension, ‘It’s steamy, stifling. Sleep is a stone.’ Contracted form is used to reinforce the idea of ‘stifling’ as it can be used to show the suffocation and lack of space the prisoners had. The full-stop at the end of ‘steamy, stifling.’ can be used to show an end of life to some prisoners. The obvious idea of death is shown at the end, ‘is the silence of three million dead.’ Stanza four is cyclic to stanza one and gives the repetition of night, ‘Night, night without end. No dawn comes.’ The repetition of the night shows us the routine of the prisoners’ daily life in the camp but an alternative interpretation would be that the prisoners did not have hope, and hence ‘No dawn comes.’ Dawn is the opposite of night, thus it must mean hope and the dawn not coming reinforced the idea that hope did not come. Also, notice how there is a consistency in the number of lines in each stanza – this can also show how there was the same daily routine in the camp, Borowski makes everything in the poem repetitious just to show the number of times these crimes have happened without the world knowing – the prisoners did not have a voice. We can see Borowski’s cyclic approach from the beginning of the poem, â€Å"Night again. Again the grim sky closes.† The use of repetition, ‘again,’ reinforces the idea of routine and the tediousness of camp life. This is also effective when the poem is actually being read as ‘again’ is repeated twice. This is similarly portrayed in the ‘Earrings.’ Both poems follow the same gradual chronology – birth to death. This is slightly different in ‘Earrings.’ Harchik does not set a scene but ironically shows us the earrings as being a part of the prisoners’ life, it was a sign of hope for them. Harchik instead uses ‘tabbing’ structure to separate the ‘earrings’ away from the poem to similarly show the lack of hope but it is now shown ironically in comparison to ‘Night over Birkenau’ as the object of hope is now away rather than the object of misery being within the text. This can be interpreted to show how far away it was for the prisoners to be heard by the rest of the world and the lack of voice they received. ‘Earrings’ is a personal second generation poem and therefore the language nor the structure did not touch on sensitive issues that the Jewish generation in general felt, but bases it on what Annette Harchik’s mother. The ‘tabbing’ structure is heavily relied on by the poet to describe what it was before and after the entrance in the camp. The first stanza talks about her own family’s perspective and we notice how it consists of 3 lines to show that it has a lower priority in comparison with stanza 2 which gives background information relating to all the women in the camp and thus given four lines. The third stanza is the biggest with the factual content of what has happened to her mother, ‘my mother was stripped.’ Although Harchik bases this on the mother, she does go forward to give the idea on the wider perspective of what has happened to the people on the last line of stanza three, ‘in a huge glittering pile of jewelry.’ The enjambment used from the last line follow on from ‘earrings’, the symbol of the people – this is effective use of the enjambment because it links very well to the ‘huge glittering pile,’ which again evidently shows that a wide number of people were involved. Punctuation in ‘Earrings’ is less intense than ‘Night over Birkenau’. Harchik uses punctuation to separate different thing in the camp in a ‘step to step’ format, â€Å"mother was stripped, shorn, and tattooed.† Notice how there is a comma between every step that her mother has to go through, this is effective to the reader as they have to ‘pause’ between every step. This can also be interpreted to show the sheer humiliation that the camp prisoners had to go through – the pause emphasises that the prisoners had to stand there waiting for the next step to be conducted right in-front of crowds upon crowds of other, humiliated people. Lack of hope is similar between both ‘Earrings’ and ‘Night over Birkenau,’ both poems repeat the ‘symbol’ which is bringing misery or ironically in ‘Earrings,’ hope. In ‘Earrings,’ we witness the presence of the word ‘earrings’ in the first stanza to start the poem and the third stanza where the conflict is being described, once again the ‘tabbing’ structure between the text and ‘earrings’ emphasises that there is a lack of hope; which is similarly show in ‘Night over Birkenau’ but with the text to show the lingering presence of evil.. From studying these poems. I can evidently see that Borowski is a poet who has went through a lot of pain and agony before having the courage to show the world exactly what happened to Jewish prisoners during WW2. This is shown in his poem by the coherent references to methods of deaths, torture and disturbing images. We can learn from the poem, and his character is that living through demanding times – like a concentration camps can totally change one’s state of mind – and this is why Borowski has a coherent reference to death imagery – to feel like he’s not the only one. But even with all this, it was amazingly hard to escape the camp, but even harder to escape the mental scars which the camp’s life has subsided within him; resulting in an arousal of his suicidal feelings, and hence this mental disturbance caused him to commit suicide at the mere age of 28 despite being freed from the camp. Both poems allow us to relate to the massacres that have occurred during World War II to the Jews, as both flow progressively in a structured order. ‘Earrings’ has successfully explored the identity, culture and humiliation aspect in stanza three, whilst on the other hand, ‘Night over Birkenau’ heavily related to the fear and the death. In my opinion, the poet is trying to show readers that nothing exceeds humiliation, loss of culture and identity. And hence why Harchik after many years after the Holocaust she merely goes into the aspect of death but instead comments on the humiliation, loss of identity and culture. Her determination to inform the world is evident in her poetry. Her determination to tell the world about the Germans’ crimes gained her credaince to publish ‘Earrings’ in â€Å"Blood to remember† a 21st century version of the original ‘Earrings’ with a wider use of structure to further convey the loss of people. Personally, I preferred studying ‘Night over Birkenau’ as Borowski conveys his meaning to readers in a very vivid way. It is very inspiring and makes me think about how horrible it must have been to be in such a demanding situation. ‘Night over Birkenau’ allowed me to see how the world can be very deceitful to obtain and help who it wants and the constant repetition to the silence allowed me to think about how can the outside world not pay any attention to anyone. Not even a threatened nation fore coming the plague of World War 2’s plagued war-zone.

Alcoholics Anonymous Meeting Observations Of A Interview

Alcoholics Anonymous Meeting Observations Per my customary tradition of attending anything for the first time I arrived early to survey the environment as well as obtain a seat that would provide the most optimum observation vantage point. The alcoholics anonymous (AA) meeting I attended was held at the House of Disciples Life Recovery Center, a ministry of Wiseman Ministries. Interestingly enough, it used to be a funeral home. As I walked through the doors the first thing I noticed was a faint but distinct smell of burnt coffee, tobacco, and donuts. Making my way deeper into the meeting hall I located the source of the smell. After helping myself to a cup of java I walked over to a table sitting in front of chairs placed in a†¦show more content†¦The leader read from the AA book and afterwards invited anyone to share what was on their mind. After what seemed a lifetime, an elderly man raised his hand and then, after a nod from the leader, he stood up saying, â€Å"Hi. I’m â€Å"Bob† and I†™m an alcoholic†. â€Å"Hi Bob†, the group said together. Bob then began sharing a lifelong struggle with alcohol that started in high school as a teenager and spanned a course of some twenty years. Two decades of conflict and disharmony that seemed to connect with many of the attendees displayed by their numerous head nods and looks of shared shame. He said he never intended to become an alcoholic. He was just trying to â€Å"fit in† with friends. Another reason given for using alcohol was to squash the stress and anxieties all too familiar with the formative years of pre-adulthood. â€Å"Little did I know†, he said that he was setting the stage for how to handle all his subsequent problems later in life. Problems with his parents, he would drink. Disagreement with his girlfriend, he would drink. Bad day at work, he would drink. No matter what difficulty he encountered or strain life would throw at him alcohol was always the answer. This, on top of the good times. 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Transparency in Apple’s Corporate Statements - 3574 Words

Apple, Inc.: Transparency in Corporate Statements About the CEO On June 9, 2008, at the Worldwide Developers Conference (WWDC) in San Francisco, numerous media outlets and a packed house waited to hear Apple Chief Executive Steve Jobs’s keynote address introducing the new iPhone 3G. While the product’s debut drew raves worldwide, the real buzz was created by how gaunt and exhausted Steve Jobs appeared on stage. Rumors immediately began circulating on Internet forums on the apparent source of his condition – perhaps a reemergence of the pancreatic cancer he suffered in 2004 or some new unknown ailment plaguing the widely revered business icon? Six months after the WWDC, Apple announced that Jobs would not be delivering his customary†¦show more content†¦Since Jobs’s gaunt appearance at the WWDC in June 2008, Wall Street has been keeping a close eye on Apple. Its share price has been shifting with the different emerging announcements. While there is a segment of the public that feels that a person’s medic al condition is a matter of privacy and not under the realm of public disclosure, there is a segment that insists that, in this case, it is indeed within the purview of shareholders. Michael Hiltzik of the Los Angeles Times emphatically stated that â€Å"Apple has allowed its institutional arrogance, its culture of secretiveness, and possibly its solicitude for and fear of Jobs to lead it down a path of rank corporate irresponsibility.†4 Apple is facing credibility issues with both its corporate communication and with concerns regarding the company’s success without Steve Jobs. As Dylan Ratigan stated on CNBC’s Fast Money, â€Å"†¦the quality of information we’re getting from Apple is inconsistent at best and misleading at worst.†5 Combined with post-Jobs jitters, it seems unlikely that such a highly praised, successful company with an extremely loyal customer base would invoke such backlash and concern about its future over any single employe e, but clearly Steve Jobs proves to be the exception. Early Days Apple Computer was established on April 1, 1976, by Steve Jobs, Steve Wozniak and Ronald