Protective Role of Isolated Rat Hepatocytes Against CCL4

Protective Role of Isolated Rat Hepatocytes Against CCL4 DISCUSSION CCl4, Paracetamol and Anti-tubercular drugs cause ROS mediated cellular damage especially in liver, the site of metabolism of these drugs. During the regular physiological functioning the cells/tissues/organs use oxygen and various nutrients to generate energy. The free radicals are also generated in this process as the reaction intermediates. These free radicals may be very useful because they may promote beneficial oxidative processes. However the higher quantities of such radicals like superoxide anion (O2à ¯Ã¢â‚¬Å¡Ã‚ ·), NOà ¯Ã¢â‚¬Å¡Ã‚ · radical, and hydroxyl ion radical (à ¯Ã¢â‚¬Å¡Ã‚ ·OH), NOOà ¯Ã¢â‚¬Å¡Ã‚ ·, etc. may interact with the membrane lipids leading to lipid peroxidation and attack the DNA resulting DNA strand breaks. The lipid peroxidation also damage cell membrane resulting in the leakage of enzymes into the blood stream. Therefore the elevated biochemical levels are treated as biochemical markers of tissue damage. In addition the extent of lipid peroxidation is d irectly proportional to the tissue damage [Devasagayam et al., 2003]. There are certain inbuilt protective mechanisms, tissue enzymes GSH, SOD, CAT etc. which are involved in the process of combating free radical induced tissue damage. Over powering the inbuilt protective mechanism due to excessive generation of free radicals may lead to destruction of the tissues/organs [Tiwari, 2001]. Antioxidants are the chemical constituents, which are used for inhibiting the tissue damage by countering the free radicals; most of the antioxidants available in the markets are from natural origin e.g. vit-E, vit-C, tocopherol, quercetine, à ¯Ã‚ Ã‚ ¢-carotene etc. In addition there are reports that polyphenolic compounds like flavonoids, tannins are useful as antioxidants and organ protectants. Therefore many researchers are attempting to screen the herbs and herbal preparations containing polyphenolic compounds for organ protective properties. Mruthunjaya, 2008 in his study reported the presence of very high amount of polyphenols and gallic acid, a well known potent antioxidant [Gow- chin et al, 2002] as an important constituent of the roots of a widely grown plant L. inermis. He also reported the antioxidant and in vitro and in vivo hepatoprotective activity of L. inermis root extract against CCl4 induced hepatotoxicity in rats. So in the present study L.inermis was selected f or investigating the in vivo and in vitro hepatoprotective activity against drugs induced hepatotoxicity. Before screening the test extract for in vivo hepatoprotective activity the extract was subjected to the acute toxicity studies as per OECD guidelines 425 (limit test). The LIALC was found safe up to 2000 mg/kg as evident by the absence of mortality in the treated group. Hence, 1/10th (200 mg/kg) and 1/7th (300 mg/kg) of the safe dose were selected for the in vivo study. In vitro hepatoprotective Screening In the present study, isolated rat hepatocytes were used. The objective of this study was to confirm the protective role of test extracts against CCl4 induced hepatocytes damage. The CCl4 metabolized in (liver) endoplasmic reticulum and mitochondria with the formation of CCl3O (unstable complex radical), by CYT P-450. The nascent oxygen O resulted via lipoperoxidation causes rise in intracellular reactive Fe2+ ions, aldehyde and depletion of GSH, and calcium sequestration. Oxidative CCl3O, also by direct covalent interaction induces degeneration in Ca2+ sequestration. Failure into sequestration result in increased intercellular Ca2+, aggregation by proteolytic enzymes and cause an increase in Fe2+ ions, which in turn by lipid peroxidation precipitates aldehyde cytotoxicity [Zimmerman Hayman, 1976, Agarwal et. al., 1983]. Due to this membrane integrity is lost ultimately leading to necrosis. The loss of membrane integrity can be measured by Trypan blue exclusion assay in which the viable cells exclude the dye whereas dead cell take up the dye owing to the alteration in membrane permeability. Due to loss of membrane integrity cytosolic enzymes are leaked into media and the measurement of these cytoplasmic enzymes in the media serves as ideal markers for evaluating the extent of hepatocyte injury and protection offered by extracts. From the results it was apparent that both LIALC and LIAQ protect the hepatocytes against CCl4 damage as evident by the reversal of cell death and inhibition leakage of cytosolic enzymes. As discussed earlier this strong hepatoprotective activity of LIALC might be due presence of polyphenols and gallic acid which are potent free radical scavenging agents and antioxidants [Gow- chin et al, 2002] In vivo hepatoprotective activity Paracetamol (N-acetyl-p-aminophenol) is a widely used analgesic and antipyretic drug and is safe when used in therapeutic doses. However over dosage of Paracetamol is known to be hepatotoxic and nephrotoxic in man and in experimental animals [Parmar et al., 1995]. Paracetamol is a direct hepatotoxin i.e. intoxication is dose dependent and reproducible [Tygstrup et al., 1996]. Exposure of animals to higher doses produces centrilobular or massive hepatic necrosis followed by congestion and failure. The hepatic necrosis is associated with damage to sub cellular organelle including mitochondria. Thus the drug is used as a typical hepatotoxin to produce hepatic failure experimentally [Sing et al., 1999]. At lower doses, about 80% of ingested Paracetamol is eliminated mainly as sulfate and glucoronide conjugates before oxidation and only 5% is oxidized by hepatic CYT P450(CYP2E1) to a highly reactive and toxic electrophile i.e. N-acetyl-p-benzoquineimine (NAPQI). After over dosage of Paracetamol the glucoronidation and sulfation routes become saturated and as a consequence, Paracetamol is increasingly metabolized into NAPQI [Remirez et al., 1995]. Semiquinone radical, one-electron reduction metabolite of NAPQI mediates the cytotoxic effects of NAPQI. Production of these toxic semiquinone radicals is catalyzed by the microsomal CYT P450 reductase. These semiquinone radicals, in turn, can bind directly with cellular macromolecules to produce toxicity or alternatively, the radical can be reoxidized back to their original quinones by donating one electron to molecular oxygen under aerobic conditions. This donation of one electron then generates reduced oxygen radical species and hydroxyl radical. Both semiquinone and oxygen radical are known to be responsible for cytotoxic effects observed with quinones. Also NAPQI is detoxified by glutathione (GSH) to form 3-(GSH-S-yl) acetaminophen. Paracetamol overdose saturates the nontoxic metabolic pathway, i.e. sulfation, glucuronidation, and detoxification of NAPQI by glutathione. The reactive NAPQI may oxidize and arylate cysteinyl thiol group, forming adducts which inhibit the function of cellular proteins. Adducts formation has been demonstrated for a selenium-binding protein, for microsomal subunit of glutamine. Other mechanism, such as oxidation of pyridine nucleotides and lipid peroxidation, may contribute to cell damage by Paracetamol overdose [Tygstrup et al., 1996]. Nevertheless at high doses of Paracetamol, NAPQI can alkylate and oxidize intracellular GSH and protein thiol group, which result in the liver GSH pool depletion and the reactive intermediate reacts with other nucleophilic centers of vital molecules in liver cells leading subsequently to hepatotoxicity. In addition Paracetamol is also shown to directly inhibit cellular proliferation, induce oxidative stress, resulting in lipid peroxidation, deplete ATP levels and alter Ca++ homeostasis; all of these changes are considered potentially fatal to the cell [Sing,. 1999 and Remirez et al., 1995]. Anti-tuberculosis drugs act as inducers of hepatic CYT P450 enzymes. For example, Rifampicin is a potent inducer of CYP2D6 and CYP3A4, and Isoniazid induces CYP2E1 [Trevor, 2004 and Vuilleumier et al., 2006]. The induction of CYT P450 enzymes is known to take part in increased drug disposition and development of multi-drug resistance. Xenobiotics, including anti-tubercular drugs, undergo biotransformation in the liver catalyzed by microsomal enzyme systems [Bradford, 2005; Dai et al., 1995and Jeong, 1995]. Because of above reasons, Paracetamol and anti-tubercular drugs cause ROS mediated hepatotoxicity and elevate serum enzymes levels like SGOT, SGPT, ALP, LDH and Bilirubin content in animals following administration of anti-tubercular drugs or toxic dose of Paracetamol. Also these drugs cause depletion of endogenous antioxidant enzymes like GSH, SOD, CAT etc and abnormal levels of lipid peroxidation products.The results obtained in the present are in accordance with understanding as discussed above. i.e the serum enzymes levels like SGOT, SGPT, ALP, LDH and Bilirubin contents were significantly increased, GSH, SOD, CAT levels in liver depleted and abnormal levels of lipid peroxidation was observed in Paracetamol and anti-tubercular drugs treated animals. Estimation of serum enzymes SGPT is a cytosolic enzyme primarily present in the liver. The level of SGPT in serum increases due to leakage of this cellular enzyme into plasma by hepatic injury [Chenoweth Hake, 1962]. Serum levels of SGPT can increase due to damage of the tissues producing acute hepatic necrosis, such as viral hepatitis and acute cholestasis. Alcoholic liver damage and cirrhosis also can associate with mild to moderate elevation of transaminase [Harsh Mohan, 2002]. SGOT is a mitochondrial enzyme released from heart, liver, skeletal muscle and kidney. Liver toxicity elevated the SGOT level in serum due to the damage to the tissues producing acute necrosis, such as severe viral hepatitis acute cholestasis. Alcoholic liver damage and cirrhosis can also associate with mild to moderate elevation of transaminases [Harsh Mohan, 2002]. In case of toxic liver, ALP levels are very high, which may be due to defective hepatic excretion or by increased production of ALP by hepatic parenchymal or duct cells [Harsh Mohan, 2002]. LIALC reversed these enzyme levels indicating stabilization of cell membrane by preventing the damage due to free radicals generated by Paracetamol and anti-tubercular drugs. Estimation of bilirubin: In case of toxic liver, bilirubin levels are elevated. Hyperbilirubinemia can result from impaired hepatic uptake of unconjugated bilirubin. Such a situation can occur in generalized liver cell injury. Certain drugs (e.g., Rifampin and Probenecid) interfere with the net uptake of bilirubin by the liver cell and may produce a mild unconjugated hyperbilirubinemia [Rubin, 1995]. Bilirubin level rises in diseases of hepatocytes, obstruction to biliary excretion into duodenum, in haemolysis and defects of hepatic uptake and conjugation of bilirubin pigment such as in Gilbert’s disease [Harsh mohan, 2002]. Significant reversal of elevated bilirubin level in Paracetamol and anti-tubercular drug treated animals by LIALC indicated the strong hepatoprotective activity of LIALC. Estimation of Protein A reduction in total serum protein observed in the Paracetamol and anti-tubercular treated control rats may be associated with the decrease in the number of hepatocytes which in turn might result in decreased hepatic capacity to synthesize protein. But, when the LIALC was administered along with Paracetamol or anti-tubercular drugs a significant increase in protein content was observed indicating the hepatoprotection of LIALC. Estimation of endogenous antioxidant enzymes and Lipid Peroxidation Cells have a number of mechanisms to protect themselves from the toxic effects of the ROS. SOD removes superoxide (O2) by converting it to H2O2, which can be rapidly converted to water by CAT and Glutathione peroxidase. In addition, a large reserve of reduced glutathione is present in hepatocytes and red blood cells for detoxification of xenobiotics or free radicals. However, oxidative stress results in toxicity when the rate of which the ROS are generated exceeds the cell capacity for their removal. Lipid peroxidation is an autocatalytic process, which is a common consequence of cell death. This process may cause peroxidative tissue damage in inflammation, cancer and toxicity of xenobiotics and aging. MDA is one of the end products in the lipid peroxidation process. In order to elucidate the protection mechanism of LIALC in Paracetamol and anti- tubercular Drugs induced rat liver, lipid peroxide levels and anti-oxidative enzymes activities were analyzed. GSH is widely distributed in cells. GSH is an intracellular reductant and plays a major role in catalysis, metabolism and transport. It protects cells against free radicals, peroxides and other toxic compounds. GSH is a naturally occurring substance that is abundant in many living creatures. It is well known that a deficiency of GSH within living organisms can lead to tissue disorders and injury. For example, liver injury included by consuming alcohol or by taking drugs like Paracetamol, lung injury by smoking and muscle injury by intense physical activity, all are known to be correlated with low tissue level of GSH. The SOD converts super-oxide radicals (O2_) into H2O2 plus O2, thus participating in the enzymatic defense against oxygen toxicity. In this study, SOD plays an important role in the elimination of ROS derived from the peroxidative process of xenobiotics in liver tissues. CAT is the key component of the anti-oxidant defense system .Inhibition of this protective mechanisms result in enhanced sensitivity to free radical induced cellular damage. Administration of LIALC increased the activity of catalase in Paracetamol and anti-tubercular drug induced liver damage to prevent the accumulation of excessive free radicals and thus exhibited protection against Paracetamol and anti-tubercular drug. The observed increase of SOD, CAT and GSH activity in liver suggests that the LIALC extract have an efficient protective mechanism in response to ROS. And also, these findings indicate that LIALC may be associated with decreased oxidative stress and free radical mediated tissue damage. In our study, elevation in the levels of end products of lipid peroxidation in liver of rat treated with Paracetamol and anti- tubercular drugs were observed. The increase in MDA level in liver suggests enhanced lipid peroxidation leading to tissue damage and failure of anti-oxidant defense mechanism to prevent formation of excessive free radicals. Treatment with LIALC significantly reversed these changes. Hence it may be possible that the mechanism of hepatoprotection of LIALC is due to its antioxidant effect. Dept of pharmacology, P.R.R.M college of pharmacy,kadapaPage 1

The poem Two Scavengers in a Truck, Two Beautiful People in a Mercedes

The poem Two Scavengers in a Truck, Two Beautiful People in a Mercedes is a descriptive poem by Lawrence Ferlinghetti. Night of the Scorpion & Two Scavengers in a Truck, Two Beautiful People in a Mercedes The poem Two Scavengers in a Truck, Two Beautiful People in a Mercedes is a descriptive poem where the poet (Lawrence Ferlinghetti) observes two garbage men in San Francisco and two well-paid people in a Mercedes. The poet's observation is really about the way the garbage men look at the well-paid people and the way the rich people look at the garbage men when waiting at the traffic lights. This poem is about two garbage men and two posh office people in the Mercedes who stop at the same traffic light at the same time on the same day. The main reason I enjoyed this poem was because of the description of each person in the poem. One description I really liked was the description of the two people in the Mercedes, '' The man in the hip three-piece linen suit with shoulder-length blond hair & sunglasses The young blond women so casually coifed with a short skirt...

How did Shakespeare appeal to his audience, both in the 17th century, and in the modern era? Essay

Shakespeare’s play, Macbeth, created a big impact when it was written and first performed and it is still popular and well known. So why do so many people still enjoy it? What did Shakespeare do to capture his audience so well? Act 1, Scene 3 is the scene in which the witches meet again. One tells a detailed story about a sailor whom she is planning to torture. This shows the audience how mean the witches can be so the audience isn’t true how trustworthy they are. Soon after, the witches meet with Banquo and Macbeth and they tell them both prophecies; calling Macbeth Thane of Glamis (which he already is), Thane of Cawdor, and they say that he â€Å"shalt be king hereafter† (line 58). At the end of the scene, Ross and Angus come to inform Macbeth that he is thane of Cawdor. It is once they’ve left that he realises that one of the witch’s prophecies has come true so he starts to wonder about the second one. Act 5 is the last act of the play and in scene 5; there are two main events. Firstly, he is told by Seyton, a messenger, that his wife is dead. After this, Macbeth has a soliloquy. Then, straight away, a messenger hurries in and tells Macbeth that Birnam Woods are advancing. This is a turning point in the play because at this point, Macbeth recalls the witches’ prophecy, which said that he was not die until Birnam Woods advanced. The scene ends with Macbeth ordering to ring alarms. The main characters in Act 1, Scene 3 are the witches, Macbeth and Banquo. The witches seem like a big contradiction. The first witch told a story about torturing a sailor to death because his wife refused to give her chestnuts! This shows just how evil the witches were. However, they called themselves sisters and they worked together. For example, the second and third witches offer to give the first witch wind to help. â€Å"I’ll give thee a wind† (line 10). This is contradictory because they have a good side and a bad side so it makes the audience ask themselves if the witches can be trusted. It turns out, at the end of the play, that although the witches told the truth, a lot of trouble could have been avoided if they hadn’t intervened because Macbeth would not have had the ambition to be the king. This is the first scene in which we see Macbeth on stage. So far, he has been portrayed as a brave soldier and â€Å"noble Macbeth†. However, when he sees the witches, he becomes scared. We know Macbeth doesn’t want to admit that he is scared because he doesn’t say it. However, we can tell because in line 49, Banquo says, â€Å"why do you start and seem to fear / things that do sound so fair?† Banquo doesn’t believe the witches at first. He doesn’t take them seriously at all and once they have vanished, he asks himself if him and Macbeth had taken drugs: â€Å"or have we eaten on the insane root,† (line 82). When he finds out that Macbeth is actually Thane of Cawdor, he is really surprised and in line 105, says, â€Å"what, can the devil speak true?† The â€Å"what,† shows the audience that Banquo is surprised. Macbeth is the only major character in act 5, scene 5. However, he changes a lot throughout the scene. At the beginning, he is extremely confident. He says, in lines 2 to 3 that his â€Å"castle’s strength / will laugh a siege to scorn†. After he hears the cry from offstage, he talks to make the audience feel sorry for him. â€Å"I have supped full with horrors,† (line 13) means that he has had enough horror so he is immune to them. He seems numb to the Queen’s death. This is really odd because at the beginning of the play, he seems truly in love with his wife. He only mentions three lines about his wife. After this, he starts talking about life. When the messenger tells him the news about Birnam Woods, he becomes really rude and calls him a â€Å"liar and slave!† Very soon after, he appears much less confident than at the beginning of the scene. He even tells the messenger that he can treat him like he would treat the messenger, which is to hang hi m on a tree until he dies of hunger. Line 39, â€Å"if thy speech be sooth / I care not if thou dost for me as much† The witches in Act 1, Scene 3 all speak in verse; the rhythm is steady and they speak in rhyming couplets. This shows that they are different and odd to normal people. They also speak to Macbeth and Banquo in riddle. This gives the audience doubts about how trustworthy the witches are. In fact, this turns out to be the main problem in the play because a lot would have been avoided if Macbeth had known the whole and exact truth. As soon as Macbeth arrives, he asks lots of questions. He seems desperate to know more and in his speech in line 68, he pleads the witches to â€Å"tell him more†. He also says everything that is going through his head out loud. This shows the audience that he is confused. In this speech, he also uses a lot of commanding words such as â€Å"say†¦ I charge you†¦ you owe†¦ speak†¦ tell†. After the witches vanish, Macbeth starts speaking in really short sentences, which make him seem more thoughtful because we don’t know w hat he’s thinking in between his sentences. After Ross and Angus have left, Macbeth says a lot to the audience (soliloquies). This shows his thoughts and gives the audience audience privilege. This makes the plot seem more exciting. He asks himself a lot of questions in these speeches, which is a change from the constant commanding tone he was using earlier. Lines 131-135, he says, â€Å"why hath it given me earnest of success† meaning, ‘why was I promised a promotion (to be the Thane of Cawdor)’ then he goes on to ask himself â€Å"why do I yield to that suggestion†¦Ã¢â‚¬  meaning, ‘why does it give me such horrible thoughts?’ The audience do not know what these thoughts are although later, we find out that he couldn’t help thinking of murdering King Duncan. At one point, Macbeth even goes into a daze! We know this because in line 142, Banquo says, â€Å"look how our partners rapt.† At the beginning of Act 5, Scene 5, Macbeth gives a lot of orders (Shakespeare used lots of exclamation marks when writing the play). These show that Macbeth is very confident. In lines 2-4, Macbeth says, â€Å"Our castle’s strength will laugh a siege to scorn†. This shows he is not just confident in his way of saying things, but also very arrogant and confident that he will win. However, after his wife, the Queen, dies, Macbeth’s confidence drops. His soliloquy makes the play less realistic because people don’t usually talk to themselves however it is still evocative because it clearly shows Macbeth’s emotions. In his soliloquy, Macbeth mentions that his wife should have died at a more convenient time; line 17, â€Å"she should have died hereafter.† This shows that Macbeth is suddenly completely numb in contrast to him being deeply in love before his wife died. Perhaps, at this pint he is telling himself that he didn’t really care abo ut the Queen because he didn’t want to show his being upset. In line 20, Macbeth starts talking about the meaning of life. This is showing he has a soft side and it’s also the point at which, in my opinion, he really starts to feel sorry for himself and realises the damage of all his mistakes. This section could also be a metaphor because he is comparing life to a play. He reminds the audience that the play is just an unreal story by saying that it’s just a story â€Å"told by an idiot†. It may be that Shakespeare is calling himself an idiot but I think this is highly unlikely. I think, on the other hand, that Macbeth is calling the author (Shakespeare) an idiot because Macbeth’s part in the play is not nice (he turns out to be the tragic hero). Moreover, by saying that an actor plays his character â€Å"and then is heard no more†, Macbeth hints at the end of the story at which he dies. Also, he is near the end of his acting and will probably be seen no more. He is emphasizing this point; perhaps to let people remember him once the play is over. It seems as though Shakespeare wrote this as Macbeth’s true moments, by himself, on stage as the hero. Macbeth’s evocative soliloquy, which shows his soft side, contrasts his really rude manner later. For example, in line 34, he shouts â€Å"liar and slave†. Then he quickly becomes less scary as he starts doubting himself and he even tells the messenger that he has the permission to hand Macbeth onto â€Å"a tree till he dies of hunger†. These sudden changes in moods and politeness show that Macbeth has confused emotions. Equivocation and Evil are the two main themes in Act 1, Scene 3. Equivocation is also the main theme throughout the play. In scene 3, the witches all talk in riddle. This is unclear and therefore a form of equivocation because if the witches told the complete truth, the plot would have been completely different. For example Macbeth would have understood the Birnam Woods riddle. He would have understood that the ‘forest advancing’ meant an army of soldiers against him. Evil is the other main theme in Act 1, Scene 3. This is the scene in which Macbeth contemplates killing King Duncan. He describes his thoughts as awful and they can’t even be spoken; lines 138-139 â€Å"Whose horrid image doth unfix my hair / and make my seated heart knock at my ribs.† However, once Duncan is dead, Macbeth is suddenly able to kill more people more easily. So, what originally seemed very evil became normal for Macbeth. The Evil theme continues in act 5, scene 5. For example in lines 3-5, Macbeth says â€Å"here let them lie till famine and the ague eat them up† Macbeth said this without second thoughts showing that he does not feel very disturbed about people being killed or dieing horribly. Also, the evil in this scene is based more on Violence and Tyranny. For example, in lines 37-39, Macbeth says â€Å"if though speak’st false / upon the next tree shall hang though alive / Till famine cling thee†. This is clearly more gruesome and evil than killing Duncan with a dagger but it is given less importance because killing, by this point, has become more accepted in the play. Another theme in act 5, scene 5 is Appearance Vs Reality. For example, it appears that Birnam Woods are advancing and Macbeth doesn’t understand this however, it turns out to an army of soldiers carrying logs. We can tell that no body really understood why the woods were advancing because in lines 32-33 , a messenger says, â€Å"and anon, methought / the wood began to move†. Also, it appears, when Macbeth is talking about the Queen dieing, that he doesn’t care and he seems completely numb to any pain. However, I think Macbeth is just putting on a false appearance at this point to pretend he is not upset and he’s strong but in fact, I think the reality is that he is very upset but hiding it. Also, going by the riddles, it appears Macbeth is not going to die. He was promised by the second apparition in act 4, scene 1 (line 82-83) that â€Å"none of woman born / shall harm Macbeth.† Both Macbeth and the audience think that everybody is ‘of woman born’ and therefore Macbeth will not die. However, the reality, as is revealed in the very last scene in lines 15-16 in which Macduff explains that he, â€Å"was from his mother’s womb / Untimely ripped.† Meaning that he was born by caesarean section and is therefore going to kill Macbeth. Throughout the whole play, the staging is important to pass messages on to the audience. In Act 1, Scene 3, the witches enter with thunder. This emphasizes the fact that they are weird and mysterious. Also, thunder is usually thought of as a negative thing so Shakespeare, by making the witches related to thunder, give the audience a bad impression of them. It’s as though the thunder symbolises the witches’ wickedness. Also in line 33, the witches dance together â€Å"hand in hand† and they call themselves â€Å"The weird witches† showing that they are different to normal people. The audience sees the witches as outsiders and obviously would find it hard to relate to them. Another weird and mysterious part is when the witches vanish after line 78. The withes actions show the audience that they are different and this, consequently, gives the audience doubts about whether they can be trusted as, as humans, we naturally find it difficult to trust people/things we don’t know. Contrary to the witches, Ross and Angus are completely normal people, and in fact, they enter normally, with no special staging effects. Macbeth is given speaches a couple of times during Act 1, Scene 3. At first, he speaks aside in line 117-119 when he marvels at being Thane of Cawdor and Thane of Glamis and he knows that being king is still to come. He also speaks aside in lines 130, 133, 147 and 152. All of these aside speeches make the play more unrealistic. Shakespeare did this because with the play being unrealistic, it can represent the characters emotions more effectively so they play is more evocative. It also means that the audience solely concentrate on the characters and their emotions. This emphasizes them so this play is written about the emotions and the plot is just a way of portraying them. In act 5, scene 5 the Queen dies offstage. This draws the audience’s feelings towards Macbeth’s emotions and feelings as opposed to the actual death itself. This is important because death has become relatively insignificant by this point in the play because so many people have died. Therefore, drawing attention to the death would not be very effective. However, the audience is very interested in how Macbeth has changed throughout the play so his soliloquies and attention to him are very important in this part of the play. In the 17th century, Shakespeare’s plays were more enjoyed by the public because at the time, they also had a social significance. Historians have also been able to learn about life during the 17th century through Shakespeare’s play as they are closely related to society at that time. For example, in the 1600s there was a big concern about witches. People were extremely scared of them and there were rewards for catching them and horrible punishments for being a witch. This contrasts the play as the witches actually told the truth. The audience is as surprised as Macbeth is when he finds out that he was the Thane of Cawdor. In line 108, Macbeth says, â€Å"what, can the devil speak true?† to show his surprise. On the other, it was nasty of the witches to interfere and they were the main cause of all the problems so they proved to be as bad as society thought they were. The chestnut story in Act 1, Scene 3 shows the audience just how evil the witches are. This makes the audience feel happy because they have just been proved right about their thoughts about witches. Moreover, during Shakespeare’s time, loyalty to the king was one of the most important things. Therefore, Macbeth’s thoughts (line 33) were extremely bad. It would have shocked the audience. This is also one of the main reasons that Duncan’s death is offstage. It would have offended the King if he were shown dieing on stage. In Act 5, Scene 5, it is especially surprising that Macbeth tells the messenger that he is allowed to hang him alive from a tree and let him die of hunger because during the 17th century, social status was very important and the King’s status would have been considered to be much greater than a messenger’s. The fact that Macbeth told the messenger that he could treat him the same, means that Macbeth is bringing his status right down much lower than a King would normally be; to the level of the messenger. In society at that time, this would be extremely different to what everyone was used to so this event would have a huge impact on the audience. In my opinion, the unreality of the play is the main reason that Shakespeare managed to appeal to his audience both in the 17th century and in the modern era. The plot obviously had more social significance in the 17th century and was therefore probably more effective then but the plot isn’t the most important part whereas the emotions our. As our emotions haven’t changed, the play can still be enjoyed now and the modern audience can still understand, and be affected by the evocative speeches and scenes. Cami Rothe Form: 10PE Teacher: A. Partington Coursework: Pre 1914 Drama / Shakespeare

Why Cow’s Milk is a Human Health Risk

Except for animals under human influence and Western gulls who steal milk from lactating seals, humans are the only known species that drinks the breast milk of another species, and the only known species that continues to drink breast milk into adulthood. Needing Milk Milk from a cow is as necessary as milk from a pig or a horse or a giraffe. Human breast milk is the perfect food for human babies, while cow’s milk is the perfect food for baby cows. Cow’s milk naturally contains a large amount of hormones and protein needed to turn an 80-pound calf into a 1,000-pound cow in one year. That amount of protein and hormones is not only unnecessary but unhealthy for humans. Because they occur naturally, these hormones are even found in organically produced milk. The Harvard School of Public Health and Harvard Medical School are downright critical of the USDAs recommendation of dairy products at every meal. Harvard states, there is little evidence that high dairy intake protects against osteoporosis but substantial evidence that high intake can be harmful. If dairy is so bad, why does the USDA recommend so much dairy? Harvard blames industry influences, stating that their recommended diet is based exclusively on the best available science and was not subjected to political and commercial pressures from food industry lobbyists. The American Dietetic Association supports a dairy-free, vegan diet: It is the position of the American Dietetic Association that appropriately planned vegetarian diets, including total vegetarian or vegan diets, are healthful, nutritionally adequate, and may provide health benefits in the prevention and treatment of certain diseases. Besides containing saturated fats, cholesterol, hormones, and too much protein, milk is also linked to testicular cancer, breast cancer, and prostate cancer. Fat, Cholesterol and Protein Many dairy products tend to be high in saturated fats and cholesterol, which have been linked to heart disease. The American Dietetic Association states: Features of a vegetarian diet that may reduce risk of chronic disease include lower intakes of saturated fat and cholesterol and higher intakes of fruits, vegetables, whole grains, nuts, soy products, fiber, and phytochemicals. Milk protein is also a concern, and the protein in milk has been linked to coronary deaths and to hardened, narrowed arteries. Hormones, and Cancer In 2006, a researcher from the Harvard School of Public Health found a strong link between dairy consumption and hormone-dependent cancers; testes, breast, and prostate. Scientist/physician Ganmaa Davaasambuu believes that the naturally occurring hormones in a pregnant cow’s milk increase the risks for these types of cancer. Milk from cows contains â€Å"considerable amounts of female sex hormones,† accounting for 60% to 80% of estrogens consumed by humans. Although the research focused on dairy, Ganmaa’s findings implicated a variety of animal products, as well as dairy: Butter, meat, eggs, milk, and cheese are implicated in higher rates of hormone-dependent cancers in general, she said. Breast cancer has been linked particularly to consumption of milk and cheese. Ganmaa’s findings are not unique. According to dietician George Eisman, in the US, one in six men gets prostate cancer. Only one in 200,000 men gets prostate cancer in China, where dairy is not regularly consumed. Also according to Eisman, breast cancer is highest in the countries with the highest dairy consumption. A study in England found that even within England, the counties with the highest dairy consumption had the highest rates of breast cancer. Eisman states that consuming dairy is â€Å"the most abnormal, crazy thing that we do.† Contaminants in Milk Contaminants in milk are another serious concern. American milk is banned in the European Union because of added recombinant bovine growth hormone (rBGH). When administered to cows, rBGH causes the cows to produce up to 20% more milk but also causes the cows to produce more Insulin-like Growth Factor 1 (IGF-1). According to the Organic Consumers Association, some of the rBGH given to cows ends up in the milk. The Cancer Prevention Coalition (CPC) states: It is highly likely that IGF-1 promotes transformation of normal breast cells to breast cancers. In addition, IGF-1 maintains the malignancy of human breast cancer cells, including their invasiveness and ability to spread to distant organs. RBGH also increases the risk of mastitis, which sometimes leads to pus, bacteria, and blood getting into the milk. Federal law in the US allows up to 50 million pus cells per cup of milk. If rBGH is so dangerous and is banned in the EU, why is it legal in the US? The CPC believes that â€Å"Monsanto Co., the manufacturer of rBGH, has influenced U. S. product safety laws permitting the sale of unlabeled rBGH milk.† Another contaminant found in cow’s milk is pesticide residues. Residues are fat-soluble, which means they become concentrated in the milk and tissues of animals. Calcium While cow’s milk is high in calcium, it is also high in protein. Excess protein in our diets causes calcium to leach out of our bones. Dr. Kerrie Saunders states, â€Å"North America has one of the highest consumptions of dairy products, and also the highest incidence of osteoporosis.â€Å" To combat osteoporosis, Saunders recommends exercise and â€Å"beans and greens† for a source of calcium that is not excessively high in protein. Ganmaa also recommends getting calcium from green leafy vegetables. Furthermore, calcium intake may be less important for bone health than we’ve been led to believe. A study by researchers from the Harvard School of Public Health published in 1997 found that increased consumption of milk and other calcium-rich foods by adult women did not reduce the risk of osteoporotic bone fractures. Calcium retention is also important for preventing osteoporosis. Sodium, smoking, caffeine and physical inactivity can all cause us to lose calcium. While animal rights advocates are vegan for ethical reasons, it’s important to know that cow’s milk is not necessary for human health and foregoing dairy may have health benefits.