Vitamin Supplements What Do Most Americans Need Essays - Vitamins

Vitamin Supplements: What Do Most Americans Need? Vitamin supplements: What do most Americans need? Vitamin supplements: What do most Americans need? That is the question I will be attempting to answer in the following few pages. To start, I will talk about their beginning as well as their recent growth in popularity in the past decade. I will discuss the medical views that are for and against the use of supplements, what types are most important, and what types people need. To conclude, I will tell about the supplements that I take and whether I will continue to do so. Many years ago, there existed diseases such as scurvy, rickets, and everyday colds. These illnesses were the cause of a lot of sickness and sometimes led to other illness and death. Chemists looked everywhere for a cure. Then came the first supplement to become popularized in its uses this was vitamin C. This wonder pill was said to help the body fight colds and other sickness. With the discovery and use of these new vitamins the Food and Nutrition Board convened in 1941 to determine adequate dietary intakes of essential nutrients. At this time deficiency diseases were a common public-health problem. Since that meeting, with the process of food fortification and improved diets, these diseases have been relatively uncommon except in the poor and elderly. In 1943, the first Recommended Dietary Allowances (RDAs) were published. (Skerrett, 25) These amounts were stated in what amounts were just enough to maintain good health, as that any more would just be excreted by the body or build u p in the tissues. Vitamin supplements have grown in popularity at a pretty constant rate since their development, but have become especially popular in recent years. The baby boomer generation that grew up on healthy diets is finding it hard to maintain that healthy lifestyle, and would rather now just pop a pill. Some experts fear that we will become dependent on these supplements and find everyone taking 50-60 different pills each day. Today, vitamin supplements are a $2 billion a year business in the United States. Even with all these wonder pills available in the market, experts still agree that foods should be the first and most appropriate source of nutrients. (Ross, 354-55) In the past few years, the most popular supplement group has been the antioxidants-beta-carotene associated with vitamin A as well as the antioxidants in vitamin E. Antioxidants in general help clean up toxic products created by normal functions of the body's cells. Cells use oxygen to burn their fuel and the byproducts of this process include free radicals. These are chemical compounds that combine easily with fats, proteins, and other substances in the body. (H.H.L, 1) As far as beta-carotene's abilities and uses go, there have been mixed reports. On one hand, these are being praised as a modern day Fountain of Youth saying that research was showing that it may reduce the risks of some forms of cancer, heart disease, strokes, as well as slow the aging process. The findings showed that supplement takers over the age of 65 scored better on tests of cognitive abilities such as reading and remembering maps. (Godbey, 20) In a test published by the National Cancer Institute persons who received supplements including beta-carotene had a reduced cancer rate of 13 percent below previous levels. (Antioxidants, 2) In another report by the American Heart Association, women who consumed high amounts of antioxidant containing foods had a 33 percent lower risk of heart attack and 71 percent lower risk of stroke than women who did not consume high amounts of beta-carotene. In other studies, low beta -carotene levels in smokers is highly correlated with increased r isk of lung cancer and death from all cancers. Along with these helpful effects, beta-carotene is now being linked with a significantly stronger immune system that reduces infection-related illness by 50 percent. (Skerrett, 27) The other side of the beta-carotene argument is associated to health problems in people who smoke. In a study of 18,000 smokers over a six-year period supplementing with vitamin A and beta-carotene, the cancer rate increased by 17 percent over previous levels. The main focus of the experiment, lung cancer, found an increased amount of 46

Implication Isothermal Amplification Based Methods Detection Infectious Diseases Biology Essay Example

Implication Isothermal Amplification Based Methods Detection Infectious Diseases Biology Essay Example Implication Isothermal Amplification Based Methods Detection Infectious Diseases Biology Essay Implication Isothermal Amplification Based Methods Detection Infectious Diseases Biology Essay Methods for the diagnosing of infective diseases have crawled in the last 2-3 decennaries. Many trials that form the anchor of the modern microbiology research lab are based on really old and labor-intensive engineerings such as microscopy. Invention and execution of PCR had lead to major progresss in country of research and nosologies. However, it requires a thermocycler and longer clip to divide two DNA strands and magnify the needed fragment. Burning demand of the hr include more rapid trials without giving sensitiveness, value-added trials, and point-of-care trials for both high- and low-resource scenes. Over the period of last few old ages, research has been focused on alternate methods to better the diagnosing of infective diseases. These include assorted isothermal amplification-based molecular attacks. In this article, we review these isothermal nucleic acerb elaboration engineerings and their applications along with some of the virtues and demerits of these trials. Keywords: PCR, TMA, NASBA, SDA, LAMP, HAD, cHDA, RCA, SPIA, IMDA, SMART Introduction In vitro Nucleic Acid elaboration was for the first clip described in 1971 ( Kleppe ) . Followed by synthesis of tRNA cistron by primer-directed DNA fix and this was non exponential elaboration. In 1983, Kary Mullis postulated the construct of the polymerase concatenation reaction ( PCR ) but remained theoretical until 1985 when Saiki published the first application of PCR on beta-Globin. Thereafter, polymerase concatenation reaction became anchor of bing research and diagnostic universe. Inventions in biotechnology that combine molecular biological science, microfabrication and bioinformatics are traveling nucleic acerb engineerings from futuristic possibilities to common research lab techniques and manners for disease diagnosings. In this manner, elaboration of nucleic acids is widely used in research, forensics, medical specialty, and agribusiness [ 1 ] . One of the most widely used elaboration methods is the polymerase concatenation reaction ( PCR ) , which is a mark elaboration method [ 2 ] . A PCR reaction typically utilizes two oligonucleotide primers, which are hybridized to the 5and 3ends of the mark sequence, and a Deoxyribonucleic acid polymerase, which can widen the annealed primers by adding on deoxyribonucleoside-triphosphates ( dNTPs ) to bring forth double-stranded merchandises.By raising and take downing the temperature of the reaction mixture, the two strands of the DNA merchandise are separated and can function as templets for the following unit of ammunition of tempering and extension, and the procedure is repeated [ 3 ] . Although PCR has been widely used by research workers, but it is labour intensive, requires expensive thermocycling machine and expertness. Several isothermal elaboration techniques have been developed in the last two decennaries without utilizing thermocycler machine. These non-PCR based methods have exploited the high fidelity polymerase of phages and some accessary proteins for in vitro nucleic acid elaboration. All these methods do non necessitate temperature cycling, operate at a changeless temperature, and offer possible advantages including cost, velocity, portability and decreased sensitiveness to inhibitors over PCR. We here describe the best known isothermal elaboration methods ( such as written text mediated elaboration ( TMA ) or self-sustained sequence reproduction ( 3SR ) , nucleic acid sequence-based elaboration ( NASBA ) , strand displacement elaboration ( SDA ) , loop-mediated isothermal elaboration of DNA ( LAMP ) , helicase-dependent elaboration ( HDA ) , round helicasedependent elaboration ( cHDA ) ) , turn overing circle elaboration ( RCA ) , individual primer isothermal elaboration ( SPIA ) , signal mediated elaboration of RNA engineering ( SMART ) and isothermal multiple supplanting elaboration ( IMDA ) and their applications in molecular diagnosing. 1. TRANSCRIPTION MEDIATED AMPLIFICATION ( TMA ) Transcription mediated elaboration ( TMA ) is RNA written text elaboration system which uses RNA polymerase ( T7 RNA polymerase ) to do RNA from a RNA booster sequences engineered in the primer part, a contrary RNA polymerase ( M-MuLV ) to bring forth complementary Deoxyribonucleic acid from the RNA templets ( Guatelli et al, 1990 ) . This RNA elaboration engineering has been farther improved by presenting a 3rd enzymatic activity, Rnase H, to take the RNA from complementary DNA without the heat-denatured measure ( Fig No.2 ) . Therefore, the thermocycling measure has been eliminated, bring forthing an isothermal elaboration method named self-sustained sequence reproduction ( 3SR ) [ 5 ] . It is single-tube reaction, amplifies either DNA or RNA, and produces RNA amplicons, in contrast to most other nucleic acerb elaboration methods that merely bring forth DNA. It has really rapid dynamicss ensuing in a billion fold elaboration within 15-30 proceedingss. The terminal merchandises of TMA can be detected utilizing gel cataphoresis, fluorescence investigations and colorimetric check. TMA has been used for sensing of N.gonorrhoeae and C. trachomatis ( Hobbs et.al.2008 ) HCV Ferraro et Al. 2008, Gelderblom et al.2007, West Neil febrility ( Ziermann et.al. 2008 ) . 2. NUCLEIC ACID SEQUENCE BASED AMPLIFICATION ( NASBA ) NASBA was developed by J. Compton in 1991who defined it as a primer-dependent engineering that can be used for the uninterrupted elaboration of nucleic acids in a individual mixture at one temperature. NASBA is peculiarly suited to sensing of genomic, ribosomal or messenger RNA. The merchandise of NASBA is ss RNA of the original mark. 108 fold elaboration of the mark may be obtained in 30 min. It utilises activity of 3 enzymes RNA polymerase ( T7 RNA polymerase ) ( RNA dependant RNA Polymerase activity ) to do RNA from a RNA booster sequences engineered in the primer part, a contrary RNA polymerase ( AMV ) to bring forth complementary Deoxyribonucleic acid from the RNA templets and RNAse H to degrade RNA from DNA-RNA loanblend followed by formation of complementary DNA by rearward RNA polymerase enzyme. Again RNA polymerase ( DNA dependant RNA polymerase activity ) to do RNA transcripts from double stranded hence functional T7 RNA booster sequences ( Fig No. 3 ) . The terminal merchandises of NASBA can be detected utilizing gel cataphoresis, fluorescence investigations ( NASBA existent clip ) and colorimetric check ( NASBAELISA ) [ 6-8 ] . Food and Drug Administration office of United States of America ( FDA ) has approved the technique in NucliSence preparation ( NASBAECL ) for molecular sensing of some micro-organisms such as HCV and HIV-1 [ 9, 10 ] . Around 500 articles for designation and sensing of micro-organism using NASBA have been reported. 3. STRAND DISPLACEMENT AMPLIFICATION ( SDA ) Strand-displacement elaboration ( SDA ) is an isothermal technique foremost introduced by Walker et Al. 1992. It combines the ability of a limitation endonuclease to nick the unmodified strand of its mark Deoxyribonucleic acid and the action of an exonuclease-deficient Deoxyribonucleic acid polymerase to widen the 3 terminal at the dent and displace the downstream DNA strand. The displaced strand serves as a templet for an antisense reaction and frailty versa, ensuing in exponential elaboration of the mark DNA ( Figure 5 ) . In the originally designed SDA, a mark DNA sample is heat denatured. Four primers ( B1, B2, S1, and S2 ) , present in surplus, and bind the mark strands at places flanking the sequence to be amplified. Primers S1 and S2 have HincII acknowledgment sequences ( 5 GTTGAC 3 ) located 5 to the mark complementary sequences. The four primers are at the same time extended by exo- klenow utilizing dGTP, dCTP, TTP, and dATP ( I ±S ) . Extension of B1 displaces the S1 primer extension merchandise, S1-ext. Likewise, extension of B2 displaces S2-ext. B2 and S2 bind to displaced S1-ext. B, and S1 bind to displaced S2-ext. Extension and displacement reactions on templets S1-ext and S2-ext green goods two fragments with a hemiphosphorothioate HincII at each terminal and two longer fragments with a hemiphosphorothioate HincII site at merely one terminal. HincII nicking and exo-kle now extension/displacement reactions initiate at these four fragments, automatically come ining the SDA reaction rhythm. These reaction stairss continuously rhythm during the class of elaboration. Present in surplus are two SDA primers ( S1 and S2 ) . The 3end of S1 binds to the 3end of the displaced mark strand T1, organizing a semidetached house with 5overhangs. Likewise, S2 binds T2. The 5overhangs of S1 and S2 contain the HincII acknowledgment sequence ( 5- GTTGAC -3 . Exo- klenow extends the 3ends of the semidetached houses utilizing dGTP, dCTP, TTP, and dATP ( I ±S ) , which produces hemiphosphorothioate acknowledgment sites on S1: T1 and S2: T2. HincII nicks the unmodified primer strands of the hemiphosphorothioate acknowledgment sites, go forthing integral the modified complementary strands. Exo- klenow extends the 3end at the dent on S1: T1 and displaces the downstream strand that is tantamount to T2. Likewise, extension at the dent on S2: T2 consequences in supplanting of T1. Nicking and polymerization/displacement stairss rhythm continuously on S1: T1 and S2: T2 because extension at a nick regenerates a nickable HincII acknowledgment site. Target elaboration is exponential because strands displaced from S1: T1 serve as mark for S2 while strands displaced from S2: T2 serve as mark for S1 [ 16 ] . SDA engineering has been used chiefly for clinical diagnosing of infective diseases such as chlamydia and gonorrhoea [ 17-20 ] . This technique can be used for isothermal elaboration of RNA templets in RT-SDA format by adding rearward RNA polymerase to the original procedure [ 21, 22 ] . SDA has been performed on C. trachomatis Verteramo et. Al. 2009, N. gonorrhoeae and C.trachomatis Van et Al. 2001 Cosentino, et Al. 1999, M. TB Hellyer, et Al. 1999. 4. LOOP-MEDIATED ISOTHERMAL AMPLIFICATION ( LAMP ) LOOP-MEDIATED ISOTHERMAL AMPLIFICATION ( LAMP ) is a fresh method that amplify DNA with high specificity, efficiency, celerity at isothermal conditions and relies on car cycling strand supplanting DNA synthesis by Bst DNA polymerase foremost develop by, Notomi, T. et Al. in 2000. It can straight be performed on clinical samples ( DNA Isolation is optional ) . Bst polymerase is thermostable DNA polymerase fromA Bacillus stearothermophilusA ( N3468 ) and contains the 5A? a† Ã¢â‚¬â„¢ 3A? polymerase activity, strand displacement activity but lacks 5A? a† Ã¢â‚¬â„¢3A? exonuclease activity, 3 a† Ã¢â‚¬â„¢5proof reading activity. It uses 4 primers.Two inner primers ( FIP and BIP ) and two outer primers ( F3 and B3 ) [ 29 ] . The elaboration merchandises are stem-loop DNA structures with several upside-down repetitions of the mark and cauliflower-like constructions with multiple cringles ( Figure 7 ) . The LAMP method is besides a extremely efficient elaboration method that allows the synthesis of big sums of Deoxyribonucleic acid in a short clip. As a consequence, pyrophosphate ions are produced in big sums and signifier white precipitates of Mg pyrophosphate. Judging the presence or absence of this white precipitate allows easy differentiation of whether nucleic acid was amplified by the LAMP method [ 30 ] . However the other formats such as gel cataphoresis, real-time turbidimetry, and fluorescence investigations have been used for sensing of LAMP merchandises [ 31, 32 ] . This engineering has widely been used for molecular sensing of several micro-organisms by research workers and it can be a suited pick for design and development of rapid molecular trials in the field [ 33, 34 ] . There has been successful study of 10493 articles for sensing and designation of bacteriums, parasites, viruses and Fungis by LAMP. 5. Rolled CIRCLE AMPLIFICATION ( RCA ) Rolling circle elaboration ( RCA ) generates multiple transcripts of a sequence for the usage in vitro DNA elaboration adapted from in vivo turn overing circle DNA reproduction [ 23, 24 ] . In its original preparation, the RCA reaction involves legion unit of ammunitions of isothermal enzymatic synthesis in which phi 29 DNA polymerase extends a circle-hybridized primer by continuously come oning around the round DNA investigation of several twelve bases to retroflex its sequence over and over once more ( Figure 6 ) [ 25, 26 ] . The individual isolated nature of amplicons in instance of additive RCA may be good for subsequent uses with these DNAs towards their sensing [ 24 ] . This reaction is widely used for diagnostic intents in direct or indirect sensing of different DNA/RNA, protein, and other biomarkers via a set of assorted bimolecular acknowledgment events. A similar reaction was described for RNA polymerases every bit good, but the RNA-generated procedure does non necessitate any hybridization-dependent priming [ 27 ] . Therefore, the latter is merely used to bring forth functional RNA sequences, such as RNA ladders and self-processing ribozymes. Recently, RCA has been farther developed in a technique, named multiply-primed turn overing circle elaboration ( multiply-primed RCA ) that uses the alone belongingss of phi29 DNA polymerase and random primers to accomplish a 10,000-fold elaboration ( Figure 6 ) . The procedure allows elaboration of round DNA straight from cells or plaques, bring forthing, or cloning [ 28 ] . RCA-based attacks have late been pulling attending of diagnostics-oriented biotech companies and research centres for cistron trials and immunochemical assaies, SNP marking and sequencing template readying, single-cell analysis systems, and cistron look surveies [ 26 ] . Johne et Al. 2009 and Rector et al.2004 optimized RCA for Begomovirus and type 16 Human villoma virus severally. 6. HELICASE-DEPENDENT AMPLIFICATION ( HDA ) Helicase-dependent elaboration ( HDA ) is based on the unwinding activity of a DNA helicase [ 41 ] . This procedure uses a helicase, instead than heat, to divide the two strands of a Deoxyribonucleic acid duplex bring forthing single-stranded templets for the intent of in vitro elaboration of a mark nucleic acerb [ 42 ] . Sequence-specific primers hybridize to the templets and are so extended by Deoxyribonucleic acid polymerases to magnify the mark sequence. This procedure repeats itself so that exponential elaboration can be achieved at a individual temperature ( Figure 9 ) . This procedure allows multiple rhythms of reproduction to be performed at a individual incubation temperature, wholly extinguishing the demand for thermo cycling equipment [ 3 ] . The HDA amplicons can be detected utilizing gel cataphoresis, real-time format, and enzyme-linked immunosorbent check ( ELISA ) . [ 41-45 ] HDA has been used for designation of S. aureus, MRSA, Goldmever et al 2008, M. TB Gill, et Al. 2007, H. pylori Gill, et Al. 2007. 7. CIRCULAR HELICASE-DEPENDENT AMPLIFICATION ( cHDA ) round Helicase-dependent elaboration ( cHDA ) is used for magnifying nucleic acids from a round Deoxyribonucleic acid templet. This system combines a Deoxyribonucleic acid polymerase and a helicase readying to magnify a mark sequence every bit good as the full round Deoxyribonucleic acid templet incorporating the mark sequence [ 50 ] . The technique is based on the T7 reproduction machinery, which includes the processive T7 helicase, an exonuclease-deficient T7 DNA polymerase ( T7 sequenase ) and the T7 Gp2.5 single-stranded DNA binding ( SSB ) protein. After the duplex DNA templet is unwound by T7 helicase, specific primers anneal to the separated DNA strands and T7 sequenase extends the 3end of each primer by a rolled circle mechanism to magnify non merely a part defined by the primers but besides uninterrupted concatemers of the templet ( Figure 11 ) . The procedure can be carried out at one temperature ( 25a- ¦C ) for the full procedure. Amplification can be performed utilizing purified plasmid Deoxyribonucleic acid or petroleum cell lysate can magnify inserts every bit big as 10 kilo base braces [ 50 ] . Xu, et al. 2006 performed cHDA on E. coli. 8. SIGNAL MEDIATED AMPLIFICATION OF RNA TECHNOLOGY ( SMART ) SMART is based on the formation of a tripartite junction ( 3WJ ) construction. The method relies on signal elaboration and does non necessitate thermic cycling or affect the copying of mark sequences. The check generates a signal that is extremely target dependant and is appropriate for the sensing of DNA or RNA marks [ 11 ] . It consists of two single-stranded oligonucleotide probes extension investigation and templet investigation, each investigation includes one part that can crossbreed to the mark at next places and another, much shorter, part that hybridizes to the other investigation. The two investigations are annealed to each other in the presence of the specific mark, so organizing a 3WJ ( Figure 4A ) . After 3WJ formation, Bst DNA polymerase extends the short ( extension ) investigation by copying the opposing templet investigation to bring forth a two-base hit stranded T7 RNA polymerase booster sequence ( Figure 4B ) . This dual stranded hence functional booster sequences allows T7 RNA polymerase to adhere and bring forth multiple transcripts of an RNA amplicons and hence being produced merely when a specific mark is present to let 3WJ formation. Each RNA amplicons may itself be amplified by adhering to a 2nd templet oligonucleotide ( investigation for elaboration ) and is extended by DNA polymerase to bring forth a double-stranded booster, taking to written text which increases the RNA amplicons can be detected by an enzyme linked oligosorbent check ( ELOSA ) or in existent clip format [ 12,13 ] . This procedure is in fact a signal elaboration method that the mark sequence is non itself amplified [ 14 ] . SMART has been used for sensing of cyanophages ( Hall, et Al. 2002 ) and E. coli ( Wharam, et Al. 2001 ) . 9. ISOTHERMAL MULTIPLE DISPLACEMENT AMPLIFICATION ( IMDA ) Isothermal Multiple Displacement Amplification ( IMDA ) is based on strand supplanting reproduction of the nucleic acid sequences by multiple primers [ 35 ] . Two sets of primers are used, a right set and a left set ( Figure 8 ) . The primers in the right set are complementary to one strand of the nucleic acid molecule to be amplified and the primers in the left set are complementary to the opposite strand. The 5ends of primers in both sets are distal to the nucleic acerb sequence of involvement where the primers have hybridized to the nucleic acid sequence molecule to be amplified. Amplification is done with the aid of phi29 DNA polymerase is a extremely processive enzyme that incorporates at least 70,000 bases in one binding event entirely with 3-5proof reading and maroon supplanting activity returns by reproduction initiated at each primer and go oning through the nucleic acerb sequence of involvement. A cardinal characteristic of this method is the supplanting of step ining prime rs during reproduction by the polymerase. In another preferable signifier of the method, referred to as whole genome strand supplanting elaboration, a random set of primers is used to randomly prime a sample of genomic nucleic acid [ 36, 37 ] . Amplification returns by reproduction with a extremely processive polymerase initiated at each primer and go oning until self-generated expiration. In this manner, multiple overlapping transcripts of the full genome to be synthesized in a short clip [ 38-40 ] . IMDA has been successfully used for whole genome elaboration from Plasma Lu, et Al. 2005, Blood, bone marrow aspirates, tissue biopsy Luthra, et Al. 2004, Blood Hosono, et Al. 2003. 10. SINGLE PRIMER ISOTHERMAL AMPLIFICATION ( SPIA ) This elaboration engineering uses a individual chimeric primer for elaboration of DNA ( SPIA ) and RNA ( Ribo-SPIA ) [ 46 ] . SPIA employs a individual, target-specific chimeral primer composed of deoxyribonucleotides at the 3- terminal and ribonucleotides at its 5- terminal, RNase H, and a Deoxyribonucleic acid polymerase with a strong strand supplanting activity. Amplification is initiated by crossbreeding the chimeral primer to a complementary sequence in the mark DNA molecule. DNA polymerase initiates primer extension of the hybridized primer and extends along the mark DNA strand. Following induction of the primer extension measure, the 5 RNA part of the drawn-out primer ( RNA-DNA loanblend ) is cleaved by RNase H, therefore liberating portion of the primer-binding site on the mark DNA strand signifier binding of a new chimeral primer. The freshly bound primer competes with the old primer extension merchandise for adhering to the complementary DNA mark sequence and is stabilized by adhering of DNA polymerase and displaces the 5 terminal of the old extension merchandise. As reproduction is once more initiated by primer extension, RNase H cleavage of the 5 RNA part of the freshly extended primer once more frees portion of the primer binding site for subsequent primer binding and reproduction rhythm is repeated. SPIA elaboration can be used for planetary genomic DNA elaboration and for the elaboration of specific genomic sequences and man-made oligonucleotide DNA marks. Ribo-SPIA is likewise suited for planetary and target-specific RNA elaboration ( Figure 10 ) [ 47-49 ] . Ribo-SPIA engineering provides an elegant method for additive, isothermal elaboration of the messenger RNA species in a entire RNA population. Replication is initiated and repeated up to 10,000 times off of each original transcript. Therefore, this procedure can be used for elaboration of big populations of nucleic acid species, which are limited in biological samples, as are normally enc ountered in clinical researches [ 46 ] . Decision In this survey, we described the well studied isothermal engineerings for nucleic acerb elaboration that offer several advantages over PCR in that they eliminate the demand for an expensive and cost-intensive thermocycler. However, these isothermal elaboration engineerings have some limitations that confine their employment in some facets of molecular biological science. For illustration, TMAs requirement is three different enzymatic stairss ( transcription/cDNA synthesis/RNA debasement ) to carry through an isothermal RNA elaboration, [ 4 ] and its get downing stuff is limited to individual stranded nucleic acid/RNA. SDA needs four primers to bring forth initial amplicons and modified deoxynucleotides to supply strand-specific nicking [ 15, 16 ] and it is inefficient at magnifying long mark sequences. LAMP insists upon four to six specific primers that their designs are complicated for new user [ 51 ] . Besides, its concluding merchandise is a complex mixture of stem-loop cauliflower-like DNA constructions of assorted sizes. Nagamine et Al. have devised excess stairss to obtain unvarying single-stranded Deoxyribonucleic acid from LAMP merchandises. This is preferred for assorted hybridisation techniques. The advanced method uses the thermo stable TspRI limitation enzyme to digest elaboration merchandise, [ 52 ] and an extra primer hybridized to the 9-nt 3overhang at the TspRI cleavage site to displace single-stranded Deoxyribonucleic acid by primer extension [ 53 ] . Recently, Kaneko et Al. evaluated the tolerance of LAMP to a civilization medium and some biological substances [ 55 ] . Harmonizing to their survey, the sensitiveness of LAMP was less affected by the assorted constituents of the clinical samples than was PCR ; hence, DNA purification can be omitted. On the other manus, some of these methods such as HDA have a simple reaction strategy, in which a mark sequence can be amplified by two flanking primers, similar to PCR [ 41, 42 ] . One of the most of import advantages of the isothermal elaboration techniques is related to their tolerances to some repressive stuffs that affect the PCR efficiency. Another illustration is about HDA ; a pathogen genomic Deoxyribonucleic acid can even be detected in a human blood sample [ 41 ] . This demonstrates that HDA can be performed on petroleum samples and has the possible to be used as a diagnostic tool. RCA holds a expressed place in DNA nosologies among other isothermal elaboration techniques due to its hardiness and simpleness. As compared with RCA, all other isothermal methods of signal, investigation, or aim DNA elaboration, such as transcription-based system, maroon displacement attack or loop-mediated techniques are instead complicated and in most instances they require anterior assay optimisation [ 26, 54 ] . Although, SDA and RCA are described as isothermal elaboration systems, both methods require an initial heat denaturation measure. Another of import advantage for the isothermal elaboration techniques is no demand to initial heat denaturation at a high temperature followed by elaboration at a lower temperature. This belongings has been reported about some isothermal elaboration methods. For illustration, because there is no necessity for heat denaturation of the templet DNAs, LAMP can be used more easy and quickly in molecular medical specialty [ 56 ] . As DNA helicase can wind off double-stranded mark Deoxyribonucleic acid at the beginning of the reaction, the full HDA reaction can be performed at one temperature [ 41 ] . Beyond the shadow of uncertainty, these isothermal elaboration based engineerings be at discrepancy with their nature and volume of sample required, processing of specimen, and methods of elaboration and sensing. In spite, of these many restrictions isothermal elaborations based techniques would successfully vie with its widely employed thermic cycler based predecessor ( PCR ) for the figure of diagnostic applications. On the whole, simpleness and isothermal nature of these methods offer great potencies for the development of handheld DNA diagnostic devices that could be used to observe pathogens at point-of-care or in the field.

Language Notes on the Use of Aint in English

Language Notes on the Use of Aint in English As far as I know, only one rule of English usage has ever made its way into a childrens jump-rope rhyme: Dont say aint or your mother will faint,Your father will fall in a bucket of paint,Your sister will cry, your brother will die,Your cat and dog will call the FBI. Though frequently heard in casual speech, aint has been described as the most stigmatized word in English. Dictionaries usually label it dialectal or nonstandard, while some purists even deny its right to exist, insisting that aint isnt a word. What is it about this simple negative contraction that agitates language mavens and spreads fear on the playground? As these notes demonstrate, the answer is surprisingly complex. Grammar and Usage[The] two meanings of grammarhow the language functions and how it ought to functionare easily confused. To clarify the distinction, consider the expression aint. Unless used intentionally to add colloquial flavor, aint is unacceptable because its use is considered nonstandard. Yet taken strictly as a part of speech, the term functions perfectly well as a verb. Whether it appears in a declarative sentence (I aint going) or an interrogative sentence (Aint I going?), it conforms to the normal pattern for all verbs in the English language. Although readers may not approve of its use, they cannot argue that it is ungrammatical in such sentences.(Gerald J. Alred, Charles T. Brusaw, and Walter E. Oliu, Handbook of Technical Writing, 10th ed. Bedford/St. Martins, 2012) A Brief History of AintAint has had an unusual history. Its a shortened form of several wordsam not, are not, is not, has not and have not. It appears in written English in the 18th century in various plays and novels, first as ant and then as aint. During the 19th century, it was widely used in representations of regional dialect, especially Cockney speech in the UK, and became a distinctive feature of colloquial American English. But when we look at who is using the form in 19th-century novels, such as those by Dickens and Trollope, we find that the characters are often professional and upper-class. Thats unusual: to find a form simultaneously used at both ends of the social spectrum. Even as recently as 1907, in a commentary on society called The Social Fetich, Lady Agnes Grove was defending aint I as respectable upper-class colloquial speechand condemning arent I!She was in a rapidly diminishing minority. Prescriptive grammarians had taken against aint, and it would soon become universally condemned as a leading marker of uneducated usage.(David Crystal, The Story of English in 100 Words. St. Martins Press, 2012) Collocational AintIn Present-Day English, aint is stigmatized even though linguistically it is formed by the same rule speakers use to form arent and other nonstigmatized contracted auxiliary verbs. . . . [T]here is nothing linguistically wrong with it; in fact, aint is used by many speakers in certain fixed expressions and to convey a certain rhetorical effect: It aint over yet! You aint seen nothing yet! If it aint broke, dont fix it.(Kristin Denham and Anne Lobeck, Linguistics for Everyone: An Introduction. Wadsworth, 2010 Prissy, Ridiculous, and Unpopular Alternatives to Aint I?As linguistic scholars have frequently pointed out, it is unfortunate that aint I? is unpopular in educated speech, for the phrase fills a long-felt need. Am I not? is too prissy for down-to-earth people; amnt I? is ridiculous; and arent I?, though popular in England, has never really caught on in America. With a sentence like the one under discussion [Im your best friend, aint I?] you are practically in a linguistic trapthere is no way out unless you are willing to choose between appearing illiterate, sounding prissy, or feeling ridiculous.(Norman Lewis, Word Power Made Easy. Simon Schuster, 1979) Class ActsA correlation exists between the use of aint and social class, i.e. it is more frequent in lower-class speech. In upper-class speech it is indicative of a personal relationship and an informal situation . . . and is employed when the other person knows that the speaker is using aint for stylistic effect, rather than from ignorance or lack of education (Feagin 1979: 217). Since the form is such a strong school-induced shibboleth, informants tend to suppress it in (more formal) interview situations.(Traute Ewers, The Origin of American Black English: Be-Forms in the Hoodoo Texts. Walter de Gruyter, 1995) Gender ActsThere is still in the American popular mind a notion that aint, for all its faults, is masculine, while arent is not simply feminine, but effeminate. In Thomas Bergers novel The Feud (1983), Tony, a high school student, finds that good grammar must take a back seat to his public sexual identity. Tony defends his use of the masculine aint against his girlfriend Eva s objection that it is a sign of ignorance: I dont like to talk like a girl. Somebody might think I was a pansy.(Dennis E. Baron, Grammar and Gender. Yale University Press, 1986) And that aint all. But for now well have to agree with the editors of The American Heritage Book of English Usage: Aint is a word that aint had it easy.

STROKE Assignment Example | Topics and Well Written Essays - 500 words

STROKE - Assignment Example The effect of stroke on a person depends on the region of its occurrence in the brain and the extent of the damage. A stroke can occur to any person. In some affected individuals, there is total recovery while others may have some form of disability. However, a stroke can be prevented. The paper will discuss the signs and symptoms of a stroke. It will also demonstrate how stroke differ from transient ischemic attack (TIA). It will also include tests as well as a type of referrals or consults. There are various signs and symptoms of stroke. The first main symptom for the disease is numbness or weakness. Numbness is evident in the face, arm or leg and usually is felt on one side of the body (National Library of Medicine (NLM), 2015). The other symptom is confusion and challenges on speaking as well as lack of proper understanding of speech. There is also a problem in seeing in one or two eyes. The affected person may also experience challenges in walking, feels dizzy, and may demonstrate loss of balance or coordination (NLM, 2015). The individual may also experience a persistent headache with no identifiable cause. There is a difference between stroke and transient ischemic attack (TIA). In TIA, blood flow to some areas o f the brain is only stopped for a short duration (National Stroke Association, 2014). As it the flow stops temporary, the symptoms may be like of those experienced in stroke. However, the symptoms last only for a day or less (National Stroke Association, 2014). The other difference is that TIA does not lead to any permanent damage in affected person (National Stroke Association, 2014). However, the signs should be a warning of an impending stroke. The initial workup of a patient with a possible stroke is a crucial process. It entails carrying out a physical assessment (NHS, 2014). The doctor will try the best to establish about the symptoms. Also, various tests will be done to help confirm the diagnosis as well as determine the

History of Cinema Essay Example | Topics and Well Written Essays - 250 words

History of Cinema - Essay Example In particular, the history of cinema dates over 100 years, beginning from late 19th century to the current time. Since the evolution of the cinema during the 1890s, it was considered an economical method of providing mass entertainment. During the first three decades of film discovery, motion pictures did not have sound as the pioneers had not yet figured out how to link sound with the motion pictures. Through continual improvements, inventors succeeded in merging motion pictures with sound in 1923; hence, this resulted in the end of the silent era, while marked the beginning of the sound era (Goodwin, 129). The cinema has undergone various stages of development to reach the current 3-Dimensional films, which has been made possible by continuous learning and improvements. This confirms the fact that the history of Cinema has been useful as it has allowed the ultimate transformation of primitive film to a world class and reliable cinema. Without studying the history of the cinema, there would be nothing to improve. Consequently, the film industry would be lagging behind. As such, we must study the history of cinema because it will enlighten us on new ways of establishing better services than are currently being offered. In conclusion, the study of the past assists us to appreciate the present and makes us ready for the

The Victorian ideal Essay Example for Free

The Victorian ideal Essay The Victorian ideal of womanhood is The Angel in the house. How does Dickens handle his female characters in Great Expectations and how do they relate to this ideal?  Ben A 11wd  In Great Expectations, there are a wide variety of curious and eccentric characters, but it seems that overall, there are significantly more curious female characters (Estella, Miss Havisham, Mrs Joe) than there are male characters (e.g. Mr Pumblechook). It is possible that the reason for the high number of strange women characters might be down to one or more personal experiences in Dickens life. Mrs Joe Gargery, for instance, was directly influenced by Dickens mother, Elizabeth Dickens, who sent Charles, at the age of twelve, to work in a shoe polish factory in order to support the family, who at the time were locked in prison due to the tremendous amount of debt Dickens father was in. Mrs Joe is an exaggerated caricature of Elizabeth, and the antithesis of the angel in the house. The phrase The Angel In The House has its roots in a poem written by the British lyricist Coventry Patmore. He believed that his wife Emily was the perfect incarnation of womanhood; i.e. she was beautiful, obedient, polite, a good cook etc, and it was from her influence that the 211 page volume was written in 1854, and from that poem sprung the expression used to describe a proper housewife doing her duties. Although popular, it received a lot of negative press from the more intelligent liberals both then and now, with some views being reflected by people such as Nel Noddings, who claimed that the Angel was infantile, weak and mindless. But surprisingly, Dickens was actually a supporter of the image presented by Patmore, as were the majority of individuals of the time, even if it does appear that he is criticising the angelic ideal with the character of Mrs Joe for instance, as Dickens actually held quite conservative views on the idea. Mrs Joe is possibly the least angelic person in the entire book, primarily as step by step, Dickens parallels her on every single stage with the Angel, to make her as least angelic as he can, or at least, it appears that he does. She carries with her a stick used for whacking, which Dickens ironically christens Tickler, which she uses to attack both Pip and Joe regularly. There is the image of the thimble, which would usually be used by a housewife similar to the one described in Patmores poem in order to safely knit and sew clothes, and yet Mrs Joe uses it to bash Pip on the head, which is almost the exact opposite of its original purpose. Dickens also appears to cover the ideal of women being perfect cooks, usoing the symbolism of Mrs Joe dropping nails from her top pocket into the bread dough as she is kneading it. being housewives, they would go out an earn money for the family.  The women in Great Expectations range through the social classes from the very top to the very base, and yet all of them have severe defects and are not usually very friendly or caring at all. The most evident of these are the characters of Mrs Joe Gargery and Estella, and to a lesser extent, Molly, Jaggers servant. They all come from different backgrounds, and yet one deals out physical abuse to her husband and brother, one treats the main character with neglect and pretends to loathe him out of spite simply because hes madly in love with her, and the other murdered her newborn baby, of which examples can give us quite a good idea of Dickens attitudes to class as it shows that there is no difference in how nice a person is just because of their status in society. Dickens is trying to suggest that the idea that it was the social class that you were brought up in that defined your personality and character in later life was pure speculation and had no basis in truth whatsoever. It is therefore an opinion voicing not only Dickens personal and more subtly hidden opinions on women, but also another expression of his more widely-known opinions on the Social Class system of England in the Victorian era. Dickens had a sense of social justice in that he was a firm believer that the poor of the country were being treated horribly by the people further up the class system, and although he was no radical or revolutionist, he did believe that it was wrong and so voiced his opinions quite openly in his books concerning this issue. He went from working in a factory when he was twelve to a world renowned author in 30 years, and so held these beliefs firmly and was not merely passing abstract comment on these issues.

Tupac Shakur - Me Against the World Essays -- Music

Tupac Shakur - Me Against the World I was 13 years old. Anticipation thrilled my mind and body as the sea air wafted through the window. My best friend and I were nearing the beach and the mini-van we had inhabited for six hours felt as though it was closing in on us. Thoughts of the beach, the simplicity, and the freedom had controlled my mind for weeks, even months prior to our trip to the beach. Harnessing my recent hormone attack was still a challenge and images of tan girls plagued my mind. Then it happened. A song came on the radio that became my motto for the week; however even though that week ended, the artist never left my life. The voice I heard was that of Tupac Shakur and the song was "I Get Around." The lyrics of the song were boastful and egotistical. I loved it. With Tupac’s display of masculinity as my inspiration, my chances of picking up some of those tan girls were great. Unfortunately, I didn’t and my determined mission was in vein; however to this day Tupac’s music, spirit and camaraderie still gives me inspiration. At the end of that wonderful week we were forced to pack up and leave the freedom and the warmth of the beach. With me I took many memories, and a new tee shirt with the slogan "I Get Around" boldly written on the back. Years have passed. My best friend is still my best friend, but we rarely talk. Now, nearly nothing is quite the same. Friendships have passed; I've changed mentally and physically. Through all of the changes, Tupac has always been at my side. He elevates the highs in my life and illustrates the lows. Tupac Shaker is still influencing me, and I still own that tee shirt. I never lost touch with Tupac. I bought the album with "I Get Around," and I bought his preceding albu... ...ac. I'd lay on the warm sand, whip out my jackknife (headphones) and proceed to examine Tupac's music and life, down to its soul. I could rediscover his essence and get past the thuggish symbolic complex that is so often associated with Tupac. Although the shining serpent has departed this earth I am thankful that he brightens my world and eases my pain everyday. I think when my professor gave this assignment; he intended his class to contemplate for weeks about which CD we would listen to while wasting away on a desert island. What my professor didn’t understand was that I’ve been wasting away for years, and I need this CD to survive, stranded or not. Â   Bibliography: Anson, Robert. "To Die Like a Gangsta." Vanity Fair: Mar, 1997: 244-252. Percy, Walker. "The Loss of the Creature." The Message in the Bottle. New York: Farrar, Srauss, Giroux, 1975: 46-63. Tupac Shakur - Me Against the World Essays -- Music Tupac Shakur - Me Against the World I was 13 years old. Anticipation thrilled my mind and body as the sea air wafted through the window. My best friend and I were nearing the beach and the mini-van we had inhabited for six hours felt as though it was closing in on us. Thoughts of the beach, the simplicity, and the freedom had controlled my mind for weeks, even months prior to our trip to the beach. Harnessing my recent hormone attack was still a challenge and images of tan girls plagued my mind. Then it happened. A song came on the radio that became my motto for the week; however even though that week ended, the artist never left my life. The voice I heard was that of Tupac Shakur and the song was "I Get Around." The lyrics of the song were boastful and egotistical. I loved it. With Tupac’s display of masculinity as my inspiration, my chances of picking up some of those tan girls were great. Unfortunately, I didn’t and my determined mission was in vein; however to this day Tupac’s music, spirit and camaraderie still gives me inspiration. At the end of that wonderful week we were forced to pack up and leave the freedom and the warmth of the beach. With me I took many memories, and a new tee shirt with the slogan "I Get Around" boldly written on the back. Years have passed. My best friend is still my best friend, but we rarely talk. Now, nearly nothing is quite the same. Friendships have passed; I've changed mentally and physically. Through all of the changes, Tupac has always been at my side. He elevates the highs in my life and illustrates the lows. Tupac Shaker is still influencing me, and I still own that tee shirt. I never lost touch with Tupac. I bought the album with "I Get Around," and I bought his preceding albu... ...ac. I'd lay on the warm sand, whip out my jackknife (headphones) and proceed to examine Tupac's music and life, down to its soul. I could rediscover his essence and get past the thuggish symbolic complex that is so often associated with Tupac. Although the shining serpent has departed this earth I am thankful that he brightens my world and eases my pain everyday. I think when my professor gave this assignment; he intended his class to contemplate for weeks about which CD we would listen to while wasting away on a desert island. What my professor didn’t understand was that I’ve been wasting away for years, and I need this CD to survive, stranded or not. Â   Bibliography: Anson, Robert. "To Die Like a Gangsta." Vanity Fair: Mar, 1997: 244-252. Percy, Walker. "The Loss of the Creature." The Message in the Bottle. New York: Farrar, Srauss, Giroux, 1975: 46-63.

Chapter 21 Hermione’s Secret

â€Å"Shocking business†¦ shocking†¦ miracle none of them died†¦ never heard the like†¦ by thunder, it was lucky you were there, Snape†¦Ã¢â‚¬  â€Å"Thank you, Minister.† â€Å"Order of Merlin, Second Class, I'd say. First Class, if I can wangle it!† â€Å"Thank you very much indeed, Minister.† â€Å"Nasty cut you've got there†¦ Black's work, I suppose?† â€Å"As a matter of fact, it was Potter, Weasley, and Granger, Minister†¦Ã¢â‚¬  â€Å"No!† â€Å"Black had bewitched them, I saw it immediately. A Confundus Charm, to judge by their behavior. They seemed to think there was a possibility he was innocent. They weren't responsible for their actions. On the other hand, their interference might have permitted Black to escape†¦ They obviously thought they were going to catch Black single-handed. They've got away with a great deal before now†¦ I'm afraid it's given them a rather high opinion of themselves†¦ and of course Potter has always been allowed an extraordinary amount of license by the headmaster –â€Å" â€Å"Ah, well, Snape†¦ Harry Potter, you know†¦ we've all got a bit of a blind spot where he's concerned.† â€Å"And yet — is it good for him to be given so much special treatment? Personally, I try and treat him like any other student. And any other student would be suspended — at the very least — for leading his friends into such danger. Consider, Minister — against all school rules — after all the precautions put in place for his protection — out-of-bounds, at night, consorting with a werewolf and a murderer — and I have reason to believe he has been visiting Hogsmeade illegally too –â€Å" â€Å"Well, well†¦ we shall see, Snape, we shall see†¦ The boy has undoubtedly been foolish†¦.† Harry lay listening with his eyes tight shut. He felt very groggy. The words he was hearing seemed to be traveling very slowly from his ears to his brain, so that it was difficult to understand†¦. His limbs felt like lead; his eyelids too heavy to lift†¦. He wanted to lie here, on this comfortable bed, forever†¦. â€Å"What amazes me most is the behavior of the Dementors†¦ you've really no idea what made them retreat, Snape?† â€Å"No, Minister†¦ by the time I had come 'round they were heading back to their positions at the entrances†¦.† â€Å"Extraordinary. And yet Black, and Harry, and the girl –â€Å" â€Å"All unconscious by the time I reached them. I bound and gagged Black, naturally, conjured stretchers, and brought them all straight back to the castle.† There was a pause. Harry's brain seemed to be moving a little faster, and as it did, a gnawing sensation grew in the pit of his stomach†¦. He opened his eyes. Everything was slightly blurred. Somebody had removed his glasses. He was lying in the dark hospital wing. At the very end of the ward, he could make out Madam Pomfrey with her back to him, bending over a bed. Harry squinted. Ron's red hair was visible beneath Madam Pomfrey's arm. Harry moved his head over on the pillow. In the bed to his right lay Hermione. Moonlight was falling across her bed. Her eyes were open too. She looked petrified, and when she saw that Harry was awake, pressed a finger to her lips, then pointed to the hospital wing door. It was ajar, and the voices of Cornelius Fudge and Snape were coming through it from the corridor outside. Madam Pomfrey now came walking briskly up the dark ward to Harry's bed. He turned to took at her. She was carrying the largest block of chocolate he had ever seen in his life. It looked like a small boulder. â€Å"Ah, you're awake!† she said briskly. She placed the chocolate on Harry's bedside table and began breaking it apart with a small hammer. â€Å"How's Ron?† said Harry and Hermione together. â€Å"He'll live,† said Madam Pomfrey grimly. â€Å"As for you two, you'll be staying here until I'm satisfied you're — Potter, what do you think you're doing?† Harry was sitting up, putting his glasses back on, and picking up his wand. â€Å"I need to see the headmaster,† he said. â€Å"Potter,† said Madam Pomfrey soothingly, â€Å"it's all right. They've got Black. He's locked away upstairs. The Dementors will be performing the kiss any moment now –â€Å" â€Å"WHAT?† Harry jumped up out of bed; Hermione had done the same. But his shout had been heard in the corridor outside; next second, Cornelius Fudge and Snape had entered the ward. â€Å"Harry, Harry, what's this?† said Fudge, looking agitated. â€Å"You should be in bed — has he had any chocolate?† he asked Madam Pomfrey anxiously. â€Å"Minister, listen!† Harry said. â€Å"Sirius Black's innocent! Peter Pettigrew faked his own death! We saw him tonight! You can't let the Dementors do that thing to Sirius, he's –â€Å" But Fudge was shaking his head with a small smile on his face. â€Å"Harry, Harry, you're very confused, you've been through a dreadful ordeal, lie back down, now, we've got everything under control†¦Ã¢â‚¬  â€Å"YOU HAVEN'T!† Harry yelled. â€Å"YOU'VE GOT THE WRONG MAN!† â€Å"Minister, listen, please,† Hermione said; she had hurried to Harry's side and was gazing imploringly into Fudge's face. â€Å"I saw him too. It was Ron's rat, he's an Animagus, Pettigrew, I mean, and –â€Å" â€Å"You see, Minister?† said Snape. â€Å"Confunded, both of them†¦ Black's done a very good job on them†¦.† â€Å"WE'RE NOT CONFUNDED!† Harry roared. â€Å"Minister! Professor!† said Madam Pomfrey angrily. â€Å"I must insist that you leave. Potter is my patient, and he should not be distressed!† â€Å"I'm not distressed, I'm trying to tell them what happened!† Harry said furiously. â€Å"If they'd just listen –â€Å" But Madam Pomfrey suddenly stuffed a large chunk of chocolate into Harry†s mouth; he choked, and she seized the opportunity to force him back onto the bed. â€Å"Now, please, Minister, these children need care. Please leave.† The door opened again. It was Dumbledore. Harry swallowed his mouthful of chocolate with great difficulty and got up again. â€Å"Professor Dumbledore, Sirius Black –â€Å" â€Å"For heaven's sake!† said Madam Pomfrey hysterically. â€Å"Is this a hospital wing or not? Headmaster, I must insist –â€Å" â€Å"My apologies, Poppy, but I need a word with Mr. Potter and Miss Granger,† said Dumbledore calmly. â€Å"I have just been talking to Sirius Black –â€Å" â€Å"I suppose he's told you the same fairy tale he's planted in Potter's mind?† spat Snape. â€Å"Something about a rat, and Pettigrew being alive –â€Å" â€Å"That, indeed, is Black's story,† said Dumbledore, surveying Snape closely through his half-moon spectacles. â€Å"And does my evidence count for nothing?† snarled Snape. â€Å"Peter Pettigrew was not in the Shrieking Shack, nor did I see any sign of him on the grounds.† â€Å"That was because you were knocked out, Professor!† said Hermione earnestly. â€Å"You didn't arrive in time to hear.† â€Å"Miss Granger, HOLD YOUR TONGUE!† â€Å"Now, Snape,† said Fudge, startled, â€Å"the young lady is disturbed in her mind, we must make allowances –â€Å" â€Å"I would like to speak to Harry and Hermione alone,† said Dumbledore abruptly. â€Å"Cornelius, Severus, Poppy — please leave us.† â€Å"Headmaster!† sputtered Madam Pomfrey. â€Å"They need treatment, they need rest –â€Å" â€Å"This cannot wait,† said Dumbledore. â€Å"I must insist.† Madam Pomfrey pursed her lips and strode away into her office at the end of the ward, slamming the door behind her. Fudge consulted the large gold pocket watch dangling from his waistcoat. â€Å"The Dementors should have arrived by now,† he said. â€Å"I'll go and meet them. Dumbledore, I'll see you upstairs.† He crossed to the door and held it open for Snape, but Snape hadn't moved. â€Å"You surely don't believe a word of Black's story?† Snape whispered, his eyes fixed on Dumbledore's face. â€Å"I wish to speak to Harry and Hermione alone,† Dumbledore repeated. Snape took a step toward Dumbledore. â€Å"Sirius Black showed he was capable of murder at the age of sixteen,† he breathed. â€Å"You haven't forgotten that, Headmaster? You haven't forgotten that he once tried to kill me?† â€Å"My memory is as good as it ever was, Severus,† said Dumbledore quietly. Snape turned on his heel and marched through the door Fudge was still holding. It closed behind them, and Dumbledore turned to Harry and Hermione. They both burst into speech at the same time. â€Å"Professor, Black's telling the truth — we saw Pettigrew — he escaped when Professor Lupin turned into a werewolf –â€Å" â€Å"– he's a rat –â€Å" â€Å"– Pettigrew's front paw, I mean, finger, he cut it off –â€Å" â€Å"– Pettigrew attacked Ron, it wasn't Sirius –â€Å" But Dumbledore held up his hand to stem the flood of explanations. â€Å"It is your turn to listen, and I beg you will not interrupt me, because there is very little time,† he said quietly. â€Å"There is not a shred of proof to support Black's story, except your word — and the word of two thirteen-year-old wizards will not convince anybody. A street full of eyewitnesses swore they saw Sirius murder Pettigrew. I myself gave evidence to the Ministry that Sirius had been the Potters' Secret-Keeper.† â€Å"Professor Lupin can tell you –† Harry said, unable to stop himself â€Å"Professor Lupin is currently deep in the forest, unable to tell anyone anything. By the time he is human again, it will be too late, Sirius will be worse than dead. I might add that werewolves are so mistrusted by most of our kind that his support will count for very little and the fact that he and Sirius are old friends –â€Å" â€Å"But –â€Å" â€Å"Listen to me, Harry. It is too late, you understand me? You must see that Professor Snape's version of events is far more convincing than yours.† â€Å"He hates Sirius,† Hermione said desperately. â€Å"All because of some stupid trick Sirius played on him –â€Å" â€Å"Sirius has not acted like an innocent man. The attack on the Fat Lady — entering Gryffindor Tower with a knife — without Pettigrew, alive or dead, we have no chance of overturning Sirius's sentence.† â€Å"But you believe us.† â€Å"Yes, I do,† said Dumbledore quietly. â€Å"But I have no power to make other men see the truth, or to overrule the Minister of Magic†¦.† Harry stared up into the grave face and felt as though the ground beneath him were falling sharply away. He had grown used to the idea that Dumbledore could solve anything. He had expected Dumbledore to pull some amazing solution out of the air. But no †¦ their last hope was gone. â€Å"What we need,† said Dumbledore slowly, and his light blue eyes moved from Harry to Hermione, â€Å"is more time.† â€Å"But –† Hermione began. And then her eyes became very round. â€Å"OH!† â€Å"Now, pay attention,† said Dumbledore, speaking very low, and very clearly. â€Å"Sirius is locked in Professor Flitwick's office on the seventh floor. Thirteenth window from the right of the West Tower. If all goes well, you will be able to save more than one innocent life tonight. But remember this, both of you: you must not be seen. Miss Granger, you know the law — you know what is at stake†¦You — must — not — be –seen.† Harry didn't have a clue what was going on. Dumbledore had turned on his heel and looked back as he reached the door. â€Å"I am going to lock you in. It is –† he consulted his watch, â€Å"five minutes to midnight. Miss Granger, three turns should do it. Good luck.† â€Å"Good luck?† Harry repeated as the door closed behind Dumbledore. â€Å"Three turns? What's he talking about? What are we supposed to do?† But Hermione was fumbling with the neck of her robes, pulling from beneath them a very long, very fine gold chain. â€Å"Harry, come here,† she said urgently. â€Å"Quick!† Harry moved toward her, completely bewildered. She was holding the chain out. He saw a tiny, sparkling hourglass hanging from it. â€Å"Here –â€Å" She had thrown the chain around his neck too. â€Å"Ready?† she said breathlessly. â€Å"What are we doing?† Harry said, completely lost. Hermione turned the hourglass over three times. The dark ward dissolved. Harry had the sensation that he was flying very fast, backward. A blur of colors and shapes rushed past him, his ears were pounding, he tried to yell but couldn't hear his own voice — And then he felt solid ground beneath his feet, and everything came into focus again — He was standing next to Hermione in the deserted entrance hall and a stream of golden sunlight was falling across the paved floor from the open front doors. He looked wildly around at Hermione, the chain of the hourglass cutting into his neck. â€Å"Hermione, what –?† â€Å"In here!† Hermione seized Harry's arm and dragged him across the hall to the door of a broom closet; she opened it, pushed him inside among the buckets and mops, then slammed the door behind them. â€Å"What — how — Hermione, what happened?† â€Å"We've gone back in time,† Hermione whispered, lifting the chain off Harry's neck in the darkness. â€Å"Three hours back†¦Ã¢â‚¬  Harry found his own leg and gave it a very hard pinch. It hurt a lot, which seemed to rule out the possibility that he was having a very bizarre dream. â€Å"But –â€Å" â€Å"Shh! Listen! Someone's coming! I think — I think it might be us!' Hermione had her ear pressed against the cupboard door. â€Å"Footsteps across the hall†¦ yes, I think it's us going down to Hagrid's!† â€Å"Are you telling me,† Harry whispered, â€Å"that we're here in this cupboard and we're out there too?† â€Å"Yes,† said Hermione, her ear still glued to the cupboard door. â€Å"I'm sure it's us. It doesn't sound like more than three people†¦ and we're walking slowly because we're under the Invisibility Cloak — â€Å" She broke off, still listening intently. â€Å"We've gone down the front steps†¦.† Hermione sat down on an upturned bucket, looking desperately anxious, but Harry wanted a few questions answered. â€Å"Where did you get that hourglass thing?† â€Å"It's called a Time-Turner,† Hermione whispered, â€Å"and I got it from Professor McGonagall on our first day back. I've been using it all year to get to all my lessons. Professor McGonagall made me swear I wouldn't tell anyone. She had to write all sorts of letters to the Ministry of Magic so I could have one. She had to tell them that I was a model student, and that I'd never, ever use it for anything except my studies†¦ I've been turning it back so I could do hours over again, that's how I've been doing several lessons at once, see? But†¦ â€Å"Harry, I don't understand what Dumbledore wants us to do. Why did he tell us to go back three hours? How's that going to help Sirius?† Harry stared at her shadowy face. â€Å"There must be something that happened around now he wants us to change,† he said slowly. â€Å"What happened? We were walking down to Hagrid's three hours ago†¦.† â€Å"This is three hours ago, and we are walking down to Hagrid's,† said Hermione. â€Å"We just heard ourselves leaving†¦.† Harry frowned; he felt as though he were screwing up his whole brain in concentration. â€Å"Dumbledore just said — just said we could save more than one innocent life†¦Ã¢â‚¬  And then it hit him. â€Å"Hermione, we're going to save Buckbeak!† â€Å"But — how will that help Sirius?† â€Å"Dumbledore said — he just told us where the window is — the window of Flitwick's office! Where they've got Sirius locked up! We've got to fly Buckbeak up to the window and rescue Sirius! Sirius can escape on Buckbeak — they can escape together!† From what Harry could see of Hermione's face, she looked terrified. â€Å"If we manage that without being seen, it'll be a miracle!† â€Å"Well, we've got to try, haven't we?† said Harry. He stood up andvpressed his ear against the door. â€Å"Doesn't sound like anyone's there†¦ Come on, let's go.† Harry pushed open the closet door. The entrance hall was deserted. As quietly and quickly as they could, they darted out of the closet and down the stone steps. The shadows were already lengthening, the tops of the trees in the Forbidden Forest gilded once more with gold. â€Å"If anyone's looking out of the window –† Hermione squeaked, looking up at the castle behind them. â€Å"We'll run for it,† said Harry determinedly. â€Å"Straight into the forest, all right? We'll have to hide behind a tree or something and keep a lookout –â€Å" â€Å"Okay, but we'll go around by the greenhouses!' said Hermione breathlessly. â€Å"We need to keep out of sight of Hagrid's front door, or we'll see us! We must be nearly at Hagrid's by now!† Still working out what she meant, Harry set off at a sprint, Hermione behind him. They tore across the vegetable gardens to the greenhouses, paused for a moment behind them, then set off again, fast as they could, skirting around the Whomping Willow, tearing toward the shelter of the forest†¦. Safe in the shadows of the trees, Harry turned around; seconds later, Hermione arrived beside him, panting. â€Å"Right,† she gasped. â€Å"We need to sneak over to Hagrid's†¦. Keep out of sight, Harry†¦.† They made their way silently through the trees, keeping to the very edge of the forest. Then, as they glimpsed the front of Hagrid's house, they heard a knock upon his door. They moved quickly behind a wide oak trunk and peered out from either side. Hagrid had appeared in his doorway, shaking and white, looking around to see who had knocked. And Harry heard his own voice. â€Å"It's us. We're wearing the Invisibility Cloak. Let us in and we can take it off.† â€Å"Yeh shouldn've come!† Hagrid whispered. He stood back, then shut the door quickly. â€Å"This is the weirdest thing we've ever done,† Harry said fervently. â€Å"Let's move along a bit,† Hermione whispered. â€Å"We need to get nearer to Buckbeak!† They crept through the trees until they saw the nervous Hippogriff, tethered to the fence around Hagrid's pumpkin patch. â€Å"Now?† Harry whispered. â€Å"No!† said Hermione. â€Å"If we steal him now, those Committee people will think Hagrid set him free! We've got to wait until they've seen he's tied outside!† â€Å"That's going to give us about sixty seconds,† said Harry. This was starting to seem impossible. At that moment, there was a crash of breaking china from inside Hagrid's cabin. â€Å"That's Hagrid breaking the milk jug,† Hermione whispered. â€Å"I'm going to find Scabbers in a moment –â€Å" Sure enough, a few minutes later, they heard Hermione's shriek of surprise. â€Å"Hermione,† said Harry suddenly, â€Å"what if we — we just run in there and grab Pettigrew –â€Å" â€Å"No!† said Hermione in a terrified whisper. â€Å"Don't you understand? We're breaking one of the most important wizarding laws! Nobody's supposed to change time, nobody! You heard Dumbledore, if we're seen –â€Å" â€Å"We'd only be seen by ourselves and Hagrid!† â€Å"Harry, what do you think you'd do if you saw yourself bursting into Hagrid's house?† said Hermione. â€Å"I'd — I'd think I'd gone mad,† said Harry, â€Å"or I'd think there was some Dark Magic going on –â€Å" â€Å"Exactly! You wouldn't understand, you might even attack yourself! Don't you see? Professor McGonagall told me what awful things have happened when wizards have meddled with time†¦ Loads of them ended up killing their past or future selves by mistake!† â€Å"Okay!† said Harry. â€Å"It was just an idea, I just thought –â€Å" But Hermione nudged him and pointed toward the castle. Harry moved his head a few inches to get a clear view of the distant front doors. Dumbledore, Fudge, the old Committee member, and Macnair the executioner were coming down the steps. â€Å"We're about to come out!† Hermione breathed. And sure enough, moments later, Hagrid's back door opened, and Harry saw himself, Ron, and Hermione walking out of it with Hagrid. It was, without a doubt, the strangest sensation of his life, standing behind the tree, and watching himself in the pumpkin patch. â€Å"It's Okay, Beaky, it's okay†¦Ã¢â‚¬  Hagrid said to Buckbeak. Then he turned to Harry, Ron, and Hermione. â€Å"Go on. Get goin'.† â€Å"Hagrid, we can't –â€Å" â€Å"We'll tell them what really happened –â€Å" â€Å"They can't kill him –â€Å" â€Å"Go! It's bad enough without you lot in trouble an' all!† Harry watched the Hermione in the pumpkin patch throw the Invisibility Cloak over him and Ron. â€Å"Go quick. Don' listen†¦.† There was a knock on Hagrid's front door. The execution party had arrived. Hagrid turned, around and headed back into his cabin, leaving the back door ajar. Harry watched the grass flatten in patches all around the cabin and heard three pairs of feet retreating. He, Ron, and Hermione had gone†¦ but the Harry and Hermione hidden in the trees could now hear what was happening inside the cabin through the back door. â€Å"Where is the beast?† came the cold voice of Macnair. â€Å"Out — outside,† Hagrid croaked. Harry pulled his head out of sight as Macnair's face appeared at Hagrid's window, staring out at Buckbeak. Then they heard Fudge. â€Å"We — er — have to read you the official notice of execution, Hagrid. I'll make it quick. And then you and Macnair need to sign it. Macnair, You're supposed to listen too, that's procedure –â€Å" Macnair's face vanished from the window. It was now or never. â€Å"Wait here,† Harry whispered to Hermione. â€Å"I'll do it.† As Fudge's voice started again, Harry darted out from behind his tree, vaulted the fence into the pumpkin patch, and approached Buckbeak. â€Å"It is the decision of the Committee for the Disposal of Dangerous Creatures that the Hippogriff Buckbeak, hereafter called the condemned, shall he executed on the sixth of June at sundown –â€Å" Careful not to blink, Harry stared up into Buckbeak's fierce orange eyes once more and bowed. Buckbeak sank to his scaly knees and then stood up again. Harry began to fumble with the knot of rope tying Buckbeak to the fence. â€Å"†¦ sentenced to execution by beheading, to be carried out by the Committee's appointed executioner, Walden Macnair†¦Ã¢â‚¬  â€Å"Come on, Buckbeak,† Harry murmured, â€Å"come on, we're going to help you. Quietly†¦ quietly†¦Ã¢â‚¬  â€Å"†¦ as witnessed below. Hagrid, you sign here†¦Ã¢â‚¬  Harry threw all his weight onto the rope, but Buckbeak had dug in his front feet. â€Å"Well, let's get this over with,† said the reedy voice of the Committee member from inside Hagrid's cabin. â€Å"Hagrid, perhaps it will be better if you stay inside –â€Å" â€Å"No, I — I wan' ter be with him†¦. I don' wan' him ter be alone –â€Å" Footsteps echoed from within the cabin. â€Å"Buckbeak, move!† Harry hissed. Harry tugged harder on the rope around Buckbeak's neck. The Hippogriff began to walk, rustling its wings irritably. They were still ten feet away from the forest, in plain view of Hagrid's back door. â€Å"One moment, please, Macnair,† came Dumbledore's voice. â€Å"You need to sign too.† The footsteps stopped. Harry heaved on the rope. Buckbeak snapped his beak and walked a little faster. Hermione's white face was sticking out from behind a tree. â€Å"Harry, hurry!† she mouthed. Harry could still hear Dumbledore's voice talking from within the cabin. He gave the rope another wrench. Buckbeak broke into a grudging trot. They had reached the trees†¦. â€Å"Quick! Quick!† Hermione moaned, darting out from behind her tree, seizing the rope too and adding her weight to make Buckbeak move faster. Harry looked over his shoulder; they were now blocked from sight; they couldn't see Hagrid's garden at all. â€Å"Stop!† he whispered to Hermione. â€Å"They might hear us.† Hagrid's back door had opened with a bang. Harry, Hermione, and Buckbeak stood quite still; even the Hippogriff seemed to be listening intently. Silence†¦ then — â€Å"Where is it?† said the reedy voice of the Committee member. â€Å"Where is the beast?† â€Å"It was tied here!† said the executioner furiously. â€Å"I saw it! Just here!† â€Å"How extraordinary,† said Dumbledore. There was a note of amusement in his voice. â€Å"Beaky!† said Hagrid huskily. There was a swishing noise, and the thud of an axe. The executioner seemed to have swung it into the fence in anger. And then came the howling, and this time they could hear Hagrid's words through his sobs. â€Å"Gone! Gone! Bless his little beak, he's gone! Musta pulled himself free! Beaky, yeh clever boy!† Buckbeak started to strain against the rope, trying to get back to Hagrid. Harry and Hermione tightened their grip and dug their heels into the forest floor to stop him. â€Å"Someone untied him!† the executioner was snarling. â€Å"We should search the grounds, the forest.† â€Å"Macnair, if Buckbeak has indeed been stolen, do you really think the thief will have led him away on foot?† said Dumbledore, still sounding amused. â€Å"Search the skies, if you will†¦. Hagrid, I could do with a cup of tea. Or a large brandy.† â€Å"O' — o' course, Professor,† said Hagrid, who sounded weak with happiness. â€Å"Come in, come in†¦.† Harry and Hermione listened closely. They heard footsteps, the soft cursing of the executioner, the snap of the door, and then silence once more. â€Å"Now what?† whispered Harry, looking around. â€Å"We'll have to hide in here,† said Hermione, who looked very shaken. â€Å"We need to wait until they've gone back to the castle. Then we wait until it's safe to fly Buckbeak up to Sirius's window. He won't be there for another couple of hours†¦. Oh, this is going to be difficult†¦.† She looked nervously over her shoulder into the depths of the forest. The sun was setting now. â€Å"We're going to have to move,† said Harry, thinking hard. â€Å"We've got to be able to see the Whomping Willow, or we won't know what's going on.† â€Å"Okay,† said Hermione, getting a firmer grip on Buckbeak's rope. â€Å"But we've got to keep out of sight, Harry, remember†¦.† They moved around the edge of the forest, darkness falling thickly around them, until they were hidden behind a clump of trees through which they could make out the Willow. â€Å"There's Ron!† said Harry suddenly. A dark figure was sprinting across the lawn and its shout echoed through the still night air. â€Å"Get away from him — get away — Scabbers, come here –â€Å" And then they saw two more figures materialize out of nowhere. Harry watched himself and Hermione chasing afte r Ron. Then he saw Ron dive. â€Å"Gotcha! Get off, you stinking cat –â€Å" â€Å"There's Sirius!† said Harry. The great shape of the dog had bounded out from the roots of the Willow. They saw him bowl Harry over, then seize on†¦. â€Å"Looks even worse from here, doesn't it?† said Harry, watching the dog pulling Ron into the roots. â€Å"Ouch — look, I just got walloped by the tree — and so did you — this is weird?C† The Whomping Willow was creaking and lashing out with its lower branches; they could see themselves darting here and there, trying to reach the trunk. And then the tree froze. â€Å"That was Crookshanks pressing the knot,† said Hermione. â€Å"And there we go†¦Ã¢â‚¬  Harry muttered. â€Å"We're in.† The moment they disappeared, the tree began to move again. Seconds later, they heard footsteps quite close by. Dumbledore, Macnair, Fudge, and the old Committee member were making their way up to the castle. â€Å"Right after we'd gone down into the passage!† said Hermione. â€Å"If only Dumbledore had come with us†¦Ã¢â‚¬  â€Å"Macnair and Fudge would've come too,† said Harry bitterly. â€Å"I bet you anything Fudge would've told Macnair to murder Sirius on the spot†¦.† They watched the four men climb the castle steps and disappear from view. For a few minutes the scene was deserted. Then — â€Å"Here comes Lupin!† said Harry as they saw another figure sprinting down the stone steps and halting toward the Willow. Harry looked up at the sky. Clouds were obscuring the moon completely. They watched Lupin seize a broken branch from the ground and prod the knot on the trunk. The tree stopped fighting, and Lupin, too, disappeared into the gap in its roots. â€Å"If he'd only grabbed the cloak,† said Harry. â€Å"It's just lying there†¦.† He turned to Hermione. â€Å"If I just dashed out now and grabbed it, Snape'd never be able to get it and –â€Å" â€Å"Harry, we mustn't be seen!† â€Å"How can you stand this?† he asked Hermione fiercely. â€Å"Just standing here and watching it happen?† He hesitated. â€Å"I'm going to grab the cloak!† â€Å"Harry, no!† Hermione seized the back of Harry's robes not a moment too soon. Just then, they heard a burst of song. It was Hagrid, making his way up to the castle, singing at the top of his voice, and weaving slightly as he walked. A large bottle was swinging from his hands. â€Å"See?† Hermione whispered. â€Å"See what would have happened? We've got to keep out of sight! No, Buckbeak!† The Hippogriff was making frantic attempts to get to Hagrid again; Harry seized his rope too, straining to hold Buckbeak back. They watched Hagrid meander tipsily up to the castle. He was gone. Buckbeak stopped fighting to get away. His head drooped sadly. Barely two minutes later, the castle doors flew open yet again, and Snape came charging out of them, running toward the Willow. Harry's fists clenched as they watched Snape skid to a halt next to the tree, looking around. He grabbed the cloak and held it up. â€Å"Get your filthy hands off it,† Harry snarled under his breath. â€Å"Shh!† Snape seized the branch Lupin had used to freeze the tree, prodded the knot, and vanished from view as he put on the cloak. â€Å"So that's it,† said Hermione quietly. â€Å"We're all down there†¦ and now we've just got to wait until we come back up again†¦.† She took the end of Buckbeak's rope and tied it securely around the nearest tree, then sat down on the dry ground, arms around her knees. â€Å"Harry, there's something I don't understand†¦. Why didn't the Dementors get Sirius? I remember them coming, and then I think I passed out†¦ there were so many of them†¦.† Harry sat down too. He explained what he'd seen; how, as the nearest Dementor had lowered its mouth to Harry's, a large silver something had come galloping across the lake and forced the Dementors to retreat. Hermione's mouth was slightly open by the time Harry had finished. â€Å"But what was it?† â€Å"There's only one thing it could have been, to make the Dementors go,† said Harry. â€Å"A real Patronus. A powerful one.† â€Å"But who conjured it?† Harry didn't say anything. He was thinking back to the person he'd seen on the other bank of the lake. He knew who he thought it had been†¦ but how could it have been? â€Å"Didn't you see what they looked like?† said Hermione eagerly. â€Å"Was it one of the teachers?† â€Å"No,† said Harry. â€Å"He wasn't a teacher.† â€Å"But it must have been a really powerful wizard, to drive all those Dementors away†¦ If the Patronus was shining so brightly, didn't it light him up? Couldn't you see –?† â€Å"Yeah, I saw him,† said Harry slowly. â€Å"But†¦ maybe I imagined it†¦ I wasn't thinking straight†¦ I passed out right afterward†¦.† â€Å"Who did you think it was?† â€Å"I think –† Harry swallowed, knowing how strange this was going to sound. â€Å"I think it was my dad.† Harry glanced up at Hermione and saw that her mouth was fully open now. She was gazing at him with a mixture of alarm and pity. â€Å"Harry, your dad's — well — dead,† she said quietly. â€Å"I know that,† said Harry quickly. â€Å"You think you saw his ghost?† â€Å"I don't know†¦ no†¦ he looked solid†¦.† â€Å"But then –â€Å" â€Å"Maybe I was seeing things,† said Harry. â€Å"But†¦ from what I could see†¦ it looked like him†¦. I've got photos of him†¦.† Hermione was still looking at him as though worried about his sanity. â€Å"I know it sounds crazy,† said Harry flatly. He turned to took at Buckbeak, who was digging his beak into the ground, apparently searching for worms. But he wasn't really watching Buckbeak. He was thinking about his father and about his father's three oldest friends†¦ Moony, Wormtail, Padfoot, and Prongs†¦. Had all four of them been out on the grounds tonight? Wormtail had reappeared this evening when everyone had thought he was dead†¦ Was it so impossible his father had done the same? Had he been seeing things across the take? The figure had been too far away to see distinctly†¦ yet he had felt sure, for a moment, before he'd lost consciousness†¦. The leaves overhead rustled faintly in the breeze. The moon drifted in and out of sight behind the shifting clouds. Hermione sat with her face turned toward the Willow, waiting. And then, at last, after over an hour†¦ â€Å"Here we come!† Hermione whispered. She and Harry got to their feet. Buckbeak raised his head. They saw Lupin, Ron, and Pettigrew clambering awkwardly out of the hole in the roots. Then came Hermione†¦ then the unconscious Snape, drifting weirdly upward. Next came Harry and Black. They all began to walk toward the castle. Harry's heart was starting to beat very fast. He glanced up at the sky. Any moment now, that cloud was going to move aside and show the moon†¦ â€Å"Harry,† Hermione muttered as though she knew exactly what he was thinking, â€Å"we've got to stay put. We mustn't be seen. There's nothing we can do†¦.† â€Å"So we're just going to let Pettigrew escape all over again†¦Ã¢â‚¬  said Harry quietly. â€Å"How do you expect to find a rat in the dark?† snapped Hermione. â€Å"There's nothing we can do! We came back to help Sirius; we're not supposed to be doing anything else!† â€Å"All right!† The moon slid out from behind its cloud. They saw the tiny figures across the grounds stop. Then they saw movement — â€Å"There goes Lupin,† Hermione whispered. â€Å"He's transforming.† â€Å"Hermione!† said Harry suddenly. â€Å"We've got to move!† â€Å"We mustn't, I keep telling you –â€Å" â€Å"Not to interfere! Lupin's going to run into the forest, right at us!† Hermione gasped. â€Å"Quick!† she moaned, dashing to untie Buckbeak. â€Å"Quick! Where are we going to go? Where are we going to hide? The Dementors will be coming any moment –â€Å" â€Å"Back to Hagrid's!† Harry said. â€Å"It's empty now — come on!† They ran as fast as they could, Buckbeak cantering along behind them. They could hear the werewolf howling behind them†¦. The cabin was in sight; Harry skidded to the door, wrenched it open, and Hermione and Buckbeak flashed past him; Harry threw himself in after them and bolted the door. Fang the boarhound barked loudly. â€Å"Shh, Fang, it's us!† said Hermione, hurrying over and scratching his ears to quieten him. â€Å"That was really close!† she said to Harry. â€Å"Yeah†¦Ã¢â‚¬  Harry was looking out of the window. It was much harder to see what was going on from here. Buckbeak seemed very happy to find himself back inside Hagrid's house. He lay down in front of the fire, folded his wings contentedly, and seemed ready for a good nap. â€Å"I think I'd better go outside again, you know,† said Harry slowly. â€Å"I can't see what's going on — we won't know when it's time –â€Å" Hermione looked up. Her expression was suspicious. â€Å"I'm not going to try and interfere,† said Harry quickly. â€Å"But if we don't see what's going on, how're we going to know when it's time to rescue Sirius?† â€Å"Well†¦ okay, then†¦ I'll wait here with Buckbeak†¦ but Harry, be careful — there's a werewolf out there — and the Dementors.† Harry stepped outside again and edged around the cabin. He could hear yelping in the distance. That meant the Dementors were closing in on Sirius†¦. He and Hermione would be running to him any moment†¦. Harry stared out toward the lake, his heart doing a kind of drumroll in his chest†¦. Whoever had sent that Patronus would be appearing at any moment†¦. For a fraction of a second he stood, irresolute, in front of Hagrid's door. You must not be seen. But he didn't want to be seen. He wanted to do the seeing†¦. He had to know†¦ And there were the Dementors. They were emerging out of the darkness from every direction, gliding around the edges of the lake†¦. They were moving away from where Harry stood, to the opposite bank†¦. He wouldn't have to get near them†¦. Harry began to run. He had no thought in his head except his father†¦ If it was him†¦ if it really was him†¦ he had to know, had to find out†¦. The lake was coming nearer and nearer, but there was no sign of anybody. On the opposite bank, he could see tiny glimmers of silver — his own attempts at a Patronus — There was a bush at the very edge of the water. Harry threw himself behind it, peering desperately through the leaves. On the opposite bank, the glimmers of silver were suddenly extinguished. A terrified excitement shot through him — any moment now — â€Å"Come on!† he muttered, staring about. â€Å"Where are you? Dad, come on –â€Å" But no one came. Harry raised his head to look at the circle of Dementors across the lake. One of them was lowering its hood. It was time for the rescuer to appear — but no one was coming to help this time — And then it hit him — he understood. He hadn't seen his father he had seen himself — Harry flung himself out from behind the bush and pulled out his wand. â€Å"EXPECTO PATRONUM! † he yelled. And out of the end of his wand burst, not a shapeless cloud of mist, but a blinding, dazzling, silver animal. He screwed up his eyes, trying to see what it was. It looked like a horse. It was galloping silently away from him, across the black surface of the lake. He saw it lower its head and charge at the swarming Dementors†¦. Now it was galloping around and around the black shapes on the ground, and the Dementors were falling back, scattering, retreating into the darkness†¦. They were gone. The Patronus turned. It was cantering back toward Harry across the still surface of the water. It wasn't a horse. It wasn't a unicorn, either. It was a stag. It was shining brightly as the moon above †¦ it was coming back to him†¦. It stopped on the bank. Its hooves made no mark on the soft ground as it stared at Harry with its large, silver eyes. Slowly, it bowed its antlered head. And Harry realized†¦ â€Å"Prongs,† he whispered. But as his trembling fingertips stretched toward the creature, it vanished. Harry stood there, hand still outstretched. Then, with a great leap of his heart, he heard hooves behind him. He whirled around and saw Hermione dashing toward him, dragging Buckbeak behind her. â€Å"What did you do?† she said fiercely. â€Å"You said you were only going to keep a lookout!† â€Å"I just saved all our lives†¦Ã¢â‚¬  said Harry. â€Å"Get behind here behind this bush — I'll explain.† Hermione listened to what had just happened with her mouth open yet again. â€Å"Did anyone see you?† â€Å"Yes, haven't you been listening? I saw me but I thought I was my dad! It's okay!† â€Å"Harry, I can't believe it†¦ You conjured up a Patronus that drove away all those Dementors! That's very, very advanced magic.† â€Å"I knew I could do it this time,† said Harry, â€Å"because I'd already done it†¦ Does that make sense?† â€Å"I don't know — Harry, look at Snape!† Together they peered around the bush at the other bank. Snape had regained consciousness. He was conjuring stretchers and lifting the limp forms of Harry, Hermione, and Black onto them. A fourth stretcher, no doubt bearing Ron, was already floating at his side. Then, wand held out in front of him, he moved them away toward the castle. â€Å"Right, it's nearly time,† said Hermione tensely, looking at her watch. â€Å"We've got about forty-five minutes until Dumbledore locks the door to the hospital wing. We've got to rescue Sirius and get back into the ward before anybody realizes we're missing†¦.† They waited, watching the moving clouds reflected in the lake, while the bush next to them whispered in the breeze. Buckbeak, bored, was ferreting for worms again. â€Å"Do you reckon he's up there yet?† said Harry, checking his watch. He looked up at the castle and began counting the windows to the right of the West Tower. â€Å"Look!† Hermione whispered. â€Å"Who's that? Someone's coming back out of the castle!† Harry stared through the darkness. The man was hurrying across the grounds, toward one of the entrances. Something shiny glinted in his belt. â€Å"Macnair!† said Harry. â€Å"The executioner! He's gone to get the Dementors! This is it, Hermione –â€Å" Hermione put her hands on Buckbeak's back and Harry gave her a leg up. Then he placed his foot on one of the lower branches of the bush and climbed up in front of her. He pulled Buckbeak's rope back over his neck and tied it to the other side of his collar like reins. â€Å"Ready?† he whispered to Hermione. â€Å"You'd better hold on to me –â€Å" He nudged Buckbeak's sides with his heels. Buckbeak soared straight into the dark air. Harry gripped his flanks with his knees, feeling the great wings rising powerfully beneath them. Hermione was holding Harry very tight around the waist; he could hear her muttering, â€Å"Oh, no — I don't like this oh, I really don't like this –â€Å" Harry urged Buckbeak forward. They were gliding quietly toward the upper floors of the castle†¦. Harry pulled hard on the left-hand side of the rope, and Buckbeak turned. Harry was trying to count the windows flashing past — â€Å"Whoa!† he said, pulling backward as hard as he could. Buckbeak slowed down and they found themselves at a stop, unless you counted the fact that they kept rising up and down several feet as the Hippogriff beat his wings to remain airborne. â€Å"He's there!† Harry said, spotting Sirius as they rose up beside the window. He reached out, and as Buckbeak's wings fell, was able to tap sharply on the glass. Black looked up. Harry saw his jaw drop. He leapt from his chair, hurried to the window and tried to open it, but it was locked. â€Å"Stand back!† Hermione called to him, and she took out her wand, still gripping the back of Harry's robes with her left hand. â€Å"Alohomora!† The window sprang open. â€Å"How — how –?† said Black weakly, staring at the Hippogriff. â€Å"Get on — there's not much time,† said Harry, gripping Buckbeak firmly on either side of his sleek neck to hold him steady. â€Å"You've got to get out of here -the Dementors are coming — Macnair's gone to get them.† Black placed a hand on either side of the window frame and heaved his head and shoulders out of it. It was very lucky he was so thin. In seconds, he had managed to fling one leg over Buckbeak's back and pull himself onto the Hippogriff behind Hermione. â€Å"Okay, Buckbeak, up!† said Harry, shaking the rope. â€Å"Up to the tower — come on.† The Hippogriff gave one sweep of its mighty wings and they were soaring upward again, high as the top of the West Tower. Buckbeak landed with a clatter on the battlements, and Harry and Hermione slid off him at once. â€Å"Sirius, you'd better go, quick,† Harry panted. â€Å"They'll reach Flitwick's office any moment, they'll find out you're gone.† Buckbeak pawed the ground, tossing his sharp head. â€Å"What happened to the other boy? Ron?† croaked Sirius. â€Å"He's going to be okay. He's still out of it, but Madam Pomfrey says she'll be able to make him better. Quick — go –â€Å" But Black was still staring down at Harry. â€Å"How can I ever thank –â€Å" â€Å"GO!† Harry and Hermione shouted together. Black wheeled Buckbeak around, facing the open sky. â€Å"We'll see each other again,† he said. â€Å"You are — truly your father's son, Harry†¦Ã¢â‚¬  He squeezed Buckbeak's sides with his heels. Harry and Hermione jumped back as the enormous wings rose once more†¦ The Hippogriff took off into the air†¦ He and his rider became smaller and smaller as Harry gazed after them†¦ then a cloud drifted across the moon†¦. They were gone.

Regionalism: Faulkner “A Rose for Emily” Essay

Emily Grierson- A mysterious woman who had secrets kept and a puzzlement to the community around her. †¢Colonel Sartoris- The man who reassured her that she would not need to pay taxes or anything. †¢Tobe- A loyal servant that fed and took care of Miss Emily as well as her secrets of which he never told anyone. †¢Judge Stevens- He acquired new solutions to the community complaints of and about Miss Emily I suppose as respect for the elderly woman. †¢Homer Barron- A man that came into Miss Emily’s life who after a while was not seen again until the day Miss Emily died and was found in a bedroom upstairs decaying. Part II: â€Å"A Rose For Emily† Questions 1.What metaphor is used to describe Miss Emily in the first paragraph? In the first paragraph they described Miss Emily as a Fallen Monument. 2.How is the house personified in the second paragraph of the story? The house is personified as a beautiful house at first with scrolled balconies, cupolas, spires and beautiful flowers. Then towards the ending of the paragraph the house is withered with time and left un developed while as the houses around hers develop and acquire a new style. Miss Emily’s house was described as an eyesore. 3.What had Colonel Sartosis done for Miss Emily in 1894? Colonel Sartosis remitted Miss Emily’s taxes with a dispensation dating from the death of her father and on to perpetuity. 4.What did the next generation of town leaders do on the first year? The alderman issued Miss Emily a notice to pay her taxes and when that hadn’t been claimed they issued another and when the same happened they decided to send people to her house to have a discussion on the issue. 5.How does Faulkner describe Miss Emily in the Sixth paragraph? In the sixth paragraph Miss Emily is described as a small, fat woman in black, with some jewelry and an old and weathered face. 6.At the beginning of Part II, how long had Emily’s father been dead? At the beginning of Part II in the story Miss Emily’s father had been dead for 2 years. 7.What are the neighbors complaining about? What does Judge Stevens say probably has caused it? Judge Stevens is being complained to by the community about the odor of Miss Emily’s home. The judge says that it is probably a dead snake or animal that her Negro servant has killed. 8.What did Miss Emily tell her visitors the day after her father’s death? After hearing of Miss Emily’s fathers death the community come to the house to share their condolences and when they do Miss Emily tells them that her father isn’t dead only to find her realization 3 days later. 9.Who began to date Miss Emily in Part III, and why was he in town? Homer Barron a young construction worker dated her the summer after her father’s death. Homer was in town to pave the sidewalks. 10.What did the townspeople think of Miss Emily and her new boyfriend? At first people were glad for Miss Emily but others thought that it wasn’t right because of Homer’s job. 11.What does Miss Emily do to make the townspeople think that she and her boyfriend have wed?Miss Emily went and bought a suit and nice dress shoes.The towns people thought that Miss Emily and Homer would marry but as time passed by a wedding wasn’t held and the ladies of the town thought that it was a bad example for the younger people so finally they sent the Bishop to Miss Emily’s home. When the Bishop returned from the home he did not speak of the interview. The Bishop’s wife wrote to Miss Emily’s family and when the ladies heard this they thought that Miss Emily had wed. 12.When was the last time the townspeople saw her boyfriend/husband? One evening a neighbor saw Homer welcomed in to Miss Emily’s home shortly after Miss Emily’s cousin’s left town. That was the last that the town saw of Homer Barron. 13.Why had the men sprinkled lime around her house in Part II? After some complaints to the judge of the odor of Miss Emily’ s house the men wanted to confront Miss Emily but the judge said it would be rude so he told them to sprinkle lime around the Home so that the odor would be gone. 14.There is a room upstairs no one has seen for over forty years. After Miss Emily’s funeral, the door to this room is broken down. What do the townspeople find there? After breaking down the door the towns people find the clothes that Miss Emily has bought shortly after Homer came in to her life and also Homer himself. 15.What happened to Homer Barron? Comment on the second pillow on the bed in the last paragraph while responding to this one. Homer Barron was found dead. I believed he was murdered with the poison that Miss Emily had bought from the druggist. A strand of Miss Emily’s hair was found on the pillow next to where he lay.