Alcoholism essays

Alcoholism essays Alcoholism is a chronic disease caused by a craving for alcohol. People suffering from this illness are referred to as alcoholics. They do not know how to control there drinking problem even when it causes a serious harm to their health and this could lead to medical disorders, marital difficulties, job loss, or automobile crashes. Alcoholism can be influenced by genetic, psychological and social factors. It cannot be cured, but many treatment options can help an alcoholic to avoid drinking and live a healthy and safe life. When a person consumes alcohol, the stomach and intestines rapidly absorb it. From there alcohol travels in the blood throughout the entire body, affecting nearly every tissue. High doses of alcohol depress the functions of the central nervous system, including the brain. The higher the alcohol level in the blood, the greater the impairment (Encarta). As the blood passes through the liver, enzymes break down alcohol into byproducts, which are eliminated after six to eight hours from the body. The alcohol remains in the body, producing intoxicating effects after the last drink is taken. Smaller quantities of alcohol reduce inhibitions and produce feeling of relaxation and happiness. Larger amounts of alcohol causes greater impairment to the brain and usually impairs sexual performance, particularly in males. When a person is intoxicated, theyre thinking and perception skills become blunt. Alcohol reduces anxiety, guilt, worries and often leaves a person feeling good about themselves (Goon 261). If drinking continues to happen it can lead to loss of physical control and possibly death. There are three types of effects of alcohol on various organs and tissues. These effects include the short, long and withdrawal symptoms. During the short-term effects the brain is mostly affected. The drinkers judgement and thinking becomes dulled. The perso...

17 Funny Halloween Quotes to Make You Laugh

17 Funny Halloween Quotes to Make You Laugh Halloween is a festival of chills and thrills. Gear up to meet people in bizarre costumes. Join the Halloween celebration with your own brand of freaky fetish. Bake skull cookies, mix dragon blood beverages, and eat eyeball scones. Surprise your guests with funny Halloween quotes scribbled on your wall as graffiti. You can make your Halloween party a memorable one with a touch of creativity. Halloween Around the World Halloween traditions vary by geography. If you are in Austria, you would probably leave some bread and water for the departed souls. Ireland, from where Halloween is said to originate, celebrates Halloween in spectacular style. Visit Dublin, Ireland, for the Halloween festival fiesta. In the U.S., Halloween is second only to Christmas. Children go door-to-door in spooky costumes, collecting treats from friendly neighbors. Adults have â€Å"Halloween nights† with themed parties and pumpkin pies. The Swedish like to celebrate Halloween by remembering their deceased friends and relatives. Honoring the dead by lighting candles by their graves is the Swedish Halloween tradition. Other countries like Russia, Mexico, Sweden, Germany, Japan, and Madagascar also celebrate Halloween in their own unique way. Whatever country you are in, make sure that you have a fun-filled Halloween night. Here are some funny Halloween quotes to get you into the spooky mood! Halloween Quotations Rita Rudner Halloween was confusing. All my life my parents said, Never take candy from strangers. And then they dressed me up and said, Go beg for it. I didn’t know what to do! I’d knock on people’s doors and go, Trick or treat. No, thank you. Fernando Pessoa Look, theres no metaphysics on earth like chocolates. Jean Baudrillard There is nothing funny about Halloween. This sarcastic festival reflects, rather, an infernal demand for revenge by children on the adult world. Rodney Dangerfield On Halloween, the parents sent their kids out looking like me. Richard Harris Barham Ghosts, like ladies, never speak till spoke to. Lloyd Douglas If a man harbors any sort of fear, it makes him landlord to a ghost. Miguel de Cervantes Fear has many eyes and can see things underground. Anonymous Ill bet living in a nudist colony takes all the fun out of Halloween. Nina Willis Walter The witches flyAcross the sky,The owls go, Who? Who? Who?The black cats yowlAnd green ghosts howl,Scary Halloween to you! Scottish Saying From ghoulies and ghosties and long-leggedy beasties and things that go bump in the night, Good Lord, deliver us! Steve Almond Nothing on Earth so beautiful as the final haul on Halloween night. Dee Snider Halloween is huge in my house and we really get into the spirits of things. Conan OBrien This Halloween the most popular mask is the Arnold Schwarzenegger mask. And the best part? With a mouth full of candy you will sound just like him. George Carlin There are nights when the wolves are silent and only the moon howls. Henry David Thoreau I would rather sit on a pumpkin and have it all to myself than be crowded on a velvet cushion. Mark Twain Everyone is a moon and has a dark side, which he never shows to anybody. Drew Carey I see my face in the mirror and go, Im a Halloween costume? Thats what they think of me? Source Morrison, Patt. Halloween is turning into a worldwide holiday. Why do so many people love to be scared? Los Angeles Times, October 31, 2018.

Ethical and Socially Responsive Business Essay Example | Topics and Well Written Essays - 750 words

Ethical and Socially Responsive Business - Essay Example This enables the restaurant to maintain high social and ethical standards in the society where it carries out its business. This implies the restaurant must be accountable for the negative effects of its operations on the government, suppliers, consumers, the civil rights groups and churches within a given jurisdiction. The restaurant is committed to safeguarding the environment from pollution from its own business. Moreover, it has embarked on the creation of good social relations with the society around its location in order to gain an understanding and gather information on the impact of the restaurant’s activities on the social environment (Chipotle Mexican Grill, Inc, 2). This restaurant is known to incorporate its stakeholders in the decision making process. This gives the community a chance to give a credible contribution with regard to the restaurant’s social responsibility. Social responsibility has enabled the restaurant to have strong influence within its loc ality given its participation in provision of externalities to the society. The restaurant code of conduct with regard to insider information applies to its business. The activities of this restaurant are many given its competitive advantage as a food industry leader. The entity receives credit from various customers but insider information must be kept the restaurant’s secret all the time. This enables the restaurant to access financial aid. Moreover, this serves as a sales and marketing technique because a financially stable enterprise is considered favorable when its success strategies are not exposed. According to Carroll and Ann, compliance to the safeguarding of insider information requirement contributes in the market liberalization hence enabling the consumers to make informed decisions (38). It is involved in the handling of consumer issues. This has made many consumers and other stakeholders to identify with the restaurant hence outstanding performance in the sellin g of its foodstuffs. Moreover, the code of conduct pertaining to harassment and sexual abuse of women ensures that women are protected and appreciated hence female customers of Chipotle restaurant have increased. This has increased the overall returns of the restaurant (Chipotle Mexican Grill, Inc, 2). The anti-discrimination regulation anchored in the code is also relevant in the running of this restaurant. This offers the customers protection against any intimidation based on race or tribe hence increasing the restaurant revenue. Hancock asserts that anti-discrimination gives the stakeholders of the restaurant concerned an assurance of the safety of their resources (67). The requirement of the code pertaining to sexual harassment also boosts the customer relationship within the restaurant given that women feel more secure and accommodated. This boosts the revenue because of the influx of many ladies into the restaurant. Integrity is another aspect which is vital in the restaurant. This aspect has a high correlation to thrive in the restaurant business. The workforce must act in utmost good faith at all times in order to realize favorable performance results at the end of the financial period of the restaurant. The management also must exhibit transparency and good stewardship of the economic resources entrusted to them by the stakeholders. According to Carroll and Ann compliance to this requirement results in productivity and

Global Marketing Assignment Example | Topics and Well Written Essays - 1500 words

Global Marketing - Assignment Example Culture is recognised to be an essential factor which imposes considerable impact upon the entire performance of the organisations (Lee & Carter, 2009; Bardhan & et. al., 2006). Culture is recognised to be an essential element on the basis of which the different magnitudes of human behaviour are determined. Culture comprises different elements that include belief, morals, knowledge, art and customs among others. These are various elements that depict the culture of a particular society. Moreover, culture is based on different macro environmental factors including social, economic, political and religious beliefs. In this regard, people of a society develop cultural values, behaviour and beliefs on the basis of the aforementioned factors (Soares & et. al., 2007). Culture is based on various concepts such as attitudes, values and ideas through which people perform various actions in a society. The cultural values and beliefs are usually shared amid the people of a society according to which individuals develop their ethnic values and purchasing behaviour (Belshek, 2004). Global marketing is often regarded as an appropriate strategy which adopt by business originations with the intention of conducting diverse operational functions effectively on a global context. Presently, there are various factors that can be viewed to be responsible for business organisations to execute their respective operations successfully in overseas business market segments. In this regard, these factors comprise minimised or unrestricted trade barriers, multilateral agreements in relation to trade activities and prevalence of common business market conditions. Culture is often recognised to play an imperative role in supporting the organisations towards making effective decisions. Business organisations, in order to execute their respective operational functions or activities efficiently in the worldwide market segments, are required to possess adequate knowledge in relation to fundament al and cultural values. In this respect, organisations having adequate knowledge in relation to cultural values will be able to operate their business operations with greater efficiency. It is worth mentioning that organisations, with appropriate understanding and knowledge about cultural values and beliefs will be facilitated with the opportunity of formulating along with executing effective marketing strategies towards the accomplishment of predetermined business targets (Vogt, 2013; Kotabe & Hesen, 2010; Gillespie & et. al., 2010; Lee & Carter, 2009). Culture should be considered in global marketing as the present business market scenario or the market conditions is recognised to be changing frequently due to several factors. The foremost factors for the change in the market conditions can be apparently observed as the development of new business markets and the preferences of global consumers that are likely to alter in relation to demographics along with expectations. It has be en viewed that the global marketing activities have changed due to the dissimilar cultural values along with beliefs that holds by the worldwide customers (Keegan, 2011; Lee & Carter, 2009; Passaris, 2006; Picard, 2004). Thus, it can be affirmed that the market conditions of global market are often predicted to remain much dependent

The Role Of Patriarchy In Female Characters English Literature Essay

The Role Of Patriarchy In Female Characters English Literature Essay A similarity exists in both the authors writing which is in disagreement to the dominant culture and the standard societal ideas of their time. This is shown by the two modern works which I will be analyzing; Isabel Allendes The House of Spirits and A Dolls House by Henrik Ibsen which develop the conflict of gender roles in their works. Both authors use in my opinion an authentic female character(s) and that by the end despite the male domination are able to rebel against societal norm only to be viewed as the protagonist of the play or novel. Allendes novel recounts the story of Estaban Trueba, his wife and children. With the story spanning three generations is off of political corruption, feminine oppression and the movement from old to new. Allende examines and reveals the internal conflict of turmoil, oppression and torture of the female characters in particular. Women have generally been considered silent figures, submissive to the patriarchal powers that govern their society. Neither Allende nor Ibsen attempt to reproduce the culturally ordained silence; instead, both re inscribe female silence as a means for female empowerment which leads to their eventual freedom. The period, in which the novel was written, women were confined by traditional gender roles, with most women performing the work traditionally designated for women: marriage and family, or, if employed outside the home, teaching. Isabel Allende strongly believes that empowerment leads to the strength and freedom of a woman; and these views were shared by Henrik Ibsen as this concept echoes throughout their works. From the early sections of the novel, both Nivea and Clara are involved in the suffragette movement and this was not granted to women until 1949. Their participation in the Suffragette group would have been considered socially unacceptable as the right of voting would only be given to the male members, establishing from the very start that Chile is a patriarchal society. Allende tries to establish that empowerment comes through force of conviction, and uses Nivea as an example to show this. We see that women such as Nivea who have acted against societys norm by fighting for political rights go further than those women who accept their traditional role of subservience and remain staunch in their conservatism. They are shown to finish their days alone and mostly forgotten as is seen with Ferula and Nana. The portrayal of Nivea and her funeral which was attended by many is in direct contrast with that of Nana and Ferula. It is quoted that with Nana None of the many children she had raised with so much love attended her funeral.(P 17). Nana is said to have: been born to cradle other peoples children, wear their hand-me-down clothing, eat their left overs, live on borrowed happiness and grief, grow old beneath other peoples roofs die one day in her miserable little room in the far courtyard in a bed that did not belong to her, and be buried in a common grave in a public cemetery.(P 57) Allende uses Nana to sadly reflect on the women who meekly accept their defined role in society of the time. Allende does not only express empowerment through conviction but also through commercial enterprise as seen with Transito Soto. At the start of the novel Esteban lends her 50 pesos allowing her to seek out a career, but by the end of the novel the tide turns and the arrogant sexist Esteban comes to her to beg a favor. Again, by women taking chances and setting goals and dreams which society restricts them from having end up being successful. Allende shoots her views to the reader through her writing by using these female characters to represent Allendes own brand of feminism. Allende establishes early in her novel that when the family was warned of Claras nine-year silence and psychokinetic skills, Trueba considers the prolonged silencea virtue (88). Such a reading of muted female voice is constant in Latin American culture. However we see that the protagonists of the novel are all women who work in different and subtle ways to assert their rights breaking this silence in an assertive manner to get what they want and need in life. Still, . Clara, Blanca, and Alba remain the focus of the story, while Esteban, Pedro Tercero, and Miguel though the males and automatically assumed superiors of society enter the story only because they are the men those women love or marry. Though they are women and regarded as inferiors to society The House of the Spirits shows that this does not mean that men accomplish things and change things while women do not. On the contrary, the women in The House of the Spirits have a more long-lasting effect and create more drastic changes than any of the men do. This has been done by disapproved behavior such as Nivea and her movement or Transito and the prostitute business while the men lead revolutions that topple governments, those revolutions are themselves quickly toppled. Henrik Ibsens play A Dolls House chronicles the attempts of one woman to find freedom in a patriarchal society. The feminist issues in A Dolls House seem almost contemporary, although it was written more than a century ago. Like Allende Dolls House challenges these patriarchal views and shows the audience a fresh perspective on a womans life. Many women could relate to Noras situation and like Nora, they felt trapped in a dolls house by their husbands and their fathers; however, they believed that the rules of society prevented them from stepping out of the shadows of men. Through this play, Ibsen stresses the importance of womens individuality and like Allende Ibsen uses his characters of the play help to support his strong opinions. Noras initial characteristics are that of a bubbly, child-like wife who is strictly dependent on her husband. This subordinate role from which Nora progresses emphasizes the need for change in societys view of women. For Nora, her inferior, doll-like nature is a facade for a deeper passion for individuality that begins to surface during the play and eventually fully emerges in the ending. An example of this deep yearning for independence is shown when Nora tells her friend, Kristina Linde about earning her own money by doing copying. Nora explains, It was tremendous fun sitting [in her room] working and earning money she says; It was almost like being a man (162) Such comments reveal Ibsens presentation of women to be positive; Nora enjoyed working because it empowered her, and whilst accepting the situations of the time, Ibsen portrays women as active, and struggling in an unequal system. The sheltered, feeble wife proves to be a resourceful masculine life-provider, in literally sav ing Torvalds life. In many instances Nora deceives her husband, even though Nora seeks to be independent, she uses her role of obedience to her advantage and does this by deceiving Torvald into thinking that she can do little on her own, she ensures that he never suspects her of forging her fathers name to borrow 800 cronen from Krogstad in order to save Torvalds life. When Krogstad threatens to expose the truth, Nora must use her craftiness to distract Torvald and sway him into letting Krogstad keep his job. She soon comes to realize the she must take the path that is right for her and no one else. She discovers that Torvald is not the man she thought him to be and that he knows nothing of who she really is. In her sudden awareness, she says to Torvald, you dont understand me. And Ive never understood you until tonight. They have lived as strangers to each other. Noras progression from a submissive housewife to an opinionated, independent woman represents the future progression of women in society. Torvalds role illustrates societys inaccurate perception of women. He sees her only as a one-sided character with little or no opinions of her own. As Nora comments, I came to have tastes as yoursor I pretended to. Noras father also contributes to her oppression. He used to tell [Nora] his opinion about everything, and so [she] had the same opinion. Torvald only continues the behavior that she had come to expect from her father. These male figures in Noras life not only limit her beliefs and actions, but also limit her happiness. Like Allende Ibsen uses Torvalds character, and discards the public view of man to expose a more realistic depiction of male superiority. Ibsen attempts in criminalizing the male protagonist; Torvald, his behavior becomes increasingly more erratic near the plays end, affirming an idealistic self-importance, believing that she has become wife and child to him. Moreover, Torvald exhibits considerable emotional disparity which one may stereotypically associate with women; he becomes angry at Nora for her deceptive act, yet once he appears to be in the clear, he deems his outburst to be a moment of anger. Just as Torvald undergoes an apparent transformation, so does Nora; she proclaims that her leaving Torvald and the children is necessary for her, a notion that is typically assumed by males (to consider ones interests first). To portray Nora as such may not be entirely positive, but it is certainly a contemporary, liberal representation. Overall, we see that Nora is now free from her obligations, and rejects Torvalds further assistance. Moreover, Nora the doll (one whose disposition never changes) now leaves her house, and is free to seek a fuller life as a human being, no longer a malleable doll under the control of her master. No women at the time would have dared to do what Nora has, and just like the characters in Allendes novel it is only by daring or committing actions socially unacceptable like Nora did or even as Nivea displayed that these women can truly feel empowered which leads to their freedom or gain of what they needed. Again Allende and Ibsen reverberates their thoughts through the characters and portray the males as the antagonists of the play which leads to the audience or readers to understand and sympathize with their views that their needs to be a change in the way society views women.

The Predatory Organism: Bdellovibrio

Identifying the genes and the proteins to be expressed by this organism during pore formation in the host cell will be of significance to bacteriology. The possible use of this information can be seen in the study done by Kadouri and O’toole. It was established in this report that B. bacteriovorus can be used as a control agent against certain biofilm communities. It was observed that biofilm populations of Escherichia coli and Pseudomonas fluorescens were significantly reduced when exposed to B. bacteriovorus.Since biofilm formation is unnecessary and damaging in some instances, inhibition of its formation can be done through natural elimination of the biofilm organisms, and that is through the inoculation of B. bacteriovorus. Another possible application of this predatory organism is through its use as an anti-microbial therapeutic agent. It had been suggested by Hobley et al. that the introduction of this organism to a microbial-infected wound can lead to the reduction of p athogenic organisms in the site of inoculation.In this way, the use of antibiotics can be avoided and possible resistance of disease-causing organisms to the applied drug can be avoided. If the genes responsible for pore formation in the host cell will be identified, the organism can be manipulated to increase the genes’ expression. These can then be enhanced and controlled in accordance to the desired amount and time of expression. II. Aims This paper aims to propose a method on how to determine the necessary and significant enzymes involved in pore formation of the host cell by Bdellovibrio bacteriovorus H100.The specific aims are as follows: 1. Identify the genes of B. bacteriovorus H100 specifically expressed during pore formation in the host cell during the predatory life cycle phase of the organism using time course Microarray analysis. 2. Identify specific hydrolytic enzymes expressed by the identified genes of B. bacteriovorus H100 involved in pore formation on host c ell membrane. III. Background The discovery of Bdellovibrio by Stolp and Starr in 1962 opened the scientific world to the dawn of a new organism- a bacterium which preys on its own kind (12).This bacterium is a curved rod, Gram negative organism classified under the group of delta-proteobacteria. It is a motile bacterium; monotrichously flagellated; obligately aerobic; small in size (0. 3 ? m in width and 1-2 ? m in length); and is obligately predatory. Its genome consists of 3. 85 mega base pairs. Most of these genes encode for enzymes involved in hydrolysis and flagella involved in host sensing (3). The best known species of Bdellovibrio is B. bacteriovorus, observed to prey on other Gram negative organisms such as Salmonella, Escherichia coli, Sphaerotilus natans and Pseudomonas fluorescens (1).The distribution of Bdellovibrio bacteriovorus is observed in diverse environments; therefore, it is considered to be an ubiquitous bacterium. It was associated in dry environment such as the soil; in wet locations such as brackish water, sewage, fresh water, pooled reservoirs and sea water; and in unique microbial niches such as biofilms (3). The life cycle of this bacterium consists of two stages. The first stage is the so-called free-swimming attack phase and the second stage is known as the intraperiplasmic replication phase. Figure 2 shows a graphical representation of these two stages.The life cycle of B. bacteriovorus takes about three hours to complete. For the initiation of the attack, the monotrichous flagellum is an important facet to consider. Movement is essential in finding the suitable host in the environment. The predatory organism moves towards a region with a high prey concentration. This process is meditated by chemotaxis. It can be seen from the illustration that a critical stage in the predation of B. bacteriovorus is the ability of the organism to penetrate its host bacterium. Upon contact with another Gram negative, B.bacteriovorus then forms a pore in the cell membrane of its host. Initial entrance of B. bacteriovorus is followed by the organism’s penetration of the periplasm. Bdellovibrio bacteriovorus then occupies the periplasmic space of the host cell (3). Without this phase, the other steps in the whole life cycle of the organism cannot be ensured to occur. Figure 3 and 4 shows an electronmicrograph of B. bacteriovorus attaching to a host cell. A specific strain of the bacterium, B. bacteriovorus HD100, was studied by Rendulic, et al. This strain of Bdellovibrio was found to have an unusually large genome.Though this bacterium preys on other Gram negative organisms, its genetic make-up did not comprise of any gene from its host. Furthermore, it was elucidated that the genes present in B. bacteriovorus HD100 are made up of gene families coding for enzymes such as hydrolases and transporters, important in the penetration and killing of the host. These genes also code for enzymes needed for uptake of complex mol ecules (6). One hurdle in studying the molecular characteristics of this organism is its host dependent nature. Without a suitable host, growth cannot be ensured, thus, elucidation of its genetic make-up may be difficult to achieve.Further studies using this bacterium revealed that Bdellovibrio can generate mutant cells that do not require host cells for growth and are therefore known as host independent (HI) strains. Despite this, they were able to retain the ability to grow on prey and hence are termed as facultative predators. For gene manipulation techniques, HI strains are usually used (8). Despite the fact that the complete genome of the organism was already sequenced, the specific genes coding for the needed enzymes to form pores in the host cell were still unidentified.With this lack of information, this study is formulated and designed. IV. Research Design and Methodology Culturing of B. bacteriovorus HD100 on prey dependent and prey independent set-ups: Predatory (HD) cult ures of B. bacteriovorus HD100 will be grown on E. coli in Ca2_-HEPES buffer at 30Â °C, with shaking at 200 rpm (8). Escherichia coli ML35 and E. coli W7-M5 (10) will be used as the prey throughout the experiments. Escherichia coli ML35 will be cultured in nutrient broth (Difco Laboratories), and E. coli W7-M5, a lysine and DAP auxotroph, will be cultured in nutrient broth supplemented with 0.2 mM lysine and 0. 1 mM DAP at 37Â °C with shaking at 200 rpm. Prey-independent HI strains will be plated on rich peptone-yeast extract (PY) medium (8). Synchronous cultures: Synchronous cultures will be used for performing various experiments as described below. Briefly, fresh bdellovibrios will be added to prey cells in HM buffer (3 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)-1 mM CaCl. LQ. One mM of MgCl2 will be adjusted to pH 7. 6 using NaOH (10). The organisms will be grown until a final concentration of 1010 bdellovibrios per ml and 5 x 109 E.coli per ml is reached. For proper aeration, volumes will be kept to ? 20% of the flask’s volume and incubated at 30Â °C with shaking at 400 rpm. Synchronous cultures will be examined at intervals for attachment and penetration with a Nikon model L-Ke microscope (Nippon Kogaku Inc. ) equipped with phase-contrast optics and a Nikon model AF camera. Time course Microarray analysis. Time course Microarray analysis will be performed to identify the genes to be expressed during the entry phase, specifically during pore formation on the host cell membrane of B.bacterovorus H100. Microarray slides of B. bacteriovorus H100 will be ordered from Advanced Throughput, Inc Services. Total cellular RNA will be extracted from B. bacteriovorus H100 cells at entry phase using the RNeasy mid kit (Qiagen). The RNA of the organism will also be extracted during the other stages of infection. This will serve as a reference for comparison of the genes expressed and not expressed at the desired stage. Complementary DNA sy nthesis, fragmentation, labeling, hybridization, staining and washing will be performed according to the Affymetrix B.bacteriovorus H100 GeneChip array expression analysis protocol (Affymetrix). Briefly, cDNA will be synthesized from RNA using Superscript II (Invitrogen) according to the manufacturer’s instructions. RNA will be removed by alkaline treatment and subsequent neutralization. Complementary DNA will be purified with QIAquick PCR purification columns (Qiagen). Purified cDNA will be fragmented by DNase I (Amersham) at 37Â °C for 10 min followed by end labeling with biotinddUTP, using an Enzo BioArray terminal labeling kit (Affymetrix), at 37Â °C for 60 min.Hybridization will be performed in an Affymetrix GeneChip hybridization Oven 640. Washing and staining will be performed using an Affymetrix Fluidics Station 400. Arrays will be scanned with an Agilent GeneArray Scanner G2500A. GeneChip scans will be initially analyzed using the Affymetrix Microarray Suite 5. 1 s oftware, from which PivotData tables will be exported. Raw data from the PivotData Tables will be analyzed in GeneSpring software version 6 (Silicon Genetics), using the parameters suggested by Silicon Genetics for analysis of Affymetrix Microarrays. Real-time PCR:Real-time PCR using the Applied Biosystems 7500 Real-time PCR system will be performed to confirm microarray results. RNA will be extracted from B. bacteriovorus H100 at initial phases of predatory life cycle up to entry phase as described above. RNA will be reverse transcribed into cDNA and simultaneously labelled using the iScript One-step RT-PCR kit with SYBR Green (Biorad). RT-PCR reactions will also be performed to amplify cDNA of housekeeping genes (identified from micro array studies) for normalization of fluorescence values. Identifying the specific hydrolytic enzymes of B.bacteriovorus which are involved in pore formation on host cell membrane. Many experiments showed that B. bacteriovorus H100 releases hydrolytic enzymes during predatory life cycle. According to Thomashow and Ritterberg, glycanases and lipopolysaccharideases are required for pore formation in the prey’s peptidoglycan and LPS layers respectively. The glycanase and/or peptidase could be responsible for weakening the peptidoglycan layer of the prey and thereby responsible for permitting conversion of the substrate cell to a spherical shape (10).Tudor et al. proposed another model for penetration. According to them peptidase is responsible for pore formation but not glycanase (11). Specific enzymes involved in pore formation are not known. The genes identified from the time course micro array technique will be mutated as described previously using suicide vector pSSK10. Resulting mutants will be complemented by using vector pMMB206 (8). Mutants will be analysed for the specific enzymes (using 2D-gel electrophoresis) and their actions on host cell i.e, as a glycanase, LPSase or peptidase will be observed by radio labellin g experiments (10). Wild-type B. bacteriovorus H100 and complemented strains will be used as controls. Radio labeling experiments: Escherichia. coli W7-M5, auxotroph for lysine and DAP and cannot metabolize glucosamine, will be radiolabelled as described previously (9,10). Peptide portion of E. coli W7-M5 peptidoglycan will be labelled with [3H] DAP and the lipopolysaccharides and glycan portions of the peptidoglycan will be labeled with [3H]glucosamine.Various mutants and wild-type strains will be tested for predation using this radiolabelled strain. Solubilisation of glucosamine and DAP from labelled prey peptidoglycan will be measured as described previously (11). Briefly, samples taken at intervals will be precipitated with an equal volume of cold 10% trichloroacetic acid for 30 min followed by centrifugation. Resulting supernatants will be assayed for soluble radioactivity in a scintillation counter (Rackbeta II). Two-dimensional gel electrophoresis: The hydrolytic enzymes rele ased by B.bacteriovorus H100 during its predatory life cycle will be analyzed by performing two-dimensional gel electrophoresis. Sample preparation for 2D-gel electrophoresis: Escherichia coli ML35 cells will be challenged with B. bacteriovorus H100 wild-type as well as the mutant strain. Culture fluid will be drawn from synchronous cultures during attachment and entry phases of B. bacteriovorus H100. Culture fluid will be centrifuged to discard any cell debris. Proteins in the supernatant will be precipitated using cold acetone. The precipitated proteins will be separated by centrifugation.The precipitated pellet will be air dried and will be dissolved in rehydration solution (8M urea, 2% CHAPS {3-[3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate}, 18 mM DTT, 0. 5% IPG buffer pH range 4-7; Amersham Biosciences), plus a trace of bromophenol blue. Sample protein concentrations will be determined using the BCA protein assay (Pierce). Resulting protein pellet will be subjected to 2D-gel electrophoresis. 2D-gel electrophoresis: Two-dimensional (2-D) gel electrophoresis will be performed according to the method of O’Farrell.Proteins present in the pellet will be resolved on two-dimensional gels using the products and protocols of Amersham Pharmacia Biotech. In the first-dimension, proteins will be resolved by isoelectric focusing on a precast Immobiline DryStrip with a linear pH gradient. This will be followed by electrophoresis using sodium dodecyl sulfate polyacrylamide gel on 12. 5% acrylamide gel. For analytical 2-D gel electrophoresis, 100 ? g of sample protein will be applied to the gels and the proteins will be stained with Pharmacia Biotech silver stain kit.For preparative two-dimensional protein gel electrophoresis, 500 ? g of the sample protein will be loaded on the gels and proteins will be visualized using Coomassie blue R-350 (Phast Gel BlueR; Amersham Pharmacia Biotech). Spot analysis will be carried out using PDQuest Image Analysis softw are (BioRad). Spots absent from the mutant sample gel will be manually excised from the wild-type B. bacteriovorus H100 sample gel for identification. Mass spectrometry and protein identification: Excised protein spots of interest will be destained, reduced, carboxymethylated, and digested with trypsin in situ.This will be done overnight with a temperature of 37Â °C as described (8). Gel digests will be centrifuged, and an aliquot of the supernatant will be taken for analysis using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Lists of peptide masses will be generated and searched against the NCBInr database using the Mascot protein identification system (Matrix Science; http://www. matrixscience. com). Sequences of proteins identified by Mascot will be analyzed for a predicted signal sequence using SignalP 3. 0 (http://www. cbs. dtu.dk/services/SignalP/). V. Conclusion The identification of genes responsible for the expression of hydr olytic enzymes of Bdellovibrio bacteriovorus is of significant importance. Since the organism preys on other bacteria, its special attributes must be thoroughly studied and appreciated for possible useful applications. As mentioned earlier, this predatory organism can be used as a control agent against biofilm, as well as pathogenic organisms. Through the use of natural agents to control destructive and disease-causing bacteria, the use of chemicals and antibiotics can be lessened or avoided.Since the rapid evolution of organisms towards resistance and tolerance to anti-microbial substances is becoming more widespread, an alternative approach to these problems can be useful in the world of bacteriology. Thus, the identification of these genes and hydrolytic enzymes is of utmost importance. The elucidation of this information can be used as a primary defence against harmful microbes if this knowledge will be taken advantage. A more effective predatory role can be performed by Bdellov ibrio bacteriovorus if the genes and the hydrolytic enzymes of the organism will be expressed efficiently and constitutively.

My Philosophy of Education Summary

In most of today’s classrooms we the students are seated in rows of desks. We are forced to sit and listen to the teacher, be it in biology, math, English, and memorize line after line of information. We are â€Å"receptacles† to be â€Å"filled† by the teachers. We have attained â€Å"knowledge† as an absolute if we complete the course with a %100. As society is seeing today, students being given the stamp of approval aren’t really ready to enter today’s world. If you’re going to school to be a biologist, it isn’t just enough to know that water enters and exits the cell through a process called osmosis. You need to know why! You need to be able to see all of this information on a much broader scope than it has been presented to you. Students need to be taught to absorb this knowledge rather than to memorize it. The main problem of today’s educational system is that it most of the times does not teach how to think critica lly. I propose a system of education in which teachers are not just depositing information in students, in which students are not just passive observers in an active world and in which knowledge is not just an absolute. The first thing that needs to be done to achieve my system (philosophy) of education is to change the way teachers are teaching all together. How many times do students talking amongst themselves outside the classroom about how good their teachers are. They go to class and interact with one teacher, and are taught everything from that one teacher’s perspective. In order to give the students a much broader perspective, two or maybe even three teachers, depending on the subject, should be placed in a class room. Each teacher would then present the information to the class. The students would then have a much broader spectrum of information to learn from. I am not implying that these teachers should tag-team forcing information down the student’s throat, but take different approaches in helping them absorb the information through problem posing. The teachers are not placed in this system to stand high up on their pedestals throwing assignments at the students, but are to b e working hand in hand with them. They would be learning from the students almost as much as they would be teaching them. In an educational environment such as this one, the students should be able to stand in the teachers place and teach the teachers and their fellow students as well. The students are active participants here. They should be learning as much from their peers as their teachers, if not more. This system could create a lot more â€Å"work† for the students, but not as much of the â€Å"busy work† we receive, and tire of, today. For example, students could be asked to do reflective writings, in which they would share what they have learned in the lessons. Student lead discussions would also take up a large portion of the time in these classroom environments. What better test of one’s knowledge than being put in a position where you are required to teach your fellow student? After each session both the student and the teach will go home feeling not stuffed with information, but a new view on some things that they would ponder as they head out into the rest of their dail y routines. The basis of this system isn’t what the teachers are teaching, or what the students are learning. It is how we perceive this thing we call knowledge. In this world knowledge isn’t something tangible. It is not viewed as an absolute. Knowledge is an ongoing process that will never have a climax or an end. It is something that will continue to grow and change through the timeline of our people. Knowledge is not what the students are learning, or what the teachers are teaching. It is the process of self-discovery. A teacher could tell a student that when you mix chemical A and chemical B they make chemical C. That isn’t the knowledge. The student can then actually mix chemical A and chemical B and watch them make chemical C. That isn’t knowledge. Knowledge is that individual’s learning of how those to things interact and why when they mix they form chemical C. Knowledge is the process of education through self-discovery. This system I have shown you would change the way education is viewed in this world. If we had teachers that actually â€Å"taught†, and students that questioned the answers they were given, we’d be much better off as a whole. If we keep going the way we’re going†¦ where are all the creative minds going to be coming from in the next few years? If students continue saying that A + B = C, what are we going to do when the world poses us more complex problems that require ingenuity, creativity and drive to solve? We just need to understand that knowledge is not just some gold star, or trophy each of us can achieve, but a method of learning and understanding that our society needs t