Living In Clip free essay sample

I dont know why folk-rock singer Ani Difranco hadnt thought of doing this sooner. A two CD set featuring live recordings of her remarkable, inspirational music from concerts around the country. Its terrific! A brilliant, wild, fun and evocative CD. A perfect amalgamation of almost all her best songs morphed into one long concert and whether you have experienced Ani live or not, youll love this CD! Many of Ani Difrancos fans, mainly young women who can relate to her poetic writing and raw sound, agree that her songs sound more vivid and life-like on stage than in a studio. Now you can have these 30 live songs on your personal stereo, experiencing every concert from Anis point of view. She tells her stories and sings her poems of fame, emotions, settings, boyfriends, girlfriends and making independent music. She also does an funky version of Amazing Grace, from her new 1996 album Dilate. We will write a custom essay sample on Living In Clip or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page Perhaps youre not familiar with Ani Difranco, whos sold more albums than Hootie and the Blowfish, all on her own record label Righteous Babe Records. Or maybe youre more into Jewel, Alanis Morissette or Fiona Apple. Hopefully this two-hour recording Living in Clip can outlive them. We all know theres good music out there, and Ani is it! .

Why Compromise was Necessary for US Government essays

Why Compromise was Necessary for US Government essays In the process of creating a new Federal government, inevitable conflicts made compromise absolutely necessary. At the Constitutional Convention in 1787, there were a number of differences between the delegates and the interests they represented. These delegates came from different backgrounds, and held different political views. One issue that was hotly debated was the argument of how many representatives each state should be allowed. The issue of including a Bill of Rights was also addressed. As a result, compromise was desperately needed. One major issue the delegates encountered was giving fair representation to both large and small states.. The large states favored Madisons Virginia Plan. According to the Virginia Plan, there would be a bicameral (2 houses) legislature with membership based on each states population. The voters would elect members of the lower house who would then elect members of the upper house. Both houses would vote for the countrys president and judges. This was clearly not fair to the smaller states, who would have less representation in this system. The Virginia Plan would give the largest states the majority of power. Instead, small states favored Patersons New Jersey Plan. The New Jersey Plan proposed a single-house congress in which each state had an equal vote. There were seven small states, meaning if they all voted together they would have the majority power. Once again, this system was not fair to the larger states. This conflict between the small and large states was so heated tha t it almost threatened to break up the Constitutional Convention. There was only one solution compromise. The compromise reached has come to be known as The Great Compromise. A delegate from Connecticut, Roger Sherman, proposed a two-house legislature, consisting of a Senate and a House of Representatives. Accordingly, Sherman had combined both the Virginia Pla...

The Logic and Evidental Problem of Evil Essay Example | Topics and Well Written Essays - 1000 words

The Logic and Evidental Problem of Evil - Essay Example All materials presented in this paper will come from class notes and a secondary source. Logical Problem of Evil Atheist J.L Mackie presented this argument where he argued that there is a contradiction between the existence of God and the existence of evil (Kelly 218). Mackie formulated his argument that there seems to be an evident contradiction between four main premises. One of them is that God is an omnipotent (Kelly 219). This premise translates that God is all-powerful, meaning that he is supposed to have the power of eliminating all evil that exists in this world (Meister 132). The second premise is that God is an omniscient. This second premise translates that God is all-knowing, meaning that he could have the ability to detect the occurrence and existence of evil in order to prevent or eliminate it. The third premise according to Mackie is that God is Omni-benevolent (Meister 132). This argument translates that God is all-good, meaning he would have the will to prevent all t he evils from happening in this world (Meister 132). The fourth premise indicates that evil and suffering exists in this world (Meister 132). This premise translates that there is evidence of moral and natural evil existence in this world. Reflecting on all of the above four premises Mackie concluded that God does not exist. The above four premises leads to a contradiction between the existence if evil and God. This derives to a point that God does not exist. For example, it is easy to reject the premise that God is omnipotent. This is because if God is not able to stop evil and most importantly know before it occurs in order to do something to stop it, then probably there is no God. There is also a possibility to reject the second premise; God is omniscient, by stating that if God is really all knowing, he could have the ability to stop evil. The above information indicates that no one is able to know what individuals can do even God himself. Moreover, this means that if God does e xist, evil occurs and surprises even him. It is also possible to reject the third premise indicating that God is Omni-benevolent. This is because if God is perfect good, he ought to have the ability of knowing when evil might occur and then have the power to stop it. However, it seems that God does not care about evil and possible he does not have the ability to stop it. Rejecting the first three premises translates into the fourth one, evil and suffering exists, and therefore giving a possible conclusion that there is no God. The Evidential Argument One of the well-known individuals who have firmly supported the evidential argument is William Rowe (Kelly 220). Through the evidential argument, Rowe and many other philosophers argue that much of the evils that people experiences in this world seem to be pointless. Rowe presented his arguments on four main statements. One of the statements indicated â€Å"E†, is an evil that no one established its justifier. However, many peopl e especially philosophers have been trying to find the justifier without any success. The second statement according to Rowe shows there is a high probability â€Å"E† has no justifier. The third statement indicates that if there are some evil that depict to have no justifier, then all the facts presented on theism about God depicts to be false (Meister 135). The fourth statement as presented by Rowe then concludes there is a high probability that theism is false. According the above statements

434Mod1Case Essay Example | Topics and Well Written Essays - 750 words

434Mod1Case - Essay Example Often cyber ethics education is based on one significant theory or combination of two theories. The two major theoretical approaches are teleological and deontological theories. Teleological theories states action’s consequences to be a measure of action’s good. On the other hand deontological theories consider action’s righteousness to be above goodness. Utilitarian foundation is a part of teleological theories. This foundation states that action of an individual should be able to maximize happiness and good for all who are affected by such actions. It highlights the factor that actions of a person should be based on probable consequences that may be resulted by the action before it is been executed. This theory claims that a person should think about each and every individual and not just person taking a decision. Deontological theory is an ethical study based on duties. It considers duty and fidelity toward principle to be most essential factors (Gold, 2010). This theory states that an action’s consequences are not that important in comparison to rightness of the action. As per this theory an action is said to be valid if actions that are taken by individuals do not lead to any form of contradiction. The basic concept of this theory is that individuals should work according to their intelligence and virtue; act justly, should tell truth and avoid any form of injury towards others. Both the theories have different elements which state the importance of cyber ethics. This form of education has gained its importance in the recent years where stealing music or video is considered to be equivalent to actual theft. Students are made aware about the different cyber crimes and its impact on others (Starr, 2011). Information technology in the present scenario has given liberty to individuals to access various data but also it has posed a threat to one’s privacy. In American schools

Discussion Questions Coursework Example | Topics and Well Written Essays - 250 words - 6

Discussion Questions - Coursework Example Saying anything at all is not doing something; one who says anything neither has control over what he says nor the consequences, most of which are normally negative. In light of this argument, saying anything means raising the eyebrows and counter-sayings or counter-actions along one’s path to a desired destination (Soccio 58). By advocating for inaction, it is fair to describe Asian philosophers as passive. Asia is technically a closed society where open reactions to issues are highly unlikely. In Buddha, the Middle Path is the cure for suffering (Soccio 59). The Path requires the faithful to avoid either extremes of life which would keep them in indulgence or denial. Successful individuals do not yearn for endless satisfaction, and they refrain from painful scenarios either. In all these, ego is always the trigger of all the negative stimuli in the brain, and thus it is the basis of denial and suffering. Tzu’s observation is accurate; Buddha’s approach to suffering ensures that adherents are always prepared to avoid suffering by staying away from extreme pleasures or overindulgence (Soccio 44). The approach can cure virtually all kinds of suffering because pain is often associated with both

Relationship between SMC1A and Chromosome Related Disease

Relationship between SMC1A and Chromosome Related Disease 1. Introduction[A1] The Structural Maintenance of Chromosomes 1 alpha (SMC1A) gene is located in Xp11.22-p11.21, consisting of 25 exons and 24 intron. SMC1A gene encoding a core subunit of the cohesin complex, which is essential to sister chromatid cohesion. SMC1, SMC3, SCC1 (also known as MDC1 and RAD21) and SCC3 (also known as SA2 and STAG2) subunits could interact with each other and form a ring-shaped cohesin complex [1-3]. As is known, central component of the cohesin and condensin complexes are required for conversion of interphase chromatin into mitotic-like condense chromosomes[4]. Structural Maintenance of Chromosomes (SMC) proteins are core component of the cohesin and condensin complex and essential for chromosome condensation during DNA replication and chromatid segregation of the genome in all organisms. They are also involved in checkpoint responses and epigenetic silencing of gene expression[5]. Cornelia de Lange syndrome is a dominantly developmental disorder with multisystem abnormalities including slow growth before and after birth, characteristic facial features, upper extremity defects, hirsutism, gastroesophageal dysfunction and cognitive retardaion. The incidence is as high as one in 10,000 to 30,000 newborns. Both sexes have the same phenotypic variability. To date, the three genes, NIPBL, SMC1A, and SMC3 involving in chromosome function, gene regulation and double-stranded DNA repair, could cause CdLS when mutated[6, 7]. Six in ten of the probands with CdLS have heterozygous mutations in NIPBL gene, whereas 5% have mutations in SMC1A and SMC3 genes [6, 8]. Eleven different SMC1A mutations in 14 unrelated patients have been reported. All patients had a mild to moderate CdLS phenotype [8-10]. In several decades, we focus on the relationship between SMC1A and chromosome related genetic disease. In recent years, we found that SMC1A may play a key role in tumorigenesis. Sun. M et al. determined that the effects of SMC1A knockdown on the cell cycle and apoptosis of lung adenocarcinoma cells. The results indicated that SMC1A is a novel oncogene, which modulates lung cancer cells in their proliferation and migration capabilities through arresting cell cycle at G0/G1 phase and promoting apoptosis [11]. The similar conclusion also was found in glioblastoma cells [12, 13]. However, SMC1A functions as a novel oncogene in human prostate cancer metastasis and progression has still not been reported. 2. Materials and Methods 2.1. Patient Samples All of the patient samples were obtained from the Urinary Surgery Department of Shanghai Changzheng Hospital, Shanghai, China. This study was approved by the Clinical Research Ethics Committee of Shanghai Changzheng Hospital, and the informed consents were acquired from all of the subjects. 2.2. Reagents and antibodies DMEM (cat no.12430-054), F12 (cat no. 21127022) and RPMI-1640(cat no. 11875-093) medium and fetal bovine serum (cat no. 10099-141) were purchased from GIO (Grand Island, NY). TRIzol Reagent was from Invitrogen (Carlsbad, CA, USA). Giemsa was from Chemicon International (Temecula, CA). M-MLV Reverse (cat no. M5301)Transcription was purchased from Promega (Madison, WI, USA). Oligo-dT(18) was synthesized by Sangon Biotech (Shanghai, China). Terraâ„ ¢ qPCR Direct SYBR ® Premix (cat no. 638318) was from Takara (Otsu, Japan). Anti-SMC1A antibody (cat no. SAB4300451) was from Sigma-Aldrich (Munich, Germany). Mouse anti-GAPDH (cat no. sc-32233), Goat anti-Mouse IgG (cat no. sc-32233) and goat anti-rabbit IgG (cat no. sc-2030) were from Santa Cruz Biotechnology (Texas, USA). All the other chemicals were of analytical grade. 2.3. Cell culture Human embryonic kidney cells 293T, Human prostate cancer cell lines PC-3, DU145, LNCap, and 22RV1 were purchased from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). 293T were cultrued in DMEM containing 10%FBS. PC-3 and DU145 cells were maintained in F-12 medium supplemented with 10% FBS, 100U/ml penicillin and 100ÃŽ ¼g/ml streptomycin. 22RV1 and LNCap cells were incubated with RPMI-1640 supplemented with 10% FBS, 100U/ml penicillin and 100ÃŽ ¼g/ml streptomycin. LNCap cells were maintained in Corning Corning ® CellBIND ® Surface cell culture flasks (Corning, cat no. #3289) for a better attachment efficiency. All cells were cultured in a humidified incubator at 37oC under 5% CO2 atmosphere and used for analysis during exponential phase of growth. 2.4. RNA interference The synthesized 21-bp oligonucleotides encoding SMC1A-specific shRNA held the sequence 5’-TAGGAGGTTCTTCTGAGTACA-3’. The sequence of the negative control shRNA oligonucleotides was 5’-TTCTCCGAACGTGTCACGT-3’. The oligos were annealed and ligated into pH-L vector (Hollylab, Shanghai, China) through NheI/PacI to generate pH-Lv-shSMC1A and pH-Lv-shCon. The resulting constructs were confirmed by sequencing. 2.5. Recombinant lentivirus Transduction PC-3 or DU145 cells were plated at 5Ãâ€"104 cell/well in 6-well plates. After 24 h of culture, lentivirus recombinant encoding shRNA against SMC1A was added at a multiplicity of infection (MOI) of 50 into F-12 basic medium. After 6h incubation, the cells were added with complete growth medium replacing the basic medium containing the lentivirus. Then, 5 days post-transfection, gene reporter (EGFP) expression was examined using fluorescent microscopy (Olympus, cat no. CKX41). 2.6. Quantitative real-time RT-PCR analysis of SMC1A mRNA expression Total RNA was extracted using TRIzol reagent according to the manufacturer’s instruction. 2ÃŽ ¼g total RNA was used to synthesize the first strand of cDNA using M-MLV Reverse Transcriptase. Real-time PCR reactions using Terraâ„ ¢ qPCR Direct SYBR ® Premix were run on Takara TP800-Thermal Cycler DiceTM Real-Time System. The following primers were used: SMC1A: 5’- AGCGAAAGGCAGAGATAATGG-3’ and 5’-GGTAGTCAAGAGGCAAGAAGG-3’; ÃŽ ²-actin: 5’- GTGGACATCCGCAAAGAC-3’ and 5’-AAAGGGTGTAACGCAACTA-3’. Thermal cycling condition were 1 min at 95 °C followed by 45 cycles of 95  °C for 5 s, 60 °C for 20 s, read absorbance value at the extension stage. The data was analyzed with Takara Thermal Dice Real Time System software Ver3.0. SMC1A relative mRNA levels was calculated using the 2-ΔΔCt method with normalization to ÃŽ ²-actin. And the conditions 2.7. Western blot analysis of SMC1A protein expression Cells were washed twice with ice-cold PBS and suspended in cell lysis buffer (2% Mercaptoethanol, 20% Glycerol, 4% SDS in 100mM Tris-HCl buffer, pH 6.8), and incubated for 15 min on ice. After centrifugation at 12,000 g for 15 min at 4oC, the supernatants were collected, and the protein content were measured using BCA protein assay kit. Equal amounts of protein were subjected to SDS-PAGE. After electrophoresis, blots were transferred onto PVDF membrane using an electro-blotting apparatus (Tanon, Shanghai, China). The membrane was blocked with TBST buffer containing 5% nonfat milk at room temperature for 1 h, and incubated with the primary antibodies in the blocking solution at 4oC overnight. After 3 washes with TBST buffer, the membrane was incubated with horseradish peroxidase (HRP) -conjugated secondary antibody (1:5000) at room temperature for 1 h. The signals of detected proteins were visualized by Pierce ECL western blotting detection kit (thermo scientific, USA). GAPDH protein level was used as an internal control to verify equal protein loading. 2.8. MTT assay Cell proliferation was evaluated by MTT assay. Exponential growth phase cells were plated at a final concentration of 2000 cells/well in 96-well plates and cultured for five consecutive days. MTT (10 µl, 10mg/ml) was then added, followed by incubation for another 4 hr at 37oC under humidified 5% CO2 atmosphere. The MTT was removed and addition of 150ÃŽ ¼l DMSO. Optical density (OD) of each well was measured at 490 nm using an ELx808 Absorbance Reader (Bio-Tek Instruments, USA). 2.9. Colony formation assay Cell growth and survival ability was also determined by the plate-colony-formation assay. In brief, 200 transfected cells were plated in 6-well plates. Cells were cultured for 14 days at 37oC under humidified 5% CO2 atmosphere. Culture medium was changed at 3-day intervals. Afterward, cells were incubate in 4% paraformaldehyde for 30 min at room temperature. The colonies were stained with Giemsa for 15 min, then washed with ddH2O and air-dried. The number of colonies (>50 cells/colony) was counted. 2.10. Flowcytometry Analysis Cell cycle distribution was assessed by propidium iodide (PI) staining. Briefly, the transfectedcells were harvested by trypsinization, centrifuged at 250 g for 5 min, washed twice with ice-cold PBS, and fixed in 70% ethanol at 4oC or -20oC for at least 1 h. Cells were collected and resuspended in PBS containing 100ÃŽ ¼g/ml RNase A and 40ÃŽ ¼g/ml PI, and then incubated at 4oC for 30 min, in dark. Cells were analyzed by flow cytometry using a FACSCalibur flow cytometer (Becton-Dickinson, San Jose, CA). The percentage of the cells in sub-G1, G0/G1, S, and G2/M phases were analyzed using ModFit (Verity Software House, Maine, USA) software. 2.11. Migration assay To explore the effect of SMC1A in the migration of prostate cancer cells, 24-well transwell chamber with 8.0ÃŽ ¼m pore polycarbonate filter inserts (Corning, cat no. #3422) was performed. In the upper chamber of each transwell, cells were suspended in serum-free F-12 containing 0.2% BSA. And F-12 supplemented with 10% FBS was added in each lower chamber. Then, the inserts were incubated at a 37 °C, 5% CO2/95% air incubator for overnight and the cells that had not penetrated the filters were removed. The migrated cells attached to the bottom side were fixed in 4% paraformaldehyde for 10 min and stained in0.1% crystal violet for 30 min, rinsed in PBS and examined under a bright-field microscope. 2.12. Tumorigenesis assay The influence of SMC1A silence on the tumor development of prostate tumor in vivo was examined. DU145, DU145-Lv-shCon or DU145 Lv-shSMC1A at 5 Ãâ€" 106 per mouse were injected subcutaneously into 4 weeks old Balb/c nude mice (n = 10 per group, Shanghai Laboratory Animal Center, Chinese Academy of Sciences, China). The development and growth of solid tumors were monitored by measuring tumor size using a vernier caliper in a blinded fashion every five days for a 27-days period. The tumor volume was calculated using a standard formula: tumor volume (mm3) = width (mm)2Ãâ€"length (mm)Ãâ€"0.5. At the end of the experiment, all mice were sacrificed and individual tumor weights were measured using a electronic balance. All the animal experiments were approved by the Animal Care Committee of the Second Military Medical University. 2.13. Statistical analysis The statistical analyses were performed with Graphpad Prism 5.0 software. The values are expressed as the mean of at least three different experiments  ± S.D. The results were analyzed by Student’s t-test, and P 3. Results 3.1. Expression of SMC1A in prostate cancer tissue and prostate cancer cells. To study the function of SMC1A in prostate cancer, we first analyzed its expression pattern in prostate cancer tissues. As shown in Fig 1A, SMC1A was strongly stained in prostate cancer tissue with a clear subcellular localization in the cytoplasm and nucleus of abnormal prostate epithelial cells. Then, the expression level of SMC1A was further analyzed by western blot, which showed that SMC1A was upregulated in prostate cancer tissues (Ca) compared to the adjacent normal tissues (N) (Fig 1B), implying a possible correlation between SMC1A and prostate cancer. To find a cell model for further investigation of SMC1A’s function in prostate cancer, we first analyzed the expression of SMC1A in four commonly used prostate cancer cell lines. Both WB and qPCR indicated that SMC1A expressions were elevated in PC-3, DU145 and 22RV1 cells in comparison to the androgen-sensitive LNCap cells (Fig 1C and D), which had less aggressiveness than the other three cell lines. We also found that S MC1A expression level was negatively correlated to AR expression level (Fig 1C and E), suggesting that SMC1A might be correlated with the malignancy of prostate cancer and involved in AR signaling. 3.2. Lentivirus-mediated knockdown of SMC1A in prostate cancer cells. As PC-3 and DU145 cells expressed much higher levels of SMC1A, they were used for further investigation. Both PC-3 and DU145 cells were untreated or transducated with Lv-shCon or Lv-shSMC1A. The transduction efficiencies were above 90% in both cells confirmed by fluorescent microscope (Fig 2A). WB analysis demonstrated that Lv-shSMC1A efficiently knocked down SMC1A expressions in PC-3 and Du145 cells (Fig B and C). Q-PCR results indicated that SMC1A was down-regulated more than 80% and 90% in PC-3 and DU145cells respectively. 3.3 Down-regulation of SMC1A inhibited cell proliferation in prostate cancer cells. After confirming the knocked down efficiency of SMC1A, PC-3 and DU145 cells were analyzed for cell growth rate with MTT assay. As shown in Fig 3A and E, cells transducted with Lv-shSMC1A displayed suppressed growth rate in comparison to the control or Lv-shCon transducted cells. The cells were then seeded onto 6-well plates for the analysis of colony formation ability. PC-3 and DU145 cells transducted with Lv-shSMC1A formed colonies with much smaller sizes compared to the control and Lv-shCon transducted cells (Fig 3B and F).The colonies formed in 6-well plates were photographed (Fig 3C and G), and counted (Fig 3D and H). The results suggested that both PC-3 and Du145 cells showed impaired colony formation abilities after SMC1A knockdown,indicating a pivotal role of SMC1A in regulation of prostate cancer cells proliferation.

IP addressing scenario Essay

When is it worth setting up a DHCP server for your office and how many computers would you say is the minimum amount that really justifies the work of setting up the server? Let’s first explore what DHCP is so we can understand what it does and can accurately answer the question at hand. DHCP stands for Dynamic Host Configuration Protocol and provides a mechanism for conveniently assigning network addresses and other network configuration parameters to a system upon request, usually when it is first bootstrapped or attached to the network. This is what allows you to manage the IP address of any given computer on your network from one location. Being that we ran out of addresses for IPv4 and are now basically only using borrowed addresses Maintaining the IP address for a network individually would be a huge chore for any network professional. Thankfully DHCP was invented making our job a little easier. Now the question is when is it worth setting up a DHCP server? In my opinion that would be, with anything larger than 2 computers. I am going to want to be able to manage the computers on my network from one location, my location. I am not going to want to have to get up from my desk, interrupting my current task to configure your IP address while making sure the IP address I am assigning isn’t a duplicate. This basically comes down to how much work you want to do as a network manager. Do you want to continually update and configure computers, at each individual desk, every time their IP address is released, or anytime you need to add computers to your office, or do you want to be able to do it from the comfort of your own computer? For me, I would be ok with 2 computers to maintain but any more then that and I am going to want the DHCP server set up.