Speaker identification and verification over short Essay Example For Students

Speaker identification and verification over short Essay distance telephone lines using artificial neural networksSPEAKER IDENTIFICATION AND VERIFICATION OVER SHORTDISTANCE TELEPHONE LINES USING ARTIFICIAL NEURALNETWORKSGanesh K Venayagamoorthy, Narend Sunderpersadh, and Theophilus N Andrewemailprotected emailprotected emailprotectedElectronic Engineering Department,M L Sultan Technikon,P O Box 1334, Durban, South Africa. ABSTRACTCrime and corruption have become rampant todayin our society and countless money is lost each yeardue to white collar crime, fraud, and embezzlement. This paper presents a technique of an ongoing workto combat white-collar crime in telephonetransactions by identifying and verifying speakersusing Artificial Neural Networks (ANNs). Resultsare presented to show the potential of this technique. We will write a custom essay on Speaker identification and verification over short specifically for you for only $16.38 $13.9/page Order now 1. INTRODUCTIONSeveral countries today are facing rampant crime andcorruption. Countless money is lost each year due towhite collar crime, fraud, and embezzlement. In todayscomplex economic times, businesses and individualsare both falling victims to these devastating crimes. Employees embezzle funds or steal goods from theiremployers, then disappear or hide behind legal issues. Individuals can easily become helpless victims ofidentity theft, stock schemes and other scams that robthem of their moneyWhite collar crime occurs in the gray area where thecriminal law ends and civil law begins. Victims ofwhite collar crimes are faced with navigating a dauntinglegal maze in order to effect some sort of resolution orrecovery. Law enforcement is often too focused oncombating street crime or does not have the expertiseto investigate and prosecute sophisticated fraudulentacts. Even if criminal prosecution is pursued, a criminalconviction does not mean that the victims of fraud areable to recover their losses. They have to rely on thcriminal courts awarding restitution after the convictionand by then the perpetrator has disposed of or hiddemost of the assets available for recovery. From the civillaw perspective, resolution and recovery can just be adifficult as pursuing criminal prosecution. Perpetratorsof white collar crime are often difficult to locate andserved with civil process. Once the perpetrators havebeen located and served, proof must be provided thatthe fraudulent act occurred and recovery/damages areneeded. This usually takes a lengthy legal fight, whichoften can cost the victim more money than the frauditself. If a judgement is awarded, then the task ofcollecting is made difficult by the span of time passedand the perpetrators efforts to hide the assets. Oftenafter a long legal battle, the victims are left with aworthless judgement and no recovery. One solution to avoid white collar crimes and shortenthe lengthy time in locating and serving perpetratorswith a judgement is by the use of biometrics techniquesfor identifying and verifying individuals. Biometrics aremethods for recognizing a user based on his/her uniquephysiological and/or behavioural characteristics. Thesecharacteristics include fingerprints, speech, face, retina,iris, hand-written signature, hand geometry, wrist veins,etc. Biometric systems are being commerciallydeveloped for a number of financial and securitapplications. Many people today have access to their companysinformation systems by logging in from home. Also,internet services and telephone banking are widely usedby the corporate and private sectors. Therefore toprotect ones resources or information with a simplepassword is not reliable and secure in the world oftoday. The conventional methods of using keys, accesspasswords and access cards are being easily overcomeby people with criminal intention. Voice signals as a unique behavioral characteristics isproposed in this paper for speaker identification andverification over short distance telephone lines usingartificial neural networks. This will address the whitecollar crimes over the telephone lines. Speakeridentification 1 and verification 2 over telephonelines have been reported but not using artificial neuralnetworks. Artificial neural networks are intelligent systems thatare related in some way to a simplified biological modelof the human brain. Attenuation and distortion of voicesignals exist over the telephone lines and artificialneural networks, despite a nonlinear, noisy andunstationary environment, are still good at recognizingand verifying unique characteristics of signals. Multilayerperceptron (MLP) feedforward neural networkstrained with backpropagation algorithm have beenapplied to identify bird species using recordings ofbirdsongs 3. Speaker identification based on directvoice signals using different types of neural networkshave been reported 4,5. The work reported in thispaper extends the work reported in 5 to short distancetelephone networks using ANN architectures describedin section 4 of this paper. The feature extraction, the neural network architecturesand the software and hardware involved in thedevelopment of the speaker identification andverification system are described in this paper. Resultswith success rates up to 90% in speaker identificationand verification over short distance telephone linesusing artificial neural networks is reported in this paper. 2. SPEAKER IDENTIFICATION ANDVERIFICATION SYSTEMA block diagram of a conventional speakeridentification/verification system is shown in figure 1. The system is trained to identify a persons voice byeach person speaking out a specific utterance into themicrophone. The speech signal is digitized and somedigital signal processing is carried out to create atemplate for the voice pattern and this is stored inmemory. The system identifies a speaker by comparing theutterance with the respective template stored in thmemory. When a match occurs the speaker is identified. The two important operations in an identifier are theparameter extraction and pattern matching. In parameteextraction distinct patterns are obtained from theutterances of each person and used to create a template. In pattern matching, the templates created in theparameter extraction process are compared with thosestored in memory. Usually correlation techniques areemployed for traditional pattern matching. ADC ParameterExtractionPatternMatchingMemoryTemplateOutputDevicemicFigure 1: Block Diagram of a Conventional SpeakerIdentification/Verification System. .u6ac99a9a77b678142032b6a77f6f2b44 , .u6ac99a9a77b678142032b6a77f6f2b44 .postImageUrl , .u6ac99a9a77b678142032b6a77f6f2b44 .centered-text-area { min-height: 80px; position: relative; } .u6ac99a9a77b678142032b6a77f6f2b44 , .u6ac99a9a77b678142032b6a77f6f2b44:hover , .u6ac99a9a77b678142032b6a77f6f2b44:visited , .u6ac99a9a77b678142032b6a77f6f2b44:active { border:0!important; } .u6ac99a9a77b678142032b6a77f6f2b44 .clearfix:after { content: ""; display: table; clear: both; } .u6ac99a9a77b678142032b6a77f6f2b44 { display: block; transition: background-color 250ms; webkit-transition: background-color 250ms; width: 100%; opacity: 1; transition: opacity 250ms; webkit-transition: opacity 250ms; background-color: #95A5A6; } .u6ac99a9a77b678142032b6a77f6f2b44:active , .u6ac99a9a77b678142032b6a77f6f2b44:hover { opacity: 1; transition: opacity 250ms; webkit-transition: opacity 250ms; background-color: #2C3E50; } .u6ac99a9a77b678142032b6a77f6f2b44 .centered-text-area { width: 100%; position: relative ; } .u6ac99a9a77b678142032b6a77f6f2b44 .ctaText { border-bottom: 0 solid #fff; color: #2980B9; font-size: 16px; font-weight: bold; margin: 0; padding: 0; text-decoration: underline; } .u6ac99a9a77b678142032b6a77f6f2b44 .postTitle { color: #FFFFFF; font-size: 16px; font-weight: 600; margin: 0; padding: 0; width: 100%; } .u6ac99a9a77b678142032b6a77f6f2b44 .ctaButton { background-color: #7F8C8D!important; color: #2980B9; border: none; border-radius: 3px; box-shadow: none; font-size: 14px; font-weight: bold; line-height: 26px; moz-border-radius: 3px; text-align: center; text-decoration: none; text-shadow: none; width: 80px; min-height: 80px; background: url(https://artscolumbia.org/wp-content/plugins/intelly-related-posts/assets/images/simple-arrow.png)no-repeat; position: absolute; right: 0; top: 0; } .u6ac99a9a77b678142032b6a77f6f2b44:hover .ctaButton { background-color: #34495E!important; } .u6ac99a9a77b678142032b6a77f6f2b44 .centered-text { display: table; height: 80px; padding-left : 18px; top: 0; } .u6ac99a9a77b678142032b6a77f6f2b44 .u6ac99a9a77b678142032b6a77f6f2b44-content { display: table-cell; margin: 0; padding: 0; padding-right: 108px; position: relative; vertical-align: middle; width: 100%; } .u6ac99a9a77b678142032b6a77f6f2b44:after { content: ""; display: block; clear: both; } READ: HIV: The Search For A Vaccine EssayThe speaker identification/verification system overtelephone lines investigated in this paper using artificialneural networks is shown in figure 2. FeatureExtractionNeural NetworkClassificationSpeaker IdentityorSpeaker AuthenticityTelephoneSpeech SignalFigure 2: Block Diagram of the SpeakerIdentification/Verification System using an ANN. In this paper, the speaker identification/verificationsystem reported is a text-dependent type. The system istrained on a group of people to be identified by eachperson speaking out the same phrase. The voice isrecorded on a standard 16-bit computer sound card fromthe telephone handset receiver. Although the frequencof the human voice ranges from 0 kHz to 20 kHz, mostof the signal content lies in the 0.3 kHz to 4 kHz range. The frequency over the telephone lines is limited to 0.3kHz to 3.4 kHz and this is the frequency band of interestin this work. Therefore, a sampling rate of 16 kHzsatisfying the Nyquist criterion is used. The voices arestored as sound files on the computer. Digital signalprocessing techniques are used to convert these soundfiles to a presentable form as input vectors to a neuralnetwork. The output of the neural network identifiesand verifies the speaker in the group. 3. FEATURE EXTRACTIONThe process of feature extraction consists of obtainingcharacteristic parameters of a signal to be used toclassify the signal. The extraction of salient features is akey step in solving any pattern recognition problem. Fospeaker recognition, the features extracted from aspeech signal should be consistent with regard to thedesired speaker while exhibiting large deviations fromthe features of an imposter. The selection of speakeruniquefeatures from a speech signal is an ongoingissue. Findings report that certain features yield betteperformance for some applications than do otherfeatures. Ref. 5 have shown on how the performancecan be improved by combining different types offeatures as inputs to an ANN classifier. Speaker identification and verification over telephonenetwork presents the following challenges:a) Variations in handset microphones which result insevere mismatches between speech data gatheredfrom these microphones. b) Signal distortions due to the telephone channel. c) Inadequate control over speaker/speakingconditions. Consequently, speaker identification and verificationsystems have not yet reached acceptable levels ofperformance over the telephone network. Severalfeature extraction techniques are explored but only thPower Spectral Densities (PSDs) based technique isreported in this paper. The discrete Fourier transform ofthe telephone voice samples is obtained and the PSDsare computed. The PSDs of three different speakers A,B and C uttering the same phrase is shown in figures 3,4 and 5 respectively. 0 1000 2000 3000 4000 5000 6000 7000 8000-80-60-40-20Power Spectrum Magnitude (dB)Frequency HzFigure 3: PSD of Speaker A0 1000 2000 3000 4000 5000 6000 7000 8000-100-80-60-40-20Power Spectrum Magnitude (dB)Frequency HzFigure 4: PSD of Speaker B0 1000 2000 3000 4000 5000 6000 7000 8000-150-100-50Power Spectrum Magnitude (dB)Frequency HzFigure 5: PSD of Speaker CIt can be seen from these figures that the PSDs of thspeakers differ from each other. Ref. 5 has reportedsuccess on speaker identification up to 66% and 90%with PSDs as input vectors to multilayer feedforwardneural networks and Self-Organizing Maps ( SOMs)respectively. 4. PATTERN MATCHING USING ARTIFICIALNEURAL NETWORKSArtificial Neural Networks (ANNs) are intelligentsystems that are related in some way to a simplifiedbiological model of the human brain. They arecomposed of many simple elements, called neurons,operating in parallel and connected to each other bysome multipliers called the connection weights orstrengths. Neural networks are trained by adjustingvalues of these connection weights between theneurons. Neural networks have a self learning capability, arefault tolerant and noise immune, and have applicationsin system identification, pattern recognition,classification, speech recognition, image processing,etc. In this paper, ANNs are used for pattern matching. The performance of different neural networarchitectures are investigated for this application. Thipaper presents results for the MLP feedforward networkand the self-organizing feature map. Descriptions ofthese networks are given below. 4.1. MLP FEEDFORWARD NETWORKA three layer feedforward neural network with asigmoidal hidden layer followed by a linear output layeis used in this application for pattern matching. Theneural network is trained using the conventionalbackpropagation algorithm. In this application, anadaptive learning rate is used; that is, the learning rate isadjusted during the training to enhance faster globalconvergence. Also, a momentum term is used in thebackpropagation algorithm to achieve a faster globalconvergence. The MLP network in figure 6 is constructed in theMATLAB environment 6. The input to the MLPnetwork is a vector containing the PSDs. The hiddenlayer consists of thirty neurons for four speakers. Thenumber of neurons in the output layer depends on thenumber of speakers and in this paper it is four. sigmoidal activation functionlinear activation function1st speakerNth speakerVectorof PSDs Figure 6: MLP NetworkAn initial learning rate, an allowable error and themaximum number of training cycles/epochs are theparameters that are specified during the training phaseto the MATLAB neural network program. 4.2. SELF-ORGANIZING FEATURE MAPSThe second type of neural network selected for thisinvestigation is the self-organizing feature map 7. Thisneural network is selected because of its ability to learna topological mapping of an input data space into apattern space that defines discrimination or decisionsurfaces. The operation of this network resembles theclassical vector-quantization method called the k-meansclustering. Self-organizing feature maps are moregeneral because topologically close nodes are sensitiveto inputs that are physically similar. Output nodes willbe ordered in a natural manner. .ue4fbbef56c46457914065d1eaf78aa15 , .ue4fbbef56c46457914065d1eaf78aa15 .postImageUrl , .ue4fbbef56c46457914065d1eaf78aa15 .centered-text-area { min-height: 80px; position: relative; } .ue4fbbef56c46457914065d1eaf78aa15 , .ue4fbbef56c46457914065d1eaf78aa15:hover , .ue4fbbef56c46457914065d1eaf78aa15:visited , .ue4fbbef56c46457914065d1eaf78aa15:active { border:0!important; } .ue4fbbef56c46457914065d1eaf78aa15 .clearfix:after { content: ""; display: table; clear: both; } .ue4fbbef56c46457914065d1eaf78aa15 { display: block; transition: background-color 250ms; webkit-transition: background-color 250ms; width: 100%; opacity: 1; transition: opacity 250ms; webkit-transition: opacity 250ms; background-color: #95A5A6; } .ue4fbbef56c46457914065d1eaf78aa15:active , .ue4fbbef56c46457914065d1eaf78aa15:hover { opacity: 1; transition: opacity 250ms; webkit-transition: opacity 250ms; background-color: #2C3E50; } .ue4fbbef56c46457914065d1eaf78aa15 .centered-text-area { width: 100%; position: relative ; } .ue4fbbef56c46457914065d1eaf78aa15 .ctaText { border-bottom: 0 solid #fff; color: #2980B9; font-size: 16px; font-weight: bold; margin: 0; padding: 0; text-decoration: underline; } .ue4fbbef56c46457914065d1eaf78aa15 .postTitle { color: #FFFFFF; font-size: 16px; font-weight: 600; margin: 0; padding: 0; width: 100%; } .ue4fbbef56c46457914065d1eaf78aa15 .ctaButton { background-color: #7F8C8D!important; color: #2980B9; border: none; border-radius: 3px; box-shadow: none; font-size: 14px; font-weight: bold; line-height: 26px; moz-border-radius: 3px; text-align: center; text-decoration: none; text-shadow: none; width: 80px; min-height: 80px; background: url(https://artscolumbia.org/wp-content/plugins/intelly-related-posts/assets/images/simple-arrow.png)no-repeat; position: absolute; right: 0; top: 0; } .ue4fbbef56c46457914065d1eaf78aa15:hover .ctaButton { background-color: #34495E!important; } .ue4fbbef56c46457914065d1eaf78aa15 .centered-text { display: table; height: 80px; padding-left : 18px; top: 0; } .ue4fbbef56c46457914065d1eaf78aa15 .ue4fbbef56c46457914065d1eaf78aa15-content { display: table-cell; margin: 0; padding: 0; padding-right: 108px; position: relative; vertical-align: middle; width: 100%; } .ue4fbbef56c46457914065d1eaf78aa15:after { content: ""; display: block; clear: both; } READ: Research Paper on Chewing Tobacco EssayTypically, the Kohonen feature map consists of a twodimensional array of linear neurons. During the trainingphase the same pattern is presented to the inputs of eachneuron, the neuron with the greatest output value isselected as the winner, and its weights are updatedaccording to the following rule:w t w t x t w t i i i ( ) () () () + = + ;#8722; 1 a (1)where wi(t) is the weight vector of neuron i at time t, is the learning rate and x(t) is the training vector. Those neurons within a given distance, theneighborhood, of the winning neuron also have theirweights adjusted according to the same rule. Thisprocedure is repeated for each pattern in the training setto complete a training cycle or an epoch. The size of theneighborhood is reduced as the training progresses. Inthis way the network generates over many cycles anordered map of the input space, neurons tending tocluster together where input vectors are clustered,similar input patterns tending to excite neurons insimilar areas of the network. 5. IMPLEMENTATION OF THE SPEAKEIDENTIFICATION AND VERIFICATION SYSTEMThe work that is being reported in this paper isimplemented in software. The telephone speech icaptured and processed on a Pentium II 233 MHzcomputer with a 16 bit sound card. The telephonereceiver is interfaced to the sound card. Telephonspeech is captured over signals transmitted within 10kilometres of transmission network. Digital signalprocessing and neural network implementations arecarried out using the MATLAB signal processing andneural network toolboxes respectively. This work iscurrently undergoing and an implementation of a realtimespeaker identification and verification system ovetelephone lines on a digital signal processor isenvisaged. 6. EXPERIMENTAL RESULTSThe MLP network is trained with the PSDs of eightvoice samples recorded at different instants of timeunder controlled and uncontrolled speaking conditionsof four different speakers uttering the same phrase at alltimes. Controlled speaking conditions refer to noise anddistortion free conditions unlike uncontrolled speakingconditions which have noise and distortion on thetransmission lines. The number of PSD points for eachvoice sample is about 500. As mentioned in section 4.1,an adaptive learning rate is used for the MLP network. The initial learning rate is 0.01. The allowable sumsquared error and maximum number of epochsspecified to the MATLAB neural network program i0.01 and 10000 respectively. It is found that the sumsquared error goal is reached within 1000 epochs. A success rate of 100% is achieved when the trainedMLP network is tested with the same samples used inthe training phase. However, when untrained samplesare used, only a 63% success rate is obtained. This isdue to the inconsistency in the PSDs of the inputsamples with those used in the training phase. The MLPnetwork is also tested with unseen voice samples ofpeople who are not included in the training set and thenetwork successfully classified these voice samples asunidentified. Four speakers are identified using the self-organizingfeature map like in the case of the MLP network. Aninitial learning rate of 0.01, an allowable sum squarederror of 0.01 and a maximum of 70000 epochs arespecified at the start of the training process to theMATLAB neural network program. The results with theself-organizing feature map shows a drastic change inthe success rate in identifying the speakers as reportedin 5. With PSDs as inputs, a success rate of 85% and90% is achieved under uncontrolled and controlledspeaking conditions respectively. Ref.5 has reported that success rate can be increasedto 98% under uncontrolled speaking conditions byusing Linear Prediction Coefficients (LPCs) as inputs toSOMs which remains to be yet to be tried out in thiswork. Currently, with the PSDs as inputs a lot ofcomputations is involved and the SOM takes a lot oftime to learn. 7. CONCLUSIONSThis paper has reported on the feasibility of usingneural networks for speaker identification andverification over short distance telephone lines and hashown that performance with the self-organizing map ishigher compared to that with the multilayer feedforwardneural network. Different feature inputs to the selforganizingmap remains to be tried out in order toachieve higher identification/verification ratesminimizing the training time and the size of thenetwork. Speaker identification with telephone speechsignals over long distance telephone lines is currentlbeing investigated using similar techniques. This paper has shown that speaker identification ispossible over the telephone lines and thereforetelephonic bank and other transactions can beauthenticated. Hence a technique to combat and/orreduce white collar crime. 8. REFERENCES:1 D.A.Reynolds, Large population speakeidentification using clean and telephone speech, IEEESignal Processing Letters, vol. 2 no. 3 March 1995, pp. 46 48. 2 J.M.Naik, L.P.Netsch, G.R.Doddington, Speakerverification over long distance telephone lines,Proceedings of IEEE International Conference onAcoustics, Speech, and Signal Processing (ICASSP),23-26 May 1989, pp. 524 527. 3 A.L.Mcilraith, H.C.Card, Birdsong RecognitionUsing Backpropagation and Multivariate Statistics,Proceedings of IEEE Trans on Signal Processing, vol. 45, no. 11, November 1997. 4 G.K.Venayagamoorthy, V.Moonasar,K.Sandrasegaran, Voice Recognition Using NeuralNetworks, Proceedings of IEEE South AfricanSymposium on Communications and Signal Processing(COMSIG 98), 7-8 September 1998, pp. 29 32. 5 V.Moonasar, G.K.Venayagamoorthy, Speakeridentification using a combination of differentparameters as feature inputs to an artificial neuralnetwork classifier, accepted for publication in theProceedings of IEEE Africon 99 conference, CapeTown, 29 September 2 October 99. 6 H.Demuth, M.Beale, MATLAB Neural NetworkToolbox Users Guide, The Maths Works Inc., 1996. 7 T.Kohonen, Self-organizing and associate memorySpring Verlag, Berlin, third edition, 1989.

Becoming a Successful Computer Programmer essays

Becoming a Successful Computer Programmer essays The career I choose to research is computer programming. Computer programmers write and maintain detailed instructions used to create software. These instructions, also called code, are made up of different languages that a computer can recognize. Because there are many language a programmer always needs to be able to learn new things. Computers have always been a hobby for me and satisfy many needs that I require in a career. In order to be a successful programmer, there are many skills you must possess. The most important skill is logic. Because of all the detail and explicit instructions required to program software a person must be able to think logically. In high school my strongest class was math. Math is my favorite subject because it deals with breaking things down and simplifying them. This is what programming is all about. Another quality I have that relates to programming is patients. About 80 percent of programmers work time spent debugging. Debugging is the process of finding all the errors in the code and fix them. This requires tons of patients because many of these errors are hard to seek out. Also, I love to solve problems. In high school, my computer science teacher would give us brain teasers, and I would sit up all night until I figured it out. A computer programmer must be able to receive a problem and find the most efficient way to solve it. The three qualities I looked at in choosing a career are salary, interesting work, and being able to see that my work has created something. People say that money isn't everything, but it certainly does help. In 1998 the median of annual earnings for a computer programmer was $47,550. Half of these people earned between $37,000 and $70,500 a year. These numbers are rising due to a growing demand for programmers. Working with computers is almost always fun and exciting for me. There is something about working with one of the greatest inventions of ...

Export Feasibility Analysis of a Consumer Item to a Foreign Country Essay

Export Feasibility Analysis of a Consumer Item to a Foreign Country - Essay Example There are no barriers to exports from the United States of America to Poland. There is however a drop in exports from the United States of America to Poland. The domestic production of telecommunication products is deficient and Poland depends on imports. In 1999 $124 million worth of telecommunication equipment was imported from the United States of America, but in 2000 these exports dropped by thirty five percent because of increasing competition from Western European countries. (Telecommunications. Poland). Competition in the field of cellular communications is fierce and it is this competitive activity that has seen the growth in the use of mobile phones in Poland. (Poland: Telecoms and Technology Background). Indirect competition comes from fixed line telephones and the status of the fixed line infrastructure in Poland is the driving force behind the growth of use of mobile phones and the penetration that is seen in Poland. Though Poland is in the front of the countries in the c entral European region from the point of view of service revenue, the fixed line penetration is just the opposite. The inference that can be easily made from this is that demand far outstrips supply and this situation has developed from the monopoly of TPSA till 2003. Though the situation has changed since then, indirect competition does not seem a stumbling block for the growth in the use of mobile phones in Poland. This is the area of fierce competition that has seen an increase in the potential and penetration in cellular communications in Poland.

D1213 Assignment Example | Topics and Well Written Essays - 500 words

D1213 - Assignment Example Production of bottled water is under strict regulations and monitoring prescribed by FDA which ensures cleanliness and purity of the water. In case of tap water most of the time water lines are affected due to contamination which can be avoided by using bottled water. As there are, both negative and positive aspects related with bottled water so according to me it is better to use bottled water from a reputed manufacturer (Azoulay, Garzon and Eisenberg). Before consume bottled water it is always recommended that to verify the content level and make sure that it is purified according to FDA regulations. Ergogenic aids are mainly the supplements, which has a quality to enhance performance for a certain time by increasing energy utilization. The external influence provided by ergogenic aids is associated with not only performance but also can remove certain restraints, which can limit capacity of an individual (Tokish, Kocher and Hawkins). International Olympic committee has an independent body of that is World anti drug doping agency (WADA) has a prescribed guidelines for ergogenic aids and there mode of legality. Blood doping is a kind of ergogenic aids, which is illegal as it cause severe side effects in human body. There are also some supplements also which are permissible according to the WADA. Supplements such as creatine, caffeine, amino acids are legal in a permissible range. There following questions can be asked to an athlete before taking this kind of supplement, Further research needs to be done on the effect on performance and side effects of the supplements. Athletes and their coaches who are using these kinds of supplements need proper counseling. Awareness program can be organized to give a proper picture of the side effects. Most importantly Legal action should be taken to the users of banned supplements. Tokish, John M, Mininder S Kocher, and Richard J Hawkins. â€Å"Ergogenic Aids: A Review Of Basic Science,

Analysis Essay Example | Topics and Well Written Essays - 500 words - 24

Analysis - Essay Example He may utilize a wide range of gadgets to help him do this—viewpoint is one of them—yet in any case reality around a complete visual impression relies on upon one thing, truth of tone. Drawing may be rundown, hues dull, yet in the event that the relations of tone are genuine, the picture will hold. Profundity and measurement are rendered by the utilization of straight point of view, by the covering of the layers of shapes, and specifically, through the utilization of tone. This compositional component works inside the picture in various ways. First and foremost, there is the presence of characteristic light inside the painted room and past it. The pictorial space in the midground and forefront is lit from two sources: by meager shafts of light from the open entryway, and by expansive streams getting through the window to one side. The light from the window enlightens both the studio closer view and the unrepresented territory before it, in which the ruler, the ruler, and the viewer are attempted to be situated. Light partitions the scene into three unmistakable parts, with frontal area and foundation planes firmly lit up, between which an obscured halfway space incorporates silhouetted figures. Velà ¡zquez utilizes this light not just to add volume and definition to every structure additionally to characterize the central purposes of the sketch. As the light streams in from the right it splendidly glimmers on the plait and brilliant hair of the female diminutive person, who is closest the light source. But since her face is turned from the light, and in shadow, its tonality does not make it a state exceptionally compelling. Likewise, the light looks diagonally on the cheek of the woman in-holding up close to her, however not on her facial highlights. A lot of her delicately hued dress is diminished by shadow. The Infanta, be that as it may, remains in full enlightenment, and with her face turned towards the light source, despite the fact that

Rapamycin And Cisplatin In Breast Cancers

Rapamycin And Cisplatin In Breast Cancers Recent gene expression profiling has identified five breast cancer subtypes, of which the basal-like breast cancers are the most aggressive and possess serious clinical challenges as there are currently no targeted therapies available. Although there is increasing evidence that these tumors confer specific sensitivity to cisplatin, its success is often compromised due to its dose-limiting nephrotoxicity and development of drug resistance. To overcome this limitation, our focus is to maximize the benefits associated with cisplatin therapy through drug combination strategies. Using a well-validated kinase inhibitors library, we showed that inhibition of mTOR, TGFà ¯Ã‚ Ã‚ ¢RI, NFà ¯Ã‚ Ã‚ «B, PI3K/AKT and MAPK pathway sensitized the basal-like MDA-MB-468 cells to cisplatin treatment. Further evaluation demonstrated that combination of mTOR inhibitor, rapamycin, and cisplatin generated significant drug synergism specifically in basal-like cells (MDA-MB-468, MDA-MB-231 and HCC1937). Th ese synergistic effects were not observed in the luminal-like T47D and MCF-7 cells. We further showed that the synergistic effects of rapamycin and cisplatin is mediated through p73. Treatment of rapamycin induced p73 upregulation and synergized cisplatin activity through activation of the p73 pathway. Depletion of endogenous p73 in basal-like cells abolished these synergistic effects suggesting that p73 is required for the rapamycin and cisplatin synergism. In conclusion, combination of mTOR inhibitors and cisplatin may be a useful therapeutic strategy in basal-like breast cancers. INTRODUCTION Recent identification of novel breast cancer subtypes with distinct biological features promises a more specific, effective and less toxic therapies to the patients. Through gene profiling, breast cancer can be categorized into five different subtypes with distinct clinical outcome. The five major subtypes of breast cancer are luminal A, luminal B, human epidermal growth factor receptor-2 (HER2) overexpressing, normal-like and basal-like breast cancer [1, 2]. Of particular important is the basal-like breast cancer which accounts for 15-20% of breast cancers overall and confers a remarkably poor prognosis compared to other subtypes. Majority of basal-like breast cancers exhibit a triple-negative phenotype, characterized by the lack of expression of estrogen receptor (ER), progesterone receptor (PR) or HER2 amplification, and often have high frequency of p53 mutation [3, 4]. Due to the lack of expression of these receptors (ER, PR and HER2), patients with basal-like breast cancers usua lly do not response to hormonal therapy, Herceptin or chemotherapy [5, 6]. As a consequence, the mortality rate of basal-like breast cancer is relatively high in comparison with the non-basal subtype [1]. Numerous clinical studies are currently ongoing to identify novel therapy for treatment of basal-like breast cancers. These include the use of specific targeted therapeutic agents (e.g. Cetuximab, Dasatinib, Bevacizumab, Abraxane and Erlotinib) or conventional chemotherapeutics agents (e.g. cisplatin, doxorubicin, and paclitaxel), either as single agent or in combination, as first line therapy for basal-like breast cancers [7-9]. Cisplatin, a chemotherapeutic agent not commonly used for breast cancer, come to light in the management of basal-like breast cancer on account of evidence that breast cancer cells with basal-like phenotype confer a selective sensitivity towards cisplatin as compared to other chemotherapeutic agents. A variety of evidence suggests that basal-like breast cancers may share defects in BRCA1-associated pathways, of which DNA repair mechanism has been compromised [10]. Indeed, recent clinical studies have demonstrated the clear advantage of cisplatin in treatment of basal-like breast cancer compared to other chemotherapeutic agents [11, 12]. Nevertheless, dose-limiting toxicity including nephrotoxicity, neurotoxicity and ototoxicity have withold the wide-spread use of cisplatin in treating breast cancers in the clinic. To address this problem, we developed a high-throughput screening assay to rapidly identify new therapeutic agents that could synergize the antitumor effects of cisplatin in basal-like breast cancers. Through the use of a small chemical library that targets some of the most relevant oncogenic pathways in basal-like breast cancer, we show that inhibition of mTOR by rapamycin incurred a specific synergistic effect with cisplatin in basal-like breast cancer cells. This synergistic effect is mediated in part through the induction and activation of p73 in the presence of rapamycin and cisplatin, respectively. Together, our findings demonstrate evidence of a synergistic relation between rapamycin and cisplatin in both inhibition of cell growth and induction of apoptosis. This suggests that rapamycin and cisplatin may be a rational combination of a targeted therapy for the refractory basal-like breast cancers. Materials and Methods Cell lines and cell culture The human breast carcinoma cell lines MCF-7, T47D, MDA-MB-231, MDA-MB-468 and HCC1937 were obtained from the American Type Culture Collection (Manassas, VA) and maintained in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 IU/ml penicillin and 100 ÃŽÂ ¼g/ml streptomycin (Sigma-Aldrich, St. Louis, MO, USA) at 37 °C with 5% CO2. MTT cell proliferation assay Dose-response curves and IC50 values were determined using the methyl thiazolyl tetrazolium (MTT) cell viability assay as described previously [4, 13]. Cells were seeded into 96-well plates for 24 hours at a density of 5 ÃÆ'- 103 cells/well. Serial drug dilutions were prepared in medium immediately before each assay, and viable cell masses following 3 days of drug exposure were determined by cell-mediated MTT reduction. Cell growth as well as drug activity was determined by measuring absorbance at 550 nm using an Anthos systems plate reader. Construction of IC50 mean graph The IC50 mean graph was constructed as defined by the Developmental Therapeutics Program of the National Cancer Institute (http://dtp.nci.nih.gov). The mean graph consists of positive (more sensitive) and negative (less sensitive) delta values, generated from a set of IC50 values by using a three-step calculation. The IC50 values for each of cell line against the tested compound were converted to log(IC50) values. For each tested compound, the log(IC50) values are averaged. Finally, the individual IC50 value is then subtracted from the average to generate the delta value. Positive delta values project to the right of the vertical line and represent cellular sensitivities to the test agent that exceed the mean. Negative values project to the left and represent cell line sensitivities to the test agent that are less than the average value. Library screening The Inhibitor Select„ ¢ chemical library which consists of 160 well-characterized, cell-permeable inhibitors was purchased from EMD Chemicals, USA. MDA-MB-468 cells at the logarithmic phase of growth were seeded into 96-well plate at a density of 5 ÃÆ'- 103 cells/well. Each compound was added to a final concentration of 10  µM in the absence or presence of 1  µM cisplatin. Plates were incubated for 72h at 37 °C. Cell proliferation was examined using MTT assay as described previously. Combination treatments that induce growth inhibition higher than those of the same doses used alone (p Drug interaction analysis Drug combination analysis was performed by using the method as described by Chou and Talalay [14]. Briefly, cells were seeded at 5 ÃÆ'- 103 cells/well in 96-well plates and treated with various concentrations of cisplatin and compound alone or in combination for 72h. Cell proliferation was measured in each well by MTT assay. Multiple drug dose-effect calculations and the combination index plots were generated using Calcusyn software (Biosoft, Cambridge, UK). Combination index, CI 1 indicate synergism, additive effect and antagonism, respectively. Apoptosis assays Quantitation of apoptosis by annexin V/PI staining was performed as described previously [3, 4]. Briefly, both floating and attached cells were collected 72h after drug treatments. Apoptotic cell death was determined using the BD ApoAlert annexin V-FITC Apoptosis Kit (BD Biosciences, USA) according to the manufacturers instructions, and cells were analyzed on a FACSCalibur flow cytometer using CellQuest Pro software (version 5.1.1; BD Biosciences, USA). Quantitative PCR (qPCR) analysis Total RNA from cells was extracted using Qiagen RNA isolation kit (Qiagen, Valencia, CA, USA) according to the manufacturers protocol. First-strand cDNA was synthesized from total RNA using random hexamer primers and the SuperScript II system for RT-PCR (Invitrogen, Carlsbad, USA). Gene expression levels were measured by qPCR using the iQ SYBR Green Supermix reagent and an Biorad iQ5 real-time PCR detector system (Bio-Rad, Richmond, CA, USA). Data analysis was performed using Opticon Monitor Analysis Software V1.08. The expression of each gene was normalized to ÃŽÂ ²2M as a reference. The relative copy numbers were calculated from an 8-point standard curve generated from a 10-fold serial dilution of full-length cDNA constructs as described previously [3, 4]. Specific forward and reverse primer sequences are as follows : TAp73fwd, 5-GCACCACGTTTGAGCACCTCT-3; TAp73rev, 5- GCAGATTGAACTGGGCCATGA-3; ÃŽÂ ²2Mfwd, 5-AGCTGTGCTCGCGCTACTCTC-3; ÃŽÂ ²2Mrev, 5-CACACGGCAGGCATACTCATC-3; PUMA fwd PUMArev NOXAfwd NOXArev. The conditions for all QRT-PCR reactions were as follows: 3 minutes at 94 °C followed by 40 seconds at 94 °C, 40 seconds at 60 °C, and 25 seconds at 72 °C for 40 cycles. All PCR products were confirmed by the presence of a single peak upon melting curve analysis and by gel electrophoresis. No-template (water) reaction mixtures and no-RT mixtures were performed on all samples as negative controls. All experiments were performed in duplicate. Protein isolation and Western blot analysis Protein lysates from cells were extracted in ice-cold lysis buffer (0.75% NP-40, 1 mM DTT, and protease inhibitors in PBS). Total protein (25 ÃŽÂ ¼g) was subjected to SDS-PAGE followed by immunoblotting with the following antibodies: p73 (diluted 1:1,000, Ab-2; CalBiochem); pS6K (diluted 1:1,000; Cell Signaling Technology); S6K (diluted 1:1,000, Ab9645; Abcam); and ÃŽÂ ²-tubulin (diluted 1:2,500, D-10; Santa Cruz Biotechnology). Lentiviral production and infection The shRNA lentiviral constructs were created by transferring the U6 promoter-shRNA cassette into a lentiviral backbone, and high-titre lentiviral stocks were generated by co-transfection with packaging vectors into 293T cells as described previously [3, 4, 13]. The shRNA target sequences for TAp73 was 5-GGATTCCAGCATGGACGTCTT-3. The TAp73 targeted sequence is found within p73 exon 3. Therefore, this shRNA does not target ΆNp73 [4]. RESULTS Selective sensitivity of basal-like breast cancer toward cisplatin To gain an overview of the selectivity of chemotherapeutic agents for basal-like breast cancer cells, we compared their antiproliferative properties in a panel of basal-like and luminal-like breast cancer cell lines which has been validated previously through gene profiling [15]. All cells were treated with increasing concentrations of cisplatin, paclitaxel or doxorubicin for 72 hours and growth measured using the MTT assay. Figure 1A and B summarizes the results from these breast cancer cell lines in which basal-like breast cancer cells demonstrated selective sensitivity to cisplatin. This selectivity was absence in cells treated with paclitaxel or doxorubicin suggesting that basal-like breast cancer cells confer selective sensitivity towards cisplatin (Figure 1A, B and Supplement Table 1). Small chemical library screening identify rapamycin as synergistic agents for cisplatin Although cisplatin is currently one of the most used agents in the treatment of cancer, the use of cisplatin is hampered by its side effects, especially neurotoxicity, nephrotoxicity and rug resistance [16]. Hence, the present study was aimed to identify chemosensitizers that could synergize the effects of cisplatin for treatment of basal-like breast cancers. To identify small molecules that enhance sensitivity of basal-like breast cancer cells to cisplatin, a cell-based high-throughput screen was performed using MDA-MB-468 cell line and a small chemical library consisting of 160 well validated specific inhibitors. The screens were done in 96-well plates to which compounds were added at 10  µM, followed by cisplatin at 1  µM. Cell viability was measured 72 hours later by MTT assay. Each plate included controls of untreated cells, cells treated with compounds or cisplatin only, and cells treated with a combination of both agents. Combinations of the treatments that induced growth inhibition higher than those of the same doses used alone (p The molecules identified in this screen includes rapamycin, [3-(Pyridin-2-yl)-4-(4-quinonyl)]-1H-pyrazole (LY364947), 4-(3-Chloroanilino)-6,7-dimethoxyquinazoline (AG1478), (E)3-[(4-Methylphenyl)sulfonyl]-2-propenenitrile (BAY11-7082), 2-(4-Morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002) and 4-(4-Fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole (SB203580). Structures of these compounds and their growth inhibitory effects were shown in Figure 2. The molecular target of these compounds was listed in Table 1. The 6 compounds identified are specific inhibitors of the mTOR, TGF-à ¯Ã‚ Ã‚ ¢, EGFR, NFà ¯Ã‚ Ã‚ «B PI3K/AKT and MAPK pathways which have been previously reported to be upregulated specifically in basal-like breast cancers [7, 17-20]. However, when tested for synergism with cisplatin at fixed concentration ratio, only rapamycin showed synergism when combined with cisplatin in MDA-MB-468 cells using the isobologram method that simulate the median-dose combination index (CI) [14]. The mean combination index at ED50, ED75, and ED90 of rapamycin (10:1 cisplatin:rapamycin) when combined with cisplatin was 0.52 à ¯Ã¢â‚¬Å¡Ã‚ ± 0.06, where combination index Specific synergistic effects of rapamycin and cisplatin in basal-like breast cancer cells Next, we sought to investigate if combination of cisplatin with rapamycin exhibit specific synergism in basal-like breast cancers by comparing their effects in a panel of breast cancer cell lines. The antiproliferative effect of treatment was evaluated using MTT assays and drug interaction was assessed by the isobologram method as described in the previous section. The results demonstrated that combination of rapamysin and cisplatin exhibited selective synergistic effects only in the basal-like MDA-MB-468, MDA-MB-231 and HCC1937 cells, but not in the luminal-like T47D or MCF-7 cells (Figure 3A and Table 2). To ensure that the lack of synergistic effects of rapamycin and cisplatin observed in luminal-like cells is not due the general lack of sensitivity of luminal-like cells to cisplatin per se, we compared the apoptotic effects of cisplatin and rapamycin alone or in combination in MDA-MB-231 and T47D cells using an equal potent dose of cisplatin (IC50 dose). Both cell lines exhibited similar amount of apoptosis following treatment with their respective IC50 dose of cisplatin. Interestingly, when both cell lines were treated with combination of cisplatin and rapamycin, synergistic effects were evidenced only in MDA-MB-231 cells but not in T47D cells. The potentiation effects of rapamycin observed in MDA-MB-231 cells were further enhanced by sequential treatment with rapamycin for 6 hours followed by cisplatin (data not shown). These results demonstrated that rapamycin is capable of sensitizing basal-like breast cancer cells to cisplatin, suggesting that the synergistic effects of rapamyc in and cisplatin may be mediated through a common pathway. Rapamycin sensitizes basal-like cells to cisplatin through p73 up regulation Previous study has shown that inhibition of mTOR by rapamycin up regulate p73 in breast cancer cells [21]. We and others have also demonstrated that p73 is overexpressed in a subset of triple-negative breast tumors and that p73 is required for cisplatin sensitivity in basal-like breast cancer cells [4]. On the basis of these findings, we ask if the synergistic effects of rapamycin and cisplatin combination could be explained by the activation of the p73 pathway. To test this notion, MDA-MB-231 cells were treated with 10  µM of cisplatin and 100 nM rapamycin alone or in combination for 48 hours. mTOR inhibition was confirmed using phospho-S6K as a marker of mTOR activity. Quantitation of the mRNA and protein expression of the TAp73 was performed using qPCR and immunoblotting, respectively. The results showed that treatment of rapamycin or cisplatin alone did not change the expression of TAp73 mRNA but induced significant up regulation of TAp73 protein expression in MDA-MB-231 cells (Figure 4A and B). Next, we evaluated the expression of the two potent pro-apoptotic BH3 only proteins, PUMA and NOXA, which has been identified as specific p73 target genes, using qPCR [22, 23]. As expected, treatment of cells with cisplatin for 48 hours induced mRNA expression of PUMA and NOXA in both MDA-MB-231 and MDA-MB-468 cells. Significant induction of PUMA and NOXA were also observed in MDA-MB-231 cells but not in MDA-MB-468 cells following treatment with rapamycin alone. When cells were treated with combination of cisplatin and rapamycin, the expression of PUMA and NOXA was further enhanced, corroborated with the massive induction of apoptosis as shown in Figure 3B. Together, these results suggest that rapamycin synergize cisplatin activity in basal-like cells through induction of p73 pathway. p73 is required for the synergistic effects of rapamycin and cisplatin in basal-like breast cancer cells To further evaluate whether p73 is the effector of the synergistic effects of rapamycin and cisplatin in basal-like breast cancer cells, we generated a series of isogenic cell lines that have been depleted for TAp73 by stably expressing a shRNA species that target specifically human TAp73. Unlike MDA-MB-231 cells which express one predominant TAp73 isoform (TAp73à ¯Ã‚ Ã‚ ¢) isoform, MDA-MB-468 cells express high levels of two p73 isoforms, TAp73à ¯Ã‚ Ã‚ ¡ and TAp73à ¯Ã‚ Ã‚ ¢ [21]. Figure 5A showed efficient knock-down of TAp73 isoforms in MDA-MB-231 and MDA-MB-468 cells. As expected, treatment of cisplatin alone induced significant amount of apoptosis in MDA-MB-468 vector control cells. This apoptotic effects were further enhanced in the presence of rapamycin, consistent with our previous observations (Figure 3B). In stark contrast, depletion of TAp73 not only reduced the amount of apoptosis following treatment of cisplatin alone but also completely abrogated the synergistic effects of rapamycin (Figure). This result is further supported by the isobologram analysis which showed a lack of synergism of rapamycin-cisplatin treatment in the TAp73 depleted cells. Together, these results suggest that TAp73 is required for the synergistic effects of rapamycin and cisplatin in basal-like breast cancers. DISCUSSION By gene profiling, breast cancers can be classified into 5 molecularly distinct subtypes: luminal A, luminal B, HER2+, basal-like and normal breast cancers. The basal-like subtype, which represents 15-20% of breast cancers, has been subjected to extensive investigation in recent years due to its association with poor patient survival [1, 2, 20, 24]. Unlike many breast cancers, patients diagnosed with basal-like breast cancers are not eligible for molecular targeted therapy that target ER (e.g. tamoxifen, aromatase inhibitors) or HER2 (e.g. Herceptin) as they do not express the estrogen receptor (ER) or progesterone receptor (PR), nor do they have amplified HER2 [1, 24]. The treatment option therefore is relied on aggressive conventional chemotherapies which have limited efficacy, many side effects and often high rate of relapse. Hence, development of an effective therapeutic strategy remains an important goal in the management of basal-like breast cancer. Several lines of evidence has suggested a link between basal-like breast cancers and BRCA1 deficiency [7, 25, 26]. In most cases, the clinical features and outcomes for women with sporadic basal-like breast cancers are broadly similar to those with BRCA1-related cancers including high tendency of developing high grade, high mitotic index tumors, shorter time of relapse, similar pattern of metastatic spread and cytogenetic changes associated with frequent loss of X-chromosome inactivation [7, 27-32]. The majority of BRCA1-associated cancers are also triple-negative (ER, PR and HER2 negative), express basal cytokeratins and other markers commonly seen in basal-like breast cancers (e.g. p53, P-cadherin and EGFR) [7]. Gene expression profiling also demonstrated that BRCA1-associated cancers segregate strongly with basal-like breast cancers [2, 29, 33, 34]. Although BRCA1 somatic gene mutations are uncommon in sporadic basal-like cancers, these tumors have been shown to have a dysfunction al BRCA1 pathway due to BRCA1 gene promoter methylation and/or BRCA1 pathway transcriptional inactivation [7, 25, 26]. The fundamental biological similarities between hereditary BRCA1-related breast cancers and basal-like cancers suggest that strategies targeting the dysfunctional BRCA1 pathway may be effective in basal-like breast cancers. There is increasing evidence that the DNA repair defects characteristic of BRCA1 related cancers, especially defective homologous recombination, confer sensitivity to certain systemic agents, such as platinum-based chemotherapy and poly(ADP-ribose) polymerase (PARP) inhibitors [34-38]. Indeed, recent clinical studies revealed that sporadic basal-like cancers responded to platinum-based chemotherapy and were associated with a high rate of complete pathologic response [9, 11, 38]. Consistent with the clinical data, our in vitro study also reveals that basal-like breast cancer cells confer specific sensitivity to cisplatin as compared to other chemotherapeutic agents (e.g. doxorubicin or paclitaxel) (Figure 1), further support research into the utility of platinum-ba sed agents in basal-like breast cancers. Given the high specificity and response rate of basal-like breast cancers toward platinum-based therapy, our focus is to maximize the benefits associated with this therapy through drug combination strategies. Using a small chemical library consisted of 160 well-validated and specific inhibitors that target the human kinome, we have identified 6 compounds that significantly potentiate the antiproliferative effects of cisplatin in basal-like breast cancer cells. These compounds include rapamycin, LY364947, AG1478, BAY11-7082, LY294002 and SB203580 which targets the mTOR, TGFà ¯Ã‚ Ã‚ ¢RI, NFà ¯Ã‚ Ã‚ «B, PI3K/AKT and MAPK pathway respectively. Of note, these pathways have been reported previously to be over activated in basal-like breast cancers [7, 17, 18, 20]. To further investigate the mode of interaction between these compounds and cisplatin, we performed a drug combination study using the isobologram approach as described previously (Ref). Out of the 6 compounds identified, rapamycin showed the strongest synergistic effects with cisplatin while others (LY364947, AG 1478, BAY11-7082, LY294002 and SB203580) showed mainly additive effects. This result is consistent with other studies which show that inhibition of mTOR by RNAi or small molecules (e.g. rapamycin, CCI-779, RAD001) enhances cisplatin chemosensitivity in ovarian [39-41], endometrial [42], head and neck [43, 44], lung [45], skin [46, 47] and liver [48] cancers. We next compared the synergistic effects of rapamycin in combination with cisplatin in a panel of luminal-like and basal-like breast cancer cell lines that has been previously validated by gene profiling [15]. Intriguingly, the synergistic effects were observed only in MDA-MB-468, MDA-MB-231 and HCC1937 basal-like cells, but not in MCF-7 or T47D luminal-like cells. Several models have been proposed to explain the synergistic effects of rapamycin and cisplatin in cancer cells. Beuvnk et al., 2005 showed that RAD001 (Everolimus), a rapamycin derivative, dramatically enhances cisplatin-induced apoptosis in wild-type p53 but not mutant p53 tumor cells by inhibiting p53-induced p21 expression [49]. Wangpaichitr et al., 2008 demonstrated that inhibition of mTOR by CCI-779 decreased levels of the anti-apoptotic proteins, BCL2/BCLxL, and increasing apoptosis in lung cancer cells that is resistance to cisplatin [50]. Although these models provide important evidence for mTOR inhibition and cisplatin synergism in cancer cells, it fails to explain the specific synergism we observed in basal-like breast cancer cells, as the basal-like cells that we tested are p53 mutated and do not express high level of BCL2/BCLxL (data not shown). This led us to postulate that a common signal transduction pathway inhibited by rapamycin may be an important component that sy nergizes cisplatin sensitivity in basal-like cells. Since p73 has been reported to mediate cisplatin sensitivity in a subset of triple-negative breast cancer cells [4] and that inhibition of mTOR by rapamycin or RNAi lead to upregulation of p73 [21], we postulated that activation of the p73 pathways might be important for the synergistic effects of rapamycin. To test the role of p73 in rapamycin and cisplatin synergism, we first evaluated the expression of p73 mRNA and protein levels following treatment with cisplatin or rapamycin alone or in combination in MDA-MB-231 cells. Consistent with previous studies, treatment of cells with cisplatin or rapamycin alone induces p73 protein expression followed by transcriptional activation of the 2 potent pro-apoptotic p73 target genes, PUMA and NOXA. When MDA-MB-231 cells were co-treated with rapamycin and cisplatin, the elevation of p73 and its pro-apoptotic target genes were synergistically enhanced. The observed changes in p73 protein in MDA-MB-231 cells, however, were not due to parallel changes in p73 RNA levels, suggesting that inhibition of mTOR might lead to inactivation of a yet unknown p73 specific protein degradation pathway. To validate that the rapamycin and cisplatin synergism is mediated by p73, we generated isogenic MDA-MB-231 and MDA-MB-468 cells that were depleted for p73 using a lentiviral-shRNA that target specifically the transactivating isoform of p73 (TAp73). Indeed, depletion of TAp73 in MDA-MB-231 and MDA-MB-468 cells completely abrogated the synergistic effects of rapamycin suggesting that the synergism between rapamycin and cisplatin required p73 function. Although the combination of cisplatin and rapamycin has not been previously investigated in clinical study, it is worth noting that a phase II neo-adjuvant clinical trial of cisplatin and RAD001 (Everolimus), in patients with triple-negative breast cancers has recently open for recruitment (ClinicalTrials.gov Identifier: NCT00930930), and will be able to address the potential of cisplatin and mTOR inhibitors combination therapy directly. It would be equally intriguing to determine the role of p73 related pathway as potential biomarkers that might predict response to treatment given the pivotal role of p73 in the synergistic effects of mTOR inhibition and cisplatin sensitivity. In conclusion, combination of mTOR inhibitors and cisplatin may be a useful therapeutic strategy in basal-like breast cancers.

John Carlos Story Essay -- Sports

The â€Å"John Carlos Story† is a book about the struggles of growing up black in America at a time when much of the nation was still segregated. John Carlos was a member of the â€Å"Olympic Project for Human Rights.† After winning the bronze medal, John Carlos and a friend and teammate, Tommy Smith, who won the gold medal, raised their fists in opposition of racial inequality and in unity of civil rights. This book shows examples of sociology in everyday life which can be explained through theories and concepts, which centers mainly on the event surrounding their win at the Olympic Games. A seemingly non-emotional, non-verbal, non-violent protest over racism was met with a great deal of negativity at the Olympic Games. Because of the location of their â€Å"protest† over racial inequality, many people did not agree with what they were doing. It was not considered socially acceptable to let their opinion on the matters of race play a part of the â€Å"opinion-free† Olympic Games. Instantly, the audience at the game did not like their way of letting the people know of their beliefs as the crowd went silent. As John Carlos stated in his book, â€Å"There’s something awful about hearing fifty thousand people go silent, like being in the eye of a hurricane. Then, as the national anthem played in full force, the calm before the storm ended and the ‘boos’ started coming down† (Carlos 121). Smith and Carlos were then able to walk off the field but it just got worse, â€Å"the shock was gone and it was officially getting ugly† (Carlos 121). The audience started to yell at them and called them â€Å"anti-American† (Carlos 121). Because of the social setting, people did not understand the purpose of what they were doing or what it had represented. There was ta... ... period of time (1968) in which there was no race cohesion and blacks were not viewed positively by some Americans. Also, the place was likely not the proper location for such a â€Å"demonstration.† Again, people had different reactions and to some, Carlos and Smith were heroes. This can be considered â€Å"labeling theory† which focuses on â€Å"how people define deviance – what is or is not ‘normal’-which is a core issue† (Ballantine/Roberts 175). In the end, those that thought he was a hero were helpful to John Carlos and with his book, society accepts him for his efforts and if there are individuals that still do not, they are the minority in society. Works Cited Ballantine, Jeanne and Roberts, Keith. Our Social World, Introduction to Sociology. Pine Forge Press, 2011. Carlos, John. The John Carlos Story. (With Dave Zirin). Haymarket Books, Chicago, IL 2011.

Mus 100 Study Guide

MUS 100 FINAL STUDY GUIDE CHAPTER 17: – Fortepiano: early piano, named for its range of dynamic levels; it was smaller and less sonorous than the modern instrument. – Classical style: restrained, objective style of art. Classical refers to Western music characteristic of the period from 1750-1825. Composers: – Mozart: Invested much of his music with a degree of emotion expression unusual for his time. Never allowed emotion to dominate his art. – Haydn: Wrote pleasant, good-natured music throughout his long life. Wrote masses, oratorios, and other religious compositions for church and for concert performance. Beethoven: Wrote masses, oratorios, and other religious compositions for church and for concert performance. CHAPTER 18: – Form: organization and design of a composition, or of one movement within a composition. – Symphony: multimovement orchestral form. – Sonata-Allegro: â€Å"first movement form†. The 3 sections: expositio n, development, and recapitulation-form a binary design. – Exposition: first section of a fugue or of a sonata-allegro. – Development: 2nd section of the sonata-allegro; it moves through many keys. – Recapitulation: 3rd section of the sonata-allegro.Reviews the material of the exposition, presenting it in a new light. – Coda: Meaning, â€Å"tail†; a closing section. – Minuet and Trio: ABA. Often the 3rd movement of a symphony, sonata, or string quartet. Consists of two minuets, the second (trio) lighter and more lyrical than the first. – Cadenza: extended passage for solo instrument; typical feature of a solo concerto. – Rondo: ABACA. Form in which various episodes alternate with the opening material. The tempo is usually fast, and the mood merry. – String Quartet: chamber ensemble consisting of two violins, a viola, and a cello. Sonata (classical period): a multimovement composition for one or two solo instruments. CHAP TER 19: – Overture: introductory orchestral piece. – Comic Opera (ope’ra comique, singspiel, opera buffa): Operas light in mood, modest in performing requirements, written in the vernacular language of the intended audience. – Requiem: mass for the dead. – Ensemble Finale: final scene of a musical show in which several soloists simultaneously express, in different words and music, their individual points of view. CHAPTER 20: – Motive: short melodic phrase that may be effectively developed. Art song: concert setting of a poem, usually by a well-known poet, to music. – Lieder: German art songs. – Song cycle: sets of songs by one composer, often using texts all by the same poet. Composers: – Schubert: earliest master of romantic art son. Composed 143 songs at 18. â€Å"Godfather† of the romantic period genre. CHAPTER 21-22: – Cyclic form: multimovement form unified by recurrence of the same or similar melodic material in two or more movements. – Absolute music: instrumental music having no tended association with a story, poem, idea or scene; non-program music. Concert overture: one movement orchestral composition, often inspired by literature and dramatic in expression, yet generally subject to analysis according to classical principles of form. – Program symphony: symphony (composition for orchestra in several movements) related to a story, idea, or scene, in which each movement usually has a descriptive title. – Idee fixe: single melody used in several movements of a long work to represent a recurring idea. – Thematic transformation: variation of thematic or melodic material for programmatic purposes.Sometimes called metamorphosis. – Dies irae: Gregorian chant for the dead. – Symphonic poem (tone poem): programmatic composition for orchestra in one movement, which may have a traditional form (such as sonata/rondo) or an original irregular form . Composers: – Brahms: misplaced classicist. Poured the warmest Romantic emotional content into his classical forms. He based his music on models from the past. – Berlioz: his works were based on unrequited love. Used the idee fixe, which was a melodic reference to his beloved. CHAPTER 23: Character piece: relatively short piano piece in a characteristic style or mood. – Nocturnes: Piece expressing the â€Å"character† of night. – Prelude: short independent or introductory piece for keyboard. – Etude: a virtuosic instrumental study or â€Å"exercise† intended for concert performance. – Rubato: romantic technique of â€Å"robbing† from the tempo at some points and â€Å"paying back† at others. Composers: – Chopin: only great composer who wrote almost exclusively for piano. Most pieces are miniatures. Virtuoso pianist, most famous for lyrical and melancholic melodies.CHAPTER 25: -Post-romanticism: general ter m for several romantic styles that succeeded the dominance of German Romanticism and preceded the return of classicism to the arts. – Atonality: avoidance of a tonic note and of tonal relationships in music. – Impressionism: style of painting and music that avoids explicit statement, instead emphasizing suggestion and atmosphere. – Primitivism: style inspired by primitive works of art and by the relaxed life of unsophisticated cultures. – Pizzicato: technique of plucking string instruments.Composers: – Mahler: post-romantics. Wrestled with conflicting romantic and classical ideals. – Strauss: leader of post-romantic composers. Strictly classical style but developed romantic techniques. – Debussy: first musician labeled an impressionist. Developed unusual harmonies and exotic timbres. – Schoenberg: inventor of the 12-tone method (serialism) > Using the 12 pitches equally. > 12 tone row: playing the 12 pitches in whatever order; no repeated tones until the row has been fully played. > Wrote in a free atonal style gt; Drifted away from traditional harmony and experimented other styles – Stravinsky: went through an early ballet period before the war. He went through a neo-classical period. > Primitivism: movement in the second decade of the 20th century. Reveals romanticism characteristics. Characterized by strong savage rhythms, dissonant combinations of sound and narrow melodies. > â€Å"Rite of Spring†: controversial piece, ballet, and scandal piece CHAPTER 27: – Experimentalism: exploration of previously unknown aspects of musical sound. Polytonality: two or more keys at the same time. – Tone cluster: chord built on seconds. – Prepared piano: piano whose timbre and pitches have been altered by the application of foreign materials on or between the strings. – Twelve-tone technique: arrangement of the twelve chromatic pitches into a tow that provides the melodic and harmonic basis for a music composition. Row: series of tones on which a serial composition is based. Composers: – Schoenberg: inventor of the 12-tone method (serialism) – Weberm: developed his own styles: lean, clean, delicate, and strong. Ives: invented polytonality (incorporating of two different keys). – Cowell: invented the plucking of a piano sound. – Cage: 1912-1992 not trained as a musician. Brought up in Los Angeles. Became a composer. > Alatoric: predetermined sounds and just guessed when it should be played. > Conceptual art: piece called 4 minutes a 33 seconds – just the sounds in CHAPTER 28: – Neoclassicism: 12th century version of classicism in music. – Neoromanticism: 12th century version of a romantic approach to music. –Minimalism: style of music based on many repetitions of simple melodic lines that gradually change and slowly evolve patterns and rhythmic patterns. Composers: – Copland: American nationa list composer > â€Å"Dean of American Music† – Gershwin: Best known of all American opera, filled with the characteristic sounds of jazz, including syncopated rhythms, expressive vocal catches and slides. – Prokofiev: focused on neoclassical music. – Barber: focused on neoromanticism. > Adagio for string orchestra (tonal piece) – Reich: focused on minimalism. – Glass: focused on minimalism.

Hawaiian Punch Essay

In July 2004, Kate Hoedebeck was promoted to director of marketing for the Hawaiian Punch brand. The CEO of the parent company, Cadbury Schweppes Americas Beverages, set a goal to create a premier marketing and sales organization that capitalizes on the terrific brand recognition. To reach this goal each brand is required to meet the desires of their customers; bottlers, distributors, retailers, and consumers. When creating the 2005 business marketing plan Kate faced a problem. The Hawaiian Punch brand had two manufacturing, sales, and distrution networks. Her objective was to determine the role each network played in the future sales and profitability of the company. III.Alternate Solutions a. Product Positioning Hawaiian Punch needs to clairify the positioning of its brand. The positioing statement needs to clairify its target markets, and focus on the values of its customers. The majority of the Hawaiian Punch juice drinkers ranged from 6 – 18 years of age. Adolescences purchase the drink most frequently from the soda aisle. Of cosnumers aged 6-12, the majority of the purchases were made in the juice aisle by parental/guardian figures. Including the fun consumption experience, its unique taste, and high viatim C content is the posotion the brand wants to own. To out perform the competitors it is necessary to establish these elements and clearly define where Hawaiian Punch sits. Its more than your normal juice, but healthier than soda. b. Innovations Hawaiian Punch would benefit from launching new sizes and flavors. Retailers believed the potential of market growth hinged on creating new packing and flavors. Hawaiian Punch recently experienced slowed growth due to the decrease in the juice market. Hawaiian Punch maintained market share, and Trade Customers believed the market would rebound. To take advantage of the market portential, Hawaiian Punch should innovate its product size offering. Currently Hawaiian Punch is packaged in 1-gallon bottle, a half-gallon bottle, a 2-liter bottle, a 20-ounce bottle, a 6.75-ounce single serve standup pouch, and 12-ounce cans. According to customer insights, 77% of Hawaiian Punch buyers purchased only size. According to reasearch, 68% of buyers that purchaed the 1-gallon bottle and 53% of buyers that purchased  the half-gallon size were exclusive in their size purchases. The 2-liter and 20-ounce bottle purchasers experienced the least amount of size loyalty. Despite having customer enjoying the different size offerings the numerous amount of sizes caused distribution constraints and shelving issues at numerous locations. By eliminating certain size offerings with low size loyalty and creating new ones while maintaining variety will elliviate contraints and provide new opportunties for customers. The brand would also benefit from new flavor innovation. Hawaiian Punch currently has 11 product flavor offerings. The most popular and original flavor maintains to be fruit Punch. Out of the 11 product offerings there has been a wide variety of success. Hawaiian Punch needs to establish stable products in both their Finished goods and Direct-Store Delivery networks. Products that do not show profitabilty should be elimiated. Introducing new flavors to the Direct-Store Delivery will benefit because of the large market potential and smaller size offerings. c. Allowances and Advertising Hawaiian Punch should increase its advertising budget and allowances for marketing. Currently Hawaiian Punch spends $2.2 Million on advertising a year. This accounts for 1% of advertising by Hawaiian Punch’s competitve juice brands collectively. Their current advertising mix consists of only radio and magazine print ads. The largest amount of advertising dollars spent is in television which currently Hawaiian Punch does not participate. This media outlet accounts for roughly 82% of total media dollars by competitors. Increasing media dollars will help reach new customers, and enforce new flavors to the brand. Of the advertising dollars spent much should be included in recreating the â€Å"Punchy’ Image. Past years have downplayed this aspect of the product brand, but still plays a large part in its recognition. By including Punchy in their advertising they should focus their attention on children between 8-18 years old. Punchys image will play a large role in delivery the benefits of the fruit drink and the position in the market. IV.Selected Solution It is recommended that Hawaiian Punch position their brand and maximize profit by increasing advertising and innovating their products. Having Dual distribution networks it is necessary to establish market segments. The  Finished Goods Network buyer is mainly mothers/guardians. This segment presents the largest amount of repurchasing and brand loyalty. Focusing advertising on the energic times and healthy benefits will persuade mothers ino purchasing the drink because it offers V.Conclusions Entering into a licensing agreement is not a good decision in the European market, the three year agreement is too short and more importantly Baxton is allowing too much of the revenue to go to Bar Maisse, they need an arrangement where they receive a larger portion of the proceeds. At the same time, choosing to enter the European market is too risky and has too many unknowns. The expertise of Bar Maisse is needed for success to be likely; otherwise there are too many unknowns with where operations should be based on what markets are more likely to yield successful sales. While focusing on Europe and entering the market at this time does appear to be the correct decision, it should not be done at the expense of increasing growth in the United States. That market is under developed and is not being capitalized, as it should be.

Individualism

Individualism 1) An individualist is considered to be someone with personality and character, someone who is not easily intimidated by social pressure or customs, someone with a personal opinion and a singular view of the world. Because modern society finds it important that people think independently, decide autonomously and take personal initiatives, the concept of individualism has acquired a positive connotation. However, individualism is also linked with the tendency to withdraw from social life and turn in towards oneself. 2) Which drives people to withdraw into a small, enclosed world consisting of their family and a few select friends, leaving the rest of society to its own devices. 3) Courageous individualism refers to the dedication shown by people who have independently chosen for something or someone and work energetically for its sake in spite of opposition from family or society; indifferent individualism refers to people who have either lost all hope in their fellow human beings or are no longer interested in initiatives of a social nature, with the result that they are prepared to withdraw from society at all costs.Individualism4) people who have received less education and who find themselves in a precarious economic situation are more quickly inclined to believe that it is 'everyone for himself' in our society: on this view, politicians pay no attention to the needs of the people, society develops in a chaotic and unpredictable manner, the welfare we once enjoyed belongs definitively to the past, life has little meaning, and there are no longer any people or associations to which one can make an appeal. 5)On the other hand, whoever considers work not only as one's own achievement but also as an opportunity provided by the community, and whoever sees it as one's duty as a citizen to ensure opportunities for fellow citizens...

Implementation and evaluation plan Assignment Example | Topics and Well Written Essays - 1000 words

Implementation and evaluation plan - Assignment Example The evidences focus on the systematic reviews of the recommendation and literature according to evidence level, using criteria that are pre-defined. The treatment lines are flagged out based on methods that included expert clinical support and evidence. Interpersonal Therapy (IPT) and Cognitive-Behavioral Therapy (CBT) have the most evidence efficiently. Additionally, both maintenance and acute MDD phases, and have undergone study in combination with the anti-depressants. Cognitive-Behavioral Therapy is well researched in connection with bibliography and Computer-Developed methods. The evidence of Cognitive-Behavioral Analysis Systems of Psychotherapy and Behavioral Activation are significant. However, the evidences need replication. Modern psychotherapies that include Commitment Therapy and Acceptance, Motivational Interviewing, and Mind-fullness-Based Cognitive therapy have no relevant evidence as psychodynamics therapy and acute treatments. Although various forms of psychotherapy have been studied, only a few types have been subjected to evaluation in RCTs. Evidence of combination of various kinds of antidepressant and psychotherapy medication is confined despite the vast use of the therapies concomitantly. Lamotrigine is appropriate to the client's problem because according to RCT, almost half of the patients responded positively. This is two times experienced with the placebo group according to level 1 (22, 23). Additionally, recent RCT in clients who responded positively on bipolar depression under lithium revealed that Lamotrigine add-on is superior compared to placebo add-on when used to treat depressive symptoms. This is evident by greater rise in the MADRS scores. According to large RCT, combining olanzapine and fluoxetine is slightly but significantly better compared to lamotrigine immunotherapy. However, Lamotrigine is highly tolerated. Combination of fluoxetine and Olanzapine was linked with statistical significance and positive results in maniac a nd depressive systems when compared to Lamotrigine. On the other hand, the size of its effect was small, and no difference was recorded in their response rate. According to the guidelines of 2005 bipolar, only a single RCT had shown antidepressant efficiency of quetiapine monotherapy in treating bipolar depression (25). Its rate of remission was 52.9 percent in the group consuming 600mg per day and 300 mg per day of quetiapine relative to 28.4% for placebo (Sagar et al., 2009). At earlier stages of testing, quetiapine monotherapy was thought to be immature as an antipsychotic depressant for bipolar. The second RCT confirmed the efficiency of quetiapine monotherapy (26). The study recommended quetiapine monotherapy as bipolar depression first-line option. Further evaluations (25) showed an improvement in health-related standard of life. Client's input can be obtained by conducting a randomized controlled trial. This involves assessing a period's systematic intervention program. This includes carrying out a psycho education program of structured group, monthly telephone evaluation and monitoring of medication adherence and mood symptoms, feedback during the treatment of mental health providers. Additionally, facilitate proper follow-up care and as-required crisis and outreach intervention. The patients can be monitored by augmenting their self-management skill through psycho education. Consequently, the decision support of the provider by

Math Edu Essay Example | Topics and Well Written Essays - 500 words

Math Edu - Essay Example First, students can be shown a square along with the length of its sides. Ask students to calculate its area. Next, pass out several cubes to the class so that there are enough for everyone to look at. These cubes should preferably have the same approximate side measurements as the square previously displayed. After explaining that surface area is the total area of the surface on the outside of an object, ask students how many sides there are, and what the shape of each side is. Ask them to make the connection themselves as to how to find the surface area of the cube. If they do not get it at first, hold up a picture of the square they just found the area of and compare it to the sides of the cube. They should be able to recognize that to find the surface area of the cube you multiply the area of the square by the number of squares/sides on the cube. Another good introduction or way to reiterate an understanding of how to find the surface area of a cube is to transform a two-dimensional drawing into a three-dimensional object. Pass out pieces of paper with six large squares on them. These squares should be arranged in a way that four are next to each vertically and one square is on each horizontal side of the third square. (It should resemble a cross).