Improved Glottal Closure Instant Detector Based On Linear Prediction And Standard Pitch Con
是智能科技让我们变笨了吗英语作文

是智能科技让我们变笨了吗英语作文全文共3篇示例,供读者参考篇1Have Intelligent Technologies Made Us Dumber?As I sit here typing away on my laptop, with multiple tabs open for research, music streaming in the background, and my phone buzzing with notifications every few minutes, I can't help but wonder – has all this intelligent technology actually made me dumber? It's a question that's been on my mind a lot lately, especially as I see my generation and those younger than us becoming increasingly reliant on our devices and the virtual world.On one hand, I have the entirety of human knowledge quite literally at my fingertips. Need to know the capital of Uzbekistan? Just Google it. Stuck on a complicated math problem? There's a website or app for that. Curious about the plot of that new movie everyone's talking about? I can read online reviews and synopses in seconds. With so much information so readily available, it seems like we should be getting smarter and more knowledgeable by the day.But then I look around at my classmates buried in their phones, unable to concentrate for more than a few minutes at a time without checking Instagram or Snapchat. I've noticed my own attention span has gotten incrementally worse over the years too. Reading physical books and even lengthy articles has become a struggle when I'm so accustomed to the bite-sized information and constant stimulation of the internet and social media. Critical thinking and analysis seem to be falling by the wayside as we rely more on skimming and searching for quick answers online.Then there's the fact that we're outsourcing huge portions of our brain's capabilities to our devices. Why bother committing facts to memory or working through complex math when I can just ask Siri or Alexa? Your phone has a calculator and maps app – so do you really need to learn arithmetic or get good at reading maps? With so much knowledge externalized and stored in silicon rather than grey matter, our own cognitive abilities could potentially atrophy over time.On the flip side, you could argue that intelligent technologies have simply shifted what skills are most valuable in the modern world. Maybe rote memorization and arithmetic aren't as crucial when you have powerful computers to assistwith those tasks. Instead, the cognitive skills of the future could lie in areas like problem-solving, critical thinking, creativity, and learning how to learn. If that's the case, then perhaps these technologies are pushing the evolution of human intelligence down a different path rather than making us universally dumber.There's also the question of access to education to consider. In many developing parts of the world, people finally have access to vast educational resources online that were previously unattainable. Intelligent technologies have opened up new frontiers of learning for billions across the globe. Even in more prosperous nations, these tools have helped level the playing field for students in underprivileged school districts or those with learning disabilities. From interactive teaching apps to online tutors to text-to-speech capabilities, the same innovations that may be inhibiting critical thinking for some are expanding minds and creating opportunities in other contexts.So does the good outweigh the potential negatives? It's hard to say for certain, as we're still in the relatively early days of widespread intelligent technology adoption. As with most transformative technologies throughout human history, there are bound to be ranges of benefits and drawbacks that will varybased on individual circumstances and how these tools are implemented and used.My personal opinion is that intelligent technologies haven't made us broadly dumber yet, but there are certainly concerning trends and risks that we need to be cognizant of, especially when it comes to younger generations. We have to be intentional about using these tools in ways that augment and amplify our natural intelligence rather than replace or diminish it.Perhaps curricula and teaching methods need to adapt to put more emphasis on higher-order cognitive skills and intellectual discipline in the digital age. Or we could look for ways to build more mindfulness into our relationships with technology to prevent unhealthy overreliance. But simply sticking our heads in the sand or trying to eschew these powerful technologies altogether doesn't seem like a pragmatic solution in our rapidly evolving world.Ultimately, the answers may lie in striking a balance – using intelligent technologies as complements to our own human strengths and weaknesses, while remaining self-aware enough to recognize where we're diminishing our intellectual capacity rather than enhancing it. As long as we can think critically about our relationship with technology and judiciously determinewhich tasks are better suited for human brains versus artificial intelligence, perhaps we can enjoy the best of both worlds.After all, the human mind is an incredible thing – a cognitive marvel that has created ingenious tools and technologies, composed sublime art and music, formulated insights into the nature of our reality, and launched exploratory voyages across this planet and beyond. While our intelligent technologies are impressive, they are still fundamentally limited by the creativity, imagination, and problem-solving abilities of the human minds that conceived of and constructed them in the first place. Let's not be so quick to discount our own intelligence and capabilities, even as we look to augment them with increasingly powerful technologies.篇2Has Technology Made Us Dumber?It's no secret that technology has become deeply intertwined with nearly every aspect of our lives. We're constantly surrounded by smartphones, laptops, tablets and other devices that provide us with instantaneous access to an endless stream of information and entertainment at our fingertips. But while technology has made many tasks moreefficient and convenient, there's an ongoing debate about whether our reliance on these digital tools is actually making us less intelligent.On one hand, it's easy to see how technology could be perceived as a crutch that diminishes our critical thinking and problem-solving abilities. With services like Google and Wikipedia just a click away, we no longer have to commit vast amounts of information to memory the way previous generations did. And with spell-check, autocorrect and predictive text features, we may be losing our grasp on proper spelling, grammar and written communication skills.There's also the argument that technology creates a continuous state of distraction and overstimulation that hampers our ability to focus deeply on any one task. With notifications constantly buzzing on our devices, enticing us to shift our attention elsewhere, some claim that we're developing shorter attention spans and struggling to think critically about complex issues.However, I would argue that while technology presents some potential drawbacks, it has ultimately expanded our intellectual capabilities in numerous ways and made us smarteras a society. Rather than making us dumber, I believe technology has simply changed the way we think and acquire knowledge.One major advantage of technology is that it has exponentially increased the amount of information that's readily available to us. A few decades ago, learning was largely limited to what could be found in physical libraries and textbooks. But today, we have the entirety of human knowledge quite literally at our fingertips through tools like online databases, ebooks, research journals and learning platforms like Coursera and edX.While critics claim this makes us overly reliant on looking up information rather than learning it ourselves, I would counter that having such a wealth of knowledge so easily accessible actually augments our ability to learn new concepts more quickly. We no longer have to waste time memorizing dry facts when we can simply look up data as needed, freeing up cognitive resources to focus on deeper understanding, creative thinking and synthesizing information in meaningful ways.Technology has also opened up incredible opportunities for interactive, experiential learning that simply weren't possible in the past. Between virtual reality environments that allow us to vividly explore ancient civilizations and distant galaxies, simulation software that lets us run experiments risk-free, andeven video games that require strategic planning and problem-solving, we now have engaging, hands-on methods to learn that go far beyond just reading from textbooks.And despite claims that we're becoming hopelessly distracted, certain technologies can actually help us focus better than ever before. Noise-cancelling headphones enable us to block out external sounds that could divert our attention. Apps like Forest allow us to "plant trees" while we work, gamifying the process of avoiding technological distractions. Even the very smartphones that are often denounced as disruptive have settings like Do Not Disturb that create distraction-free workspaces when we need to concentrate.Moreover, technology has dramatically improved accessibility when it comes to education. Students who may have struggled in traditional classroom environments can now take advantage of personalized learning tools that cater to their unique needs and allow them to learn at their own pace. Those with disabilities such as vision or hearing impairments can utilize assistive technologies that give them more independence. And for the millions around the world who lack access to quality schools, the internet grants them the ability to tap into anear-limitless supply of free or low-cost educational resources.In terms of intelligence itself, while technology may seem to diminish certain skills, it has also nurtured different types of cognitive abilities that will be essential for succeeding in the 21st century workplace. With so much information constantly inundating us, we're getting better at filtering out irrelevant data to identify what's truly important. Tech has pushed us to become more adept at multimedia learning, absorbing and synthesizing information from a diverse array of sources like videos, podcasts, graphics and more. And our frequent use of search engines and databases has sharpened our talent for quickly finding the specific knowledge we need for any given situation.Ultimately, I believe this cognitive outsourcing and development of new skills represents an overall net positive when it comes to human intelligence. While technologies may reduce our reliance on rote memorization, they have opened up opportunities to nurture deeper cognitive and creative abilities that will prove far more valuable as we move into an era where innovation and critical thinking are paramount.Of course, as with anything, maintaining a balanced approach with technology is crucial. We can certainly become too dependent on these tools and allow our brains to atrophy if we're not self-aware and don't challenge ourselves occasionally.Reading physical books, handwriting notes, and taking intentional breaks from screen time are all healthy habits that should be preserved. But overall, when used responsibly as a complementary aid, I believe technology has made us smarter by evolving the way we learn, expanding our access to knowledge, and cultivating new cognitive skills that will serve us well in our rapidly changing world.篇3Has Smart Technology Made Us Dumber?As I scroll mindlessly through my social media feeds, I can't help but ponder the impact that modern technology has had on our cognitive abilities. With information quite literally at our fingertips, it's hard not to wonder if we've become overly reliant on these digital crutches, potentially stunting our intellectual growth in the process.On the one hand, the technological advancements of the 21st century have revolutionized the way we access and process information. Gone are the days of scouring through dusty encyclopedias or waiting for the evening news to stay informed. Today, a simple voice command or a few taps on a screen canunlock a vast repository of knowledge, spanning virtually every topic imaginable.This unprecedented access to information has undoubtedly broadened our horizons and facilitated a more well-rounded understanding of the world around us. However, as convenient as these digital assistants and search engines may be, there's a growing concern that they've inadvertently diminished our ability – and perhaps even our inclination – to think critically and retain knowledge.Think about it: how many times have you found yourself relying on your smartphone to settle a friendly debate or looked up a simple fact, only to forget it moments later? It's as if our brains have become accustomed to outsourcing the task of remembering to our trusty devices, leading to what some experts have dubbed the "Google effect" – a phenomenon where we tend to forget information that's readily available online.But it's not just our memory that's at risk of atrophy; our problem-solving skills and attention spans are also under siege. With virtually everything we could ever want or need just a tap away, it's becoming increasingly challenging to stay focused and engaged. We've grown accustomed to constant stimulation andinstant gratification, making it harder to tackle complex tasks that require sustained concentration and deep thinking.And let's not forget about the impact of social media and its relentless stream of bite-sized content. While platforms like TikTok and Instagram have undoubtedly revolutionized the way we consume and share information, they've also contributed to a sort of cultural ADD, where our attention spans are constantly fragmented by an endless barrage of memes, videos, and status updates.But perhaps the most insidious danger of our reliance on smart technology lies in its potential to erode our critical thinking skills. With so much information readily available, it's easy to fall into the trap of accepting whatever we see or read at face value, without questioning the source or scrutinizing the validity of the claims.This lack of skepticism can be particularly problematic in an age where misinformation and fake news run rampant, often masquerading as legitimate sources. If we're not careful, we risk becoming passive consumers of information, rather than active, discerning thinkers.On the flip side, however, there are those who argue that smart technology has actually enhanced our cognitive abilities inprofound ways. By offloading certain mental tasks to our devices, we've been able to free up valuable brain power and redirect our focus toward more complex and creative endeavors.Proponents of this view contend that the sheer volume of information at our disposal has forced us to become more adept at filtering out irrelevant data and synthesizing disparate pieces of knowledge – skills that are arguably more valuable in the information age than mere memorization.Moreover, they argue that the interactive and multimedia-rich nature of modern technology has fostered new forms of learning and engagement, catering to a wider range of learning styles and potentially unlocking untapped cognitive potential.Personally, I find myself torn between these two perspectives. On one hand, I can't deny the conveniences and educational opportunities that smart technology has afforded me. From researching topics for school assignments to staying connected with friends and family across the globe, my life has been immeasurably enriched by the digital age.At the same time, however, I've noticed a concerning trend in my own habits and those of my peers. I've caught myself mindlessly scrolling through social media feeds for hours on end,only to realize that I've retained little to nothing from the content I've consumed. I've also found it increasingly difficult to focus on tasks that require sustained attention, often succumbing to the siren call of my smartphone or the allure of multitasking.So, has smart technology made us dumber? The answer, I suspect, is a resounding "it depends." Like any tool, the impact of technology on our cognitive abilities ultimately lies in how we choose to wield it.If we approach these devices as mere repositories of information, outsourcing our mental faculties to the digital realm, then yes – we risk becoming intellectually complacent and allowing our critical thinking skills to atrophy.However, if we learn to strike a balance, harnessing the power of technology to supplement and enhance our natural abilities, then we may very well find ourselves on the cusp of a cognitive renaissance, where the boundaries of human knowledge and understanding are pushed further than ever before.Ultimately, the onus is on us – as individuals and as a society – to cultivate a healthy relationship with smart technology. We must remain vigilant, questioning the information we consumeand actively exercising our minds through activities that promote critical thinking, problem-solving, and sustained focus.Only then can we truly reap the benefits of the digital age while safeguarding the very faculties that make us uniquely human.。
美FDA批准第一种免疫球蛋白皮下输注剂Hizentra用作原发性免疫缺陷患者替代治疗

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用于哮喘的生长因子治疗[发明专利]
![用于哮喘的生长因子治疗[发明专利]](https://img.taocdn.com/s3/m/1a3b73119e31433238689337.png)
专利名称:用于哮喘的生长因子治疗
专利类型:发明专利
发明人:唐娜·伊丽莎白·戴维斯,史蒂芬·T·霍尔盖特,林恩塞·M·汉密尔顿,萨拉·玛格丽特·普迪库姆,奥德丽·里克
特
申请号:CN200580038645.3
申请日:20050913
公开号:CN101056649A
公开日:
20071017
专利内容由知识产权出版社提供
摘要:本发明涉及对于治疗或防止哮喘患者的10种支气管上皮损害的生长因子的用途,所述生长因子是表皮生长因子(EGF)类似物、KGF或KGF类似物。
用于该目的的适合的EGF类似物靶向EGF受体并在哮喘患者中显示与气道成纤维细胞相比促进支气管上皮细胞优先增殖的能力。
申请人:南安普敦大学
地址:英国南安普敦
国籍:GB
代理机构:中科专利商标代理有限责任公司
代理人:王旭
更多信息请下载全文后查看。
提高了接触轨取流质量 英文

提高了接触轨取流质量英文Improving Contact Lens Fluid QualityThe quality of contact lens fluid is a crucial factor in maintaining the health and comfort of the eyes for individuals who wear contact lenses. Contact lens fluid, also known as contact lens solution or care solution, is a liquid used to clean, disinfect, and store contact lenses. It plays a vital role in ensuring the lenses are properly cared for and free of any harmful bacteria or debris that could potentially cause eye irritation or infection.One of the primary functions of contact lens fluid is to clean the lenses. The fluid contains surfactants and other cleaning agents that help remove any buildup of proteins, lipids, or other deposits that may accumulate on the lens surface during wear. This cleaning process is essential to prevent the lenses from becoming cloudy or uncomfortable, which can lead to reduced visual acuity and increased risk of eye health issues.In addition to cleaning, contact lens fluid also plays a crucial role in disinfecting the lenses. The fluid typically contains antimicrobial agents, such as hydrogen peroxide or certain types of preservatives,that help kill any harmful bacteria or microorganisms that may be present on the lens surface. This disinfection process is critical to prevent the development of eye infections, such as bacterial keratitis or acanthamoeba keratitis, which can have serious consequences if left untreated.Furthermore, contact lens fluid is responsible for maintaining the proper storage and hydration of the lenses when they are not in use. The fluid helps to keep the lenses moist and prevent them from drying out, which can lead to increased discomfort and potential damage to the lens material. Proper storage and hydration are essential for ensuring the lenses remain in good condition and can be safely reused.Given the importance of contact lens fluid in maintaining eye health and comfort, it is crucial that the quality of these solutions is continuously improved and optimized. One way to achieve this is through ongoing research and development efforts focused on enhancing the formulation and performance of contact lens fluids.One area of focus in improving contact lens fluid quality is the development of more effective cleaning and disinfection agents. Researchers are exploring new compounds and methods that can more efficiently remove stubborn deposits and kill a wider range of microorganisms, without compromising the comfort or safety of thelenses.Another important aspect of improving contact lens fluid quality is the optimization of the fluid's pH and osmolality. These properties can greatly impact the compatibility of the fluid with the eye's natural tear film and the comfort experienced by the wearer. By carefully adjusting the pH and osmolality of the fluid, manufacturers can ensure that it is well-tolerated by the eyes and does not cause any irritation or discomfort.Additionally, the incorporation of advanced preservative systems in contact lens fluids is an area of ongoing research and development. Preservatives play a crucial role in preventing microbial growth and ensuring the long-term stability of the solution, but they must be carefully selected and balanced to minimize any potential adverse effects on the eyes.Furthermore, the packaging and storage of contact lens fluids is also an important consideration in improving their quality. Manufacturers must ensure that the containers used to store the fluids are designed to maintain the integrity and sterility of the solution, preventing any contamination or degradation during the product's shelf life.By addressing these various aspects of contact lens fluid formulation, manufacturers can continuously improve the quality andperformance of these essential products, ultimately enhancing the overall eye health and comfort of contact lens wearers.In conclusion, the quality of contact lens fluid is a critical factor in maintaining the health and comfort of the eyes for individuals who wear contact lenses. Ongoing research and development efforts focused on improving the cleaning and disinfection capabilities, pH and osmolality optimization, preservative systems, and packaging of these solutions are essential to ensure that contact lens wearers can enjoy a safe and comfortable wearing experience.。
八年级科技前沿英语阅读理解25题

八年级科技前沿英语阅读理解25题1<背景文章>Artificial intelligence (AI) has been making remarkable strides in the medical field in recent years. AI - powered systems are being increasingly utilized in various aspects of healthcare, bringing about significant improvements and new possibilities.One of the most prominent applications of AI in medicine is in disease diagnosis. AI algorithms can analyze vast amounts of medical data, such as patient symptoms, medical histories, and test results. For example, deep - learning algorithms can scan X - rays, CT scans, and MRIs to detect early signs of diseases like cancer, pneumonia, or heart diseases. These algorithms can often spot minute details that might be overlooked by human doctors, thus enabling earlier and more accurate diagnoses.In the realm of drug development, AI also plays a crucial role. It can accelerate the process by predicting how different molecules will interact with the human body. AI - based models can sift through thousands of potential drug candidates in a short time, identifying those with the highest probability of success. This not only saves time but also reduces the cost associated with traditional trial - and - error methods in drug research.Medical robots are another area where AI is making an impact.Surgical robots, for instance, can be guided by AI systems to perform complex surgeries with greater precision. These robots can filter out the natural tremors of a surgeon's hand, allowing for more delicate and accurate incisions. Additionally, there are robots designed to assist in patient care, such as those that can help patients with limited mobility to move around or perform simple tasks.However, the application of AI in medicine also faces some challenges. Issues like data privacy, algorithmic bias, and the need for regulatory approval are important considerations. But overall, the potential of AI to transform the medical field is vast and holds great promise for the future of healthcare.1. What is one of the main applications of AI in the medical field according to the article?A. Designing hospital buildings.B. Disease diagnosis.C. Training medical students.D. Managing hospital finances.答案:B。
Technical Guide for Non-Radioactive Nucleic Acid Labeling & Detection_KPL

Technical Guide for Non-Radioactive Nucleic Acid Labeling and Detectionw w w.k p l.c o mTable of Contents Page Chapter 1 – Overview of Non-Radioactive Labeling and Detection3Chapter 2 – Nucleic Acid Probe Labeling 9Chapter 3 – Southern Blotting 23 Chapter 4 – Northern Blotting 33 Chapter 5 – In-Situ Hybridization41 Chapter 6 – Troubleshooting Guide55 Chapter 7 – Appendix65 Miscellaneous Applications65Buffer Recipes68Related Products69 1800-638-3167 • KPL, Inc. • 301-948-77553800-638-3167 • KPL, Inc. • 301-948-7755Non-rad vs. 32PA variety of methods have been developed to detect specific nucleic acid sequences immobilized on membranes (i.e., dot/slot blot, Southern blot,Northern blot, South-Western blot, colony and plaque lifts) and localized in situ in cells and tissues. 32P has traditionally been used due to the intensity of signal it produces and, thus its ability to facilitate the detection of small amounts of biomolecules on blots. However, 32P is not without its shortcomings. These include issues associated with handling and disposing of hazardous material, long exposure times and short half-life, limiting the stability of probes.In recent years, non-radioactive nucleic acid labeling and detection method-ologies have become available in response to a desire by researchers and their institutions to move away from the use of radioisotopes. Advancements made in the areas of chemiluminescence and fluorescence have allowed for an easier transition. In non-radioactive assays, signal is generated through an enzymaticreaction with a chemiluminescent or chromogenic substrate; alternatively ,detection can occur through the appropriate excitation and emission of a fluorophore-labeled probe. For those laboratories seeking replacement technology to 32P without significant investment in instrumentation, chemiluminescent detection enables equivalent results, easily and quickly captured on digital imaging systems or X-ray film shortly after exposure. It is now possible to detect femtogram quantities of nucleic acid in as little as 10 minutes when a hapten and reporter molecule are used in conjunction with a chemiluminescent substrate. The hazards and regulatory issues surrounding 32P-based detection are no longer a trade-off for sensitive, reproducible results.Optimal non-isotopic nucleic acid detection depends primarily on three variables: 1) the molecule or compound used to label the probe,2) hybridization conditions, and 3) the detection method.Figure 1: Comparison of Southern Blot Detection using Detector Labeling and Detection vs. 32P . Detection of single copy gene, n-myc, from 5 µg of human genomic DNA. Panel 1A: Blot detected with a biotinylated n-myc probe and Detector AP Chemiluminescent Blotting Kit in a 10 minute film exposure. Panel 1B: Detection of the same gene with 32P-labeled probe, 16 hour exposure.Figure 2: Comparison of Southern Blot Detection using Detector Labeling and Detection vs. 32P . HeLa cell total RNA detected using an in vitro transcribed biotinylated ß-actin RNA probe and the Detector AP Chemiluminescent Blotting Kit in a 10 minute film exposure (2A) and a 32P-labeled probe in a 3.5 hour film exposure (2B).Component Effect ActionSodium ion concentration Favors formation of hydrogen bonds Increasing [Na+] increases Tm.Detergent (SDS, Sarkosyl, Tween)Prevents nonspecific ionic interactions Insufficient detergent may result in background.of probe with the membrane.Excessive detergent may reduce sensitivity.Nonspecific nucleic acid Blocks nonspecific hybridization of nucleic Addition of nonspecific nucleic acid can decrease(herring or salmon sperm DNA)acid probe.nonspecific background by binding to non-specific regionson the membrane. Also, if the probe is a whole genomic probe,the herring sperm will block repetitive elements.Excessive amounts of nonspecific nucleic acid willreduce sensitivity.Formamide (deionized)Lowers the Tm of the nucleic acid hybridization.Formamide concentration up to 50% decreases the Tm ofthe nucleic acid hybridization and reduces the optimumhybridization temperatures.Protein solution (Blotto, Denhardt’s)Blocks nonspecific binding of probe to May reduce or increase background depending on thethe membrane.membrane used.Polymer accelerant Increases probe concentration by lowering May reduce or increase background depending on the (PEG, Dextran sulfate, PVP)the active water content.membrane used.Tm = melting temperatureTable 1: Hybridization Solution Components and EffectsThe stability of the nucleic acid duplexes and the stringency of hybridizationconditions determine the efficiency of hybridization. Several factorsdestabilize these hybrids by lowering their melting temperature. Thesefactors can be adjusted to favor formation of specific hybrids with minimalinterference from less specific hybrids. In any assay system, increasingstringency improves specificity with a corresponding loss in sensitivity;conditions should be optimized for specific applications. Table 1 describestypical hybridization solution components and their effects on hybridizationefficiency.The hybridization solution contained in Detector kits was formulated toinclude formamide. As a destabilizer formamide lowers the meltingtemperature of hybrids, increasing the stringency of the probe to targetbinding. Use of this agent results in minimal nonspecific hybridization;less optimization of washes is required by the end user. Unlike aqueoushybridization solutions, buffers containing formamide effectively minimizebackground to allow subsequent detection of single copy genes and lowexpressed transcripts. (Figure 5) Thus, these types of solutions can be appliedmore universally. Blotting procedures may also be expedited through the useof Formamide Hybridization Buffer, reducing an overnight hybridization to2 hours without impact on signal:noise ratio. Note that this is acceptablewhen detecting plasmid DNA or moderately expressed transcripts;however,overnight incubations are required for greatest sensitivity when detectinglow copy genomic DNA and rare mRNA.Detection MethodsDetection can be mediated either directly when using fluorescent haptensor indirectly with the use of binding proteins like antibodies or avidin/streptavidin as in the Detector system. The specific antibody or bindingprotein is coupled to an enzyme or fluorochrome and subsequently visualized Figure 5: Detection of a Low Expressed Transcript with Varying Hybridization Conditions.Duplicate lanes of 5 µg of total RNA from WEHI-231 untreated and anti-IgM treatedcells were electrophoresed on a 1% formaldehyde gel and transferred by a 2 houralkaline method to Biodyne B Nylon Membrane. The membrane was cut in half andhybridized with a biotinylated c-myc riboprobe in either formamide or aqueoushybridization buffer. Detection was carried out using the Detector AP ChemiluminescentBlotting Kit. 5A: When using a formamide-based solution, the c-myc gene was observedas a single band in the control sample (1) and the down-regulated treated sample (2).5B: Significant non-specific binding of the probe to the total RNA resulted on the blothybridized in an aqueous hybridization solution.5800-638-3167 • KPL, Inc. • 301-948-77557800-638-3167 • KPL, Inc. • 301-948-7755Detector Labeling KitsCatalog NumberKit SizeLabeling MethodSensitivityApplicationsRandom Primer DNA 60-01-0030 reactionsOnly 100 ng Biotinylation Kitpurified template needed per reaction.PCR DNA60-01-0130 reactionsAs little as 1 ng Biotinylation Kitgenomic template DNA can be amplified and labeled.RNA in vitro 60-01-0220 reactionsOne reactionTranscription generates enough Biotinylation Kitprobe to hybridize 48-96 blots.Detector Detection KitsCatalog NumberKit SizeDetection MethodSensitivityApplicationsAP Chemiluminescent 54-30-012000 cm 2AP-SA and CDP-StarBlotting Kit54-30-02500 cm 2Chemiluminescent SubstrateHRP Chemiluminescent 54-30-002000 cm 2HRP-SA and LumiGLOBlotting KitChemiluminescent SubstrateChromogenic In situ 60-03-0050 samplesHybridization KitTable 4: Choosing KPL Detector KitsExo- fragment of Klenow DNA polymerase extends primers by catalyzing the addition of nucleotidetriphosphate to the nascent probe from a mixture that includes biotin-dCTPSouthern, Northern and dot blotting; colony and plaque hybridization and in situ hybridization.Incorporation of biotin-dCTP via a thermostable DNApolymerase in the polymerase chain reaction.Southern, Northern, dot blotting; colony and plaque hybridization, and in situ hybridization.DNA located downstream of the RNA polymerasepromoter site is copied in a strand – specific manner into a RNA transcript in thepresence of ribonucleotides (biotin-UTP) and either T7 or SP6 RNA polymerase.HRP-SA and TrueBlue peroxi-dase substrate: Orcein and Eosin Y counterstains Southern and Northern blotting; mRNA in situ hybridization.Detection of single copy genes in 5 µg of genomic DNA,low expressed message in 1-5 µg total RNA or ß-actin in just 50 ng of total RNA after a 10-minute film exposure.Northern BlottingGenomic Southern blotting of single copy genes, plasmid DNA, dot blots, and PCR products.Detection of 0.3 pg DNA with a 15-minute film exposure.Southern blotting,bacterial colony and plaque hybridization dot blots.Sensitivityequivalent to FISH.DNA detection in cells, tissues, and metaphase chromosomes.Fluorescent In situ 60-05-0050 samples CY3/DAPI Hybridization KitCY3-SA: DAPI counterstain5-10 times greater fluorescence than FITC/TRITC labeled probes using the same filters.The remainder of The T echnical Guide to Non-Radioactive Labeling and Detection of Nucleic Acids consists of the detailed procedures for performing specific applications employing biotin and Detector kits. The following table (Table 4) summarizes the properties of the Detector product line, assisting in the selection of the appropriate system for your needs.DNA detection in cells,tissues, and metaphase chromosomes.Introduction to KPL’s Detector™Labeling KitsKPL offers three labeling approaches to the generation of biotinylated nucleic acid probes:• Detector Random Primer DNA Biotinylation Kit• Detector PCR DNA Biotinylation Kit• Detector RNA in vitro Transcription Biotinylation KitBoth random primer and PCR-mediated biotin labeling results in the net synthesis of DNA and amplification. Random primed labeling is catalyzed by Klenow polymerase, the large fragment of E. coli DNA polymerase. The Klenow polymerase lacks 5’ 3’ exonuclease activity of the holoenzyme but still contains the 5’ 3’ polymerase as well as the 3’ 5’ exonuclease proof-reading activity. During the polymerization reaction, Klenow polymerase incorporates not only the non-modified deoxynucleotides but also the hapten-modified substrates (e.g., biotin-dCTP), resulting in a DNA probe with high specific activity. PCR-mediated labeling of probes with biotin allows simultaneous amplification and labeling of DNA. Thermostable T aq DNA polymerase drives the PCR reaction, incorporating biotin into the PCR product via modified deoxynucleosite triphosphates. The end product is homogeneously labeled hybridization probes that can detect sub-picogram amounts of target sequences on blots.Probes generated by either random priming or PCR are typically used in Southern blots; they are also suitable for Northern blots. While DNA probes are commonly used in the detection of nucleic acids on membranes, RNA probes are an advantageous alternative and should be considered particularly when the visualization of low expressed genes is desired. In these cases, detection with riboprobes can be approximately 10 times more sensitive than DNA probes. This increase is accounted for by the great affinity of a riboprobe for the complementary sense strand of the mRNA being detected and the resulting higher stability of the RNA:RNA bond after hybridization.1 Additionally, single stranded RNA probes are not subject to the self-annealing that double-stranded DNA probes are, which decreases the availability of the DNA probes to bind to the immobilized target.Single-stranded RNA probes can be generated by in vitro transcription from RNA polymerase promoters such as SP6, T7 or T3. DNA located downstream of the RNA polymerase promoter site is copied in a strand-specific manner into an RNA transcript in the presence of ribonucleotides and the appropriate RNA polymerase. Because of the nature of transcription reactions, many copies of RNA are produced from the template DNA in a short time. Transcripts can be labeled during synthesis by incorporation of biotin during transcription. The incorporation of biotin-UTP by SP6 or T7 polymerase is very efficient, resulting in highly labeled RNA probes.The following protocols describe in detail the process of biotin labeling nucleic acid probes by random priming, PCR labeling and in vitro transcription using KPL’s Detector Biotin Labeling Kits. For additional assistance while using these systems, the labeling section of the Troubleshooting Chapter (Chapter 6) may be referenced.Detector Random Primer DNA Biotinylation Kit (Cat. No. 60-01-00)The Detector Random Primer DNA Biotinylation Kit provides a method for biotinylating DNA probes through incorporation of biotin-dCTP during random-primer extension.2, 3Six base random sequence oligonucleotides serve as primers for replication of the template DNA. The Exo- fragment of Klenow DNA polymerase extends the primers by catalyzing the addition of nucleotide triphosphate, from a mixture that includes biotin-dCTP, to the nascent probe. Large quantities of biotinylated DNA probes can be generated from a small quantity of template DNA. This labeling method results in the net synthesis of DNA. The use of Exo- Klenow polymerase allows for longer labeling reactions without the risk of degradation of the oligonucleotides.The components of this kit are optimized to maximize amplification of template DNA and sensitivity of target detection.4T emplate requirements – Optimal labeling occurs on templates that range from 300-1000 bp. The probe fragments generated vary in length from 100 to1,000 bases, with the average length being approximately 300 base pairs.Probe storage– Stable for at least one year when stored at -20°C. Because biotinylated probes stick to normal microcentrifuge tubes, it is recommended that probes be stored in low retention or siliconized tubes.Probe quantitation– A pre-labeled quantitation standard is included for determining the relative amount of DNA synthesized during the labeling reaction. The concentration of probe to be used in the hybridization buffer must be optimized for greatest sensitivity and minimal background.• Serial dilutions of quantitation standard and newly labeled probe arefixed to positively charged nylon membrane.• The standard and probe are detected using enzyme-labeled streptavidin, and chemiluminescent or chromogenic substrate.• Endpoint sensitivity of the samples is compared to the standard todetermine the amount of probe generated.Materials and EquipmentKit Components Product Code Volume2.5X Random Primer Solution 600-0001700 µL10X dNTP Mixture 600-0002175 µL Klenow DNA Polymerase 600-000335 µL (Exo-fragment)Control Template DNA 600-000425 µL Stop Buffer 600-0005250 µL DEPC Treated Water 50-86-03 1.0 mL Quantitation Standard 600-0007150 µLSufficient reagents are provided to perform 30 labeling reactions when followingthe protocol described below. Kit components are stable for a minimum of 1 year. Reagents must be stored at –20°C and kept on ice during use. Do not store kitsin a frost-free freezer.9800-638-3167 • KPL, Inc. • 301-948-7755Probe Labeling by Random Priming (continued)Steps Critical Points6. Incubate at 37°C for 1 – 4 hours. One hour is sufficient for most applications. For maximum yield of probe,allow the reaction to proceed for 4 hours. Generally, a reaction beginningwith 200-300 ng template DNA generates 5- to 10-fold amplification ofthe template in an hour, and 10- to 50-fold amplification after 4 hours.7. Add 5 µL Stop Buffer and mix.8. Proceed to Probe dilution, dot blotting It is imperative that the probe be quantitated. If too much probe is used inand detection for quantitation.hybridization, background could occur. If too little probe is used, sensitivitymay be reduced.9. Store at –20°C until ready to quantitate and use.See Probe Quantitation on page 18.NOTES ON…Probe Purification:• Following biotinylation, the newly labeled probes may be separated from unincorporated nucleotides by either ethanol precipitation or using KPL’s SpinPure filters (Catalog No. 60-00-53). This is not necessary for use of probes in Southern and Northern blot detection, as unincorporatednucleotides do not significantly increase background. However, if the probes are to be used for in situ hybridization, we do recommend the removal of unincorporated nucleotides. See page 17 for Probe Purification using the SpinPure filters.Detector™PCR DNA Biotinylation Kit Array The Detector™PCR DNA Biotinylation Kit provides a rapid method forbiotinylating DNA probes through incorporation of biotin-N4-dCTP via athermostable DNA polymerase in the polymerase chain reaction*.5-7Biotinylated probes generated using this kit are highly sensitive and allow forthe identification of low copy target sequences. The process of direct labelingduring PCR results in specific labeling and amplification of the sequence ofinterest even from crude DNA samples.The ratio of biotin dCTP to unlabeled dCTP is optimized to produce probeswith maximal biotin incorporation for detection of low copies or rare targetsin mRNA and plasmid or genomic DNA. Amplification and direct labelingfrom small samples of genomic (1 - 100 ng) or plasmid template (10 pg -1 ng) is most easily achieved by first optimizing the conditions for standardPCR before attempting to label the probe during the reaction.Direct detection of a PCR biotin-labeled fragment is also possible. Thebiotinylated PCR product is electrophoresed, transferred to membrane andsubsequently detected without a probe using enzyme-labeled streptavidin.A signal is then generated using the appropriate chromogenic orchemiluminescent substrate as outlined in any of the detection methodsof the Detector Kits. (See Figure 6)* Purchase of this kit does not constitute a license for PCR. A licensed polymerase andlicensed thermal cycler must be used in conjunction with this product. PCR is coveredby patents owned by Hoffman-La Roche, Inc. and F. Hoffman-La Roche, Inc.Detector PCR DNA BiotinylationAt A GlanceOptimize PCR Conditions With Unmodified dNTPsPrepare PCR Reaction MixPerform PCR ~25 cycles, 40 minutesQuantitate ProbeStore at -20°CStepsCritical Points1. Prepare the reaction mix in a sterile PCR tube inThaw the 10X Labeling Mix on the bench or warm it to room temperature in order as it appears below. Place the tube on ice while pipetting.your hand before use. Improperly thawed dNTPs may result in a failure to produce amplification product.Component Volume Final DEPC Treated Water variable add to 50 µl By adjusting the final concentration of the 10X Labeling Mix included in the 10X PCR Buffer 5 µl 1Xkit, probes of up to 1 kb in length may be generated. For probes less than 25 mM MgCl 24 µl 2.0 µM500 base pairs, a final concentration of 200 µM of each nucleotide is suggested 10X Labeling Mix 5 µl 200 µM each (5 µL/50 µL PCR reaction), and for probes greater than 500 base pairs, Primers1 µl0.5 µM each350 µM of each nucleotide is suggested (8.75 µL/50 µL PCR reaction).(if using the control primers)Taq DNA Polymerase variable 1.25 units/ 50 µl rxn Template1 µl1-10 ng(if using genomic DNA or the control 10 pg-1 ng template)plasmid DNATotal Mix50 µl2. Mix the tube by tapping gently and centrifuging briefly.Probe Labeling by PCRNOTES ON…Getting Started• The following protocol was designed specifically for the PCR labeling of the control template included with this kit. PCR reaction conditions should be optimized for each new template/primer set with unmodified nucleotides before use of this kit. Use the following protocol as a guideline only . • Caution should be taken to minimize introduction of contami-nating DNA and/or DNases to the PCR reaction that may result in amplification of non-specific product or no product. Always wear gloves, wash all work areas appropriately prior to beginning.• Allow all reagents to thaw out completely , then vortex briefly and spin down in a microcentrifuge before pipetting. Keep all reagents on ice while in use except for the 10X Labeling Mix. Pipette reagents slowly and carefully to avoid errors.• The buffers contained in and/or recommended for use with this kit are prepared according to the protocols listed at the end of this chapter, beginning page 20. Recipes for miscellaneous solutions can be found in the Appendix.Materials and EquipmentKit ComponentsProduct CodeVolume 10X Ribonucleotide Labeling Mix 600-001360 µL 10X Transcription Buffer 600-001060 µL ß-Actin Template 600-001716 µL RNase Inhibitor 600-001213 µL SP6 RNA Polymerase 600-001520 µL T7 RNA Polymerase 600-001420 µL DNase I600-001620 µL DEPC Treated Water 50-86-03 1.0 mL Quantitation Standard 600-0007150 µL Spin-Pure Filters60-00-535 filtersSufficient reagents are provided to perform 20 labeling reactions following theprotocol provided in this manual. All reagents must be stored at -20°C except for the Spin-Pure Filters that should be stored at room temperature. Do not store kits in a frost-free freezer. Kit components are stable for a minimum of 1 year from date of receipt when stored as instructed.Quantitation of the probe is essential as the concentration of probe used in the hybridization reaction is critical for greatest sensitivity and minimal background on a membrane.• Serial dilutions of quantitation standard and newly labeled probe are fixed to positively charged nylon membrane.• The standard and probe are detected using enzyme-labeled streptavidin, and chemiluminescent or chromogenic substrate.• Endpoint sensitivity of the samples is compared to the standard to determine the relative amount of probe generated.Probe storage – Stable for at least one year when stored at -20°C. Because biotinylated probes stick to normal microcentrifuge tubes, it is recommended that probes be stored in low retention or siliconized tubes.Detector RNA in vitro Transcription BiotinylationAt A GlancePreparation of DNA TemplatePrepare Labeling reaction mixIncubate at 37°C2 hoursAdd DNase IIncubate at 37°C 15 minutesQuantitate ProbeStore at –20°C[or -70°C for long-term storage]NOTES ON…Getting Started• All tubes and pipet tips should be autoclaved or purchased as RNase free prior to working with RNA.• All glassware and equipment used should be RNase free.• Always wear gloves when working with RNA because human skin contains abundant amounts of RNases.Detector RNA in vitro Transcription Biotinylation KitThe Detector ™RNA in vitro Transcription Biotinylation Kit provides a method for synthesizing biotin labeled RNA probes by in vitro transcription through incorporation of biotin UTP . Strand-specific probes may be generated using either the T7 or SP6 RNA polymerase.9The transcript reaction generates full length, single-stranded RNA probes that can be used in a variety of applica-tions, including membrane and in situ hybridization. The incorporation of biotin UTP by SP6 or T7 polymerase is very efficient, resulting in “hot”labeled RNA probes. These non-isotopic biotin labeled RNA probes are stable for at least one year. Single-stranded RNA probes hybridize more effectively to target molecules because they do not self-hybridize as DNA probes do.10RNA probes offer greater sensitivity than DNA probes because RNA-RNA or RNA-DNA hybrids are more stable than DNA-DNA duplexes in hybridization.Direct detection of a biotin-labeled RNA probe is also possible. The biotinylated transcript can be electrophoresed, transferred to a membrane, and subsequently detected without a probe using fluorochrome-labeled streptavidin or enzyme-labeled streptavidin and a chromogenic or chemiluminescent substrate as directed in any of the detection methods of the Detector kits.The components of this kit are optimized to maximize the synthesis of RNA as well as the sensitivity of target detection.T emplate requirements – In order to generate single stranded RNA probes, you must begin with a DNA template with SP6 or T7 promoter sequences upstream from the desired template sequence. T wo methods to prepare these types of templates are recommended and further detailed in the protocol:• Cloning the DNA into a vector with the SP6 and T7 promoter sequences on either side of the cloning site• PCR of the DNA template with the promoter sequences built into the primers.Kit control – A human ß-Actin DNA template is included in this kit to serve a two-fold purpose: 1) to act as a control for the integrity of the kit components, and 2) to generate a control probe for detection of ß-actin on human or mouse Northern blots. Synthesized in a strand-specific manner,an antisense transcript may be generated using T7 RNA polymerase and a sense transcript may be generated using SP6 RNA polymerase. The expected size of either transcript is ~400 bases. If the control probe is to be used in a Northern blot, the anti-sense (T7) probe must be used.Probe quantitation – The second kit control, the Biotinylated Quantitation Standard is used to quantitate the yield of biotin-labeled RNA probe.Probe Labeling by in vitro Transcription(continued)Steps Critical Points3. Mix the tube by flicking gently and centrifuge briefly.4. Place the tube at 37°C for 2 hours.5. Add 1 µL of DNase I, flick the tube gently, and centrifuge briefly.6. Incubate 37°C for 15 minutes.7. Place the tube on ice or store at –20°C until needed for Because of the high concentration of the probe and its susceptibility toquantitation and analysis.RNases, aliquoting of the probe for storage is recommended. For longterm storage, freeze at -70°C.NOTES ON… Probe Purification:• Following biotinylation, the newly labeled probes may be separated from unincorporated nucleotides by either ethanol precipitation or using KPL’s SpinPure filters (Catalog No. 60-00-53). This is not necessary for use of probes in Southern and Northern blot detection, as unincorporated nucleotides do not significantly increase background. However, if the probes are to be used for in situ hybridization, we do recommend the removal of unincorporated nucleotides. See below for Probe Purification using the SpinPure filters.Probe Purification (optional)Steps Critical Points1. Ensure that the sample reservoir is firmly placed into thefiltrate receiver.2. Add 1XTE to the probe to increase the volume. Do not tear the filter with a pipet tip.Pipette 50 - 500 µL of the sample into the sample reservoir.Cap the Spin-Pure filter and place into a microcentrifuge.3. Centrifuge at 5,000 x g for 15 minutes at room temperature.Continue centrifugation until filter is dry (a volume of 500 µLcan usually be concentrated in 20 minutes).4. If removing primers and nucleotides from amplified product,centrifuge at 14,000 x g.5. Recover sample from the filter with DEPC-treated wateror 1X TE by rinsing the surface. Highest yields result fromtwo rinses of 20 µL each.Gel Analysis of TranscriptSteps Critical Points1. Pre-heat the probe to 68°C for 5 minutes.The expected size of the transcript will be somewhat different whencomparing an RNA probe to a DNA marker lane, but an approximate sizeestimate can still be determined.2. Run 2-5 µL of the probe on either a 1X TBE agarose gel or a Biotinylated transcripts run larger than their unbiotinylated counterparts.formaldehyde gel containing 0.5 µg/mL ethidium bromide.Include DNA or RNA markers on the gel for proper size.3. Transfer the remaining aliquot of sample to a siliconized tube See Probe Quantitation on page 18.and store at 2–8ºC until quantitation.。
药物替代疗法英语作文
药物替代疗法英语作文Title: The Evolution of Pharmacological Alternatives in Therapy。
In the realm of medical science, the quest foreffective treatments has led to the exploration of diverse therapeutic avenues. Among these, pharmacological alternatives have emerged as a promising approach, offering new dimensions to conventional medical practices. Thisessay delves into the realm of pharmacological alternatives, exploring their significance, evolution, and impact on modern healthcare.Pharmacological alternatives, often referred to as drug substitutes or alternative medicine, encompass a broad spectrum of therapeutic interventions that diverge from conventional pharmaceuticals. These alternatives range from herbal remedies and dietary supplements to acupuncture and mindfulness-based practices. While their roots may lie in traditional healing systems, such as Traditional ChineseMedicine (TCM) or Ayurveda, pharmacological alternatives have gained traction in contemporary medical discourse dueto their perceived efficacy and reduced adverse effects.One of the primary drivers behind the growing interestin pharmacological alternatives is the increasing awareness of holistic health approaches. Unlike conventional medicine, which often focuses on symptomatic relief, pharmacological alternatives prioritize the restoration of balance within the body-mind system. For instance, practices like yoga and meditation not only alleviate physical ailments but also promote mental well-being, addressing the interconnectedness of physical and psychological health.Moreover, the rise of chronic diseases and thelimitations of conventional treatments have spurred the exploration of alternative therapeutic modalities. Conditions such as chronic pain, autoimmune disorders, and mental health disorders often pose challenges fortraditional pharmaceutical interventions, leading patients and healthcare providers to seek alternative solutions. In this context, pharmacological alternatives offer novelstrategies for managing chronic conditions, emphasizing preventive care and lifestyle modifications alongside symptomatic relief.The evolution of pharmacological alternatives has been shaped by a confluence of factors, including scientific research, cultural influences, and consumer demand. Over the years, rigorous scientific investigations have provided insights into the mechanisms of action and therapeutic benefits of various alternative treatments. For example, studies have elucidated the anti-inflammatory properties of certain herbs, the neurobiological effects of acupuncture, and the stress-reducing effects of mindfulness practices.Furthermore, cultural traditions and indigenous knowledge have played a significant role in preserving and disseminating pharmacological alternatives. Practices such as traditional herbal medicine have been passed down through generations, enriching the diversity of therapeutic options available to patients worldwide. In recent decades, initiatives aimed at integrating traditional healing practices into mainstream healthcare systems have gainedmomentum, fostering cross-cultural exchange andcollaboration in the field of pharmacological alternatives.The impact of pharmacological alternatives on modern healthcare extends beyond individual health outcomes to encompass broader societal and economic dimensions. By promoting preventive care and self-care practices, pharmacological alternatives have the potential to reduce healthcare costs and alleviate the burden on overburdened healthcare systems. Moreover, these alternatives empower individuals to take an active role in their health andwell-being, fostering a sense of autonomy and empowerment.However, it is essential to approach pharmacological alternatives with a critical and evidence-based perspective. While many alternative therapies show promise in clinical settings, not all interventions are supported by robust scientific evidence. Moreover, the lack of standardized regulation and quality control poses challenges in ensuring the safety and efficacy of alternative treatments. Therefore, interdisciplinary collaboration between conventional medicine and alternative healthcare providersis crucial in navigating the complexities of pharmacological alternatives and integrating them into comprehensive patient care plans.In conclusion, pharmacological alternatives represent a dynamic and evolving frontier in modern healthcare,offering innovative approaches to health and wellness. From ancient healing traditions to contemporary scientific discoveries, the evolution of pharmacological alternatives reflects the diverse tapestry of human knowledge and experience. By embracing a holistic perspective and fostering collaboration across disciplines, we can harness the potential of pharmacological alternatives to promote health, alleviate suffering, and enhance the quality oflife for individuals worldwide.。
胶原蛋白流失的速度英语作文
胶原蛋白流失的速度英语作文Collagen loss is a natural process that occurs in the body over time, but the speed at which it happens can vary.One factor that influences the speed of collagen loss is age. As we get older, the body's natural ability to produce collagen decreases.Excessive sun exposure can accelerate the loss of collagen. Prolonged exposure to UV rays can damage the skin and affect collagen production.Unhealthy lifestyle choices, such as a poor diet, smoking, and lack of sleep, can also contribute to faster collagen loss.Stress can have an impact on the body's collagen production and increase the rate of loss.The environment we are in can also play a role. Pollution and other external factors can damage the skin and lead to quicker collagen loss.Genetics can determine how quickly collagen is lost. Some individuals may be more prone to faster collagen loss due to their genetic makeup.The rate of collagen loss can also be influenced by certain medical conditions or medications.To slow down the speed of collagen loss, it is important to take proper care of the skin. This includes using sunscreen, maintaining a healthy diet, reducing stress, and getting enough sleep.In conclusion, understanding the factors that affect the speed of collagen loss can help us take measures to protect our skin and maintain its elasticity and firmness.。
Illumina cBot自动克隆扩增系统说明书
The Best Next-Gen Sequencing Workflow Just Got BettercBot is a revolutionary automated clonal amplification system at the core of the Illumina sequencing workflow (Figure 1, upper panel). cBot replaces a lab full of equipment with a single compact device, deliver-ing unsurpassed efficiency and ease of use for the highest quality sequencing results.With cBot, hands-on time is reduced to less than 10 minutes, com-pared to more than six hours of hands-on effort for emulsion PCR methods. The process of creating sequencing templates is complete in about four hours, compared to more than 24 hours for emulsion PCR-based protocols (Figure 1, lower panel).Breakthrough System for Cluster GenerationThe Illumina sequencing workflow is based on three simple steps: libraries are prepared from virtually any nucleic acid sample, amplified to produce clonal clusters, and sequenced using massively parallel synthesis. The cBot clonal amplification system has innovative features that eliminate user intervention, reduce potential failure points, and increase sequencing productivity.TruSeq Cluster Generation reagents are packaged in ready-to-use96-well plates, completely removing reagent preparation errors, potential sources of contamination, and decreasing storage require-ments. cBot features a single unique, plate-piercing manifold for intervention-free runs. Cluster generation occurs within the sealed, eight-channel Illumina flow cell, bypassing the frequent handling and contamination issues inherent to emulsion PCR-based protocols. cBot is capable of processing > 96 samples within a single flow cell, resulting in substantial cost savings without incremental effort and wasted reagents. Innovative instrument features ensure seamless operation for your sequencing workflow (Figure 2).Better Results with Less EffortcBot software enhancements and user interaction features ensure high productivity:• Integrated 8-inch touch screen provides simplified operation in a small, lab-friendly footprint• On-screen, step-by-step instructions with embedded multimedia help enable user operation with no prior training • Real-time progress indicators provide at-a-glance monitoring • Remote monitoring allows a single user to manage multiple systems from any web browser or phone• Status emails are sent when the run is complete or when intervention is requiredcBot Cluster Generation ProcessPrior to sequencing, single-molecule DNA templates are bridge amplified to form clonal clusters inside the flow cell. (Figure 3).cBotFully automated clonal cluster generation for Illumina sequencing.Illumina cBot Highlights• Fast, Efficient Workflow:Amplify > 96 samples in ~4–5 hours with < 10 minutes ofhands-on time• Easiest to Use:Pre-packaged 96-well TruSeq™ reagents, and simple touch screen interface simplifies operation• Innovative System Design:Real-time fluidic monitoring, integrated system sensors and remote monitoring ensure robust instrument operation• Highest Quality Results:Improved chemistry generates higher density clusters and sequencing accuracy LibraryPreparation SequencingCluster GenerationEight-channel flow cell reduces risk of contamination and eliminates the needfor extra equipment Manifold clamps for leak-free connections and superior thermal contactTouch screen monitor simplifies operation and provides real-timeImmobilization of Single-Molecule DNA TemplatesHundreds of millions of templates are hybridized to a lawn of oligo-nucleotides immobilized on the flow cell surface. The templates are copied from the hybridized primers by 3’ extension using a high-fidelity DNA polymerase to prevent misincorporation errors. The original templates are denatured, leaving the copies immobilized on the flow cell surface.Isothermal Bridge AmplificationImmobilized DNA template copies are amplified by isothermal bridge amplification. The templates loop over to hybridize to adjacent lawn oligonucleotides. DNA polymerase copies the templates from the hybridized oligonucleotides, forming dsDNA bridges, which are dena-tured to form two ssDNA strands. These two strands loop over and hybridize to adjacent oligonucleotides and are extended again to form two new dsDNA loops. The process is repeated on each template by cycles of isothermal denaturation and amplification to create millions of individual, dense clonal clusters containing ~2,000 molecules. Linearization, Blocking, and Primer HybridizationEach cluster of dsDNA bridges is denatured, and the reverse strand is removed by specific base cleavage, leaving the forward DNA strand. The 3’-ends of the DNA strands and flow cell-bound oligonucleotides are blocked to prevent interference with the sequencing reaction. The sequencing primer is hybridized to the complementary sequence on the Illumina adapter on unbound ends of the templates in the clusters. The flow cell now contains >200 million clusters with ~1,000 mol-ecules/cluster, and is ready for sequencing.SummaryIllumina sequencing with cBot automated cluster generation sets the new standard for simplified next- generation sequencing. Ready-to-use reagents, smart instrumentation improvements, and new cluster generation chemistry offers significant advantages over emulsion PCR-based workflows and promotes even higher data density and sequencing accuracy. By streamlining the critical clonal amplification step in the next-generation sequencing workflow, Illumina continues to accelerate your landmark discoveries and publications.Ordering InformationDescriptioncBotCatalog No.HiSeq System Genome AnalyzercBot Instrument Includes cBot, flow cell adapter plate,one year warranty, user manualSY-301-2002cBot Flow Cell Manifold (Optional)SY-301-2014TruSeq Single-Read Cluster Generation Kits include flow cell,reagent plate, manifold, user instructionsGD-401-3001GD-300-2001TruSeq Paired-End Cluster Generation Kits include flow cell,reagent plate, manifold, PE reagents, user instructionsPE-401-3001PE-300-2001Illumina, Inc. •9885TowneCentreDrive,SanDiego,CA92121USA•1.800.809.4566toll-free•1.858.202.4566tel•************************• For research use only© 2011 Illumina, Inc. All rights reserved.Illumina, illuminaDx, BeadArray, BeadXpress, cBot, CSPro, DASL, Eco, Genetic Energy, GAIIx, Genome Analyzer, GenomeStudio, GoldenGate, HiScan, HiSeq, Infinium, iSelect, MiSeq, Nextera, Sentrix, Solexa, TruSeq, VeraCode, the pumpkin orange color, and the Genetic Energy streaming bases design are trademarks or registered trademarks of Illumina, Inc. All other brands and names contained herein are the property of their respective owners. Pub. No. 770-2009-032 Current as of 27 April 2011at the address below.Laser radiationDo not stare into the visible-light beam of the barcode scanner. The barcode scanner is a Class 2 laser product.SY-301-2002Instrument ConfigurationCE Marked and ETL Listed instrument, Installation, setup, and accessoriesInstrument Control ComputerMini-ITX Board with Celeron M Processor 1 GB RAM, 80 GB Hard Drive Windows Embedded OSIntegrated 8” Touch Screen Monitor Operating Environment Temperature: 22°C ± 3°CHumidity: Non-Condensing 20%–80%Altitude: Less than 2,000 m (6,500 ft)Air Quality: Pollution Degree Rating of II For Indoor Use Only LaserClass 2 Laser: 630 –650 nm DimensionsW×D×H: 38 cm × 62 cm × 40 cm Weight: 34 kg Crated Weight: 36 kg Power Requirements100−240V AC 50/60 Hz, 4A, 400 Watts。
Analysis of compressive fracture of three different concretes by means
Analysis of compressive fracture of three different concretes by means of 3D-digital image correlation and vacuum impregnationDaniel Caduff,Jan G.M.Van Mier *Institute for Building Materials ETH Zurich,8093Zurich,Switzerlanda r t i c l e i n f o Article history:Received 12August 2009Received in revised form 6January 2010Accepted 20January 2010Available online 25January 2010Keywords:Concrete FractureUniaxial compression3D-digital image correlation Vacuum impregnation Crack orientationa b s t r a c tFracture of three different concretes under uniaxial compression was investigated.The three different materials were normal concrete,high strength concrete and foamed cement,with uniaxial strength vary-ing between 8and 92MPa.The prismatic specimens were loaded between two different types of loading platens,providing maximum boundary restraint (rigid steel platens)or almost zero boundary restraint (Teflon (PTFE)sandwich inserts).Fracture propagation was measured using 3D-digital image correlation where emphasis was placed on measuring crack development just before and at peak stress.The stress at first cracking is dependent on the resolution of the digital image correlation technique,and as such not very well defined.The frictional restraint of the loading platens is recognized in the failure modes,in par-ticular in the direction and structure of the main cracks.Also,the results for normal and high strength concrete underscore previous results:high friction results in a higher uniaxial compressive strength.This finding was not confirmed for foamed cement,where both loading systems gave approximately the same compressive strength.Likely the Teflon inserts do not function properly on the highly porous surfaces of the foamed cement specimens.The direction of the main cracks at peak is defined before the softening regime is entered:cracks have a vertical orientation when loading is applied through Teflon inserts,whereas ‘en-echelon’tensile cracks forming inclined shear bands are observed for rigid loading platens.The results from 3D-digital image correlation were confirmed by results from vacuum impregnation with fluorescent epoxy after test termination.Although digital image correlation is capable of showing cracks at relatively early stages of loading,the vacuum impregnation technique shows all the fine detail and crack branches,but this can,unfortunately,only be done once for each specimen.Therefore,combining the two different techniques is more appropriate.Ó2010Elsevier Ltd.All rights reserved.1.IntroductionCompressive fracture of concrete remains an elusive phenome-non.One of the main problems in studying fracture is that when crack size approaches that of the specimen (or structure),bound-ary effects,influences of specimen size and the fracture properties become closely interdependent and are difficult if not impossible to separate.This problem has not been resolved to date,in partic-ular not for fracture of cohesive frictional materials like concrete,where fracture is not characterized by the nucleation and growth of a single macroscopic crack,but where in some stages of the frac-ture process a multitude of small short microcracks appear.The fracture process in compression can be divided into four different stages [1,2]as follows:(O)linear elastic stage,(A)microcracking,(B)macrocrack growth and (C)bridging and frictional restraint.Stages (O)and (A)are associated with the rising branch of thestress–strain curve leading to maximum stress;stages (B)and (C)are related to the degradation of the specimen in the post-peak softening regime.In stage (O)it is assumed that no damage occurs and the material behaves linear-elastic.This is an assumption be-cause due to the manufacturing process of concrete early damage is almost unavoidable,and a truly linear regime will not exist.When external load increases local (tensile)stress concentrations cause the development of small microcracks that can still be ar-rested by stiffer and strong elements in the heterogeneous material structure.However,upon increasing external load,at some mo-ment the material structure cannot arrest further crack propaga-tion and localization of damage in a single (few)large crack(s)occur(s).We enter stage (B)of the aforementioned fracture pro-cess.The carrying capacity of the specimen (or rather structure)is decaying rapidly,and some residual carrying capacity remains due to bridging and frictional interlock in the macrocracks (stage (C)).Macrocrack propagation (stage (B))is affected by boundary conditions and specimen size and shape,and must thus be seen as a structural property.Bridging (stage (C))is affected by material composition.For example the size of the largest aggregates as well0958-9465/$-see front matter Ó2010Elsevier Ltd.All rights reserved.doi:10.1016/j.cemconcomp.2010.01.003*Corresponding author.E-mail addresses:caduffd@ethz.ch (D.Caduff),jvanmier@ethz.ch ,jvanmier@x-s4all.nl (J.G.M.Van Mier).Cement &Concrete Composites 32(2010)281–290Contents lists available at ScienceDirectCement &Concrete Compositesjournal homepage:w w w.e l s e v i e r.c o m /l ocate/cemconcompas the strength and stiffness contrast of the aggregate and matrix define the width of the localized shear-band in compression,and thus the residual stress in the softening regime.All these matters have been presented in detail in the aforementioned papers,and will not be further debated here.The intention of the present paper is to further investigate the damage patterns in different types of concrete subjected to uniaxial compression.Of particular interest is the transition from the microcrack stage(A)to the macrocrack stage(B),which happens around peak stress.Identifying the condi-tions under which the localized macrocrack(s)nucleate and grow is of great importance for engineering new concretes with im-proved fracture behaviour and for predicting the failure strength of concrete structures.Modeling the fracture process is possible using micromechan-ics,starting from the particle level of concrete(see for example [3–12]).From these microscopic models,all the four aforemen-tioned stages in the fracture process follow directly.The models are usually based on a limited number of well understood and physically-based parameters.Although these models are computa-tionally expensive,they do not suffer from the problem encoun-tered in cohesive fracture models in the spirit of the Fictitious Crack Model[13],and the extension to compressive fracture, [14].In such models the post-peak regime,i.e.the softening curve is assumed to be a material property in-spite of the un-deniable strong boundary and size effects.In principle testing infinitely large specimens would be required to extract the parameters needed in cohesive crack models,but this runs rapidly into practi-cal problems.Testing2-(m)large plain-concrete specimens is al-ready a major effort in the best-equipped laboratories in the world[15].In the present investigation three different concretes of varying quality(defined according their uniaxial compressive strength) and two different boundary conditions were tested.Prismatic spec-imens were subjected to uniaxial compression between either rigid steel loading platen(the usual standard loading condition in com-pression tests)or between Teflon(PTFE)friction-reducing pads(as defined by RILEM TC148[16]).Thus,the two boundary conditions are‘high frictional restraint’and‘low frictional restraint’,which is known to have a significant effect on the failure mode and fracture strength in compression[17].The three concretes tested were a normal-strength concrete(35–40MPa),a low-strength foamed ce-ment(8–10MPa)and a high strengthfibre-reinforced concrete (90–95MPa),which span a large range in behaviour from brittle to quasi-brittle to quasi-ductile.The development of cracks was followed in real time using a3D-digital image correlation tool (VIC-3D),which allowed monitoring simultaneously two side sur-faces of a prism under load.At the end of each test the specimens were subjected to vacuum impregnation,a well-proven method that allows for quite detailed crack detection in the samples.Since this is a destructive method,it can only be used for comparison with the VIC-3D analysis at the end of each experiment.The paper has been organized as follows.After a brief summary of the used materials and specimen manufacturing methods,the digital image correlation method is explained in some detail.Next the results are presented,in particular the fracture patterns from the VIC-3D measurements in relation to the measured load–dis-placement curves.The crack patterns from VIC-3D are compared to those from vacuum impregnation in the last load-step.Finally the results are discussed and compared to results from others and conclusions are drawn.2.Materials and specimensUniaxial compression tests were carried out on prisms with a height/diameter ratio of2.The specimens were cast horizontally in steel moulds of size280Â70Â70mm3.After24h the spec-imens were de-moulded and for the next34days cured in a cli-mate room(T=20°C and RH=95%).The specimens were cut and ground after7days to the required slenderness of2.Three sides of each specimen were ground,namely the two surfaces that were viewed with the VIC-3D system,and the casting sur-face.In order to avoid problems with the VIC-3D system larger pores at the viewed surface werefilled with a liquid cement suspension.The effect offilling these larger pores can be seen from Fig.1.Afterfilling the pores,the viewed surfaces were coated with a white paint and sprayed with black aerosol to produce the required surface condition for the digital image correlation.Three different concretes were tested.Characterized by their compressive strength they covered the entire spectrum from high to very low strength.The expected fracture behaviour was from brittle to quasi-brittle to quasi-ductile.Table1shows the three concrete compositions.The foamed cement was produced follow-ing the procedure outlined in Meyer et al.[18].The high strength concrete contained some PVAfibres(Kuraray;12mm long, 0.2mm diameter)and air-entrapping agent to prevent extreme brittle(explosive)failure behaviour and possible damage to the cameras of the VIC-3D system.The maximum aggregate size in the high strength concrete was4-mm.Normal-strength concrete contained16-mm river gravel.The ratio D/d max was therefore 70/16,which meets minimum demands for having specimens with a representative volume.Some fresh and hardened concrete properties are listed in Table2.The compression strength is the average of three tests on prisms(H/D=2)between rigid steel loading platens.3.Test method and measurement methods3.1.Loading conditionsThe uniaxial compression tests were performed in a Wal-ter+Bai servo-hydraulic testing machine with a capacity of 4000kN.The loading was applied in force control and the loading rate varied between the three different concretes to ensure that the time to peak-load was the same for each mixture.A load-cell with a measurement range up to1000kN was used.For each con-crete,three specimens were loaded between rigid steel platens and three specimens were loaded using the friction-reducing Tef-lon(PTFE)sandwich inserts proposed by RILEM TC148SSC[16]. For the rigid steel loading platen a10mm thick steel platen with exactly the dimensions of the concrete prisms was inserted be-tween the specimen and the machine loading platens.The Teflon sandwich consists of2Teflon sheet of0.10mm thickness with a 0.05mm thick greasefilm in between.The total sandwich was thus0.25mm.Fig.2shows the applied loading history for each specimen.It was attempted to increase the force in every cycle without reach-ing the maximum load the specimen could sustain,except in the last cycle.The last cycle was stopped as soon as a decline in load of5kN was measured.At that moment it was assumed that the peak of the stress–strain curve was reached,and that upon further loading the softening regime would be reached.The whole surface of two sides of each prism was recorded with the two cameras of the digital image correlation system during the 1st and4th cycle.The image size was150Â150mm,thus covering the complete specimen length of140mm.During the2nd and3rd cycles,the upper and lower parts of the two surfaces were re-corded separately with an image size of90Â90mm in an attempt to improve the resolution in(pixel/mm).282 D.Caduff,J.G.M.Van Mier/Cement&Concrete Composites32(2010)281–2903.2.3D-digital image correlationThe digital image correlation system used was the VIC-3D sys-tem provided by LIMESS.3D-digital image correlation is based on both digital image correlation [19,20]and stereovision,and was developed at the end of last century (see [21–24]).The correlation scores are computed by measuring the similarity of a fixed sub-set window in the first image (reference image on the unloaded spec-imen)to the shifting sub-set window in the second image (of the loaded specimen).A summary of the algorithm can be found in [25].The main components of the system used are the two CCD cameras with a resolution of 2048Â2048pixels.The cameras are fixed on a traverse in vertical position as indicated in Fig.3.The cameras can be equipped with lenses of varying focal length,and it is possible using the same system to record the dis-placements of surfaces of a few millimeters to severalmeters.Fig.1.Same concrete surface without (left)and with (right)filled air voids.Table 1Concrete mixture ponent aNormal concrete High strength concrete Foamed cement Cement type CEM I 42.5N CEM I 42.5N CEM I 52.5R Cement 344995716Water1552632720–4mm sand953704–4–8mm aggregate 395––8–16mm aggregate 558––Fly-ash–157–Micro-silica–44–Plasticizer (Glenium ACE 30)–26–Air-entraining agent –10–PVA fibres –26–Protein foam––40aAll quantities in kg/m 3.Table 2Fresh and hardened concrete ponentNormal concrete High strength concrete Foamed cement Slump flow (cm)4296–Small slump flow (cm)––15.5Density (kg/m 3)236222781013f c (MPa)92.137.08.5Fig.2.Loading history for the tested specimens.D.Caduff,J.G.M.Van Mier /Cement &Concrete Composites 32(2010)281–290283The actual dimensions of the images depend on the used lens and the distance between the lens and the recorded surface.The angle between the two cameras must be greater than 30°;otherwise the out-of-plane displacement is calculated with a rather poor accuracy.Calibration of the system is essential in or-der to determine the best possible position of the two cameras,whereas the quality of the calibration also determines the accu-racy of the digital image correlation.As mentioned,for calculating the displacements with digital image correlation a reference image (at zero-load)and an image after deformation must be recorded.Before the software VIC-3D calculates the displacements between these two images an Area Of Interest (AOI)has to be set on the reference image (see Fig.4).Three parameters affect the displacement and strain calculation,namely the sub-set size,the grid step-size and the strain windowsize (also called the neighboring matched points).The effects of varying these parameters on the displacement and strain calcula-tion are explained in Robert et al.[25].It should be mentioned,that the image noise cannot be elimi-nated completely.The magnitude in (mm)for displacement (or in (%)for strain)of the image noise depends on the resolution in (pixel/mm)for the same parameter setting.Therefore,the image noise has a constant value in (pixel)for constant parameter set-tings while varying the resolution.In summary of Robert et al.[25],the measuring accuracy im-proves with increasing values of the three parameters.The image noise decreases when these parameters increase.When uniform displacements are measured,the parameter sizes have no influ-ence on the mean values of the displacements and the strains over the AOI.However,the situation changes when a material like con-crete is considered.The displacement fields become very irregular,especially during crack propagation.In order to detect the irregu-larities,the parameters should be set as small as possible.Small strain concentrations might indicate where cracks nucleate and grow.The obvious problem is the increasing image noise,which may be circumvented by decreasing the size of the image section.This of course has the disadvantage that not the fracturing of the entire specimen can be observed in a single image.If the ratio in (pixel/mm)is increasing when the image section is scaled down,and the image noise is independent of the magnification,then the image noise in (mm)becomes smaller (assuming that the val-ues of the three parameters are identical during the calibration of the displacements and strains in both cases).To verify the above an image section of size 240Â240mm from a surface of a concrete prism was recorded during a compression test.The test was terminated after reaching a load of 600kN,which is about 93%of the maximum stress for this particular specimen.After un-loading,the specimen was loaded again till 600kN but the image section was reduced to 120Â120mm.In Fig.5the prin-cipal strains are compared.The background color relates to the va-lue of the image noise,which has the same value in (pixel)forbothFig.3.Setup of the VIC-3D camera in front of the loading rig.The orientation of the specimen allowed viewing two surfaces of a prism simultaneously.The two cameras were placed vertically,asshown.Fig. 4.Location of two Area’s Of Interest (AOI)on the recorded image of the specimen.284 D.Caduff,J.G.M.Van Mier /Cement &Concrete Composites 32(2010)281–290image sizes.The correlation parameters were the same for both calculations.In the large image the shapes of the cracks are not very well reproduced (i.e.are hard to distinguish from the image noise)and the crack widths are over-estimated.With reduced im-age-section-size cracks are more clearly visible,at smaller crack widths and the crack shape seems better preserved.The dimension of the image section has thus an influence on the stress-level where first cracking is detected and cracks can be separated from the image noise.A number of other factors have influence on the accuracy of the displacement measurement.When the light conditions are chan-ged during recording,the correlation function calculates the wrong displacements because the shifting sub-set windows cannot be de-tected correctly.Vibrations and movements of the system during recordings also results in erroneous displacement values.The cor-relation function malfunctions in regions where the surface has rough edges.Therefore in the present experiments the AOI was kept away from the specimen edges,i.e.was not located at the intersection of the specimen and the background (see Fig.4).There are also some advantages of working with a full 3D sys-tem.In comparison to working with two-dimensional correlation software,with 3D DIC the surfaces of the specimen do not have to be planar,and the cameras need not to be positioned perpendic-ular to the observed surface(s).More than one surface of a speci-men can be recorded at the same time.With a 3D-system every movement in 3D-space can be measured and the out-of-plane deformation of a surface does not lead to errors in the calculation.4.ResultsIn this section the results from the experiments are summa-rized.After a brief overview of the stress–strain curves of the three different concretes and the determination of the stress-level at first cracking,difference in fracture behaviour between the three differ-ent concretes as seen with the 3D-digital image correlation are presented.The specimens were subjected to vacuum impregnation after the last loading cycle,and the crack patterns from the impreg-nated samples are compared to the digital image correlation results.4.1.Stress–strain curvesIn the first and fourth loading cycle (see Fig.2),the stress–strain curves of the tested concretes could be determined because the surface strains from two sides of a prism were measured withthe digital image correlation system with an image size of 150Â150mm.To calculate the axial and lateral strain,a release window was chosen in the middle of the prism with a height of 70mm.The average axial and lateral strains were calculated over the area of the release window.A typical stress–strain curve for foamed cement is shown in Fig.6.The ascending branch of the ax-ial strain is linear up to a level of about 30%of peak stress.Initially the lateral strains are of the same order of magnitude as the image noise,and the lateral strain curve is not quite smooth.This effect is reinforced when the volumetric strain is plotted (e vol =e axial +2Áe lat-eral ).Obviously the Poisson’s ratio,the Young’s modulus and the specimen’s volume change can be derived directly from the digital correlation data.In Fig.7the stress–strain curves from the first loading cycle are shown for all concretes tested.All test results lie between the two curves shown for each concrete type.The initial modulus for nor-mal concrete and for high strength concrete lies within the same range.The modulus for foamed cement is clearly much lower (see also Fig.8a).The strength differences are obvious,where it should be recalled that in the first cycle specimens were not loaded to peak stress.In the experiments it was found that thedifferencesparison of the crack pattern on the surface of a concrete prism loaded to 600kN using an image size of 240Â240mm (left)and 120Â120mm (right).Note that the higher resolution image was made after un-loading and re-loading the specimen for a second time to the original loading of 600kN.Fig.6.Stress–strain curve,measured during the 4th cycle.D.Caduff,J.G.M.Van Mier /Cement &Concrete Composites 32(2010)281–290285in this initial part of the ascending part of the stress–strain curves was not different when either rigid steel loading platens or Teflon friction-reducing pads were used.In Fig.8the measured Young’s modulus in the first loading cycle and the maximum compressive strength for the three concretes,loaded between rigid steel platens and Teflon inserts are gathered.The Young’s modulus of the high strength concrete is lower than that for the normal concrete because it contains no rough aggre-gates.Foamed cement has a much lower Young’s modulus than the two other concretes,which is caused by the large air content and the aggregate content of the normal concrete is higher.Differ-ences between loading system on Young’s modulus are small and negligible.Note that the scatter band for the Teflon inserts is rela-tively large,especially for the normal concrete and the high strength concrete.The uniaxial compressive strength is clearly more affected by the boundary restraint exerted by the loading platens,and confirms experiments by others.Surprising is that for foamed cement the differences between rigid steel platens and Teflon inserts decreases substantially,and it can be concluded that the loading platen friction has no influence on the strength of foamed cement.4.2.Stress at first crackingIn Table 3are listed the mean value and the standard devia-tion of the stress (as percentage of the peak stress)at first crack observation for the four load cycles.No distinction is made be-tween concretes,or between loading condition (low and high fric-tional restraint)because the observed differences were very small.The stress-levels are normalized to the peak stress for every specimen to allow for comparison of results.Beginning at the 1st load-cycle,the first crack is observed at an average stress of 71%with a standard deviation of 8%.The stress-level drops to 44%in the second load-cycle.This is not only caused by increas-ing damage in the specimen,which would decrease the stress at which crack propagation continues,but also because the recorded image section is reduced (see Section 3.1).The reduction of the image size increases the resolution in (pixel/mm)and also the measurement accuracy.Between the 2nd and 3rd cycle the re-sults can be compared directly since the image size has not been changed.Between the 3rd and 4th cycle the stress-level at first cracking increases again since the image size was brought back to the original size at the first paring the results of the 1st and 4th cycle shows that the first crack level decreased from 71%to 44%.This is directly the result from the three addi-tional load-cycles.These results are in agreement with earlier observations of concrete loaded to cyclic compression;see for example in Van Mier [26].4.3.Digital image correlationThe following images were recorded at the peak of the 4th cycle for each specimen.The settings of the parameters in the VIC-3D system were the same for every analysis,with a sub-set of 21,a step size of 5and a strain window size of 5.The largestmaximumFig.7.Enveloping curves of the axial strain for allmixtures.Fig.8.(a)Young’s modulus for the three mixtures:results from tests between rigid steel platens and Teflon platens are shown separately.(b)Effect of loading platen on the uniaxial compression strength for three different concretes tested.Table 3Comparison of stress-level at first crack.Stress level as percentage of peak stress (%)Mean valueStandard deviation 1st cycle 70.568.022nd cycle 44.449.843rd cycle 36.8814.014th cycle43.5316.56286 D.Caduff,J.G.M.Van Mier /Cement &Concrete Composites 32(2010)281–290principal strain is indicated by white.In interpreting these results one has to be careful since the maximum principal strains do not have the same value in all images.The image noise lies between 0.10%and0.15%for the used parameter settings.It is assumed that every visible strain concentration indicates a crack.Fig.9a and b shows the cracking in two normal concrete prisms loaded between high friction boundary conditions and low friction boundaries respectively.Steel platens were used in Fig.9a.Two global‘shear-bands’can be identified on the two recorded surfaces of the prism.The‘shear bands’(inclined cracks)extend over the entire height of the specimen.The‘shear bands’appear to consist of vertical splitting cracks.The cracks with the largest principal strains have a vertical orientation;cracks with a more inclined ori-entation in general show smaller principal strains.Cracks seem to propagate from the specimen’s corner only;along the edges hardly any further cracks were found,which may be the result of the high friction end condition.In Fig.9b the crack patterns obtained from a test on normal concrete loaded between Teflon inserts is shown (low friction).In the normal concrete specimens large air voids were notfilled with the liquid cement suspension,and the effect is directly visible.High principal strains are calculated at the loca-tion of the air voids,but luckily due to their round shapes the air voids can easily be distinguished from the cracks.In this test ver-tical splitting cracks were detected on the left surface.The cracks did not appear to‘line-up’in a‘shear band’as‘en-echelon’tensile cracks.Moreover cracks not only propagate from the corners but also along the edge(see the right surface,along the bottom).This may be a direct consequence of the low friction boundary condition.The high strength concrete prism,loaded between stiff steel platens,is shown in Fig.10a.The behaviour is quite comparable to that of the normal concrete in Fig.9a.In particular on the right surface the shear band seems to manifest itself in the same way as in the normal concrete prism.The shear band does not extend over the full height of the specimen,but is composed of a number of vertical splitting cracks.The black re-gion on the top side of the right surface,near the main crack is the result of spalling.The VIC-3D software automatically ignores this region.The high strength concrete specimen loaded between Teflon shows a number of vertical splitting cracks, mostly along the top side of the specimen(Fig.10b).The lon-gest cracks propagate almost over the length of thespecimen;Fig.9.Crack patterns in a normal concrete at peak-load of cycle4:(a,c)loaded between rigid steel platens and(b,d)loaded between Teflon.Figures a and b show the strain concentrations visualised with digital image correlation;figure c and d shows crack patterns after vacuum impregnation.D.Caduff,J.G.M.Van Mier/Cement&Concrete Composites32(2010)281–290287。
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Improved Glottal Closure Instant Detector Based OnLinear Prediction And Standard Pitch ConceptCheol-Woo Jo 1, Ho-Gyun Bang 1, William A.Ainsworth 21 Dept. of Control and Instrumentation Engineering, Changwon National University, Koreacwjo@sarim.changwon.ac.kr2 Dept. of Communication and Neuroscience, Keele University, U.K.coa01@ABSTRACTThis paper proposes an improved method of glottal closure instant detection using linear prediction and standard pitch concept. The main improvements are on its speed of computation and error reduction on position finding for the cases that were not possible or caused many errors using previous methods.Our method can resolve the problems occurring in current methods to some extent. The false location detection rate is reduced to its inherent interpolation capability. Also the amount of computation is reduced. Another benefit from our method is that it does not need additional post processing to find peaks or smoothing of the pitch tracks. All is contained in itself. Also we compared results among three different kinds of linear prediction based pitch detectors.1. INTRODUCTIONDetecting glottal closure instant(GCI) is one of the important problems from speech analysis including pitch synchronous analysis of speech and estimation of source characteristics speech from voiced segments.Many different kinds of GCI detectors was suggested upto now. Many of the GCI detectors are based primarily on the linear predictive errors of speech signals. The main idea of such methods is that such error signal can indicate the most probable position of GCI. Also some improvements were made to emphasize the positions using a hilbert transform. But there are some cases that error signal cannot indicate GCI position properly in such cases as high vowels. Also those methods require additional post processing to smooth the output tracks or to remove faulty detections.In this paper we suggest a method which combines the linear prediction based GCI detectors and smoothing algorithms. As a result we can reduce the amounts of computation for the processing and can improve the performance of GCI detectors..2. GCI DETECTION METHODSThere are many methods suggested about glottal closure instant detection. Most of them are based on the error signal of linear prediction. Other methods are based on maximum likelihood or laryngograph signal which is measured directly from the outside of human vocal fold.Linear prediction based methods were EFLPR proposed by Yegnanaryana et. al. The method is based on the fact that prediction error signal is big at the location of glottal closures. This method works quite well. Generally it works for vowels or voiced consonants. But it sometimes can not show clear positions near probable GCI in such cases as high vowels or when the frequency of first formant and pitch frequency is very close or the same. So the hilbert transform was used to emphasize the position. And some post processing should be applied to adjust the delays of position.Maximumlikelihood method was proposed by O’Shaughnessy et.al. This method is based on epoch filtering theory which is used for radar signal detection. This method also used the hilbert transformation to get emphasized position signal. And this method also shows some delays in positions.Also methods using laryngograph are suggested but it is thought an inconvenient method due to attaching electrodes on the neck.3. IMPROVED METHODIn order to improve the performance of detection, we suggest additional processings to linear prediction based GCI detectors. Overall structure is shown on figure 1. The main differences with current method is in standard pitch scheme and change of S/V/UV decision stage.3. 1.Standard Pitch SchemeWe adopted standard pitch scheme to get the successive GCI locations from the speech signal. This scheme is based on the fact that the length of pitch period does not change drastically during the single segment of speech. In normal speaking conditions pitch periods do not double or halve during the limited length of the segment. So we obtained standard pitch candidate from the centerportion of the speech segment. Speech segment is decided from the scheme which will be mentioned at the next section.By using this scheme we can get interpolation effect during the computation of GCI at the same time. Also we can reduce the amount of computations by reducing the size ofthe analysis frame size.This scheme is described in figure 2.Figure2: Standard Pitch Concept3.2.S/V/UV DecisionSilence, voiced and unvoiced decisions are made with simple parameters such as zero crossing rates and energy. Normally in conventional GCI detectors they use GCI detector output to make a S,V,UV decision. The reason why we made thedecision separately is that wrong decisions for GCI can lead to wrong decisions of S,V,UV. Main errors about S,V,UV decision occurs at the initial or final stages of sounds at which the energy levels are comparatively smaller than at the stationary parts of the speech signal. We think that error rate when deciding S,V,UV from the output of GCI detector will be similar to that when deciding from the energy or zero crossings of the speech signal.By deciding voiced segments of speech separately we can reduce the computational amounts.3.3.GCI Detection AlgorithmWe made decision from the MHEWLPR(Modified Hilbert Envelope from Windowed Linear Prediction Residual). From the voiced segements of the signal we get the linear prediction residual. Then from the center part of its spectrum the normalized hilbert envelope is obtained. On most stationary voiced segments we can get quite a good GCI pointers. But for some erroneous cases such as high vowels we often fails to find the location or just find false locations. The main reasons for that is the error signal is not enough to indicate GCI position in that signal. So our standard pitch scheme is proposed to help find such ambiguous positions. Applying the standard scheme is as follows.First, get the standard pitch value from the stationary center part of the voiced segments using autocorrelation function. In this case the frame length is chosen long enough to get an averaged pitch value. Using this value as a reference we start linear prediction analysis from the initial part of the voiced segment.The first GCI candidate is aquired based on conventional method.From the second GCI position we apply standard pitch scheme.By comparing previously obtained standard pitch with the new location candidate, we can get most probable location based on the fact that the human pitch period cannot change drastically due to the relatively slow movement of the physical organs. Also we apply a hanning window before analysis to get more emphasized GCI location. This makes finding GCI location easier.Next, we remove any false locations from the forward andbackward corrections. This scheme works as follows. First we start from the center part of the segment. By comparingFigure 1: Schematic Diagram of Methodcontiguous GCI positions and pitch intervals, we remove any provable halving or doubling between 80 and 120 % of standard pitch value. In figure 3 resulting corrected MHEWLPR and GCI indicator signal is shown withcomparison to DEGG signal.Figure 3: Analysed Results from Initial /a/ sound(a) original speech (b) MHEWLPR (c) Corrected MHEWLPR (d) GCI indicator (e) DEGG4. EXPERIMENTAL RESULTSUsing the suggested algorithm we can obtain improved GCI locations from various speech materials. Then we compared the performance of the suggested method’s result with the conventional EFLPR method’s. We also compared the result with GCI locations from the differentiated laryngograph signal as a reference.To prepare speech materials speech and laryngograph signal are sampled with 11 KHz sampling frequency. To sample speech we used laryngograph processor with DSP. The analysis is done with a SPARC-10 workstation using MATLAB.List of speech samples are 10 pronounced korean numbers and Korean version of ‘The North Wind and The Sun’ and /Chang Won Dae Hak Gyo/ which means ‘Changwon National University’ in Korean.Figure 4 shows comparisons of pitch trajectories from laryngograph output and suggested method. Pitch tracks follow closely to manually detected one. At the starting points and ending points it fails to find tracks. This is caused by false detection of speech segments from energy and zero crossings. But considering the fact that laryngograph itselfoften fails to find such positions, it is quite a natural phenomena.Figure 4: Speech /Chang Won Dae Hak Gyo/ and Pitch Tracks (a) Original Speech (b) Spectrogram ( c) Pitch Track fromLaryngograph Signal (d) Pitch Track from Suggested Method Figure 5 is comparisons among 6 kinds of signals. Original Speech, LPC residual, EFLPR, MLED, suggested method, DEGG are compared. From the figures we can see that our method indicates much explicit peak positions even at the initial transient part of the signal. The other method’s output shows ambiguity at the transient parts.Table 1. Shows average non-GCI detection error rate and average false GCI detection rates for both EFLPR and our method for 5korean vowel signals. At table 1 we can see that our method gives better result and the result is much improved one from previous EFLPR method. From this result we can conclude that suggested successive method can help improve the performance of a conventional linear prediction based GCI detector.EFLPRMHEWLPR Non-GCI rate 16.02 3.4False GCI rate6.823.3Table 1. Average Non-GCI detection and false GCI detection rates for two methodsMost of the delays are concentrated on under 4 points delay. Figure 6 shows number of delay points for test materials used.5. CONCLUSIONSIn this paper we suggested an improvements to conventional linear prediction based GCI detectors. From the results we can verify that the suggested method improves the performance of detecting GCI positions compared to the conventional linear prediction based method. This method is considered to be a good one for automatic GCI detections with implicit interpolation andcorrection capability.rwsrswtrtwuruwr t v x z sr§®£»b ±«°¶µjgkFigure 6. Delay points for test materials6. REFERENCES1. T.V.Anathapadmanabha, B.Yengnanaryana,”Epochextraction from linear prediction residual foridentification and close glottis interval”,IEEE Trans.ASSP, Vol.27,No.4,pp.309-319,Aug.19792. Yang Ming Cheng, D.Oshaughness,”Automatic andReliable Estimation of Glottal Closure Instant andPeriod”,IEEE Tans ASSP,Vol.37,No.12,Dec. 19893. Ho-Gyun Bang, Cheol-Woo Jo,”A Study on theDetection of Glottal Closure Instant Using SequentialLinear Prediction”,Proceedings of KSPC’94, Vol.7,19944. A.K.Krishnamurthy, D.G.Childers , ” Two- ChannelSpeech Analysis ”, IEEE Trans ASSP , Vol.34 , No.4, pp.730 - 743, Aug.19865. D.Y.Wong,J.D.Markel,A.H.Gray,Jr.,”Least squaresglottal inverse filtering from the acoustic speechwaveform”, IEEE Trans. ASSP, Vol.27, pp.350-355,Aug. 1979Figure 5. Comparisons of GCI signalsFrom above Original Speech , Residual Signal, EFLPR,MLED,MHEWLPR,DEGG。