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Home » California Engineering Schools
California Engineering Schools
Engineering Programs in California
California's cheapest Engineering Program is located at Sacramento City College, and costs
$1,104 annually.
More students applied to University of California-Los Angeles's Engineering Program than any other school in California, with
61,556 total applicants.
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Engineering Schools in California:
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The state of California is very devoted to engineering, as it has three of its graduate schools named in the Top Five Best Grad Schools in the U.S. according to U.S. News and World Report. Because of this the U.S. Bureau of Labor Statistics places California in the top five for agricultural engineers, chemical engineers and mechanical engineers, with aerospace engineers growing at the highest employment rate of 20.4 percent.
Here are some of the top companies in California that hire engineering professionals: Chevron, McKesson, Wells Fargo, Safeway, Intel, Walt Disney, Ingram Micro, Northrop Grumman, Cisco Systems, Countrywide Financial, Occidental Petroleum, Gap, DIRECTV Group, Computer Sciences, Amgen, Sun Microsystems, Health Net, Edison International, and Wal-Mart.
Whether a student chooses to attend one of the many state colleges or universities or one of the private ones, he or she will find a wide range of engineering degrees. Most colleges offer associate, bachelor, master or doctorate degrees, although others offer additional certificates. Some students get their associate degrees at two-year colleges, such as American River College, and transfer to the larger ones to get higher degrees.
California Institute of Technology is one of the private, not-for-profit colleges that have degrees in several of the most popular engineering fields, including chemical engineering and civil engineering. California Polytechnic State Universities are some of the best schools in the country for engineering students.
Outlook for Engineering Jobs in California
In general, California is a major job market for engineers. The state's diverse range of industries is evident in its employment record with the Bureau of Labor Statistics (BLS). Although the BLS predicts a slow to average job growth rate for engineers, some states will see an above average job growth. According to a 2012 BLS report, California has the highest employment rate for aerospace engineers, biomedical engineers, civil engineers, mining and geological engineers and others.
Programs in California


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Sheldon Lee Glashow
1979 Nobel Prize in Physics
American theoretical physicist, the Metcalf Professor of Mathematics and Physics at Boston University and
Eugene Higgins Professor of Physics, Emeritus, at Harvard University, and the Advisory Committee President of Meso-American Institute for Theoretical Physics. He shared the 1979 Nobel Prize in Physics with Steven Weinberg "for their contributions to the theory of the unified weak and electromagnetic interaction between elementary particles, including, inter alia, the prediction of the weak neutral current"
1954, BA from Cornell University
1958, Ph.D. from Harvard University
1966-Present, Professor, Higgins Prof. of Physics, Emeritus, Harvard University
2000-Present, University Professor, Boston University
1979, Nobel Prize in Physics
Member of the National Academy of Sciences
Member of the American Academy of Arts and Sciences Member of the American Physical Society
In 1961, Glashow extended electroweak unification models due to Schwinger by including a short-range neutral current, the Z0. The resulting symmetry structure that Glashow proposed, SU (2) × U (1), forms the basis of the accepted theory of the electroweak interactions. For this discovery, Glashow along with Steven Weinberg and Abdus Salam, was awarded the 1979 Nobel Prize in Physics.
In 1964, in collaboration with James Bjorken, Glashow was the first to predict a fourth quark, the charm quark. The prediction of the charm quark also removed a technical disaster for any quantum field theory with unequal numbers of quarks and leptons. In 1973, Glashow and Howard Georgi proposed the first grand unified theory. This work was the foundation for all future unifying work.

redpajama_set_name: RedPajamaCommonCrawl


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Fresh blow for greenhouse sceptics
By Gabriel Walker
THE RISE in global temperatures over the past century was caused mainly by increased carbon dioxide emissions, according to an analysis of temperature records by David Thomson of AT&T Bell Laboratories in Murray Hill, New Jersey. Thomson hopes his study will silence the sceptics who argue that the observed 0.6 °C warming has nothing to do with the greenhouse effect, and was instead caused by an increase in the radiation coming from the Sun.
Thomson has reanalysed historical temperature records to see whether solar variability could have warmed the world over the course of this century. He reasoned that if the Sun is getting brighter, the amplitude of the seasonal temperature cycle – the temperature difference between winter and summer – should be increasing. This is because a disproportionate amount of the radiation reaching the Earth from the Sun ends up heating those parts of the planet that are experiencing summer.
But when Thomson looked at a temperature record compiled from land and sea observations throughout the northern hemisphere over the past 140 years, he found that since about 1920 the amplitude of the annual temperature cycle has actually decreased (Science, vol 268, p 59). So increased solar radiation cannot be responsible for the recent warming trend. Indeed, the new analysis may be the best evidence yet that greenhouse warming has begun, as CO2 should have a disproportionate effect on winter temperatures, reducing the amplitude of the seasonal temperature cycle. This is because CO2 in the atmosphere acts like a blanket, preventing the Earth's heat being lost to space. This effect is more important in winter than in summer.
Thomson claims that increased CO2 emissions have also altered the timing of the seasons. Until the early part of this century, he discovered, the seasons had tended to start later and later. This is because the calendar counts a year as the time it takes to complete a regular cycle of varying length of day versus night. But this cycle is about 20 minutes shorter than the time it takes for the Earth to orbit the Sun. It turns out that it is the Earth's proximity to the Sun – which depends on its position in its elliptical orbit – that dictates the timing of the seasons, so they should arrive slightly later each calendar year.
After 1940, however, Thomson found that there was a sudden dramatic shift. In the northern hemisphere the seasons suddenly started arriving earlier, rather than later, and now they are back to their 1850 timing. Thomson points out that this shift coincided with an acceleration in CO2 emissions. The size of this shift fits exactly with the magnitude of the greenhouse effect expected from increased CO2 emissions. Thomson also found a close correlation between the amount of CO2 in the atmosphere and the average temperature increase since 1940.
Thomson believes that these results finger CO2 as the culprit for the recent warming, although it is not yet clear precisely how increased emissions of the gas have altered the timing of the seasons. Just to be sure that other factors are not involved, he also looked at other phenomena that could affect climate, such as volcanic eruptions and El Niño reversals in the warm currents in the Pacific Ocean, to see if they could explain any of the changes. Although he found short-term correlations between these factors and the temperature record, only CO2 could explain the long-term warming trend. "You almost have to invoke magic if it's not CO2," he says. "It's the only logical explanation."
Magazine issue 1974 , published 22 April 1995
Have the stars being lying about their age?
Tadpoles snatch savlation from the jaws of disaster


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in Postage Stamps
by Hugo Vargas
This year marks the 100th anniversary of the death of the Nobel Prize benefactor, Alfred B. Nobel (1833-1896). A chemical engineer and scientist by profession, Nobel devoted most of his life's work to the research and development of explosives, specifically dynamite, nitroglycerin, and blasting gelatin. Eventually, he came to hold 355 patents and became incredibly wealthy from the exploitation of his inventions-building and operating over 90 factories in 20 countries on five continents.
A significant portion of funding for the Prize can also be traced to Azerbaijan. Alfred's brothers, Robert and Ludvig, owned the Nobel Brothers' Petroleum Company at the time when Baku was producing more than 50% of the world's oil (See AI: 2:4, 10; Summer 1994). As Alfred was the largest single shareholder in the company, it is said that the decision to allow the withdrawal of his shares from Baku (a reported 12% of the Prize Fund) enabled the awards to be established.
Since 1901, prizes have been awarded annually in the fields of physics, chemistry, physiology or medicine, literature and peace. The economics prize was added in 1968. Each Prize consists of a gold medal, a diploma bearing a citation, and approximately $1 million.
Unlike other contemporary barons of business, Nobel lived a rather spartan life. He is said to have been unassuming and reticent. Though Swedish by birth, his outlook was cosmopolitan, and he felt comfortable with French, German, Russian, English and Swedish languages. The scope of the prizes reflect his own broad personal interests.
Nobel's great legacy to the development of intellectual thought and world peace has not gone unnoticed among the world of philatelists. More than 175 stamps have been issued over the years from 15 countries, honoring Nobel Prize winners as well as Nobel himself on the anniversaries of his birth (1833), death (1896), and the creation of his will (1895).
Announcements of the Nobel winners are made early each October, followed by the Award Ceremonies on December 10, in Oslo (Peace) and Stockholm (remaining prizes).
So far, 15 countries have celebrated Nobel's legacy with 175 stamps, most of which honor Prize recipients of the last 95 years. Others commemorate Nobel's birth or death anniversaries.
Other articles related to the Nobel Brothers:
(1) Nobel Prize Funded from Baku by Adil Baghirov (AI 4:2, Summer 1996)
(2) Toni Morrison's Nobel Prize Acceptance Speech, (AI 6.3, Autumn 1998)
(3) Newsmakers: Nobel Prize Centennial by Brita Asbrink (AI 9:3, Autumn 2001)
(4) Nobels in Baku: Swedes' Role in Baku's First Oil Boom


Celebrating 25 years of existence, but also the continuity of said existence, the "best of" setlist they've performed featured a new song, "The Lightning-Struck Tower".
Gallery | This entry was posted in Concerts and tagged 25 years, anniversary gig, dark metal, gothic metal, hard club, Heavenwood, raising legends. Bookmark the permalink.


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Stratford police shoot man they say opened fire on officers
STRATFORD >> Connecticut State Police say an armed man shot by Stratford police fired at least one round at officers first.
Trooper Kelly Grant says city officers responded to a waterfront section of town known as Lordship at about 4:45 a.m. Wednesday for reports of a man with a gun.
Officers found the man and ordered him to drop the weapon. Grant says the man advanced toward officers and fired at least one round. At least one Stratford officer returned fire and struck the man.
The suspect was given first aid at the scene before being taken to Bridgeport Hospital. His condition was not disclosed.
No names were released.
No additional information was made public.
State police are investigating.


The segment is actually a remix of this video made in collaboration with a fine arts college, in which different students got to direct the actor in front of a greenscreen, and release the segments later under a Creative Commons license. So edit away, internet. This TED video is only the beginning.


54mm work covers scales from about 1/28th to 1/35th and there are examples of houses, hotels, landscapes and dioramas.
I started taking this size commissions from when we started selling the John Jenkins figures in about 2006/7. 2 years later and a large proportion of my work is now in 54mm scale. It is a very rewarding scale to work in as the size allows more detail in the model and the customer tends to get alot of 'bang for their buck'.
Landscape commissions are probably the most common. I can do a simple grass board that can suite any period and can be done in any size right up to complicated snowy landscapes with a river and log cabin. I can design them so they will fit a number of periods.
More examples of dioramas and displays include shelf sized displays. These are on 18mm thick MDF bases. It makes an excellent foundation for whatever I build on it and can easily be cut to any size you want. These four examples are all on 2ft long by 1 foot wide boards.
Commissions for architecture vary from the individual ruin to large hotels,churches and historical buildings.
These last pictures of the Normandy town are from Wargames Illustrated magazine from a photo shoot with them in 2009.
The figures are mainly Collectors Showcase WW2 with the odd plastic one painted by me. All the vehicles are by Collectors Showcase.


The team "Chimie Théorique, Méthodologies, Modélisation" belongs to the « Chemical Physics » Department of the ICGM.
All members of CTMM are theoreticians or computational chemists using quantum approaches in their research. The aim of CTMM is to develop methods and concepts to analyze the physico-chemical properties and the chemical transformations occuring in molecular chemical systems and materials. The topics covered span a large spectrum from small molecular quantum dynamics, to the structure and reactivity of transition metal complexes, and to the electronic properties of organic and inorganic solids. All the theoretical studies are carried out in synergy with experimental groups.
The research activities in CTMM are articulated around two axes : « Methodological Developments » and « Applications ». The first research theme is concerned by formal, conceptual, algorithmic or computational developments of quantum methods and methodologies able to treat complex systems (molecules, solids, interfaces) and their environment . The second research theme deals with high societal impact application in various domains such as « Physico-chemistry of the environment », « Homogeneous and heterogeneous catalysis » and « Materials for Energy ». This synergy between Developments and Applications is the cornerstone of the research activities carried out in the team and transcends the particularities of each problems adressed in order to reach a better understanding of chemical reactivity.
The CTMM team originates from the integration in January 2007 in the Institut Charles Gerhardt of the former UMR 5636 (Laboratoire de Structure et Dynamique des Systèmes Moléculaires et Solides, LSDSMS) created in 1995. The LSDSMS laboratory was created around a nucleus of people coming from the Theoretical Chemistry Laboratory in Orsay with the aim to develop theoretical chemistry in Montpellier. In the last years, the CTMM team has seen its composition changed with the departure of 3 senior members (Odile Eisenstein, Claude Leforestier and Gérard Parlant) and the hiring of two young staff members (Thibaud Etienne (lecturer) and Matthieu Saubanère (research-associate)). These changes have resulted in a reorganisation of the research activities in the team around a common methodological goal. This emerging theme relies on the expertises in electronic structure and vibrational structure calculations available in the laboratory. It aims at reaching a better description of the physico-chemical properties in various applications related to energy conversion, transfer and storage.
The "Chimie Théorique, Méthodologies, Modélisation" Team is located in the University of Montpellier, Faculty of Sciences ( building 15 , RdC).


Rochester, MN (KROC-AM News) - The extended winter is apparently having an impact on new home construction in Rochester. The City Building Safety Department issued only 25 permits for new single-family homes last month. It was the lowest total for the month of March since 2011. The city has now issued 60 permits for new single-family homes since the start of the year, which is almost 20 below the total through the first three months of last year.
Overall construction activity was strong last month. The city issued 165 building permits valued at $47.3 million. That pushed the total for the year to over $113 million, which is almost $40 million above the total through the first quarter of 2017.
Over half of the valuation total last month was due to one project. The Mayo Clinic took out a permit worth almost $29.8 million for the ongoing expansion of the Generose Building at St. Marys Hospital. The second largest permit was for the new Altra Credit Union location in northwest Rochester that was valued at just over $4.1 million.


TOP's Crumplets for small parrots and ones who don't like to hold their food in their foots.
Certified Organic Ingredients: Rice, hulled millet, sunflower seed hulled, sesame seeds unhulled, quinoa whole, buckwheat hulled, dandelion leaf powder, carrot powder, spinach leaf powder, purple dulse, kelp, rose hips powder, rose hips crushed, orange peel powder, lemon peel powder, rosemary whole leaf, cayenne ground, crushed red chili peppers, nettle leaf. Non certified organic, ecologically, sustainable ingredient: barley, alfalfa leaf.
TOP's Crumplets do not contain any fillers, like soy or corn.NO BHA, BHT or ethoxyquin. NO artificial colors, flavors, vitamins, preservatives, or sucrose/sugar. No wheat or any nuts.


Should I go for Android app development or for iPhone app development? It is one of the most confusing questions which never actually get answered. With all of the blogs and articles that are circulating on the topic, you might be thinking that mobile app world would have come to some conclusion by now on iPhone and Android app development. But unfortunately, it's yet to be concluded. And, as there is no specific answer to the question — both platforms are great, for the same and diverse reasons.
We created AppSquadz because we know that there are better ways to develop mobile apps. We at AppSquadz know is the need of time – A modern programming language, dominant tools, a proficient and wonderful workflow and the command to craft the best user experiences. We dedicate ourselves to make the process fast, easy and fun to develop innovative mobile apps.
Whether you are looking for a custom, native Android, iOS, Windows or Blackberry app? Or looking for a cross-platform responsive app? If you want it, we can create it. Where others fail, we excel.
Looking to Develop Android App?
Android apps run in a highly scattered multi-device world. Although API rules are flexible, but quality assurance inspection is more rigorous. For AppSquadz it's not a new thing. We've developed hundreds of amazing Android apps on releases from Kitkat to Marshmallow, from simple phones to custom devices — we've experienced it all.
Looking to Develop IPhone App?
Having successfully developed and delivered hundred of iPhone App Development Projects, we are on the verge of becoming top app development company. Our squad is adept in designing and developing apps for iPhone 4 or 4s, iPhone 5 or 5s, iPhone 5c, iPhone 6 and iPhone 6 Plus. Soon we will be optimizing several apps for iOS 7, iOS 8 and iOS 9 Beta 2 as well.
We are a Spartan of mobile application development. We have worked with top brands and we never settle for just "good" and neither should you when it's about hiring a mobile app development company. For last two years, AppSquadz has been crafting remarkable apps for some of the world's most renowned brands. Our 100+ team members comprise industry specialise in UI/UX design, mobile app, and marketing strategy. We are not only iPhone and Android app development service provider, we are also capable of developing apps for Windows and Blackberry based mobile operating system.
We at AppSquadz believe that mobile application should not only be innovative but should also provide an amazing user experience which users will remember forever.
Internet Of Things and other Trending Technologies: How are they Connected?


Do come down to The Farm pub in sunny (we hope) Bristol this Saturday 28th May for a right shindig with the Pigpen Social lads n lasses.
There's a host of live acts, including a choir, a looping drum and sound chap, funk bands, hip hop artists and the like, seamlessly segue'd by yours truly, outdoors in the extensive garden.
Every time I've been asked to play I've always had a wicked time, so if you fancy a music filled saturday afternoon and evening, may I suggest popping down.
It's free, but it IS for Oxfam so please do lob some change in the buckets if you're suitably entertained.


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CURIOUS INDEX, 9/27/07
By Orson Sep 27, 2007, 10:42am EDT
Share All sharing options for: CURIOUS INDEX, 9/27/07
Oh, yes. He's a wideout. Mario Manningham celebrated two wins in a row for Michigan by doing the worm. No extra sauce needed for this:
(HT: The Wiz) The Wiz speculates that the worm was popularized by the wrestler Scotty 2 Hotty, which may be true in the short run; however, the Wikipedia entry credits the move to Sophie Tucker, a '20s vaudevillian with a voracious sexual appetite and who, in her old age, looked like Ralph Friedgen in drag. Our world spins somewhat more eccentrically knowing this.
Pat Sims, public relations genius. Auburn defensive tackle, he of the complete and total stoppage of Deshawn Wynn on the goal line versus Florida last year, may have been "light-hearted (and)... not boastful" when he said this to the media this week about the cast he wears on his hand to protect his fingers.
"Hes not going to run through me," Sims said. "When he gets a feel of this club he isn't going to want more problems."
Whew, that's piquant! Practically Oscar Wilde-ish! Were Florida that witty, we'd have players saying things like "I'm going to hit Brandon Cox so hard he shatters into six individual American Girl dolls." Just light-hearted fun like that--not boastful, or calling advance attention to something I might swing at an opposing qbs head, balls, knees, or throat, raising the risk of a personal foul and a loss of fifteen yards for my team.
He's really better at center. Or wideout. Or anywhere, really. The nation's leader in sacks isn't playing at the position Jim Leavitt wanted him to play at--center. USF's George Selvie has 8.5 sacks on the year already, and could have a few more given Pat White's 300 carries a game.
In case you were wondering, that's no typo: each person in the West Virginia backfield carries the ball 300 times a game. Rich Rodriguez doesn't just control the line of scrimmage--he's got wormholes and temporal distortions on his side. That's the kind of shit the spread-option does to the fabric of the universe, lawya.
A hero named Swindle. Not us--Ken Swindle, the Tuscaloosa police chief whose department who nabbed three UPS employees stealing tickets from Alabama season ticket holders through the mail and routing them to ticket brokers. Unstoppable men, the Swindles.
You want hell? She's bringing it with her.
Someone call William Proxmire! He's dead? Get him anyway! The highest paid employee of the federal government is not in the executive branch, but rather in the Navy: their football coach Paul Johnson, who makes a million a year, more than the President, VP, Secretary of State, and that brave soldier who swims five miles at night underwater to slap a mine on the side of a ship, crawls ashore, makes love to a beautiful woman, rescues her, and flies off with the blueprints in an enemy helicopter just as the harbor explodes. Their name? Well, her enemies know her as the Black Widow, but around the White House, they just call her Secretary of Labor Elaine Chao, the best soldier this army's got, friend.
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BioMed Research International
BioMed Research International / 2012 / Article
AbstractIntroductionMethodsResultsDiscussionConclusionDisclosureAcknowledgmentsSupplementary MaterialsReferencesCopyright
Deepak Poudyal, Xiangli Cui, Phuong Mai Le, Tia Davis, Anne B. Hofseth, Yu Jin, Alexander A. Chumanevich, Michael J. Wargovich, Mitzi Nagarkatti, Prakash S. Nagarkatti, Anthony Windust, Lorne J. Hofseth, "A Limited Role of p53 on the Ability of a Hexane Fraction of American Ginseng to Suppress Mouse Colitis", BioMed Research International, vol. 2012, Article ID 785739, 11 pages, 2012. https://doi.org/10.1155/2012/785739
A Limited Role of p53 on the Ability of a Hexane Fraction of American Ginseng to Suppress Mouse Colitis
Deepak Poudyal,1 Xiangli Cui,1 Phuong Mai Le,2 Tia Davis,3 Anne B. Hofseth,1 Yu Jin,1 Alexander A. Chumanevich,1 Michael J. Wargovich,4 Mitzi Nagarkatti,5 Prakash S. Nagarkatti,5 Anthony Windust,2 and Lorne J. Hofseth1
1Department of Pharmaceutical and Biomedical Sciences, South Carolina College of Pharmacy, University of South Carolina and Medical University of South Carolina, Columbia, SC 29208, USA
2Institute for National Measurement Standards, National Research Council, Ottawa, ON, Canada
3Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, USA
4Department of Cell and Molecular Pharmacology & Experimental Therapeutics and Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29245, USA
5School of Medicine, University of South Carolina, Columbia, SC 29208, USA
Academic Editor: Paul W. Doetsch
Revised25 May 2012
Accepted11 Jun 2012
Ulcerative colitis (UC) is debilitating and carries a high colon cancer risk. Apoptosis of inflammatory cells is a key mechanism regulating UC. We have recently shown that American ginseng (AG), and to a greater extent, a Hexane fraction of AG (HAG) can cause apoptosis and suppress mouse colitis through a p53-mediated mechanism. Here, we tested the hypothesis that HAG suppresses colitis through a p53 mechanism. We found only a limited impact of p53 in the ability of HAG to induce inflammatory cell apoptosis and suppress mouse colitis in vitro and in vivo. Finally, we asked whether HAG could cause cell cycle arrest of HCT116 colon cancer cells in vitro. Interestingly, HAG caused a G1 arrest of such cells independent of p53 status. Findings are significant because HAG suppresses colitis and associated colon cancer, and mutation in p53 is observed in most colitis-driven colon cancers. Therefore, HAG might be very effective in targeting the inflammatory cells and cancer cells since it induces apoptosis of inflammatory cells and cell cycle arrest in both p53−/− and WT p53 colon cancer cells.
Inflammatory Bowel Disease (IBD) is considered an autoimmune disease that causes chronic inflammation in the gastro-intestinal tract. Ulcerative colitis (UC) is one of the IBDs' reflecting chronic relapsing inflammatory disorders of the intestine [1]. Among the cell types, epithelial cells, myeloid innate cells, effector T cells, regulatory cells, and B cells have been implicated in IBD pathogenesis [2]. Excessive effector T-cell cytokine secretion or defective regulatory T cells lead to the disease propagation of IBD [2]. The intestinal barrier performs two important tasks to keep the balance between health and disease. First, it must mediate effective absorption of fluids, nutrients, and minerals from the lumen across the epithelium and into the microcirculation and microvilli [3]. Second, the barrier must be impermeable to prevent the transfer of potentially pathogenic microbes and infectious agents [3]. Dysfunction in this intestinal barrier leads to the dysfunction in the intestinal immune system, which has been implicated as the major mechanism by which chronic inflammation occurs in colitis [4]. Hence a common feature of IBD pathogenesis is the dysregulated effector T cell response to commensal microbiota. Because the intestinal mucosa is constantly exposed to several antigens, the mucosal immune system have evolved several strategies to avoid an unnecessary and uncontrolled inflammatory reaction. Once the antigen from the commensal microbiota has been eradicated, T lymphocytes of the intestinal mucosa require a method to attenuate the local immune response [4]. One of the regulatory methods, downregulation of activated T lymphocytes via apoptosis, is a very potent and effective strategy, now considered as a key controlling mechanism of IBD [5]. Failure to regulate T-cell responses in the intestinal or colonic mucosa leads to an inappropriate and sustained injurious immunologic reaction [6, 7]. Because of their heightened activation and activity in IBD pathogenesis, effector T cells are considered an excellent target for therapeutics.
Cells undergoing apoptosis are characterized by the shrinkage and condensation of their cytoplasm, increased mitochondrial permeability, chromatin condensation into caps at the edge of the nucleus, DNA fragmentation, and the appearance of plasma membrane blebs, often referred to as "apoptotic bodies" [8]. There are two signaling pathways for the cell death by apoptosis: the intrinsic and extrinsic pathways [9, 10]. Activation of the extrinsic pathway occurs by ligand-induced cell surface receptor (e.g., tumor necrosis factor receptor 1 TNFR1, Fas, and death receptor 5) activation [11]. The intrinsic pathway is activated with growth factor deprivation, oncogene activation, or when DNA damage is detected by cellular sensors such as ataxia telangiectasia mutated (ATM), ataxia telangiectasia and Rad3-related (ATR), and tumor protein 53 (p53) [11]. Inflammation-induced reactive oxygen species and nitric oxide lead to p53 stabilization and accumulation [12, 13]. This activates p53 to eliminate the damaged cells by apoptosis [12, 13]. This is an intrinsic approach for the apoptosis of damaged cells, where activated p53 plays an important role in mediating apoptosis in epithelial and inflammatory cells during the process of colitis [4, 12, 14–17]. The post-translational modification (often phosphorylation at Serine 15) tends to elevate the level of wild-type (WT) p53 during inflammation [12]. p53 plays at least 2 separate roles in the responses to therapeutic agents It is an important component of cellular checkpoints (cell cycle arrest), and it can mediate apoptosis [18]. The role of p53 in the responses of tumor cells to therapy is controversial. For example, loss of p53 function can cause resistance to 5-fluorouracil (5-FU), but increased sensitivity to DNA damaging agent such as Adriamycin [18]. This provides a clear indication that some drugs may exert an apoptosis-inducing effect through a p53-dependent while other drugs effect through a p53-independent pathway.
Cancer is one of the scenarios where too little apoptosis occurs, resulting in malignant cells that will not die and continue to proliferate. Four cellular functions are inappropriately regulated in the cancer cells: cellular proliferation, differentiation, chromosomal and genetic organization, and apoptosis (Reviewed in [19]). Abnormal cell proliferation leading to accumulation of clonal cells is seen in cancer. As mentioned this could be due to the defects with the cell cycle control. Upon sensing DNA damage, p53 is activated, resulting in either a G1 cell cycle arrest [20, 21] or apoptosis [22, 23]. Cells undergo a G1 cell cycle arrest to allow the DNA repair before replication and if the DNA damage is beyond repair, the cells tend to undergo apoptosis. Cells can also undergo a G1 arrest due to targeting cyclins and cyclin-dependent kinases. p53 mutation is observed in most cancers [24], which often tend to relax this G1-S cell cycle transition because p53 could not be activated. Hence, cancer cells lack the appropriate G1-S checkpoint regulation and controlled apoptosis.
American ginseng (AG, Panax quinquefolius) is grown in the eastern temperate forest areas of North America, from British Columbia, southern Quebec, Ontario, Minnesota, and Wisconsin in the north, to Oklahoma, the Ozark Plateau, and Georgia in the south [25]. AG is an obligate shade perennial native of North America and its root is the commonly used component. AG has been reported to have a wide range of pharmacological effects, including effects on the central nervous system, blood-sugar levels, cardiovascular system, endocrine system, immune system, and cancer [4, 26, 27]. Recently we have shown AG extract suppresses colitis by the accumulation and activation of p53 to induce apoptosis of inflammatory cells [4]. To further delineate the active anti-inflammatory and proapoptotic components present in the AG, we sub-fractionated a different fraction of AG. A Hexane Fraction of AG (HAG) showed increased anti-inflammatory and pro-apoptotic properties in the chemically induced mouse model of colitis [28]. The elevation of WT p53 levels during inflammation [12] resulting in apoptosis of inflammatory and damaged cells [4, 29, 30] led us to the notion that the active anti-inflammatory components present in HAG might suppress colitis through the p53 pathway. Here, we tested this hypothesis.
2. Material and Methods
2.1. Bioassay-Guided Fractionation of Hexane Fraction of AG (HAG)
The P. quinquefolius extract has been described previously in detail by our laboratory [16]. As well, we have recently described the generation of the HAG used in the present study [28].
2.2. Chemicals and Reagents
Dextran sulfate sodium (DSS) was purchased from MP Biomedicals (Solon, OH: molecular weight, 36,000–50,000).
2.3. Cell Culture and Treatment
TK6 (p53+/+) and NH32 (p53−/−) cell lines were a kind gift from Curtis Harris (National Cancer Institute), originally derived from Dr. William Thilly's and Howard Liber's labs. TK6 cells are a lymphoblastoid cell line derived from the spleen more than 30 years ago [31]. NH32 cells are an isogenic derivative of TK6 cells in which both alleles of the p53 gene were knocked out [32]. Jurkat T cells are an immortalized line of T lymphocyte cells derived in the late 1970s from the peripheral blood of a 14-year-old male with T-cell leukemia [33]. Jurkat T cells have a defective p53 pathway due to a mutation in the COOH-terminal domain responsible for transactivation [34, 35]. TK6, NH32, and Jurkat T cells were maintained in exponentially growing suspension culture at 37°C in a humidified 5% CO2 atmosphere in RPMI 1640 supplemented with 10% heat-inactivated calf serum, 100 units/mL penicillin, 100 μg/mL streptomycin, and 2 mmol/L l-glutamine.
CD4+/CD25− cells from C57BL/6 mice were purified from the spleens using nylon wool columns (Polysciences, Warrington, PA, USA) followed by depletion of B cells and macrophages. CD4+/CD25− T cells were then isolated using an MACS miniseparator and CD4 and CD25 microbeads according to the manufacturer's instructions (Miltenyi Biotec, Auburn, CA, USA) by depletion of CD4−CD25+ T cells (negative selection). The purity of T cells was 88.6% (Supplementary Figure 1 available online at http://dx.doi.org/10.1155/2012/785739) as determined by flow cytometry (Cytomics FC 500, Beckman Coulter, Brea, CA, USA). Briefly, isolated CD4+/CD25− T cells were washed with PBS and 2 × 105 cells were re-suspended with 100 μL of PBS and incubated with anti-mouse CD4 antibody conjugated to Allophycocyanin (APC) (Clone: GK1.5; Isotype: rat IgG2b, kappa, eBioscience, San Diego, CA, USA) (0.125 μg/100 μL of cell suspension). The anti-mouse CD25 antibody conjugated to R-Phycoerythrin (PE) (Clone: 7D4; Isotype: rat IgM, Miltenyi Biotec, Auburn, CA, USA) was incubated during the isolation of CD4+/CD25− effector T cells according to the manufacturer's instructions (Miltenyi Biotec, Auburn, CA, USA). The purity of CD4+ T cells was determined by obtaining a dot plot of FL-4 (APC) versus FL-2 (PE) and individual histogram plots of FL-4 and FL-2 versus the number of events (Supplementary Figure 1). The isogenic control was the nonantibody treated isolated effector T cells.
Isolated CD4+/CD25− effector T cells (1 × 106) were cultured in six-well plates overnight followed by experimentation as indicated. All cells were maintained in exponentially growing suspension culture at 37°C in a humidified, 5% CO2 atmosphere in RPMI 1640 supplemented with 10% heat-inactivated calf serum, 100 units/mL penicillin, 100 μg/mL streptomycin, and 2 mM L-glutamine.
HCT-116, human epithelial colon cancer cells line, proficient in p53, and isogenic HCT-116 p53−/− cells were maintained in exponentially growing adherent culture at 37°C in a humidified 5% CO2 atmosphere in RPMI 1640 supplemented with 10% heat-inactivated calf serum, 100 units/mL penicillin, 100 μg/mL streptomycin, and 2 mmol/L l-glutamine.
2.4. DSS Mouse Model of Colitis
We followed our previous protocol for our DSS (MP Biomedicals, Solon, OH: 36,000–50,000 mw) mouse model of colitis [16]. Briefly (Figure 1), 8–10-week-old C57BL/6 mice received either water ad libitum or 1% DSS. All mice were on an AIN93 M diet as described previously [36]. 11.9 mg/kg of HAG was dissolved in 100 μL 1 × PBS per mouse and administered daily by oral gavage (per os, PO). 11.9 mg/kg daily is the human equivalent dose of 58 mg daily [37]. Of note, currently the use of ginseng in human clinical trials can range anywhere from 200 mg to 9 g daily [38, 39]. The control group of mice was given 100 μL of 1 × PBS by oral gavage. All procedures performed were in accordance with the Guide for the care and Use of Laboratory Animals (National Research Council, Washington, DC, USA) and approved by the Animal Resource Facility, University of South Carolina, Institutional Animal Care and Use Committee. To determine whether the HAG can prevent the onset of colitis, mice were fed 1% DSS for 2.5 cycles (7 days DSS, 7 days water making 1 cycle). The vehicle or the HAG was administered daily by oral gavage 7 days prior to the first DSS exposure and continued throughout the course of the experiment. Mice were euthanized at 2.5 cycles (Figure 1). For pathology, colon tissue samples were washed with phosphate-buffered saline (PBS; Mediatech, Herndon, VA, USA), cut longitudinally, swissrolled, then formalin-fixed overnight, and paraffin-embedded.
Experimental protocol for the DSS, prevention mouse model of colitis. 11.9 mg/kg/day of HAG or vehicle (1 × PBS) was given to the respective group of mice by oral gavage 7 days prior to the first DSS cycle and continued daily until the end of the experiment (2.5 cycles). The mice (both C57/BL6 p53−/− and C57/BL6 p53+/+) were euthanized on day 35.
2.5. Quantification of Inflammation to Examine Effects on Colitis
Paraffin-embedded tissues were serially sectioned, and one section from each mouse was stained with H&E. Sections were microscopically examined by two blinded investigators (D.P and X.C) for histopathologic changes using a scoring system previously used and validated by ourselves and others [4, 28, 40, 41]. Histology score was determined by multiplying the percent involvement for each of the three following histologic features by the percent area of involvement [4]: inflammation severity (0, none; 1, minimal; 2, moderate; 3, severe), inflammation extent (0, none; 1, mucosa; 2, mucosa and submucosa; 3, transmural), and extent of crypt damage (0, none; 1, one third of crypt damaged; 2, two thirds of crypt damaged; 3, crypts lost, surface epithelium intact; 4, crypts lost, surface epithelium lost). Percent area involvement was defined as: 0, 0%; 1, 1–25%; 2, 26–50%; 3, 51–75%; 4, 76–100%. Therefore, the minimal score is 0 and the maximal score is 40. Since, DSS-induced colitis in mice leads to the damage in colonic epithelial barrier and is characterized by extent and depth of inflammation, a grading or scoring system of inflammation with all these parameters provides an excellent measure of histologic assessment of DSS-induced colitis.
2.6. Western Blot Analysis and Antibodies
Western blots were carried out as described previously [42]. Antibodies used include: p53 (mouse monoclonal, DO-1, diluted 1 in 500, cat# OP43T; Calbiochem, Gibbstown, NJ, USA) and GAPDH (Rabbit monoclonal, diluted 1 in 1000, cat# 5174 P; Cell Signaling Technology, Danvers, MA, USA). Horseradish peroxidase-conjugated anti-mouse and anti-rabbit secondary antibodies were purchased from Amersham Biosciences (Piscataway, NJ, USA). Both secondary antibodies were diluted at 1 : 2000. All antibodies were diluted in 5% milk/PBST (0.1% Tween 20 in 1 × PBS). Western blot signal was detected by Pierce ECL Western Blotting Substrate (Thermo Scientific, Rockford, IL, USA) and developed onto Hyperfilm (GE Healthcare Life Sciences, Pittsburgh, PA, USA). Briefly, after treating blot with the chemiluminescent substrate (Pierce ECL) for a minute, the blot was exposed to the hyperfilm in the dark (Exposure time was optimized based on the band signal obtained) and the film was developed in an automatic X-ray film processor (Futura Classic E automatic X-ray film processor, Fisher Industry, Geneva, IL, USA).
2.7. Annexin V Assay
CD4+/CD25− effector T cells were seeded at 1 × 106 per well into six-well dishes for 24 h in triplicate ( 𝑛 = 3 ). Fresh medium or fresh medium containing freshly dissolved indicated concentrations of HAG (0–300 μg/mL) was added for 24 h, as indicated. Cells were then harvested for Annexin V according to instructions provided by the kit manufacturer (BD Biosciences, San Diego, CA, USA). Annexin V/propidium iodide (PI) staining was examined using a Beckman Coulter Cytomics FC500 flow cytometer.
2.8. Cell Cycle Analysis
1 × 106 cells/wells of HCT-116 WT and HCT-116 p53−/− cells were incubated in 1.0% NBCS supplemented RPMI-1640 media for 24 hrs in 6-well culture plate. The media was changed and the cells were treated with Hexane fraction of AG (0–500 μg/mL). The cells were harvested after 24 hrs of treatment and cell cycle assay was performed by labeling the dsDNA of the cells with DAPI (4′,6-diamidino-2-phenylindole) (Sigma-Aldrich, MO, USA). Briefly, the harvested cells were fixed by gently vortexing and adding 70% ethanol dropwise. The fixed cells were incubated at 4°C for at least 30 minutes. The cells were washed with PBS/1% BSA and stained with 1 μg/mL of DAPI (in PBS/0.1% Triton X-100) for 10 min. at room temperature. 20,000 cells/events were directly analyzed by BD-LSR-II flow-cytometer (BD Biosciences, San Jose, CA, USA). Based on the DNA content, the different phases of the cell cycle was determined by using BD FACSDiva software (BD Biosciences, San Jose, CA, USA). Experiments were repeated three times.
2.9. Flow-Cytometric TUNEL (Terminal Deoxynucleotidyl Transferase-Mediated dUTP Nick-End Labeling) Assay
TK6 (p53 WT), NH32 (isogenic p53−/−), and Jurkat T (dysfunctional p53) cells were incubated in 0.1% NBCS supplemented RPMI-1640 media for 24 hrs. The media was changed and the cells were treated with Hexane fraction of AG (0–500 μg/mL) as indicated in Figure 2. Cells were harvested after 24 hours of treatment and TUNEL assay was performed as described by vendor (Roche Diagnostics, IN) in triplicate ( 𝑛 = 3 ). Briefly, 1 × 1 0 6 cells were fixed using a 100 μL of fixation solution (2% paraformaldehyde) and permeabilized using a permeabilization solution (0.1% Triton X-100 in 0.1% sodium citrate). Cells were washed and incubated with TUNEL reaction mixture (label solution and enzyme solution) (Roche Diagnostics, IN, USA). Apoptosis in the samples were analyzed by flow cytometry (Beckman Coulter, CA, USA). The fluorescence was evaluated using the excitation wavelength of 488 nm and detected in the range of 515–565 nm (green, FL-1 channel). The dot plot of FS Versus FL-1 and histogram plot of (number of event) Versus (FL-1 channel) were plotted to obtain a percentage increase in the apoptosis of the Hexane fraction of AG treated cells. Positive control for apoptosis is the fixed and permeabilized cells treated with DNase I recombinant (3 U/mL in 50 mM Tris-HCL, pH7.5, and 1 mg/mL BSA) (Invitrogen, CA, USA) to induce DNA strand breaks prior to labeling (following the vendors protocol). Negative control for apoptosis is the nontreated, healthy cells. Isogenic enzyme control is the fixed and permeabilized cells with the labeling solution but without the terminal transferase enzyme.
The Hexane fraction of AG (HAG) drives apoptosis (TUNEL+) of lymphoblasts marginally better in p53+/+ cells compared with p53−/− cells. The TK6 human lymphoblastoid line, NH32 isogenic p53 knockout cells, and Jurkat T cells (dysfunctional p53 pathway) were cultured in RPMI-1640 + 10% NBCS. Cells were exposed to HAG (in triplicate, 𝑛 = 3 per treatment) in RPMI-1640 + 0.1% NBCS for 24 hr at indicated doses and harvested for TUNEL apoptotic assay. The percentage of the cells (Mean ± S.E.) staining positive for TUNEL, indicating apoptosis are given ( 𝑛 = 3 ): (a) TK6 cells; (b) NH32 cells; (c) Jurkat T cells. A minimum of 20,000 events was counted by flow cytometry from each treatment. (d) Confirmation of p53 protein status of the TK6, NH32, and Jurkat T cells. Cell lysates were analyzed by western blot analysis. C+ indicates the positive control for p53, which is an archived HCT-116 cell lysate with wild-type p53. C−, indicates the negative control for p53, which is an archived HCT-116 p53−/− cell lysate with p53 knockout. Results indicate HAG induces apoptosis in lymphoblasts through a limited p53 activity. Significant differences are indicated, where * 𝑃 -value <0.05 and ** 𝑃 -value <0.005, when compared to the control (0 μg/mL) treatment.
2.10. Statistical Analysis
Statistical analysis was done using one-way ANOVA with Scheffe's post hoc test for comparison of endpoint data between mouse groups, as well as in vitro endpoints. The results were analyzed using the Stat-view II statistical program (Abacus Concepts, Inc., Piscataway, NJ, USA) and Microsoft Excel (Microsoft, Bellevue, WA, USA) for Macintosh computers. The 𝑃 value chosen for significance in this study was 0.05.
In UC, intestinal immune responses are often characterized by activation of lamina propria T lymphocytes (LPL) with potent effector functions [43]. Among its regulatory mechanisms, down-regulation of activated T lymphocytes via apoptosis is a very potent and effective strategy, now considered as a key controlling mechanism of UC [5]. To this end, lymphoblastoid cell lines TK6 (WT p53), NH32 (isogenic to TK6, but p53−/−), and Jurkat T cells (which have a dysfunctional p53 pathway) were tested for the apoptosis inducing property of HAG. Previously, we showed the whole AG extract induces apoptosis of TK6, but not NH32 cells [4]. Interestingly, here, we show HAG was able to induce apoptosis (TUNEL+) not only in TK6 cells but (albeit to a lesser extent) also in NH32 cells (Figures 2(a) and 2(b)). HAG induced a 3.9-fold increase in apoptosis of TK6 cells at 500 μg/mL of HAG when compared to the untreated cells. The isogenic p53−/− NH32 cells underwent apoptosis upto 3-fold at 500 μg/mL of HAG when compared to the non-treated cells (Figures 2(a) and 2(b)). Also, at 100 μg/mL, the HAG induced 2.6-fold increase in apoptosis in the TK6 cells, but a 2-fold increase in apoptosis in the NH32 cells. Interestingly, the Jurkat T-cell line (which has a defective p53 pathway) was somewhat resistant to HAG-induced apoptosis (Figure 2(c)), which is consistent with what we have previously observed with the whole AG extract [4]. Figure 2(d) provides confirmation of the p53 status of each of the cell lines used in these experiments.
Overly aggressive CD4+/CD25− T cells are thought to contribute to colitis, and defects in mucosal T-cell apoptosis are likely to be critical in the pathogenesis of colitis [4, 6, 7]. Recently we have shown HAG induces apoptosis of CD4+/CD25− T cells derived from C57/BL6 WT mice during the suppression of colitis [28]. In the present paper, consistent with results from TK6 (p53+/+) and NH32 (p53−/−) cells, HAG induced apoptosis of the CD4+/CD25− T cells isolated from both WT and p53−/− C57/BL6 mice (Table 1). However, the induction of apoptosis was suppressed in the absence of p53. There was 2.6-fold increase From ( 5 . 1 ± 0 . 9 % ) to ( 1 3 . 2 ± 2 . 8 % ) in apoptosis (mean ± S.E.) of effector T cells isolated from p53−/− C57/BL6 mice and a 4.6-fold increase From ( 4 . 5 ± 0 . 5 % ) to ( 2 0 . 7 ± 8 . 9 % ) in the apoptosis was observed in CD4+/CD25− T cells isolated from WT C57/BL6 mice upon treatment with increasing concentration of HAG (0–300 μg/mL).
Dose: hexane fraction of AG (24 hr) Percentage of early apoptotic cells
C57/BL6 p 5 3 − / − Mice C57/BL6 p 5 3 + / + Mice
0 𝜇 g/mL 4 . 1 % 4 . 9 %
100 𝜇 g/mL 5 . 9 % 9 . 8 %
200 𝜇 g/mL 9 . 1 % 1 5 . 7 %
300 𝜇 g/mL 1 8 . 6 % 3 8 . 5 %
Apoptosis (Annexin V positive; propidium iodide negative) of CD4+/CD25− effector T cells from the spleen of p53−/− mice and p53+/+ mice by the increasing concentration of Hexane fraction of AG for 24 hrs.
We have recently shown that HAG suppresses chemically (DSS mouse model) induced colitis in the WT p53 C57/BL6 mice [28]. Previously we have also shown that AG whole extract suppresses chemically induced colitis only in the WT p53 C57/BL6 mice and not in the p53−/− C57/BL6 mice [4]. This led us to the conclusion that the suppression of colitis by AG extract is p53 dependent. We tested the same hypothesis for the HAG. As shown in the Figure 3, the HAG was able to suppress the DSS-induced colitis in both the p53−/− and WT p53 mice. However, consistent with our results on apoptosis of inflammatory cells in vitro, the suppression was more prominent in WT mice than in p53−/− mice. The H&E stained swissrolled colon inflammation score of WT C57/BL6 mice was reduced from 1 5 ± 0 . 8 6 to 4 . 4 ± 0 . 3 2 (average ± S.E.) (a reduction of 71%) in the HAG-treated mice (Figure 3). Similar results were obtained with the p53−/− C57/BL6 mice, where the score dropped from 1 4 . 5 ± 2 . 1 2 to 7 . 8 ± 1 . 7 8 (a reduction of 46%) in the HAG-treated mice. This result is consistent with the notion that the HAG seems to act not only through a p53 pathway but also through a p53-independent pathway while suppressing colitis.
Effect of the HAG on the colon histology score of the acute DSS colitis model. Histological Inflammation score was determined from the H&E stained colon of each group of mice treated with HAG after 2.5 cycles of DSS. Values represent mean ± S.E. Representative H&E stained colon (100x magnified) is shown for each group. Arrows point to areas of inflammation and ulceration. Significant differences are indicated.
Recently we have shown that HAG reduces colon cancer associated with colitis in mice [28]. The mitogenic stimuli triggered signal transduction pathways eventually converge on the cell cycle checkpoint that controls the G1 to S phase transition and activate appropriate cyclin-dependent kinases [19]. This anomaly increases proliferation of the mutated cells to increase the cancer growth and progression. Thus, one way to cause cancer to subside is to prevent this abnormal cancer-cell growth by inducing cell cycle arrest. We tested this hypothesis in vitro by using two different cell lines, HCT-116 p53 WT and HCT-116 p53−/−, and treated these cells with increasing concentrations of HAG for 24 hr. In three repeated experiments, HAG was able to increase a G1 cell cycle arrest in both the WT and p53−/− HCT-116 cells. Representative results are given in Figure 4(a). HAG increased the G1 cell cycle arrest of HCT-116 p53 WT cells 1.7-fold (34% to 56.7%) at 100 μg/mL, HAG treatment and the percentage of cells in G1 remained above 50% with the other concentrations (up to 500 μg/mL) (Figure 4(a)). Similarly there was a 1.5-fold increase (34.1% to 51.2%) of HCT-116 p53−/− cells in G1 phase after treatment with 100 μg/mL of HAG (Figure 4(a)), and this checkpoint remaining with the remaining concentrations. Figure 4(b) provides confirmation of the p53 status of HCT-116 cells used in this experiment.
The HAG induces a G1 checkpoint in colon cancer cells in both p53+/+ and p53−/− colon cancer cells. (a) HCT-116 WT and p53−/− cells were exposed to HAG in RPMI-1640 + 1% NBCS for 24 hr at indicated doses and harvested for cell cycle assay with DAPI stain. Representative percentage of cells in G1, S, and G2/M-phases are indicated for each treatment. A minimum of 20,000 events was counted by flow cytometry from each treatment. (b) Confirmation of p53 protein status of the HCT-116 cells. Cell lysates were analyzed by western blot analysis. C+ indicates the positive control for p53, which is an archived HCT-116 cell lysate with wild-type p53. C− indicates the negative control for p53, which is an archived HCT-116 p53−/− cell lysate with p53 knockout.
Our HAG is extracted from AG whole extract by non-polar solvent 𝑛 -Hexane. Mostly polyacetylenes (panaxynol, panaxydol, and panaxydiol) and fatty acids (FA) present in the AG whole extract were extracted in this fraction [28]. Since HAG is more effective at suppressing colitis than the whole AG extract [28] and based on our previous study, where AG whole extract suppressed colitis through p53-mediated apoptosis of inflammatory cells [4], we hypothesized that HAG also induces apoptosis of inflammatory cells through a p53-mediated mechanism. We pursued our study with HAG and its apoptotic properties with different inflammatory cells. Interestingly, we found that, unlike the whole AG extract, HAG was able to induce modest apoptosis of p53−/− CD4+/CD25− effector T cells and p53−/− lymphoblastoid cells in addition to their p53 WT counterpart cells (Figure 2, Table 1). This indicates that HAG mechanistically acts differently than the AG whole extract, where it shows a modest involvement of p53-dependent apoptosis. This still raises the question, what is the most active component present in the HAG? Recently we have identified the different components present in the HAG, with FAs comprising 43% w/w, polyacetylenes 26.52% w/w (7.39% Panaxydiol, 8.92% Panaxydol, and 10.21% Panaxynol), and less than 0.1% w/w ginsenosides [28]. This indicates that specific FA ingredients or specific polyacetylenes, or both are responsible for the apoptotic property of HAG through both p53-dependent and p53-independent mechanisms.
Interestingly, Wong et al. have shown that Asian ginseng extracted with ethanol induces a G2-M arrest and apoptosis via modulation of MAPK and p53 pathway in LLC-1 cells (Mouse Lewis lung carcinoma cells) [44]. The AG whole extract is also extracted with ethanol and the ginsenoside composition is for the most part, similar in Asian and American ginseng. Separate studies from Kim et al. [45, 46] have shown that the ginsenosides-Rs3 and -Rs4 selectively elevate protein levels of p53 and p21WAF1, and downregulate the activities of the cyclin-dependent kinases, resulting in cell cycle arrest at the G1/S boundary and apoptosis of SK-HEP-1 cells (immortalized human hepatoma cells) [45, 46]. All these observations suggest that ginsenosides may be a key player in modulating apoptosis and cell cycle through the p53 pathway and are consistent with the findings here and our previous findings that our whole AG extract has 10.1% ginsenoside (w/w) compared to the HAG which has 0.074% ginsenoside (w/w) content [28].
Interestingly, ginsenoside Rd, which is present in AG whole extract (extracted with aqueous ethanol) and absent in the HAG, has been shown to attenuate the inflammatory response to TNBS− (2,4,6-trinotrobenzenesulfonic acid-) induced colitis by down-regulating multiple pro-inflammatory cytokines through the modulation of JNK (c-Jun N-terminal kinase) and p38 activation [47]. Similarly another ginsenoside not present in HAG, Rb1, and its metabolite compound K after oral administration blocked TNBS-induced expression of iNOS (inducible-nitric oxide synthase), Cox-2 (cyclooxygenase-2), and NF-κβ (nuclear transcription factor κβ) activation in mice [48]. Ginsenoside Rb1 and its metabolite compound inhibited the activation of key inflammatory mediators IRAK-1 (interleukin-1 receptor associated kinase-1), IKK-β (inhibitor of NF-κβ kinase), NF-κβ, and MAPK (mitogen-activated protein kinases) while suppressing colitis [48]. These studies suggest that different ginsenosides may help attenuate inflammation, and our finding that 𝑛 -Hexane does not extract these ginsenosides from ginseng should not rule out that ginsenosides are not the active components of ginseng. Taken together, these observations suggest that multiple components of AG, including ginsenosides, FAs, and polyacetylenes, are responsible for the activity of AG in the suppression of colitis in animals. Further studies are necessary to delineate the most active ingredient(s)/molecule(s).
Moon et al. have shown that during the G1/S cell cycle arrest, protein levels of p21WAF1, p16INK4A, p53, pRb (retinoblastoma protein), and E2F-1 (transcription factor E2F1) were not changed after exposure to the polyacetylene, panaxydol (isolated from P. ginseng), in the human malignant melanoma cell line, SK-MEL-1 [49]. Human promyelocytic leukemia HL60 cells do not express the p53 protein due to a large deletion in the gene [50]. The polyacetylenes, panaxynol, and panaxydol (isolated from the lipophilic fractions of Panax notoginseng) have been shown to inhibit the proliferation of HL60 cells in a time− and dose-dependent manner via an apoptotic pathway [51]. These studies indicate that polyacetylenes may act independently of p53 while inducing apoptosis of certain cell lines. HAG-induced apoptosis indicated by cleaved PARP in TK6 cells, appears to change the protein levels of WT p53 and activated form of p53 (phospho-Ser15) very little [28]. After 24 hr of HAG treatment (300 μg/mL), we observed a 2.6-fold increase in apoptosis of p53−/− CD4+/CD25− T cells and a 4.6-fold increase in apoptosis of p53+/+ CD4+/CD25− T cells when compared to untreated cells (Table 1). Figures 2(a), 2(b) and 2(c), and our recent study [28] are consistent and indicate that p53 has a limited role in inducing HAG-mediated apoptosis of inflammatory cells both in vitro and ex vivo. A clearly increased p53, phospho-p53 Ser-15, and cleaved PARP protein levels along with the increased apoptosis were reported with the AG whole extract, suggesting p53 plays a key role in apoptosis of inflammatory cells induced by the whole AG extract [4]. However, an increased cleaved PARP protein expression and only a slight change in the p53, phospho-p53 (Ser-15), and p53-upregulated mediator of Apoptosis (PUMA) protein expression of TK6 cells after treatment with HAG [28] are consistent with our current results that HAG induces apoptosis but this induction of apoptosis is likely mediated through both p53-dependent and p53-independent mechanisms.
The other high volume components present in HAG are FAs, where 19% w/w is linoleic acid, a polyunsaturated fatty acid [28]. Kwon et al. have reported that linoleic acid treatment resulted in a concentration-dependent growth inhibition of AGS cells (human gastric adenocarcinoma cells) by inducing apoptosis in a p53-independent manner with an elevated Fas and Fas ligand expression [52]. This, then, is consistent with our results regarding a limited role for p53 in the cells undergoing apoptosis after treatment with HAG.
We further tested our compound in the in vivo animal study. As indicated (Figure 1), both the p53−/− and p53 WT C57/BL6 mice were subjected to 2.5 cycles of 1% DSS to induce colitis. In this preventive DSS model, 75 ppm (11.9 mg/kg) of HAG or vehicle (1 × PBS) were administered to the mice daily (PO) and continued throughout the course of the experiment. In our previous published studies, AG whole extract was effective in suppressing colitis only in the p53 WT C57/BL6 mice where whole AG extract prevented colon epithelial cells from the DNA damage due to DSS and induced apoptosis of lymphocytes in vivo [4]. In the present study, with the same protocol, the HAG was able to suppress DSS-induced colonic inflammation to approximately one-third (from 1 5 ± 0 . 8 6 to 4 . 4 ± 0 . 3 2 ). Interestingly, the HAG also showed a modest suppression of inflammation to about one-half (from 1 4 . 5 ± 2 . 1 2 to 7 . 8 ± 1 . 7 8 ) in the p53−/− C57/BL6 mice (Figure 3). This provides evidence that the HAG is anti-inflammatory and p53 plays a limited role in the HAG-mediated suppression of colitis.
Apoptosis is an ordered cellular process that occurs in various physiological and pathological conditions. Two major problems are associated with apoptosis; too much apoptosis is associated with various degenerative diseases and too little to no apoptosis is associated with carcinogenesis [53]. Hence one way to tackle the cancerous cells is to induce apoptosis of such cells. Colon cancer is associated with long standing UC and hence inflammation seems to drive the progression of cancer. Interestingly, both HCT-116 p53+/+ and p53−/− colon cancer cells were relatively resistant to apoptosis induced by HAG (data not shown). However, there was a G1 cell cycle checkpoint with HAG treatment (Figure 4(a)). It is unlikely that the HAG causes DNA damage to cause this checkpoint. It is more likely that one or more ingredients in HAG target modulators of the cell cycle (e.g., cyclins or cyclin-dependent kinases). Experiments are ongoing to examine this possibility. Consistent with our findings, Kang et al. have shown that lipid soluble ginseng extract (red ginseng extracted with 𝑛 -Hexane) induces a cell cycle arrest at the G1 phase in NCI-H460 cells (human lung cancer cells) [54]. These separate observations suggest that the HAG may halt the progression of cell cycle at G1 phase in multiple cancer cell lines.
We have shown here that the HAG can induce apoptosis in lymphoblastoid cells and CD4+/CD25− effector T cells and cause a G1 checkpoint in colon cancer cell lines. It also suppressed chemically induced colitis in C57/BL6 mice. In all the above observations, we simultaneously tested the pro-apoptotic and anti-inflammatory properties of HAG with the WT p53 and its p53−/− counterpart cells and mice. It is evident that HAG can perform its pro-apoptotic, anti-inflammatory, and cell cycle arrest activities even in the absence of p53. However, the HAG has a more robust effect in the presence of p53. These observations suggest that there is a role of p53 in the HAG-mediated apoptosis of inflammatory cells and suppression of colitis; but this role is limited. This entails that the HAG affects other pathways independent of p53 in the suppression of colitis. The reasoning for this can only be speculated. For example, is there one component in the whole AG extract that is particularly powerful in inducing p53-mediated apoptosis that is missing from the HAG? Does this component inhibit the ability of components only seen in the HAG fraction to drive p53-independent apoptosis and suppress p53-independent colitis? This is significant because the HAG suppresses colitis and associated colon cancer [28]. Because mutation in p53 is observed in most cancers, including colitis-driven colon cancer, the HAG may be particularly effective in targeting both p53+/+ and p53−/− inflammatory cells and cancer cells and hence particularly effective in inhibiting the colitis-to-cancer sequence.
None of the author of this paper have a relation with the commercial identities used in this paper.
This work was supported by the Center for CAM Research on Autoimmune and Inflammatory Diseases, NIH Grant 1P01AT003961-01A1 (P. S. Nagarkatti, L. J. Hofseth, M. Nagarkatti), and the COBRE funded University of South Carolina Center for Colon Cancer Research, NIH Grant P20RR17698-01 (Franklin Berger, Director). Thanks are also due to the P20RR17698-01 Statistical Core (Dr. Edsel Pena, Director), and 1P01AT003961-01A1 Immunotoxicology Core (Dr. Narendra Singh, Director), P20RR17698-01 Mouse Core (Dr. Marj Pena, Director), and P20RR17698-01 Imaging/Histology Core.
Supplementary Figure 1 represents the purity of CD4+/CD25− T cells isolated from the spleen of the mice. Supplementary Table 1 represents the Inflammation Scores from individual animals based on the inflammation severity, extent, crypt damage and percent involvement. (Inflammation score range: 0–40, as described in the methods section).
Supplementary Figure
Supplementary Table
C. L. Maynard and C. T. Weaver, "Intestinal effector T cells in health and disease," Immunity, vol. 31, no. 3, pp. 389–400, 2009. View at: Publisher Site | Google Scholar
R. Eri, M. A. McGuckin, and R. Wadley, "T cell transfer model of colitis: a great tool to assess the contribution of T cells in chronic intestinal inflammation," Methods in Molecular Biology, vol. 844, pp. 261–275, 2012. View at: Publisher Site | Google Scholar
R. H. Siggers and D. J. Hackam, "The role of innate immune-stimulated epithelial apoptosis during gastrointestinal inflammatory diseases," Cellular and Molecular Life Sciences, vol. 68, no. 22, pp. 3623–3634, 2011. View at: Publisher Site | Google Scholar
Y. Jin, A. B. Hofseth, X. Cui et al., "American ginseng suppresses colitis through p53-mediated apoptosis of inflammatory cells," Cancer Prevention Research, vol. 3, no. 3, pp. 339–347, 2010. View at: Publisher Site | Google Scholar
A. D. Levine, "Apoptosis: implications for inflammatory bowel disease," Inflammatory Bowel Diseases, vol. 6, no. 3, pp. 191–205, 2000. View at: Google Scholar
R. B. Sartor, "Mechanisms of disease: pathogenesis of Crohn's disease and ulcerative colitis," Nature Clinical Practice Gastroenterology and Hepatology, vol. 3, no. 7, pp. 390–407, 2006. View at: Publisher Site | Google Scholar
M. G. Neuman, "Immune dysfunction in inflammatory bowel disease," Translational Research, vol. 149, no. 4, pp. 173–186, 2007. View at: Publisher Site | Google Scholar
A. H. Wyllie, J. F. R. Kerr, and A. R. Currie, "Cell death: the significance of apoptosis," International Review of Cytology, vol. 68, pp. 251–306, 1980. View at: Google Scholar
D. Brenner and T. W. Mak, "Mitochondrial cell death effectors," Current Opinion in Cell Biology, vol. 21, no. 6, pp. 871–877, 2009. View at: Publisher Site | Google Scholar
N. Özören and W. S. El-Deiry, "Cell surface death receptor signaling in normal and cancer cells," Seminars in Cancer Biology, vol. 13, no. 2, pp. 135–147, 2003. View at: Publisher Site | Google Scholar
R. Dirisina, R. B. Katzman, T. Goretsky, E. Managlia, N. Mittal, D. B. Williams et al., "p53 and PUMA independently regulate apoptosis of intestinal epithelial cells in patients and mice with colitis," Gastroenterology, vol. 141, no. 3, pp. 1036–1045, 2011. View at: Publisher Site | Google Scholar
L. J. Hofseth, S. Saito, S. P. Hussain et al., "Nitric oxide-induced cellular stress and p53 activation in chronic inflammation," Proceedings of the National Academy of Sciences of the United States of America, vol. 100, no. 1, pp. 143–148, 2003. View at: Publisher Site | Google Scholar
S. P. Hussain, L. J. Hofseth, and C. C. Harris, "Radical causes of cancer," Nature Reviews Cancer, vol. 3, no. 4, pp. 276–285, 2003. View at: Google Scholar
B. A. Lashner, W. M. Bauer, L. A. Rybicki, and J. R. Goldblum, "Abnormal p53 immunohistochemistry is associated with an increased colorectal cancer-related mortality in patients with ulcerative colitis," The American Journal of Gastroenterology, vol. 98, no. 6, pp. 1423–1427, 2003. View at: Publisher Site | Google Scholar
C. Alkim, B. Savas, A. Ensari et al., "Expression of p53, VEGF, microvessel density, and cyclin-D1 in noncancerous tissue of inflammatory bowel disease," Digestive Diseases and Sciences, vol. 54, no. 9, pp. 1979–1984, 2009. View at: Publisher Site | Google Scholar
Y. Jin, V. S. Kotakadi, L. Ying et al., "American ginseng suppresses inflammation and DNA damage associated with mouse colitis," Carcinogenesis, vol. 29, no. 12, pp. 2351–2359, 2008. View at: Publisher Site | Google Scholar
V. S. Kotakadi, Y. Jin, A. B. Hofseth et al., "Ginkgo biloba extract EGb 761 has anti-inflammatory properties and ameliorates colitis in mice by driving effector T cell apoptosis," Carcinogenesis, vol. 29, no. 9, pp. 1799–1806, 2008. View at: Publisher Site | Google Scholar
F. Bunz, P. M. Hwang, C. Torrance et al., "Disruption of p53 in human cancer cells alters the responses to therapeutic agents," The Journal of Clinical Investigation, vol. 104, no. 3, pp. 263–269, 1999. View at: Google Scholar
M. Andreeff, D. W. Goodrich, and A. B. Pardee, "Cell proliferation, differentiation, and apoptosis," in Holland-Frei Cancer Medicine, R. C. Bast, D. W. Kufe, P. E. Pollock, R. R. Weicheelbaum, J. F. Holland, E. Frei et al., Eds., BC Decker, Ontario, Canada, 5th edition, 2000. View at: Google Scholar
M. B. Kastan, O. Onyekwere, D. Sidransky, B. Vogelstein, and R. W. Craig, "Participation of p53 protein in the cellular response to DNA damage," Cancer Research, vol. 51, no. 23, part 1, pp. 6304–6311, 1991. View at: Google Scholar
S. J. Kuerbitz, B. S. Plunkett, W. V. Walsh, and M. B. Kastan, "Wild-type p53 is a cell cycle checkpoint determinant following irradiation," Proceedings of the National Academy of Sciences of the United States of America, vol. 89, no. 16, pp. 7491–7495, 1992. View at: Google Scholar
A. R. Clarke, C. A. Purdie, D. J. Harrison et al., "Thymocyte apoptosis induced by p53-dependent and independent pathways," Nature, vol. 362, no. 6423, pp. 849–852, 1993. View at: Publisher Site | Google Scholar
S. W. Lowe, E. M. Schmitt, S. W. Smith, B. A. Osborne, and T. Jacks, "p53 is required for radiation-induced apoptosis in mouse thymocytes," Nature, vol. 362, no. 6423, pp. 847–849, 1993. View at: Publisher Site | Google Scholar
M. S. Greenblatt, W. P. Bennett, M. Hollstein, and C. C. Harris, "Mutations in the p53 tumor suppressor gene: clues to cancer etiology and molecular pathogenesis," Cancer Research, vol. 54, no. 18, pp. 4855–4878, 1994. View at: Google Scholar
V. A. Assinewe, B. R. Baum, D. Gagnon, and J. T. Arnason, "Phytochemistry of wild populations of Panax quinquefolius L. (North American ginseng)," Journal of Agricultural and Food Chemistry, vol. 51, no. 16, pp. 4549–4553, 2003. View at: Publisher Site | Google Scholar
W. E. Court, "Ginseng: the history of an insignificant plant," Pharmaceutical Historian, vol. 30, no. 3, pp. 38–44, 2000. View at: Google Scholar
B. Li, C. Z. Wang, T. C. He, C. S. Yuan, and W. Du, "Antioxidants potentiate American ginseng-induced killing of colorectal cancer cells," Cancer Letters, vol. 289, no. 1, pp. 62–70, 2010. View at: Publisher Site | Google Scholar
D. Poudyal, P. Mai Le, T. Davis, A. B. Hofseth, A. Chumanevich, A. A. Chumanevich et al., "A Hexane fraction of American ginseng suppresses mouse colitis and associated colon cancer: anti-inflammatory and proapoptotic mechanisms," Cancer Prevention Research, vol. 5, no. 4, pp. 685–696, 2012. View at: Publisher Site | Google Scholar
S. P. Hussain and C. C. Harris, "p53 biological network: at the crossroads of the cellular-stress response pathway and molecular carcinogenesis," Journal of Nippon Medical School, vol. 73, no. 2, pp. 54–64, 2006. View at: Publisher Site | Google Scholar
A. J. Schetter, N. H. Heegaard, and C. C. Harris, "Inflammation and cancer: interweaving microRNA, free radical, cytokine and p53 pathways," Carcinogenesis, vol. 31, no. 1, Article ID bgp272, pp. 37–49, 2010. View at: Publisher Site | Google Scholar
T. R. Skopek, H. L. Liber, B. W. Penman, and W. G. Thilly, "Isolation of a human lymphoblastoid line heterozygous at the thymidine kinase locus: possibility for a rapid human cell mutation assay," Biochemical and Biophysical Research Communications, vol. 84, no. 2, pp. 411–416, 1978. View at: Google Scholar
Y. Y. E. Chuang, Q. Chen, and H. L. Liber, "Radiation-induced mutations at the autosomal thymidine kinase locus are not elevated in p53-null cells," Cancer Research, vol. 59, no. 13, pp. 3073–3076, 1999. View at: Google Scholar
U. Schneider, H. U. Schwenk, and G. Bornkamm, "Characterization of EBV-genome negative "null" and "T" cell lines derived from children with acute lymphoblastic leukemia and leukemic transformed non-Hodgkin lymphoma," International Journal of Cancer, vol. 19, no. 5, pp. 621–626, 1977. View at: Google Scholar
J. Cheng and M. Haas, "Frequent mutations in the p53 tumor suppressor gene in human leukemia T-cell lines," Molecular and Cellular Biology, vol. 10, no. 10, pp. 5502–5509, 1990. View at: Google Scholar
R. Laumann, M. Jucker, and H. Tesch, "Point mutations in the conserved regions of the p53 tumour suppressor gene do not account for the transforming process in the Jurkat acute lymphoblastic leukemia T-Cells," Leukemia, vol. 6, no. 3, pp. 227–228, 1992. View at: Google Scholar
X. Cui, Y. Jin, A. B. Hofseth et al., "Resveratrol suppresses colitis and colon cancer associated with colitis," Cancer Prevention Research, vol. 3, no. 4, pp. 549–559, 2010. View at: Publisher Site | Google Scholar
S. Reagan-Shaw, M. Nihal, and N. Ahmad, "Dose translation from animal to human studies revisited," The FASEB Journal, vol. 22, no. 3, pp. 659–661, 2008. View at: Publisher Site | Google Scholar
G. N. Predy, V. Goel, R. Lovlin, A. Donner, L. Stitt, and T. K. Basu, "Efficacy of an extract of North American ginseng containing poly-furanosyl-pyranosyl-saccharides for preventing upper respiratory tract infections: a randomized controlled trial," Canadian Medical Association Journal, vol. 173, no. 9, pp. 1043–1048, 2005. View at: Publisher Site | Google Scholar
V. Vuksan, M. P. Stavro, J. L. Sievenpiper et al., "American ginseng improves glycemia in individuals with normal glucose tolerance: effect of dose and time escalation," Journal of the American College of Nutrition, vol. 19, no. 6, pp. 738–744, 2000. View at: Google Scholar
L. A. Dieleman, M. J. Palmen, H. Akol et al., "Chronic experimental colitis induced by dextran sulphate sodium (DSS) is characterized by Th1 and Th2 cytokines," Clinical and Experimental Immunology, vol. 114, no. 3, pp. 385–391, 1998. View at: Publisher Site | Google Scholar
O. Morteau, S. G. Morham, R. Sellon et al., "Impaired mucosal defense to acute colonic injury in mice lacking cyclooxygenase-1 or cyclooxygenase-2," The Journal of Clinical Investigation, vol. 105, no. 4, pp. 469–478, 2000. View at: Google Scholar
L. Ying, J. Marino, S. P. Hussain et al., "Chronic inflammation promotes retinoblastoma protein hyperphosphorylation and E2F1 activation," Cancer Research, vol. 65, no. 20, pp. 9132–9136, 2005. View at: Publisher Site | Google Scholar
C. Fiocchi, "Inflammatory bowel disease: etiology and pathogenesis," Gastroenterology, vol. 115, no. 1, pp. 182–205, 1998. View at: Publisher Site | Google Scholar
V. K. Wong, S. S. Cheung, T. Li et al., "Asian ginseng extract inhibits in vitro and in vivo growth of mouse lewis lung carcinoma via modulation of ERK-p53 and NF-κB signaling," Journal of Cellular Biochemistry, vol. 111, no. 4, pp. 899–910, 2010. View at: Publisher Site | Google Scholar
S. E. Kim, Y. H. Lee, J. H. Park, and S. K. Lee, "Ginsenoside-Rs4, a new type of ginseng saponin concurrently induces apoptosis and selectively elevates protein levels of p53 and p21(WAF1) in human hepatoma SK-HEP-1 cells," European Journal of Cancer, vol. 35, no. 3, pp. 507–511, 1999. View at: Publisher Site | Google Scholar
S. E. Kim, Y. H. Lee, J. H. Park, and S. K. Lee, "Ginsenoside-Rs3, a new diol-type ginseng saponin, selectively elevates protein levels of p53 and p21WAF1 leading to induction of apoptosis in SK-HEP-1 cells," Anticancer Research, vol. 19, no. 1, pp. 487–491, 1999. View at: Google Scholar
X. L. Yang, T. K. Guo, Y. H. Wang, Y. H. Huang, X. Liu, X. X. Wang et al., "Ginsenoside Rd attenuates the inflammatory response via modulating p38 and JNK signaling pathways in rats with TNBS-induced relapsing colitis," International Immunopharmacology, vol. 12, no. 2, pp. 408–414, 2012. View at: Publisher Site | Google Scholar
E. H. Joh, I. A. Lee, I. H. Jung, and D. H. Kim, "Ginsenoside Rb1 and its metabolite compound K inhibit IRAK-1 activation—the key step of inflammation," Biochemical Pharmacology, vol. 82, no. 3, pp. 278–286, 2011. View at: Publisher Site | Google Scholar
J. Moon, S. J. Yu, H. S. Kim, and J. Sohn, "Induction of G1 cell cycle arrest and p2KIP1 increase by panaxydol isolated from Panax ginseng," Biochemical Pharmacology, vol. 59, no. 9, pp. 1109–1116, 2000. View at: Publisher Site | Google Scholar
D. Wolf and V. Rotter, "Major deletions in the gene encoding the p53 tumor antigen cause lack of p53 expression in HL-60 cells," Proceedings of the National Academy of Sciences of the United States of America, vol. 82, no. 3, pp. 790–794, 1985. View at: Google Scholar
Z. Yan, R. Yang, Y. Jiang et al., "Induction of apoptosis in human promyelocytic leukemia HL60 cells by panaxynol and panaxydol," Molecules, vol. 16, no. 7, pp. 5561–5573, 2011. View at: Publisher Site | Google Scholar
J. I. Kwon, G. Y. Kim, K. Y. Park, C. H. Ryu, and Y. H. Choi, "Induction of apoptosis by linoleic acid is associated with the modulation of Bcl-2 family and Fas/FasL system and activation of caspases in AGS human gastric adenocarcinoma cells," Journal of Medicinal Food, vol. 11, no. 1, pp. 1–8, 2008. View at: Publisher Site | Google Scholar
R. S. Wong, "Apoptosis in cancer: from pathogenesis to treatment," Journal of Experimental & Clinical Cancer Research, vol. 30, p. 87, 2011. View at: Publisher Site | Google Scholar
M. R. Kang, H. M. Kim, J. S. Kang et al., "Lipid-soluble ginseng extract induces apoptosis and G0/G1 cell cycle arrest in NCI-H460 human lung cancer cells," Plant Foods for Human Nutrition, vol. 66, no. 2, pp. 101–106, 2011. View at: Publisher Site | Google Scholar
Copyright © 2012 Deepak Poudyal et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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Mugabe government to seize Zimbabwe's diamond operations
Just days after celebrating his 92nd birthday, Zimbabwe's President Robert Mugabe says his successor must be chosen democratically and that his wife will not automatically inherit his role.
"Companies that have been mining diamonds have robbed us of our wealth. That is why we have now said the state must have a monopoly."
In a televised interview Mugabe said his government would take possession of all diamond operations because existing miners had robbed the country of its wealth.
"In the mining sector, the mining of diamonds will be reserved for the state, be a matter for the state alone," Mugabe told state broadcaster ZBC TV. "We have not received much from the diamond industry not much by way of earnings. I don't think we have received two billion dollars."
Mugabe's comments come one week after the country's ministry of mines ordered all mining companies to halt work and leave the Marange diamond fields, saying their licences had not been renewed. The ministry denied it was seizing the mines.
Mugabe said the Zimbabwean state will now own all the diamonds in the country.
Mnangagwa urges Zimbabwe to unite behind his presidency
Zimbabwe election challenge
27 Zimbabwe opposition members charged


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Published On: Mon, Jun 24th, 2013
Business / US News | By Brandon Jones
Twinkies confirmed to be returning to the shelf in July
Hostess may be betting on a sweet comeback for Twinkies when they return to shelves next month.
The company that went bankrupt after a failed battle with unionized workers last year is back with new owners and a lean business model.
Twinkies photo by Mamiejeanjean
Based on the outpouring of nostalgia sparked by its demise, Hostess is expecting a blockbuster return next month for Twinkies and other sugary treats, such as CupCakes and Donettes. The company says the cakes will taste the same but that the boxes will now bear the tag line "The Sweetest Comeback In The History Of Ever."
"A lot of impostor products have come to the market while Hostess has been off the shelves," says Daren Metropoulos, a principal of the investment firm Metropoulos & Co., which teamed up with Apollo Global Management to buy a variety of Hostess snacks.
Twinkies and other snack cakes back on shelves starting July 15.
Hostess Brands Inc. was struggling for years before it filed for Chapter 11 bankruptcy reorganization in early 2012.
Workers blamed the troubles on years of mismanagement, as well as a failure of executives to invest in brands to keep up with changing tastes. The company said it was weighed down by higher pension and medical costs than its competitors, whose employees weren't unionized.
Hostess hired restructuring expert Greg Rayburn as its CEO. But Rayburn ultimately failed to reach a contract agreement with its second largest union. In November, he blamed striking workers for crippling the company's ability to maintain normal production and announced that Hostess would liquidate.
The announcement triggered a rush on Hostess snack cakes, with stores selling out of the most popular brands within hours.
About 15,000 unionized workers lost their jobs in the aftermath.
In unwinding its business, Hostess sold off its brands in chunks to different buyers.
Its major bread brands including Wonder were sold to Flowers Foods, which makes Tastykakes. McKee Foods, which makes Little Debbie snack cakes, snapped up Drake's Cake, which includes Devil Dogs and Yodels.
Metropoulos & Co. and Apollo bought Twinkies and other Hostess cakes for $410 million.
Business NewsHostess bankruptHostess closingHostess strikeTwinkiesunion
Election 2020: Cory Booker exits race, Steve Phillips blames 'White man's bump,' racism
Immigration bill includes amendment to give away free cars, bicycles, scooters … | WordPress.org A semantic personal publishing platform with a focus on aesthetics, web standards, and usability. says:


Everything those teammates did was for the glory of Lance. Millers targets will be a strong group of receivers including speedy junior Ben Douglas, who is making the jump from junior varsity, and 6-foot-4-inch Mike Clenshaw. nike air max tn black orange To celebrate the anniversary, the company is honoring long-term employees at a breakfast. The Duals were a great alternative, but after a few months, I decided I wanted something for more di.


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Stuart Durkheimer (1921-2009)
Stuart Durkheimer was the second son of Sylvan and Dorothy Durkheimer. His paternal grandfather had come from Bavaria in the mid-19th century and was one of the early Jewish pioneers in Oregon. Stuart and his siblings, James, Marian (Jaffe) and Eloise (Spiegel) grew up in Northwest Portland and attended Chapman Elementary and Lincoln High School. The family spent summers at Seaview, Washington and they were lifelong members of Congregation Beth Israel.
He graduated from Stanford University in 1942, and being unable to serve in the military during the Second World War because of poor eyesight, he instead began his career in the grocery business, eventually becoming president and CEO of Wadhams & Company. Following the sale of Wadhams, Stuart served as president/CEO of Coast and General Distributors.
The Boy Scouts greatly influenced Stuart's life and as an Eagle Scout he traveled to Europe for the World Jamboree in 1937. As a young adult he became the leader of troop 69. He also served on the boards of numerous organizations in the community including The Oregon Symphony, Oregon Jewish Community Foundation, Robison Board, Friendly-Rosenthal Foundation, Jewish Federation, and was president of Temple Beth Israel and founding president of the Oregon Jewish Committee. He was a member of the Multnomah Athletic Club, Tualatin Country Club, Astoria Country Club and Congregation Beth Israel. He was also a Little League baseball coach at Wallace Park.
Stuart married Leah Kinspel in 1943, and they had Barbara (m. Gary Larsen), Alan, and Julie (m. Mark Dohrmann). They had a second home at Arch Cape, Oregon, where they spent summers with their children entertaining. After Leah's death in 2000, Stuart had a long relationship with Rosemarie Rosenfeld.


This web display every caledarios annual, including 2018 - 2020. This shall useful if thee are looking for a specific date (When there is a vacation / checking routine health care schedule) or probably you want to know what the week number of a date in 2018 is.You might also use this web to look out when a distinctive day or date in 2018 takes place.
Printable HD Kenya Flag have some pictures that related each other in the collection of images below. Find out the most recent image for print Kenya Flag, and also you can have the picture here which we summarize from various other sites. http://a.lireepub.website just summarizes and does not save on our database.


The lovely Sarah tagged us to reveal what we carry around on our handbags in this post and I jumped at the opportunity! I really love seeing what is in other people's handbags and comparing it to the junk that I carry around in mine. Recently I have been rotating 3 bags, dependent on the outfit that I am wearing and also what I need to lug to uni.
This red Topshop bag which I swapped with my sister I wear whenever I can as it's the perfect size for everyday.
This tan Asos bag is absolutely HUGE! I can fit all my folders for uni inside it which is quite an achievement!
As you can tell, I have a LOT of stuff in my bag! This is the contents from Friday arranged neatly so I can photograph them for you all!
Some of my make-up. The Rimmel mascara is my favourite daytime mascara at the moment and the Topshop lipstick is in 'Blush'. I have used an eyebrow pencil since a beautician got a little bit overzealous waxing my eyebrows and I had to fill them in! I thoroughly recommend it though, it just makes me look more polished, even when I'm not wearing make-up.
What do you have in your bag? Do you carry everything but the kitchen sink, or just the bare essentials? I tag anyone who wants to share what's in their bag!
I love thiese types of posts: it's so interesting to see what's in peoples bags. The tanned ASOS bag is gorgeous. I'm taking ideas from this: I think I should keep hair producrs in my bag as well.


Our Team- Metro House Builders.com ~ Dream It, Design It, Build It!
Joe graduated from Weymouth Vocational High School in 1982 with a diploma in metal technology and worked full time as welder and fabricator for 15 years. In 1997 Joe began working part time, diversifying into all fields of construction, from laboring to carpentry with Scott Condo, a local builder. Scott immediately recognized Joe's talent and offered him full time placement in 1998 and they enjoyed working together until 2000 when Scott passed away unexpectedly. Paul Wilkes and Scott's team had collaborated on many successful condominium projects since the mid 90's and Paul welcomed Joe into the Metro team shortly thereafter. Paul recognized Joe as being production oriented, focused and logically minded. Joe graduated in 2004 from the Construction Certification Institute, Inc. adding to his list of accomplishments. Described as a passionate early-riser by his fellow builders, Joe's talent and intellect earned him a Construction Supervisor License from the Commonwealth of Massachusetts in 2005.
Starting with a prominent beauty supply and consultation company in 1999, Ken grew from a customer service rep to Director of Marketing in just a few years. In 2003 he helped CEO Elda Argenti launch Plantogen, a plant derived skincare company from New Zealand before joining another skincare company in 2009 helping CEO Mitch Bloom transform his tanning solutions brand into a platform of education and business change. A teacher, a cheerleader, and a designer with a keen mind and talent in almost ever sector of business, Ken has worked closely with Metro founder, Paul Wilkes since 2006 in transforming Metro's marketing approach and launching Metro's virtual presence for the first time.
I was introduced to Metropolitan Building Systems Corporation through a client who was completing an extensive addition and renovation of a suburban home. I was immediately impressed with Paul Wilkes' building knowledge. I have subsequently recommended Paul to clients who were completing renovations in a variety of building types with varying project scopes. Paul approaches each project with careful consideration, presents options, and works collaboratively with the home owner, design professionals and the trades.
As someone who has used Metropolitan Building Systems services for over 10 years, I can assure you that Metropolitan Building Systems attention to detail and problem resolution skills are outstanding and much appreciated. Always sensitive to deadlines and adapting to any project changes, portrays extreme competence and professionalism at all times. Not only are they knowledgeable in this technical field, they are also an excellent communicator and attentive to detail, always going the extra mile to ensure customer service that is beyond expectations.
Owner / I.S. Hernandez Design Services, Inc.
...thank you again for the great job you did on our house...It was also nice to have three of the same workers on the job from the start to finish, and the time spent on the little things...that shows a great diversity in the crews skill. To sum it up, we'd never think that we could come in contact with such a professional...company that left us feeling so secure about our home.
...Thanks again for a superb job - i'm absolutely thrilled by the way the house looks...Thanks again for everything.
...Metro Building Systems Corp. reconstructed my home 5 years ago....I found his work to be of a high quality. (In fact, my architect commented, following her initial walk through that the framing work was excellent).
I am an Ipswich based architect, specializing in residential projects. I have always found Paul to be a capable and competitive builder with excellent communication skills and high standards for workmanship and detail. I would not hesitate to recommend them for any construction project.
As an experienced construction project manager...I feel I have a qualified background to provide this accurate testimonial...Metro excels in investigation, pricing, quality assurance, construction management...I am confident that any homeowner or other property manager will also realize the same...satisfaction once they retain Metro.
Vice President / Vintage Properties, Inc.
From the very beginning, Paul set a tone of professionalism and personal warmth....There was never an issue with communication; in fact we felt like we were part of a team...Paul consistently went "the extra mile" with his time, patience, availability, and support.
Copyright ©2015 - Metropolitan Building Systems, Corp.


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HERITAGE - LIBRARY MANAGEMENT SOFTWARE
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Our Proud Users!
Originally founded by Danish engineer Ove Arup in 1946, Arup is now one of the leading engineering organisations in the world, employing more than 6,500 people internationally with an annual turnover exceeding £280 million. Better known for involvement with many of the high profile buildings of the post-war years, Arup has made a name for itself in many other aspects of engineering from roads and bridges to indestructible flasks for transporting nuclear waste.
Arup have a number of offices around the UK and associated company around the globe, with a main library located at the London office. This library holds over 150,000 items, including books, pamphlets, CDs and serials.
Julian Dawson, the librarian at Arup's London office, began his search for a replacement library system in January 2002, when he visited the British Library Exhibition for Small and Special Libraries in London. He had seen Heritage previously at the Library Resources Exhibition in 1997, but it was not until the decision was made to replace DB Textworks, Arup's existing system, that he investigated further.
Julian identified three management systems that would potentially suit the requirements of his library and invited the suppliers to give on-site demonstrations to both himself and his library staff. He had a number of criterion upon which he assessed each system, including "reliability, ease of use, good functionality, value for money [and] good support" and had prioritised "robustness and level of support" as the most important factors.
Julian was impressed with both the sophistication and price of Heritage, and after reviewing all the short listed systems he invited IS Oxford back to discuss their specific requirements in more detail. This meeting enabled each of the library staff to satisfy themselves that Heritage would meet their individual requirements. IS Oxford then provided an evaluation copy of the software for the library staff to trial. Confident that the testing had gone well, Julian decided to go ahead with Heritage.
"[Heritage] did most of what we wanted far more cheaply than other systems. We liked the approach of the company, and its ethos; and its commitment to the customer."
Heritage was installed at Arup in April 2002.
"[The] installation process was very straightforward. The Server Performance Pack service proved more of a problem, but support was able to spend time sorting this out. Configurations of some PCs had to be changed, but not a serious issue."
Julian and his staff had a number of training sessions, which they found to be "generally very good" and they have arranged more for the future. They have also made use of the support line, which they initially found to be too slow "when our system is down and therefore support requirement is urgent". This issue was addressed with IS support staff, and together with Julian they have "established ways to improve things from both sides". IS staff are proud of their support reputation and are very keen to offer the best service possible, so a variety of methods of support are available (e.g. via telephone, email, the Internet etc) to cater for individual needs. They continually monitor the effectiveness of our support service and welcome customer feedback as part of this process.
Having used Heritage for a number of years now, we recently contacted Julian to ask him how he now feels about Heritage and working with IS Oxford:
What are your favourite features in Heritage?
We like the system's efficiency. We like the loans management system. And we like the features of the Online version. We are impressed with its adaptability to the various problems we throw at it. We are impressed with its searching facilities, both ease of simple searching, and being able to select fields from a drop down list. Also the benefits of phrase searching. We are pleased with the ability to create symbolic fields to allow for our demanding search requirements. We like the ease of being able to amend and modify existing records.
Have you linked Heritage to any other systems in your institution, such as Moodle, Sharepoint, Aquabrowser, MyPC, Txtools, Screenreader software, a management information system or similar? If so, any comments about the integration process or success of the integration?
We use Heritage Online URLs in our skills network sites to link through to searches and individual titles. And we use these URLs in our online newsletters around the firm. We look forward to exploiting reading list and review features in this way too.
How have you found the support service from IS Oxford staff?
Always helpful and prompt. And especially helpful in developing outside the box solutions to our requirements.
Have you used the support website? How useful did you find it?
I like the new design, and appreciate the ability to log calls via the website.
Do you have any other comments you would like to make about IS Oxford or Heritage? Would you recommend it/us to other libraries?
We don't hesitate to recommend Heritage to other libraries.
Arup currently have ten additional full access licences, the Serials Management Module (including Acquisitions), the Reader Import Module and the Server Performance Pack. Their data was converted from Inmagic DB/Textworks by IS staff. Heritage Online is used both for internal purposes and to publish the data for the Construction and Building Abstract service run called CBAWeb. Use of Heritage has also extended to their Hong Kong offices.
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