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You can now invite anyone to meet at your favorite Starbucks.
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31 year old man | New York, NY, USA | Seeking women 25-30 within 50 miles
Open minded, spontaneous, always seeking a new experience and rarely pass up on an opportunity explore something new.
I am an experience seeking individual who rarely passes up on an opportunity of exploring something new. I can be very adventurousness but also poses a laid back attitude. I can take the lead when needed but know when to lay low. Like most, I enjoy to travel for various reasons. I enjoy exploring new cities, cultures, customs. I enjoy taking a vacation that strictly consists of laying on the beach for a week. I enjoy adventure filled getaways consisting of skiing, snowboarding, whitewater wafting, mountain biking, etc. I enjoy getting away for a weekend for some peace an quite. I enjoy spending most of time outdoors. Strolling the streets of the city, spending the evening in a lounge, watching a movie or just going with flow and seeing where the evening takes me. (those usually turn our great). I consider myself as a genuinely nice person with solid morals and ethics. A gentleman if you may (grew up in a family of mostly women. They taught me well. :)). I am a true believer in balance. Balance between work and fun, balance in a relationship, balance in all aspects of life. I'm looking for someone that can take me outside of my comfort zone. Someone that can teach me a thing or two about anything. Someone that is open minded and enjoys being spontaneous. More importantly, I'm looking for someone that is understanding, supportive and independent. Someone that has an opinion and not afraid to express it when others don't agree. Someone that can hold their own. No other way to say it but I am looking for someone that is not clingy. I believe that it is possible to have a strong relationship and allow for individual space. I think it all about balance. (I've seen so many people fall of the face of the earth when they meet someone, get married, etc, I certainly don't want to be one of those people. haha) I'm basically looking for my other half (yea, I stole that line).
Camping , Coffee and conversation , Business networking , Cooking , Dining out , Movies/Videos , Museums and art , Music and concerts , Exploring new areas , Nightclubs/Dancing , Shopping/Antiques , Travel/Sightseeing , Watching sports , Wine tasting
Basketball, Billiards / Pool, Bowling, Cycling, Dancing, Football, Golf, Inline skating, Other types of exercise, Running, Skiing, Soccer, Swimming, Tennis / Racquet sports, Walking / Hiking, Weights / Machines, Yoga
I exercise 5 or more times per week.
Middle of the Road
I love the beach and some day will own a beachfront property, preferably in Costa Rica. :) I enjoy everything New York has to offer. The high-line on the west side, promenade in battery park city, strolling the street of Soho in the summer, etc
I have love hate relationship with my phone. Like most others, I am so dependent on it and its certainly a useful tool but at times I just want to shut it off. I enjoy food and prefer to try something new when ever I have the chance.
I enjoy museums, art gallerias, attending sporting events, strolling the streets on New York, attending open houses just for fun, central park, shopping or window shopping, going to random shows at the Javits center (cars, boat, tech shows)
I mostly enjoy reading fiance related material. Weather its magazines or books, I tend to stick with material that I can benefit from. I just re read Jims Cramers Stay Mad For Life.
Z{{Cta}} {{#if HasMoreInfo}} ?{{CtaSecondary}} {{/if}}
|What he is looking for
|6'0" (182cms)
|5'0" (152cms) to 6'0" (182cms)
|Body type:
|Athletic and toned
|Slender, About average, Athletic and toned
|Black, Blue, Brown, Green
|Dark blonde
|Black, Dark brown, Dark blonde
|No Way
|No preference
|Social Drinker
|Social Drinker
|Sales / Marketing
|No preference
|I'll tell you later
|No preference
|Never Married
|Never Married
|Have kids:
|Wants kids:
|White / Caucasian
|Latino / Hispanic, White / Caucasian, Other
Although I certainly do look like it, I am of Armenia decent.
|I'll tell you later
|No preference
|English, Russian
|Bachelors degree
|Bachelors degree, Graduate degree
I have a Bachelors degree in Fiance, Financial Services. I am currently in the process of obtaining a CFA certification. (For those not familiar, ill tell you later.)


text:
Originally written on Boston Sports Media Watch | Last updated 1/2/13
Celtics (29-9) vs. Rockets (17-20) January 2, 1980 The Summit The Celtics finally put together two wins in a row during their six-game road trip, defeating the Rockets, 111-103, at the Summit in Houston. Cedric Maxwell led the C's with 29 points and 10 boards (5 offensive), and his defense was an essential factor in limiting the effectiveness of Houston's brute forward, Moses Malone. Malone still managed to collect 17 rebounds — 11 off the offensive glass — but was limited by Maxwell and Dave Cowens in the post to only 13 points and shot just 31 percent from the field. The win was anything but easy for the Celtics, having already blew leads of 12 and 9 points throughout the game. Bob Ryan recapped the game in the Globe and detailed a furious Celtic rally to win the game: You know how good this one was? Bill Fitch even liked it. Since he usually analyzes each Celtic performance with all the humor of your friendly IRS man in the midst of an audit (". . . Now, about this deduction ...
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text:
ROME, Ga. (AP) There's a new look to the NAIA championship.
RENO, Nev. (AP) Malik Story scored 19 points and Nevada never trailed in beating Cal Poly 69-56 Tuesday night.
SAN LUIS OBISPO, Calif. (AP) Dylan Royer and Chris Eversley scored 15 points each to lead Cal Poly to an 89-43 win against Menlo College on Saturday night.
HONOLULU (AP) Fresno State says it has accepted an invitation to play SMU in the Hawaii Bowl in Honolulu on Christmas Eve.
MORAGA, Calif. (AP) Brad Waldow scored 17 points, leading five St. Mary's players in double digits, and the Gaels beat Cal Poly 86-68 Saturday night.
Waldow was 8 of 9 from the field.


text:
Lesnar Featured On SummerSlam DVD Cover, WWE Has No Swagger Merchandise[Previous Article] - [Next Article] - [Contact Author]
- An image of Brock Lesnar growling is featured on the cover of WWE's upcoming SummerSlam DVD and Blu-ray release on Sept. 18. The cover is the same graphic used for the pay-per-view event's promotional poster.
WWE has been building up a match between Lesnar and Triple H and it will headline SummerSlam on August 19, according to F4WOnline.com.
- Jim Ross responded "Total BS" to a Twitter message Wednesday that WWE was replacing Michael Cole with "Good Ol' J.R."
- WWE currently has no Jack Swagger merchandise in stock. WWEShop.com's page on the former World Heavyweight Champion does not have a single item up for sale. "The All-American American" has struggled greatly since the night after WrestleMania XXVIII, having won only one televised match and losing twenty.


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The Two-Way
8:17 am
Fri January 31, 2014
Top Stories: South Recovers From Weather; Microsoft's Next CEO?
Good morning, here are our early stories:
-- Brain Surgeon Walks Six Miles Through Storm To Save Patient.
-- Reports: Insider Satya Nadella Likely To Be Microsoft's CEO.
And here are more early headlines:
British Nuclear Reprocessing Plant Finds Higher Radiation Levels. (BBC)
Report: Bloomberg To Be Named U.N. Climate Change Envoy. (Reuters)
Violence Feared In Thailand's Election On Sunday. (Reuters)
Iraqi Troops Retake Government Building, Free Hostages. (Al Arabiya)
Colorado Movie Shooting Suspect In Court Today For Hearing. (AP)
Another Cruise Ship Returns To Texas Port With Ill Passengers. (Houston Chronicle)
Short On Cash, New Mexico Is Selling Buffalo Soldiers' Landmark. (AP)
Copyright 2014 NPR. To see more, visit http://www.npr.org/.


text:
Bed-making is a breeze with the Zip Bed
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Article Summary
Italian furniture design company Florida has created the perfect bed for those mornings when you just want to grab a coffee and run out the door. All that you have to do is simply get out and zip it up – no more making your bed in the morning. At night, you just unzip it and climb back in. If you like the snugged-up feeling, you can even be "zipped-in."
Read the full article: Bed-making is a breeze with the Zip Bed
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text:
Bath and Body Works Winter Candy Apple
102 reviews
75% would repurchase
Package Quality: 4.1
Price: $$
Package Quality: 4.1
Price: $$
on 3/27/2015 5:02:00 PM
Age: 25-29
Skin: Other, Fair, Not Sure
Hair: Other, Wavy, Other
Eyes: Other
I wanted to like this but it just smells very gross to me. I pick up a very strong orange tone... I'm not the biggest fan of orange but this smells like a plastic-y orange. The other tones are kind of sugary sweet, which don't really help IMO. Wouldn't use again.
on 3/2/2015 7:26:00 AM
Age: 36-43
Skin: Other, Medium Brown, Not Sure
Hair: Black, Other, Other
Eyes: Black
Beautiful scent!! Its so clean and refreshing I get compliments all the time with this!! Recommended for anyone who wants to smell fresh. If you want this to be sweet add a vanilla based lotion to it. Perfect.
on 2/14/2015 11:17:00 PM
Age: 30-35
Skin: Dry, Medium Brown, Not Sure
Hair: Black, Other, Other
Eyes: Hazel
I would not recommended it was ok
on 1/31/2015 5:43:00 PM
Age: 30-35
Skin: Very Oily, Medium, Neutral
Hair: Brunette, Straight, Fine
Eyes: Brown
I finally tried this scent and was not a big fan. I found it to be pretty boring.
Sweet in a candy way, with a hint of cinnamon like in an air freshner. It just has a strong spiciness to it I do not like.
Just not happy about this scent I will not be purchasing this in any form.
on 1/3/2015 3:15:00 AM
Age: 19-24
Skin: Normal, Fair-Medium, Not Sure
Hair: Blond, Other, Other
Eyes: Blue
My favorite scent from bath and body works!! Not just for the holidays but it works all year round
on 5/20/2014 11:00:00 PM
Age: 36-43
Skin: Very Oily, Fair-Medium, Neutral
Hair: Brown, Curly, Fine
Eyes: Grey
I love the holidays for the scents they bring out. I love apple fragrances & this did not disappoint. If you like Beautiful Day, you may like this one a lot! Apple is hard to get "right" but they managed to make it very sweet but not overpowering. It's simply lovely!! I really enjoy the holiday scents. If you wanted a lovely but somewhat generic, forgettable B&BW scent, this would fit the bill.
on 5/20/2014 4:57:00 PM
Age: 18 & Under
Skin: Normal, Fair, Not Sure
Hair: Black, Other, Other
Eyes: Brown
I like sweet, fruity scents, but this one is just weird. It might be just me because the people around me all like the scent, but I think theres too much of a spice to it; its more harsh than the normal fruit smell. Overall, i dont really like this scent, but i will use this bottle.
on 1/11/2014 11:52:00 AM
Age: 25-29
Skin: Combination, Fair, Cool
Hair: Brunette, Wavy, Medium
Eyes: Brown
I LOVE apple scents but I don't pick up any kind of apple in this. I received a body wash for Christmas in this scent.. it is just soo OVERLY sweet, like a generic sweet smell. Definitely my least favorite holiday body scent that they have released.
on 6/24/2013 6:14:00 PM
Age: 18 & Under
Skin: Acne-prone, Other, Not Sure
Hair: Black, Other, Other
Eyes: Grey
literally the best thing i have ever smelled soooo great i give it 5 stars and recommend it to everyone! it is long lasting sweet smelling and lovely
on 4/1/2013 5:05:00 PM
Age: 30-35
Skin: Sensitive, Fair-Medium, Not Sure
Hair: Brunette, Other, Other
Eyes: Blue
Bath and Body Works changed the scent of this fragrance a few years ago. The original Winter Candy Apple smelled AMAZING! When I realized they had changed it up, I stocked up on the old stuff on ebay (and still have just a few left). But the newer Winter Candy Apple is just too sweet and no longer has that hint of cinnamon or whatever it is that keeps it from smelling overly sweet.
Popular Fragrances
1372 Reviews
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text:
The use of NF-κB inhibitors to enhance the cytotoxic effects of chemotherapy or radiation therapy in the treatment of neoplastic conditions is described.
1. A method of enhancing the cytotoxic effects of radiation therapy, comprising administering to a mammalian subject in need of said radiation therapy a therapeutically effective amount of NF-κB inhibitor in conjunction with the administration of a dosage of the radiation therapy, whereby the cytotoxic effect of said radiation therapy is increased compared to that which would occur in the absence of NF-κB inhibitor;
wherein the dosage of said radiation therapy is decreased as compared to a dosage effective without administering the NF-κB inhibitor in conjuction with the administration of the radiation therapy,
and wherein said subject is afflicted with lung, breast, prostate, brain, kidney, liver, spleen, pancreas, bone or muscle cancer, leukemia or lymphoma.
2. The method according to
3. The method according to
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This application is a continuation of U.S. patent application Ser. No. 10/979,403, filed Nov. 2, 2004, which in turn is a continuation of U.S. patent application Ser. No. 08/959,160, filed Oct. 28, 1997, now issued as U.S. Pat. No. 6,831,057, the disclosures of both of which are incorporated by reference herein in their entirety.
This invention was made with government support under Grant Numbers AI35098 and 1F32-CA69790-01 from the National Institutes of Health and Grant Number DAMD17-94-J-4053 from the Department of the Army. The US Government has certain rights to this invention.
The inducible transcription factor Nuclear Factor-kappa B (NF-κB) participates in the regulation of multiple cellular genes, including many involved in the immune and inflammatory processes (for example, GM-CSF, IL-6, IL-8 and IL-2). NF-κB is a member of the rel family of protein complexes, and is activated by cellular exposure to various factors, including phorbol 12-myristate 13-acetate (PMA), lipopolysaccharide (LPS), interleukin-1 (IL-1), tumor necrosis alpha (TNFα), and ultraviolet radiation. See Baldwin, Annu. Rev. Immunol. 14:649 (1996). NF-κB has also been implicated in the transcriptional activation of several viruses, including HIV-1 (Nabel et al., Nature 326:711 (1987); Kaufman et al., Mol. Cell. Biol., 7:3759 (1987)). The various signaling pathways that can control the activation of NF-κB are not all clearly understood. It is apparent that different inducers can initiate their pathways through distinct receptors.
The latent cytoplasmic form of NF-κB is associated with a cytoplasmic inhibitory protein called IκB. Baeuerle and Baltimore, Science 242:540 (1988); PCT US92/04073 (WO 92/20795). The release of NF-κB from IκB results in the rapid appearance of the active form of NF-κB in the cell nucleus. Genes regulated by NF-κB can be transcriptionally activated minutes after exposure of the cell to the appropriate inducer. Activation of NF-κB after exposure of a cell to an inducer is correlated with the hyperphosphorylation of IκBα and its subsequent degradation. As IκB is diminished in the cytoplasm, NF-κB increases in the nucleus. Phosphorylation of IκB was once thought to lead to dissociation from NF-κB and subsequent proteolysis of IκB. Beg and Baldwin, Genes Dev. 7:2064 (1993). More recently, it has been proposed that the proteasome is responsible for signal-mediated degradation of IκBα and IκBβ. Baldwin, Annu. Rev. Immunol. 14:649 (1996). Phosphorylation of IκB apparently renders the molecule susceptible to proteolysis.
Activation of NF-κB has been suggested as playing a pathological role in the development of autoimmune diseases and chronic inflammatory diseases such as rheumatoid arthritis, and in acute situations such as septic shock.
A first aspect of the present invention is a method of enhancing the cytotoxic effects of an antineoplastic chemotherapeutic agent. A therapeutically effective amount of NF-κB inhibitor is administered in conjunction with the chemotherapeutic agent, so that the cytotoxic effect of the chemotherapeutic agent is increased compared to that which occurs in the absence of NF-κB inhibitor.
A further aspect of the present invention is a method of enhancing chemotherapeutic cytotoxicity in a subject treated with a chemotherapeutic agent. A therapeutically effective amount of NF-κB inhibitor is administered to a subject in conjunction with a chemotherapeutic agent. The cytotoxic effect of the chemotherapeutic agent is increased compared to that which occurs in the absence of NF-κB inhibitor.
A further aspect of the present invention is a method of enhancing the cytotoxic effect of TNFα, by administering a therapeutically effective amount of NF-κB inhibitor in conjunction with TNFα. The cytotoxic effect of TNFα is increased compared to that which occurs in the absence of NF-κB inhibitor.
A further aspect of the present invention is a method of enhancing chemotherapeutic cytotoxicity in a subject treated with TNFα, by administering a therapeutically effective amount of NF-κB inhibitor in conjunction with TNFα. The cytotoxic effect of TNFα is increased compared to that which occurs in the absence of NF-κB inhibitor.
A further aspect of the present invention is a method of screening a compound for the ability to reduce the anti-apoptotic protective effects of an NF-κB induced anti-apoptotic gene. A population of test cells is exposed to an anticancer treatment and a test compound, and the cell viability following the exposure is assessed. Reduced cell viability, compared to that which occurs in cells treated only with the anticancer treatment, indicates that the test compound reduces the anti-apoptotic effects of an NF-κB induced anti-apoptotic gene.
The foregoing and other objects and aspects of the present invention are explained in detail in the specification set forth below.
Many cells are resistant to stimuli which are otherwise capable of inducing apoptosis, but the mechanisms involved are not fully understood. It is demonstrated herein that tumor necrosis factor (TNF), ionizing radiation, and anti-cancer chemotherapeutic compounds (such as daunorubicin), activate NF-κB and that this activation of NF-κB protects the cell against the cytotoxic effects of these treatments. By inhibiting NF-κB nuclear translocation, the present inventors enhanced the apoptotic killing achieved by these reagents; no such enhancement of apoptosis by NF-κB inhibitors was seen with apoptotic stimuli that do not activate NF-κB. These results indicate that NF-κB activation provides cellular resistance to killing by some apoptotic reagents, and provide a method to improve the efficacy of various cancer therapies.
Observations that NF-κB is activated by certain apoptotic stimuli suggests that this transcription factor plays a role in mediating aspects of programmed cell death. Baldwin, Annu. Rev. Immunol. 14:649 (1996); Verma et al., Genes Dev. 9:2723 (1995); Siebenlist et al., Annu. Rev. Cell Biol. 10:405 (1994); Liou and Baltimore, Curr. Opin. Cell Biol. 5:477 (1993); Baeuerle and Henkel, Annu. Rev. Immunol. 12:141 (1994). PCT patent application WO 97/30083 describes anti-apoptotic proteins capable of inhibiting NF-κB, and describes methods of decreasing apoptosis by administering such anti-apoptotic proteins. An anti-apoptotic function of NF-κB is also suggested by the finding that mice lacking the NF-κB p65/RelA gene die embryonically from extensive apoptosis within the liver. Beg et al., Nature 376:167 (1995). Furthermore, it has been observed that many cells which respond to TNF (a strong activator of NF-κB) are resistant to cell killing, and that TNF killing is enhanced in the presence of protein synthesis inhibitors. Holtmann et al., Immunobiology 177:7 (1988).
The present inventors investigated whether activation of NF-κB is protective against the apoptotic killing induced by TNF in a model cell system. Their studies used the human fibrosarcoma cell line HT1080 (which is relatively resistant to killing by TNF; Wang and Baldwin, unpublished data). In order to potentially block the activation of NF-κB in response to TNF stimulation, an HT1080 cell line (HT1080I) expressing a super-repressor form of the NF-κB inhibitor IκBα was established. The super-repressor IκBα contains serine to alanine mutations at residues 32 and 36, which inhibit signal-induced phosphorylation and which inhibit subsequent proteasome-mediated degradation of IκBα. This mutant IκBα protein acts as a super-repressor because it binds to NF-κB and inhibits DNA binding as well as nuclear translocation, but is unable to respond to cellular signals such as those induced by TNF. See Brockman et al., Mol. Cell. Biol. 15:2809 (1995); Brown et al., Science 267:1485 (1995); Traenckner et al., EMBO J. 14:2876 (1995); DiDonato et al., Mol. Cell. Biol. 16:1295 (1996); Palombello et al., Cell 78:773 (1994); Chen et al., Genes Dev. 9:1586(1995). A control line (HT1080V) was established that contained the empty vector and the hygromycin selectable marker. TNFα-induced NF-κB activation, as measured by DNA binding of nuclear extracts, was effectively blocked by the super-repressor IκBα in HT1080I cells as compared to the control cell line (see
As shown in
To exclude the possibility that the expression of the super-repressor form of IκBα leads to a function that is different from the inhibition of NF-κB, the present inventors confirmed the requirement for NF-κB in the inhibition of TNF-induced apoptosis. The pre-treatment of HT1080V cells (control cells) with interleukin-1 (IL-1, an activator of NF-κB that does not initiate apoptosis) blocked the subsequent killing of these cells induced by combined cycloheximide (CHX) and TNF treatment. As shown in
The present inventors additionally determined that proteasome inhibitors enhanced cell killing of HT1080 cells in response to TNF treatment. The degradation of IκB that results in the release of active NF-κB is controlled by the proteasome following inducible phosphorylation and subsequent ubiquitination of IκB (Palombello et al., Cell 78:773 (1994); Chen et al., Genes Dev. 9:1586(1995)). Proteasome inhibitors of the peptide aldehyde category are potent inhibitors of NF-κB activation (Palombello et al., Cell 78:773 (1994); Chen et al., Genes Dev. 9:1586(1995)). The present inventors demonstrated that, in a dose-dependent fashion, the proteasome inhibitor MG132 (Z-leu-leu-leu-H) strongly enhanced the killing of HT1080V cells in response to TNF (data not shown).
If NF-κB inhibition sensitizes cells to TNF killing, then overexpression of NF-κB subunits should restore protection against cell killing in the present HT1080I model. Vectors encoding the p50 and ReIA/p65 subunits of NF-κB or the empty CMV vector control were transfected into the HT1080I cells, and the cells were stimulated with TNT. As expected, the vector alone did not provide protection against cell killing induced by TNF (
Slater et al. reported that in thymocytes (which constitutively express NF-κB) the intensity of the NF-κB/rel protein-DNA complex was enhanced after exposure to etoposide (an agent known to induce apoptosis in thymocytes). Slater et al., Biochem J. 312:844 (1995). These authors concluded that NF-κB expression is correlated with subsequent apoptosis. Bessho et al. examined the effect of pyrrolidine dithiocarbamate (PDTC) on the induction of apoptosis by a variety of agents. Bessho et al., Biochem, Pharm. 48:1883 (1994). They reported that treatment of a human promyelocytic leukemia cell line with etoposide or 1-beta-D-arabinofuranosylcytosine induced NF-κB activation, with apoptosis occurring in 3-4 hours. Addition of PDTC was reported to block NF-κB activation; these authors concluded that etoposide- and dexamethasone-induced apoptosis in human thymocytes was also blocked by PDTC, and that the activation of NF-κB plays a role in the apoptotic process of human hematopoietic cells.
Many cancer therapies function to kill neoplastic cells through apoptotic mechanisms; cellular resistance to apoptosis provides protection against cytotoxic therapies (reviewed in Fisher, Cell, 78:539 (1994)). To determine if other apoptotic stimuli in addition to TNF activate NF-κB and induce a protective effect of NF-κB, the present inventors analyzed ionizing radiation-, daunorubicin- and staurosporine-treated cells. Ionizing radiation has been shown to activate NF-κB in several cell types (Singh and Lavin, Mol. Cell. Biol. 10:5279 (1990); Brach et al., J. Clin. Invest. 88:691 (1991)). The present inventors first investigated whether ionizing radiation, the chemotherapeutic compound daunorubicin, or staurosporine activated NF-κB in the HT1080V cells and in the HT1080I cells. Data shown in
The present inventors also tested whether ionizing radiation-, daunorubicin-, and staurosporine-induced cell killing could be enhanced by the inhibition of NF-κB activity. Daunorubicin (
The present data indicate that the activation of NF-κB by TNF, ionizing radiation, and chemotherapeutic agents (such as daunorubicin, vincristine and irinotecan) protects cells against the apoptotic cell killing induced by these stimuli. Distinct signaling pathways initiated by TNF engagement of its receptor lead to activation of both apoptosis and NF-κB, and NF-κB does not play a positive role in the induction of apoptosis (Tartaglia and Goeddel, Immunol. Today 13:151 (1992); Vandenabeele et al., Trends Cell. Biol. 5:392 (1995); Hsu et al., Cell 81:495 (1995); is Hsu et al., Cell 84:299 (1996); Chinnaiyan et al., Cell 81:505 (1995); Goncharov et al., Cell 84:803 (1996); Hsu et al., Immunity 4:387 (1996)). In the case of ionizing radiation and daunorubicin the activation of apoptosis appears to be initiated by ceramide production (Bose et al., Cell 82:405 (1992); Jaffezou et al., EMBO J. 15:2417 (1996); Haimovitz-Freedman et al., J. Exp. Med. 180:525 (1994); Santana et al., Cell 86:189 (1996)), and the cytotoxic effects of TNF have been reported to require ceramide activation (Kolesnick and Golde, Cell 77:325 (1994)). Ceramide alone has been shown to lead to apoptosis (Obeid et al., Science 259:1769 (1993)) but the details of this apoptotic pathway are not fully understood. It should be noted that several groups have suggested that NF-κB may function pro-apoptotically under some conditions and in certain cell lines (Jung et al Science 268:1619 (1995); Lin et al., J. Cell Biol. 131:1149 (1995); Grimm et al., J. Cell Biol. 134:1 (1996)).
Growing evidence indicates that a variety of anticancer agents kill through programmed cell death. Resistance to anticancer therapies appears to be mediated by resistance to apoptosis (Fisher, Cell 78:539 (1994)). The present data demonstrate that several anticancer agents may be less effective at inducing programmed cell death because of their concomitant activation of NF-κB.
As used herein, "NF-κB activation" refers to NF-κB-mediated up-regulation of genes which are directly or indirectly under the control of an NF-κB binding site. In functional terms, NF-κB activation comprises the binding of NF-κB to κB regulatory sequences in the DNA of a cell, so that transcription of the operatively associated gene is induced (other factors acting in combination with NF-κB may be required to initiate transcription).
"NF-κB inhibition" refers to the prevention of NF-κB binding to NF-κB binding sites in nuclear DNA. NF-κB inhibition does not necessarily imply that all NF-κB-induced transcription is prevented, but rather that such transcription occurs at levels substantially less than that which would occur in the absence of NF-κB inhibition. For example, NF-κB inhibition may result in a 30% reduction in NF-κB-induced transcription of a gene, more preferably a 40%, 50%, or 60% reduction in NF-κB-induced transcription of a gene, and even more preferably a 70%, 80%, 90%, 95% or greater reduction in NF-κB-induced transcription of a gene.
An "NF-κB inhibiting substance", "NF-κB inhibitor", or "NF-κB induction inhibitor" refers to a substance capable of preventing or reducing NF-κB binding to NF-κB binding sites in nuclear DNA, compared to that which would occur in the absence of the NF-κB inhibiting substance.
According to the present invention, methods or compounds that inhibit activation or nuclear translocation of NF-κB, are utilized to enhance the cellular apoptosis caused by various anti-cancer therapies. Methods according to the present invention include treatment of a subject by the administration of an NF-κB inhibitor in conjunction with an anti-cancer therapy, where the anti-cancer therapy alone would induce the anti-apoptotic effects of NF-κB. Addition of the NF-κB inhibitor permits or enhances the apoptotic cell-killing effects of the anti-cancer therapy. Such combined therapy is useful in the treatment of tumors and other neoplastic growths and conditions. Thus, combined therapy that inhibits NF-κB function in the presence of apoptotic stimuli lowers the apoptotic threshold of neoplastic cells and provides a more effective treatment against resistant forms of cancer. Additionally, the inhibition of NF-κB function in association with TNF treatment enhances the previously limited usefulness of this cytokine as an anti-tumor agent.
Methods of the present invention comprise the co-administration of an anti-cancer agent or treatment that induces NF-κB activation, and an NF-κB inhibitor. Such concurrent therapy enhances the cytotoxic ability of the anti-cancer agent, compared to that which would occur in the absence of NF-κB inhibitor. Any suitable NF-κB inhibitor or method of inhibiting NF-κB activation may be used in the present methods. One such method of inhibition is the delivery of the super-repressor IκBα to cells undergoing anti-cancer treatment, including via delivery of nucleotide sequences encoding the super-repressor IκBα. A variety of agents (examples discussed below) are known as NF-κB inhibitors, including proteasome inhibitors, inhibitors of ubiquitin conjugation, inhibitors of proteasome peptidases, and protease inhibitors. Additionally, the use of antisense oligonucleotides to control the expression of cellular components is known in the art, and may be utilized in the present methods to reduce the expression of NFκB or its subunits. (Antisense oligonucleotides that hybridize to NF-κB mRNA, and their therapeutic use to suppress processes that depend on activation of NF-κB, are described in WO95/35032).
One proteolytic pathway in the cytosol involves the covalent conjugation of cellular proteins with the ubiquitin polypeptide (see, e.g., Hershko et al., Ann. Rev. Biochem. 61:761 (1992)). The conjugated proteins are then hydrolyzed by a proteolytic complex containing a degradative particle called the proteasome (Goldberg et al, Nature, 357:375 (1992)). The proteasome is composed of multiple subunits and contains at least three different peptidases (Goldberg et al., Nature 357:375 (1992); Goldberg, Ear. J. Biochem. 203:9 (1992); Orlowski, Biochemistry 29:10289 (1990); Rivett et al., Arch. Biochem. Biophys. 218:1 (1989); Tanaka et al., New Biol. 4:1 (1992)). These peptidases are referred to as chymotrypsin-like peptidase, the trypsin-like peptidase, and the peptidylglutamyl peptidase.
WO 95/25533 describes a method of reducing the cellular content and activity of NF-κB by contacting cells with inhibitors of proteasome function or ubiquitin conjugation; inhibition of proteolysis is achieved by interfering with the ubiquitin dependent pathway at any of several possible steps (e.g., interfering with the activity of the proteasome complex, or interfering with the activity of a proteasome component). Various inhibitors of ubiquitin conjugation to proteins are also known (Wilkinson et al., Biochemistry 29:7373 (1990)).
Various inhibitors of the peptidases of the proteasome have been reported (Dick et al., Biochemistry 30:2725 (1991); Goldberg et al., Nature 357:375 (1992); Goldberg, Eur. J. Biochem. 203:9 (1992); Orlowski, Biochemistry 29:10289 (1990); Rivett et al., Arch. Biochem. Biophys. 218:1 (1989); Rivett et al., J. Biol. Chem. 264:12215 (1989); Tanaka et al., New Biol. 4:1 (1992)). These inhibitors include known inhibitors of chymotrypsin-like and trypsin-like proteases, and inhibitors of thiol (or cysteine) and serine proteases. Some endogenous inhibitors of proteasome activities have also been isolated (Murakami et al., Proc. Natl. Acad. Sci. USA 83:7588 (1986); Li et al., Biochemistry 30:9709 (1991); Golberg, Eur. J. Biochem. 203:9 (1992); Aoyagi et al., Proteases and Biological Control, Cold Spring Harbor Laboratory Press, pp. 429 (1975); Siman et al., WO 91/13904). Potential inhibitors of the chymotrypsin-like activity of the proteasome have been tested Vinitsky et al., Biochemistry 31.9421 (1992); Orlowski et al. Biochemistry 32:1563-1572 (1993); see also WO 95/25533).
Natural and chemical protease inhibitors include peptides containing an α-diketone or an α-keto ester, peptide chloromethyl ketones, isocoumarins, peptide sulfonyl fluorides, peptidyl boronates, peptide epoxides, and peptidyl diazomethanes (see e.g., Angelastro et al., J. Med. Chem. 33:11 (1990); Bey et al., EPO 363,284; Bey et al., EPO 364,344; Grubb et al., WO 88/10266; Higuchi et al., EPO 393,457; Ewoldt et al., Molecular Immunology 29:713 (1992); Hernandez et al., J. Medicinal Chem. 35:1121 (1992); Vlasak et al, J. Virology 63:2056 (1989); Hudig et al., J. Immunology 147:1360 (1991); Odakc et al, Biochemistry 30:2217 (1991); Vijayalakshmi et al., Biochemistry 30:2217 (1991); Kam et al., Thrombosis and Haemostasis 64:133 (1990); Powers et al., J. Cell. Biochem. 39:33 (1989); Powers et al., Proteinase Inhibitors, Barrett et al. (Eds.), Elsevier, pp. 55-152 (1986); Powers et al., Biochemistry 29:3108 (1990); Oweida et al., Thrombosis Research 58:391 (1990)).
Sherman et al. report that pyrrolidine dithiocarbamate (PDTC) is an inhibitor of NF-κB activation. Sherman et al., Biochem. Biophys. Res. Comm. 191:1301 (1993). Anti-inflammatory steroids are suggested as NF-κB induction inhibitors in PCT/US95/12534 (WO/96/10402), including but not limited to predonsone, prednisolone, methyl prednisolone, dexamethasone, prednisone, deoxycorticosterone, cortisone, hydrocortisone. Nonglucocorticoid lazaroids are also suggested for use as NF-κB induction inhibitors. Novel amides that are inhibitors of NF-κB DNA binding are described in WO 97/23457. Japanese patent applications JP 7291859 and JP 7291860 describe chemical inhibitors of NF-κB activity. Additional compounds stated to be useful for inhibiting the action of NF-κB in the nucleus are provided in WO 95/01348. Antisense oligonucleotides that hybridize to NF-κB mRNA, and their therapeutic use to suppress processes which depend on activation of NF-κB, is described in WO95/35032.
The administration of an NF-κB inhibitor to neoplastic cells according to the present invention may also be accomplished by gene therapy, e.g., by transfecting a cell to be treated with a nucleic acid sequence encoding an NF-κB inhibitor. The sequence may be incorporated, according to standard methods, into a mammalian vector suitable for transfecting mammalian cells in vivo, such as an adenovirus, a modified vaccinia virus, an Epstein Barr virus, or the like. The use of retroviral vectors particularly replication-defective retroviral vectors) for gene delivery is well-known in the art and may be used to transform cells for treatment according to the present invention. Alternative methods of targeted gene delivery include DNA-protein conjugates, liposomes, and other methods as are known in the art.
Methods for introducing genes into mammalian cells are known in the art. In the present methods, the vector is introduced to the subject, preferably by direct administration (such as by injection) to the tumor or area to be treated. It is appreciated that the vector may be introduced into nearby non-target cells as well as the target cells when the vector is injected into the treatment area.
Glucocorticoids are widely used as immune and inflammatory suppressants, and inhibit NF-κB (Ray and Prefontaine, Proc. Natl. Acad. Sci. USA 91:752 (1994); Mukaida et al., J. Biol. Chem. 269:13289 (1994); Scheinman et al., Mol. Cell. Biol. 15:943 (1995); Caldenhoven et al., Mol. Endocrinol. 9:401 (1995); Scheinman et al., Science 270:283 (1995); Auphan et al., Science 270:286 (1995)). Glucocorticoids are presently used as part of a therapy for certain hematological malignancies (DeVita et al., Cancer Res, 47:5810 (1987); Schartzman and Cidlowski, Endocrinol. Rev. 14:133 (1993)).
NF-κB inhibitors used according to the methods of the present invention may be administered by any suitable means, as would be apparent to one skilled in the art, including systemically (e.g., intravenously) or locally (e.g., injected into a tumor, a tumor cyst, tissues immediately surrounding a tumor, or into an anatomic compartment containing a tumor). For example, where a therapeutically effective amount of NF-κB is utilized as an adjunct to chemotherapy, the NF-κB may be administered locally to a tumor or neoplasm (or the immediately surrounding tissue). Where a chemotherapeutic agent is delivered systemically, for example, a therapeutically effective amount of NF-κB inhibitor may be administered systemically by intravenous injection. Alternatively, the NF-κB inhibitor may be administered by a route different than that used for the chemotherapeutic agent (e.g., local administration of NF-κB inhibitor to a tumor combined with systemic administration of a chemotherapeutic agent).
The present inventors have determined that anti-cancer treatments can activate NF-κB, and that this activation of NF-κB suppresses apoptosis. The co-administration of NF-κB inhibitors with an anti-cancer treatment, to enhance the apoptotic effects achieved by the cancer treatment, is an aspect of the present invention.
A further aspect of the present invention is a screening assay to identity compounds that inhibit NF-κB induced anti-apoptotic genes (i.e., genes that are induced by NF-κB and that increase the resistance of the cell to apoptotic stimuli, and more specifically, increase the resistance of the cell to the apoptotic effects of an anti-cancer treatment). Such a screening method may comprise exposing a population of test cells to both an anti-cancer treatment and a test compound, and determining cell viability after such exposure. Reduced cell viability or reduced cell survival in the test cells (compared to that which occurs or would be expected to occur after treatment with the anti-cancer treatment alone) indicates that the test compound is capable of inhibiting the anti-apoptotic effects of a gene that is controlled by NF-κB. (The test compound may first be screened to document that it is not an inhibitor of NF-κB activation; any observed reduction in cell survival would thus not be due to inhibition of NF-κB activation, but to effects on the NF-κB induced gene itself.)
A further aspect of the present invention is a method of identifying NF-κB induced genes, including those that have an anti-apoptotic effect. Such a method utilizes techniques known in the art for constructing a cDNA library of a cell, such as polymerase chain reaction (PCR). (See, e.g., U.S. Pat. No. 5,589,622 to Gurr et al.) The cDNA library of a cell exposed to an anticancer therapy that is known to activate the anti-apoptotic effects of NF-κB is compared to the cDNA library of a control cell that was not exposed to the anticancer treatment. cDNA clones representing genes that are induced by activated NF-κB are then identified. The cDNA clones may then be used to isolate the corresponding genomic clone.
Genes induced by NF-κB that suppress apoptosis (i.e., have anti-apoptotic effects) can be used in screening assays to identify compounds that inhibit the anti-apoptotic effects of the genes. Such a method would comprise comparing cells exposed to activated NF-κB and test compound, and cells that were exposed to activated NF-κB only. Compounds that reduce or prevent the anti-apoptotic effects of the identified gene may be useful in enhancing the cytotoxic effects of anticancer therapies, or be useful as anticancer agents themselves.
Thus a further method of the present invention is the co-administration of an anti-cancer treatment with a compound that reduces or prevents the effects of an NF-κB-induced anti-apoptotic gene. Such a compound may act to prevent transcription or translation of the anti-apoptotic gene, or interact with the gene product.
The present invention also provides pharmaceutical compositions for use in the treatment of cancer and neoplastic diseases. The preferred embodiment compositions comprise mixtures of an antineoplastic agent and a suitable NF-κB inhibitor, with suitable pharmaceutical carriers and diluents. Thus, a preferred embodiment composition comprises: (a) an effective amount of an NF-κB inhibitor and (b) an effective amount of a chemotherapeutic agent.
Pharmaceutical compositions in accordance with the present invention may be administered by any suitable means, as would be apparent to one of ordinary skill in the art, including orally (in the form of tablets, capsules or solutions), or parenterally (in the form of injections or pellets). Such preparations can be made using known pharmaceutically accepted carriers and excipients. For administration, the compositions of the present invention will generally be mixed, prior to administration, with a non-toxic pharmaceutically acceptable carrier substance (e.g., normal saline or phosphate-buffered saline), and may be administered using any medically appropriate procedure including oral, intravenous, intraarterial, intradermal, intracavity and intrathecal administration, and direct injection into the tissue to be treated.
Subjects suitable for treatment according to the present invention are preferably mammals, and include humans, non-human primates, and non-primate mammals (including veterinary and livestock subjects), that suffer from a neoplastic disease or tumor, including but not limited to lung, breast, prostate, brain, kidney, liver, spleen, pancreas, bone and muscle cancers. Subjects may have solid or cystic tumors, or diffuse disease.
The methods disclosed herein may be used in subjects having solid or cystic tumors, such as brain tumors or tumors residing in other solid organs. Treatment may comprise administration of NF-κB inhibitors to the tissue of a solid tumor or to 1$ the cystic cavity of a cystic tumor (that is, a tumor surrounding a fluid-filled cavity), or may comprise surgical removal of the solid tumor with administration of NF-κB inhibitor to the resection cavity created by the surgery. Administration of NF-κB inhibitors according to the present invention is in conjunction with chemotherapy or radiation therapy suitable for treatment of the target neoplasm.
Any type of cancer, tumor or neoplasia (both benign and malignant) may be treated by the methods of the present invention, including sarcomas, carcinomas, and mixed tumors. Exemplary conditions that may be treated according to the present invention include fibrosarcomas, liposarcomas, chondrosarcomas, osteogenic sarcomas, angiosarcomas, lymphangiosarcomas, synoviomas, mesotheliomas, meningiomas, leukemias, lymphomas, leiomyosarcomas, rhabdomyosarcomas, squamous cell carcinomas, basal cell carcinomas, adenocarcinomas, papillary carcinomas, cystadenocarcinomas, bronchogenic carcinomas, melanomas, renal cell carcinomas, hepatocellular carcinomas, transitional cell carcinomas, choriocarcinomas, seminomas, embryonal carcinomas, wilm's tumors, pleomorphic adenomas, liver cell papillomas, renal tubular adenomas, cystadenomas, papillomas, adenomas, leiomyomas, rhabdomyomas, hemangiomas, lymphangiomas, osteomas, chondromas, lipomas and fibromas.
According to the present invention, NF-κB inhibitors are utilized as adjuncts in the chemotherapy or radiotherapy of neoplastic disease. The NF-κB inhibitor is provided in a therapeutically effective amount (i.e., an amount sufficient to increase or enhance the cytotoxic effectiveness of the co-administered chemotherapeutic agent or radiation treatment), compared to that which would occur in the absence of NF-κB inhibitor. The use of NFκB inhibitors according to the present method enhances the cytotoxic effects of cancer therapies by preventing activation of NF-κB, thus preventing the protective (anti-apoptotic) effects of NF-κB. Cytotoxicity may be assessed by a reduction in the proliferation of cells and/or decreased numbers of viable cells, leading to a total decrease in the number of viable cells.
As used herein, a therapeutically effective amount of an NF-κB inhibitor refers to those dosages or amounts which decrease or prevent the activation of NF-κB such that the treated cell, or treated population of cells, is more susceptible to the apoptotic effects of a cytotoxic chemotherapy or radiation therapy. Increased susceptibility of cells may be evidenced by decreased cell survival compared to control cells treated without the co-administration of an NF-κB inhibitor; or by the ability to decrease the dosage of the co-administered chemotherapeutic agent or radiation treatment, compared to the dosage required in control cells treated without co-administered NF-κB. Effective dosages of NF-κB inhibitors according to the present invention may vary depending upon the specific NF-κB inhibitor used, the time of administration, the condition being treated, the route of administration, the co-administered cytotoxic agent, and the general condition of the subject being treated. In vitro and animal models to determine particular effective dosages are available and apparent to those of skill in the art. The use of NF-κB inhibiting agents to enhance the cytotoxicity of agents used in the treatment of neoplastic cell growth will be a useful adjunct in treating tumors, cancers, and other neoplastic conditions.
Suitable dosages of NF-κB inhibitors will vary as discussed above, but may range from about 0.001 mg/kg, 0.01 mg/kg, 0.1 mg/kg or 0.3 mg/kg; to about 0.3 mg/kg, 0.5 mg/kg, 1.0 mg/kg, or 10.0 mg/kg, or more.
As used herein, the term chemotherapeutic agent refers to cytotoxic antineoplastic agents, that is, chemical agents which preferentially kill neoplastic cells or disrupt the cell cycle of rapidly proliferating cells, used therapeutically to prevent or reduce the growth of neoplastic cells. Chemotherapeutic agents are also known as antineoplastic drugs or cytotoxic agents, and are well known in the art. As used herein, chemotherapy includes treatment with a single chemotherapeutic agent or with a combination of agents. In a subject in need of treatment, chemotherapy may be combined with surgical treatment or radiation therapy, or with other antineoplastic treatment modalities.
Exemplary chemotherapeutic agents are vinca alkaloids, epipodophyllotoxins, anthracycline antibiotics, actinomycin D, plicamycin, puromycin, gramicidin D, paclitaxel (TAXOL�, Bristol Myers Squibb), colchicine, cytochalasin B, emetine, maytansine, and amsacrine (or "mAMSA"). The vinca alkaloid class is described in Goodman and Gilman's The Pharmacological Basis of Therapeutics, 1277-1280 (7th ed. 1985) (hereafter "Goodman and Gilman"). Exemplary of vinca alkaloids are vincristine, vinblastine, and vindesine. The epipodophyllotoxin class is described in Goodman and Gilman, supra at 1280-1281. Exemplary of epipodophyllotoxins are etoposide, etoposide orthoquinone, and teniposide. The anthracycline antibiotic class is described in Goodman and Gilman, supra at 1283-1285. Exemplary of anthracycline antibiotics are daunorubicin, doxorubicin, mitoxantraone, and bisanthrene. Actinomycin D, also called Dactinomycin, is described in Goodman and Gilman, supra at 1281-1283. Plicamycin, also called mithramycin, is described in Goodman and Gilman, supra at 1287-1288. Additional chemotherapeutic agents include cisplatin (PLATINOL� Bristol Myers Squibb); carboplatin (PARAPLATIN�, Bristol Myers Squibb); mitomycin (MUTAMYCIN� Bristol Myers Squibb); altretamine (HEXALEN�, U.S. Bioscience, Inc.); cyclophosphamide (CYTOXAN�, Bristol Myers Squibb); lomustine [CCNU] (CEENU�, Bristol Myers Squibb); carmustine [BCNU] (BICNU�, Bristol Myers Squibb); irinotecan (CPT-11).
Methods of administering chemotherapeutic drugs vary depending upon the specific agent used, as would be mown to one skilled in the art. Depending upon the agent used, chemotherapeutic agents may be administered, for example, by injection (intravenously, intramuscularly, intraperitoneally, subcutaneously, intratumor, intrapleural) or orally.
As used herein, the administration of a compound "in conjunction with" a second compound means that the two compounds are administered closely enough in time that the presence of one alters the biological effects of the other. The two compounds may be administered simultaneously (concurrently) or sequentially. Simultaneous administration may be carried out by mixing the compounds prior to administration, or by administering the compounds at the same point in time but at different anatomic sites or using different routes of administration.
The phrases "concurrent administration", "simultaneous administration" or "administered simultaneously" as used herein, means that the compounds are administered at the same point in time or immediately following one another. In the latter case, the two compounds are administered at times sufficiently close that the results observed are essentially indistinguishable from those achieved when the compounds are administered at the same point in time.
Many chemotherapeutic agents act at specific phases of the cell cycle, and are active only against cells in the process of division. Neoplasms which are the most susceptible to chemotherapy are those with a high percentage of cells in the process of division, including but not limited to breast, liver, brain, lung, and ovarian cancer.
Anti-cancer radiotherapy may be carried out in any conventional manner as is known in the art, including external beam radiotherapy and implanted radioactive sources (such as temporarily implanted radioactive iodine sources to deliver high dose local therapy).
Applicants specifically intend that the disclosures of all U.S. patents cited herein be incorporated by reference in their entirety
The following examples are provided to illustrate the present invention, and should not be construed as limiting thereof.
HT1080 fibrosarcoma cells were co-transfected with the pCMV empty vector to produce control cells HT1080V; HT1080, cells co-transfected with the pCMV vector containing a cDNA encoding the super-repressor IκBα and with the pCEP4 vector for hygromycin B selection (400 μg/ml) provided HT1080I cells that expressed the super-repressor IκBα. Transfection was by the lipofectamine protocol (Gibco/BRL. IκBα levels were determined by immunoblotting (ECL, Amersham) to equivalent amounts of protein from the different cell lines with an antibody to human IκBα (Rockland, Inc., Royertown, Pa.). The expression of super-repressor IκBα in HT1080I cells efficiently blocked TNF-stimulated NF-κB nuclear translocation as determined by electrophoretic mobility-shift assay (EMSA) (data not shown).
HT1080V (open squares) and HT1080I cells (solid diamonds) were treated with TNFα (20 ng/ml) for varying times and surviving cells were quantified by crystal violet assay (Mehlen et al., EMBO J. 15:2695 (1996)). Data shown (
TNF-induced apoptosis was detected by TUNEL staining (Graeber et al., Nature 379:88 (1996)). HT1080V (V) and HT1080I (I) cells were untreated (−TNF) or stimulated (+TNF) with 50 ng/1 ml TNFα for 7 hr and fixed with 4% paraformaldehyde. The staining was done according to the manufacturer's instructions (Boehringer Mannheim). HT1080I cells treated with TNF showed the condensed morphology typical of apoptotic cells (
To exclude the possibility that the expression of the super-repressor form of IκBα leads to a function that is different from the inhibition of NF-κB, the requirement for NF-κB in the inhibition of TNF-induced apoptosis was confirmed. Cells were pre-treated with interleukin-1 (IL-1), an activator of NF-κB that does not initiate apoptosis, prior to combined cycloheximide (CHX) and TNF treatment.
HT1080V and HT1080I cells were preincubated with 10 ng/ml IL-1β (R & D Systems) for 5 hours (+Pre IL-1), or were left untreated. Cells were then treated with 10 μg/ml cycloheximide (CHX) and different concentrations of TNFα (0-50 ng/ml). Surviving cells were quantified by crystal violet assay (Mehlen et al., EMBO J. 15:2695 (1996)). As shown in
The effect of a proteasome inhibitor on TNF-treated cells was investigated. Proteasome inhibitors of the peptide aldehyde category are potent inhibitors of NF-κB activation (Palombello et al., Cell 78:773 (1994); Chen et al., Genes Dev. 9:1586(1995)). It was demonstrated that, in a dose-dependent fashion, the proteasome inhibitor MG132 (Z-leu-leu-leu-H) strongly enhanced TNF-induced killing of HT1080V cells (data not shown). Cell viability experiments were used to monitor the effectiveness of MG132 in enhancing TNF-induced cell killing.
Expression of the p50 and RelA/p65 NF-κB subunits restored cell resistance to TNF killing in cells expressing the super-repressor IκB. HT1080I cells (expressing the super-repressor IκBα) were either co-transfected with pcDNA3-LacZ (Invitrogen) and pCMV-p65 (2 μg) and pCMV-p50 (2 μg) (hatched bars); or with the LacZ and empty vectors. Transfection was by the lipofectamine protocol. After 40 hours, cells were treated with different concentrations of TNFα for an additional 24 hours. The results are from the mean�SD of two experiments.
As shown in
Daunorubicin and ionizing radiation induce nuclear translocation of NF-κB. HT1080V (V) and HT1080I (I) cells were treated with 1 μM daunorubicin (Sigma), 50-nM staurosporine (Sigma), or were irradiated (5 Gy) for the indicated times. EMSA was performed as previously described (Finco et al., Proc. Natl. Acad. Sci. USA, 91:11884 (1994) to assess nuclear translocation of NF-κB. Results of EMSA are provided in
As shown in
As indicated in
Five hundred HT1080V cells and 500 HT1080I cells were plated in separate six-well plates. Twenty-four hours later the cells were exposed to ionizing radiation in doses from 0 Gy to 10 Gy, and fourteen days later cell clones were counted. Each group was performed in triplicate. Results are shown in
The above results indicate that the overexpression of super-repressor IκBα in HT1080I cells enhanced cell killing by daunorubicin and ionizing radiation.
The present inventors' preliminary data additionally indicate that cell killing by vincristine is augmented by the inhibition of NF-κB (data not shown). HT1080V and HT1080I cells were treated with vincristine and cell viability was determined. Cell killing in HT1080I cells was enhanced compared to that in HT1080V cells.
The antineoplastic agent CPT-11 (irinotecan) was determined to activate NF-κB (data not shown). The co-administration of the super-repressor IκB with CPT-11 was investigated in nude mice with experimentally induced fibrosarcomas. Delivery of the super-repressor IκB was achieved by injecting tumors directly with an adenovirus encoding the super-repressor IκB under the control of the cytomegalovirus (CMV) promoter. A control adenoviral vector contained the CMV promoter but not the coding sequence for IκB. Animals were systemically treated with CPT-11, according to protocols known in the art.
Treatments were as indicated in
As shown in
The above results indicate that the efficacy of anti-neoplastic therapies can be enhanced by the concomitant suppression of NF-κB activation.
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Great looking and instant performance gain
Reviewed By John K (AUBREY, TX) Reviewed for a 1990 Chevy C/K 1500 —7/1/2015 8:18 PM
You could instantly tell the difference and it was simple to install. Put this on my 1990 Chevy 454
Reviewed By Joe M (Framingham, MA) Reviewed for a 2013 Chevy Malibu —7/1/2015 3:20 PM
Great system. Very easy to follow instructions and good design make for easy installation. Cleans up the engine bay nicely. Slight mileage increase on the first tank of gas. Only downsides are a resonant intake noise when you get on it, to be expected when getting rid of the factory intake though and it's a bit pricey.


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Commercial painter in serious condition after being electrically shocked in Jupiter
JUPITER — A 26-year-old commercial painter was listed in serious condition Thursday at St. Mary's Medical Center after being electrically shocked, according to a police news release.
Police say Alex Tobar of Plantation was unresponsive Wednesday, with no pulse and his heart had stopped, before he was revived by two police officers who performed CPR and used an automated external defibrillator until Palm Beach County Fire Rescue arrived. The man was flown to St. Mary's in West Palm Beach.
Officers Janine Jenne and Sally Collins were on foot patrol on Honey Bell Way Wednesday afternoon when they heard a call for help.
"I heard the males scream out: 'he's been electrocuted,' so right away I just resorted right back to my training ... luckily we were right there ... it felt good, it really felt good to see his mom crying with tears of joy at the hospital," said Jenne Thursday afternoon as she recalled the incident in the 100 block of Seagrape Drive, near the intersection of Military Trail and Indian Creek Parkway.
Police say Tobar was working on a painting project inside a bucket lift when he came in contact with power lines.
WPTV NewsChannel 5 contributed to this report
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Reviews for Ford F-250 years:
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Ford F-250 Reviews and Owner Comments
8 Reviews
1992 Ford F-250stp, East Berlin, CT, August 22, 2012 13:04I've got a 1992 5.8L 2WD. I'm the fourth owner and there are only 107k miles. My kids fit in the back seat and I can haul just about anything with this beast. Since I've owned it I've had to replace the battery, replace rear brake lines and the front calipers are frozen and need to be replaced. The steering is a little loud but works fine and the car sometimes locks itself and I learned to leave the triangle windows unlocked to avoid having to break into my own car when I leave the keys in it. Otherwise problem free. This has been a GREAT truck!Enthusiast
1996 Ford F-250Visitor, Batesville, IN, August 10, 2012 07:09My truck just turned 186,000 miles on it and the only thing I've done to it was a new battery, brakes, and tires. It still drives like a dream especially considering the miles and i haul with it several times a week.
1992 Ford F-250Visitor, Crowley, LA, September 11, 2011 09:09I have a 1992 Ford F-250 5.8 and I changed the fuel pump and filter and their is fuel at the fuel rail but it's not going into the throttle body. now when I put gas into the throttle it runs until it runs out what could be the problem?
1997 Ford F-250Visitor, Nicholson, GA, May 15, 2011 13:33fuel system is not pumping fuel from tanks to the fuel filter
1993 Ford F-250Visitor, La Mesa, CA, September 14, 2010 23:14Everything but the engine needs replacing at about 100K miles.
1999 Ford F-250Visitor, Dacula, GA, September 10, 2010 13:42heater core
1996 Ford F-250gkb1000, Hanover, PA, April 24, 2009 12:04Only major problem was Transmission shift problem at 12000 miles. Ford "replaced" tranny (under warranty), started again at 20000 miles. Amaco fixed for $87. parts and labor no problems since now at 77000 milesEnthusiast
1996 Ford F-250blindsquirrel, San Andreas, CA, December 30, 2008 16:34Great Truck! The glow plugs had to be replaced and the water pump replaced because it was leaking. Never left me stranded or worried. I have the 7.3L Turbo DieselJourneyman
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In a television signal processing apparatus, a phase-locked loop (PLL) is provided for detecting the audio information from the television signal and for providing a defect control signal which, when applied to compensation circuitry, reduces impulse noise effects in the video information. The PLL includes a phase detector, a lowpass filter (LPF) and a voltage controlled oscillator (VCO). The phase detector has a first input coupled to be responsive to the frequency modulated sound signal produced by the tuner, a second input coupled to be responsive to the output of the VCO and an output coupled to the LPF. The LPF provides a phase control signal to the VCO and provides the detected audio information. A signal mixer serving as a synchronous detector has first and second inputs coupled to the first and second inputs, respectively, of the phase detector and has an output for providing the defect control signal indicative of the amplitude variations of the frequency modulated sound carrier.
1. In a television system including a source of an intermediate frequency (IF) signal comprising an IF picture carrier amplitude modulated with video information and an associated IF sound carrier frequency modulated with audio information, having processing circuits for detecting said video information, and having compensation circuitry responsive to a defect control signal for reducing effects of noise in said picture carrier, apparatus for detecting said audio information and for providing said defect control signal comprising:
a controllable oscillator responsive to an oscillator control signal coupled to an oscillator control input and having an output at which is provided a controlled frequency output signal;
a frequency translation circuit having a first input coupled to be responsive to said IF picture carrier, a second input coupled to be responsive to said oscillator output signal, and an output for providing a frequency translated signal;
a phase detector having a first input coupled to be responsive to said IF sound carrier, a second input coupled to be responsive to said frequency translated signal, and an output;
lowpass filter means coupled between the output of said phase detector and the control input of said controllable oscillator for providing said oscillator control signal and for providing said audio information;
a synchronous detector having a first input coupled to be responsive to said sound carrier, a second input coupled to be responsive to said frequency translated signal, and an output for providing detected amplitude variations of said sound carrier indicative of the presence of noise in the picture carrier; and
control circuitry coupled to the output of said synchronous detector for providing said defect control signal whenever said detected amplitude variations exceed a predetermined threshold.
2. The apparatus of claim 1 further including:
gain controlled amplifier means coupled between said source of IF signal and said first input of said synchronous detector, said gain controlled amplifier means having a relatively slow gain control time response for maintaining the long term average amplitude of said modulated sound carrier substantially constant, but not substantially reducing the amplitude variations due to noise.
3. The apparatus of claim 2 wherein:
said frequency translation circuit, said phase detector and said synchronous detector each comprise an analog multiplier circuit.
4. In a television system including a source of an intermediate frequency (IF) signal comprising an IF picture carrier amplitude modulated with video information and an associated IF sound carrier frequency modulated with audio information, having processing circuits for detecting said video information, and having compensation circuitry responsive to a defect control signal for reducing effects of noise in said picture carrier, apparatus for detecting said audio information and for providing said defect control signal comprising:
a phase-locked loop having a phase detector, filter means and a frequency controlled oscillator, said phase detector having a first input coupled to be responsive to said frequency modulated sound signal, a second input coupled to be responsive to an output signal of said controlled oscillator and an output coupled to said filter means, said filter means coupling a control signal to said controlled oscillator and providing at an output terminal said audio information; and
means for detecting amplitude variations of said frequency modulated sound carrier, having a first input coupled to be responsive to said modulated sound carrier, a second input coupled to be responsive to said output signal of said controlled oscillator and an output for providing said defect control signal to said compensation circuitry in response to said detected amplitude variations of said frequency modulated sound carrier.
5. The apparatus of claim 4 wherein said means for detecting amplitude variations comprises a synchronous detector.
6. The apparatus of claim 5 further including:
gain controlled amplifier means coupled between said source of IF signal and said synchronous detector, said gain controlled amplifier means having a relatively slow gain control time response for maintaining the long term average amplitude of said modulated sound carrier substantially constant, but not substantially reducing the amplitude variations due to noise.
7. The apparatus of claim 5 wherein said synchronous detector includes:
a mixer having a first input coupled to said first input of said phase detector and a 90� phase shift network coupled between a second input of said mixer and said second input of said phase detector.
8. The apparatus of claim 7 wherein said means for detecting amplitude variations further includes:
a source of substantially constant reference signal; and
a comparator having a first input for receiving said constant reference signal, and a second input for receiving amplitude variations detected by said synchronous detector, said comparator generating an output signal whenever said detected amplitude variations exceed the constant reference signal.
9. The apparatus of claim 8 wherein said means for detecting amplitude variations further includes circuitry, responsive to the comparator output signal, for generating at an output terminal said defect control signal having a predetermined amplitude and duration.
10. The apparatus of claim 4 wherein said phase-locked loop further includes a frequency translation circuit coupled between said frequency controlled oscillator and said second input of said phase detector, said frequency translation circuit being responsive to said IF picture carrier for frequency translating said oscillator output signal to the frequency of said IF sound carrier.
11. The apparatus of claim 10 wherein said frequency translation circuit, said phase detector and said mixer each comprise an analog multiplier.
The present invention relates to television detector circuits and more particularly, to a phase-locked loop (PLL) sound signal demodulator arrangement for detecting the television sound signal and indicating the presence of impulse noise in the video signal.
Television receivers are susceptible to signal interference from various sources. The interference may arise as electromagnetic radiation which interferes with the broadcast RF signal. Typical sources of such interference are automotive ignition systems and household motors such as are found in vacuum cleaners and hair dryers, etc. The interference is manifested as bright or dark spots appearing in the TV picture.
Television pictures are formed by a scanning technique in which the image is partitioned into a plurality of horizontal lines. The horizontal lines are sequentially transmitted to the receiver where they are successively reconstructed into the original image on the picture screen. The signal information from image line to image line is highly redundant for a large percentage of images. As described in U.S. Pat. No. 2,996,576 issued Aug. 15, 1961 in the name of R. Dolby, advantage can be taken of this line-to-line signal redundancy to minimize the effect of TV signal disturbances in video recording/reproducing systems such as video tape recorders and video disk systems. Upon detection of a video signal loss, the signal for the preceding image line, which is delayed by one image line period, is substituted for the lost signal to minimize picture disturbance. Detection of signal loss in such recording/reproducing systems is facilitated by the fact that the video signal is recorded as frequency modulation of a carrier. Thus, video signal loss may be detected by detecting the loss of the FM carrier, i.e., by envelope detection of the FM signal. See for example, U.S. Pat. No. 4,203,134 issued May 13, 1980 in the name of T. J. Christopher, et al.
Relatively large numbers of defects in video recording media makes it imperative to include signal compensation systems in video recording/reproducing apparatus. On the other hand, the infrequency of particularly objectionable interference in broadcast TV signals has until recently made it relatively impractical from a cost point of view to include such defect correction systems in TV receivers for eliminating interference. However, with the incorporation of charge transfer device delay lines in TV receivers to perform comb filtering of the luminance signal, it has become practical to consider correcting video interference due to noise impulses. The interference correction can be performed by video signal substitution as in the recording/reproducing systems. However, the detection of noise or defects in the video signal in the TV receiver presents problems different from defect detection in the recording/reproducing systems.
Typically the interference or noise creating the picture disturbance is not of the type (i.e., sufficient amplitude) to cause elimination of the broadcast signal carrier, thus one cannot rely upon envelope detection of the broadcast signal carrier to determine when defects are present in the signal. In addition, the broadcast signal is an amplitude modulated carrier so that amplitude changes are generally not indicative of the presence of noise or signal defects. That is, since the dynamic amplitude range of the baseband video signal is very large, impulse noise can have signal values from barely perceptible to a level that actually overloads the receiver circuits.
In U.S. Pat. No. 4,353,093 issued Oct. 5, 1982 in the name of J. Durbin et al., it is recognized that impulse noise, i.e., noise signals which may supersede the video signal on all or a part of an image line, typically has a broad energy spectrum and causes interference across many TV channels. Therefore, energy from the impulse is likely to be included in the sound component of a particular TV signal even though the sound component of the TV signal is broadcast on a carrier separate from the video signal component of the particular TV signal. Since the sound component of the TV signal is a frequency modulated waveform, amplitude detection of such a signal can be used for effectively detecting the presence of relatively small values of impulse noise.
In the Durbin et al. patent, a phase-locked loop synchronous detector arrangement is responsive to a bandpass filtered version of the modulated sound carrier for detecting amplitude variations of the sound carrier. Compensation circuitry, including a delay line, coupled to the output of the synchronous detector substitutes a prior image line for the present image line in response to the level of the detected amplitude variations. In addition to this phase-locked loop synchronous detector arrangement for detecting impulse noise, conventional intercarrier detection circuitry is required for detecting the audio and video information from the composite TV signal.
In U.S. patent application Ser. No. 412,902, now Pat. No. 4,470,071, entitled TELEVISION SOUND DETECTION USING A FREQUENCY TRANSLATION PHASE-LOCKED LOOP, filed in the name of Abraham E. Rindal on Aug. 30, 1982 and assigned, like the present application, to RCA Corporation, an audio detection system is described for processing a television signal which may include a multichannel audio signal component. In that system, the IF picture carrier provided at the output of the TV tuner is processed in a separate IF channel for developing the video information. Audio information is detected by a frequency translation phase-locked loop (PLL) including a mixer and a phase detector responsive to the IF picture and sound carriers, respectively, provided at the output of the tuner.
In accordance with the present invention a system is provided which detects video signal noise defects and recovers the audio information from a TV signal with a single phase-locked loop circuit arrangement and, which can easily be constructed in integrated circuit form. More specfically, in accordance with the principals of the present invention, the PLL includes a phase detector, a filter and a frequency controlled oscillator for detecting the audio information. The phase detector has a first input coupled to be responsive to a frequency modulated sound signal coupled from a tuner, a second input coupled to be responsive to the output of the controlled oscillator and an output coupled to the filter. The filter couples a control signal to the controlled oscillator for controlling the frequency of the oscillator output signal and for providing audio information at an output terminal. A means for detecting amplitude variations of the frequency modulated sound carrier is provided, having a first input coupled to be responsive to the modulated sound signal, a second input coupled to be responsive to the output of the controlled oscillator and an output for providing a defect control signal to defect compensation circuitry in response to the detected amplitude variations of the modulated sound carrier.
The sole FIGURE is a block diagram of TV receiver circuitry including a preferred embodiment of the present invention.
Referring to the drawing, a television broadcast signal received by an antenna 8 is applied to a tuner 10. Tuner 10 selectively translates the radio frequency (RF) picture and sound carriers of a selected TV channel to respective intermediate frequency (IF) carriers at, e.g., 45.75 MHz and 41.25 MHz, respectively, in the NTSC system. The IF carriers are available at tuner output terminal 12. The IF picture carrier is basically an amplitude modulated (AM) signal containing composite video information. The IF sound carrier, on the other hand, is a frequency modulated (FM) signal. Conventional color television signal processing circuitry including an IF filtering and amplifying channel 14 and a video detector 16 are responsive to the IF picture carrier at terminal 12 for detecting a composite baseband video signal. The baseband video signal is applied to an input terminal 17 of a comb filter 18 which separates the chrominance (C) and luminance (Y) from the composite baseband video signal and provides respective signals at terminals 19 and 20. A more detailed description of comb filter 18 will be provided later on. The luminance and chrominance signals are processed in conventional fashion to develop color signals which are supplied to respective guns of a kinescope (not shown). The resultant electron beams are deflected in successive horizontal image lines to produce an image on the screen of the kinescope in a conventional manner.
IF channel 14 has a conventional passband response 21 in which the IF picture carrier P is located 6 dB down on the higher frequency slope of passband response 21 and the IF sound carrier S is located approximately 25 dB down on the lower frequency slope of passband response 21. As a result, the vestigial sideband video information can be detected without significant interference from the IF sound carrier. Unfortunately, IF passband response 21 induces what is called a "Nyquist" incidental carrier phase modulation (ICPM) to the IF picture carrier, due to its asymmetrical attenuation of the sidebands of the IF picture carrier. In a conventional intercarrier type of sound detection system this Nyquist induced ICPM of the IF picture carrier results in distortion of the intercarrier sound signal due to the process of mixing the IF sound and picture carriers. This distortion of the intercarrier sound signal results in audio buzz. The amount of audio buzz caused by Nyquist ICPM increases in direct relationship with increases in the bandwidth of the audio signal. Thus, when one considers that the bandwidth of a composite audio signal including multiple subcarriers for the transmission of, for example, monophonic, stereophonic and second audio program signals is significantly greater than the bandwidth of the monophonic signal conventionally processed by an intercarrier detector, it becomes apparent that the conventional intercarrier type of audio detection is likely to be unsatisfactory since the audio buzz produced will be substantially increased.
In the present receiver, a frequency translation phase-locked loop (PLL) 22 process the IF sound carrier in a path separate from the IF picture carrier processing circuitry for detecting the audio information to substantially reduce the Nyquist induced ICPM. A buffer amplifier 24 couples the IF picture and sound carriers from terminal 12 to picture and sound bandpass filters 26 and 28, respectively. Picture bandpass filter 26 has a symmetrical, relatively narrow (e.g., a 3 db bandwidth of 1 MHz) passband response 27 centered at the IF picture carrier frequency P (e.g., 45.75 MHz in the NTSC system) for passing substantially only the IF picture carrier signal. An IF amplifier 30 and a limiter 32 appropriately amplify and limit the IF picture carrier signal and apply the resultant limited, and therefore substantially unmodulated, IF picture carrier to one input of a signal mixer 34.
Sound bandpass filter 28 has a symmetrical, relatively narrow (e.g., a 3 db bandwidth of 1 MHz) passband 29 centered at the IF sound carrier frequency S (e.g., 41.25 MHz in the NTSC system) for passing substantially only the IF sound carrier signal and its immediate sidebands which are applied, after amplification by an IF amplifier 36, as one input to a phase detector 38. IF amplifiers 30 and 36 may be similarly configured and each comprise, for example, an integrated circuit IF amplifier such as the TA 7607 manufactured by Tokyo Shibaura Electric Co., Ltd. Limiter 32 may simply comprise parallel connected, oppositely poled Schottky-barrier diodes. A conventional automatic gain control circuit (AGC) 40 is responsive to the output of IF amplifier 36 (or alternatively to the output of amplifier 30) for supplying AGC control voltages to IF amplifier 30 and 36 for controlling their gain so as to establish their output signals at predetermined levels.
A varactor tuned voltage controlled oscillator (VCO) 42 having a nominal oscillation frequency equal to the frequency difference (e.g., 4.5 MHz for the NTSC system) between the IF picture and sound carriers provides a second input to mixer 34. Mixer 34 may comprise, for example, a double balanced analog multiplier circuit, such as the MC1496 available from Motorola Semiconductor Products, Inc., which operates in a switching mode in response to the amplitude limited IF picture carrier (which acts as the switching control signal) for mixing the IF picture carrier signal and VCO output signal to provide at its output a frequency translated signal representing the frequency and phase differences between its input signals. In the NTSC system, the frequency difference between the input signals of mixer 34 is 41.25 MHz. This 41.25 MHz output signal of mixer 34 is supplied as a second input to phase detector 38, which can also comprise an MC1496 integrated circuit. Phase detector 38 provides an output signal having an amplitide which varies in direct relation with the phase difference of its input signals and thus operates as a frequency modulation (FM) demodulator for the FM modulated IF sound signal coupled to its first input from amplifier 36 and provides at its output a composite baseband audio signal (along with undesired signals which result from the detection process). For a more detailed discussion of the operation of analog multipliers as FM demodulators see an article entitled "Applications of a Monolithic Analog Multiplier" by A. Bilotti, in IEEE Journal of Solid State Circuits, Dec. 1968, pages 373-380.
A lowpass filter 44 coupled to the output of phase detector 38 filters its output signal to provide a control signal for controlling the frequency of VCO 42. Mixer 34, phase detector 38, VCO 42 and lowpass filter 44 comprise the frequency translation phase-locked loop 22. Lowpass filter 44 has a sufficient bandwidth for selecting the composite baseband audio signal but narrow enough for rejecting the higher frequency undesired signals and supplies the composite audio information signal, for example, to a stereo decoder 46. If stereo programming is provided in the composite audio signal, decoder 46 will decode the composite audio signal and provide left and right stereo signals to speakers 48 and 50, respectively. If stereo programming is not provided, decoder 46 will provide the monophonic signal to the speakers.
In operation, mixer 34 frequency translates the IF picture carrier signal applied at its input and provides an output signal having a frequency corresponding to the difference of the frequencies of its input signals (e.g., 41.25 MHz). Undesired output signals corresponding to feedthrough of the input signals and the sum of the input signals are attenuated by the relatively narrow bandwidth of lowpass filter 44. The amplitude of the output voltage of phase detector 38 is a measure of the phase difference between its input signals. This output voltage is filtered and applied by lowpass filter 44 as a control voltage to VCO 42. The frequency of the output signal of VCO 42, and consequently the 41.25 MHz translated signal from mixer 34, changes in direct relation to the amplitude of the control voltage and is in a direction for reducing the phase difference between the signals at the inputs of phase detector 38. When the phase of the input signals of detector 38 are in quadrature (90�), a minimum amplitude control voltage is applied to VCO 42. Therefore, due to the feedback nature of loop 22, when it is locked, the control voltage at the output of filter 44 is such that the frequency of the translated difference signal at the output of mixer 34 is equal to the average frequency of the IF sound carrier and its phase is in quadrature in relation to the phase of the IF sound carrier. Thus, phase detector 38 acts as an FM detector for frequency demodulating the IF sound carrier to produce a signal which, when filtered by filter 44, corresponds to the audio information.
Since mixer 34 is also responsive to the picture carrier signal, any common mode FM which may be imparted to both the picture and sound carrier by, e.g., the local oscillator of tuner 10 or television accessories such as a cable television converter which may precede tuner 10, is transferred by mixer 34 to the frequency translated signal and therefore cancelled in phase detector 38.
As previously noted, the frequency modulated sound carrier is selected from the output of tuner 10 by bandpass filter 28 and amplified by amplifier 36. Amplifier 36, controlled by AGC circuit 40, conditions the sound carrier so as to have a constant long term average amplitude, and tends to remove normally occurring amplitude fluctuation not associated with noise. The time constants of AGC system are not sufficiently fast so as to limit the occurrence of amplitude fluctuations due to impulse noise. Thus, except for impulse noise induced amplitude fluctuations, the amplitude of the modulated sound carrier is substantially constant. Since PLL's are relatively insensitive to amplitude fluctuations, a limiter is not required between amplifier 36 and detector 38 for eliminating the impulse noise fluctuations.
Consequently, the output of amplifier 36 is an advantageous point for detecting amplitude fluctuations of the modulated sound carrier for deriving a defect control signal and for reducing impulse noise induced video signal defects. Additionally, since the bandwidth of filter 28 is wide enough to pass the sound carrier and some of its sidebands, it presents as relatively shorter signal delay than the delays in the video signal path and allows sufficient time for video signal defect compensation. A defect control signal generating circuit 52 including a synchronous detector 54 for detecting the impulse noise induced fluctuations can be advantageously incorporated with the structure of the previously described PLL by the addition of a minimum number of parts and in a manner easily incorporated with the structure of the audio detector PLL, desirably in integrated circuit form.
Since the output signal of mixer 34 has a frequency equal to the average frequency of the frequency modulated sound carrier but has a quadrature phase, a 90� phase shift network 56, shifts the output signal of mixer 34 90� and applies it to one input of synchronous detector 54. Thus, when the FM sound carrier is applied to the other input of detector 54, the phase of its input signals are in synchronism (i.e., in phase alignment) with each other and thus synchronous detector 54, in a well known manner, detects the amplitude variations of the FM sound carrier. Detector 54 may also comprise e.g., a double balanced multiplier such as found in the prior noted MC 1496 integrated circuit.
The amplitude variations produced at the output of detector 54 are applied to a first input terminal of comparator circuit 58. A DC threshold signal from source 60 is applied to a second input of comparator 58. The comparator produces an output state change whenever the output signal of detector 54 passes through the threshold level. The output signal of detector 54 will be a slowly varying AC signal having a relatively small amplitude except when impulse noise or other dramatic signal distrubance occurs, in which case there will be a relatively large, fast output signal change which may go positive or negative in direction due to ringing associated with e.g., noise impulse interaction with the RF circuitry of tuner 10. Thus, it may be desirable to include a second comparator circuit arranged with a threshold of opposite polarity in order to detect synchronous detector output signal excursions of either polarity. Still further, two comparators may be logically interconnected to produce an output signal only for a response known to be correlated to an impulse noise ringing characteristic.
The comparator output signal is conditioned by pulse generator 62 and applied via terminal 63 to enable/disable a switch 64 for affecting video signal substitution within comb filter 18. Pulse generator 62 may be a monostable multivibrator which generates a pulse of prescribed amplitude and duration (typically in the range of 1-10 microseconds). Test data indicates that impulse noise duration is generally grouped in the 1-3 microsecond range, and a three microsecond control pulse is sufficient to compensate for the majority of noise impulses. On the other hand, it may be desirable to terminate video substitution at a specified time each interval, for example, during the horizontal blanking period. In this case the pulse generator may be an edge triggered flip-flop, triggered on (set) by the comparator output signal and off (reset) by a separate timing pulse, e.g., the horizontal synchronization pulse.
Comb filter 18 is realized with charge transfer device technology and includes a charge coupled device (CCD) including a 1-H (one horizontal image line) delay line integral with CCD summing sections and sample and hold (S/H) circuitry to accomplish the requisite transversal filter function to comb the vido signal for separation of the interlaced luminance (Y) and chrominance (C) signals as is described in U.S. Pat. No. 4,158,209 issued June 12, 1979 in the name of P. A. Levine. The CCD delay line is actually a plurality of serially connected storage elements from which sampled data is successively translated from one element to the next element at a predetermined rate. The signal may be non-destructively tapped from any one or more of these storage sites in order to realize a particular signal delay. An adder 66 averages two samples of the video signal that are tapped one-half of a chrominance subcarrier cycle before and after a 1-H delay. The tapped and delayed signal is conditioned by adder 66 to have its chrominance signal shifted 180� (one-half of a chrominance subcarrier cycle) with respect to the untapped delayed signal remaining in the CCD device in order that the tapped and delayed signal will have the proper chrominance phase to serve as a substitution signal. A 1-H delayed signal is available at the output of adder 66 and is applied to terminal 69 via amplifier 68. The signal available at terminal 69 is then coupled to comb filter input terminal 70. For a more detailed description of the circuitry for phase inverting the chrominance signal see U.S. Pat. No. 4,272,785 issued June 9, 1981 to J. S. Fuhrer. By way of example, comb filter circuit 18 is available from Tokyo Shibaura Co., Ltd., Japan, as the T 3928 integrated circuit.
Real time video signal is applied to terminal I7 of the comb filter and then applied to input connection 71 of the CCD register via the switch 64. If signal defects, i.e., noise, have been determined to be present in the real time signal, the 1-H delayed signal from terminal 69 is temporarily substituted for the real time signal supplied from video detector 16 by positioning the switch 64 to access the 1-H delayed signal applied to comb filter input terminal 70. Thus, the signal previously delayed by one horizontal image line is recycled in the CCD register.
The foregoing demonstrates an advantageous technique of substituting delayed signal for real time signal in the video signal processing path of a color receiver utilizing a particular type of comb filter. An alternative arrangement for use in black and white TV receivers may comprise a simple 1-H delay line and a switch. In that case, the delay line has its input terminal connected for receiving a real time video signal and the switch selectively applies either the real time signal, or the 1-H delayed signal from the delay line directly to the further processing circuitry. In this type of arrangement, the video signal does not recycle in the delay line but merely pases through once subject to being delayed one image line. For a color signal, a delay corresponding to an integral number of horizontal line periods may also be used to effect the same basic result. For example, for an NTSC color signal, the delay circuit may comprise a 2-H delay line, in which case it is unnecessary to invert the phase of the chrominance component of the video signal in order that the chrominance component will have the proper phase to serve as the substitute signal.
Thus, what has been described is a system in which impulse noise or defect detection in the video signal is advantageously performed in conjunction with a PLL sound signal detector for detecting the audio information. While separate IF amplifiers 30 and 36 are used in the embodiment shown in the FIGURE, a common IF amplifier can be used for amplifying the IF picture and sound carrier signals provided at the output of buffer 24, such as described in copending U.S. patent application Ser. No. 412,898, now Pat. No. 4,470,070, entitled "FREQUENCY TRANSLATION PHASE-LOCKED LOOP SOUND DETECTION SYSTEM UTILIZING A SINGLE IF AMPLIFIER" filed Aug. 30, 1982 in the name of P. D. Griffis and assigned, like the present application, to RCA Corporation. Also, the NTSC frequencies mentioned herein are merely illustrative and can be appropriately scaled for operation at other frequencies in the NTSC, PAL or SECAM television systems.
Additionally, although PLL 22 is shown including mixer 34 for deriving the 41.25 MHz frequency translated signal for application to phase detector 38 and synchronous detector 54, the nominal frequency of the VCO output signal could be changed to the same frequency as the sound carrier frequency and be coupled directly to detectors 38 and 54, such as shown by the dashed line in the FIGURE, without being frequency translated by mixer 34. Of course, in this alternative embodiment, mixer 34 is omitted and would not be available to eliminate any common mode FM of the picture and sound carriers.
Furthermore, it should be understood that although in the described embodiment the defect control signal is used for controlling the substitution of delayed video signal, the defect control signal could be used in conjunction with other noise suppression or compensation circuitry. For example, the defect control signal could be used to render the automatic gain control system of the television receiver insensitive to noise impulses (impulse noise) in the video signal.
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Anybody know what's going on with the ISWA (International Submission wrestling association)? Are they still operating (i.e. events)?
Anybody know what's going on with the ISWA (International Submission wrestling association)? Are they still operating (i.e. events)?
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Townsend is a Wrestling/Grappling Coach at Xtreme Couture MMA in Las Vegas, Nev. working with Mixed Martial Arts legend Randy Couture, who was also a U.S. Greco-Roman wrestling World Team member.
He serves as Head Wrestling Coach at Marc Laimon?s Cobra Kai jiu-jitsu in Las Vegas, Nev. Cobra Kai jiu-jitsu placed two athletes in the finals of USA Wrestling Grappling World TeamTrials, runners-up Jeff Glover and Simpson Go. Glover will compete at the Grappling World Championships in Turkey.
?I am honored to be able to represent the USA as a coach of the first-ever U.S. Grappling World Team. It will be nice to use my knowledge of the rules to develop strategies, techniques and training methods necessary to defeat our foreign competition and bring home gold medals for the United States,? said Townsend.
Townsend serves as a member of the FILA World Grappling Committee, the group within international wrestling which manages Grappling worldwide. Within the committee, Townsend serves as Technical Director for coaches and officials education, as well as rules oversight. He is one of the authors of the FILA Grappling Rulebook.
He was the founder and President of the International Submission Wrestling Association, a Grappling organization that hosted competitions, as well as trained officials, coaches and athletes. With the ISWA, Townsend developed the rules of Grappling which have been adopted by FILA. The ISWA ceased operation after FILA and USA Wrestling adopted Grappling as a style of wrestling.
Townsend has been a key individual in putting together USA Wrestling?s existing Grappling program right from the start of organizational meetings this winter.
Townsend served as an assistant wrestling coach at Cal-State Fullerton from 2000-2005. He was a varsity wrestler for Syracuse Univ. and Hofstra Univ. Townsend was a two-time Maryland state high school champion, and was an All-American in freestyle at the Junior Nationals and Espoir Nationals.
?This is huge for the development of Grappling as a legitimate international amateur sport,? said Townsend. ?USA Wrestling has become a leader in the Grappling movement and a model for the implementation of this style into existing national wrestling federations around the world."
Thanks for the reply. It's good to hear Townsend got to move forward with his dream. I wrestled for him at Cal State Fullerton at 133lbs. Knew he was in Vegas, but lost track of him and the ISWA. It seems like everything is going well. Thanks again!
Can u PM me if you are in contact with him at all or just reply to this post. I want him to be involved in my site, and promoting his team members would be my privilege.
Last edited by MatDaddy; 06-09-2008 at 01:37 AM.
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