text:
- Research
- Open Access
Maternal smoking promotes chronic obstructive lung disease in the offspring as adults
- D Beyer1,
- H Mitfessel2 and
- A Gillissen1Email author
© I. Holzapfel Publishers 2009
- Published: 7 December 2009
In utero and/or childhood environmental tobacco smoke exposure is well known to adversely affect lung function and to depreciate child's health in many ways. Fewer studies have assessed the long-term effects on COPD development and disease severity in later adulthood.
COPD patients were interviewed using a structured questionnaire regarding their personal as well as the smoking habits of their parents. Data were compared with the disease history, e.g. COPD exacerbation rate, and their lung function data.
Between 2003 and 2004 COPD patients were recruited a) in a private practice specialized in pulmonary medicine (n = 133) and b) in a hospital (n = 158). 75% of their fathers and only 15.4 of all mothers smoked regularly. COPD patients from smoking mothers had lower FEV1 predicted than those raised in household without maternal smoking exposure: 39.4 ± 9.5% vs. 51.9 ± 6.0% (P = 0.037). Fathers had no effect on FEV1 regardless if they are smokers or non-smokers. Rate of severe exacerbations requiring hospitalization remained unaffected by parental second hand smoke exposure.
Maternal smoking negatively affects lung function of their offspring even in late adulthood when they develop COPD. It even aggravates the cumulative effect of active cigarette consumption. Clinical course of the COPD remained unaffected.
- chronic obstructive pulmonary disease, COPD
- second hand smoking
Second hand smoking is well known for its deteriorating effects in man. It increases significantly recurrent colds, cough and phlegm, throat problems, and the presence of respiratory symptoms by roughly a factor of 2 [1, 2]. It even has been linked to increase the risk for lung cancer by the factor of 1.3, particularly when the exposure occurs before the age of 25 [3]. Cigarette smoking is still quite common in many parts of the world. In parallel, exposure to second-hand smoke continues to be widespread despite declines in smoking in developed countries and despite evidence of serious health effects in infants and children. In addition to whatever effects direct and postnatal second-hand tobacco smoke exposure have on the occurrence of asthma, bronchitis, cardio-vascular diseases and impaired levels and growth of lung function in adolescents, there is an underlying alteration in the prenatal and early postnatal development of the structural and mechanical characteristics of the lung that contribute substantially to these deficits. These developmental effects are important contributors to the future risks for impaired pulmonary function in adolescence [4, 5].
While the detrimental effect of parental smoking exposure on the respiratory health of children is well known since years [6], information on the long-lasting consequence on human health, particularly regarding the development of diseases like chronic obstructive pulmonary disease (COPD) is sparse [7, 8]. Consequently, the purpose of this study was to assess the risk of environmental smoke exposure at home during childhood a) for COPD severity, b) for disease instability defined as higher exacerbation rate in later adulthood, and c) whether it might have an add-on effect to yearlong active smoke exposure.
Patients and Methods
This study has been approved by the ethics committee of the Saxonian Physician Association, Dresden/Germany (Code No.: EK-AMG-89/2003). Patients who refuse to participate or who were unable to understand the written informed consent form were excluded.
A structured 11 pages questionnaire was used to question prospectively COPD patients for their family and social history, previous occupation and possible occupational exposure status, parental and personal smoking habits. Diseases during childhood and adolescence were recorded as well as the year of COPD diagnosis and frequency of exacerbations, defined as those that required hospitalization and treatment with antibiotics and/or systemic corticosteroids [9]. At the time of the interview, following parameter were gathered from the patient: anthropometric data, dyspnoea using MRC-scale, lung function, blood gas analysis, and a current medication. To exclude concurrent diseases as defined below, patients were examined using EKG, echocardiography, X-ray of the chest.
Inclusion criteria were: COPD according to the GOLD (Global obstructive Lung Disease [10]) definition of all stages, age ≥ 30 years, negative reversibility testing (FEV1 < 12% and < 200 ml, 30 minutes after inhalation with salbutamol 400 μg). Patients were regarded non-eligible, when the following applied: severe concomitant co-morbidities in an acute deterioration or in chronic disease stage such as heart failure (NYHA ≥ 3), myocardial infarction ≤ 6 months, pneumonia and other acute infectious disease, asthma as defined by GINA (Global Initiative for Asthma [11]), interstitial and other lung disease except COPD, chronic kidney disease requiring dialysis, sleep apnoea syndrome, any malignant diseases, occupational disease of the lung (e.g. silicosis, asbestosis). For this bicentre trial COPD patients were recruited a) when admitted to the in-patient or out-patient unit of our centre (Robert-Koch-Hospital), an academic teaching hospital specialized in the management of acute and chronic pulmonary diseases including lung cancer, and b) in a private practice of a pulmonologist.
Statistical Evaluation
On the basis of earlier studies - whereas we choose paediatric studies due lack of comparable investigations with COPD patients - we calculated a sample size of 250 patients [12–14]. The software SPSS Version 11.5 was used for all statistical calculations. Unless otherwise stated data were compared using a t-test, and Mann Witney-U and the Chi2 tests. All efficacy analysis used the intent-to-treat population, defined as all included patients. The study was designed to show the influence of parental (father or mother) second hand exposure on the development of COPD. Within the predefined error probability, a significant influence was accepted at P ≤ 0.05.
Base data of patients.
Non-Exacerbaters (NE)
> 1 Exacerbation (1E)
Statistics/test used
Patient number
n = 133
n = 158
Age (y)*
67 ± 10
67 ± 10
P = 0.615/t-test
Gender (n, women/men)
P = 0.678/χ2-test
Age* at beginning of cigarette smoking
19 ± 5
20 ± 6
P = 0.652/t-test
Pack years
36.5 ± 20.6
37.9 ± 24.1
P = 0.371/t-test
Non-smoking since years (before interview)
13.2 ± 12.0
12.2 ± 9.9
P = 0.154/t-test
FEV1 % predicted*
62.7 + 20.9
9 43.1 + 20.9
9 P < 0.001/t-test
FEV1 (l)
1.6 ± 0.7
1.2 ± 0.61
P < 0.001/t-test
FVC (% predicted)
73.4 ± 18.6
59.7 ± 18.1
P < 0.001/t-test
FVC (l)
2.4 ± 0.9
2.2 + 0.7
P < 0.001/t-test
62.5 ± 15.1
56.6 ± 18.0
P < 0.01/t-test
Rt$ (cm H2O/l × sec.)
0.57 ± 0.33
1.19 ± 1.29
P < 0.01/t-test
Reversibility test§
8.1 ± 3.5
5.71 ± 4.6
P = 0.452/t-test
110.5 ± 47.9
129.5 ± 61.3
P = 0.768/t-test
pO2 (mmHg)
66,8 ± 13,0
58,8 ± 16,2
P < 0.01/t-test
pCO2 (mmHg)
37.5 ± 5.8
42.9 ± 12.7
P < 0.01/t-test
Patients (%) reporting chronic bronchitis (WHO definition)
P = 1.000/χ2-test
pneumonias or asthma (age* < 18 y)
Beginning of exercise induced dyspnea (age, y*)
49 ± 16
46 ± 17
P = 0.186/t-test
Reported exacerbations# (n, 2003-1980)
P < 0.001/U-test
In contrast to never-exacerbaters (NE), patients with ≥ 1 exacerbation (1E) reported that they had to be hospitalized about 3-times (mean) within the last 24 years because of severe deterioration of their disease. At the time of interview 58% of all patients were exsmokers (= non-smoking ≥ 1 year), 22% smoked actively and 19.2% claimed to be live long non-smoker.
COPD patients estimate smoking habits of their parents.
Smoker/non-smoker (%)
Cumulative cigarette exposure
(% positive answers)
< 10 py
10-20 py
> 20 py
Ca.10% of adult smokers demonstrate an increased decline of lung function with age [15, 16]. The effects of passive smoke exposure vary with genetic factors, gender, race and exposure to other pollutants [7, 8]. Exposure to environmental tobacco smoke during childhood and subsequent active smoking in later adulthood both further aggravate symptoms and have a negative effect on lung function, among others [17, 18].
This paper focuses on the effects of second-hand smoke in addition of direct exposure on the respiratory health of the lung function and frequency of exacerbations in COPD patients. Once birth cohort studies on this subject are unrealistic due to necessary long observation time, we choose an interview based approach in which COPD patients comment on their active cigarette smoking habits and second hand exposure during their childhood at home. Interestingly, maternal smoking resulted in further lung function decline in our patients, and thus has an additional effect to active smoking in later life. Although fathers smoked much more than theirs wives, paternal smoking was no contributor to the future risk for impaired pulmonary function. Based on earlier observations [19], we may speculate: Because mostly mothers but not their fathers raise children, the exposure to maternal cigarette smoke are much more intense than from paternal second hand smoke. Our patients could not know whether their mothers also smoked during pregnancy from obvious reasons. Assuming that this was the case in most instances, intra-uterine and inhalant second hand exposure may have contributed to lung function reduction as observed in our cohort [20]. Jurado et al [21] and others reported that maternal smoking increased the risk of wheezing with colds, cough with phlegm and asthma in the offspring up to 3-fold. The detrimental effect depended on the intensity of smoke exposure and if it occurs only prenatal, postnatal or both. Smoking fathers did not exert this effect or their influence was significantly smaller which was confirmed by our data [22–24].
Our results are very well in concurrence with published data indicating that direct and postnatal secondhand tobacco smoke exposure promotes the incidence of asthma and bronchitis, and impairs the growth of the lung [20, 25]. Tobacco smoke alters very early in life prenatal and early postnatal development of the structural and mechanical characteristics of the lung that contribute substantially to these deficits. These developmental effects are therefore important contributors to the future risks for impaired pulmonary function and COPD [26, 27].
Lower FEV1 in the cohort with smoking mothers did not correlate with increased rate of severe exacerbations requiring hospitalization. This amazes considering, that patients with FEV1 < 50% predicted has an 2.6-fold higher risk to become again hospitalized due to an COPD exacerbation [28]. Inversely reduces a FEV1 increase of 10% the exacerbation rate by 28% [29]. FEV1 values are by and large poor predictors for future exacerbations rates, because many patients with low FEV1 may be stable and patients with lower disease stages may frequently exacerbate [30, 31]. Good therapeutic management as demonstrated in studies using tiotropium bromide and inhaled corticosteroids or smoking cessation may help reduce exacerbations [9, 32–34]. All these factors could have diminished in our study a possible influence of environmental tobacco smoke exposure in childhood.
Our study has various disadvantages. Although the data were collected prospectively, we relayed predominantly on individual mental capacity for incidents dating back decades in the past. Objective methods to verify patient's strength to remember this kind of information is, to our knowledge, not on hand. We omitted therefore intelligence tests and related assessments. The study design did not allow the differentiation between in utero and post-natal smoke exposure. We only can assume that smoking mothers smoke also during pregnancy and not just after the delivery of their child. According to previous reports, about one third of smoking mothers stopped during pregnancy [35]. But after successful smoking cession during pregnancy approximately 10 - 60% of all women relapse to smoking during the next 6 months. 34% of all children are exposed to maternal smoking [35, 36]. Considering these data, one can reckon that over 70% of our patients reporting second hand smoke exposure during childhood were also exposed in utero.
We hereby demonstrate the reliability of data collection from COPD patients even if the event date back many decades, and we confirmed for the first time with this approach that maternal smoke exposure effects COPD severity, which even adds up to active smoking.
Conflicts of interest
The study was supported by Boehringer Ingelheim Pharma KG, Germany.
Authors' Affiliations
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You've probably heard about home management binders. Perhaps you've thought one wasn't for you. But I personally think a home management binder can potentially help anyone–and I think they can be incredibly helpful if the person who manages the home becomes incapacitated or unexpectedly passes away.
For many, a home management binder seems like one more piece of clutter, one more thing to keep up with. But handled appropriately, a home management binder can be very helpful in keeping you up to date on weekly, monthly, quarterly and yearly tasks that must be completed; keeping you focused with your home projects; limiting your grocery shopping trip to just one time per week; and maintaining enough food in your cabinets. So if you are one of those skeptics, I suggest you assemble a binder and try using it for a few months before brushing off the idea.
The Goal
You should create a binder that includes, at minimum, a calendar of important dates and appointments, a master to do list of tasks that need to be handled around your home, a menu plan, a shopping list, and a password log.
Suggested Supplies
- A three-ring binder, at least one inch thick
- Copy paper or cardstock for your binder's cover
- Cardstock for your binder's spine label and for dividers if you intend to make those yourself
- Index dividers if you don't intend to make dividers yourself
- Labels (I suggest Avery 5160 or similar generic labels.)
- Printer and printer ink or toner cartridges, depending on your printer, or other art supplies such as pens, markers, etc. if you intend to design your own cover, labels, and dividers
How to Assemble a Home Management Binder
- Obtain a three-ring binder. I use a one-inch binder and have never needed a larger one. Because I have multiple binders in my home, I do use a clear view binder so that I can put visible covers on it. I would suggest that you do the same, especially if you have students who have similar binders in your home. You can use a binder you might have lying around your house, such as one your child used last school year or you received at a continuing education class. You can also buy one. I recently saw them on sale at Dollar Tree.
- Create a cover for your binder. You want your home management binder to be noticeable and easily located, and a cover helps. You can create your own binder cover with just a pen–or let your kids design one. I have also included a free printable home management binder cover on my site. It is available exclusively to my newsletter subscribers. Click here to subscribe and receive access to it. If you do not have a clear view binder, you can skip this step or, better, simply slap an adhesive label on the front of your binder.
- Create a spine label for your clear view binder. A spine label makes your binder easier to locate on a bookshelf. I have a free printable one-inch binder spine for home management binders available on my site. Again, it is available exclusively for my newsletter subscribers. Click here to subscribe and receive access to it. I suggest that you print your spine label on cardstock rather than on copy paper to make inserting it into a clear view binder significantly easier. If you do not have a clear view binder, you can skip this step or, better, place an adhesive label on the spine of your binder.
- Buy or create at least four dividers for your binder. You can purchase index dividers if you have none. If you have a Silhouette Cameo and some 12×12 cardstock, I have a free Silhouette Studio index divider cut file available on my site to enable you to create your own dividers at home. The Silhouette Studio file is available exclusively for my newsletter subscribers. Click here to subscribe and receive access to the file.
- Label your dividers. For simplicity's sake, I suggest printing simple words on Avery 5160 labels if you have them. If you do have Avery 5160 labels, you can download a file on my site to make those labels in a hurry. But you have to be a newsletter subscriber to gain access to that file. So click here to subscribe and receive access to the file. If you're going to make your own labels, print or write labels for the following titles:
- Calendar / planner
- To do list
- Menu plan / shopping list
- Take-out menus (optional)
- Password log
- Address labels (optional)
Next Steps
Your next step will be to print and complete a calendar/planner, a menu plan and shopping list, a password log, and a to do list to insert inside your home management binder. I have files for all of these available as free resources on my blog. Click here to subscribe and receive access to the file. I'll discuss these in detail on the blog in the coming week.
How Did You Do?
Did you assemble a home management binder, or did you already have one? How long did this task take you to complete? Let me hear from you in the comments below!
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To make it fun [for me] I made a card [using one of the retiring sets; Exclamations], and I am also entering this into the Happy Little Stampers July Anything Goes with Dies Challenge.
Just a basic CAS card, but I love it! I will miss this set too, just sayin :).
Meanwhile, thanks HLS for a fun challenge - and to you all - check out that sale!
Thanks for stopping by~


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News Archive
Garcia Take Lead
August 8 2002
By Bryan Hersh of 49ers Paradise
THIS CONTENT IS COPYWRITED, REDISTRIBUTION OF IT (including copy/pasting it to a message board, forum or bbs) IS PROHIBITED AND COULD RESULT IN LEGAL ACTIONS - feel free to quote up to 1 paragraph providing a source link to http://www.49ersparadise.cjb.net is included
Earlier this off-season quarterback Jeff Garcia stated that he wanted to take on more of a leadership role this season. Actually, Garcia had already begun increasing his role with the team towards the end of last season, when coach Mariucci appointed him as a player who would be involved with the coaches on game planning. But this season, Garcia wants to be more vocal.
Veteran leadership if key to any sports franchise, and Garcia is finally in a position where he can really step up and lead without players questioning whether he has the right to be doing so. Garica has already stated that he wants to open communication with his receivers during the game to try and get a better understanding on how to attack the defenses they are faced with.
Lately though, Garcia has been speaking up, and trying to let the younger players know that they have to pick up there games. Garcia clearly recognizes the potential of the 49ers current squad, and wants the entire team to be on board and playing at their best when the season roles around.
"I realize the starters weren't in," Garcia said, "but that doesn't matter. This team is only as good as its depth. It's important to realize they're fighting when they're out there. They need to be stronger, and more willing to fight at the highest level.
Comments like these would have seemed out of place a year ago - when even Garcia himself had to be wondering if he could repeat his first Pro Bowl experience. But this season, there is no question, when Garcia says something, the team listens, and so having him speak out like this, telling the depth players exactly what needs to be done, is certainly a great step as he tries to increase his leadership on the team.


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Well I'm a wet person considering I swam 42,900 yards in August. That seems pretty much what I've settled down to swim per month.
Past Months To Compare -
In July I swam 29,550 yards.
June I swam 42,000 yards
May I swam 47,950 yards.
April I swam 44,950 yards.
March I swam 62,125 yards.
February I swam 41,750 yards.
January I swam 42,300 yards.
In 2011 I've swum 300,525 yards.
I looked back at older posts and the high 40's is what I use to train at. Of course I was swimming at faster intervals. Hopefully as the water and air temps fall I'll start to amp up my intervals. Actually in today's workout I felt like my body was adapting and I was ready to go faster. I've had a few false starts like this in the past few months so hopefully I can keep at it. I'll find out in October at my next meet.
As for my running in August I gotta say is pretty bad. After having hip issues in June and July I really was hoping to get back into running. I even tried to motivate myself into running by making of goal of running 16 miles in August. Well I didn't make it to 16. Blech! For the month I ran 11.03 miles. I will admit to a dumb accident which left my ankle and foot injured for a week. But compared to a year ago I have no base to start training a half or full marathon.
With that said I won't run the Disney World Marathon in 2012 or the half. Thinking that I'll continue to run and at the end of September have a 1/2 or full marathon training program. Maybe to a spring 2012 event. My goal for September will be to run 16.5 miles. Yeah, not a big goal but hopefully doable.
Compare with
At least I'm running and swimming. That is something in itself.


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Our idea of sound turned upside down with Thomas Edison's invention of the phonograph in 1877. Never before could we record a sound and reproduce it on demand. From that point on, we obtained more and more access to audio. Now we can turn on the radio in a car, at home, or online to play a song. We can buy the same song and keep it on an iPod in our pocket, along with thousands of other songs. We can even record ourselves singing the same song. The Vis Center's research goes beyond playing back sounds and works to manipulate audio input and output.
Audio research at the Vis Center delves into electronically focusing on the speech of a single individual in a noisy room, such as a cocktail party buzzing with chatter. The recording system uses a microphone array of up to 40 microphones scattered throughout a room to pick out individual voices. As the sound leaves a certain location, it hits the microphones at different times. We can use this time difference to find a sound source's location. Then, we use an algorithm to pick out the sounds we want to hear, and suppress noise and other voices. Typical microphone arrays require the microphones to be in a regular arrangement. Researchers at the Vis Center focus on using random arrangements of the microphones, making analysis of the sounds more difficult but making the system more applicable to real world situations. This technology is valuable for criminal justice and law enforcement agencies to covertly record and listen to conversations in noisy areas. The FBI does not have the luxury of placing microphones in a regular arrangement – they must hide the microphones wherever they can.
Not only do we conduct research on recording audio, but we also research audio output. We want to recreate the impression of a sound source where there is no loudspeaker. Speakers traditionally need to be in a regular arrangement, much like the microphones in the microphone arrays. However, this is not always possible. In immersive displays, there may be a 3D projection of a person, but you cannot put a loudspeaker there because the speaker would disrupt the image. Thus, we need to be able to scatter the loudspeakers irregularly in places where they will not disturb the projection, but also create a virtual sound source. The US army put this technology to use for combat simulation. To simulate sniper fire with no visual cues, classic technologies like a Dolby 5.1 array were not accurate enough. They needed our system of randomly distributing the speakers to create a virtual sound source.
Home entertainment and computer gaming will vastly improve with the implementation of our research. In classic home entertainment systems, the speakers must be regularly arranged to create a "sweet spot" for audio. However, some homes have architecturally constrained viewing spaces so that you cannot put the sofa in front of the screen with the loudspeakers. With our system of randomly arranged speakers, we can move the "sweet spot" anywhere in the room.
NOTICE: Some websites to which these materials provide links for the convenience of users are not managed by the University of Kentucky. The University does not review, control, or take responsibility for the contents of those sites. Equal Opportunity/Affirmative Action Employer.


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Does the classic Ford Bronco arouse fond memories? Your memories may be of the cool ride as well as the rough ride. The folks at Classic Ford Broncos take the vintage appeal of the early Bronco we all love and add the modern mechanicals that will make you love driving it. Classic Ford Broncos take older, worn out trucks and turn them into reborn works of art that are meant to be driven. There is no mass production line, just a small crew in a small shop.
If your dream is to drive a classic Bronco with today's mechanicals, this is where to begin. Stopping when you want to stop isn't a sign of weakness – it's a sign of modern Wilwood high-performance disc brakes. The crew blends their aesthetic requirements with your personality, and the result will be an envy-inducing Bronco that will stand out in any crowd. Just get in line. There are only 8 slots.
Gear For Life. Free for Life. Fill your boots with weekly updates.


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With easy out-of-the-box setup, integrated virus and malware protection and feature-enhancing updates, ASUS Chromebox starts up in seconds to get you to your favorite websites and apps instantly. And why a box? Because with a box it houses a processor powerful enough for ultrafast multitasking performance in Chrome OS and the connectivity options to choose your own screen size, connect to multiple displays at once and even attach to any VESA-mountable monitor or HD TV. All this starting at only $179, ASUS Chromebox is the always-new computer that just makes sense.
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It may be the world's smallest Chrome computer, but ASUS Chromebox is big on connectivity. It comes with high performance 802.11 b/g/n dual-band wireless, four USB 3.0 ports for fast data transfer, HDMI and DisplayPort for connecting to the latest displays and HDTVs, including support for dual displays and an SD card reader for easily accessing stored photos and documents.
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轉載請注明:友和 YOHO » Asus Chrombebox


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We have detected your current browser version is not the latest one. Xilinx.com uses the latest web technologies to bring you the best online experience possible. Please upgrade to a Xilinx.com supported browser:Chrome, Firefox, Internet Explorer 11, Safari. Thank you!
NGCodec's video encoding solution on AWS F1 Instances provides access to high quality video with low latency and low cost to more customers through the cloud. Leveraging Xilinx technology on the F1 Instance enables NGCodec to differentiate consumer experiences, reduce cost by 10x, decrease power compared to CPU-based solutions.


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YES! Is the new No in Yes Man
Person A: It's Friday lets go to the movies?
Person B: Nah… don't feel like it
Person A: Hey tomorrow the Knicks game is on, wanna watch it at my place?
Person B: Sorry, got some stuff I gotta do
Person A: I managed to gather our old friends for a poker night!
Person B: Damn, already got plans, sorry won't be able to make it
Do the above examples sound familiar to you? Then this movie might be just your thing, note with that though, you have to like Jim Carrey.
So you are a No-man right? What are you denying you aren't?! I am quite sure you are just fooling yourself right now. Within all of us there is a No-man and we love to give in to this person, just to be safe. You don't want to be put in situations you do not like, nor do you want to make decisions which could have a great impact, right? So we as humans tend to say no when things tend to get messy or generally irritating.
What you just read above is the basis of the movie. Jim Carrey plays Carl Allen a guy who never does a thing and after his breakup with his girlfriend says no to pretty much everything. His job is a loan officer who basically has to decide whether or not someone gets a loan. As a No-man, this job fits him perfectly!
Yet he is never content and that's where his friend Nick (John Michael Higgins) comes into play. He tells him about a Yes! seminar where you will learn to embrace life and say yes to possibilities you get in your life. It is here where he makes a pact with the yes-guru himself, Terence Bundley. So when he walks out of the seminar and a homeless guy asks him for a ride, to make use of his phone and to give him some money, he is saying yes. Of course his car wounds up with no gas, he has no money and his batteries are low the moment he wants to drive home again. It is then when he meets the lovely Allison, you know the chick from THE HAPPENING Zooey Deschanel. Because of this – and some more of these "coincidental scenes" – he starts to believe in this yes-therapy.
Things backfire when he does say no and you can imagine what happens when you have to say yes to a very old woman, who wants to give you a … well you know.
Throughout this movie he falls in love with Allison and she with him. She likes him because of his spontaneity and he thinks she is… wait I actually don't know why he likes her. Well she is definitely hot and I would not mind drinking a cup of coffee with her, so lets stick with the fact that she has a great personality. When she eventually finds out their whole relationship is based on him saying yes to everything, she starts questioning his sincerity. Carl now has to prove that he can break the yes pact and show Allison how he feels. This being a feel good movie you already know the ending will be a happy one. And that is why you went to see this movie in the first place. (And some of us just wanted to see the hot Zooey Deschanel.)
So to sum up YES MAN; the movie is a standard romantic comedy with a nice twist at end. But with the addition of someone who used to say no to life and start saying yes to possibilities, gives the audience a great feeling. I am quite sure that when the average moviegoer walks out of the theater he or she will ask whether or not it is a yes or no person. This makes the movie a better than average romantic comedy and worth at least one watch.


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Caldaro is a company with a gathered competence of more than 80 years within this niche of joysticks, pedals, sensors, handles and operators interface related products. We have been working on long term relations and business with our customers which have been leading to many successful cooperations over the years. We have been awarded OEM-supplier of the year on several occasions as well as other nominations in our customer's evaluation programs.
Our product offering is very niched but we serve many different customers in a number of different applications. Our products endure the harshest environments that suffer from extreme vibrations, changes of temperature, salt mist, water, oil, diesel and other fluids etc. From these applications and environments we have learned to design the products according to the customers wishes but also based on our know-how. We also have learned a lot regarding design for improved ergonomics, user friendliness, and how to maximize the output from the operator by making the handle as good as possible. Learn more about our Products and Tailored products by clicking on these headings.
The biggest area that still is to cover is the mobile hydraulics that are used with pilot valve controls today but will be changed to electronics controls (joysticks, foot pedals etc) in the future.
If you think about it there are many applications in which you will find a joystick, touch panel, foot pedal or similar type of Human – Machine Interface (HMI).


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|The Back Yard Show
September 12, 2005
Birgitte's first art exhibition recently took place in the backyard of her West Village apartment on September 12, 2005. Marcia Baczynski did a superb job curating the unconventional show. Roughly 40 people came to support Birgitte and the official first-viewing of her work.
|click image to advance
see all event pictures


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- Research article
- Open Access
Polarized secretion of Leukemia Inhibitory Factor
- Eric J Hill1 and
- Ann B Vernallis1Email author
© Hill and Vernallis; licensee BioMed Central Ltd. 2008
- Received: 16 July 2008
- Accepted: 18 September 2008
- Published: 18 September 2008
The direction of cytokine secretion from polarized cells determines the cytokine's cellular targets. Leukemia inhibitory factor (LIF) belongs to the interleukin-6 (IL-6) family of cytokines and signals through LIFR/gp130. Three factors which may regulate the direction of LIF secretion were studied: the site of stimulation, signal peptides, and expression levels. Stimulation with IL-1β is known to promote IL-6 secretion from the stimulated membrane (apical or basolateral) in the human intestinal epithelial cell line Caco-2. Since LIF is related to IL-6, LIF secretion was also tested in Caco-2 following IL-1β stimulation. Signal peptides may influence the trafficking of LIF. Two isoforms of murine LIF, LIF-M and LIF-D, encode different signal peptides which have been associated with different locations of the mature protein in fibroblasts. To determine the effect of the signal peptides on LIF secretion, secretion levels were compared in Madin-Darby canine kidney (MDCK) clones which expressed murine LIF-M or LIF-D or human LIF under the control of an inducible promoter. Low and high levels of LIF expression were also compared since saturation of the apical or basolateral route would reveal specific transporters for LIF.
When Caco-2 was grown on permeable supports, LIF was secreted constitutively with around 40% secreted into the apical chamber. Stimulation with IL-1β increased LIF production. After treating the apical surface with IL-1β, the percentage secreted apically remained similar to the untreated, whereas, when the cells were stimulated at the basolateral surface only 20% was secreted apically. In MDCK cells, an endogenous LIF-like protein was detected entirely in the apical compartment. The two mLIF isoforms showed no difference in their secretion patterns in MDCK. Interestingly, about 70% of murine and human LIF was secreted apically from MDCK over a 400-fold range of expression levels within clones and a 200,000-fold range across clones.
The site of stimulation affected the polarity of LIF secretion, while, signal peptides and expression levels did not. Exogenous LIF is transported in MDCK without readily saturated steps.
- Conditioned Medium
- Leukemia Inhibitory Factor
- MDCK Cell
- Apical Chamber
- Permeable Support
Most cells secrete a variety of cytokines, either continuously or in response to stimulation. The spatial regulation of cytokine secretion is important since it determines which neighbouring cells respond, but little is known about how cytokine secretion in the constitutive pathway is regulated. With their prominent apical and basolateral domains, epithelial cells have been studied extensively for their ability to sort membrane proteins to specific domains. Sorting depends on hierarchical signals and occurs in multiple stages of the secretory pathway [as reviewed in [1–3]]. Lipid raft-associated and independent pathways have been identified for transport from the trans-Golgi to the apical membrane [4]. Less is known about the sorting of secreted proteins. N-glycans and O-glycans as well as proteinaceous patches have been proposed as sorting signals. Lectins may cooperate with lipid rafts in the transport routes [5].
LIF is a secreted, glycosylated cytokine, of the IL-6 family, that exhibits pleiotropic activities in a wide range of tissues and cell types. LIF is produced by epithelial cells during development and during infection and inflammation, including in the intestine [6], the uterus [7] the lung [8], and the kidney [9].
When cultured on permeable supports, Caco-2 cells display the features of differentiated small intestinal enterocytes, forming polarized monolayers with tight junctions. We chose to characterise LIF secretion by Caco-2, since Moon et al. [10] made an intriguing observation about IL-6 secretion in these cells. They reported that IL-6 secretion was higher from the membrane domain (apical or basolateral) that was stimulated with IL-1β than from the non-stimulated domain, suggesting that secretion could be locally facilitated by a pro-inflammatory stimulus. Whether or not LIF is secreted in a similar way is interesting since LIF and IL-6 are related cytokines and may be co-regulated.
We also studied LIF secretion in MDCK, a renal cell line which displays characteristics of distal tubule cells and collecting duct cells [11]. MDCK were selected since the cell line is well suited to exogenous expression. Murine LIF (mLIF) and human LIF (hLIF) are easily distinguished from endogenous canine LIF in ELISAs. Inducible expression systems based on tetracycline-regulated promoters are available for MDCK which allow the effects of different expression levels on secretion to be measured. Few studies have systematically examined the effects of varying expression levels. Marmorstein et al. [12] conducted a study of vascular endothelial growth factor (VEGF165) and transforming growth factor β1(TGF-β1) secretion at different expression levels in a retinal pigment epithelial cell line (RPE-J) by varying the amount of adenovirus used to transfect the cells. Their results indicated that these cytokines were secreted in apical pathways that were readily saturated, suggesting specific transporters. For example, VEGF dropped dramatically from 100% apical secretion to 30% apical with increased expression levels. We tested whether this result could be generalized to mLIF and hLIF in MDCK.
Exogenous expression also facilitated study of the contribution of LIF signal peptides to LIF trafficking. Two secreted isoforms of LIF are derived from the murine LIF gene. The isoforms, corresponding to transcripts LIF-D and LIF-M, encode different signal peptides, which in at least some fibroblasts appeared to affect the localization of LIF protein. After cleavage of the signal peptides, the mature murine proteins had the same polypeptide sequence but the LIF-D protein was secreted and freely diffusible; whereas the LIF-M was secreted but remained associated with the extracellular matrix [ECM; [13]]. LIF-D and LIF-M transcripts are also produced from the hLIF gene [14, 15]; however, differences in localization of the products have not been reported. If, as in fibroblasts, LIF-M was associated with the ECM in epithelial cells, LIF-M secretion might be preferentially directed toward the basolateral membrane, where the ECM is present. To test this hypothesis, LIF-M and LIF-D were exogenously expressed in MDCK.
Our results suggest that exogenous LIF is predominantly secreted in an apical direction in MDCK, regardless of signal peptide or expression level, indicating a lack of specific transporters for LIF.
Caco-2 secreted LIF
The polarized secretion of a number of cytokines including IL-6 has been described in Caco-2 cells [10]. Secretion of LIF by Caco-2 has not been previously reported. Caco-2 cells were grown as a confluent monolayer in tissue culture flasks for two weeks to allow them to fully differentiate. Conditioned media was then collected over a 48 hour period and assayed for LIF-like activity using a BA/F3-mLIFR-mgp130 assay (hLIF will activate mLIFR/mgp130). The conditioned media displayed a LIF-like activity, which was detectable over background in up to a 1:16 dilution and did not support BA/F3 parental cells (lacking mLIFR/mgp130). A polyclonal antibody raised against eukaryotic human LIF inhibited the proliferative responses by 75% (data not shown). The neutralization by anti-LIF antibodies demonstrates that a large proportion if not all of the activity in the conditioned media is due to hLIF itself and not to other LIFR-dependent ligands.
An hLIF ELISA, which uses antibodies raised against glycosylated hLIF, detected 32 pg/ml hLIF in the Caco-2 conditioned media. This concentration is similar to what was predicted from the BA/F3-mLIFR/mgp130 assays. The ELISA result demonstrates that Caco-2 constitutively secrete small amounts of LIF.
Caco-2 preferentially secreted LIF from the basolateral membrane
Secretion of LIF and IL-6 by Caco-2 was influenced by the site of stimulation with IL-1β
Apical or basal treatment alone with IL-1β resulted in an increase in the total amount of LIF secreted, but the increase was not statistically significant. Simultaneous treatment of both sides (dual stimulation) resulted in a significant 7-fold increase relative to unstimulated cells (Fig. 1a). Differences between apical and basolateral stimulation were apparent when considering the polarity of secretion. With apical stimulation, 42% of the LIF was secreted apically, whereas with basolateral stimulation only 20% of the LIF was secreted apically, demonstrating that the site of stimulation is important in determining the direction of secretion (Fig 1b). Dual stimulation yielded an intermediate percentage secreted apically. The effect of IL-1β on LIF trafficking is not due to a disruption of the monolayer integrity since stimulation with IL-1β did not have a significant effect on monolayer permeability (data not shown).
The samples collected for LIF secretion were also assayed for IL-6 using an IL-6 ELISA. In contrast to LIF, IL-6 secretion was not detected in untreated cells (Fig. 1c). Apical, basal or dual stimulation with IL-1β all increased IL-6 secretion, but only the basal and dual increases were significant, reaching an average of 284 pg with dual stimulation. When stimulated at the apical membrane, 57% of the IL-6 was secreted into the apical chamber (Fig. 1d). In contrast, basolateral treatment with IL-1β resulted in only 33% secreted apically. With stimulation of both sides of the monolayer, an intermediate percentage was secreted apically. As with LIF, therefore, the polarity of IL-6 secretion was differentially regulated by IL-1β at the apical and basolateral membranes. The two cytokines show a closely related but not identical pattern of secretion.
MDCK secreted a LIF-like factor from the apical membranes
MDCK mLIF clones secrete LIF primarily from the apical membrane
Since the level of expression has been shown to influence the direction of secretion of growth factors, several stable MDCK Tet-off LIF cell lines were generated for study using the PBI-L expression vector. PBI-L contains a bi-directional promoter, allowing clones to be selected according to their expression of luciferase and/or LIF. To test the polarity of secretion at the minimum and maximum levels of LIF expression, clones were grown on permeable supports and analyzed both in the fully switched on state (no doxycycline; No Dox) or fully switched off (1 μg/ml Dox). Both mLIF and hLIF-expressing clones were generated since differences in their surface-exposed amino acids might result in differences in protein sorting.
As MDCK cells are responsive to stimulation with hLIF [17], the effects of mLIF and hLIF on monolayer integrity were first tested. Eukaryotic mLIF or hLIF (1 ng/ml) was added to both sides of confluent monolayers and the effects on TEER were tested after 24 hours. MDCK cells produced TEERs in the region of 700–850 Ω.cm2 after five days growth on filters. No significant difference in TEER was observed following stimulation with either mLIF or hLIF in comparison to untreated cells. At most small amounts of LIF were transported across the monolayer. After adding LIF to the apical or basal side, less than 5% of the recovered LIF was detected in the opposite chamber.
In order to identify any differences in the pattern of secretion between the LIF-D and LIF-M signal peptides, two MDCK Tet-off-mLIF-M clones were also examined over a range of expression levels (Fig 3c–f). Without Dox, mLIF-M clone F produced 11 ng of mLIF (78% in the apical chamber), whereas with Dox this clone produced significantly less mLIF (less than 1 ng) and secreted less of it apically (71%). Without Dox, mLIF-M clone S produced 8 ng of mLIF (66% in the apical chamber), but with 1 μg/ml Dox it did not produce detectable amounts of mLIF. The experiments were repeated with less Dox (Fig. 3e and 3f). In these experiments, clone S secreted without Dox, 31 ng of mLIF (70% in the apical chamber). With 1 ng/ml Dox, clone S secreted significantly less mLIF (253 pg), but secreted a similar percentage apically (74%). Thus of the two clones one showed a difference in the proportion secreted apically at different levels of expression and one did not. Both clones secreted the majority of the mLIF in the apical direction, which is similar to the pattern seen with clones expressing mLIF-D. There is no evidence for a difference between the two signal peptides in directing the polarity of mLIF secretion.
MDCK hLIF clones also preferentially secrete LIF from the apical membrane
The expression of pro-inflammatory cytokines and their receptors by normal (unstimulated) intestinal epithelial cells points towards a role for these cytokines in the normal growth and maintenance of the gut epithelium. The secretion of LIF by unstimulated Caco-2 cells is consistent with the detection of LIF from human colonic epithelial cells [6] and the presence of LIF in mRNA obtained from murine intestinal tissue [18]. Guimbaud et al [19] reported that Caco-2 cells did not make LIF perhaps because they used a less sensitive ELISA. Whilst the human colonic epithelium has been reported to lack LIFR expression, pericryptal fibroblasts express the receptor, suggesting an interaction of LIF produced by colonic epithelial cells with pericryptal fibroblasts [6]. LIF levels are increased in inflammation, as demonstrated by high levels of LIF in patients with ulcerative colitis [19]. In contrast to LIF, we did not observe constitutive secretion of IL-6 from Caco-2. This may reflect different physiological roles for LIF and IL-6 in the normal intestinal epithelium.
It is striking that the site of LIF and IL-6 secretion is influenced by the site of stimulation. Such preferential secretion displays a variety of features. It is observed with some cytokines and acute phase proteins but not all secreted proteins [10, 20], with only some inducers [21], and in a variety of polarized cell types [21–24]. The mechanisms which underlie preferential secretion at the site of stimulation have not been defined and merit future study. Enhanced secretion at the site of stimulation could result from a global response such as an increase in the amount of membrane incorporated into the stimulated membrane or a change in membrane composition. If the mechanisms are global, however, there must remain transport routes which are not affected by these changes. For example, Moon et al. demonstrated in Caco-2 that although IL-6 displayed a marked preference for secretion through the membrane stimulated by IL-1β, the secretion of complement component C3 displayed strong basolateral secretion regardless of the membrane stimulated [10].
Conditioned media from MDCK cells demonstrated LIF-like activity in BA/F3 mLIFR-mgp130. LIF expression by MDCK would not be surprising since collecting duct cells are known in rat to express LIFR and LIF. Following ischemia-reperfusion injury LIF is expressed more widely by the epithelial cells of the kidney, where it may participate in regeneration by stimulating the proliferation of epithelial cells [25, 26]. LIFR is expressed at both the apical and basolateral membranes of MDCK [17]. The apparent selectivity for apical secretion of a LIF-like activity observed here may reflect paracrine signalling between epithelial cells and is broadly consistent with the apical secretion observed in the exogenous LIF-expressing clones. A sensitive ELISA for canine LIF is required for a more definitive comparison between endogenous and exogenous expression.
We generated cell clones from MDCK Tet-off cells in which LIF expression was regulated by a tetracycline-inducible promoter. The clones demonstrated a wide range of LIF expression levels when the promoters were induced or repressed. Exogenous mLIF was mostly secreted in an apical direction irrespective of whether the signal peptide was from LIF-D or LIF-M. This suggests that the LIF-M signal peptide does not direct the secretion of LIF basolaterally toward the ECM in MDCK. We have not measured the binding capacity of the ECM for LIF in MDCK cells. If the binding capacity is similar to that observed in the osteosarcoma cell line UMR-106 [27], however, it would be easily saturated when LIF is highly expressed in the absence of doxycycline and is unlikely to have affected the measurement of polarity. Cell-type differences may be important in the recognition of sorting signals. When proteins that are polarized in epithelial cells are expressed in fibroblasts, they display specific secretion routes [28, 29], however, it is not known if routes used by endogenous proteins in fibroblasts are present in epithelial cells. LIF may also require a specific "sorting escort" as has been observed for components of secretory granules [30].
Interestingly, the level of LIF expression did not influence the proportion of LIF secreted apically and basolaterally. The high levels of LIF produced were comparable to those observed with VEGF when its expression level was manipulated in RPE-J cells [12], but there was no evidence in MDCK of a shift in LIF polarity with increasing expression comparable to the shift observed for VEGF (100% to 30% apical). This failure to observe saturation may reflect a lack of specific sorting receptors for LIF.
The polarity observed with exogenous mLIF and hLIF (70% apical) in MDCK may simply reflect the total volume of vesicles delivered to the apical and basolateral surfaces. This hypothesis is attractive since passive incorporation of LIF into vesicles would account for the lack of saturation. To our knowledge total vesicular traffic has not been measured in MDCK so it is not known what percentage secreted apically would be consistent with passive transport. Comparisons with other four-helical cytokines, however, suggest that intrinsic features of LIF are likely to play a role even in passive transport. Rat growth hormone which is unglycosylated displayed only 35–40% secreted apically when expressed exogenously in MDCK [31, 32], indicating differential sorting of LIF and growth hormone. LIF more closely resembles erythropoietin in its secretion pattern. Wild-type erythropoietin displayed predominantly apical secretion when exogenously expressed in MDCK. The percentage apical secretion dropped to about 50% after the removal of one of the N-glycosylation sites [33], suggesting that either the N-glycosylation serves as a sorting signal or it is required for the conformation of a proteinaceous signal. Further experiments could reveal if the glycosylation of LIF was similarly required for apical secretion. One way in which the glycosylation/conformation of LIF could be important is if it mediates a preferential affinity for the lipids that compose apical vesicles or tubules. Constitutive secretory proteins have been shown to be associated with the lumenal face of membranes in the secretory pathway [34]. If LIF can directly interact with lipid domains, it may explain why saturation was not detected for LIF. The high expression levels achieved with exogenous LIF will make experiments with confocal microscopy and detergent solubilization feasible.
The apical secretion of exogenous LIF and an endogenous LIF-like activity in MDCK contrasts with the basolateral preference seen in Caco-2. LIF is not unusual in this regard. Differences in secretion patterns of transmembrane proteins between these two cell lines have been reported and attributed to differences in the recognition of basolateral trafficking signals [35]. We haven't determined whether the preferential secretion of LIF at the site of stimulation is unique to Caco-2 or shared with MDCK. Similarly we haven't determined whether the lack of a role for LIF signal peptides and the absence of saturation are unique to MDCK. For each of these findings, further experiments are required to test whether they extend to both epithelial cell lines.
The results presented provide evidence for the secretion of LIF by intestinal epithelial cells and a LIF-like activity from kidney epithelial cells. Interesting questions have emerged as to how the site of stimulation is able to influence the pattern of protein secretion and how the secretion of exogenous LIF is directed to the apical and basolateral poles in the apparent absence of specific protein sorters. More mechanistic approaches are now required to pursue these questions.
Cell culture
Caco-2 cells were maintained in DMEM, high glucose, with L-glutamine (Gibco), 20% FCS (Labtech), 1% MEM (Gibco), 100 U/ml penicillin, and 100 μg/ml streptomycin (Gibco). BA/F3 mLIR-mgp130 cells were maintained in RPMI 1640 (Gibco), supplemented with 10% FCS, 50 U/ml penicillin, 50 μg/ml streptomycin, and recombinant human LIF (20 ng/ml). Parental BA/F3 cells were maintained in RPMI 1640 (Gibco), supplemented with 10% FCS, 50 U/ml penicillin, 50 μg/ml streptomycin, and 100 pg/ml IL-3 (R & D systems). MDCK Tet-off cells (BD Biosciences) were maintained in DMEM high glucose containing 50 U/ml penicillin, and 50 μg/ml streptomycin and 1 μg/ml puromycin (Sigma). Polarized secretion experiments were performed with cells grown on 4.7 cm2, 0.4 μm pore size polycarbonate membrane Transwell® inserts (Costar). Caco-2 cells were seeded into Transwell® inserts at a density of 4 × 105 cells/well. Media in each chamber was replaced every two-three days for fourteen days, after which time monolayer integrity was demonstrated by measuring transepithelial resistance (TEER; described below). For stimulation experiments, monolayers were stimulated with 1 ng/ml IL-1β (R & D systems) for 24 hours. MDCK cells were seeded into Transwell® inserts at a density of 106 cells/well. Media in each chamber was replaced every 2–3 days for five days, after which time monolayer integrity was verified by measuring TEER.
Epithelial monolayer integrity
The TEER of each monolayer was measured with an epithelial voltmeter using 'chopstick electrodes' (EVOM World Precision Instruments, USA). The mean TEER was calculated from three different positions. The intrinsic resistance of the insert (permeable support alone) was subtracted from the total resistance (cell monolayer and permeable support) to calculate the resistance of the monolayer. The resistance was corrected for surface area of the permeable support (4.7 cm2) and the TEER was expressed as Ohms cm2 (Ω.cm2). TEER was determined before the start and at the end of each period of collecting conditioned media.
Proliferation assay
Activity of conditioned media was assessed by a cell proliferation assay. BA/F3 mLIR-mgp130 is a pro-B-cell line that has been stably transfected with cDNA, encoding both components of the mLIF receptor: mLIFR and mgp130. The assay was performed as described previously [36]. Proliferation was measured using 1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT; [37]). Neutralization of LIF-like activity was achieved by adding 10 μg/ml of rabbit anti-hLIF (Chemicon; AB1886) to the assay.
LIF and IL-6 concentrations in cell supernatants were determined by specific ELISA: hLIF, hIL-6 (Bender MedSystems), and mLIF (Quantikine ELISA; R & D Systems). In our experiments, the detection limits for eukaryotic hLIF and hIL-6 were 10 pg/ml. The Quantikine mLIF kit detected bacterial mLIF down to 20 pg/ml. The kit also detected purified glycosylated mLIF that we prepared in eukaryotic cells.
Construction of LIF expression plasmids
mLIF cDNA encoding the LIF-D signal peptide was isolated from the plasmid pXMT2 mLIF-D [13] using the enzyme EcoR1. The fragment was subcloned into the plasmid PcDNA3.1- (Invitrogen) to create the plasmid PcDNA3-mLIF-D. Murine LIF cDNA encoding the LIF-M signal peptide was isolated from the plasmid pXMT2 mLIF-M [13] using EcoR1 and Xho1. The fragment was subcloned into the plasmid, PcDNA3.1 (-), to create the plasmid PcDNA3.1-mLIF-M. To ensure a comparable 5' end to the mLIF clones, hLIF-D cDNA was generated from a pre-existing PcDNA3.1-hLIF plasmid by PCR. The 5' oligonucleotide was directed to the 5' signal peptide and included a BamH1 site and the 3' oligonucleotide was directed to the 3' end of the LIF coding sequence and included an EcoR1 restriction site. The resulting fragment was subsequently ligated into PcDNA3.1 to create the plasmid PcDNA3.1-hLIF-D. cDNAs encoding the mLIF-D, mLIF-M and hLIF-D signal peptides were isolated from their respective PCDNA3.1 plasmids using the restriction enzymes Nhe1 and HindIII and were subsequently ligated into the plasmid pBI-L (BD Biosciences). All constructs generated by PCR were verified by automated DNA sequencing (Functional Genomics, University of Birmingham).
Stable expression in MDCK Tet-OFF cells
MDCK Tet-Off cells (BD Biosciences) were transfected with PBI-L-mLIF-D, PBI-L-mLIF-M, and PBI-L hLIF-D using calcium phosphate. Stably transfected cells were selected in media containing 100 ng/ml hygromycin and 1 μg/ml doxycycline and cloned using cloning cylinders (Sigma). Cells expressing exogenous LIF were identified by luciferase activity transcribed from the bi-directional promoter of PBI-L and by LIF ELISA.
Data Analysis
Results are presented as mean ± standard error of the mean (SEM) of three separate experiments in which duplicate wells were analyzed except where otherwise specified. For comparing total secretion in the presence and absence of doxycycline, unpaired t tests were performed. For comparing proportions, the data was first normalized by taking the arcsine of the square root of the proportion. When more than two results were compared, an analysis of variance (ANOVA) followed by the Tukeys test was performed.
Prof. J.K. Heath (University of Birmingham, UK) kindly provided the plasmids, PXMT2-mLIFM and PXMT2-mLIFD. Dr. Hill was funded by a PhD studentship provided by the School of Life and Health Sciences, Aston University. Preliminary work was funded by a Royal Society Research Grant to Dr. Vernallis. Neither funding source has influenced: the study design, the collection, the analysis and the interpretation of data; the writing of the manuscript; or the decision to submit the manuscript for publication.
Authors' Affiliations
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Service & Philanthropy
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Each year, Greek students raise and contribute tens of thousands of dollars and service hours to worthy causes. Students who enjoy supporting worthy causes, organizing campaigns and participating in events to help others will fit well in the our community.
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Hello List,
I am currently researing masking models - frequency spreading functions and time domain spreading functions (forward and backward masking).
I am looking for any new publications etc containing new masking models. These would be publications in the last 5 years or so that have introduced new masking models or new spreading functions.
I have been looking at the Frequency Spreading and Time Domain Spreading functions used in PESQ and PEAQ but I would like to find new models introduced since PESQ and PEAQ were standardized. In other words I want new masking models that have been developed in the past 5 years.
Also, I would be interested in any summary papers or comparison studies that have covered a number of speech or audio masking models.
I would be most grateful if anyone could help me out with information.
Many thanks.
Best Wishes,


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During my quest for a simpler cloth diapering routine I found Imagine Baby cotton prefold diapers. I had tried the Osocozy brand and was not impressed with the fit. They were really long and narrow and that made them hard to fit on a small, squirmy bottom. But the Imagine Baby prefolds were so soft and had a better width and more manageable length to them. I became an insta-fan.
The thing for me about prefold diapers though was when I folded them I ended up with a very bulky section in the groin. There had to be a better way!
Insert my Customized Prefold Cloth Makeover.
I began with my washed and dried prefolds. Mine had been used several times before so they didn't need prewashed but if you are starting with brand new prefolds you need to follow washing instructions before you begin.
Laying my prefold out flat, I used my rotary cutter to cut into the prefold toward the center where the absorbency is but not all the way to the stitching. I measured 3 inches down from the serged edges and then drove the cutter in at a very slight rounded angle.
*I apologize for the horribly focused photo. My camera is a little fickle*
Make this exact cut 3 more times so you have 2 on each long side. These will become flaps that you can then customize your fold and fit with.
Next I stitched along each of the cuts. You make choose your stitch based on your machine and skill. You can do an overcast stitch, a zigzag stitch, or a straight stitch. My first round I tried a zigzag stitch and after several washes seemed to have missed fabric in places and had lots of fraying. This time I just did a straight stitch.
It is simply dropping your needle and foot at one end and stitching in to the corner of your cut. I back-stitched at the beginning, the corner, and the end of each cut. Do this on all four cuts and there you go! Super easy!
Now, for the folding…
You can tri-fold the flaps, just as if you would do it before but without the added bulk.
You may also use the jelly-roll technique for added protection from blowouts!
You can also still adjust the rise just by folding the front down (shown above).
By doing this to my diapers I keep all the function and absorbency that I had before but with much less bulk. It also keeps our diaper changes quick and easier to do one-handed.
Here is my stash! It makes for easy laundering and packing a diaper bag!
Do you use prefold diapers? Would you try this with your diapers? Leave a comment!


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New interesting plot details on Batman v Superman have surfaced online – this time it's about the film's ending and Lex Luthor's plan.
Warning: Potential Spoilers Below
By now, fans would have assumed that Kryptonite will be of immense significance in the film for Luthor and Bruce Wayne/Batman (even though their intentions behind using it may be quite different).
An anonymous extra, who claimed to have worked on the Detroit set of Batman v Superman, revealed some key plot details to Geek Tyrant.
According to the extra, the ending of the film has already been filmed. One of the scenes shot featured Batman breaking into Lexcorp to steal kryptonite, and apparently Lexcorp had somehow gotten hold of General Zod's body.
He went on to talk about the timeframe of the movie and where it picks up, saying that it starts "as the battle between Superman and General Zod ends or is ending (which would explain all the damaged buildings and vehicles we've been seeing in photos) and Batman/Bruce Wayne is attempting to make his way to the Wayne tower in Metropolis."
Badass Digest confirms this big plot reveal and also includes additional details:
"Lex has been gathering pieces and debris from the World Engine and he has used them to synthesise Kryptonite. Meanwhile he has also gotten ahold of Zod's corpse, and it is the key to his master plan to defeat Superman. He uses that tissue to... well, that would be spoiling. But Superman hasn't been defeated many times in the past. You can probably connect the dots from here," states the report.
Meanwhile, Ben Affleck, in his latest interview with Entertainment Weekly, spoke about playing Batman.
"I'm right in the middle of shooting now. It's a process that I'll probably have better perspective on when I'm done. I can tell you that every time I do a role, it's the responsibility of an actor to get their physical self as close to the role as possible. And Batman's obviously got a set of expectations that are tough. So I spent a lot of time working out.
"Before I took the role, Warner Bros. gave me a bunch of past reactions to casting and said, 'Are you sure you want to get into this? This is part and parcel of these movies now. There's a lot of active fans with a lot of opinions.' To me, having been through a certain amount of that, it doesn't really... Everyone's entitled to their opinion. I wouldn't have taken the part if I didn't trust my instincts in terms of the filmmaking. I think Chris Terrio wrote a terrific script. Zack's a great visual director. And there's an interesting take. I wouldn't have done it if I didn't think I could do it," Affleck was quoted as saying.


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..and by breasts I mean weird pectorals.
WTF did she do?!?! Nooooo. Even Winehouse's set look better. Although the upside is, I'm sure Keith likes it...as in the pectorals, if you know what I mean.
Sunday, November 15, 2009
Ummm...Kidman's breasts are scaring me
Tags: breast implants, nicole kidman
1 comment:
- Canvas Photos said...
Yeah, these breasts are killin me too. But What's up with all of these celebs that don't know how to contain their breasts? Katy Perry?
- 5:01 am
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Pocono Manor Resort and Spa is located on 3,000 lush acres with ballrooms and ceremony sites overlooking the beautiful Pocono Mountains and comfortably accommodates groups of 25 to 300 people.
With more than 100 years of service, you can be assured that we will tend to every detail and provide you and your guests with exceptional hospitality and memories that will last a lifetime.
Adventure beckons at Pocono Manor Resort & Spa


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If you tuned into my last column, you will have probably witnessed me have right old dig at the scientific mainstream for choosing a mechanical and lifeless model of the universe with which to indoctrinate the "civilised" world. Fair enough it may sound a damn sight better than the picture painted by western religion, where one step out of line will have you condemned to fire and brimstone for eternity, but I find both to be wholly ineffective at describing the world as I see it.
The source of this great divide between science and religion is usually attributed to the philosopher Rene Descartes, who in the early 1600's pioneered the idea that the world of matter is purely mechanical; that atoms are just lifeless balls of energy which coast around the universe governed by simple physical laws. Although he and his contemporaries still believed in God, they had created a secular science by extracting God out of the physical world; the universe became a clockwork device, where God was the watchmaker and had no further involvement after winding it up. The roman catholic church accepted this, and the scientific revolution began - people could explore the physical world with a renewed sense of freedom as there was no longer any fear of committing heresy and getting burned at the stake.
One of the consequences of this, was that man was essentially promoted to a God-like status within the physical world. Even animals were believed by the intelligentsia to have no spirit in them, but were viewed as mere complex clockwork phenomena resulting from the laws of motion, and thus equally amenable to scientific experimentation. One source writes "They administered beatings to dogs with perfect indifference, and made fun of those who pitied the creatures as if they felt pain … They nailed poor animals up on boards by their four paws to vivisect them and watch the blood circulate, which was a great subject of conversation".
So the so-called "Enlightenment" got off to a pretty unenlightened start, and although we have come a long way since that particular kind of vile materialism, the belief that Earth is merely a resource to feed man's whimsical desire has persisted. It fueled the industrial revolution and underlies our entire world economy. Nothing is sacred except human life, and even then some of us are more sacred than others. In theoretical physics, the idea of a spiritually devoid universe has been explored to obscene levels. The creators of the universe, they say, are lifeless energy membranes floating in some hyper-dimensional vacuum and colliding with each other to produce "big bangs". Most of the collisions lead to "failed universes" and we happen to live in one which succeeded. But don't get too excited, because according to one recent article on BBC News, it's all going to end in cold dark emptiness, and there's nothing we can do about it.
You see, in between the big bang and the cold dark emptiness, the debris from the explosion accidentally happened to organise itself into what we call life. It's a total fluke, an uncountable series of highly improbable events, one after another. Our consciousness is just the latest in the chain; an illusion at the leading edge of the big bang explosion. Eventually it will all decay into absolute nothingness, just like before the accident. Now to be fair, it's probably one of the least ridiculous theories of the universe you can create without incorporating a spiritual dimension. But as many people will attest, this 400 year old paradigm of secular materialism is crumbling, and the global economy which is built upon it is sinking like the proverbial house on sand.
READ THE FULL ARTICLE IN LSD MAGAZINE ISSUE 6 (Below)


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Regarding the purchase of catalogs
From Sandnes Yarn's terms of purchase: "We only sell our catalogues together with yarns for a garment in the booklet, minimum 4 yarnballs per catalog. We reserve the right to cancel orders that do not meet these requirements."
From Viking Yarn's website: "When buying a catalog, you must also order a minimum of 4 yarns per catalog as well"
Drops Design has no problem to sell catalogues without yarn.
We follow these purchase terms: You cannot buy Sandnes Garn and Viking Garn catalogs without having at least 4 yarnballs in your shopping cart.
Product Review Guidelines:
What should a product review contain?
Your own experience focusing on the product in question: You must have purchased the item to write a review.
Please do not include:
- Experiences that are not about the product, such as experience in connection with support or return of the product in question.
- This is not a forum, so do not start discussion or reply to others who have written a review.
- Links, prices, availability or other time-dependent information.
- References to competitors.
- Shocking/improper word usage.
"Outpost butikk" reserves the right to accept, decline or remove any product review(s) that is not in accordance with these guidelines.


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On her official site, Meyer says she never intended to publish Bree Tanner as a stand-alone book. It was just something she'd started working on while she was still editing Eclipse. "I was thinking a lot about the newborns, imagining their side of the story, and one thing led to another." The novella came in handy during the filming of The Twilight Saga: Eclipse (in theaters June 30). Screenwriter Melissa Rosenberg, director David Slade, and actors Xavier Samuel (Riley), Bryce Dallas Howard (Victoria), and Jodelle Ferland (Bree) all read it before cameras rolled to better understand the characters.
Meyer thought the manuscript might work well as part of The Twilight Saga: The Official Guide, a tome still awaiting a pub date, but at nearly 200 pages, it was something her publisher thought worked better on its own. Meyer liked the idea of publishing it before Eclipse hits theaters, to give fans an introduction to Bree. She also considered the novella a gift to devoted Twilight readers, which is why she asked her publisher to make it available for free online.


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Good Shepherd Lutheran Church
Seeking the lost. Serving the hurting. Tending God's Flock.
1407 Cedar Ave. N.
Glencoe, MN 55336
E-mail: email@example.com
Robert Nowak, Vacancy Pastor • Ted Stroming, DCE
Sunday Festival Worship, 9 a.m.
Sunday Education, 10:15 a.m. (3 yr. old – 8th Grade, Adults)
Youth Ministry & Bible Classes (online schedule)