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Includes 1 punch for brake shoe rivets and 1 punch for clutch pulley brake. Thermostat - degree. The National Security Agency under former President Barack Obama frequently violated privacy protections while scouring through overseas intercepts — and failed to disclose the extent of the problems until late last fall, Circa reported Wednesday. These days, improvements in info technological innovation are owning broad-ranging consequences across quite a few domains of culture, and coverage makers are performing on problems involving financial efficiency, intellectual property legal rights, privateness defense, and affordability of and entry to facts. Rev Recent Clin Trials. Companies will also make you a custom alloy flywheel, you supply them a stock one, and they make an alloy center with a clutch face, and press the stock ring gear onto it from your flywheel. First, there are those vices which are connected with a particular part of the body.

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The organic phases were combined and dried for 1 h with Na 2 SO 4. The residue was then recrystallized by petroleum ether. The solvent CHCl 3 was then removed by rotary evaporation and the residue was treated by 8 mL Na 2 CO 3 50 mM , the mixture was ultra sounded for 10 min and further centrifuged at 4, rpm for 20 min.

The cationic liposomes were prepared by thin film hydration and polycarbonate membrane extrusion method according to the literatures [ 31 , 42 ]. DOX was remotely loaded into the liposomes by a transmembrane pH gradient. The resultant of multilamellar vesicles were extruded 5 times through 0. DOX concentration in the liposomes and drug encapsulation efficiency of the collected fraction were determined by measuring absorption at nm on a UV-Vis spectrophotometer PC, Shanghai Spectrum Instruments, Shanghai, China following liposome lysis in ethanol.

The samples were vortexed, and the solutions were then incubated at room temperature for 30 min to ensure siRNA loading efficiently. Free or unconjugated siRNA will migrate in the gels, but the conjugated siRNA will be stuck in the loading wells with liposomes. All the measurements were carried out at room temperature. Each parameter was measured three times, and average values and standard deviations were calculated. Agarose gel electrophoresis was used to evaluate the siRNA loading in liposomes.

Samples were observed with a fluorescence microscope Olympus, Japan. Total RNA was extracted from the cells or tissues by using Trizol reagent following the protocol suggested by the manufacturer.

The rRNA was applied as the input reference. The primers used in this study are listed as follows: Then the membranes were washed 4 times and developed by an enhanced chemiluminescence system according to the manufacturer's instructions Perkin Elmer, Waltham, MA. Cells were incubated for a further 24 h and the proportion of viable cells measured colorimetrically according to the user's manual.

Cell viability within each group was expressed as a percentage of the viability of control cells. Microscopic examination of cells or tissue section slides was performed with an Olympus SZX12 fluorescence microscope equipped with digital camera and connected to a PC running MagnaFire 2. Pictures were taken at equal exposure time for each sample. DAPI solution was employed for the staining of nucleus. Samples were observed with a fluorescence microscope Olympus, Japan and pictures were taken at same exposure time.

DOX concentration in plasma was measured by its fluorescence intensity at nm by fluorescence spectrophotometer Hitachi F, software FL solution, Japan. The concentration of DOX in each sample was calculated by using a calibration curve determined by a series of dilution of DOX solutions.

The experiment protocol was approved by the ethical committee of animal experiment of Huazhong University of Science and Technology. For the tissue distribution assay, subcutaneous KB tumors were seeded by inoculation of 2. Lung, heart, liver, spleen, kidney and tumor tissues were also collected at 3 h respectively after injection.

The concentration of DOX was determined by fluorescence spectrophotometer. DOX in plasma was measured by its fluorescence intensity at nm by fluorescence spectrophotometer Hitachi F, software FL solution, Japan. For the efficacy study, 2. The tumor sizes were measured every other day and any death of the mice was recorded. Tumor tissue uptake were assayed using the tumor tissue removed from mice for distribution study. Blood samples were collected at 24 h after the injections and then centrifuged rpm for 5 min at room temperature to remove the cells.

Then the liposomes were characterized to ensure the liposomes were prepared correctly. To find an appropriate preparation composition, we mixed the liposomes and siRNA at the ratio The result showed that when the liposomes and siRNA were combined at a ratio of This suggests that the siRNA can be fully loaded by liposomes at the ratio of This indicated that siRNA binding increased the particle size of liposomes about nm.

Results showed that all six cancer cell lines showed high level Bmi1 expression Fig. The Bmi1 protein expression in cancer or normal cell lines detected by western blot. Quantitative analysis of the western blotting bands. Hep3B cell was used as a negative control. The Hep3B cells were used as a negative control.

We first assayed the stability of siRNA conjugated with liposome in the serum. In addition, the amount of siRNA absorbed by KB cells had no significant reduction after 6 h culture compared with 1 and 3 h culture with mouse serum. As expected, free DOX showed rapid clearance from the plasma and its concentration was almost undetectable after 30 min Fig. Tissue distributions of DOX in normal and tumor tissues were observed in tumor-bearing nude mice after 3 h tail vein injection.

The pharmacokinetic analysis of DOX in mouse plasma. DOX distribution and Bmi1 expression in normal and tumor tissues in xenograft nude mice. The tissues were collected at the 3 h after the injections. Quantitative analysis DOX distribution by the microplate reader in the normal tissues of xenograft nude mice. DOX distribution by fluorescence detection in tumor tissues of mice. Total mRNA was extracted from the isolated tumor tissues 24 h after the drug injections.

At the 25th day after the injection, the mice were sacrificed and tumors were removed for volume determination and histology examination. The growth curve of tumors showed that the differences among these groups were due to continuous growth inhibition induced by these formulations Fig. To evaluate the systemic toxicity of various formulations, the body weight changes in mice were examined every 4 days after injections. At the fourth day, the DOX injected mice showed sudden weight loss compared with other group mice Fig.

The body weights of FA-DOX-L treated mice showed slight decrease, while other group mice kept slowly growing or unchanged. To determine whether the cellular apoptosis or growth inhibition was resulted from the Bmi1 expression suppression, we checked the expression of its downstream targets, p14 Arf , p16 Ink4A and p21 Cip1.

Considering the role of p14 and p21 inducing cellular apoptosis and p16 mediating cell cycle arrest, these data suggest that Bmi1 knockdown contributes as least partly to the cancer cell apoptosis and growth inhibition. Besides the efficacy of liposomal chemotherapeutic drugs, another concern about the druggability of liposomal formulation is its safety.

Excised tumors at the end point of the experiment day 24 after the drug injection. The curves present the changes of tumor sizes from the day of injection day 0. Saline injection was used as the control. The expression of Bmi1 target genes in tumor tissues when Bmi1 was knockdown. Bmi1 gene has been extensively investigated in cancer biology, with growing evidences supporting its functions in tumor formation and disease relapse in a number of cancer types.

Overexpression and activation of Bmi1 induce various forms of cancer, and are required for the onset and maintenance in various human tumors. Despite its roles in carcinogenesis, its therapeutic potential has not been adequately evaluated under a translational research setting.

A possible circumstance is that expression of the Bmi1 is also essential for the self-renew and proliferation of many types of stem cells besides the cancer cells [ 45 - 48 ], which demands a targeted delivery system to minimize the undesired side effects to these tissues. To overcome this barrier, this study was aimed at designing a targeted liposomal formulation that can specifically deliver Bmi1 siRNA and DOX into the tumor site, down regulate Bmi1 expression in the tumor and achieve therapeutic cures.

Apart from the obvious notion that cortisol is one of the major hormones responsible for glucose utilization during fasting, the question arises whether the increased stress in any way could be harmful to the human organism.

The molecular stress response in intermittently fasted subjects seems markedly different from the one associated with uncontrolled stress. In contrast, in uncontrolled stress, down regulation of the mineral corticoid receptor has been noted. Furthermore, deleterious stress responses are associated with a decrease in the expression of brain-derived neurotrophic factor BDNF , a response quite the opposite of calorie restriction and intermittent fasting, where increased concentrations of BDNF have been observed in numerous studies 4.

In conclusion, the controlled stress response from intermittent fasting seems fundamentally different from the one by uncontrolled physiological and psychological stress. Conversely, In line with the mechanisms described above, the increased stress might be one of the necessary factors for initiating molecular resistance for larger stressors, and thus promote some of the beneficial effects of intermittent fasting.

One potential serious side effect of intermittent fasting would be loss of muscle mass. Theoretically, food deprivation would result in depleted hepatic glycogen stores, leading to increased proteolysis and flux of amino acids from skeletal muscle for hepatic de novo gluconeogenesi s, to maintain healthy blood glucose concentrations.

As discussed previously though, a 24 hour short term fast is insufficient in duration to deplete liver glycogen stores in healthy subjects Up to 40 hours of total fasting does not stimulate catabolic processes and lead to skeletal muscle atrophy Modified alternate-day fasting and loss of lean body mass was investigated in only one study in the systematic search.

No loss of fat free mass in the absence of weight loss was observed compared to a control group fed a standardized diet Furthermore, an increase in ketone body concentrations has been observed in subjects on alternate-day fasting diets in both human and animal studies 17, Ketone bodies spare skeletal muscle from breakdown by providing non-glucose energy substrate for various tissues, of which the brain is the most important, and thus decrease the need for protein-derived substrates for gluconeogenetic conversion to maintain glucose homeostasis Available data thus suggests that short term fasting does not deplete hepatic glycogen stores to the extent that markedly increased proteolysis and gluconeogenesis becomes necessary to maintain healthy glucose concentrations.

Still this notion needs to be clarified in future research of longer duration. Intermittent fasting in the form of alternate day fasting in many instances reduces overall energy intake, with no obvious adverse effects, and thus becomes a model of calorie restriction in both human and animal subjects. Secondary to reduced energy intake and weight loss, effects such as reduced risk factors for cardiovascular disease, and improved glucose metabolism have been demonstrated in both animal and human subjects on true and modified alternate-day fasting diets.

In rats, protection against ischemic injury and improved survival has been demonstrated in both myocardial and cerebral ischemic events. Other beneficial effects, such as slowing the neuronal aging process and increasing cognitive functions and memory, have been observed.

Additionally, calorie restriction can reduce cancer risk and increase life span in rodent models on alternate-day fasting diets. Some effects occur even if the subject maintains body weight, suggesting that the reduced meal frequency or prolonged time in the fasted state might have some additional effects regardless of overall calorie restriction and weight loss.

In humans, modified alternate-day fasting diets might be easier to adhere to and they seemingly lead to less pronounced weight loss than true alternate-day fasting. Without causing weight loss, effects such as improved fasting insulin have been demonstrated in both animals and humans. In line with these findings, adiponectin increases in rats and humans on both true and modified alternate-day fasting diets in the absence of calorie restriction. Additionally, in mice, fat redistribution from visceral to subcutaneous stores has been observed despite unaltered overall body weight.

If this effect proves to be true in human subjects it could propose reduced disease risk despite unaltered body weight. Animal data further indicate some beneficial effects of intermittent fasting diets even without calorie restriction.

Neuronal health improvements such as resistance to excitotoxic injury have been observed. Alternate-day fasting in animals also leads to improved recovery after induced spinal cord injuries and progressive demyelinating disease of the peripheral nervous system, in the absence of calorie restriction.

Furthermore, in animal studies, changes associated with retareded tumorgenesis, such as decreased cell proliferation rates in various cell lines and decreased incidence of lymphoma, have been observed. Whether these observations are valid in human subjects as well remains an interesting area for future research. Future research is warranted to test whether the health promoting effects described in animal studies have some validity in humans.

We are in the very infancy of research on intermittent fasting in human subjects and future studies with larger sample sizes, longer durations and of better study design must be completed before any definite conclusions can be made regarding intermittent fasting and human health and the applicability to modern lifestyle.

My sincere gratitude to Staffan Lindeberg and Bengt Zöller for helping me set up the systematic search, for all the intellectually stimulating discussions and for the guidance in writing this review. The complete search term used was: Alternate day fasting and body weight, glucose metabolism and cardiovascular health in humans. Very interesting stuff, I love that even though some of the references were a bit over my head the latin names for some illnessess and the implications of these, etc most of it was completely comprehensible eventhough it was made for medical purposes.

Great summary of some of the work that has been done to date. Some more recent studies showing timing i. Keep up the nice work. Appreciate you and Berkhan sharing your work with us! Thanks for taking the time to write this up, Bojan. It was a very nice, in-depth look at the studies surrounding IF and written in a way that can appeal to the medical-savvy people, and the general public interested in health like myself. There used to be a time when people were trained to write and communicate, and from the above article this time has clearly passed.

I can see that you are using this article to try to bring people in and eventually sell them something. The way you are approaching it wont work-unless you really think people are going to read 10, words of science crap instead of just closing your site and going elsewhere. While it may seem like jargon to you, it is the preferred lexicon for scholarly publications. Do NOT presume their level of education.

I once listened to an Immunology professor at a University complain that the only articles he could understand in Science the journal were the ones on Immunology. Trust me; if you were to pick up and try to read a scholarly academic article outside of your field, you would react the way Jeff is reacting. Oh dear Jeff, you have clearly shown that this level of academia is lost on you. This is an excellent review of the available literature, written by a highly educated person. This is an extremely disrespectful comment that shows your lack of intelligence.

Think things through before you comment in future. It is Your comment that is extremely disrespectful comment and shows a lack of intelligence and knowledge of how many different styles there are in the world. I have an MS in Microbiology and have been employed as a programmer and technical writer for over 25 years. Both Jeff and Notjeff seriously? Notjeff is sadly also correct. However, Audience is Everything. If you want this information to be read and understood by people who are not your thesis committee or people with an MD or PhD in your field, you need to re-write it.

This study confirms much of the anecdotal evidence that I had already heard about the possible long-term effects of intermittent fasting. Interesting timing of this post as I was just talking with a co-worker about the benefits of intermittent fasting and the general practice of fasting. Although it has been a while many years since I have practiced fasting, it is a ritual that I have studied and used its healthy benefits in the past.

I have used short fasts of less than 36 hours and longer fasts of 7 days for personal research,purposes and practice. After having lived several years in an unhealthy way, I have re-tooled my lifestyle into a sustainable healthy way of living and have lost 55 pounds in the past year. The subject of intermittent fasting came up as I believe that I will begin a regular practice of IF as a part of this lifestyle.

I am looking forward the time when I can read your complete work as I have only scanned it this morning. Thanks for sharing your work with me. Thank you for sharing your research. The popular models of IF being 5: Do you have any results on their benefits from your research? I need to leave a quick comment just as I started reading The best and biggest recource base for the IF is by far Martin Berkhan, and his website leangains.

Now back to the reading…. It is a great summary and I do appreciate your work on this. Thanks for sharing this. It seems like most of the research in this area has been done on type two Diabetics. Ori may in fact be the first well-known proponent of IF and Warrior Diet should be recognized as one of original mainstream publications advocating IF for general health and physique improvements….

I spend much of my time researching information on fasting from peer reviewed journals through my university. I find the health benefits of IF, especially autophagy, are profound but tend to be poorly understood. I have the article you posted.

What do you know about this topic? Lots of writing but I was conflicted.. You talked in the beginning about this being your baby.. You made it sound like you had stumbled on some gold nuggets of previously unknown information. Nice resource of information overall but I still fail to understand why so many people regard IF as some unique idea or think of it as a new way to solve an old problem.

I tried it for a full year before realizing that my results were identical to when I ate 6 meals a day for a decade. Is it staving off disease in me a little longer and giving me a future benefit in that respect?

Possibly but not conclusively. It matters little and this just seems to justify that. Eat 6 meals if you like or eat 1 or 2. I can see great benefit in people linking to the article as a place to find many important studies on fasting.

Most appreciate the hard work, and you choosing to publish this free for all to read and benefit after the hours of work. Will put several links up to it. It is true that junk food can totally screw up any health benefits of IF and the people that bash it did just that, or did unhealthy low carb or too low calorie or had lots of stress in other areas of life. IF can also backfire if done to extreme.

I mean come on. Thank you for all the work of putting it together! From perspective of 1. Mood and energy 2. Long-term health and longevity 3. Physical and mental performance. All other things being equal, according to current data, should someone training two or three times a week fast once a week for the whole day or daily for about hours?

Also, assuming the three days workout regimen Mon, Wed, Fri or two days workout regimen Mon and Fri what would be the best day to fast to get most of the benefits and sacrifice as little anabolism as possible? Thank you for sharing it! Is there any scientific evidence to support the answer to this question?

What of the following types of fasting should one do if he or she is not interested in weight loss but rather muscle gain , but is very interested in all other health benefits fasting might provide and specifically in the benefit below related to glomerulosclerosis as this one can be life-saving! Intermittent fasting daily 1. Alternate day fasting 2. Rats fed on alternate days showed preserved kidney function as demonstrated by preserved glomerular filtration rate and renal plasma flow, compared to rats fed ad libitum.

Effect of long-term, alternate day feeding on renal function in aging conscious rats. Although scientific in its approach, quite accessible to the rest of us to understand and assimilate. I can usually go to somewhere between 2 -4 pm. But should I push it too long — I often do! What is causing it?! The symptoms you describe are associated with detoxing. You say the better your diet the less you experience them. The quality of your food matters, as the more toxins your body has to eliminate the more symptoms you will feel during your food-free periods.

You should be able to push through these symptoms and stretch the length of time without food. Disregard this comment, all. It seems like the carb reduction in paleo is helping. Maybe try being more strict with carb intake, 20g daily. I certainly appreciate a scientific paper on weight loss versus the advertisement-laced, fad based articles. I have read articles that state while research shows benefits for male subjects rodent and human alike , that longer periods of fasting were detrimental for females.

I came across this thesis while researching for an article I am writing about IF for women specifically. I have been practicing IF for a few years and despite many mishaps have finally hit a sweet spot with it. However, as you said have struggled to find reliable research, or in fact anything written on the subject at all.

However, this is what I have found so far:. Already lost 28 lbs and broke all my personal best in the major lifts. Thank you for sharing your hard work. Great read Bojan, thank you for posting it online.

I especially liked the section regarding the Ramadan fast. And congratulations on your MD! Oh, my blood work looks incredible as well, with my total cholesterol in the s despite very high HDL and my triglycerides in the mid 50s. Your research changes lives. This was incredibly well written, fascinating and informative. That day has finally come, it seems my fear of disappointment was just stopping me from looking a little harder.

S Ramachandran, Dawkins and the Lancet Journal. I met a trainer who has a giant of a sculpted body. He told me something about slingshot training which was based on eating and fasting cycles. He said the body releases growth hormones when we fast. Now it makes heck of a sense. Also, while fasting during Ramadan is mandatory, it is encouraged among Muslims to fast every Monday and Thursday. Very few people do it however. Like most good things in life. Fasting is prescribed to you as it was prescribed to those before you, that ye may learn self-restraint.

Thank you so much for sharing. I have been studying fasting for years on my own body as well as teaching it to others. I hope you write a book some day. I really appreciate not having to immerse myself in a stack of papers to understand some of the real science beyond IF. Just studies in rodents by now.

With increased ketones due to short period fasting and Preventing the loss of non fat mass. Can you advise a optimal period in hours of fasting followed by hours not fasting? Also on none fasting days. Would a ketogenic diet play some role in pre and post fast to minimise shock to the body and bring ketone levels up before a total nutritional cut off? And any thoughts on carb loading or cheat days relating to IF?

Why have you not had your work published in a peer reviewed journal? I find that highly suspicious. All that sounds great… but for a sec. I am from Greece and from family of building constructors. Those people were strong, lean, with stamina, poor, happy and optimistic. Now we are sick all the time, unsatisfied and with all my respect to the author work, with all that scientific data, — we forgetting that not only the quran reinforce fasting but the christianity too, vigorously i must say, — actually we exploring the proven results of fasting mechanism, without the way to reinforce the fundamental mentality of how to decide to live frugal.

Just a thought for further research: If so, why not settle on one unified term? If not, why are they used interchangeably throughout the article? Intermittent Fasting is the general classification covering all forms of short regular fasting, Alternate Day Fasting is a specific form of Intermittent Fasting where food is only eaten every second day. Thank you for sharing your work. Intermittent fasting seems to be a generally safe and effective regime; your year-long efforts help reveal the scientific underpinnings.

I was a half-assed pescatarian for many years. I went full vegan in March Going vegan was hard, especially the beginning. In March I was around My partner gets home late, loves to cook and thinks nothing of eating a gigantic steak and full meal at 11pm. No first aid kit is available. Your priority is to get the person to a hospital at a distance of 2 hours. The wound is bleeding and needs to be kept clean and bacteria free till the person reaches the hospital.

What will you do? Think of a solution. It is quite close to you. With all due respect, Professor Boyd, your argument is not at all compelling.

It seems that you are taking the "thinking outside the box" TOTB metaphor much more literally than it is intended or, at least, as I and may others infer. To refer to TOTB as "dangerous" is naive, at best. I, personally, have seen the positive, tranformative effects of not only the 9-dots exercise, but also the occasional use of the term to remind individuals after-the-fact about the value of thinking differently.

The experiment you refer to doesn't even come close to proving what you suggest that it does. To use the term "proving" in an argument like this is laughable. In real life, you absolutely WILL find boxes Here, the term is not literal; rather, it refers to a mindset, a perspective, a belief, or an assumption. It is precisely how the human mind works.

We all think in boxes all the time. The "sin," if you will, is not in thinking inside of a box A different -- and very healthy, positive, and productive -- way to think about TOTB is to understand that it merely represents an insight that can remind an individual to consciously become aware of limiting assumptions.

And, upon such awareness, to open ones mind and imagination to actively explore new possibilities beyond the obvious or initial answer. If you don't regard this as valid contribution to creativity, then I suggest you consider spending a bit more time outside of that "box" that you've presented here. I couldn't have said it any better TOTB is a beautiful skill to have.

We are born into multiple boxes that are created upon social agreements e. Saying that TOTB is a negative thing is a very conservative statement and someone who has such a belief is scared of change, scared of diversity and scared of anything that is abstract and out of order.

I think much of which is vaguely labeled "creative thinking" is actually the result of healthy skepticism. We should make a habit of challenging conventional wisdom and the way things have always been done. On encountering the conventional approach, our attitude should be doubtful: Fold the paper so all the dots ovelap. Use four lines to connect four dots. Hold the folded paper up to the light For that matter, you could fold the paper until all the dots overlapped and you would not need to waste any pencil lead; Thinking outside The Box.

Use a very wide pencil lead or charcoal block for that matter, connect all the dots in one fell swoop; Thinking outside The Box. Forego a pencil altogether and use a bucket of paint to create a huge blot over all the dots; Thinking outside The Box. Question the dots and why they need to be connected in the first place; Thinking outside The Box. Erase the dots; they are a distraction to Thinking outside The Box. Create your own dots and lines in any fashion you desire; Thinking outside The Box.

I play chess with my pc, an beat it all the time, and the reasoning is I do not think logically, like the pc does. Also when you go the a school that teaches how to think about something, that is all you know how to do. I have had engineers come to my deck, hand me a set of blueprints, because that was the way they were taught. They are never taught to look at it, in there mind to see it working. What I do is show them how wrong they are, an ask them what tool in the world can cut a square hole inside the middle of two long tubes.

If was going to tell you about an airplane the TR-3B, it travels a little bit under light speed, an it uses nuclear fusion, which turns into plasma an powers the craft, that was built outside the box. You my brother, do not have the inkling of understanding to think outside the box. Japanese traders have found that candlesticks form interesting patterns in the seeming market chaos.

The "Forex Candlestick Tactics" ebook reveals powerful methods to trade candlestick patterns combined with standard technical indicators. Check out what's inside:. Structured tools boost, not limit, our creative output. New research suggests we have a tendency to look like our names. Back Find a Therapist. Young People Taking on a Larger Role.

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