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It's estimated that nearly half of American grownups attempt to slim down each year.
Among the very best methods to lose weight is by changing your diet.
Yet, the large variety of offered diet plans may make it tough to begin, as you're uncertain which one is most suitable, sustainable, and reliable.
Some diet plans aim to suppress your cravings to reduce your food intake, while others suggest restricting your consumption of calories and either carbohydrates or fat.
What's more, many offer health benefits that go beyond weight-loss.
Here are the 8 finest diet plans to assist you shed weight and improve your overall health.
1. Periodic fasting Fat Loss Homeopathy
Intermittent fasting is a dietary strategy that cycles in between durations of fasting and eating.
Different types exist, including the 16/8 method, which involves restricting your calorie consumption to 8 hours each day, and the 5:2 approach, which restricts your everyday calorie intake to 500– 600 calories twice weekly.
How it works: Periodic fasting restricts the time you're permitted to consume, which is a basic way to lower your calorie intake. This can result in weight reduction– unless you compensate by eating excessive food throughout permitted consuming periods.
Weight loss: In an evaluation of research studies, periodic fasting was shown to trigger 3– 8% weight loss over 3– 24 weeks, which is a substantially greater percentage than other approaches .
The same review revealed that by doing this of consuming might decrease waist area by 4– 7%, which is a marker for damaging stomach fat.
Other studies discovered that periodic fasting can increase fat loss while maintaining muscle mass, which can enhance metabolic process.
Other benefits: Periodic fasting has actually been connected to anti-aging results, increased insulin sensitivity, improved brain health, minimized swelling, and many other benefits.
Disadvantages: In general, intermittent fasting is safe for a lot of healthy grownups.
That stated, those conscious drops in their blood glucose levels, such as some individuals with diabetes, low weight, or an eating condition, in addition to pregnant or breastfeeding women, should talk to a health professional before starting intermittent fasting.
Intermittent fasting cycles in between durations of fasting and consuming. It has been revealed to aid weight loss and is linked to lots of other health benefits.
2. Plant-based diets
Plant-based diet plans may assist you drop weight. Vegetarianism and veganism are the most popular versions, which limit animal products for health, ethical, and environmental factors.
Nevertheless, more versatile plant-based diet plans likewise exist, such as the flexitarian diet, which is a plant-based diet that allows eating animal products in moderation.
How it works: There are many kinds of vegetarianism, but many include eliminating all meat, poultry, and fish. Some vegetarians might similarly prevent eggs and dairy.
The vegan diet takes it an action further by restricting all animal products, along with animal-derived items like dairy, gelatin, honey, whey, casein, and albumin.
There are no well-defined guidelines for the flexitarian diet plan, as it's a lifestyle modification instead of a diet plan. It encourages consuming mostly fruits, veggies, vegetables, and entire grains however allows for protein and animal items in moderation, making it a popular option.
A number of the limited food groups are high in calories, so restricting them might assist weight-loss.
Weight reduction: Research shows that plant-based diets are effective for weight reduction .
A review of 12 research studies consisting of 1,151 participants discovered that individuals on a plant-based diet lost an average of 4.4 pounds (2 kg) more than those who consisted of animal products .
Plus, those following a vegan diet lost an average of 5.5 pounds (2.5 kg) more than individuals not eating a plant-based diet plan .
Plant-based diets likely aid weight reduction because they tend to be rich in fiber, which can help you stay fuller for longer, and low in high-calorie fat.
Other benefits: Plant-based diets have been connected to many other benefits, such as a minimized risk of chronic conditions like heart problem, certain cancers, and diabetes. They can likewise be more environmentally sustainable than meat-based diets.
Downsides: Though plant-based diet plans are healthy, they can restrict important nutrients that are usually discovered in animal products, such as iron, vitamin B12, vitamin D, calcium, zinc, and omega-3 fats.
A flexitarian method or correct supplementation can help account for these nutrients.
Plant-based diet plans restrict meat and animal items for various factors. Research studies show that they aid weight loss by lowering your calorie consumption and use numerous other advantages.
3. Low-carb diets
Low-carb diet plans are among the most popular diet plans for weight-loss. Examples consist of the Atkins diet plan, ketogenic (keto) diet plan, and low-carb, high-fat (LCHF) diet.
Some varieties lower carbohydrates more dramatically than others. For example, very-low-carb diets like the keto diet plan limit this macronutrient to under 10% of overall calories, compared to 30% or less for other types .
How it works: Low-carb diet plans limit your carb intake in favor of protein and fat.
They're generally higher in protein than low-fat diets, which is necessary, as protein can help curb your appetite, raise your metabolic process, and conserve muscle mass.
In very-low-carb diet plans like keto, your body begins utilizing fats rather than carbohydrates for energy by converting them into ketones. This process is called ketosis.
Weight-loss: Numerous research studies indicate that low-carb diet plans can assist weight reduction and may be more efficient than traditional low-fat diet plans.
For instance, a review of 53 research studies consisting of 68,128 individuals found that low-carb diet plans resulted in considerably more weight reduction than low-fat diets.
What's more, low-carb diet plans appear to be quite efficient at burning hazardous tummy fat .
Other benefits: Research study suggests that low-carb diets might lower risk aspects for heart disease, consisting of high cholesterol and blood pressure levels. They may also improve blood glucose and insulin levels in individuals with type 2 diabetes.
Downsides: In many cases, a low-carb diet plan might raise LDL (bad) cholesterol levels. Very-low-carb diet plans can likewise be hard to follow and cause indigestion in some people.In very rare circumstances, following a very-low-carb diet plan may cause a condition called ketoacidosis, an unsafe metabolic condition that can be fatal if left neglected (32Trusted Source, 33Trusted Source).
Low-carb diets restrict your carb intake, which motivates your body to utilize more fat as fuel. They can help you reduce weight and use numerous other benefits.
4. The paleo diet
The paleo diet plan advocates consuming the exact same foods that your hunter-gatherer forefathers presumably ate.
It's based on the theory that modern-day illness are linked to the Western diet, as advocates believe that the body hasn't evolved to process vegetables, grains, and dairy.
How it works: The paleo diet plan advocates consuming whole foods, fruits, vegetables, lean meats, nuts, and seeds. It limits the intake of processed foods, grains, sugar, and dairy, though some less restrictive versions enable some dairy products like cheese.
Weight loss: Many studies have revealed that the paleo diet can assist weight loss and reduce harmful belly fat.
For instance, in one 3-week study, 14 healthy grownups following a paleo diet plan lost an average of 5.1 pounds (2.3 kg) and lowered their waist circumference– a marker for belly fat– by approximately 0.6 inches (1.5 cm) .
Research study also recommends that the paleo diet plan might be more filling than popular diet plans like the Mediterranean diet plan and low-fat diet plans. This may be due to its high protein material.
Other benefits: Following the paleo diet may decrease a number of heart disease threat aspects, such as hypertension, cholesterol, and triglyceride levels (40Trusted Source, 41Trusted Source).
Disadvantages: Though the paleo diet is healthy, it limits numerous healthy food groups, including beans, whole grains, and dairy.
The paleo diet supporters eating whole foods, likewise to how your forefathers consumed. Research studies show that it might assist weight loss and decrease heart problem danger factors.
5. Low-fat diets
Like low-carb diet plans, low-fat diet plans have actually been popular for decades.
In general, a low-fat diet involves restricting your fat intake to 30% of your everyday calories.
Some very- and ultra-low-fat diet plans intend to restrict fat consumption to under 10% of calories.
How it works: Low-fat diets restrict fat intake since fat offers about twice the variety of calories per gram, compared with the other two macronutrients– protein and carbohydrates.
Ultra-low-fat diet plans include fewer than 10% of calories from fat, with around 80% of calories coming from carbohydrates and 10% from protein.
Ultra-low-fat diets are mainly plant-based and limit meat and animal products.
Weight loss: As low-fat diets limit calorie consumption, they can help weight loss.
An analysis of 33 research studies consisting of over 73,500 participants found that following a low-fat diet plan led to little but relevant modifications in weight and waist circumference.
Nevertheless, while low-fat diets seem as efficient as low-carb diet plans for weight loss in controlled circumstances, low-carb diet plans seem to be more efficient day to day.
Ultra-low-fat diet plans have actually been revealed to be effective, especially among individuals with weight problems. For example, an 8-week study in 56 participants discovered that consuming a diet plan consisting of 7– 14% fat caused an average weight reduction of 14.8 pounds (6.7 kg).
Other advantages: Low-fat diets have been linked to a decreased risk of heart problem and stroke. They may likewise decrease swelling and improve markers of diabetes.
Disadvantages: Restricting fat excessive can result in health issue in the long term, as fat plays a key role in hormonal agent production, nutrient absorption, and cell health. Furthermore, very-low-fat diet plans have actually been connected to a higher danger of metabolic syndrome.
Low-fat diets limit your consumption of fat, as this macronutrient is higher in calories than protein and carbohydrates. Research studies have linked low-fat diet plans to weight-loss and lower dangers of cardiovascular disease and diabetes.
6. The Mediterranean diet plan
The Mediterranean diet plan is based on foods that people in countries like Italy and Greece utilized to consume.
Though it was created to lower heart disease risk, various research studies show that it can also help weight reduction.
How it works: The Mediterranean diet advocates eating lots of fruits, veggies, nuts, seeds, legumes, bulbs, entire grains, fish, seafood, and extra virgin olive oil.
Foods such as poultry, eggs, and dairy items are to be consumed in small amounts. On the other hand, red meats are restricted.
Additionally, the Mediterranean diet plan restricts fine-tuned grains, trans fats, improved oils, processed meats, sugarcoated, and other extremely processed foods.
Weight reduction: Though it's not specifically a weight-loss diet, lots of studies show that adopting a Mediterranean-style diet plan may help weight-loss.
For example, an analysis of 19 research studies found that individuals who integrated the Mediterranean diet plan with exercise or calorie restriction lost an average of 8.8 pounds (4 kg) more than those on a control diet plan.
Other benefits: The Mediterranean diet encourages consuming a lot of antioxidant-rich foods, which might assist fight inflammation and oxidative tension by neutralizing free radicals. It has actually been connected to reduced threats of heart disease and premature death.
Disadvantages: As the Mediterranean diet plan is not strictly a weight loss diet, individuals might not reduce weight following it unless they likewise take in fewer calories.
The Mediterranean diet stresses consuming a lot of fruits, veggies, fish, and healthy oils while restricting refined and highly processed foods. While it's not a weight loss diet, research studies show that it can promote weight-loss and general health.
7. WW (Weight Watchers).
WW, previously Weight Watchers, is among the most popular weight loss programs worldwide.
While it doesn't limit any food groups, individuals on a WW plan must consume within their set everyday points to reach their perfect weight.
How it works: WW is a points-based system that designates various foods and drinks a value, depending on their calorie, fat, and fiber contents.
To reach your desired weight, you need to stay within your daily point allowance.
Weight loss: Many studies reveal that the WW program can assist you slim down.
For example, an evaluation of 45 studies discovered that individuals who followed a WW diet lost 2.6% more weight than individuals who got basic therapy.
What's more, people who follow WW programs have been shown to be more successful at maintaining weight-loss after several years, compared with those who follow other diets.
Other advantages: WW allows flexibility, making it easy to follow. This makes it possible for individuals with dietary limitations, such as those with food allergies, to follow the plan.
Disadvantages: While it allows for flexibility, WW can be expensive depending on the subscription plan. Likewise, it's flexibility can be a failure if dieters choose unhealthy foods.
WW, or Weight Watchers, is a weight-loss program that utilizes a points-based system. Studies show that it's effective for long-lasting weight-loss and highly versatile.
8. The DASH diet plan.
Dietary Approaches to Stop High Blood Pressure, or DASH diet, is an eating strategy that is created to assist treat or avoid hypertension, which is scientifically known as high blood pressure.
It highlights eating plenty of fruits, vegetables, whole grains, and lean meats and is low in salt, red meat, sugarcoated, and fat.
While the DASH diet plan is not a weight-loss diet, many individuals report dropping weight on it.
How it works: The DASH diet suggests specific servings of different food groups. The number of portions you are enabled to consume depends upon your everyday calorie consumption.
For instance, a typical person on the DASH diet would eat about 5 servings of veggies, 5 servings of fruit, 7 portions of healthy carbs like whole grains, 2 servings of low-fat dairy products, and 2 servings or fewer of lean meats each day.
In addition, you're permitted to eat nuts and seeds 2– 3 times per week.
Weight-loss: Research studies reveal that the DASH diet can help you drop weight .
For example, an analysis of 13 studies discovered that individuals on the DASH diet lost significantly more weight over 8– 24 weeks than people on a control diet plan.
Other benefits: The DASH diet plan has been revealed to minimize blood pressure levels and several heart disease threat aspects. Likewise, it might help fight persistent depressive signs and lower your threat of breast and colorectal cancer.
Downsides: While the DASH diet might help weight-loss, there is mixed evidence on salt intake and blood pressure. In addition, eating insufficient salt has been linked to increased insulin resistance and an increased danger of death in people with heart failure.
The DASH diet is a low-salt diet plan that has actually been revealed to help weight-loss. Research studies have also linked it to fringe benefits for your heart and lowered risks of other persistent illness.
The bottom line. Fat Loss Homeopathy
Many diets can help you drop weight.
Some of the most well-researched diet plans and consuming plans consist of periodic fasting, plant-based diet plans, low-carb diets, low-fat diet plans, the paleo diet, the Mediterranean diet plan, WW (Weight Watchers), and the DASH diet plan.
While all of the above diets have actually been shown to be reliable for weight reduction, the diet you choose ought to depend upon your way of life and food choices. This ensures that you are more likely to stick to it in the long term.


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subset: commoncrawl


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Red wine koszyki has actually been actually looked at as being one of the best types of a glass of wine, which is likewise understood to be filled with anti-oxidants as well as vitamins. Wine is basically a cabernet produced by utilizing reddish grape varieties in their cultivation procedure. The real color of this particular red or white wine may go up from a dull violet, popular of young wines, throughout to black, which is actually the most popular type of red wine.
The greatest method to consume the greatest reddish a glass of wine is actually by drinking all of them in the glass directly out of the container, without ice or any type of ice dices. Some people like not to combine the ice along with the taste of red a glass of wine, while some alcoholic beverage along with the ice to maintain the cold sensation of the red white wine and also enjoy the taste.
If you are actually questioning just how to consume your beloved form of red wine, after that you can easily learn to perform it on your own. Having said that, if you don't possess any type of knowledge in accomplishing this, after that you ought to head to your local pub and order the exact same. The bartender will show you all the necessary tromp just how to enjoy your preferred form of cabernet as well as have the capacity to do it quickly at home.
When delighting in red white wine at house, it is actually essential to keep your red wine glasses neat and also tidy. You ought to keep your glass upright and with a stem to stop some spillages when you are actually consuming alcohol.
After you have actually finished consuming your white wine, see to it that you rinse the a glass of wine out of your mouth prior to you brush your teeth. This is actually to prevent the stain coming from getting along your teeth. Use the back of your hand to rinse your mouth carefully along with water as well as brush the stain away from your mouth. This is actually likewise a great tip if you take place to spill your beverage.
Just before wine gift you wash down your red wine glass along with water, put a little of it and also put it inside a glass of water, to make sure that you receive the taste and also give off the cabernet. just before you eat it. It's encouraged to carry out it a couple times and also view whether the flavor as well as odor are actually still certainly there or not.
It is actually essential that you perform certainly not consume very much at once when you are drinking red a glass of wine. It is actually feasible that you may certainly not manage to complete the whole container or the entire glass. If you want to appreciate a good taste of red wine, then it is actually well to maintain it in your mouth for a longer opportunity, certainly not just half an hour or so.
By stashing it in a glass of water and also leaving it there for a day, at that point you obtain the flavor of reddish white wine and certainly not merely consuming it directly from the bottle. Once you have actually sampled the new palate of the reddish wine, you will certainly certainly never yearn for to obtain cold bottles of red wine once more.
Red wine Pinot Noir is actually a range of red grape varietals produced by making fermented grape extract coming from reddish grape skin as well as skin layers. The different colors of this a glass of wine can differ coming from strong purple, often connected with young merlots, approximately medium reddish for more mature reddishes, as well as red via to brownish for aged reddishes. The key explanation for the assorted shades of the assortment of wine derives from the way the grapes are actually developed, rather than its flavor.
Red wine can easily be actually found in all components of the world, although it is very most well-liked in Northern Europe as well as Northern America. Reddish a glass of wine is certainly not as widely made use of in Asia, it is actually made use of there in numerous different means.
One of the greatest tastes associated with red white wine is the tannin. Tannin in red white wine is actually normally located in red glass of wines grown in warmer temperatures such as in North United States, South The United States, as well as Eastern Europe.
Merlot contains tannic acid that is not just located in huge quantities in reddish grapes, however additionally in some red berries, dark berries, raisins, cherries, black currants, as well as raspberries. While the level of acidity of these fruit products contributes to the flavor of wine, the acid itself is necessary to some folks, as it sends out an uncomfortable odor when it is actually corroded in the oral cavity.
Wine, although it has a wonderful wide array of preferences, originates from a restricted variety of grapes. The grapes made use of in creating merlot contain every one of the different kinds of glucoses essential to exchange booze, featuring the tannin and the acids.


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50 Small Living Room Design Ideas to Copy Right Now! – Sharp Aspirant
Small living room design ideas. If you're looking for some inspo to redesign your usual boring living room, then you're in the right place! Whether your style is bohemian, eclectic, modern, scandinavian, traditional, farmhouse, or contemporary we have something for you here!
Shared by: karenores


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  How To Lose Weight Fast - Just Got Easier - Weight Loss
- An Interesting Topic, Negative Calories.
The negative calories are real and will help you achieve max calories to lose weight c2h and how to lose weight fast . Nothing could be simpler than this.
As you eat, you must be using calories. Eating and digesting is itself an activity, and those activities require calories. Negative calorie foods take a lot to digest but only offer a few calories to your body. This is important for max calories to lose weight c2h and how to lose weight fast .
Calories that are in the fat can be used for this. A pretty cool fact, eh? Study this idea for a minute. Just by eating, again, just by eating, you accomplished weight loss.
Snacks have been many folks downfall in the weight gain arena. Imagine, though if all the snacks you ate were negative calorie foods? Day or night, enjoy negative calorie foods.
The quite common negative calorie foods include celery and carrots.
- Quantity Of Meals In One Day.
It is not like you are climbing a mountain to achieve max calories to lose weight c2h and how to lose weight fast . Knowledge of this will be a sure way to get results, because it really is not a difficult task to lose weight.
Skip meals and you will not optimize your weight loss. Truly. A totally unexpected statement? Having breakfast is of utmost importance.
Breakfast is the start of the day and it is the ending of a, fast. So the phase of doing without food is over. We want to get rid of calories from fat. Metabolism will do that, and we eat to get that going.
Have a greater number of meals than usual (maybe 5 to 6), but make them smaller meals. Okay, so what is this called? Grazing. We want to consume calories from fat, so we eat to get the metabolism to do just that. We go forward this way and thereby accomplish max calories to lose weight c2h and how to lose weight fast .
- Food Calories Examples.
We will keep in the topic area of max calories to lose weight c2h and how to lose weight fast . If you want to lose weight you have to know this info. Food labels tell an important story. A picture is worth a thousand words. Lets see an example.
A comparison can be quite dramatic. Try this one out for size. 1 cup cooked green beans contains just a wee bit of calories, 38 calories, but when you go ahead and compare that to 1 cup cooked lima beans it has a whopping 260 calories.
Ponder this for while before you move on. Which one do you want in your diet, starting today? If you just added the limas to your diet today it could mean adding 2 or 3 pounds to your weight a month.
For another example, compare a sliced raw apple with a slice of apple pie. The apple has 60 calories. The apple pie has 400 calories.
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text: Rebeccagrace (rebeccagrace) wrote in vaginapagina, 2014-04-12 00:50:00 Rebeccagrace rebeccagrace vaginapagina 2014-04-12 00:50:00 Previous Share Flag Next Forgot what day I was on... Hello, VP. I am recently divorced and so have decided to date freely, which means I have several fwbs. Last month with a guy the condom broke and I had to take plan B. Well this time a guy came over and before we got started I asked him if he brought a condom, but he didn't because he came straight from work, so me, being moderately proficient in the fam method, decided, oh, I just got off my period, I'm fine. So I let him finish inside me. Afterwards I was randomly thinking and realized I'm on day 12 of a 30 day cycle. I already took plan B last month and I'm not wanting to take it again. I also am not on any birth control. How likely is it that I could get pregnant from sex at this time? I can't believe I miscalculated my cycle day so epically! It wouldn't be the end of the world but a pregnancy would...complicate things immensely. We barely know each other at all and just...no. So in your expert opinions...how likely could it be that this encounter will result in a pregnancy? Subscribe Post a new comment Error Anonymous comments are disabled in this journal We will log you in after post We will log you in after post We will log you in after post We will log you in after post We will log you in after post Anonymously switch LiveJournal Facebook Twitter OpenId Google MailRu VKontakte Anonymously default userpic Your reply will be screened Your IP address will be recorded Post a new comment Preview comment Help 12 comments Post a new comment 12 comments


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Cholesterol is an essential component of mammalian cell membranes, constituting up to 50% of plasma membrane lipids. By contrast, it accounts for only 5% of lipids in the endoplasmic reticulum (ER)1. The ER enzyme sterol O-acyltransferase 1 (also named acyl-coenzyme A:cholesterol acyltransferase, ACAT1) transfers a long-chain fatty acid to cholesterol to form cholesteryl esters that coalesce into cytosolic lipid droplets. Under conditions of cholesterol overload, ACAT1 maintains the low cholesterol concentration of the ER and thereby has an essential role in cholesterol homeostasis2,3. ACAT1 has also been implicated in Alzheimer's disease4, atherosclerosis5 and cancers6. Here we report a cryo-electron microscopy structure of human ACAT1 in complex with nevanimibe7, an inhibitor that is in clinical trials for the treatment of congenital adrenal hyperplasia. The ACAT1 holoenzyme is a tetramer that consists of two homodimers. Each monomer contains nine transmembrane helices (TMs), six of which (TM4-TM9) form a cavity that accommodates nevanimibe and an endogenous acyl-coenzyme A. This cavity also contains a histidine that has previously been identified as essential for catalytic activity8. Our structural data and biochemical analyses provide a physical model to explain the process of cholesterol esterification, as well as details of the interaction between nevanimibe and ACAT1, which may help to accelerate the development of ACAT1 inhibitors to treat related diseases.
AssuntosMicroscopia Crioeletrônica , Esterol O-Aciltransferase/química , Esterol O-Aciltransferase/ultraestrutura , Ureia/análogos & derivados , Colesterol/química , Colesterol/metabolismo , Histidina/química , Histidina/metabolismo , Holoenzimas/química , Holoenzimas/ultraestrutura , Humanos , Ligantes , Modelos Moleculares , Multimerização Proteica , Eletricidade Estática , Ureia/química
Triacylglycerols store metabolic energy in organisms and have industrial uses as foods and fuels. Excessive accumulation of triacylglycerols in humans causes obesity and is associated with metabolic diseases1. Triacylglycerol synthesis is catalysed by acyl-CoA diacylglycerol acyltransferase (DGAT) enzymes2-4, the structures and catalytic mechanisms of which remain unknown. Here we determined the structure of dimeric human DGAT1, a member of the membrane-bound O-acyltransferase (MBOAT) family, by cryo-electron microscopy at approximately 3.0 Å resolution. DGAT1 forms a homodimer through N-terminal segments and a hydrophobic interface, with putative active sites within the membrane region. A structure obtained with oleoyl-CoA substrate resolved at approximately 3.2 Å shows that the CoA moiety binds DGAT1 on the cytosolic side and the acyl group lies deep within a hydrophobic channel, positioning the acyl-CoA thioester bond near an invariant catalytic histidine residue. The reaction centre is located inside a large cavity, which opens laterally to the membrane bilayer, providing lipid access to the active site. A lipid-like density-possibly representing an acyl-acceptor molecule-is located within the reaction centre, orthogonal to acyl-CoA. Insights provided by the DGAT1 structures, together with mutagenesis and functional studies, provide the basis for a model of the catalysis of triacylglycerol synthesis by DGAT.
AssuntosBiocatálise , Microscopia Crioeletrônica , Diacilglicerol O-Aciltransferase/metabolismo , Diacilglicerol O-Aciltransferase/ultraestrutura , Triglicerídeos/biossíntese , Acil Coenzima A/química , Acil Coenzima A/metabolismo , Acil Coenzima A/ultraestrutura , Aciltransferases/química , Aciltransferases/metabolismo , Domínio Catalítico , Membrana Celular/química , Membrana Celular/metabolismo , Diacilglicerol O-Aciltransferase/química , Histidina/química , Histidina/metabolismo , Humanos , Interações Hidrofóbicas e Hidrofílicas , Modelos Moleculares , Multimerização Proteica , Especificidade por Substrato
Lung adenocarcinoma (LUAD) is the most common subtype of nonsmall cell lung cancer, and 5-year survival rate is only 15% in recent years. This study aimed to explore the FAM83A expression and its potential functions in LUAD. Data of LUAD were downloaded from The Cancer Genome Atlas database. Expression level of FAM83A was compared between LUAD samples and adjacent normal samples. The association between FAM83A expression and clinic-pathological parameters was analyzed, as well as copy number variation and methylation status. Kaplan-Meier curve was used to visualize the relationship of FAM83A expression with survival outcomes. Finally, gene set enrichment analysis was used to identify potential signaling pathways in LUAD specimens. FAM83A expression was significantly correlated with four clinical factors in LUAD specimens, age, gender, smoking, and overall survival status (all p < 0.05). High expression level of FAM83A was negatively correlated with methylation level. Moreover, patients in low expression groups exhibited a better prognosis than those in high expressed groups, which was independent of gender (p < 0.001). Histidine metabolism pathway was significantly upregulated in FAM83A-high expressed samples than FAM83A-low expressed samples according to functional enrichment analysis. High expression of FAM83A predicted a poor prognosis in LUAD patients. Our study demonstrated that FAM83A might be a potential biomarker and meaningful therapeutic target in LUAD.
AssuntosAdenocarcinoma de Pulmão/diagnóstico , Adenocarcinoma de Pulmão/genética , Proteínas de Neoplasias/genética , Oncogenes/genética , Adenocarcinoma de Pulmão/metabolismo , Adenocarcinoma de Pulmão/patologia , Idoso , Carcinogênese/genética , Proliferação de Células/genética , Metilação de DNA/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Histidina/metabolismo , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Prognóstico
NifB is a radical S-adenosyl-L-methionine (SAM) enzyme that is essential for nitrogenase cofactor assembly. Previously, a nitrogen ligand was shown to be involved in coupling a pair of [Fe4S4] clusters (designated K1 and K2) concomitant with carbide insertion into an [Fe8S9C] cofactor core (designated L) on NifB. However, the identity and function of this ligand remain elusive. Here, we use combined mutagenesis and pulse electron paramagnetic resonance analyses to establish histidine-43 of Methanosarcina acetivorans NifB (MaNifB) as the nitrogen ligand for K1. Biochemical and continuous wave electron paramagnetic resonance data demonstrate the inability of MaNifB to serve as a source for cofactor maturation upon substitution of histidine-43 with alanine; whereas x-ray absorption spectroscopy/extended x-ray fine structure experiments further suggest formation of an intermediate that lacks the cofactor core arrangement in this MaNifB variant. These results point to dual functions of histidine-43 in structurally assisting the proper coupling between K1 and K2 and concurrently facilitating carbide formation via deprotonation of the initial carbon radical.
AssuntosProteínas de Bactérias/metabolismo , Methanosarcina/metabolismo , Nitrogênio/metabolismo , Nitrogenase/biossíntese , Alanina/genética , Alanina/metabolismo , Proteínas de Bactérias/genética , Espectroscopia de Ressonância de Spin Eletrônica , Histidina/genética , Histidina/metabolismo , Ligantes , Methanosarcina/genética , Mutagênese , Nitrogenase/genética , Espectroscopia por Absorção de Raios X
Cloning and expression of a desired gene is indispensable in molecular biology studies. Expression vectors, in this regard, should offer much needed flexibility and choice of cloning strategies for both in vivo and in vitro protein expression experiments. Furthermore, availability of option to choose from various reporter tags allows one to be flexible during designing of an experiment in a more relevant manner. Thus, the need of a versatile expression system cannot be ignored. Although several different expression vectors are available for gene expression in mycobacteria, they lack the required versatility of expression and the inclusion of reporter tags. We here present the construction of a set of nine E. coli-Mycobacterium shuttle plasmids, which offer a combination of three mycobacterial promoter systems (heat shock inducible-hsp60, tetracycline-, and acetamide-inducible) along with three polypeptide tags (Green Fluorescent Protein (GFP), Glutathione S-transferase (GST) and hexa-histidine tag). These vectors offer the cloning of a target gene in all the nine given vectors in parallel, thus allowing the generation of recombinant plasmids that will express the target gene from different promoters with different tags. Here, while the hexa-histidine and GST tags can be used for protein purification and pull-down experiments, the GFP-tag can be used for protein localization within the cell. Additionally, the vectors also offer the choice of positioning of the reporter tag either at the N-terminus or at the C-terminus of the expressed protein, which is achieved by cloning of the gene at any of the two blunt-end restriction enzyme sites available in the vector. We believe that these plasmids will be extremely useful in the gene expression studies in mycobacteria by offering the choices of promoters and reporters. Our work also paves the way to developing more such plasmids with other tags and promoters that may find use in mycobacterial biology.
AssuntosEngenharia Genética/métodos , Vetores Genéticos/genética , Mycobacterium/genética , Escherichia coli/genética , Genes Reporter , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Histidina/genética , Histidina/metabolismo , Oligopeptídeos/genética , Oligopeptídeos/metabolismo , Regiões Promotoras Genéticas , Ativação Transcricional
To formulate low-protein diets for blue foxes with sufficient amounts of amino acids (AA), AA digestibility and AA requirements of the animals are crucial information. Therefore, a digestibility and nitrogen (N) balance trial was conducted with 20 blue foxes to determine the macronutrient and AA digestibility and N utilisation in low-protein diets supplemented with DL-methionine (Met) and L-histidine (His). In addition, plasma urea and plasma AA were measured. The diets were designated as P24 (control), P20, P20M, P16M and P16MH and contained energy from digestible crude protein (DCP) at 24%, 20% or 16% of total dietary metabolisable energy (ME). The 20% protein level was fed with or without Met and the 16% protein level was fed with Met and with or without His. The apparent total-tract digestibility (ATTD) of crude protein linearly decreased with decreasing dietary protein level. The ATTD of dry matter, organic matter and crude carbohydrates increased when wheat starch was added as a replacement for protein. The apparent ileal digestibility (AID) and ATTD methods were compared to determine the AA digestibility. The decreasing dietary protein supply decreased the ATTD of most of the AA: threonine, tryptophan (Trp), valine, alanine (Ala), aspartic acid (Asp), glutamic acid, glycine (Gly), proline (Pro), serine (Ser) and total AA. The AID of the AA was constant between diets. Diverging AA showed higher or lower digestibility when determined in the AID or ATTD methods. Isoleucine, lysine, Met, Ala and tyrosine showed higher levels of AID. Arginine, His, cysteine (Cys), Trp, Asp, Gly, Pro and Ser showed higher levels of ATTD, which may reflect the net loss of these AA in the large intestine. Met and His supplementation improved the ATTD and AID of the AA in question, respectively, but did not affect the other variables examined. N retention did not differ between diets and renal N excretion decreased with decreasing protein level; thus N utilisation improved. It was concluded that the protein supply and AA composition in low-protein diets with supplemented Met were adequate for adult blue foxes, since the lower protein supply improved N utilisation and did not affect N retention. However, His supplementation failed to reach the designed level and therefore showed no clear results.
AssuntosDieta com Restrição de Proteínas/veterinária , Digestão , Raposas/fisiologia , Histidina/metabolismo , Íleo/fisiologia , Nitrogênio/metabolismo , Racemetionina/metabolismo , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal/efeitos dos fármacos , Animais , Suplementos Nutricionais/análise , Digestão/efeitos dos fármacos , Histidina/administração & dosagem , Masculino , Racemetionina/administração & dosagem , Distribuição Aleatória
Manipulation of circulating histidine-containing dipeptides (HCD) has been shown to affect the development of diabetes and early-stage diabetic nephropathy (DN). The aim of the present study was to investigate whether such interventions, which potentially alter levels of circulating HCD, also affect the development of advanced-stage DN. Two interventions, aerobic exercise training and overexpression of the human carnosinase-1 (hCN1) enzyme, were tested. BTBR ob/ob mice were either subjected to aerobic exercise training (20 wk) or genetically manipulated to overexpress hCN1, and different diabetes- and DN-related markers were compared with control ob/ob and healthy (wild-type) mice. An acute exercise study was performed to elucidate the effect of obesity, acute running, and hCN1 overexpression on plasma HCD levels. Chronic aerobic exercise training did not affect the development of diabetes or DN, but hCN1 overexpression accelerated hyperlipidemia and aggravated the development of albuminuria, mesangial matrix expansion, and glomerular hypertrophy of ob/ob mice. In line, plasma, kidney, and muscle HCD were markedly lower in ob/ob versus wild-type mice, and plasma and kidney HCD in particular were lower in ob/ob hCN1 versus ob/ob mice but were unaffected by aerobic exercise. In conclusion, advanced glomerular damage is accelerated in mice overexpressing the hCN1 enzyme but not protected by chronic exercise training. Interestingly, we showed, for the first time, that the development of DN is closely linked to renal HCD availability. Further research will have to elucidate whether the stimulation of renal HCD levels can be a therapeutic strategy to reduce the risk for developing DN.
AssuntosNefropatias Diabéticas/enzimologia , Dipeptidases/biossíntese , Terapia por Exercício , Glomérulos Renais/enzimologia , Músculo Esquelético/enzimologia , Obesidade/enzimologia , Animais , Nefropatias Diabéticas/genética , Nefropatias Diabéticas/patologia , Dipeptidases/genética , Dipeptídeos/metabolismo , Modelos Animais de Doenças , Indução Enzimática , Histidina/análogos & derivados , Histidina/metabolismo , Humanos , Glomérulos Renais/patologia , Camundongos Transgênicos , Músculo Esquelético/patologia , Obesidade/complicações , Obesidade/genética , Obesidade/patologia , Fatores de Tempo
Histidine is a versatile residue playing key roles in enzyme catalysis thanks to the chemistry of its imidazole group that can serve as nucleophile, general acid or base depending on its protonation state. In bacteria, signal transduction relies on two-component systems (TCS) which comprise a sensor histidine kinase (HK) containing a phosphorylatable catalytic His with phosphotransfer and phosphatase activities over an effector response regulator. Recently, a pH-gated model has been postulated to regulate the phosphatase activity of HisKA HKs based on the pH-dependent rotamer switch of the phosphorylatable His. Here, we have revisited this model from a structural and functional perspective on HK853-RR468 and EnvZ-OmpR TCS, the prototypical HisKA HKs. We have found that the rotamer of His is not influenced by the environmental pH, ruling out a pH-gated model and confirming that the chemistry of the His is responsible for the decrease in the phosphatase activity at acidic pH.
AssuntosHistidina Quinase/química , Histidina Quinase/metabolismo , Thermotoga maritima/enzimologia , Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Catálise , Cristalografia por Raios X , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/metabolismo , Histidina/metabolismo , Histidina Quinase/genética , Concentração de Íons de Hidrogênio , Modelos Biológicos , Modelos Moleculares , Complexos Multienzimáticos/química , Complexos Multienzimáticos/metabolismo , Mutação , Fosforilação , Conformação Proteica , Thermotoga maritima/genética , Transativadores/química , Transativadores/metabolismo
The alpha/beta-Hydrolases (ABH) are a structural class of proteins that are found widespread in nature and includes enzymes that can catalyze various reactions in different substrates. The catalytic versatility of the ABH fold enzymes, which has been a valuable property in protein engineering applications, is based on a similar acid-base-nucleophile catalytic mechanism. In our research, we are concerned with the structure that surrounds the key units of the catalytic machinery, and we have previously found conserved structural organizations that coordinate the catalytic acid, the catalytic nucleophile and the residues of the oxyanion hole. Here, we explore the architecture that surrounds the catalytic histidine at the active sites of enzymes from 40 ABH fold families, where we have identified six conserved interactions that coordinate the catalytic histidine next to the catalytic acid and the catalytic nucleophile. Specifically, the catalytic nucleophile is coordinated next to the catalytic histidine by two weak hydrogen bonds, while the catalytic acid is directly involved in the coordination of the catalytic histidine through by two weak hydrogen bonds. The imidazole ring of the catalytic histidine is coordinated by a CH-π contact and a hydrophobic interaction. Moreover, the catalytic triad residues are connected with a residue that is located at the core of the active site of ABH fold, which is suggested to be the fourth member of a "structural catalytic tetrad". Besides their role in the stability of the catalytic mechanism, the conserved elements of the catalytic site are actively involved in ligand binding and affect other properties of the catalytic activity, such as substrate specificity, enantioselectivity, pH optimum and thermostability of ABH fold enzymes. These properties are regularly targeted in protein engineering applications, and thus, the identified conserved structural elements can serve as potential modification sites in order to develop ABH fold enzymes with altered activities.
AssuntosHistidina/química , Hidrolases/química , Modelos Moleculares , Sítios de Ligação , Catálise , Domínio Catalítico , Histidina/metabolismo , Humanos , Ligação de Hidrogênio , Concentração de Íons de Hidrogênio , Hidrolases/metabolismo , Especificidade por Substrato
Histamine, one of the most toxic and commonly encountered biogenic amines (BA) in food, is produced by the microbial decarboxylation of histidine. It may accumulate at high concentrations in fish and fermented food. Cheese has some of the highest histamine concentrations, the result of the histidine-decarboxylase activity of certain lactic acid bacteria (LAB). The present work describes the nucleotide sequence and transcriptional organization of the gene cluster responsible for histamine biosynthesis (the HDC cluster) in Lactobacillus vaginalis IPLA 11064 isolated from cheese. The influence of histidine availability and pH on histamine production and the expression of the HDC cluster genes is also examined. As expected, the results suggest that the production of histamine under acidic conditions improves cell survival by maintaining the cytosol at an appropriate pH. However, the transcriptional regulation of the HDC cluster is quite different from that described in other dairy histamine-producing LAB, probably due to the lack of a termination-antitermination system in the histidyl-tRNA synthetase gene (hisS).
AssuntosQueijo/microbiologia , Citosol/química , Histamina/biossíntese , Lactobacillus/fisiologia , Animais , Queijo/análise , Regulação Bacteriana da Expressão Gênica , Histidina/análise , Histidina/metabolismo , Histidina Descarboxilase/genética , Concentração de Íons de Hidrogênio , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Lactobacillus/metabolismo , Viabilidade Microbiana
The author has described two new functions of endothelial cells for efficient delivery of drugs to tissues. First, it was indicated that tight junction (TJ)-associated protein, claudin-1, exerts potent paracellular barrier function in cultured mouse lung microvascular endothelial cells (LMECs). This barrier was instantly and reversibly opened by reduction of TJ proteins expression via histamine H1 and H2 receptors. Histamine was biosynthesized by l-histidine decarboxylase from uptaken l-histidine, and biotransformed by type B of monoamine oxidase, suggesting that histamine concentration is controlled in rat brain MECs (BMECs) and LMECs. Moreover, uptake of l-histidine into BMECs and LMECs markedly increased with addition of ZnSO4. Second, it was suggested that drug-metabolizing enzymes such as CYP and flavin-containing monooxygenase exist in vascular endothelial cells exposed to blood and to aerobic conditions. These cells have the same ability to metabolize drugs as hepatocytes, demonstrating that vascular endothelial cells are a metabolic barrier against tissue transfer of drugs. From these results, it was suggested that reversible opening of TJ and selective inhibition of drug metabolism in vascular endothelial cells may be efficient delivery strategies of drugs to tissues. Finally, I hope that this research will lead to development of new drugs and possible re-evaluation of discontinued drugs.
AssuntosSistemas de Liberação de Medicamentos , Células Endoteliais/enzimologia , Células Endoteliais/fisiologia , Animais , Células Cultivadas , Claudina-1/genética , Claudina-1/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Células Endoteliais/metabolismo , Expressão Gênica , Histamina/biossíntese , Histidina/metabolismo , Camundongos , Oxigenases de Função Mista/metabolismo , Ratos , Receptores Histamínicos H1 , Receptores Histamínicos H2
To effective capture and universal enrichment of His-tagged protein, polyacrylic acid (PAA) brushes were used to encapsulate Fe3O4 nanoparticles, connect NTA, and Ni2+ to prepare magnetic beads. These materials provide many advantages, such as excellent stability, tuneable particle size, and a surface for further functionalisation with biomolecules. His-tagged green fluorescence protein (GFP) was separated efficiently, and the binding capacity of Fe3O4/MPS@PAA/NTA-Ni2+ was 93.4â mg/g. Compared with High-Affinity Ni-NTA Resin and Ni-NTA Magnetic Agarose Beads, Fe3O4/MPS@PAA/NTA-Ni2+ nanocomposites exhibited higher separation efficiency and binding capacity towards His-tagged GFP. Moreover, the selectivity and recyclability of them for the target proteins were maintained well after six cycles. This study would widen the application of PAA in constructing multifunctional nanocomposites for biomedical fields.
AssuntosHistidina/metabolismo , Nanopartículas de Magnetita/química , Ácido Nitrilotriacético/análogos & derivados , Compostos Organometálicos/química , Proteínas Recombinantes de Fusão/isolamento & purificação , Resinas Acrílicas/química , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/isolamento & purificação , Histidina/química , Nanocompostos/química , Ácido Nitrilotriacético/química , Ácido Nitrilotriacético/metabolismo , Compostos Organometálicos/metabolismo , Proteínas Recombinantes de Fusão/química
Histidine kinase (HK) of two-component signal transduction system (TCS) is a potential drug target for treating bacterial infections, and most HKs are bifunctional. We have previously identified the HXXXT motif of HK in HisKA subfamily to perform the phosphatase activity, but the specific working mechanism of the threonine is not well understood. In this paper, we use the phosphate group analog BeF3- to capture the enzymatic intermediates between HK853 and RR468 from Thermotoga maritima during dephosphorylation, and demonstrate that the T264 site is essential for populating capable near attack conformers (NAC) between enzyme and substrate to facilitate catalysis. Importantly, mutations at this site can modulate the phosphatase activity of HK. Our results help to understand the TCS signal transduction mechanisms and provide a reference for drug design.
AssuntosHistidina Quinase/metabolismo , Monoéster Fosfórico Hidrolases/metabolismo , Thermotoga maritima/enzimologia , Motivos de Aminoácidos , Histidina/metabolismo , Histidina Quinase/química , Simulação de Dinâmica Molecular , Monoéster Fosfórico Hidrolases/química , Conformação Proteica , Especificidade por Substrato , Thermotoga maritima/química , Thermotoga maritima/metabolismo
Alzheimer's disease (AD) is related to the anomalous binding that occurs between amyloid-ß peptide (Aß) and copper ion, through imidazole ring of histidine (His), as stated in the literature. It is also known that high-affinity metal ion chelators can be pharmacologically used as a possible therapeutic approach. In this work, we tested the ability "in vitro" of chitosan (Chi) to reduce Aß aggregation and Thioflavin T binding assay indicated that chitosan has affinity for Aß and interferes in its aggregation. We also tested the ability of Chi to uptake copper ions in the presence of Aß or His. Equilibrium data reveals that chitosan acted as an effective chelating agent competing with Aß and histidine for copper binding. The addition of histidine or Aß in the system promotes an unfolding of chitosan chains, as verified by small-angle X-ray scattering. Extended X-ray absorption fine structure and XPS spectra show that new copper interactions with groups containing nitrogen in the presence of histidine may occur. These results can help understanding fundamental chemical interactions among species detected in AD and biopolymers, opening up possibilities for new treatment approaches for this disease.
AssuntosPeptídeos beta-Amiloides/metabolismo , Quitosana/metabolismo , Cobre/metabolismo , Histidina/metabolismo , Doença de Alzheimer/metabolismo , Benzotiazóis/química , Biopolímeros/metabolismo , Fluorescência , Humanos
Low-frequency vibrations of l-histidine in a neat solid state and its monohydrochloride monohydrate are both investigated using THz spectroscopy and DFT calculations. The molecular motions in those modes are further quantitatively decomposed into a number of submotions and discussed in terms of their contributions to a mode. The results show significant differences in the averaged contribution percentage of intermolecular motions and the dihedral angle distortions of the imidazole ring between these two crystals. Those phenomena are interpreted from the viewpoint of their hydrogen-bond configurations.
AssuntosHistidina/química , Teoria da Densidade Funcional , Histidina/metabolismo , Modelos Moleculares , Espectroscopia Terahertz/métodos , Termodinâmica
The biosynthesis of histidine, a proteinogenic amino acid, has been extensively studied due to its importance in bacterial growth and survival. Histidinol-phosphate phosphatase (Hol-Pase), which is responsible for the penultimate step of histidine biosynthesis, is generally the last enzyme to be characterized in many bacteria because its origin and evolution are more complex compared to other enzymes in histidine biosynthesis. However, none of the enzymes in histidine biosynthesis, including Hol-Pase, have been characterized in Pseudomonas aeruginosa, which is an important opportunistic Gram-negative pathogen that can cause serious human infections. In our previous work, a transposon mutant of P. aeruginosa was found to display a growth defect on glucose-containing minimal solid medium. In this study, we found that the growth defect was due to incomplete histidine auxotrophy caused by PA0335 inactivation. Subsequently, PA0335 was shown to encode Hol-Pase, and its function and enzymatic activity were investigated using genetic and biochemical methods. In addition to PA0335, the roles of 12 other predicted genes involved in histidine biosynthesis in P. aeruginosa were examined. Among them, hisC2 (PA3165), hisH2 (PA3152), and hisF2 (PA3151) were found to be dispensable for histidine synthesis, whereas hisG (PA4449), hisE (PA5067), hisF1 (PA5140), hisB (PA5143), hisI (PA5066), hisC1 (PA4447), and hisA (PA5141) were essential because deletion of each resulted in complete histidine auxotrophy; similar to the case for PA0335, hisH1 (PA5142) or hisD (PA4448) deletion caused incomplete histidine auxotrophy. Taken together, our results outline the histidine synthesis pathway of P. aeruginosa IMPORTANCE Histidine is a common amino acid in proteins. Because it plays critical roles in bacterial metabolism, its biosynthetic pathway in many bacteria has been elucidated. However, the pathway remains unclear in Pseudomonas aeruginosa, an important opportunistic pathogen in clinical settings; in particular, there is scant knowledge about histidinol-phosphate phosphatase (Hol-Pase), which has a complex origin and evolution. In this study, P. aeruginosa Hol-Pase was identified and characterized. Furthermore, the roles of all other predicted genes involved in histidine biosynthesis were examined. Our results illustrate the histidine synthesis pathway of P. aeruginosa The knowledge obtained from this study may help in developing strategies to control P. aeruginosa-related infections. In addition, some enzymes of the histidine synthesis pathway from P. aeruginosa might be used as elements of histidine synthetic biology in other industrial microorganisms.
AssuntosProteínas de Bactérias/genética , Histidina/metabolismo , Histidinol-Fosfatase/genética , Pseudomonas aeruginosa/genética , Proteínas de Bactérias/metabolismo , Vias Biossintéticas , Histidinol-Fosfatase/metabolismo , Pseudomonas aeruginosa/enzimologia , Pseudomonas aeruginosa/metabolismo
The natural resistance-associated macrophage protein (Nramp) family encompasses transition metal and proton cotransporters that are present in many organisms from bacteria to humans. Recent structures of Deinococcus radiodurans Nramp (DraNramp) in multiple conformations revealed the intramolecular rearrangements required for alternating access of the metal-binding site to the external or cytosolic environment. Here, using recombinant proteins and metal transport and cysteine accessibility assays, we demonstrate that two parallel cytoplasm-accessible networks of conserved hydrophilic residues in DraNramp, one lining the wide intracellular vestibule for metal release and the other forming a narrow proton transport pathway, are essential for metal transport. We further show that mutagenic or posttranslational modifications of transmembrane helix (TM) 6b, which structurally links these two pathways, impede normal conformational cycling and metal transport. TM6b contains two highly conserved histidines, His232 and His237 We found that different mutagenic perturbations of His232, just below the metal-binding site along the proton exit route, differentially affect DraNramp's conformational state, suggesting that His232 serves as a pivot point for conformational changes. In contrast, any replacement of His237, lining the metal exit route, locked the transporter in a transport-inactive outward-closed state. We conclude that these two histidines, and TM6b more broadly, help trigger the bulk rearrangement of DraNramp to the inward-open state upon metal binding and facilitate return of the empty transporter to an outward-open state upon metal release.
AssuntosProteínas de Transporte de Cátions/química , Deinococcus/química , Histidina/química , Metais/metabolismo , Sequência de Aminoácidos/genética , Proteínas de Transporte de Cátions/genética , Proteínas de Transporte de Cátions/metabolismo , Cobalto/química , Cobalto/metabolismo , Deinococcus/genética , Deinococcus/metabolismo , Histidina/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Transporte de Íons , Manganês/química , Manganês/metabolismo , Metais/química , Modelos Moleculares , Mutação , Conformação Proteica , Processamento de Proteína Pós-Traducional/genética , Prótons
Liquid-liquid phase separation (LLPS) of intrinsically disordered proteins (IDPs) is involved in both intracellular membraneless organelles and extracellular tissues. Despite growing understanding of LLPS, molecular-level mechanisms behind this process are still not fully established. Here, we use histidine-rich squid beak proteins (HBPs) as model IDPs to shed light on molecular interactions governing LLPS. We show that LLPS of HBPs is mediated though specific modular repeats. The morphology of separated phases (liquid-like versus hydrogels) correlates with the repeats' hydrophobicity. Solution-state NMR indicates that LLPS is a multistep process initiated by deprotonation of histidine residues, followed by transient hydrogen bonding with tyrosine, and eventually by hydrophobic interactions. The microdroplets are stabilized by aromatic clustering of tyrosine residues exhibiting restricted molecular mobility in the nano-to-microsecond timescale according to solid-state NMR experiments. Our findings provide guidelines to rationally design pH-responsive peptides with LLPS ability for various applications, including bioinspired protocells and smart drug-delivery systems.
AssuntosBico/metabolismo , Decapodiformes/metabolismo , Histidina/metabolismo , Proteínas Intrinsicamente Desordenadas/metabolismo , Tirosina/metabolismo , Animais , Bico/química , Materiais Biocompatíveis , Biopolímeros/química , Biopolímeros/metabolismo , Coloides/química , Coloides/metabolismo , Ligação de Hidrogênio , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Proteínas Intrinsicamente Desordenadas/química , Espectroscopia de Ressonância Magnética , Microscopia , Engenharia de Proteínas/métodos , Espalhamento a Baixo Ângulo
BACKGROUND: Histamine is a key mediator of the anti-inflammatory activity conferred by the probiotic organism Lactobacillus reuteri ATCC PTA 6475 in animal models of colitis and colorectal cancer. In L. reuteri, histamine synthesis and secretion requires L-histidine decarboxylase and a L-histidine/histamine exchanger. Chloride channel (ClC)-family proton/chloride antiporters have been proposed to act as electrochemical shunts in conjunction with amino acid decarboxylase systems, correcting ion imbalances generated by decarboxylation through fixed ratio exchange of two chloride ions for one proton. This family is unique among transporters by facilitating ion flux in either direction. Here we examine the histidine decarboxylase system in relation to ClC antiporters in the probiotic organism Lactobacillus reuteri. RESULTS: In silico analyses reveal that L. reuteri possesses two ClC transporters, EriC and EriC2, as well as a complete histidine decarboxylase gene cluster (HDC) for the synthesis and export of histamine. When the transport activity of either proton/chloride antiporter is disrupted by genetic manipulation, bacterial histamine output is reduced. Using fluorescent reporter assays, we further show that ClC transporters affect histamine output by altering intracellular pH and membrane potential. ClC transport also alters the expression and activity of two key HDC genes: the histidine decarboxylase (hdcA) and the histidine/histamine exchanger (hdcP). CONCLUSIONS: Histamine production is a potentially beneficial feature for intestinal microbes by promoting long-term colonization and suppression of inflammation and host immune responses. ClC transporters may serve as tunable modulators for histamine production by L. reuteri and other gut microbes.
AssuntosCanais de Cloreto/metabolismo , Histidina/metabolismo , Lactobacillus reuteri/metabolismo , Transporte Biológico , Concentração de Íons de Hidrogênio , Potenciais da Membrana
The unique capability of proton buffering is the rationale for using histidine (HIS) as a component of solutions for induction of cardiac arrest and myocardial protection in cardiac surgery. In humans, infusion of cardioplegic solution may increase blood plasma HIS from ~ 70 to ~ 21,000 µM. We examined the effects of a large intravenous dose of HIS on ammonia and amino acid concentrations and energy status of the body. Rats received 198 mM HIS intravenously (20 ml/kg) or vehicle. Samples of blood plasma, urine, liver, and soleus (SOL) and extensor digitorum longus (EDL) muscles were analysed at 2 or 24 h after treatment. At 2 h after HIS load, we found higher HIS concentration in all examined tissues, higher urea and ammonia concentrations in blood and urine, lower ATP content and higher AMP/ATP ratio in the liver and muscles, higher concentrations of almost all examined amino acids in urine, and lower glycine concentration in blood plasma, liver, and muscles when compared with controls. Changes in other amino acids were tissue dependent, markedly increased alanine and glutamate in the blood and the liver. At 24 h, the main findings were lower ATP concentrations in muscles, lower concentrations of branched-chain amino acids (BCAA; valine, leucine, and isoleucine) in blood plasma and muscles, and higher carnosine content in SOL when compared with controls. It is concluded that a load of large HIS dose results in increased ammonia levels and marked alterations in amino acid and energy metabolism. Pathogenesis is discussed in the article.


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Nollywood Gists
- Oct- 2020 -20 October
Yvonne Jegede, Sharon Ooja, Charles Inojie comfort Ada Ameh as she Mourns the Sad Passing of Her Daughter
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- 17 October
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In 2019, Nigeria's first-ever entry for the international feature film Oscar category, "Lionheart", the directorial debut movie of Genevieve Nnaji was struck out of the Oscars as it predominantly features…Continue reading »
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78 & Thriving! We Can't Get Over these Photos of Idowu Philips a.k.a Iya Rainbow
It's Idowu Philips, also known as Iya Rainbow's birthday and we can't believe she's 78. So vibrant, happy and full of life… 78 looks super cool on her. She definitely makes…Continue reading »
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Follow the Story of Mr & Mrs Williams on Ayope Davies' Latest Short Film "Marriage Is Easy"
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- 12 October
The Official Trailer for Dimeji Ajibola's Anticipated Movie "Ratnik" Is Here
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- 10 October
Teniola Olatoni's "The New Normal" will be in Cinemas November 20th | Watch the Trailer
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Watch the Season Finale of "Witches" by Yemisi Wada
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#BNMovieFeature: You Don't Want to Miss Niyi Akinmolayan's Short Film "Room 315"
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The Official Trailer for Kunle Afolayan's "Citation" is Here
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The Fragile Bond between a Young Married Couple is Tested in "Heaven Baby" starring Oreka Godis, Ibrahim Suleiman | Watch
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Now releasing for Conflict of the Eagles is the ultra-realistic Libyan Civil War mod, created on a whole new hand-made and highly detailed map of Libya. Fight as democratic rebel fighters, Islamists, or Ghaddafi's Jamahiriya forces and fight for military superiority as Libya devolves into a revolution of separate militias.
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Refusing to go to the gym counts as resistance training, right?
I'm not sure if I lost my camo pants or if they're just doing a really good job.
They tell you that you'll lose your mind when you grow older. What they don't tell you is that you won't miss it very much.
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I saw my dad chopping up onions today and I cried. Onions was a good dog.
It is very easy to become a superman, you just have to change the sequence of clothes while wearing.


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Computer Science > Machine Learning
Title: ARSM Gradient Estimator for Supervised Learning to Rank
(Submitted on 1 Nov 2019 (v1), last revised 18 Feb 2020 (this version, v2))
Abstract: We propose a new model for supervised learning to rank. In our model, the relevance labels are assumed to follow a categorical distribution whose probabilities are constructed based on a scoring function. We optimize the training objective with respect to the multivariate categorical variables with an unbiased and low-variance gradient estimator. Learning-to-rank methods can generally be categorized into pointwise, pairwise, and listwise approaches. Although our scoring function is pointwise, the proposed framework permits flexibility over the choice of the loss function. In our new model, the loss function need not be differentiable and can either be pointwise or listwise. Our proposed method achieves better or comparable results on two datasets compared with existing pairwise and listwise methods.
Submission historyFrom: Shahin Boluki [view email]
[v1] Fri, 1 Nov 2019 17:05:29 GMT (21kb)
[v2] Tue, 18 Feb 2020 05:36:29 GMT (16kb)
Link back to: arXiv, form interface, contact.


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No. 4-S-33, Kota Bharu City Point, Jalan Ismail Hilir No. 4-S-33, Kota Bharu City Point, Jalan Ismail Hilir, Kota Bharu 15000, Malaysia Show Map
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Archaicawful White Kitchen Cart With Granite Top Ideas Ceramic As Marble Fabric
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If you want to get in on the trend of using kitchen carts in your kitchen, you need to know about all the variety of kitchen carts that are on the market. No longer are you limited to a cart with a top and one shelf underneath? Those old-fashioned kitchen carts have been revamped in a number of styles and designs that will fit your specific entertaining needs. Whether it is in the kitchen, or as a serving assistant, there are a wide variety of kitchen carts out there.
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On a popular home network, a couple that was doing a remodel while living in the home used a cart. While installing granite countertops. They used this makeshift cart to keep their cooking items. This was a great solution that allowed them to keep living comfortably while remodeling.
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In the kitchens of yesterday, the kitchen table served as the center of activity in the kitchen. This is where people prepared the evening meal, socialized, and dined.
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