The Miller–Urey experiment

From Wikipedia, the free encyclopedia

https://en.wikipedia.org/wiki/Miller%E2%80%93Urey_experiment

 

The experiment

The Miller–Urey experiment (or Miller experiment) was a chemical experiment that simulated the conditions thought at the time (1952) to be present on the early Earth and tested the chemical origin of life under those conditions. The experiment at the time supported Alexander Oparin's and J. B. S. Haldane's hypothesis that putative conditions on the primitive Earth favoured chemical reactions that synthesized more complex organic compounds from simpler inorganic precursors. Considered to be the classic experiment investigating abiogenesis, it was performed in 1952 by Stanley Miller, supervised by Harold Urey at the University of Chicago, and published the following year.

After Miller's death in 2007, scientists examining sealed vials preserved from the original experiments were able to show that there were actually well over 20 different amino acids produced in Miller's original experiments. That is considerably more than what Miller originally reported, and more than the 20 that naturally occur in the genetic code. More recent evidence suggests that Earth's original atmosphere might have had a composition different from the gas used in the Miller experiment, but prebiotic experiments continue to produce racemic mixtures of simple-to-complex compounds under varying conditions.

Experiment

Play media

Descriptive video of the experiment

The experiment used water (H₂O), methane (CH₄), ammonia (NH₃), and hydrogen (H₂). The chemicals were all sealed inside a sterile 5-liter glass flask connected to a 500 ml flask half-full of water. The water in the smaller flask was heated to induce evaporation, and the water vapour was allowed to enter the larger flask. Continuous electrical sparks were fired between the electrodes to simulate lightning in the water vapour and gaseous mixture, and then the simulated atmosphere was cooled again so that the water condensed and trickled into a U-shaped trap at the bottom of the apparatus.

After a day, the solution collected at the trap had turned pink in colour, and after a week of continuous operation the solution was deep red and turbid. The boiling flask was then removed, and mercuric chloride was added to prevent microbial contamination. The reaction was stopped by adding barium hydroxide and sulfuric acid, and evaporated to remove impurities. Using paper chromatography, Miller identified five amino acids present in the solution: glycine, α-alanine and β-alanine were positively identified, while aspartic acid and α-aminobutyric acid (AABA) were less certain, due to the spots being faint.

In a 1996 interview, Stanley Miller recollected his lifelong experiments following his original work and stated: "Just turning on the spark in a basic pre-biotic experiment will yield 11 out of 20 amino acids."

The original experiment remained in 2017 under the care of Miller and Urey's former student Jeffrey Bada, a professor at the UCSD, Scripps Institution of Oceanography. As of 2013, the apparatus used to conduct the experiment was on display at the Denver Museum of Nature and Science.

Chemistry of experiment

One-step reactions among the mixture components can produce hydrogen cyanide (HCN), formaldehyde (CH₂O), and other active intermediate compounds (acetylene, cyanoacetylene, etc.):

CO₂ → CO + [O] (atomic oxygen)

CH₄ + 2[O] → CH₂O + H₂O

CO + NH₃ → HCN + H₂O

CH₄ + NH₃ → HCN + 3H₂ (BMA process)

The formaldehyde, ammonia, and HCN then react by Strecker synthesis to form amino acids and other biomolecules:

CH₂O + HCN + NH₃ → NH₂-CH₂-CN + H₂O

NH₂-CH₂-CN + 2H₂O → NH₃ + NH₂-CH₂-COOH (glycine)

Furthermore, water and formaldehyde can react, via Butlerov's reaction to produce various sugars like ribose.

The experiments showed that simple organic compounds of building blocks of proteins and other macromolecules can be formed from gases with the addition of energy.

Other experiments

This experiment inspired many others. In 1961, Joan Oró found that the nucleotide base adenine could be made from hydrogen cyanide (HCN) and ammonia in a water solution. His experiment produced a large amount of adenine, the molecules of which were formed from 5 molecules of HCN. Also, many amino acids are formed from HCN and ammonia under these conditions. Experiments conducted later showed that the other RNA and DNA nucleobases could be obtained through simulated prebiotic chemistry with a reducing atmosphere.

There also had been similar electric discharge experiments related to the origin of life contemporaneous with Miller–Urey. An article in The New York Times (March 8, 1953:E9), titled "Looking Back Two Billion Years" describes the work of Wollman (William) M. MacNevin at The Ohio State University, before the Miller Science paper was published in May 1953. MacNevin was passing 100,000 volt sparks through methane and water vapor and produced "resinous solids" that were "too complex for analysis." The article describes other early earth experiments being done by MacNevin. It is not clear if he ever published any of these results in the primary scientific literature.

K. A. Wilde submitted a paper to Science on December 15, 1952, before Miller submitted his paper to the same journal on February 10, 1953. Wilde's paper was published on July 10, 1953. Wilde used voltages up to only 600 V on a binary mixture of carbon dioxide (CO₂) and water in a flow system. He observed only small amounts of carbon dioxide reduction to carbon monoxide, and no other significant reduction products or newly formed carbon compounds. Other researchers were studying UV-photolysis of water vapor with carbon monoxide. They have found that various alcohols, aldehydes and organic acids were synthesized in reaction mixture.

More recent experiments by chemists Jeffrey Bada, one of Miller's graduate students, and Jim Cleaves at Scripps Institution of Oceanography of the University of California, San Diego were similar to those performed by Miller. However, Bada noted that in current models of early Earth conditions, carbon dioxide and nitrogen (N₂) create nitrites, which destroy amino acids as fast as they form. When Bada performed the Miller-type experiment with the addition of iron and carbonate minerals, the products were rich in amino acids. This suggests the origin of significant amounts of amino acids may have occurred on Earth even with an atmosphere containing carbon dioxide and nitrogen.

Earth's early atmosphere

Some evidence suggests that Earth's original atmosphere might have contained fewer of the reducing molecules than was thought at the time of the Miller–Urey experiment. There is abundant evidence of major volcanic eruptions 4 billion years ago, which would have released carbon dioxide, nitrogen, hydrogen sulfide (H₂S), and sulfur dioxide (SO₂) into the atmosphere. Experiments using these gases in addition to the ones in the original Miller–Urey experiment have produced more diverse molecules. The experiment created a mixture that was racemic (containing both L and D enantiomers) and experiments since have shown that "in the lab the two versions are equally likely to appear"; however, in nature, L amino acids dominate. Later experiments have confirmed disproportionate amounts of L or D oriented enantiomers are possible.

Originally it was thought that the primitive secondary atmosphere contained mostly ammonia and methane. However, it is likely that most of the atmospheric carbon was CO₂ with perhaps some CO and the nitrogen mostly N₂. In practice gas mixtures containing CO, CO₂, N₂, etc. give much the same products as those containing CH₄ and NH₃ so long as there is no O₂. The hydrogen atoms come mostly from water vapor. In fact, in order to generate aromatic amino acids under primitive earth conditions it is necessary to use less hydrogen-rich gaseous mixtures. Most of the natural amino acids, hydroxyacids, purines, pyrimidines, and sugars have been made in variants of the Miller experiment.

More recent results may question these conclusions. The University of Waterloo and University of Colorado conducted simulations in 2005 that indicated that the early atmosphere of Earth could have contained up to 40 percent hydrogen—implying a much more hospitable environment for the formation of prebiotic organic molecules. The escape of hydrogen from Earth's atmosphere into space may have occurred at only one percent of the rate previously believed based on revised estimates of the upper atmosphere's temperature. One of the authors, Owen Toon notes: "In this new scenario, organics can be produced efficiently in the early atmosphere, leading us back to the organic-rich soup-in-the-ocean concept... I think this study makes the experiments by Miller and others relevant again." Outgassing calculations using a chondritic model for the early earth complement the Waterloo/Colorado results in re-establishing the importance of the Miller–Urey experiment.

In contrast to the general notion of early earth's reducing atmosphere, researchers at the Rensselaer Polytechnic Institute in New York reported the possibility of oxygen available around 4.3 billion years ago. Their study reported in 2011 on the assessment of Hadean zircons from the earth's interior (magma) indicated the presence of oxygen traces similar to modern-day lavas. This study suggests that oxygen could have been released in the earth's atmosphere earlier than generally believed.

Extraterrestrial sources

Conditions similar to those of the Miller–Urey experiments are present in other regions of the solar system, often substituting ultraviolet light for lightning as the energy source for chemical reactions. The Murchison meteorite that fell near Murchison, Victoria, Australia in 1969 was found to contain over 90 different amino acids, nineteen of which are found in Earth life. Comets and other icy outer-solar-system bodies are thought to contain large amounts of complex carbon compounds (such as tholins) formed by these processes, darkening surfaces of these bodies. The early Earth was bombarded heavily by comets, possibly providing a large supply of complex organic molecules along with the water and other volatiles they contributed. This has been used to infer an origin of life outside of Earth: the panspermia hypothesis.

Recent related studies

In recent years, studies have been made of the amino acid composition of the products of "old" areas in "old" genes, defined as those that are found to be common to organisms from several widely separated species, assumed to share only the last universal ancestor (LUA) of all extant species. These studies found that the products of these areas are enriched in those amino acids that are also most readily produced in the Miller–Urey experiment. This suggests that the original genetic code was based on a smaller number of amino acids – only those available in prebiotic nature – than the current one.

Jeffrey Bada, himself Miller's student, inherited the original equipment from the experiment when Miller died in 2007. Based on sealed vials from the original experiment, scientists have been able to show that although successful, Miller was never able to find out, with the equipment available to him, the full extent of the experiment's success. Later researchers have been able to isolate even more different amino acids, 25 altogether. Bada has estimated that more accurate measurements could easily bring out 30 or 40 more amino acids in very low concentrations, but the researchers have since discontinued the testing. Miller's experiment was therefore a remarkable success at synthesizing complex organic molecules from simpler chemicals, considering that all known life uses just 20 different amino acids.

In 2008, a group of scientists examined 11 vials left over from Miller's experiments of the early 1950s. In addition to the classic experiment, reminiscent of Charles Darwin's envisioned "warm little pond", Miller had also performed more experiments, including one with conditions similar to those of volcanic eruptions. This experiment had a nozzle spraying a jet of steam at the spark discharge. By using high-performance liquid chromatography and mass spectrometry, the group found more organic molecules than Miller had. They found that the volcano-like experiment had produced the most organic molecules, 22 amino acids, 5 amines and many hydroxylated molecules, which could have been formed by hydroxyl radicals produced by the electrified steam. The group suggested that volcanic island systems became rich in organic molecules in this way, and that the presence of carbonyl sulfide there could have helped these molecules form peptides.

The main problem of theories based around amino acids is the difficulty in obtaining spontaneous formation of peptides. Since John Desmond Bernal's suggestion that clay surfaces could have played a role in abiogenesis, scientific efforts have been dedicated to investigating clay-mediated peptide bond formation, with limited success. Peptides formed remained over-protected and shown no evidence of inheritance or metabolism. In December 2017 a theoretical model developed by Erastova and collaborators suggested that peptides could form at the interlayers of layered double hydroxides such as green rust in early earth conditions. According to the model, drying of the intercalated layered material should provide energy and co-alignment required for peptide bond formation in a ribosome-like fashion, while re-wetting should allow mobilising the newly formed peptides and repopulate the interlayer with new amino acids. This mechanism is expected to lead to the formation of 12+ amino acid-long peptides within 15-20 washes. Researches also observed slightly different adsorption preferences for different amino acids, and postulated that, if coupled to a diluted solution of mixed amino acids, such preferences could lead to sequencing.

In October 2018, researchers at McMaster University on behalf of the Origins Institute announced the development of a new technology, called a Planet Simulator, to help study the origin of life on planet Earth and beyond.

Amino acids identified

Below is a table of amino acids produced and identified in the "classic" 1952 experiment, as published by Miller in 1953, the 2008 re-analysis of vials from the volcanic spark discharge experiment, and the 2010 re-analysis of vials from the H₂S-rich spark discharge experiment.

Amino acid	Produced in experiment			Proteinogenic
	Miller–Urey (1952)	Volcanic spark discharge (2008)	H2S-rich spark discharge (2010)
Glycine				Yes
α-Alanine				Yes
β-Alanine				No
Aspartic acid				Yes
α-Aminobutyric acid				No
Serine				Yes
Isoserine				No
α-Aminoisobutyric acid				No
β-Aminoisobutyric acid				No
β-Aminobutyric acid				No
γ-Aminobutyric acid				No
Valine				Yes
Isovaline				No
Glutamic acid				Yes
Norvaline				No
α-Aminoadipic acid				No
Homoserine				No
2-Methylserine				No
β-Hydroxyaspartic acid				No
Ornithine				No
2-Methylglutamic acid				No
Phenylalanine				Yes
Homocysteic acid				No
S-Methylcysteine				No
Methionine				Yes
Methionine sulfoxide				No
Methionine sulfone				No
Isoleucine				Yes
Leucine				Yes
Ethionine				No
Cysteine				Yes
Histidine				Yes
Lysine				Yes
Asparagine				Yes
Pyrrolysine				Yes
Proline				Yes
Glutamine				Yes
Arginine				Yes
Threonine				Yes
Selenocysteine				Yes
Tryptophan				Yes
Tyrosine				Yes

Urea cycle

From Wikipedia, the free encyclopedia

The urea cycle (also known as the ornithine cycle) is a cycle of biochemical reactions that produces urea (NH₂)₂CO from ammonia (NH₃). This cycle occurs in ureotelic organisms. The urea cycle converts highly toxic ammonia to urea for excretion. This cycle was the first metabolic cycle to be discovered (Hans Krebs and Kurt Henseleit, 1932), five years before the discovery of the TCA cycle. This cycle was described in more detail later on by Ratner and Cohen. The urea cycle takes place primarily in the liver and, to a lesser extent, in the kidneys.

Function

Amino acid catabolism results in waste ammonia. All animals need a way to excrete this product. Most aquatic organisms, or ammonotelic organisms, excrete ammonia without converting it. Organisms that cannot easily and safely remove nitrogen as ammonia convert it to a less toxic substance such as urea via the urea cycle, which occurs mainly in the liver. Urea produced by the liver is then released into the bloodstream where it travels to the kidneys and is ultimately excreted in urine. The urea cycle is essential to these organisms, because if the nitrogen or ammonia are not eliminated from the organism it can be very detrimental. In species including birds and most insects, the ammonia is converted into uric acid or its urate salt, which is excreted in solid form.

Reactions

The entire process converts two amino groups, one from NH⁺
₄ and one from Aspartate, and a carbon atom from HCO⁻
₃, to the relatively nontoxic excretion product urea at the cost of four "high-energy" phosphate bonds (3 ATP hydrolyzed to 2 ADP and one AMP). The conversion from ammonia to urea happens in five main steps. The first is needed for ammonia to enter the cycle and the following four are all a part of the cycle itself. To enter the cycle, ammonia is converted to carbamoyl phosphate. The urea cycle consists of four enzymatic reactions: one mitochondrial and three cytosolic.

Reactions of the urea cycle

Step	Reactants	Products	Catalyzed by	Location
1	NH3 + HCO− 3 + 2ATP	carbamoyl phosphate + 2ADP + Pi	CPS1	mitochondria
2	carbamoyl phosphate + ornithine	citrulline + Pi	OTC, zinc, biotin	mitochondria
3	citrulline + aspartate + ATP	argininosuccinate + AMP + PPi	ASS	cytosol
4	argininosuccinate	arginine + fumarate	ASL	cytosol
5	arginine + H2O	ornithine + urea	ARG1, manganese	cytosol

The reactions of the urea cycle

1 L-ornithine
2 carbamoyl phosphate
3 L-citrulline
4 argininosuccinate
5 fumarate
6 L-arginine
7 urea
L-Asp L-aspartate
CPS-1 carbamoyl phosphate synthetase I
OTC Ornithine transcarbamoylase
ASS argininosuccinate synthetase
ASL argininosuccinate lyase
ARG1 arginase 1

First reaction: entering the urea cycle

Before the urea cycle begins ammonia is converted to carbamoyl phosphate. The reaction is catalyzed by carbamoyl phosphate synthetase I and requires the use of two ATP molecules. The carbamoyl phosphate then enters the urea cycle.

Steps of the urea cycle

1. Carbamoyl phosphate is converted to citrulline. With catalysis by ornithine transcarbamoylase, the carbamoyl phosphate group is donated to ornithine and releases a phosphate group.
2. A condensation reaction occurs between the amino group of aspartate and the carbonyl group of citrulline to form argininosuccinate. This reaction is ATP dependent and is catalyzed by argininosuccinate synthetase.
3. Argininosuccinate undergoes cleavage by argininosuccinase to form arginine and fumarate.
4. Arginine is cleaved by arginase to form urea and ornithine. The ornithine is then transported back to the mitochondria to begin the urea cycle again.

Overall reaction equation

In the first reaction, NH⁺
₄ + HCO⁻
₃ is equivalent to NH₃ + CO₂ + H₂O.

Thus, the overall equation of the urea cycle is:

• NH₃ + CO₂ + aspartate + 3 ATP + 3 H₂O → urea + fumarate + 2 ADP + 2 P_i + AMP + PP_i + H₂O

Since fumarate is obtained by removing NH₃ from aspartate (by means of reactions 3 and 4), and PP_i + H₂O → 2 P_i, the equation can be simplified as follows:

• 2 NH₃ + CO₂ + 3 ATP + 3 H₂O → urea + 2 ADP + 4 P_i + AMP

Note that reactions related to the urea cycle also cause the production of 2 NADH, so the overall reaction releases slightly more energy than it consumes. The NADH is produced in two ways:

• One NADH molecule is produced by the enzyme glutamate dehydrogenase in the conversion of glutamate to ammonium and α-ketoglutarate. Glutamate is the non-toxic carrier of amine groups. This provides the ammonium ion used in the initial synthesis of carbamoyl phosphate.
• The fumarate released in the cytosol is hydrated to malate by cytosolic fumarase. This malate is then oxidized to oxaloacetate by cytosolic malate dehydrogenase, generating a reduced NADH in the cytosol. Oxaloacetate is one of the keto acids preferred by transaminases, and so will be recycled to aspartate, maintaining the flow of nitrogen into the urea cycle.

We can summarize this by combining the reactions:

• CO₂ + glutamate + aspartate + 3 ATP + 2 NAD⁺+ 3 H₂O → urea + α-ketoglutarate + oxaloacetate + 2 ADP + 2 P_i + AMP + PP_i + 2 NADH

The two NADH produced can provide energy for the formation of 5 ATP (cytosolic NADH provides 2.5 ATP with the malate-aspartate shuttle in human liver cell), a net production of two high-energy phosphate bond for the urea cycle. However, if gluconeogenesis is underway in the cytosol, the latter reducing equivalent is used to drive the reversal of the GAPDH step instead of generating ATP.

The fate of oxaloacetate is either to produce aspartate via transamination or to be converted to phosphoenolpyruvate, which is a substrate for gluconeogenesis.

Products of the urea cycle

As stated above many vertebrates use the urea cycle to create urea out of ammonium so that the ammonium does not damage the body. Though this is helpful, there are other effects of the urea cycle. For example: consumption of two ATP, production of urea, generation of H+, the combining of HCO3- and NH4+ to forms where it can be regenerated, and finally the consumption of NH4+.

Regulation

N-Acetylglutamic acid

The synthesis of carbamoyl phosphate and the urea cycle are dependent on the presence of N-acetylglutamic acid (NAcGlu), which allosterically activates CPS1. NAcGlu is an obligate activator of carbamoyl phosphate synthetase. Synthesis of NAcGlu by N-acetylglutamate synthase (NAGS) is stimulated by both Arg, allosteric stimulator of NAGS, and Glu, a product in the transamination reactions and one of NAGS's substrates, both of which are elevated when free amino acids are elevated. So Glu not only is a substrate for NAGS but also serves as an activator for the urea cycle.

Substrate concentrations

The remaining enzymes of the cycle are controlled by the concentrations of their substrates. Thus, inherited deficiencies in cycle enzymes other than ARG1 do not result in significant decreases in urea production (if any cycle enzyme is entirely missing, death occurs shortly after birth). Rather, the deficient enzyme's substrate builds up, increasing the rate of the deficient reaction to normal.

The anomalous substrate buildup is not without cost, however. The substrate concentrations become elevated all the way back up the cycle to NH⁺
₄, resulting in hyperammonemia (elevated [NH⁺
₄]_P).

Although the root cause of NH⁺
₄ toxicity is not completely understood, a high [NH⁺
₄] puts an enormous strain on the NH⁺
₄-clearing system, especially in the brain (symptoms of urea cycle enzyme deficiencies include intellectual disability and lethargy). This clearing system involves GLUD1 and GLUL, which decrease the 2-oxoglutarate (2OG) and Glu pools. The brain is most sensitive to the depletion of these pools. Depletion of 2OG decreases the rate of TCAC, whereas Glu is both a neurotransmitter and a precursor to GABA, another neurotransmitter. 

Link with the citric acid cycle

The urea cycle and the citric acid cycle are independent cycles but are linked. One of the nitrogens in the urea cycle is obtained from the transamination of oxaloacetate to aspartate. The fumarate that is produced in step three is also an intermediate in the citric acid cycle and is returned to that cycle.

Urea cycle disorders

Urea cycle disorders are rare and affect about one in 35,000 people in the United States. Genetic defects in the enzymes involved in the cycle can occur, which usually manifest within a few days after birth. The recently born child will typically experience varying bouts of emesis and periods of lethargy.

Ultimately the infant may go into a coma and develop brain damage. Newborns with UCD are at a much higher risk of complications or death due to untimely screening tests and misdiagnosed cases. The most common misdiagnosis is neonatal sepsis. Signs of UCD can be present within the first 2-3 days of life, however, the present method to get confirmation by test results can take too long and can potentially cause complications such as coma or death.

Urea cycle disorders may also be diagnosed in adults, and symptoms may include delirium episodes, lethargy, and symptoms similar to that of a stroke. On top of these symptoms if the urea cycle begins to malfunction in the liver the patient may obtain cirrhosis which can also lead to sarcopenia (the loss of muscle mass). Mutations lead to deficiencies of the various enzymes and transporters involved in the urea cycle and cause urea cycle disorders. If individuals with a defect in any of the six enzymes used in the cycle ingest amino acids beyond what is necessary for the minimum daily requirements, then the ammonia that is produced will not be able to be converted to urea. These individuals can experience hyperammonemia or the buildup of a cycle intermediate.

Individual disorders

• N-Acetylglutamate synthase (NAGS) deficiency
• Carbamoyl phosphate synthetase (CPS) deficiency
• Ornithine transcarbamoylase (OTC) deficiency
• Citrullinemia Type I (Deficiency of argininosuccinic acid synthase)
• Argininosuccinic aciduria (Deficiency of argininosuccinic acid lyase)
• Argininemia (Deficiency of arginase)
• Hyperornithinemia, hyperammonemia, homocitrullinuria (HHH) syndrome (Deficiency of the mitochondrial ornithine transporter)

All urea cycle defects, except OTC deficiency, are inherited in an autosomal recessive manner. OTC deficiency is inherited as an X-linked recessive disorder, although some females can show symptoms. Most urea cycle disorders are associated with hyperammonemia, however argininemia and some forms of argininosuccinic aciduria do not present with elevated ammonia.

 

Le Chatelier's principle

From Wikipedia, the free encyclopedia

Jump to navigation Jump to search

Le Chatelier's principle (pronounced UK: /lə ʃæˈtɛljeɪ/ or US: /ˈʃɑːtəljeɪ/), also called Chatelier's principle or "The Equilibrium Law", is a principle of chemistry used to predict the effect of a change in conditions on chemical equilibria. The principle is named after French chemist Henry Louis Le Chatelier, and sometimes also credited to Karl Ferdinand Braun, who discovered it independently. It can be stated as:

When any system at equilibrium for a long period of time is subjected to a change in concentration, temperature, volume, or pressure, (1) the system changes to a new equilibrium, and (2) this change partly counteracts the applied change.

It is common to treat the principle as a more general observation of systems, such as:

When a settled system is disturbed, it will adjust to diminish the change that has been made to it:

or, "roughly stated",

Any change in status quo prompts an opposing reaction in the responding system.

The concept of systemic maintenance of an equilibrium state despite perturbations has a variety of names, depending upon the discipline using it (e.g. homeostasis, an idea which encompasses the concept, is commonly used in biology), and has been studied in a variety of contexts, chiefly in the natural sciences. In chemistry, the principle is used to manipulate the outcomes of reversible reactions, often to increase their yield. In pharmacology, the binding of ligands to receptors may shift the equilibrium according to Le Chatelier's principle, thereby explaining the diverse phenomena of receptor activation and desensitization. In economics, the principle has been generalized to help explain the price equilibrium of efficient economic systems.

Phenomena in apparent contradiction to Le Chatelier's principle can also arise in systems of simultaneous equilibrium (see response reactions).

As a physical law

Le Chatelier's principle describes the qualitative behavior of systems where there is an externally induced, instantaneous change in one parameter of a system; it states that a behavioral shift occurs in the system so as to oppose (partly cancel) the parameter change. The duration of adjustment depends on the strength of the negative feedback to the initial shock. Where a shock initially induces positive feedback (such as thermal runaway), the new equilibrium can be far from the old one, and can take a long time to reach. In some dynamic systems, the end-state cannot be determined from the shock. The principle is typically used to describe closed negative-feedback systems, but applies, in general, to thermodynamically closed and isolated systems in nature, since the second law of thermodynamics ensures that the disequilibrium caused by an instantaneous shock must have a finite half-life. The principle has analogs throughout the entire physical world.

While well rooted in chemical equilibrium and extended into economic theory, Le Chatelier's principle can also be used in describing mechanical systems in that a system put under stress will respond in such a way as to reduce or minimize that stress. Moreover, the response will generally be via the mechanism that most easily relieves that stress. Shear pins and other such sacrificial devices are design elements that protect systems against stress applied in undesired manners to relieve it so as to prevent more extensive damage to the entire system, a practical engineering application of Le Chatelier's principle.

Chemistry

Effect of change in concentration

Changing the concentration of a chemical will shift the equilibrium to the side that would counter that change in concentration. The chemical system will attempt to partly oppose the change affected to the original state of equilibrium. In turn, the rate of reaction, extent, and yield of products will be altered corresponding to the impact on the system.

This can be illustrated by the equilibrium of carbon monoxide and hydrogen gas, reacting to form methanol.

CO + 2 H₂ ⇌ CH₃OH

Suppose we were to increase the concentration of CO in the system. Using Le Chatelier's principle, we can predict that the amount of methanol will increase, decreasing the total change in CO. If we are to add a species to the overall reaction, the reaction will favor the side opposing the addition of the species. Likewise, the subtraction of a species would cause the reaction to "fill the gap" and favor the side where the species was reduced. This observation is supported by the collision theory. As the concentration of CO is increased, the frequency of successful collisions of that reactant would increase also, allowing for an increase in forward reaction, and generation of the product. Even if the desired product is not thermodynamically favored, the end-product can be obtained if it is continuously removed from the solution.

The effect of a change in concentration is often exploited synthetically for condensation reactions (i.e., reactions that extrude water) that are equilibrium processes (e.g., formation of an ester from carboxylic acid and alcohol or an imine from an amine and aldehyde). This can be achieved by physically sequestering water, by adding desiccants like anhydrous magnesium sulfate or molecular sieves, or by continuous removal of water by distillation, often facilitated by a Dean-Stark apparatus.

Effect of change in temperature

The reversible reaction N₂O₄(g) ⇌ 2NO₂(g) is endothermic, so the equilibrium position can be shifted by changing the temperature.
When heat is added and the temperature increases, the reaction shifts to the right and the flask turns reddish brown due to an increase in NO₂. This demonstrates Le Chatelier's principle: the equilibrium shifts in the direction that consumes energy.
When heat is removed and the temperature decreases, the reaction shifts to the left and the flask turns colorless due to an increase in N₂O₄: again, according to Le Chatelier's principle.

The effect of changing the temperature in the equilibrium can be made clear by 1) incorporating heat as either a reactant or a product, and 2) assuming that an increase in temperature increases the heat content of a system. When the reaction is exothermic (ΔH is negative and energy is released), heat is included as a product, and when the reaction is endothermic (ΔH is positive and energy is consumed), heat is included as a reactant. Hence, whether increasing or decreasing the temperature would favor the forward or the reverse reaction can be determined by applying the same principle as with concentration changes.

Take, for example, the reversible reaction of nitrogen gas with hydrogen gas to form ammonia:

N₂(g) + 3 H₂(g) ⇌ 2 NH₃(g)    ΔH = -92 kJ mol⁻¹

Because this reaction is exothermic, it produces heat:

N₂(g) + 3 H₂(g) ⇌ 2 NH₃(g) + heat

If the temperature were increased, the heat content of the system would increase, so the system would consume some of that heat by shifting the equilibrium to the left, thereby producing less ammonia. More ammonia would be produced if the reaction were run at a lower temperature, but a lower temperature also lowers the rate of the process, so, in practice (the Haber process) the temperature is set at a compromise value that allows ammonia to be made at a reasonable rate with an equilibrium concentration that is not too unfavorable.

In exothermic reactions, an increase in temperature decreases the equilibrium constant, K, whereas in endothermic reactions, an increase in temperature increases K.

Le Chatelier's principle applied to changes in concentration or pressure can be understood by giving K a constant value. The effect of temperature on equilibria, however, involves a change in the equilibrium constant. The dependence of K on temperature is determined by the sign of ΔH. The theoretical basis of this dependence is given by the Van 't Hoff equation.

Effect of change in pressure

The equilibrium concentrations of the products and reactants do not directly depend on the total pressure of the system. They may depend on the partial pressures of the products and reactants, but if the number of moles of gaseous reactants is equal to the number of moles of gaseous products, pressure has no effect on equilibrium.

Changing total pressure by adding an inert gas at constant volume does not affect the equilibrium concentrations (see Effect of adding an inert gas below).

Changing total pressure by changing the volume of the system changes the partial pressures of the products and reactants and can affect the equilibrium concentrations (see §Effect of change in volume below).

Effect of change in volume

Changing the volume of the system changes the partial pressures of the products and reactants and can affect the equilibrium concentrations. With a pressure increase due to a decrease in volume, the side of the equilibrium with fewer moles is more favorable and with a pressure decrease due to an increase in volume, the side with more moles is more favorable. There is no effect on a reaction where the number of moles of gas is the same on each side of the chemical equation.

Considering the reaction of nitrogen gas with hydrogen gas to form ammonia:

N₂ + 3 H₂4 moles ⇌ 2 NH₃2 moles    ΔH = -92kJ mol⁻¹

Note the number of moles of gas on the left-hand side and the number of moles of gas on the right-hand side. When the volume of the system is changed, the partial pressures of the gases change. If we were to decrease pressure by increasing volume, the equilibrium of the above reaction will shift to the left, because the reactant side has a greater number of moles than does the product side. The system tries to counteract the decrease in partial pressure of gas molecules by shifting to the side that exerts greater pressure. Similarly, if we were to increase pressure by decreasing volume, the equilibrium shifts to the right, counteracting the pressure increase by shifting to the side with fewer moles of gas that exert less pressure. If the volume is increased because there are more moles of gas on the reactant side, this change is more significant in the denominator of the equilibrium constant expression, causing a shift in equilibrium.

Effect of adding an inert gas

An inert gas (or noble gas), such as helium, is one that does not react with other elements or compounds. Adding an inert gas into a gas-phase equilibrium at constant volume does not result in a shift. This is because the addition of a non-reactive gas does not change the equilibrium equation, as the inert gas appears on both sides of the chemical reaction equation. For example, if A and B react to form C and D, but X does not participate in the reaction: ${\displaystyle {\ce {{\mathit {a}}A{}+{\mathit {b}}B{}+{\mathit {x}}X<=>{\mathit {c}}C{}+{\mathit {d}}D{}+{\mathit {x}}X}}}$. While it is true that the total pressure of the system increases, the total pressure does not have any effect on the equilibrium constant; rather, it is a change in partial pressures that will cause a shift in the equilibrium. If, however, the volume is allowed to increase in the process, the partial pressures of all gases would be decreased resulting in a shift towards the side with the greater number of moles of gas. The shift will never occur on the side with fewer moles of gas. It is also known as Le Chatelier's postulate.

Effect of a catalyst

A catalyst increases the rate of a reaction without being consumed in the reaction. The use of a catalyst does not affect the position and composition of the equilibrium of a reaction, because both the forward and backward reactions are sped up by the same factor.

For example, consider the Haber process for the synthesis of ammonia (NH₃):

N₂ + 3 H₂ ⇌ 2 NH₃

In the above reaction, iron (Fe) and molybdenum (Mo) will function as catalysts if present. They will accelerate any reactions, but they do not affect the state of the equilibrium.

General statement of Le Chatelier's principle

Le Chatelier's principle refers to states of thermodynamic equilibrium. The latter are stable against perturbations that satisfy certain criteria; this is essential to the definition of thermodynamic equilibrium.

For this, a state of thermodynamic equilibrium is most conveniently described through a fundamental relation that specifies a cardinal function of state, of the energy kind, or of the entropy kind, as a function of state variables chosen to fit the thermodynamic operations through which a perturbation is to be applied.

In theory and, nearly, in some practical scenarios, a body can be in a stationary state with zero macroscopic flows and rates of chemical reaction (for example, when no suitable catalyst is present), yet not in thermodynamic equilibrium, because it is metastable or unstable; then Le Chatelier's principle does not necessarily apply.

General statements related to Le Chatelier's principle

A body can also be in a stationary state with non-zero rates of flow and chemical reaction; sometimes the word "equilibrium" is used in reference to such states, though by definition they are not thermodynamic equilibria. Sometimes, it is proposed to consider Le Chatelier's principle for such states. For this exercise, rates of flow and of chemical reaction must be considered. Such rates are not supplied by equilibrium thermodynamics. For such states, it has turned out to be difficult or unfeasible to make valid and very general statements that echo Le Chatelier's principle. Prigogine and Defay demonstrate that such a scenario may or may not exhibit moderation, depending upon exactly what conditions are imposed after the perturbation.

Economics

In economics, a similar concept also named after Le Chatelier was introduced by American economist Paul Samuelson in 1947. There the generalized Le Chatelier principle is for a maximum condition of economic equilibrium: Where all unknowns of a function are independently variable, auxiliary constraints—"just-binding" in leaving initial equilibrium unchanged—reduce the response to a parameter change. Thus, factor-demand and commodity-supply elasticities are hypothesized to be lower in the short run than in the long run because of the fixed-cost constraint in the short run.

Since the change of the value of an objective function in a neighbourhood of the maximum position is described by the envelope theorem, Le Chatelier's principle can be shown to be a corollary thereof.

Frostbite

From Wikipedia, the free encyclopedia

 

Frostbite
Other names	Frostnip
Frostbitten toes two to three days after mountain climbing
Specialty	Dermatology Emergency medicine, orthopedics
Symptoms	Numbness, feeling cold, clumsy, pale color
Complications	Hypothermia, compartment syndrome
Types	Superficial, deep
Causes	Temperatures below freezing
Risk factors	Alcohol, smoking, mental health problems, certain medications, prior cold injury
Diagnostic method	Based on symptoms
Differential diagnosis	Frostnip, pernio, trench foot
Prevention	Avoid cold, wear proper clothing, maintain hydration and nutrition, stay active without becoming exhausted
Treatment	Rewarming, medication, surgery
Medication	Ibuprofen, tetanus vaccine, iloprost, thrombolytics
Frequency	Unknown

Frostbite occurs when exposure to low temperatures causes freezing of the skin or other tissues. The initial symptom is typically numbness. This may be followed by clumsiness with a white or bluish color to the skin. Swelling or blistering may occur following treatment. The hands, feet, and face are most commonly affected. Complications may include hypothermia or compartment syndrome.

People who are exposed to low temperatures for prolonged periods, such as winter sports enthusiasts, military personnel, and homeless individuals, are at greatest risk. Other risk factors include drinking alcohol, smoking, mental health problems, certain medications, and prior injuries due to cold. The underlying mechanism involves injury from ice crystals and blood clots in small blood vessels following thawing. Diagnosis is based on symptoms. Severity may be divided into superficial (1st and 2nd degree) or deep (3rd and 4th degree). A bone scan or MRI may help in determining the extent of injury.

Prevention is through wearing proper clothing, maintaining hydration and nutrition, avoiding low temperatures, and staying active without becoming exhausted. Treatment is by rewarming. This should be done only when refreezing is not a concern. Rubbing or applying snow to the affected part is not recommended. The use of ibuprofen and tetanus toxoid is typically recommended. For severe injuries iloprost or thrombolytics may be used. Surgery is sometimes necessary. Amputation, however, should generally be delayed for a few months to allow determination of the extent of injury.

The number of cases of frostbite is unknown. Rates may be as high as 40% a year among those who mountaineer. The most common age group affected is those 30 to 50 years old. Evidence of frostbite occurring in people dates back 5,000 years. Frostbite has also played an important role in a number of military conflicts. The first formal description of the condition was in 1813 by Dominique Jean Larrey, a physician in Napoleon's army, during its invasion of Russia.

Signs and symptoms

Frostbite

Areas that are usually affected include cheeks, ears, nose and fingers and toes. Frostbite is often preceded by frostnip. The symptoms of frostbite progress with prolonged exposure to cold. Historically, frostbite has been classified by degrees according to skin and sensation changes, similar to burn classifications. However, the degrees do not correspond to the amount of long term damage. A simplification of this system of classification is superficial (first or second degree) or deep injury (third or fourth degree).

First degree

• First degree frostbite is superficial, surface skin damage that is usually not permanent.
• Early on, the primary symptom is loss of feeling in the skin. In the affected areas, the skin is numb, and possibly swollen, with a reddened border.
• In the weeks after injury, the skin's surface may slough off.

Second degree

• In second degree frostbite, the skin develops clear blisters early on, and the skin's surface hardens.
• In the weeks after injury, this hardened, blistered skin dries, blackens, and peels.
• At this stage, lasting cold sensitivity and numbness can develop.

Third degree

• In third degree frostbite, the layers of tissue below the skin freeze.
• Symptoms include blood blisters and "blue-grey discoloration of the skin".
• In the weeks after injury, pain persists and a blackened crust (eschar) develops.
• There can be longterm ulceration and damage to growth plates.

Fourth degree

Frostbite 12 days later

• In fourth degree frostbite, structures below the skin are involved like muscles, tendon, and bone.
• Early symptoms include a colorless appearance of the skin, a hard texture, and painless rewarming.
• Later, the skin becomes black and mummified. The amount of permanent damage can take one month or more to determine. Autoamputation can occur after two months.

Causes

Risk factors

The major risk factor for frostbite is exposure to cold through geography, occupation and/or recreation. Inadequate clothing and shelter are major risk factors. Frostbite is more likely when the body's ability to produce or retain heat is impaired. Physical, behavioral, and environmental factors can all contribute to the development of frostbite. Immobility and physical stress (such as malnutrition or dehydration) are also risk factors. Disorders and substances that impair circulation contribute, including diabetes, Raynaud's phenomenon, tobacco and alcohol use. Homeless individuals and individuals with some mental illnesses may be at higher risk.

Mechanism

Freezing

In frostbite, cooling of the body causes narrowing of the blood vessels (vasoconstriction). Temperatures below −4 °C (25 °F) are required to form ice crystals in the tissues. The process of freezing causes ice crystals to form in the tissue, which in turn causes damage at the cellular level. Ice crystals can damage cell membranes directly. In addition, ice crystals can damage small blood vessels at the site of injury. Scar tissue forms when fibroblasts replace the dead cells.

Rewarming

Rewarming causes tissue damage through reperfusion injury, which involves vasodilation, swelling (edema), and poor blood flow (stasis). Platelet aggregation is another possible mechanism of injury. Blisters and spasm of blood vessels (vasospasm) can develop after rewarming.

Non-freezing cold injury

The process of frostbite differs from the process of non-freezing cold injury (NFCI). In NFCI, temperature in the tissue decreases gradually. This slower temperature decrease allows the body to try to compensate through alternating cycles of closing and opening blood vessels (vasoconstriction and vasodilation). If this process continues, inflammatory mast cells act in the area. Small clots (microthrombi) form and can cut off blood to the affected area (known as ischemia) and damage nerve fibers. Rewarming causes a series of inflammatory chemicals such as prostaglandins to increase localized clotting.

Pathophysiology

The pathological mechanism by which frostbite causes body tissue injury can be characterized by four stages: Prefreeze, freeze-thaw, vascular stasis, and the late ischemic stage.

1. Prefreeze phase: involves the cooling of tissues without ice crystal formation.
2. Freeze-thaw phase: ice-crystals form, resulting in cellular damage and death.
3. Vascular stasis phase: marked by blood coagulation or the leaking of blood out of the vessels.
4. Late ischemic phase: characterized by inflammatory events, ischemia and tissue death.

Diagnosis

Frostbite is diagnosed based on signs and symptoms as described above, and by patient history. Other conditions that can have a similar appearance or occur at the same time include:

• Frostnip is similar to frostbite, but without ice crystal formation in the skin. Whitening of the skin and numbness reverse quickly after rewarming.
• Trench foot is damage to nerves and blood vessels that results exposure to wet, cold (non-freezing) conditions. This is reversible if treated early.
• Pernio or chilblains are inflammation of the skin from exposure to wet, cold (non-freezing) conditions. They can appear as various types of ulcers and blisters.
• Bullous pemphigoid is a condition that causes itchy blisters over the body that can mimic frostbite. It does not require exposure to cold to develop.
• Levamisole toxicity is a vasculitis that can appear similar to frostbite. It is caused by contamination of cocaine by levamisole. Skin lesions can look similar those of frostbite, but do not require cold exposure to occur.

People who have hypothermia often have frostbite as well. Since hypothermia is life-threatening this should be treated first. Technetium-99 or MR scans are not required for diagnosis, but might be useful for prognostic purposes.

Prevention

The Wilderness Medical Society recommends covering the skin and scalp, taking in adequate nutrition, avoiding constrictive footwear and clothing, and remaining active without causing exhaustion. Supplemental oxygen might also be of use at high elevations. Repeated exposure to cold water makes people more susceptible to frostbite. Additional measures to prevent frostbite include:

• Avoiding temperatures below −15 °C (5 °F)
• Avoiding moisture, including in the form of sweat and/or skin emollients
• Avoiding alcohol and drugs that impair circulation or natural protective responses
• Layering clothing
• Using chemical or electric warming devices
• Recognizing early signs of frostnip and frostbite

Treatment

Individuals with frostbite or potential frostbite should go to a protected environment and get warm fluids. If there is no risk of re-freezing, the extremity can be exposed and warmed in the groin or underarm of a companion. If the area is allowed to refreeze, there can be worse tissue damage. If the area cannot be reliably kept warm, the person should be brought to a medical facility without rewarming the area. Rubbing the affected area can also increase tissue damage. Aspirin and ibuprofen can be given in the field to prevent clotting and inflammation. Ibuprofen is often preferred to aspirin because aspirin may block a subset of prostaglandins that are important in injury repair.

The first priority in people with frostbite should be to assess for hypothermia and other life-threatening complications of cold exposure. Before treating frostbite, the core temperature should be raised above 35 °C. Oral or intravenous (IV) fluids should be given.

Other considerations for standard hospital management include:

• wound care: blisters can be drained by needle aspiration, unless they are bloody (hemorrhagic). Aloe vera gel can be applied before breathable, protective dressings or bandages are put on.
• antibiotics: if there is trauma, skin infection (cellulitis) or severe injury
• tetanus toxoid: should be administered according to local guidelines. Uncomplicated frostbite wounds are not known to encourage tetanus.
• pain control: NSAIDs or opioids are recommended during the painful rewarming process.

Rewarming

If the area is still partially or fully frozen, it should be rewarmed in the hospital with a warm bath with povidone iodine or chlorhexidine antiseptic. Active rewarming seeks to warm the injured tissue as quickly as possible without burning. The faster tissue is thawed, the less tissue damage occurs.

According to Handford and colleagues, "The Wilderness Medical Society and State of Alaska Cold Injury Guidelines recommend a temperature of 37–39 °C, which decreases the pain experienced by the patient whilst only slightly slowing rewarming time." Warming takes 15 minutes to 1 hour. Rewarming can be very painful, so pain management is important.

Medications

People with potential for large amputations and who present within 24 hours of injury can be given TPA with heparin. These medications should be withheld if there are any contraindications. Bone scans or CT angiography can be done to assess damage.

Blood vessel dilating medications such as iloprost may prevent blood vessel blockage. This treatment might be appropriate in grades 2–4 frostbite, when people get treatment within 48 hours. In addition to vasodilators, sympatholytic drugs can be used to counteract the detrimental peripheral vasoconstriction that occurs during frostbite.

Surgery

Various types of surgery might be indicated in frostbite injury, depending on the type and extent of damage. Debridement or amputation of necrotic tissue is usually delayed unless there is gangrene or systemic infection (sepsis). This has led to the adage "Frozen in January, amputate in July". If symptoms of compartment syndrome develop, fasciotomy can be done to attempt to preserve blood flow.

Prognosis

3 weeks after initial frostbite

Tissue loss and autoamputation are potential consequences of frostbite. Permanent nerve damage including loss of feeling can occur. It can take several weeks to know what parts of the tissue will survive. Time of exposure to cold is more predictive of lasting injury than temperature the individual was exposed to. The classification system of grades, based on the tissue response to initial rewarming and other factors is designed to predict degree of longterm recovery.

Grades

Grade 1: if there is no initial lesion on the area, no amputation or lasting effects are expected

Grade 2: if there is a lesion on the distal body part, tissue and fingernails can be destroyed

Grade 3: if there is a lesion on the intermediate or near body part, autoamputation and loss of function can occur

Grade 4: if there is a lesion very near the body (such as the carpals of the hand), the limb can be lost. Sepsis and/or other systemic problems are expected.

A number of long term sequelae can occur after frostbite. These include transient or permanent changes in sensation, paresthesia, increased sweating, cancers, and bone destruction/arthritis in the area affected.

Epidemiology

There is a lack of comprehensive statistics about the epidemiology of frostbite. In the United States, frostbite is more common in northern states. In Finland, annual incidence was 2.5 per 100,000 among civilians, compared with 3.2 per 100,000 in Montreal. Research suggests that men aged 30–49 are at highest risk, possibly due to occupational or recreational exposures to cold.

History

Frostbite has been described in military history for millennia. The Greeks encountered and discussed the problem of frostbite as early as 400 BCE. Researchers have found evidence of frostbite in humans dating back 5,000 years, in an Andean mummy. Napoleon's Army was the first documented instance of mass cold injury in the early 1800s. According to Zafren, nearly 1 million combatants fell victim to frostbite in the First and Second World Wars, and the Korean War.

Society and culture

Mountaineer Nigel Vardy in hospital after suffering frostbite when benighted on Denali in 1999. His nose, fingers and toes were subsequently amputated.

Several notable cases of frostbite include: Captain Lawrence Oates, an English army captain and Antarctic explorer who in 1912 died of complications of frostbite; noted American rock climber Hugh Herr, who in 1982 lost both legs below the knee to frostbite after being stranded on Mount Washington (New Hampshire) in a blizzard; Beck Weathers, a survivor of the 1996 Mount Everest disaster who lost his nose and hands to frostbite; Scottish mountaineer Jamie Andrew, who in 1999 had all four limbs amputated due to sepsis from frostbite sustained after becoming trapped for four nights whilst climbing Les Droites in the Mont Blanc massif.

Research directions

Evidence is insufficient to determine whether or not hyperbaric oxygen therapy as an adjunctive treatment can assist in tissue salvage. Cases have been reported, but no randomized control trial has been performed on humans.

Medical sympathectomy using intravenous reserpine has also been attempted with limited success.

 Studies have suggested that administration of tissue plasminogen activator (tPa) either intravenously or intra-arterially may decrease the likelihood of eventual need for amputation.