Oxidative Stress and aging

Oxidative stress is the major pillar of the theory of aging. As a joke, I often say in my classes that we get old because we have the bad habit of spending our whole life breathing oxygen. Basically it is the oxygen that makes us live, but it is also the one that kills us little by little, that is, that makes us grow old...
And what is the relationship between oxygen and aging? The answer boils down to two words: oxidative stress! Sporadically, there are O2 molecules that transform into reactive oxygen species, most of which are neutralized by our antioxidant defenses (more information on this subject here). However, there are always some reactive oxygen species that can bypass our defenses and consequently can cause minor damage to some of our biomolecules. Although these damages do not have much biological significance, when evaluated isolated, as we grow older, they accumulate, and these cumulative damages begin to translate to the loss of some functionalities. Examples are loss of skin malleability, joint stiffness, loss of sensory ability, etc.

Therefore, everything that can accelerate our metabolic rate has the potential to make us age faster because it increases the production of reactive oxygen species. In this context, the effect of emotional stress is particularly evident! For example, people who have jobs and activities of high stress, age at a much higher rate than those who have a much more relaxed life.
Finally, I would like to make it clear that oxidative stress is not the only factor responsible for aging, but it is certainly one of the main ones, so if we want to age more slowly, we have to ensure an adequate balance between the pro-oxidants and the anti-oxidants!

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Metabolic map about LDL and atherosclerosis

Myoglobin

Myoglobin is a cytoplasmic hemoprotein composed by a single polypeptide chain of 154 amino acids. It is expressed solely in cardiac myocytes and oxidative skeletal muscle fibers. Myoglobin was so named because of its functional and structural similarity to hemoglobin. Like hemoglobin, myoglobin binds reversibly to O2 and thus may facilitate the transport of O2 from red blood cells to the mitochondria during periods of increased metabolic activity or serve as an O 2 reservoir during hypoxia or anoxia.The structure of myoglobin was first delineated by John Kendrew more than 40 years ago and subsequent work has shown that it is a polypeptide chain consisting of eight α-helices. It binds oxygen to its heme residue, a porphyrin ring with an iron ion. The polypeptide chain is folded and packs the heme prosthetic group, positioning it between two histidine, His64 and His93 residues. The iron ion interacts with six ligands, four of which are supplied by the nitrogen atoms of the four pyrrhols and share a common plane. The side chain imidazole of His93, provides the fifth ligand, stabilizing the heme group and slightly displacing the iron ion out of the heme plane. The position of the sixth ligand, in deoximoglobin, serves as the binding site for O2, as well as for other potential ligands, such as CO or NO. When O2 binds, the iron ion, it is partially drawn back toward the porphyrin plane. Although this shift is of little importance in the function of monomeric myoglobin, it provides the basis for the conformational changes that underlie the allosteric properties of tetrameric hemoglobin. In addition, studies using X-ray diffraction and xenon binding techniques have identified four highly conserved internal cavities within the myoglobin molecule that can help target molecules to bind to the heme residue.Related to its role as an O2 reservoir, myoglobin also functions as an intracellular pO2 buffer (partial pressure of O2). Similarly to the role of creatine phosphokinase, which works to buffer ATP concentrations when muscle activity increases, myoglobin works to buffer O2 concentrations. As a result, the intracellular concentration of O2 remains relatively constant and homogeneous, despite increases in O2 flow from the capillaries to the mitochondria, induced by physical activity.

Text written by:

Ana Rita Cardoso

João Faria

Joel Mateus

Pedro Desport
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Cartoon about Cell-fies

Oxidative stress and cellular respiration

During cellular respiration, electrons are transferred from NADH or FADH2, along 4 protein complexes in the inner mitochondrial membrane, to an O₂ molecule (read more about this subject here). In the last stage of the process, the electrons are transported one by one, that is, they will reach the oxygen one at a time. 

This situation, which may seem only a detail to many, has, in fact, very important implications for our biochemistry, because it means that all O₂ molecules are, even temporarily, transformed into a free radical, the superoxide anion. This means that, literally, at every instant we are producing large quantities of reactive oxygen species. However, this situation, which is potentially very dangerous, does not have, under normal conditions, dramatic consequences for cells, mainly for 2 reasons:
1. There are mechanisms that prevent the superoxide anion from diffusing from complex 4 before it is completely reduced to water. That is, the free radical is formed, but remains in place and quickly receives another electron, ceasing to be free radical.
2. As there are always some superoxide anions that can escape the first mechanism, we have other defense mechanisms, and in this context, the most important is the presence of a mitochondrial enzyme called superoxide dismutase. This enzyme, which also has a cytosolic isoform, will cause dismutation of the superoxide anion, converting two of these molecules into hydrogen peroxide.
Of course there will also be superoxide anions that will be able to escape from superoxide dismutase, but under normal conditions these are very few. In addition, we still have several other antioxidant defenses waiting for them...

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Metabolic map about the surface proteins of cytotoxic T cells

Insulin

Insulin is a polypeptide hormone produced, stored and secreted in Beta cells of the islets of Langerhans, in the pancreas (in a histological section it is seen that they occupy the central part). It is an anabolic hormone that acts at the level of the liver, adipose tissue and with influence in the brain.
This protein has two polypeptide chains, with 21 amino acids in the A chain and 30 in the B chain, joined by disulfide bonds, which gives a greater stability and a correct folding. It begins to be produced in the form of pre-pro-insulin which, by action of the peptidase is cleaved to form the proinsulin. The proteolytic cleavage of peptide C forms the two chain bioactive insulin, which is stored in secretory granules for subsequent insulin secretion.Its main function is to regulate blood glucose levels in a context of hyperglycemia. In this way, glucose acts as a biochemical signal that triggers its secretion. Thus, when carbohydrate-containing foods are absorbed, glucose is metabolized to ATP and this in turn triggers insulin secretion. Protein-protein interactions and phosphorylations are used to transmit the signal. In adipose tissue and muscle, the binding of insulin to membrane receptors triggers the displacement of GLUT4-rich vesicles that fuse with the membrane, increasing cell uptake, being an insulin-dependent transport.
On the other hand, in the liver, insulin activates the enzyme glycokinase, which is responsible for the conversion of glucose into glucose-6-phosphate; Guarantees an intracellular concentration of glucose lower than the extracellular concentration and, therefore, a gradient of glucose concentration favorable to its entry into these cells, through the GLUT-2 transporter, following metabolization by glycolysis, Krebs and the respiratory chain to produce ATP. Thus, after food intake, glucose is absorbed into the intestines and is released into the bloodstream, causing blood concentrations to rise, leading to transient hyperglycemia. The pancreas releases insulin to lower glucose concentration, allowing glucose to be consumed by the cells, as well as stimulating the storage of glucose in the liver in the form of glycogen; The liver also metabolize glucose into triacylglycerols, transported as VLDL to be stored in adipose tissue, which are useful reserves in fasting situations. Signal transmission ceases, at meal time, by dephosphorylation of the insulin receptor by protein tyrosine phosphatase.
In summary, insulin stimulates glycogenesis, fatty acid synthesis and glycolysis and inhibits antagonistic pathways: glycogenolysis, fatty acid degradation and hepatic gluconeogenesis. It also stimulates protein synthesis. It has action on inherent enzymes as well as effects on gene transcription. It also acts on specific receptors in the hypothalamus to inhibit the act of eating, thus regulating feeding and energy conservation.
Inborn errors of beta cell metabolism can produce excessive or defective production of insulin by gene mutations (GCK), Kir 6.2 alterations, or insulin synthesis transcription factors, respectively. Increased glucose leads to increased osmotic pressure, glycation of proteins and formation of reactive oxygen species (EROS).
Diabetes is the metabolic disease characterized by increased blood sugar: It may be Type I - in which the body stops producing insulin by destroying the B cells of the pancreas. It is important to check for symptoms of polydipsia, fruity aroma breathing, blood glucose and blood ketones levels. Essential therapies focus on insulin therapy, fluid replacement, replacement of electrolytes and nourishment. In turn, in Type II diabetes, the cells do not produce enough insulin to lower the concentration of gucose or there is a condition of insulin resistance. Adipocytes, myocytes and hepatocytes do not respond correctly. It presents symptoms similar to type I but more gradual. It is necessary to test for fasting blood glucose and for abnormal levels to continue the investigation for glycemic curve; glycated hemoglobin, control alcohol consumption, etc.
They can lead to complications such as diabetic retinopathy, atherosclerosis, diabetic nephropathy, neuropathy, myocardial infarction/stroke, infections - leucocytes less effective in hyperglycemia, hypertension and oxidation of blood vessels. There are currently several drugs on the market that address problems with insulin, as well as different types of injectable insulin depending on the cause of the disease and the purpose of action.

Text written by:

Denilson Araújo

Prescília Sampa

Solange da Costa

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Cartoon sobre milagre científico

Oxidative stress - Advantages and disadvantages

Oxidative stress results primarily from an imbalance between molecules potentially dangerous to our cells, the so-called reactive oxygen species, and molecules that protect the oxidative integrity of our cellular structures, as discussed in another post (more information here). When this imbalance favors the former, or disadvantages the latter, we have the condition called oxidative stress.
Oxidative stress is the mainstay of the aging theory, because although we have several antioxidant defenses to protect us, there are always reactive oxygen species that can bypass these defenses, causing little damages that start to accumulate. Furthermore, in the case of smokers, there is permanent oxidative stress, especially at the level of lung cells, since tobacco smoke contains large amounts of reactive oxygen species (and reactive nitrogen species, but I will not talk about them today), which causes the antioxidant defenses in the lungs to be unable to cope completely with the aggressions from tobacco smoke.
But not everything is bad news, because our biochemistry is full of examples where even the most dangerous situations/molecules can be converted into an advantage, at least in some contexts... This is what happens with oxidative stress! Although it is a potentially fatal situation for cells and therefore, most often, is a situation we should avoid, there is a context where oxidative stress is beneficial to our body. I'm talking about the inflammatory response...

In a simple way, when there is an invading microorganism (or other types of stimuli), our organism detects that something is not well, and initiates the inflammatory response. One of the most important cellular components of it is neutrophils, a class of white blood cells. One of the ways neutrophils act, is related to their contact with invading microorganisms. In response to this situation, neutrophils increase their metabolic rate, and the reason is simple: they want to overproduce reactive oxygen species, that means, they want to induce oxidative stress. Of course, this is a controlled process, that is, the stimulation of oxidative stress occurs at a level that can still be effectively eliminated by our antioxidant defenses, but most microorganisms will no longer have this capability. Thus, neutrophils induce oxidative stress, at a level still tolerated by most of our cells, but not tolerated by most microorganisms. In this way, the invasion is controlled and ideally does not cause significant damage to our body.
Therefore, even oxidative stress can be advantageous, as long as properly controlled. It is another notable example of how fascinating is the World of Biochemistry ... ;)

Metabolic map about propionyl-CoA utilization

Hemoglobin

For higher animals, simple diffusion mechanisms in body fluids are not an efficient way to meet the oxygenation needs of their tissues and cellular material. To the low area/volume ratio of these living beings, it is added the fact that O2 is a molecule that is essentially insoluble, which makes its transport even more difficult. The solution then passes through carrier proteins, associated with erythrocytes - hemoglobin, to which the following lines refer.Hemoglobin is an oligomeric protein and is generally a metalloprotein consisting of about 600 amino acids, arranged in 2 alpha chains and 2 paired beta chains in a quaternary globular structure. The four chains constitute the organic part of the molecule, and are attached to heme prosthetic groups (consisting of a porphyrin ring and a transition metal: Fe2+) which have affinity for the O2 molecules because of the electron configuration. It is the Fe2+ that assumes this function, always in its ferrous form, and the ferric form - Fe3+ - is not able to bind O2, being at the same time more unstable and prone to the formation of reactive species. Fe2+ has one O2 binding site and this bond as expected would be reversible to allow oxygen to be transported to where it is needed. Due to this binding, there is a change of color in human blood, from bright red when it is in its oxygenated form, to a more purplish tone in its venous phase. Some molecules such as CO2 and NO have a higher affinity for the heme group, "expelling" O2 molecules from erythrocytes, which explains their toxicity to the organism. 

Porphyrias are genetic diseases related to porphyrin of the heme group. Examples are acute intermittent porphyria and accumulation of uroporphyrogen I each with specific symptoms.
Concerning the coordinated transport of O2, CO2 and H+, the mechanism is as follows: 

O2 binds cooperatively to hemoglobin (this means that the bonds promote more bonds) and then the affinity of hemoglobin varies with pH. In an acidic environment, H+ and CO2 cause the release of O2 whereas in a basic medium, O2 causes the release of H+ and CO2. This is the so-called Bohr effect(reciprocal effect): CO2 + H2O <-> HCO3- + H+
The dead erythrocytes release the heme group generating: Fe3+ (which is recycled) and bilirubin (which is excreted in the liver). The latter may have a negative effect if released into the blood because it causes jaundice, or a positive antioxidant effect especially as an antioxidant of the membrane, because it collects two hydroperoxide radicals, having about 1/10 the efficiency of vitamin C.

Text written by:
Beatriz Ribeiro
Cláudia Campos
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Cartoon about meiosis

Oxidative stress - Antioxidants

Recently I have made a post about oxidative stress (you can read it here), in which, of course, I gave some prominence to the reactive oxygen species. Well, today I'm going to talk about the "good ones", that is, the antioxidants...
The word "antioxidant" is probably the word most often heard in social media ads, whether in the context of food, cosmetics, etc. And, in fact, we can (and should!) ensure a high exogenous supply of antioxidants, being this an important issue in different contexts. What possibly fewer people know is that we already have several internal antioxidants. Therefore, we can already divide the antioxidants into 2 categories:
- Exogenous antioxidants, which are those that we obtain mainly from the diet;
- Endogenous antioxidants, which are those that we produce in our cells and that, under normal conditions, are always present in them.
Another possible classification is as follows:
- Enzymatic antioxidants, which are enzymes that we produce and whose function is to eliminate reactive oxygen species. For example, there is an enzyme, called superoxide dismutase that catalyzes the conversion of 2 superoxide anions (that are free radicals), to a hydrogen peroxide molecule (which, although being a reactive oxygen species, is not a free radical). Another example is catalase (you can read more about thisenzyme here), which converts hydrogen peroxide into two products potentially harmless to our biomolecules, water and oxygen.
- Non-enzymatic antioxidants, which are molecules that function as antioxidants because they react with reactive oxygen species, promoting their inactivation. In the background, they are molecules that "generously" put themselves at the forefront of the battle against the pro-oxidants. Therefore, these pro-oxidants will react with them, promoting their oxidation. This situation is beneficial, because it is the antioxidants that end up getting oxidized, sparing our biomolecules from oxidative damage. These non-enzymatic antioxidants often have in their composition benzene rings which stabilize the presence of a possible unpaired electron, and may also react with one another so that their unpaired electrons become paired. 

Within this class we have glutathione, for example, which is an endogenous antioxidant very important for red blood cells (and for other cell types...) and that reacts with peroxides undergoing oxidation. When it undergoes oxidation, it dimerizes with another oxidized glutathione. We also have some molecules that are exogenous antioxidants, namely vitamin C and vitamin E, which are very important antioxidants for our plasma and for our membranes, respectively. Note that there are many vitamins that do not have antioxidant function, that is, this characteristic can not be generalized to all other vitamins. There are also several antioxidants that are not indispensable to our metabolism, but they contribute to its good functioning, belonging to the class of bioactive compounds of the diet. Flavonoids or lycopene from tomatoes are good examples of this.
Therefore, if we look at the two classifications, it is easy to see that the exogenous antioxidants are always non-enzymatic, and that the endogenous antioxidants can be enzymatic or non-enzymatic. Regardless of the class where they are inserted, they are extremely important molecules and if we can guarantee an adequate contribution of them, surely we will be better prepared to deal with oxidative stress.

Metabolic map about the effects of nitric oxide (NO) on smooth muscle cells

Glucagon

Glucagon is derived from the Greek words gluco (glucose) and agon (agonist). It is a single-chain polypeptide with 29 amino acids, produced in the α-cells of the islets of Langerhans, located in the endocrine portion of the pancreas. This protein is important in the metabolism of carbohydrates. Its function is to increase glycemia by acting as an insulin antagonist. In a hypoglycaemia, glucagon is released into the bloodstream and acts mainly in the liver, where it binds to specific receptors on hepatocytes (which store glycogen), stimulating them to produce and then release glucose. This mechanism is called glycogenolysis. After glycogen stores cease, the liver synthesizes glucose through gluconeogenesis.Thus, under normal conditions, glucose ingestion inhibits glucagon secretion. During fasting, there is a decrease in hepatic glycogen, a decrease in glycolysis in the liver, a stimulation of gluconeogenesis, a stimulation of fatty acid oxidation in adipocytes and increase of serum levels of this protein. An important function of glucagon is to maintain the concentration of glucose high enough for the normal functioning of neurons, preventing seizures or hyporglycemic coma in normal fasting situations, such as in nighttime sleep.
Glucagon secretion is controlled physiologically not only by the hypoglycemia, but also by low levels of fatty acids, hyperaminoacidemia, vagal stimulation and adrenal system stimuli, such as stress or physical exercise. Increased glucagon in the blood will activate lipase from fat cells, inhibit the storage of triglycerides in the liver, inhibit the reabsorption of sodium by the kidneys, increase cardiac output, increase the secretion of bile and inhibit the secretion of gastric acid.
In the cases of pathology, high levels of glucagon in the blood may be present related to glucagonoma, a rare neoplasm of the α-cells of the pancreas, causing increased glucose and lipid levels, decreased levels of amino acids, anemia, diarrhea and weight loss. It is also observed the appearance of migratory erythema, characterized by the presence of erythematous blisters in the lower abdomen, buttocks, perineum and groin. Diabetes mellitus often results from the imbalance between the hormones insulin and glucagon present in this neoplasm.
Glucagon can be used in dental emergencies as in severe hypoglycemia, common in an uncontrolled diabetic. It can be administered intramuscularly, causing the rapid increase of glucose levels in the
blood.

Text written by:

- Catarina Capelo

- Dina Nair

- Marta Santos

- Samyra Matni

Cartoon about free radicals (2)

Oxidative stress (general considerations)

Today I decided to make a post on a very important topic, oxidative stress. This subject is often referred to in biochemistry classes (and not only!), but it is not always clear to the speaker or to the audience, what it actually represents.
Nevertheless, the idea is simple to understand ... As I say many times in my classes, we have a bad habit, which kills us slowly, without exception: we spend our lives breathing oxygen! And this molecule, so important for our biochemistry, in particular for aerobic metabolism, is what kills us slowly, and makes us grow old. And do not hesitate, if we do not die of an accident, or of some illness, we will die because we have been breathing O2 during our life! :)
So, what does oxygen contain that makes it so dangerous? Basically nothing, that is, the molecule itself is harmless to our molecules/cells. The problem is in its susceptibility to suffer partial reductions, that is, to capture electrons. In fact, we are continually forming the so-called reactive oxygen species, which are essentially 3: the hydroxyl radical (free radical), the superoxide anion (free radical) and hydrogen peroxide. Of these 3, the first two are more aggressive because they are free radicals. Free radicals are molecules that have an unpaired electron (which is why they are represented with a black speckle, which is that unpaired electron).  

The electrons have a serious problem, they do not like to walk alone, so they will look for "companionship" in the first molecule that appears ahead, be it a lipid, a protein or a nucleic acid. That is, reactive oxygen species are highly reactive molecules, they are powerful oxidizing agents, which will react with our biomolecules, removing an electron and altering/destroying them. And the problem is that although the free radical ceases to be when it picks up an electron, the molecule with which it reacts becomes a free radical, giving rise to a destructive chain process.
To counteract this, our cells have several defenses, called antioxidants. Therefore, oxidative stress arises when we have an imbalance between the pro-oxidants (reactive oxygen species and reactions that produce them) and the antioxidants (processes that prevent the formation and/or action of the pro-oxidants), favoring the first, or disfavoring the seconds.