By V. Delazar. George Washington University.
Because they are intrinsic constants order serpina 60caps amex anxiety level test, Ki values can theoretically be reproduced from one laboratory to another order serpina 60caps mastercard anxiety fatigue. The method of predicting the potential for drug interactions by a drug from Ki values and some measure of the in vivo concentrations of the drug is widely accepted (e purchase serpina 60 caps anxiety symptoms grief. Time-dependent inhibition occurs when the inhibitory potential of a drug can- didate increases as the enzyme is exposed to the inhibitor over time generic serpina 60 caps overnight delivery anxiety symptoms like heart attack. This type of inhibition may occur by several potential mechanisms, including the following: 1. Metabolism-dependent conversion of the drug candidate to a product that is a more potent direct-acting inhibitor than the parent (e. Metabolism-dependent conversion of the drug candidate to a metabolite that quasi-irreversibly coordinates with the heme iron (e. Slow-binding inhibition is a reversible process in which initial inhibition becomes more potent over time without any metabolism. Nonenzymatic degradation to inhibitory or reactive products can occur with some unstable compounds, such as rabeprazole, or some acyl glucuronides, which can rapidly rearrange to form reactive aldehydes that form Schiff’s bases (covalent In Vitro Study of Drug-Metabolizing Enzymes 253 adducts) with lysine residues on proteins (43). Inhibition that is only time dependent, such as slow-binding inhibition and the nonenzymatic formation of inhibitory products, are encountered less frequently than metabolism-dependent inhibition and will not be covered in detail in this chapter. However, by definition, the phrase “mechanism-based inhibition” excludes the formation of metabolites that are simply more potent direct-acting inhibitors than the parent, whereas the term “metabolism-dependent inhibition” includes this type of time-dependent inhibition. Strictly speaking, irreversible inhibitors that are affinity labeling agents, transition state analogs, and slow, tight-binding inhibitors (discussed above) are not mechanism-based inhibitors because they do not require a metabolic event to exert their effect (44). Any metabolite that is released from the active site cannot be the metab- olite that inactivates the enzyme. In such a case, inactivation may occur by binding to a site other than the active site, or by rearrangement of the metabolite prior to its return to the active site. Note that, in general, only a portion of the inactivator is converted to the species that covalently binds to the apoprotein or heme, and the rest is released from the active site. The amount converted to the inactivating species relative to other metabolites is known as the partition ratio and is generally >10 (i. The rate of inactivation is proportional to the concentration of the inactivator until all enzyme molecules are saturated, in accordance with Michaelis-Menten kinetics. Additionally, the decrease in enzymatic activity over time should follow pseudo-first-order kinetics. The addi- tion of an alternative substrate or competitive inhibitor with good affinity for the enzyme will prevent or at least decrease the rate of inactivation. There should be stoichiometric (ideally one-to-one) binding of inactivator to enzyme. Enzyme inactivation should be preceded by a catalytic event that converts the mechanism-based inhibitor to the inactivating metabolite. For most cases encountered in drug development, it is not necessary to definitively prove all of the above criteria. If necessary, subsequent experiments can then determine irreversibility and potency (e. This general experimental strategy, including follow-up experiments, is illustrated in Figure 5. The former tend to develop relatively quickly (in days or weeks) and, after the drug is discontinued, patients respond robustly when rechallenged with the same or a closely related drug, whereas the latter tend to develop rela- tively slowly (with symptoms sometimes appearing after six months or more of drug treatment) and patients may or may not respond to rechallenge with the drug. Uetrecht has proposed the general rule that drugs administered at a daily dose of 10 mg or less do not cause idiosyncratic toxicity (51). In 1995, alkylbenzene and 1-hexyne were reported to N-alkylate, the heme moiety of chloroperoxidase in a P450-like reaction (52). These compounds inactivated this heme-containing enzyme in a manner that met the criteria for a mechanism-based inhibitor as defined by Silverman (44). However, the inactivated enzyme spontaneously lost In Vitro Study of Drug-Metabolizing Enzymes 259 the heme adducts over several hours with a return of enzymatic activity and native heme, thus qualifying as “reversible inactivation. The heme adducts decomposed (became dealkylated) over time except under acidic con- ditions (53). However, Silverman noted that no period of time is defined for how long an enzyme-inactivator complex must persist for the inactivator to be classified as irreversible (44). In such cases, the inter- actions can persist for some time after cessation of the perpetrator drug. The kinetics of metabolism-dependent irreversible or quasi-irreversible inhibition can be complex (46) and are covered in chapter 11 of this book. The kinetics of metabolism-dependent inhibition caused by metabolites that function as direct-acting inhibitors (e. Therefore, when further examination of such inhibition is warranted, the inhibitory metabolite itself should be investigated. All of these designs offer certain advantages, but all have certain drawbacks as well, and most are intended to be used during drug discovery or early drug development. These may offer some advantages over fluorogenic substrates, but they have not been widely utilized (66,67) and appear more suited for screening during early discovery. Most of these substrates are not amenable to rapid analytical methods that make use of plate readers. Because of this limitation of chromatographic-based separations, several groups have developed radiometric assays based on radiolabeled conventional substrates, often with an extraction step, followed by scintillation counting (26,68–70). Test article interference is seldom a problem with radiometric methods, but their sensitivity varies with the specific activity of the substrate, which may necessitate the use of undesirably high protein concentrations and/or lengthy incubation times. In addition, radiometric methods are undesirable from a waste management per- spective. Two types of cocktail approach are used: one involves preincubation pooling of multiple marker substrates; the other involves postincubation pooling of multiple samples that were incubated with individual marker substrates. These problems are solved by incubating the marker substrates individually and pooling the samples after the incubations are terminated. However, a disadvantage of such postincubation pooling is that the samples become significantly diluted depending on the number of samples pooled. This controversy is partly because the principles of Michaelis- Menten enzyme kinetics (pure thoughts) are often applied to these (impure) systems. The advantages and disadvantages of each system are highlighted in the following sections. Another advantage is that the same sample of pooled human liver microsomes (and often the same experimental conditions, i. This is an important consideration because the enzyme that converts a drug to an inhibitory metabolite may not be the one that is inhibited (covered in detail below). A potential disadvantage of using pooled human liver microsomes is that these microsomes contain a large amount of lipid and protein that can decrease the free concentration of drug in the medium. However, to various degrees, this is a disad- vantage of all available in vitro systems. This disadvantage can be largely overcome by using highly sensitive analytical methods (e. Another potential disad- vantage is that human liver microsomes are an exhaustible resource; therefore, each batch of microsomes is slightly different, although the variability can be minimized by pooling samples from a large number of individuals and by preparing large batches with careful selection of individual samples. Indeed, when these measures are taken, pooled human liver microsomes may be one of the most consistent in vitro systems, with a well-designed pool lasting for four years or more (sufficient for 200 definitive studies, one per week or more). While the differences are sometimes artifacts of incubation conditions (especially those that are likely to violate the assumptions of the Michaelis-Menten equation), some differences appear to reflect genuine differences in the kinetics of reactions catalyzed by recombinant enzymes, purified enzymes, and human liver microsomes (Table 4).
Beta adrenergic receptor antagonists Physics of Circulation - Michael McConnell generic 60 caps serpina anxiety symptoms after eating, M buy discount serpina 60caps anxiety natural supplements. Non-linear aspects of the circulation best serpina 60 caps anxiety symptoms muscle twitching, including turbulent flow 60caps serpina overnight delivery anxiety symptoms 37, blood viscosity, and trans-capillary flow. The goal of these lectures is to apply several basic laws of physics toward understanding how blood flows and how the properties of the circulation - pressure, flow, velocity, vessel size, resistance, etc. The first step in understanding the physics of the circulation is to apply our intuition about the physics of solids to the physics of a fluid, in this case blood. Normally when we think of force, velocity, or potential and kinetic energy, we think in terms of a discrete, solid object. A fluid is more dynamic - it can change its shape and different parts can move at different velocities. Yet the same basic laws apply, it is simply a matter of expanding our intuition and terminology to describe the physics of fluid flow. While an object has a mass (m) and coefficient of friction (kf), a fluid will be described by its density (p) and viscosity g It is important to see that the laws have not changed, just the environment in which we apply them. As fluids are not discrete objects, it is more convenient to consider the force (F) per surface area (A), which is pressure (P). Consider the situation (fortunate or unfortunate) of 86 medical students piled into a sizable hot tub (like water in a beaker), as shown in the figure below. You would not have much difficulty reporting that there is a force acting on you due to the gravitational force on the mass above you. If we represent this mass by its density (p) and volume (V), we can say: F = mg = (V) g (m = V) Normalizing for the surface area at depth (d), which will be the cross-sectional area of the tub (A), we can derive the pressure (P): 3. While we use the geometry of the tub in the above example to illustrate the relationship of pressure to force, the pressure is actually independent of the geometry of the container. The figure below shows a variety of different geometries, and yet the height of each column is the same and the pressure at depth d is also identical. Pressure depends solely on the distance from the surface (d), as can be seen from the formula. The independence from specific geometry makes pressure an ideal variable to describe force on fluids. Just as force can cause the motion of an object, pressure can generate the flow of a fluid. Flow (Q) is defined as the amount, or volume (V), of fluid passing a point per unit time (t): Physics of Circulation - Michael McConnell, M. Velocity (v), on the other hand, is the speed with which individual elements of the fluid pass that point: v = x/t (units - m/sec) 3. Flow (Q) through a cylindrical tube is equal to the average velocity (v) of the fluid multiplied by the cross-sectional area (A) of the tube: Q = Av 4. For a closed-loop system, like the circulation, fluid cannot enter nor leave, so there is conservation of mass or a conservation of volume (assuming density is constant). Thus, the amount of blood that leaves the heart through the aorta must equal the amount of blood that flows through the capillary bed, which must equal the amount of blood that returns to the heart via the venae cavae (ignoring coronary and bronchial circulation). Otherwise there would be a build- up or loss of blood somewhere in the circulation. For example, the aorta has a high velocity and small area, whereas the capillary bed has a low velocity and large area. An important characteristic of the circulation is the resistance to blood flow through the vasculature. For an object sliding across a table, it is clear that the more friction the greater the force necessary to insure that the object makes it all the way across. Similarly, the more resistance to flow through the vasculature, the greater the pressure needed to drive blood from one end to the other. Thus, pressure (P) is directly related to resistance (R), in order to maintain a constant flow (Q): P α R (Q constant) 2. The circulation can actually be thought of as a circuit, with blood flow (Q) equivalent to electron flow or current (1), and pressure (P) equivalent to voltage (V). Notice that resistance (R) is used in both formulas - the resistance to blood flow through a vessel is analogous to the resistance to electron flow through an electrical resistor. Thus the vascular tree can be thought of as a large number of resistors, some in parallel and some in series. An important contrast between resistors in series and in parallel is that adding resistors in series always makes the total resistance larger whereas adding resistors in parallel always makes the total resistance smaller. The figures below show how the blood vessels and vascular beds are organized both in series and in parallel. Resistance is dependent on, and can be calculated from the physical properties of the fluid (i. In reality, fluid near the vessel wall is slowed dramatically as it has to move along a stationary surface (in the limit, the fluid at the wall has zero velocity). This slow moving outer layer of fluid then slows down the next layer closer to the center, setting up a "laminar" (i. The resulting velocity profile is parabolic or quadratic, as shown above,which means that velocity increases as a function of x2, where x is the distance from the vessel wall. Integrating the velocity profile allows us to derive the average velocity of the fluid. For this laminar flow, the average velocity (vave) is equal to half the velocity at the center (vmax), where x=r, the vessel radius. The overall result is that the average velocity (vave) is proportional to the vessel radius (r) squared. The other factors that affect velocity are the pressure (P) and viscosity g of the fluid, and the length (L) of the vessel. Pressure, as we have shown, is directly related to flow, which is directly related to velocity. The more viscous the fluid, the slower the velocity, so they are inversely related: v α 1/ 4. Finally, for a given pressure, the longer the vessel length (L) the less the velocity generated (think of how fast you can blow water out of a straw compared to a garden hose). For a cylindrical tube is, the exact relationship needs a factor of 1/8: vave = Pr2/8L 7. Note the fourth order dependence of flow (Q) on radius (r) - halving the radius reduces flow by a factor of 16! See if you can follow the calculation below, using the table below, to derive the relative resistance of the arterioles vs. Total Length Total (Mm) Cross- (Cm) Volume section (Cm²) al Area (Cm²) Aorta 10 1 0. To compare the resistance of the arteriolar bed to that of the capillary bed, we must start with the resistances of a single arteriole and capillary: Rart = (8/)Lart/rart4 Rcap = (8/)Lcap/rcap4 Taking the ratio between the two allows several factors to cancel out (we will assume that viscosity remains constant - a topic to be discussed later. However, the total resistance of a vascular bed is the combination of the resistances of all the individual vessels, which are organized in parallel. Thus, the total resistance is equal to the resistance of a single vessel divided by the number of vessels. Rart(total) = Rart/#arts Rcap(total) = Rcap/#caps Rart(total)/Rcap(total) = (Rart/Rcap)(#caps/#arts) Rart(total)/Rcap(total) = ( / )( / ) ~ 1. Given the larger cross-sectional area of the capillary bed and thus the much greater number of capillary vessels, the total resistance works out to be greater in the arteriolar bed by approximately 50%. Table 9 - Relative Resistance to Flow in the Vascular Bed: Calculated from Table 6 Poiseuille’s Law Aorta 4% Venules 4% Large arteries 5% Terminal veins 0. In the previous section we presented the conservation of mass and the resulting tradeoff between cross-sectional area and fluid velocity. In this section we will discuss the conservation of energy, where there is a balance between the potential energy and the kinetic energy of a fluid. This balance produces a tradeoff between pressure, a measure of potential energy, and velocity, a measure of kinetic energy.
They are well motivated to pre- vent the need for giving themselves daily shots of insulin cheap serpina 60 caps mastercard anxiety symptoms definition. Fried potatoes with 2 eggs (use only butter buy serpina 60caps on line anxiety symptoms explained, olive oil or lard) safe serpina 60caps anxiety upon waking, 1 cup hot or cold milk generic serpina 60caps mastercard anxiety attack symptoms. Cream of rice, with homemade “half n half” or whipping cream, cinnamon and vitamin C stirred in. Fruit cup, large bowl of peeled, chopped mixed fruit with whipping cream and 1 tbs. Green beans with potatoes, meat dish, cabbage apple salad, water with lemon juice and honey, 1 cup hot milk. Fresh green beans, especially fava beans contain a sub- stance that is described in old herbal literature to be espe- cially beneficial to diabetics. Potatoes (not overcooked), peeled to make sure there are no blemishes (contain mold and pesticide) can be cooked with the beans. Add fresh chopped parsley to the sauce or butter for both green beans and potatoes. Fresh parsley has special herbal goodness (high magnesium, high potassium, diuretic. Canned meat is safe from parasites but may have smoke flavoring added (contains benzopy- rene) or nitrates. Purchase the flip-top cans to avoid eating metal grindings from the can opening process. Add finely chopped apples (peeled) and a few apple seeds and whipping cream for the dressing. The drinking water should always have a little vitamin C, lemon juice or vinegar added, and 1 tsp. Asparagus, potato, raw salad, fowl dish, fruit, water with vinegar and honey, 1 cup hot milk. Fresh chopped chives may be added but no regular sour cream since this is very high in tyramine, a brain toxin. For dessert, fresh fruit chunks dipped in a homemade honey sauce (honey, water and cinnamon). The fruit may be chopped with whipping cream, cinna- mon and honey sauce (not more than 1 tbs. Acid foods stimulate; spices and B-vitamins (especially B ) stimulate; hot foods1 stimulate. Toxins at either location (especially food-derived toxins) tell the body to stop eating. Asparagus, meat dish, white rice (brown rice contains mold), coleslaw, milk, water, ice cream. A hot meat dish (no pasta, no wheat flour, no regular gravy) can be fried, cooked or baked, but not grilled. If more bread is requested, provide a wheat-free, corn-free variety; but limit bread eating to “after main dish” eating. If not enough milk is drunk: make custard pudding or rice pudding so the daily amount (3 cups) is consumed. There is no fruit or vegetable juice except homemade, and not much of that because it crowds out milk and water. If by chance, your elderly person hates these and starves themselves to get your sympathy, add a lot more potatoes and rice (never brown) to raise calories. The heavy use of cream and butter is offset by no deep fat fried food and little cheese. The morning blood sugar test is essential to keep track of changing circumstances. Be careful not to use rubbing alcohol when making the finger stick (use vodka or grain alcohol). Or even just the knowledge they are staying well controlled and will never have to take insulin shots. Diabetic Supplements Several supplements are especially good for diabetics: • Fenugreek seeds, 3 capsules with each meal. Maybe they have something in them that helps detoxify wood alcohol, since bilberry leaves are good for eyes, too. Diabetic Eating Out Since the rules are always somewhat relaxed when “eating out” a diabetic loved one will badger you to go out with them. If rules are sure to be broken, calculate it into the rest of the day so you can compensate for it. Ethnic foods often had to be given up when children were raised (switched to hot dogs and pizza) but with this diversion gone, a return to family food would be most welcome and most healthful. And they certainly were made at home where cleanliness and “persnickitiness” are at their finest! Good advice is to return to old fashioned home cooking: with its flour and butter, lard and cream, homemade pasta, olive oil and soup, coarse cereal grains and plain fruit. Gone are the fruit juices, flour mixes, crackers and sweets that fill grocery shelves. Time is the great inhibitor but if you have the means or the help, the best advice, nutritionally, is a return to old-fashioned cooking and recipes. Use her wooden spoons, glass glasses, and plain dishes, her wooden and straw bowls and enamel pots and pans. But a good salt rule is to either cook with it or have it on the table, but not both. Use aluminum- free sea salt, and make sure the salt is sterilized by heating five minutes at 400°F in a glass pie plate to kill mold. The best salt is a mixture of 1 part of your aluminum-free, sterilized sea salt and 1 part potassium chloride (another kind of salt, see Sources). Potassium ousts sodium (salt) from your body, so you can use twice as much of this kind of salt! It is important to find the poison as soon as you can since the rest of the body will soon be affected, too. It is a herculean task but only gets harder each day, so keep notes as you ask: Is there new carpeting? The list is endless and the situation looks hopeless because so many new things can happen in two weeks. To answer each question, test the item using your Syn- crometer searching technique. To test the air, take a dust sample off the kitchen counter or table (this gives you fresh dust). Be sure to test everything eaten in a two week time period: un- usual things like popcorn, candy, crackers, cookies, health foods and special powders. A consolation is that you will find a num- ber of bad foods that are not necessarily the tremor causes but which cause other health problems. Let us imagine that the air (dust) sample proves toxic (resonates with the saliva sample). Suppose the water proves toxic (appears in your white blood cells); search for lead, copper, and cadmium. Although municipal water tests occasionally detect small amounts of pro- pyl alcohol, benzene, or wood alcohol, I have never detected them—you need not search for them. Bacteria, coming from teeth and jaw (bone infections, called cavitations) may not seem as recent as two weeks.