INTRODUCTION: Lactate dehydrogenase (LDH) is a tetrameric molecule categorised into several types, which are known as isozymes or isoenzymes. LDH activity is commonplace in all tissue and can be seen in higher quantities in the blood when tissue damage is present due to enzyme leakage, typically seen in myocardial infraction or liver disease (1, 2). LDH consists of either a heart (H) or a muscle (M) type subunit and can be made up collectively where five different proteins may be produced H4, M4, M2H2, M3H and MH3. Migration rates differ with H4 having the highest and M4 the lowest rate toward the anode (1). H is seen in highly oxidative tissues, where damage leads to leaky tissue, as seen in a myocardial infarction (MI), while tissue which …show more content…
Although they differ in their molecular structure, they catalyse the same biochemical reaction (4). Heart, liver, and muscle cells typically have an abundance of LDH, where we see a reversible, lactate and pyruvate catalytic reaction occurring (5). Part 1, experimental data (Figure 2) was unable to be used as no results were successfully attained. MD results (Figure 3) were used for the remainder of the experiment. Electrophoresis showed no bands in lanes 1 & 5 plasma, indicating the LDH levels were not high enough to be detected. Lanes 2 & 6 showed darker bands at the anode end, which is to be expected due to heart being a highly oxygenated tissue and having a high concentration of H4 isoenzymes. Liver tissue shown in lanes 3 & 7, indicates a lower level of NADH production seen by the faint bands, with the bands at the cathode end being more visible, this is due, to there being more M type being present. The bands at the cathode end seen in lanes 4 & 8 all show a dark band. This could be due to skeletal muscle being less oxygenated, therefore, having a more positive charge (2). Electrophoresis is a favourable technique to observe the five isoenzymes to see the production of NADH (1,
During a moderate exercise, the rate of lactate in arterial blood is superior to three
It should be noted that the focus of this paper will be on CHD due to atherosclerosis, which makes up over 90% of all ischaemic heart diseases [1].
Cellular hypoxemia is resultant of inadequate amounts of oxygen being delivered to the cells or the inability of the cells to use the oxygen. This can be caused by ischemia, respiratory disease, vasoconstriction, vascular obstruction, edema, or anemia. Hypoxemia may result in power failure in the cell resulting in cell death. As the oxygen tension in the cell increases, anaerobic metabolism begins building up lactic acid and reducing pH levels causing biochemical reactions, chromatin clumping, and cell shrinkage. The leakage of intracellular enzymes into extracellular fluid is an indicator of cell injury or death (Grossman & Mattson Porth, 2014). This is evident with the edema Maria is
The importance of impaired lactate metabolism is illustrated by the observation that, when lactic acid is infused into normal animals at rates similar to the rate of overproduction that occurs in shock, hepatic utilization of lactate increases, and there is only a minimal fall in arterial pH [5]. In shock, reduced hepatic perfusion and an associated intracellular acidosis probably combine to substantially diminish hepatic lactate metabolism [4,5,12].
Introduction In order to digest the lactose that milk has the enzyme lactase is needed to break it down. Lactase is a large glycoprotein and has two active sites. These active sites can activate and determine a variety of β glucoside, β galactoside, lactose, and others (Swallow). The enzyme lactase is a dominant genetic trait that is inherited and is known as lactase persistence.
Lactic Acid produced in muscle cells is transported through the bloodstream. It is transported to the liver, where it is converted back into pyruvate.
Presence of high levels of these enzymes in the bloodstream is the standard test for damage to the heart.
This free radical, which is initially formed as relatively unreactive, reacts very rapidly with oxygen to yield a highly reactive trichloromethyl peroxy radical (CCl3OO•). Both radicals are capable of binding to proteins or lipids, or abstracting a hydrogen atom from an unsaturated lipid, thus, initiating lipid peroxidation (Halliwell, and Gutteridge, 1990; Williams, and Burk, 1990; Lee, and Jeong, 2002). Lipid peroxidation may cause peroxidative tissue damage in inflammation, cancer, aging, ulcer, cirrhosis, and atherosclerosis. Therefore, inhibition of the cytochrome P450-dependent oxygenase activity could cause a reduction in the level of toxic reactive metabolites and a decrease in tissue injury. On the other hand, an elevation of plasma AST and ALT activities could be regarded as a sign of damage to the liver cell
The main purpose of the lab was to investigate the effects of lactase two sugars, lactose and maltose. It was concluded that adding lactase to lactose will cause it to break down into its two components and this was indicated by adding benedict's solution. In the presence of glucose, benedict's solution causes the color to change to a red-yellow solution, which is what was observed. Gene expression can be used to
A continual production of lactic acid through fermentation may result in an increase of acidity within muscle cells and blood.
Principle Hexokinase is an enzyme that catalyzes the reaction between ATP and glucose. This reaction produces ADP and glucose-6-phosphate. When glucose -6 –phosphate is present with NAD it is oxidized by glucose-6-phosphate dehydrogenase to a phosphogluconate and NADH. The concentration of NADH is proportional to the glucose concentrations and can be measured by a spectrophotometer at 340nm (CDC,2003-2004).
Lactate dehydrogenase FUNCTION : Catalyzes enzyme that transfer a hydrogen from molecule to another molecule, which converts pyruvate to lactate and also converts NADH to NAD+ Location : found near all living cells and there is 4 different enzyme classes, 2 are cytochrome c-dependent enzyme (D-lactate or L-lactate) and 2 are NAD (P)- dependent enzyme (D-lactate or L-lactate) ` ` Reaction : Lactate dehydrogenase catalyzes the interconversion of pyruvate and lactate with associated interconversion of NADH and NAD+. It converts pyruvate to lactate in the ending product of glycolysis with absent of oxygen or in little supply, and it performs the reverse reaction during the Cori cycle in the liver. At high concentrations of lactate, the enzyme exhibits reaction inhibition, and decrease the
Three bands were distinguished (not including the standard curve bands), one band was present in the crude fraction lane and two bands in the mitochondrial fraction lane. No protein bands were present in the Nuclear fraction lane nor the soluble fraction lane. The Cox9 Ab binds to mitochondria so it was expected that the mitochondrial fraction lane would develop the most bands. Most of the mitochondria should be in the mitochondrial fraction. (Hart, 2017) The crude fraction lane also had mitochondria that interacted with Cox9 Ab, therefore, subcellular mitochondria was in the crude fraction, to a lesser extent. It was evaluated that the mitochondrial fraction proteins had greater migration (Table 9, 10, and 11) and density in comparison to the crude fraction lane. The ability to mark mitochondria with an Ab for further study is used frequently in scientific experimentation. (Edwards, Rawsthorne, & Mullineaux, 1990) Cell fractionation, SDS-PAGE, and Western blotting, is specifically used to study enzymatic activity. The mitochondria marked in the mitochondrial fraction lane could be collected for further purification and
Foster went on to explain the mitochondrial CPT system that occurs in the liver. The enzyme that has the most control on this system is called carnitine transferase 1. This is the enzyme that puts the carnitine on the long chain fatty acid and then the whole thing goes through the membrane. When inside the membrane of the mitochondria, CPT 2 puts the CoA back on. And the acyl (or acetyl) CoA is converted into a ketone body in the liver. In addition, the regulatory molecule of this system is Malonyl CoA. It has the ability to inhibit CPT1. In fed state malonyl co A is built right back up and the liver only makes L.DL from long chain fatty acids. However, Malonyl CoA is controlled primarily by two enzymes, osteocolay carboxylase which makes it and puts a Co2 on an acetate molecule, and broken down by Malonyl CoA decarboxylase. It has a dual function that is being the primary substrate from which make long chain fatty acids and at the same time inhibits fatty acid oxidation. Mal co A decarboxylase is an activatable enzyme. Functioned on by kinase, if phosphorylate it by glucagon it becomes inactive, things drop. Will give you fall of Malonyl CoA. But the synthesis side is the more important regulator in most tissues.
In order to evaluate the myocardial tissue alterations in the xenograft model, rats were killed at (24) post Qingyi Decoction treating and left ventricular myocardial tissues were collected from each group and fixed in 10% formalin, processed and embedded in paraffin wax. Thin sections of 3-5 microns thickness were cut and placed on microscopic slides. The tissues were deparaffinized in xylene solution, rehydrated in downstream serial dilutions of ethanol and stained with hematoxylin for 10 minutes, bluing for 10 minutes in running tap water, decolorized for 3 seconds by 1% acid alcohol, and the tissues were stained with eosin for 1 minute, washed with water and let to air dry. Fix the cover slips on the tissues. The slides were examined under a light microscope at a magnification of 40x, to detect the histological changes in the left ventricular myocardial tissues of each experimental groups