DETERMINATION OF BLOOD SUGAR LEVEL IN A SELECTED SAMPLES OF URINE USING BENEDICT'S REAGENT SOLUTION

ABSTRACT
Introduction: True prevalence rate of diabetes mellitus in a population can be obtained by using invasive tests but it is practically difficult on large scale.
Aim: To find out the feasibility of mass non-invasive screening test to detect the prevalence of diabetes mellitus in rural population of India with the help of a mathematical formula.
Materials and Methods: From population of 18800 residing in two adjacent rural areas of Uyo, a systematic random sample of 1005 adult subjects was screened for diabetes by using urine benedicts test, Canrisk questionnaire, Madras Diabetes Research Foundation-Indian Diabetic Risk Score (MDRF-IDRS) and determined prevalence of diabetes (pA) gauzed by each of these screening tests. Simultaneously, each subject’s glycaemic status was confirmed by standard fasting Plasma glucose (FPG) and postprandial plasma glucose (PPPG) levels. The blood test was also used to determine true prevalence which was crosschecked with the prevalence estimated (Pe) by the above stated screening tests using a mathematical formula.
results:  The true prevalence of T2DM in more than 18 years of population by Fasting Plasma Sugar (FPS) was 4.5% while that by using mathematical formulae that estimated by urine test, Canrisk test and MDRF-IDRS was 4.4%, 4.4 and 4.3% respectively.  When more than 35 years age-group was selected, true prevalence was 7.4% and estimated prevalence by Canrisk test was 7.1% (as against gold standard of Fasting) and 6.9% (as against PP). By fasting urine test it came out to be 7.2% and by PP urine test it was 7.4%. In population l8-35 years, the prevalence of diabetes was 1.1% by plasma glucose test. By using Canrisk, it came out to be 1.04%.

Keywords: Canrisk, Uyo, Mathematical modeling, Non-invasive, Rural population



CHAPTER ONE
INTRODUCTION
Background to study
Diabetes is the most common chronic disease among metabolism and endocrine diseases. Diabetes is becoming a major medical problem, causing a number of socially detrimental effects such as increasing the burden of medical costs, reducing social labour, increasing the rate of death and shorten the life of the patient.
 Nowadays, diabetes is increasing in developed countries, where urbanization gradually changes the lifestyle, eating habits and physical activity. Diabetes mellitus is associated with many chronic and acute complications of cardiovascular complications. These complications, together with psychological stress, not only diminish the quality of life of the patient but also endangers life expectancy, leaving many serious and permanent sequelae, resulting in increased mortality. For that reason, it can be seen that the high diabetes rate is a burden to the community and society.
 It was found that from 40 to 49-year-old with a diagnosis of type 2 diabetes would lose an average of ten years of life. Patients with diabetes mellitus have 2-3 times the risk of coronary heart disease than those without diabetes. On the other hand, at the time of diagnosis, the majority of diabetic patients had complications, including retinopathy of 35%, peripheral neuropathy 12%. According to the World Health Organization (WHO), in 2008 the world had 135 million diabetics (4%) of the world population, only two years (2010) the number of people with diabetes to 221 million (5.4%). In Vietnam, in recent years, the rapid development of diabetes has become a major problem in the health sector. According to calculations by the Association of diabetes educators in Vietnam, the rate of diabetes in 2002 accounted for 2.7% of the population, by 2008 (after 6 years) has doubled 5.7% of the population. (Timon, Collantes, Galindo & Gomez, 2014.)
 Blood sugar or blood glucose is a basic test used to assess glucose metabolism disorders such as diabetes mellitus or hypoglycaemia. Testing is done by two methods which are chemistry and enzyme. However, chemical methods are currently unavailable for measuring non-specific and high time-consuming glucose levels. Instead, nowadays the most common method is to use enzymes. The Benedict’s reagent solution has higher specificity and time is also faster, giving the best results. Currently, the three most commonly method used enzyme to quantify blood glucose that are hexokinase, glucose oxidase and glucose dehydrogenase. This research focus primarily on the use of glucose oxidase enzymes to quantify glucose in the blood. This is a common method with chemistries for semi-automatic biochemistry and some biochemical machines. This method combines the use of glucose oxidase and peroxidase.
In the body, glucose is known as an important carbohydrate source for energy. The glucose concentrations are maintained by gluconeogenesis and glycogenolysis in starving state while the glucose circulating is found in the fed state. The majority amount of glucose is found in complex carbohydrates which are separated to monosaccharides during the digestive period while a small amount of glucose is performed in fed state as glucose. (McMillin, 1975.) This section will show the glucose transporter and the way to control the blood glucose. 


1.1 Glucose transporter
 Glucose is used effectively by a significant number of different cell types under normal conditions, however, its content in the blood must be controlled accurately. Glucose plays a central role in the metabolism and homeostasis of the cell. Most of the cells in the body need continuous supply of glucose in the form of ATP to provide energy. The glucose balance disorders are known as a reason of the diabetes. Glucose is absorbed to the cells through the cell membrane. Molybdenum molecules cannot travel through the cell membrane by diffusing simply because the high molecular pathway cannot pass through the infinity matrix of the double lipid phosphorus layer. An efficient transport system is required to move molecules into and out of the cell for the glucose molecules used by cells. In certain absorbed cells, such as epithelial cells of the small intestine and tubules, glucose crosses the cell membrane (active transport) against the concentration gradient, injected by Na+/K channel system. However, glucose is transported passively to almost all cells in the body by a mediated transport mechanism without energy. The transporter protein involved in this process is called glucose transport, abbreviated as GLUT. (Wood &Trayhurn, 2003.)
There are twelve different transport proteins have been identified along with their genetic codes. Genome project assisted in this identification because all transport proteins share a similar structure and sequences in their genomes. Approximately 28% of amino acids are common in the transporter protein group. Each

GLUT is a protein that separates, penetrates and extends the lipid bilayer of the cell membrane and pass through the membrane several times. (Goodsell, 2017.) The pattern of glucose transport in the molecular space is illustrated in Figure 1. 

In the simplest form, a transport protein has a specific location for the molecule being transported. Moreover, it also undergoes a configuration change when it binds to the molecule, allowing the molecule to pass through the other side of the membrane and be released. Protein transport is also capable of reversing process can be repeated. All cells express at least one GLUT isotope on their plasma membrane. Other isoforms have distinct tissue distribution and cell biochemistry, and they contribute to the precise processing of glucose under different physiological conditions. (Wood &Trayhurn, 2003.) TABLE 1: Twelve isoforms of glucose transport. (Dam, 2018.) 


GLUT 1 is expressed in the red blood cells and endothelial cells of the brain with the responsibility of providing basic glucose to the cells. GLUT 2 is involved in the transport of glucose from the intestinal mucosa into the bloodstream via the portal and, moreover, it can also transport fructose from the intestinal lining cells, the rate of transportation dependent on glucose concentration in the blood. GLUT 3 is considered a highly valued tissues dependent on glucose concentration such as the brain.  In contrast, GLUT4 is quite sensitive with insulin and concentrate on the cell membrane that increases the hormones. The increase glucose transport across the membrane is accompanied by an increase in glucose uptake by insulin stimulating cells. The presence of GLUT 4 in skeletal muscle and in fat tissue, makes these tissues respond to insulin. The liver, brain, and erythrocytes lack GLUT 4, and, therefore, insensitive to insulin. Besides, fructose transport is characteristic of GLUT 5. (Navale&Paranjape, 2016.)
Molecular biology techniques have been applied to the study of the activity of some GLUTs. GLUT has also been found at glucose level in certain tissues such as stem cells of the pancreas. Moreover, following ribosomal mRNA synthesis on the endoplasmic reticulum, the protein transported into the Golgi system, where it was synthesized in the tubular structure of the trans-Golgi network. In the absence of oocyte stimulation, GLUT4 is located in these structures as well as in cytoplasmic pouches. In skeletal muscle cells, there is also the distribution of GLUT 4. The balance between intestinal absorption of glucose and the absorption and metabolism of peripheral tissues have maintained the blood glucose level within a narrow range. (Navale&Paranjape, 2016.)
1.1 Insulin Insulin plays an important role in the metabolism of lipids and amino acids. It is a powerful anabolic hormone and involved in the synthesis, and storage of glucose, lipids and amino acids or protein. Moreover, insulin promotes the expression or activity of enzymes that catalyse the synthesis of glycogen, lipids, and protein. The enzymes that catalyse the metabolism of glycogen, lipids, and amino acids, are inhibits the expression or activity. (Ogbru& Williams, 2016.) The anabolic and catabolic effects of insulin on glucose and glycogen, fatty acids and triacylglycerols, and amino acids and proteins is illustrated in Figure 2.....

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Item Type: Project Material  |  Size: 35 pages  |  Chapters: 1-5
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