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The Effects Of Metformin And Diabinese On Female Sex Hormone Of Type 2 Diabetes Mellitus Patients
[UNIVERSITY OF ILORIN TEACHING HOSPITAL (UITH), ILORIN, KWARA STATE]
CHAPTER THREE -- [Total Page(s) 6]
Page 3 of 6
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3.7.2 REAGENT
Materials provided with the kit:
1.
Antibody-Coated Wells (1 plate, 96 wells) Microtiter wells coated with
1.0 µg/ml streptavidin was packaged in an aluminium.
2. Reference Standard Set (1ml/vial) Contains 0, 20, 100, 250, 500 1500,3000 in pg/ml with preservatives.
3.
Rabbit Anti-estrogen Reagent (6 ml) Contains rabbit anti-estradiol
biotinylated rabbit igG conjugate in buffer, yellow dye with
preservatives.
4. Estradiol enzyme reagent (6 ml) Contains estradiol horseradish peroxides (HRP)
5. Substrate reagent (20 ml) contains TMB and hydrogen peroxide stabilized in buffer.
6. Stop Solution (8 ml) contains a strong Sulphuric acid(H2SO4)
7. Wash solution concentrate (20 ml) contains a surfactant in buffered saline.
8. Product instruction.
Materials required but not provided:
• Precision pipettes: 25 µl, 50 µl, 100 µl, 200 µl, and 1.0 ml.
• Disposable pipette tips.
• Distilled and deionized water.
• Vortex mixer or equivalent.
• Absorbent paper or paper towel.
• Linear-linear graph paper.
3.7.3 REAGENT PREPARATION
1. Working enzyme reagent:
0.7millitre of estrogen enzyme reagent was added to the vial containing steroid conjugate buffer.
2. Wash Buffer:
Wash solution was diluted to 1000 millitre with distilled in a suitable storage container.
3. Working substrate solution:
Amber
vial labelled solution A was poured into clear vial labelled solution
B. The yellow cap is placed on the clear vial for easy identification.
It was mixed and labelled accordingly.
3.7.4 ESTROGEN ASSAY PROCEDURE
• The micro plates wells were formatted in duplicate for each serum reference, control and patient specimen to be assayed.
• 25μl of the appropriate serum reference and specimen was pipette into the assigned wells.
• 50µl of Estradiol Biotin Reagent was added to all wells
• The microplate was swirl gently for 20seconds.
• It was incubated for 30 minutes at room temperature.
• 50µl of Estradiol Enzyme Reagent was added to all wells
• The microplate was swirl for 30seconds
• It was incubated for 90minutes at room temperature.
• The content of the micro plate was discarded by decantation and the plate was tapped and blotted dry with absorbent paper.
• 350ul of wash buffer was added, decant (tap and blot). This was repeated three times.
• 100ul of the working substrate solution was added to all the wells.
• It was incubated at room temperature for 20 minutes.
• 50ul of stop solution was added to each well and mixed gently for 15 seconds.
• The absorbance of each well was read at 450nm in a micro plate reader.
CHAPTER THREE -- [Total Page(s) 6]
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ABSRACT - [ Total Page(s): 1 ]ABSTRACT is coming soon ... Continue reading---
APPENDIX A - [ Total Page(s): 1 ]APPENDIX IQUESTIONNAIRE TO ACCESS THE ANTHROPOLOGIC AND DEMOGRAPHIC INDICES OF PATIENTS WITH TYPE TWO DIABETES MELLITUS ON ANTIDIABETIC DRUGS (METFORMIN AND DIABINESE) ATTENDING UITH ILORIN.INTRODUCTION: I am a final year students of the Department of Medical Laboratory Science, School of Basic Medical Sciences, Kwara State University, Malete, Kwara State. This questionnaire is aimed at accessing the demographic indices of patients with type 2 diabetes mellitus on metformin and diabinese in Ilor ... Continue reading---
APPENDIX B - [ Total Page(s): 1 ]APPENDIX 11REAGENT COMPOSITION FOR ESTROGENEstrogen calibrates, Horseradish Peroxidase Conjugates, Estrogen biotinylated purified rabbit igG conjugates, Tetremethylbenzidine(TMB), wash buffer concentrate, assay buffer, Stop solution.REAGENT COMPOSITION FOR PROGESTERONEProgesterone calibrates, Horseradish Peroxidase Conjugates, Progetarone biotinylated purified rabbit igG conjugates, Tetremethylbenzidine (TMB), wash buffer concentrate, assay buffer, Stop solution. ... Continue reading---
CHAPTER ONE - [ Total Page(s): 2 ]CHAPTER ONEINTRODUCTION1.1 BACKGROUND OF STUDY Diabetes
mellitus is a group of metabolic disorders in which a person has high
plasma glucose, either because the body does not produce enough insulin,
or because cells do not respond to the insulin that is produced. The
high plasma glucose produces the classical symptoms of polyuria,
polydipsia and polyphagia (Rother, 2007). Type 2 diabetes mellitus,
formerly non-insulin dependent diabetes mellitus or adult onset
diabetes, is a ... Continue reading---
CHAPTER TWO - [ Total Page(s): 17 ]CHAPTER THREE3.0 MATERIALS AND METHODS3.1 STUDY AREA The study was conducted in the Metropolitan City of Ilorin, Kwara State. Ilorin, the capital city of Kwara State, Nigeria is located on latitude 8o 24’N and 8o 36’N and longitude 4o 10’E and 4o 36’E with an area of about 100Km2 (Kwara State Diary, 1997). 3.2 STUDY POPULATIONThe population of this study was centred on patients with type 2 diabetes mellitus on anti-diab ... Continue reading---
CHAPTER FOUR - [ Total Page(s): 3 ]Table 4.4: Comparison of biochemical parameters (estrogen, progesterone and fasting blood sugar) between diabetic patients on metformin only and diabetic patients on combined metformin with other drugs (glimepride).Table 4.5 depicts the Pearson correlation of duration of diabetes and BMI with biochemical parameters (estrogen, progesterone and fasting blood sugar) in diabetic patients using antidiabetic drugs (metformin and diabinese).Data indicated a significant correlation between biochemical p ... Continue reading---
CHAPTER FIVE - [ Total Page(s): 2 ]CHAPTER FIVE5.0 DISCUSSIONType 2 diabetes mellitus, formerly known as non-insulin dependent diabetes mellitus or adult onset diabetes, is a metabolic disorder that is characterized by hyperglycemia in the context of insulin resistance and relative insulin deficiency (Vinay et al., 2008). Sex differences and the role of gonadal hormones in modulating insulin sensitivity and glucose tolerance are of increasing interest and importance because of the increasing prevalence of type 2 diabetes me ... Continue reading---
REFRENCES - [ Total Page(s): 4 ]Murali, R. and
Saravanan, R. (2012). Antidiabetic effect of d-limonene, a monoterpene
in streptozotocin-induced diabetic rats. Biomedical Prevention and
Nutrition.2:269-275.Neerati, P., Devde, R., and Gangi, A.K. (2014).
Evaluation of the effect of curcumin capsules on glyburide therapy in
patients with type-2 diabetes mellitus. Phytotherapeutic. Research.
28:1796-1800.Nelson, L.R. and Bulun, S.E. (2011).Estrogen production
and action. Journal of America Academic and Dermatology. 4 ... Continue reading---
