Assessment Of Pancreatic Hormones In Diabetes Mellitus & Non-diabetes Mellitus Patient: A Case-Control Study done on the group of people with unknown status.

1. Assessment Of Pancreatic Hormones In Diabetes
Mellitus & Non-diabetes Mellitus Patient: A Case Control
Study
Presented By : Rahul
Course & Semester : M.Sc. MLS-Biochemistry – 4th Semester.

2. OBJECTIVES OF PRESENTATION
• The Pancreas
• Anatomy & Histology of Pancreas
• Hormones of Endocrine Pancreas
• Insulin
• Glucagon

3. OBJECTIVES OF
PRESENTATION• Disorders of Pancreatic Hormones.
• Diabetes Mellitus -Types
• Signs & Symptoms
• Causes
• Aim of Study or Objectives of Study
• Materials & Methods
• Results
• Summary & Conclusion

4. OBJECTIVES OF STUDY
1. Screening of Diabetic and Non-Diabetic patients - based on
their HbA1c or sugar profile.
2. Estimation of Insulin and Glucagon in diabetes mellitus and
Non-diabetes mellitus patients.
3. Statistical analysis of the data.

5. PANCREAS
• The Endocrine system is the collection of glands that produce
hormones that regulates metabolism, growth and development tissue
function, sexual function, reproduction, sleep, and mood, among other
things.
• The word pancreas comes from the Greek language (pan means “all”)
& (keas means, “flesh”).
• The pancreas plays vital role in human body.
• The pancreas is an organ of the digestive system and endocrine
system of vertebrates.
• In humans, it is located in the abdomen behind the stomach and
functions as a gland.
• The pancreas has both an endocrine and a digestive exocrine function.Contd.

6. • As an endocrine gland, its functions mostly to regulate blood sugar , hormones like
insulin , glucagon , somatostatin, and pancreatic polypeptide.
• As a part of the digestive system it functions as an exocrine
gland secreting pancreatic juice into the duodenum through the pancreatic duct.
• This juice contains bicarbonate, which neutralizes acid entering the duodenum from the
stomach; and digestive enzymes, which breakdown carbohydrates, proteins, and fats in food
entering the duodenum from the stomach.
• Inflammation of the pancreas is known as pancreatitis, with common causes
including chronic alcohol use and gallstones. Because of its role in the regulation of
blood sugar, the pancreas is also a key organ in diabetes mellitus.
• The function of the pancreas in diabetes has been known since at least 1889, with its
role in insulin production identified in 1921.
Contd

7. ANATOMY & HISTOLOGY OF PANCRAS
• The pancreas is a long, slender organ, most of
which is located posterior to the bottom half
of the stomach.
• Although it is primarily an exocrine gland,
secreting a variety of digestive enzymes, the
pancreas has an endocrine function.
• Its pancreatic islets—clusters of cells
formerly known as the islets of Langerhans—
secrete the hormones glucagon, insulin,
somatostatin, and pancreatic polypeptide
(PP).
• The pancreas contain a Head , body, tail.
Contd

8. Islets of Langerhans play a crucial role in carbohydrate metabolism
and so in a plasma glucose concentration. It involves:
• Islets of Langerhans play a crucial role in carbohydrate metabolism and so in a plasma
glucose concentration. It involves:
• Glycolysis – the anaerobic conversion of glucose to lactate. Occurs in the red blood cells,
blood cells, renal medulla and skeletal muscles.
• Glycogenesis – the synthesis of glycogen from glucose. Glucose is stored (in liver,
liver, muscle) in the form of glycogen and this serves to maintain a constant plasma
glucose concentration.
• Glycogenolysis – the breakdown of glycogen to glucose.
• Gluconeogenesis – the production of glucose from non-sugar molecules (amino acids,
acids, lactate, glycerol)
• Lipolysis – the breakdown of triacylglycerols into glycerol and free fatty acids.
• Lipogenesis – the synthesis of triacylglycerols.
Contd

9. THE PANCREATIC ISLETS EACH CONTAIN FOUR VARIETIES OF
CELLS
CELL TYPES IN PANCREATIC ISLETS OF LANGERHANS.
Cell types
Approximate Percentage of Islet Volume
Secretory ProductsDorsally Derived
(Anterior Head, Body,
Tail)
Ventrally Derived
(Posterior Portion of
Head)
A cell (α) 10% < 0.5%
Glucagon, proglucagon,
glucagon-like peptides
(GLP-1 and GLP-2)
B cell (β) 70–80% 15–20%
Insulin, C peptide,
proinsulin, amylin, γ-
aminobutyric acid (GABA)
D cell (δ) 3–5% < 1% Somatostatin
PP cell (F cell) < 2% 80–85% Pancreatic polypeptide

10. HORMONES OF THE ENDOCRINE PANCREAS
HORMONES OF THE PANCREAS
Associated hormones Chemical class Effect
Insulin (beta cells) Protein Reduces blood glucose levels
Glucagon (alpha cells) Protein Increases blood glucose levels
Somatostatin (delta cells) Protein
Inhibits insulin and glucagon
release
Pancreatic polypeptide (PP
cells)
Protein Role in appetite

11. DISORDERS OF PANCREATIC HORMONES
• Insulin and glucagon, the two key hormones that orchestrate
fuel storage and utilization, are produced by the islet cells in
the pancreas. Islet cells are distributed in clusters
throughout the exocrine pancreas.
• Together, they comprise the endocrine pancreas. Diabetes
mellitus, a heterogeneous disorder, is the most common
disease of the endocrine pancreas
DIABETES MELLITUS (DM),
• commonly known as diabetes, is a group of metabolic
disorders characterized by a high blood sugar level over a
prolonged period of time.
Contd

12. • Diabetes is due to either the pancreas not producing enough insulin, or the cells of
the body not responding properly to the insulin produced.
• There are three main types of diabetes mellitus:
1. Type 1 diabetes results from the pancreas's failure to produce enough insulin due
insulin due to loss of beta cells, this form was previously referred to as "insulin-
dependent diabetes mellitus" (IDDM) or "juvenile diabetes.
2. Type 2 diabetes begins with insulin resistance, a condition in which cells fail to
fail to respond to insulin properly. As the disease progresses, a lack of insulin may
also develop This form was previously referred to as "non-insulin-dependent
diabetes mellitus" (NIDDM) or "adult-onset diabetes". The most common cause is
a combination of excessive body weight and insufficient exercise.
3. Gestational diabetes is the third main form, and occurs when pregnant
women without a previous history of diabetes develop high blood sugar levels.
Contd

13. SIGN AND SYMPTOMS
• Diabetes symptoms vary depending on how much your blood sugar is elevated.
Some people, especially those with prediabetes or type 2 diabetes, may not
experience symptoms initially. In type 1 diabetes, symptoms tend to come on
quickly and be more severe.
• Some of the signs and symptoms of type 1 and type 2 diabetes are:
• Increased thirst
• Frequent urination
• Extreme hunger
• Unexplained weight loss
• Presence of ketones in the urine Fatigue
• Irritability
• Blurred vision
• Slow-healing sores

14. PATHOPHYSIOLOGY
• Insulin is the principal hormone that regulates the uptake of glucose from
the blood into most cells of the body, especially liver, adipose tissue and
muscle, except smooth muscle, in which insulin acts via the IGF-1.
Therefore, deficiency of insulin or the insensitivity of its receptors play a
central role in all forms of diabetes mellitus.
• The body obtains glucose from three main sources: the intestinal absorption
of food; the breakdown of glycogen (glycogenolysis), the storage form of
glucose found in the liver; and gluconeogenesis, the generation of glucose
from non-carbohydrate substrates in the body.
• Insulin plays a critical role in regulating glucose levels in the body.
• Insulin can inhibit the breakdown of glycogen or the process of
gluconeogenesis.
Contd.

15. HYPOGLYCAEMIA,
• Its also known as low blood sugar, is a fall in blood sugar to levels below
• This may result in a variety of symptoms including clumsiness, trouble
confusion, loss of consciousness, seizures or death. A feeling of hunger,
shakiness and weakness may also be present. Symptoms typically come on
quickly.
HYPERGLYCAEMIA
• Its a condition in which an excessive amount of glucose circulates in
plasma.
• This is generally a blood sugar level higher than
symptoms may not start to become noticeable until even higher values
13.9–16.7 mmol/l (~250–300 mg/dl). A subject with a consistent range
~5.6 and ~7 mmol/l (100–126 mg/dl) (American Diabetes
is considered slightly hyperglycaemic, and above 7 mmol/l (126 mg/dl) is
generally held to have diabetes. Contd

16. MATERIAL & METHODS
• For carrying out above assessment the following materials are required and
the project will be carried out by collecting sample of 50 individuals &
their reports data.
• Material for Blood Collection
• Syringe with Needle
• Collection vial especially EDTA, Fluoride-grey top, Yellow top and Green
• top for Insulin.
• Tourniquet
• Alcohol Swab or cotton swab
• 70% Alcohol for cleaning of ante cubital fossa.
• Bandage
• Needle Burner or Needle destroyer
Contd

17. • Material for Hormonal Screening and
Diabetes Screening
• Reagent or chemical for glucagon & Insulin
• Test Tubes
• Micropipettes or Auto Pipettes (10-1000ul)
• GOD-POD Kit
• Centrifuge
• Tissue Paper
• Timer
Contd

18. PROCESS OF BLOOD COLLECTION &
SEPRATION

19. SAMPLE PROCESSING / METHODS
METHODS USED FOR GLUCOSE
ESTIMATION
• GOD-POD Method
Reaction :
D-glucose + O2+ H2O
𝑮𝑶𝑫
𝒈𝒍𝒖𝒄𝒐𝒏𝒊𝒄 𝒂𝒄𝒊𝒅 + 𝑯2O + phenol +
4-aminoantipyrine
𝑷𝑶𝑫
quinonimine
(red Color complex) + H2O.
• Normal values: -
• Fasting blood glucose – 70-110 mg/dl
• Postprandial (pp)- 110-140 mf/dl
METHOD OF GLYCATED HAEMOGLOBIN
• Using kit of Glycated Haemoglobin an
its measurement by semi
autoanalyzer.
Expected range of HbA1c
Sugar – 90-150 5-0% to 7.0%
Sugar – 150-180 7.0% to 8.0%
Sugar – 180-360 9.0% to 14.0%.

20. METHODS FOR HORMONES
• Method For Insulin
– Chemiluminescent Micro particle immunoassay.
• Insulin Test Preparation
• Insulin blood test or Fasting insulin test as the name suggests requires
fasting of minimum 8 hours or more as advised by the doctor.
• Insulin normal range: averages between 2.6 - 24.9 mcIU/mL

21. METHOD FOR GLUCAGON ESTIMATION
• A glucagon blood test measures the amount of a hormone called
glucagon in your blood. Glucagon is produced by cells in the pancreas.
It helps control your blood sugar level by increasing blood sugar when
it is too low.
• Method: RIA (Radio Immuno Assay)
• Normal Value: Normal levels are 150 to 200 pg./mL; in patients with
glucagonomas, levels usually (>90%) are higher than 1000 pg./mL.
However, in some recent studies, up to 40% of patients had plasma
glucagon values of 500 to 1000 pg./mL.

22. RESULTS & DISCUSSION
Total No. of
Samples N
Minimu
m
Maxim
um Mean
Std.
Deviati
on
Statist
ic
Statisti
c
Statisti
c
Statis
tic
Statisti
c
Fasting Blood
Glucose (
90-110 mg/dl) 50 85 250
131.
66 42.499
HbA1C
(4.5 - 5.6%) 50 5 9 6.06 1.219
Insulin
(2.6 - 24.9
mcIU/Ml) 50 4 50
19.3
4 10.497
Glucagon
(50 to 100
pg/ml) 50 51 190
88.9
8 38.682
NON
DIABETIC
N
Minim
um
Maxim
um
Me
an
Std.
Deviati
on
Stati
stic
Statis
tic
Statist
ic
Sta
tist
ic
Statisti
c
Fasting
Blood
Glucose
( 90-110
mg/dl) 22 85 120 97 8.106
HbA1C
(4.5 -
5.6%) 22 5 5 5 0
Insulin
(2.6 - 24.9
mcIU/Ml) 22 6 22
13.
36 4.583

23. DIABETIC
N Minimum Maximum Mean Std. Deviation
Statistic Statistic Statistic Statistic Statistic
Fasting Blood Glucose ( 90-110
mg/dl) 28 110 250 158.89 38.456
HbA1C (4.5 - 5.6%) 28 6 9 6.89 1.031
Insulin (2.6 - 24.9
mcIU/Ml) 28 4 50 24.04 11.475
Glucagon (50 to 100 pg/ml) 28 55 190 108.5 42.19
SUMMARY & CONCLUSION
In order to ensure normal body function, the human body is dependent on a tight
control of its blood glucose levels.
This is accomplished by a highly sophisticated network of various hormones and
neuropeptides released mainly from the brain, pancreas, liver, intestine as well as
adipose and muscle tissue.
Within this network, the pancreas represents a key player by secreting the blood
sugar-lowering hormone insulin and its opponent glucagon.

24. SUMMARY & CONCLUSION
• However, disturbances in the interplay of the hormones and peptides involved
may lead to metabolic disorders such as type 2 diabetes mellitus (T2DM) whose
prevalence, comorbidities and medical costs take on a dramatic scale.
• Therefore, it is of utmost importance to uncover and understand the
mechanisms underlying the various interactions to improve existing anti-
diabetic therapies and drugs on the one hand and to develop new therapeutic
approaches on the other.
• This review summarizes the interplay of the pancreas with various other organs
and tissues that maintain glucose homeostasis. Furthermore, anti-diabetic
drugs and their impact on signalling pathways underlying the network will be
discussed.
• In order to ensure normal body function, the human body is dependent on a
tight control of its blood glucose levels. This is accomplished by a highly
sophisticated network of various hormones and neuropeptides released mainly
from the brain, pancreas, liver, intestine as well as adipose and muscle tissue.
Within this network, the pancreas represents a key player by secreting the
blood sugar-lowering hormone insulin and its opponent glucagon.

25. • The conclusion of the study suggested that the ratio of samples was suggested that
the pancreatic hormones plays vital role in glucose storage and utilization. In type-
2 diabetes, both insufficient insulin and excessive glucagon secretion contribute to
hyperglycaemia. We compared insulin, glucagon and somatostatin stores in
pancreas obtained from different categories of patient such as obese non-diabetic
(ND), and 18 type-2 diabetic (T2D) subjects. From concentrations and pancreas
weight, total content of hormones was calculated. Insulin content was 35% lower in
T2D than ND subjects (7.4 versus 11.3 mg), whereas glucagon content was similar
(0.76 versus 0.81 mg). The higher ratio of glucagon/insulin contents in T2D was
thus explained by the decrease in insulin.
• To normalize metabolic control of glucose in the treatment of T2D, support has
increased for targeting not only abnormalities in insulin secretion but also
dysfunctional glucagon secretion. Glucagon is a key regulator of normal fuel
metabolism, and both fasting and postprandial hyperglucagonemia make
substantial contributions to the fasting hyperglycaemia and postprandial glucose
excursions that characterize T2D. Because patients with T2D have defects in
glucagon control, improved restoration of metabolic control by therapies that also
suppress glucagon, including DPP-4is and GLP-1RAs, would be beneficial. Future
studies should focus on how novel strategies such as glucagon antagonism,
glucagon/GLP-1 receptor coagonism, or combining DPP-4is or GLP-1RAs with
SGLT2is can best control both insulin and glucagon in patients with T2D.