CHAPTER I

THE PROBLEM AND ITS SETTING

Introduction:
Diverse ailments are consistently increasing due to unconscious and unhealthy lifestyle of
people nowadays. Diseases have already been part of the everyday existence of peoples lives. It
has been construed that most diseases are acquired through heredity and environmental factors.
While population is perpetually increasing, the environment is also changing which becomes a
contributing factor of peoples sickness. This also includes the kind of food that people
consumes. Unwise and unhealthy consumption of food may put someones health at risk.
Diabetes is one of the common diseases that concern the society. It has been considered
as the leading diseases among individuals that if left untreated will lead to death.
However, as we go through modernization, Science, with the aid of the new technologies,
contributed in formulating different remedies for this particular ailment. New studies have been
initiated to examine on how to treat this kind disease and how it can be prevented at the early
stage. This is why modern researchers have been encouraged to use herbal medicines as an
alternative prevention for person with diabetes.
Noni (Morindacitrifolia)is a tree in the coffee family, Rubiaceae. Its native range extends
through Southeast Asia and Australia. Noniis among the medicinal plant that has been used in
medicinal research. Its fruit and leaves extract is used to treat wide range of maladies. Hence,
Noni (Morindacitrifolia) plant may live in tropical countries like the Philippines; it has been
sought out as an alternative for costly medicine.

This research investigates the efficient, viable, and affordable techniques in lowering
blood glucose levels. Aside from this, it will also assess the capability of Noni
(Morindacitrifolia) leaves extract as a viable remedy for diabetes that is caused by elevated
blood glucose.
Background of the Study:
High level of blood glucose may be known as the common problem people are facing.
Needless to say, people are consuming food with high sugar and fats. They are unaware of the
effects of the food they intake which has become the reason for acquiring this disease. Because
of this, researchers have been studying and formulating different substances to possibly prevent
the said disease with the use of medicinal plants.
The use of plants as remedial treatments has been commonly used among indigenous
people. Moreover, people living in remote areas are also adapting herbal medicines to cure
particular disease which is effective in many cases.
With this in mind, the researchers are motivated to explore whether the Noni
(Morindacitrifolia) leaves has a property or component that can lower the blood glucose level. In
order to test the plants effectiveness in lowering high blood glucose levels, Swiss Albino mice
(Musmusculus) are used as part of this experiment. This is so to determine the capability of Noni
(Morindacitrifolia) as a way to prevent high blood glucose.




Statement of the Problem:
This research aims to determine the effectiveness of Noni (Morindacitrifolia) leaves
extract as an agent in lowering blood glucose level if it is induced to Swiss Albino mice
(Musmusculus). Specifically, this research aims to answer the following questions:
1. The effect on the blood glucose level of Swiss Albino mice (Musmusculus) of the
Noni (Morindacitrifolia) leaves extract.
2. The precise level of concentration of Noni (Morindacitrifolia) leaves extract
inhibiting blood glucose level of Swiss Albino mice (Musmusculus).
Objectives:
This study aims to:
1. General Objective
a. Produce a cheap alternative remedy for diabetes.
b. Promote the significance and effectiveness of Noni (Morindacitrifolia) in our
community.
2. Specific Objective
a. Test the effectiveness of Noni (Morindacitrifolia) in lowering blood glucose.
b. The best concentration in lowering blood glucose level.
Hypotheses:
The researchers are guided by the hypotheses stated in null and alternative forms and is
tested at 0.05 level of significance.

H
o
= Noni (Morindacitrifolia) leaves extract cannot lower high level of glucose of Swiss
Albino mice (Musmusculus).
H
a
= Noni (Morindacitrifolia) leaves extract can lower high level of glucose of Swiss
Albino mice (Musmusculus).

Significance of the Study:
The importance of the study is to find new alternative medical solution using plants that
will contribute to vary the problems our society is facing these days.
The study as a whole is beneficial to the following:
The Society whose constituents are mostly affected by diabetes because of unhealthy
lifestyle.
The Government will no longer spend more funds to costly medicines and may
increase their revenues through this Noni product.
The Farmers may also encourage planting Noni tress as part of their financial
earnings.
The Students and Researchers may have indulged to study further the other beneficial
that a Noni fruit has.



Scope and Limitations:
The study determines how efficient the Noni (Morindacitrifolia) leaves extract is in
lowering the blood glucose level of Swiss Albino mice (Musmusculus). Though Noni
(Morindacitrifolia) is used to treat wide range of maladies, this study is limited only for diabetes
as well as the utilization of the leaves of the Noni (Morindacitrifolia). The Noni
(Morindacitrifolia) leaves extract were reaped near the Satellite, Pacita, San Pedro, Laguna.
The Swiss Albino mice (Musmusculus) were acquired at the pet shop near San Pedro
College of Business Administration (SPCBA). The gender of the mice were not considered, and
only two trials are used; the control set-up and the experimental groups which is tested during the
experimentation. Since, Swiss Albino mice (Musmusculus) has the same composition organ as
human, its use for this experiment is needed. Hence, this study is restricted to the glycaemic
response of Swiss Albino mice (Musmusculus) in Noni (Morindacitrifolia). It doesnt include the
relation of the Swiss Albino mice (Musmusculus) on other ailments.
Conceptual Paradigm:
Figure 1
Noni (Morindacitrifolia) Leaves Extract in Lowering Blood Glucose Level of Swiss Albino
mice (Musmusculus)


INPUT THRU-PUT OUTPUT
Extraction of Noni
(Morindacitrifolia)
leaves
Application of Noni
leaves extract to Swiss
Albino mice
Glycogenic response of
Swiss Albino mice to
Noni leaves extract

Figure 1 show the research paradigm used in the study. The input of the study includes
the extraction of Noni (Morindacitrifolia). The thru-put involves the application of Noni
(Morindacitrifolia) leaves extract to Swiss Albino mice (Musmusculus). The output involves the
glycogenic response of Swiss Albino mice (Musmusculus) to Noni (Morindacitrifolia) leaves
extract.

Definition of Terms:
Blood Glucose Level
Amount of glucose (sugar) present in blood
Chocolate
A raw or processed food produced from the seed of the tropical Theobroma
Cacao tree
Diabetes
Metabolic diseases in which the person has high blood glucose (blood sugar)
Glucose
A simple sugar in human metabolism
Hyperglycemia (also spelled as Hyperglycaemia)
A condition in which an excessive amount of glucose circulates in the blood
plasma or high blood sugar

Hyperinsulinemia
Raised blood levels of insulin
Hypoglycemia (also spelled as Hypoglycaemia)
An abnormally diminished content of glucose in the blood or low blood sugar

Medicinal plants
Alternative remedy for ailments, using plants, fruits, leaves, seeds, roots as
medicinal purposes
Noni (Morindacitrifolia)
A type of evergreen ranging in size from a small bush to a tree 20 or 30 feet high,
has a lumpy appearance with a waxy, semi-translucent skin that ranges in color.
Swiss Albino mice (Musmusculus)
Mice that has the same organ composition as human; commonly used in
experiments.





CHAPTER II
REVIEW OF RELATED LITERATURE AND STUDIES
This chapter presents the review of related literature and studies complied by the
researchers from different sources like books magazines and also the internet.
REVIEW OF RELATED LITERATURE
Foreign
Diabetes (diabetes mellitus) affects approximately 135 million people in the world.
Diabetes and hypertension are both diseases in westernized countries. Both entities increase with
age.
Diabetes (diabetes mellitus) is classed as a metabolism disorder. Metabolism refers to the
way our bodies use digested food for energy and growth. Most of what we eat is broken down
into glucose. Glucose is a form of sugar in the blood and it is principal source of fuel for our
bodies.
In ancient China, people observed that ants would be attracted to some peoples urine,
because it was sweet. The term Sweet Urine Disease was coined.
According to a press release issued by the National Institutes of Allergy and Infectious
Diseases and a report in this weeks edition of the New England Journal of Medicine (NEJM), a
transplantation technique known as the Edmonton Protocol may be of significant benefit to
patients with type I diabetes.

There are two types of diabetes mellitus, the type 1 diabetes, no insulin is produced,
meaning the glucose is not taken up from the blood, leading to a high blood sugar level
or hyperglycemia.Type I is an autoimmune disease (a condition that occurs when the bodys
immune system malfunction and attacks particular types of the bodys cell as if they were
foreign cells) in which the islet cells of the pancreas are destroyed by antibodies produced by
the bodys own faulty immune system. Type I diabetes is also known as Insulin-Dependent
diabetes because those with this condition require daily injections of insulin to survive.
In type 2 diabetes, blood sugar levels are also raised, because an inadequate amount of
insulin is produced to facilitate the cellular uptake of glucose by the body's cells. Alternatively,
insulin levels may be sufficient, but the body cells fail to respond normally to the presence of
insulin, even when levels are raised. This is called insulin resistance.Insulin resistance describes
when the body produces insulin but the usual actions of insulin are not mediated.
The resulting lack of glucose in cells triggers the pancreatic cells to produce
more insulinstill, resulting in raised blood levels of insulin or hyperinsulinemia. Therefore, type
1 diabetes is caused by an actual deficiency in insulin while type 2 is caused by a relative
deficiency.
Around 75% of people with diabetes have type 2 diabetes mellitus. This has also been
called non-insulin dependent diabetes mellitus (NIDDM) or maturity-onset diabetes mellitus.
The incidence of type 2 diabetes is on the increase worldwide as the number of health problems
such as obesity increase. In type 2 diabetes, not enough insulin is produced or the insulin that is
made by the body does not exert the usual effects on body cells.



Local
Noni (Morindacitrifolia), also known as bangkoro, is as type of evergreen ranging in size
from a small bush to a tree 20 or 30 feet high that grows in open coastal regions from sea level to
elevations of 1300 feet. The Noni fruit is approximately the size of a potato and has a lumpy
appearance with a waxy, semi-translucent skin that ranges in color as it ripens from green, to
yellow, to almost white when it is ripe and ready for juicing.
Hypoglycemia (also spelled as Hypoglycaemia) is an abnormally diminished content of
glucose in the blood or low blood sugar while Hyperglycemia (also spelled as
Hyperglycaemia) is a condition in which an excessive amount of glucose circulates in the blood
plasma or high blood sugar.
REVIEW OF RELATED STUDIES
Foreign
Based on the Epidemiologists from the School of Medicine at The University of Texas
Health Science Center San Antonio, diet soft drink consumption is associated with increased
waist circumference in humans, and a second study that found aspartame raised fasting glucose
(blood sugar) in diabetes-prone mice.
Elevated glucose levels are most strongly link to pancreatic cancer, doubling the risk of
death from the malignancy in both genders. Other cancers significantly linked to high glucose
levels induced esophagus, liver, and colon/rectum in men and liver and cervix in women.


Local
Noni (Morindacitrifolia), in the Philippines it is growing in coastal areas of Bohol,
Batangas and Bataan, probably present in a lot more of the coastal provinces. It is sadi that all
parts of Noni is medicinal but recently the fruit was tapped as a dietary supplement for general
well-being.
Based on agris.fao.org, Annie Evangelista from University of the Philippines, Los Banos,
Laguna (UPLB) studied about tissue culture as an alternative propagation technique of this plant.
Young leaves from shoots of rooted large diameter-stems were the source of explants. For callus
induction, sterilized explants were grown in agar-solidified Murashige&Skoog (MS) media with
2% sucrose and 1.0 and 10 microM each of 2,4-D in BAP [benzylaminopurine] in factorial
combinations. Shoots development was continued in MS plus 10microM BAP with and without
10 microM 2,4-D. Shoots were induced in media with 0.5 or 1.0 microM BAP. Shoots were
rooted in MS media with 2mg/l NAA and in 1mg/l each of NAA and IBA. CalliStarted to form
2-3 days after inoculation and continued proliferating with continuous subculture every 10-14
days. These formed initially along margins and veins of explants. The highest frequency of callus
formation (100%) was in media with 10microM each of 2,4-D in BAP. On the other hand, shoot
elongation and development occurred 20-28 days after inoculation. Shoot buds were formed in
media with only microM BAP and in 10 microM each of 2,4-D in BAP and 2,4-D. More shoots
elongated in media with 0.5 or 1.0 microM BAP compared with 10microM BAP. Elongated
shoots rooted in MS Medium with only 2,g/l NAA and in 1mg/l each of NAA and IBA. This
study suggests viability of applying the indirect organogenesis method for propagation of this
particular forest species and conservation.

SYNTHESIS OF THE REVIEWED STUDIES
From what the researchers have gathered, it shows that diabetes is the common diseases
not only in the Philippines but also in other countries.
The study on hand is supported from the related literatures and studies that were gathered
by the researchers. The similarities of this study with the previous study are process, and their
differences are the raw material used and this study only focuses on lowering the blood glucose
level.
The researchers used the data gathered for the experimentation. The researchers had
finalized to assess the potential of Noni (Morindacitrifolia) leaves extract in lowering blood
glucose level of Swiss Albino mice (Musmusculus).










CHAPTER III
METHODOLOGY
This chapter presents the research instruments, the research methodology and the
statistical treatment of data.
RESEARCH PLAN:
A. Research Design
The research design used in the study is complete randomized design (CRD).
Subsequent treatment is applied to determine the cause of change and the experimental
subjects are studied only once.
B. Material and Methods
250 grams of Noni (Morindacitrifolia) leaves
2 containers for Noni (Morindacitrifolia) extract
10 pieces of Swiss Albino mice (Musmusculus)
5 pcs cage
10 pieces of disposable syringe
Feeds for mice
Chocolate and softdrinks
150 mL distilled water
Gloves and Cheesecloth
Accu-Chek Meter Kit and Strips


C. Research Methodology
I. Gathering of Noni (Morindacitrifolia) leaves
The researchers had collected 250 grams of Noni
(Morindacitrifolia) leaves near the Satellite, Pacita, San Pedro, Laguna.
II. Acquisition of Swiss Albino mice (Musmusculus)
Ten Swiss Albino mice (Musmusculus) were obtained in Pacita.
The mice were divided into four groups regardless of sex.
GROUPINGS:
Control set-up 2 mice
Trial 1 (25% concentration) 2 mice
Trial 2 (50% concentration) 2 mice
Trial 3 (75% concentration) 2 mice
Trial 4 (100% concentration) 2 mice
III. Initial Blood Checking
The blood sample was collected using syringe for initial blood
checking. Then, the mice were being fasted for four hours.
IV. Feeding of Swiss Albino mice (Musmusculus)
The mice were fed with crushed chocolates and softdrink. This has
been conducted to make the mice positive to high level of blood glucose.

V. Preparation of Treatment
The Noni (Morindacitrifolia) leaves extract is acquired by soaking
the leaves into container overnight using cheesecloth.
VI. Application of Treatment
The mice were induced orally with Noni (Morindacitrifolia)
extract for the control set-up.
VII. Blood Glucose Level Checking (after fed)
Using the Accu-Chek Meter bought from Watsons, the blood
glucose was checked.
VIII. Tabulation of Results
The data were tabulated using the tables below.
Extract from Noni
(Morindacitrifolia)
Initial Blood Glucose
Level (mmol/L)
Blood Glucose Level
after fed (mmol/L)
Blood Glucose Level
after treatment
(mmol/L)
Control
25% concentration
50% concentration
75% concentration
100% concentration





XI. Statistical Tool Used
A non-parametric test was used in the study. The Wilcoxon Signed Ranked Test
was applied to test the hypotheses stated in the study. This was utilized instead of the t-
test because of the class size and the results from the Accu-Chek meter kit was not large
enough to calculate by parametric tests.
BIO-STATISTICAL TREATMENT
1. Arithmetic Mean
This is the most commonly used statistical method to measure the central
tendency. It is also called the computed average. This will be used to compute the
average blood glucose of the Swiss Albino mice (Musmusculus).
Formula used to get the mean for the ungrouped data.
X =
Where:
X = mean
= value of each item
number of items in the sample




2. Wilcoxon Signed Rank Test
Wilcoxon Signed Rank Test is a better test because it takes into account the
magnitude of the observed differences between paired values in the two samples. The
original signs of the differences are then given to their respective ranks.
Steps in doing the signed rank test:
a. State the null hypothesis.
b. State the alternative hypothesis.
c. Obtain the differences between paired values in the two samples/treatments.
d. Rank the absolute values of these differences, giving the smallest difference
rank 1 and assigning average ranks to tied differences.
e. Assign to each rank the sign of observed differences.
f. Get the sum of the positive ranks and the sum of the negative ranks.
g. Find the smaller of the absolute values of the two sums of the ranks and
denote it by T.
h. Test H
o
by comparing T with critical values of T for the Wilcoxon Signed
Rank Test. If T is less than or equal to T
critical
for the chosen level of
significance, a, then the null hypothesis is rejected. Otherwise the null
hypothesis is rejected.




Acquisition of Swiss Albino mice
(Musmusculus)
D. Treatment/General Procedure
The flow chart methodology of the study

Acclimatization
Feeding with chocolate
Measurement of Glucose level (after fed)
Collection and Extraction of Noni
(Morindacitrifolia) leaves
Application of Extract to Swiss Albino mice
(Musmusculus) with varying concentrations
25%, 50%, 75%, and 100% as oral
suspension/solution
Measurement of Glycaemic response of Swiss
Albino mice (Musmusculus) to Noni
(Morindacitrifolia) leaves extract
Measurement of GlucoseLvl (Initial Value)