University Research Proposal: Beneficial Antioxidant Effect of I3C

This research proposal investigates the beneficial antioxidant effects of Indole-3-Carbinol (I3C), a compound found in cruciferous vegetables. The study aims to evaluate I3C's potential in reducing oxidative stress, preventing cancer, and improving overall health. The rationale emphasizes the importan...Read More

Running head: BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
Name of the Student:
Name of the University:
Authors note:

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BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
RESEARCH PROPOSAL
Rationale of the Study:
The anti-oxidants [1] are the substance, which can be formed naturally or can be man-
made. Properties of antioxidant can potentially reduce the oxidation which can then be used
to reduce the cellular or tissue damage [2] in the biological entity like cells and tissues. An
effective form of anti-oxidants are Vitamin C [3] and Vitamin E [4]. Vitamin E, α-
tocopherols [5] can be found in vegetables, fruits, some types of nuts and other food sources.
Therefore, in a diet vegetables and fruits are one of the major sources of anti-oxidants. The
presence of antioxidants in the diet could reduce the prevalence of cancer [6] and
cardiovascular diseases [7]. It is to be understood that there are certain kinds of oxidative
stress which can be either external or internal that could cause certain categories of oxidative
stress[8] in the body like cigarette smoking[9], exposure to certain kinds of radiation and
other kinds of stressors [10]. In the current biomedical research, the establishment of anti-
oxidant property producing effective results in maintaining health processes and reduces the
oxidation in the body. Could also possibly reduce the inflammation and tissue damage. The
compound Indole-3-carbinol [11], is product formed by the breakdown of glucocobrasscin
[12], a substance found mostly in the family of Brassicaceae or Cruciferae. There are certain
plants have phytochemicals that could reduce the oxidative stress, and there could be
possibility upcoming of therapeutic effect which could produce the side effects. Therefore,
this particular family of plants, Brassicaceae could produce effective anti-cancerous [13],
cytotoxic effects [14] and other health benefits that could be effective. This group has daily
intake of the as a daily dietary supplement. The generation or production of free radicals
could increase the cause of certain diseases like atherosclerosis [15], gastritis[16], and aging
as well as liver diseases[17]. This is due to the production of super oxide, hydroxyl radicals
and other reactive species that is produced during the aerobic metabolism in the body that

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could cause excessive oxidative damage [18] in the body. The oxidative stress could further
cause the imbalance in the oxygen species [19] and anti-oxidants that could metabolic
disorders in the system. Hence, it is beneficial if the understand the efficacy of the natural
antioxidants available, that could be helpful in eliminating as well as alleviating many
diseases. The compound Indole 3 Carbinol, is compound belonging to the cruciferous
compound that have been proven to be a potential source of cancer eliminating property.
Apart from understanding the beneficial effect of this compound Indole 3 Carbinol it is
important to understand the chemical establishment dedicated in this anti-oxidant or anti-
cancerous properties in the Brassica vegetables, and understand the protective factors when it
comes to the development of the anti–oxidant properties and applicability in human beings.
Therefore, it is important to consider to the chemical properties of Indole 3 Carbinol, which is
a dimeric structure[20] which has to be more efficiently tested and investigated on the people,
in which there has to be in vitro as well as in vivo values should be analysed and propagated
accordingly. The importance of bioactive compound could further help us to understand the
detailed mechanism of its anti-oxidant properties like detoxification[21] of the enzyme,
induction of apoptosis as well as cell cycle regulation of the physiological processes. The
organic compound Glucosinolates [22] are the reason the cruciferous vegetables have typical
pungent smell, furthermore, there are certain reasons due to which the glucosinoalates could
also be a natural protective substance, which could be used against the pesticides, therefore,
also acts as a natural pesticides. In general, the reactive oxygen species The anti-oxidant
property of Indole 3 carbinol could be an effective source of reducing the oxygen free
radicals that could lead the betterment in the metabolic process of the cells and therefore,
maintain help a better regulation of life. The presence of this compound is evidently present
in our daily life style diet, and hence, finding out the antioxidant properties against certain
metabolic disease, like carcinogenesis and tumour growth could influence the cellular process

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and maintain better quality of life. Therefore, this study has aimed in finding the clinical
applicability of the random control trial[23], in case of patients who need anti-oxidant
therapy. It is important to understand and analyse the cellular physiology[24] as well as the
pathophysiology of the antioxidant effect of the indole 3 carbinol, could be analysed by
conducting a thorough experiment that would require the proper methodology of the data
collection as well as the descriptive research[25]. By this way it could be held for period of
time, and that could be investigated by assessing the follow ups respectively. The entire
process would need time around six months out which 3 months would be required for the
data collection and follow up visits, however, there could be extension in terms of data
processing as well as analysing.
Reason for Investigation:
The plant materials present in the human diet could possibly reduce the development
of many other metabolic diseases like cancer, and other metabolic diseases[26]. The plant
products could cause certain decrease in the cancer risk in the body, therefore the recognition
of the diet could be acting as a primary and a very vital causative factor when it comes to the
everyday diet that we follow. Hence it is important to evaluate the phyto-constituents
available and understand the anti-oxidant activity [27]. The indole-3-carbinol product could
be useful in many biological problems and issues and furthermore, there are acid catalyst
action in the stomach could further produce the indole 3 carbinol product which is bioactive
[28]. Hence, in this research proposal, the aim of the study is to find the beneficial effect of
the anti-oxidants in indole 3 carbinol would be evaluated in terms of further lab research as
well as literature survey, qualitative data and to understand how it can be used in future
prospect and research. It is important to understand the efficacy and therapeutic
establishment of this family of plant and understand the beneficial anti-oxidant in plants.
Moreover, the beneficial effect of the indole-3-carbinol could give more positive health

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effects, and that’s why good analysis of the anti-oxidant property of the compound should be
reviewed from the available literature.
Statement of Problem: In the recent time, there is a growing concern about the diet and life
style affecting our body physiology and metabolism. The metabolism related disorders are
increasing day by day and therefore, it is to be understood that there is a huge importance of
this substance in our diet. Moreover, the biological availability[29] in the body, could be
analysed on the basis of that the indole 3 carbinol molecule could be generated by the
stomach from the conversion of glucobrassin in to indole 3 carbinol. This generated in bio
active indole 3 carbinol oligomers like diindolylmethane or DIM and 5, 11- Dihydroindolo-
[3, 2, b] carbazole or ICZ. There are certain hormone dependent cancers which could result
in carcinogenic attributes in the body however, it has be understood that from the prior
research it could be inferred that these compounds that is I3C [30] and DIM [31] are
responsible for the bio transformation of the compounds that could be responsible for the
removal certain biological compounds which includes certain kinds of toxins [32],
steroids[33] and carcinogens[34] from the body. Hence, in that way it could also eliminate
the hormone dependent anomalies that also reduce the risk of hormone dependent cancers.
The optimisable amount of the dose is an important factor, and hence, it is important to
understand the fact the effect of anti-oxidant could further eliminate or neutralise the effect of
the reactive oxygen species that could cause cellular damage in the cells. Therefore, right
dose and right administration is very important to analyse the anti-oxidant properties of
Indole 3 carbinol. There is a strong relation of the anti-oxidant effect on the cellular reactions
and DNA regulation any alteration to the DNA could induce carcinogenesis and other
oxidative stress related issues. The evaluation of the anti-oxidant activities and proper cellular
regulation could help in proper administration as well as proper dosage of the 13C
supplement on the people. However, there are not enough research done on the established

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anti-oxidant properties in terms of other metabolic diseases and proper dosage or amount o of
the 13C supplement in our diet or as medicament could not be evaluated yet. The
incorporation of anti-oxidant properties in creating possible regulation of the metabolism as
well as, however, According to Marengo and other authors [35], there has to be an optimal
dosage when it comes to the effect of the antioxidants in case of cancer, it could be
understood that there could be increase of reactive oxygen species if there could be the
cytotoxic and also diminish the beneficial effect of the antioxidants in the cells. Therefore, it
could be inferred that there should be optimal dosage of anti-oxidants when it comes to the
implementation of antioxidants properties in metabolic disorders like cancer and diabetes
[36]. There are many effects of antioxidants in cancer treatments, some are also in
combination form, however, therefore, the anti-oxidant effects of Indole 3 carbinol with the
optimal dosing has not yet been identified. The optimal dosing of Indole 3 carbinol as an
antioxidant effect in terms of cancer and other metabolic diseases, is not yet recovered.
Hence, in this research proposal the optimal dosing of the antioxidant property of the indole
3 carbinol has to be analysed by the random control study, where the administration of the
dose of indole 3 carbinol and its effect on the tumour growth has to be studied and evaluated.
Background of the Study:
There are numerous benefits of antioxidants in the maintaining a proper homeostasis
and balancing of the reactive oxygen species and antioxidants. Therefore, it has to be
understood that anti-oxidants play a crucial role in inhibiting the oxidation and initiate the
normal processing of cells as well as reduce the oxidative stress in the body. By this it could
be inferred that the use of indole 3 carbinol as an effective antioxidant could be efficient and
effective in reducing the carcinogenesis and tumour growth in the body. The dietary
supplement of the indole 3 carbinol includes Brassicaceae like cabbage, broccoli mustards
and crucifers, in general it is known as the cabbage family. This family is also known as the

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mustard family. The anti-oxidant effect of this plant family could be found in promoting anti-
cancer properties as well as could be termed as protective metabolite. The Brassicacease
plants have glucosinolates which are of special consideration that includes isothiocyanates
[37] which are formed after the degradation of those products. However, there are specific
scientific benefits observed in this botanical family and health promoting agents that could
lead to the better health outcomes[38] and better health benefits as well. Therefore, special
preference has been provided to this particular family of plant which can be usually observed
in our daily dietary lifestyle. The special anti-oxidant activity of this class of plants could
help also in prevention of ageing and ageing related diseases which also includes the related
cognitive and related disorders in aged people. Furthermore, these can be potential source of
many kinds of phytomedicines and could be effective against certain kinds of kidney
problems like hyperuricemia as well as gout. Along with antioxidant properties it could also
influence the central role dietary consumption and hence, could influence the intake of
nutrition and hence, could also influence the consumption of antioxidants. There are
observable beneficial antioxidant property of the in case of certain neuronal death which
could cause Alzheimer’s disease and also improve the cognitive property of the brain. The
antioxidants are emerging as the potential treatment in wide range of diseases, which includes
cancer as well. The chronic inflammation could also be assessed and controlled with the help
of the antioxidants. Therefore, there has to be given special attention to the antioxidants in
our daily dietary intake of and analysing the antioxidants properties which consist of natural
phytochemicals. The presence of the phytochemicals which can be medicinal in plants and
vegetables could be crucial. However, there are cruciferous vegetables that could associated
with the reduced cancer incidence especially in the gastrointestinal tract as well as in other
sites. The antioxidants also contain certain phenolic acids, polyphenols that are included in
producing antioxidants properties. The sulphur containing organic compounds could further

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BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
be associated with the compound glucosinolates as well as derived products. Hence, it is
important to understand the phytochemical efficacy in terms of the beneficial bioactive
substances that are found in Brassicaceae. There can be damage in terms of micromolecules
especially in cells if there can be increase of the reactive oxygen species, and could also be
prevalent in terms of carcinogenesis as well as the cardiovascular diseases. Furthermore, the
reactive oxygen species too play an important role in terms of the preventing disease, and
also helps in the assisting the immunological response in the body and maintaining a crucial
role in apoptosis. However, there has been constant questioning about the antioxidant
properties as well as the excess carcinogenicity that could increase. The supplement of the
antioxidants in the diet though could create and effective source of plant compound however,
there implementation of the antioxidant as drugs and therefore, could be used as a preventive
medicine as well. There are other active antioxidants and phytochemicals that could
constitute in prevailing the anti-allergic, anti-microbial, anti-inflammatory, antioxidant,
vasodilatory effect, cardio protective and anti-thrombotic activity. There is potential and
better nutritive value when it comes to the antioxidant capacity of Brassicacea which has
sulphur containing compounds. The cruciferous vegatables like broccoli, cabbage and
cauliflower, has been creating many beneficial effects that could influence cellular defence
detoxifying as well as influence the modification of epigenetic mechanism. Therefore, there
could be potential preventive effect that could be effecting the molecular targets as well as
has epigenetic alterations that could influence the phytochemical structure and evaluation of
the anti-oxidant as well as could be working as derivatives of inhibitors of carcinogenesis.
In our daily lives, the metabolic diseases are growing day by day, like obesity, cancer,
diabetes and many more, which creates abnormal chemical reactions and alters the normal
mechanism of the body. The reactive oxygen species and their cell signalling process imparts
an important role in terms of physiology as well as pathophysiology, Therefore, it is

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important for us to understand the imbalance created by the properties of free radicals and
antioxidants, in our daily diet. When there is an increase of the oxidative stress in the body
that could also lead to the cellular damage of the proteins, lipids as well as DNA that could
possibly lead to carcinogenesis and other metabolic ailments and disorders. Therefore, the
balance of oxidants and antioxidants is important for proper functioning and processing of
life. There are many observable antioxidant properties of the family of Brassicaceae, and that
could be effective and have a medicinal effect on the body. Due to the imbalance of the
oxidants and antioxidants that effects the acid base balance which could further effect the
mechanism of the biological homeostasis. Indole 3 carbinol, which is found in the as a
compound in the cruciferous vegetables that could be formed by the breakdown of the
glucobrassin.
Research Design
Research design refers to the design of the research conducting procedures. It is the
foundation of the research procedure or methodology [39]. The data collection, sampling
strategy and data analysis of any research are based on the research design. Before finding the
appropriate design for this research the research philosophy and approach should be
identified. The research philosophy of a can have three options to choose from namely
positivism philosophy, interpretivism philosophy and realism philosophy [40]. The
positivism philosophy allows the researchers to analyse numerical data by collecting data
from practical field through scientific procedure. According to purpose of this research, this
research needs an effective empirical research structure [42] and therefore the positivist
research philosophy will be chosen. For research approach, this research can follow either
inductive or deductive or abductive research approach. The inductive research approach
allows proposing a new framework, model or theory [41]. The deductive model [43]
however allows deducting the present method and their significance. In this research the

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effectiveness of Indole-3-Carbinol will be analysed. Therefore, the approach of this research
will be deductive.
Based on handing the objective of the research the research design is classified into three
categories namely, exploratory research, explanatory research and descriptive research. The
exploratory research allows exploring the unknown factors of a particular topic. The
explanatory designs helps to explain the underlying factors behind the key topic considering
their interaction and logical connection. The descriptive research design is a design to
describe the effectiveness of a method while examining its process to propose further
implication. The purpose of the research is to examine the effectiveness of Indole-3-Carbinol
as an anti-oxidant. This research will require examining the effectiveness through field based
data collection while proposing the implications of it as an effective anti-oxidant drug to
reduce the abnormal cell growth that can lead to cancer. Hence for this research, considering
the purpose and anticipated procedures, the research design will be descriptive research
design.
The research designs could be also segregated into different categories depending on the data
collection and analysis method. In medical and pharmaceutical study the most utilised
research designs are participant observation, focus group study, experimental study,
secondary or archival study, non-experimental study, case control study, cohort study and
longitudinal study. Hence, to measure the anti-oxidant property of Indole-3-Carbinol both
cell culture as well as human based experiment could be possible. To measure the
effectiveness and propose Indole-3-Carbinol the research will need to use human participants
due to lack of scope for laboratory procedure. This study will use a group of human
participants who will be provided with Indole-3-Carbinol capsules and placebo capsules to
measure the effectiveness of medicine. Hence, the research procedure will adopt the

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randomized placebo controlled parallel group trial to measure the statistically significance of
effectiveness of Indole-3-Carbinol on cell growth of tumour.
Randomised control trial is a very popular research design that allows to test the effectiveness
of any medical compound. In this process a group of participants are considered as
experimental group and they receive the experimental medicine, however the other group
known as control group are provided with either no medicine or another medicine or placebo
capsule. As mentioned earlier, the placebo capsule which will be used in this study is a blank
capsule without any medicinal compound in it. This research will also use the parallel
intervention design which will also allow two groups of participants to be treated
simultaneously with different intervention techniques. In this study the experimental group as
well as the control group will be provided capsules with supply of 90 days. The experimental
group will receive 100 to 200 mg per day dose of Indole-3-Carbinol. On the other hand the
control group will receive a placebo capsule containing some general nutritious compounds.
This method will allow the researcher to investigate whether the Indole-3-Carbinol has better
capability to act as antioxidant, which can normalise the free radicals or oxidants in cell and
the resultant defect in cell DNA. The defect in cell DNA disrupts the cell-division cycle and
cause ununiformed cell growth that is the fundamental reason of tumour. Work in the
selection of the participants and the data collection and analysis procedures will be discussed
in the method and procedure section.
Research method and procedure
The method of the research emphasises the step by step procedure of conducting a
particular research. The purpose of this method is to examine the differences between two
groups in accordance with the changes in their malicious tumour growth. As discussed earlier
this research is based on randomised control triad design that needs primary data collection.

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The primary data collection is the procedure of collecting real time data from field based data
collection. In this experiment the resource of the data is the diagnosis report of two groups of
participants. In this study, 70 participants with Cancerous tumours or malignant tumours will
be engaged in this study with their consent. Among the 70 participants 35 participants will be
chosen randomly as experimental group and 35 participants will be chosen as control group.
The randomize sampling is a sampling strategy that allows a research to chose the target
samples through probability selection. The term probability selection refers to the sampling
strategy where each potential candidate has equal chance of getting selected. In this research
ensuring the reliability and minimising the biasing error is essential. Therefore, to segregate
the participants into experimental and control group randomised sampling will be used.
The purpose of the research is to examine the effectiveness of Indole-3-Carbinol as an anti-
oxidant. This research will require examining the effectiveness through field based data
collection while proposing the implications of it as an effective anti-oxidant drug to reduce
the abnormal cell growth that can lead to cancer. Hence, before the intervention an MRI
diagnosis will be used to detect the volume of the tumour. After undergoing through the
assigned interventions that is either experimental or controlled, the participants will be tested
by MRI after 90 days. During the intervention of 90 days the participants will spend their
normal lifestyle at their home under their self-monitored medication course. During invention
period of 90 days the experimental group will receive 100 to 200 mg per day dose of Indole-
3-Carbinol. On the other hand, the control group will receive a placebo capsule containing
some general nutritious compounds. After the 90 days of intervention period the MRI test
will be conducted again to measure the volume of the malicious tumours of the participants.
During the intervention period the participants will be provided with two sets of capsules,
where experimental group will be provided with Indole-3-Carbinol capsule and the control
group will be provided with placebo capsule. Both the groups will be instructed and even

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provided with the regular medication guidance. The participants will spend their normal life
while taking the doses on regular basis. Both participant groups will be also instructed to
contact with the researchers after 90 days or approximately three months of initialising
intervention process. After 90 days, when the participant will contact with the researcher, the
MRI test will be again conducted on both participant groups to measure their resultant tumour
Volume.
The data analysis process will be based on qualitative analysis. The data analysis process
could be based on two types of analytical techniques namely qualitative analysis and
quantitative analysis. The qualitative analysis allows the research to analyse the non-
numerical data through rational reasoning and interpretation. However, quantitative analysis
is the process of analysing numerical data through various statistical and mathematical
procedures. This proposal proposed an experimental procedure through randomised control
trial. Hence in this situation, quantitative study will be the most viable data analysis strategy.
For this study, the increased volume of the malicious tumours of the participants will required
to be measured. The aim of this study is to analyse the difference between the volume
changes of the malicious tumours of experimental group and the control group. For this two
groups of participants two database tables will be developed, where each table will include
the participant identification number, their results of pre- MRI test and post MRI test. After
completing the experiment the paired T test statistical method will be used to compare the
difference between the mean difference of tumour size of both experiential and control group.
For the data analysis process the SPSS of version 2.0 data analysis software will be used. The
T test will also consider the 95% confidence interval among the results. The purpose of the
research is to examine the effectiveness of Indole-3-Carbinol as an anti-oxidant. This
research will require examining the effectiveness through field based data collection while
proposing the implications of it as an effective anti-oxidant drug to reduce the abnormal cell

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BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
growth that can lead to cancer. Therefore, to test the significance difference between two
groups the probability value of P value will be considered. If the P value is found to be lower
than 0.05, then it can be concluded that the difference between two groups are significant. On
the other hand, if the P value will be found higher than 0.05 that it should be concluded that
there is no significance difference between the two interventions.
Ethical considerations
It is crucial that ethical parameters should be decided from the start of the research.
Correct judgment while taking scientific decisions on behalf of the participants or while
taking a shared- decision with the participants to optimize the main aim of study – is very
important.
Most ethical issues are reported to rise from conflicting values between the researchers and
the participants or even between the participants as well. Consumption of research resources
by some of the participants to a greater extent as compared to other participants can lead to
ethical issues as well. Similarly, from the researcher’s point of execution, unequal
distribution of the research resources amongst the participants so that one gets more access to
the items and the other groups gets less – also leads to ethical conflicts. Any personal bias or
greater attention given to a particular participants can lead to ethical complications as well.
Dual relationships [44] are one of major ethical concerns that has to be considered and barred
by the researchers from the very beginning. Resolution of these dilemmas [45] has to be
correctly decided by the investigators of the research study. If the investigators find any
related clues of unethical practice or even a slight breaching of ethical boundaries, actions
must be taken. All these ethical dilemmas will be strictly considered and care will be taken
not to cross any of it. The randomized controlled trials will equally distribute the research

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BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
resources to all its participants and judge the results without any personal bias towards the
outcome of the research or towards a participant. At the very core, there are certain ethical
principles that should be followed such as (a) doing good - that is beneficence[46] (b) doing
no harm that is non-malfeasance [47]. In research practice, the following ethical principles
should be followed strictly (a) informed consent from the potential research participants must
be taken on strict basis (b) minimizing any risk of danger or harm to the research
participants; (c) protecting their confidentiality and anonymity [48] (d) avoiding deceptive
practices (e) giving research participants the free will or the right to be withdrawn from the
concerned research at any point of time.
A strict procedure must be followed from the inception of the experiment regarding
the consideration of ethical issues. A written consent will be taken from all the participants
which confirms their voluntary participation in the research experiment. The research setting
will be decided or chosen in such a manner that it extends comfort as in the personal and
professional sphere of the research participants. Precautions will be taken not to aggravate the
existing disease symptoms. The participants will be given the full freedom to participate in
the study, completely on their own will and leave the study whenever they feel so, at their
own discretion. Free will and voluntary effort is a vital ethical consideration of a research
study and will be given utmost important in this experimentation.
Protecting the confidentiality and anonymity of research participants is a crucially
important component of maintaining research ethics. Data collection method has to be
decided in such a way that it does not hurts (psychologically) the subjects participating in the
research. This generally occurs when the collected data is not taken care of confidentially
during analysis, storage or during the publication process (that is when dissertation
submission is to be marked). This, however, does not implies that types or the categories of
data collected from the research subjects are to be kept anonymous or confidential. It is

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BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
possible that an individual’s views and identity needs to disclosed at different stages of
research process (which is from data collection method to the publication of your research
dissertation). But, before doing so - permissions must be taken again from the participants
whether the confidential information should be disclosed or not. The subject must be
informed and explained why this disclosure is important. It can be done is an alternative way
that is by removing identifiers (e.g., names, vernacular terms, geographical cues and more) or
provide proxies when writing up. The proposed research will maintain the anonymity and
confidentiality of the research subjects so to eliminate any breaching of ethical barriers.
Personal information will be secured from the researcher’s side and facilitated as per the need
of the research.
There are certain deceptive practices which can compromise with research process and harm
the participants in an unethical way. There are certain questions which comes up in this
regard which are (a) why are the participants participating in this research (b) what does the
research needs from them if the participants are to be deceived? It makes the research
controversial and should always be avoided. Hence, the proposed research will strictly refrain
from any such practices so as not to breach the ethical barriers and perform the study with
complete dedication towards its participants.
Reliability and validity/ trustworthiness issues
Validity:-
Validity looks into the scientific fact whether the used instrument is completely in
conjunction with the variable data. Validity helps in understanding whether the instrument
applies for all content in regards to the variable or whether it successfully and rightfully
measures the construct. Validity can be of various types such as content validity, construct
validity, discriminant validity and face validity and the validity that fits to the research design

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BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
of this research study is construct validity. There is a subset of the mentioned content validity
which is face validity, where researchers gives their own opinion about whether the proposed
and used instrument assesses the intended aim of the study.
Construct validity is a validity tool that measures the extent of degree to which the
experimental test successfully assesses what the study claims to be analyzing. As mentioned
in the classical model of research study’s test validity, construct validity is considered as one
of the most cardinal tool in validating the research and others which are as important as this
one are criterion validity and content validity. Modernized validity theory which defines
construct validity as being one of the major concern in the field of validity research but it has
been used in a diversified quantitative researches. Construct validity gives the actual
appropriateness of the scientific inferences that has been made with regards to measurements
or observations. Constructs are often seen as abstractions which are deliberately devised by
researchers to formulate a latent research variable that is correlated with the scores on an
assessment scale.
There are certain barriers though, to the application of construct validity and should
be managed strictly while analyzing the test results. The barriers are 1. Hypothesis guessing:
If the research participant guesses or knows the about to be found out - end-result - then
participants behavior may alter given the pre conceptual bias. The behavior induced by the
knowledge of this paradoxical result which might or might not be the outcome – altered the
participant’s perception towards the process of the experiment and gave rise to bias
complications. Bias is common in experimental research design (whether it is intentional or
non-intentional). An example of this kind of bias is given in Stephen Jay Gould's book "The
Mismeasure of Man”. For example - recent immigrants from eastern parts of Europe to
United States of America who are generally unfamiliar with baseball sport could not provide
right answers to the sport specific questions and it was immediately inferred that these

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Eastern European people have a lower intelligence. But the asked questions did not assess
intelligence any. It only measured the difference in cultural knowledge between the US
citizens and the immigrants which is very normal, given the period of time one has spent in
the particular culture and place and has become accustomed to a specific culture. It is also
possible that many people who lives in United States might not know the rules of the sport as
compared to sport fans. Hence asking the right research question is of utmost importance.
Researcher expectations can be directly or indirectly communicated to the research
participants non-verbally which can be used to elicit a desired effect. Double-blind
experimental designs should be devised to address the above problem. Confounding the
variables is the root cause of the observed research design effects which may be again due to
research variables which has not been measured or considered.
The construct validity that will be used in this research study will be homogeneity and
theory evidence. In statistical analysis of quantitative research data, homogeneity arise in
analyzing the properties of several datasets. The data sets then relate to construct validity of
convenient assumption as if the statistical properties of a data set can be somehow correlated
to the other dataset. In meta-analysis though, the process that mixes the data within and from
different types of studies, homogeneity assesses the similarities or differences between the
used or concerned studies. Homogeneity is generally analyzed to different degrees of
complexities. However, the questions of homogeneity should apply and comply with every
parameter and facet of the statistical quantitative distributions, including the location
parameter. Intermediate-level research study might be able to shift from seeing the variability
from different perspectives and can be studied further to analyze the skewed shifts. Questions
of homogeneity also apply to statistical joint distributions.

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BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
The proposed study will also test its findings for test evidence. It wold attempt to
understand and find an evidence of the behaviour being similar to the hypothesized
theoretical propositions of chosen construct which is to be measured by the chosen
instrument.
Reliability:-
Homogeneity or internal consistency is the assessed data in form of split-half
reliability, item-to-total correlation, Cronbach's α and Kuder-Richardson coefficient.
Cronbach's α is a vastly used test in determination of the instrument’s internal consistency.
This test gives the correlations averages in each and every permutation and combination of
the derived split-halves. Instruments that has questions can certainly have higher than two
answers that can be used in determination of the results.
Stability is another widely used test–retest reliability tool and is parallel to reliability
testing. Test–retest reliability is examined and re-assessed when a research instrument is
applied to same group of participants or research subjects– more than one time under
identical circumstances. Therefore, this research study gives of the instrument’s reliability.
Types like parallel-form reliability (also known as alternate-form reliability) works like test–
retest reliability tool.
Equivalence is another reliability parameter used to examine and find out inter-rater
reliability. The equivalent test comprises of a qualitative process which helps in determining
the agreement level among two and more than two observers.
All of these reliability parameters will the used in the proposed research study to
determine the trustworthiness of the research findings.
Potential implications and recommendations

19
BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
Based on the findings of the study, potential implications regarding the reliability of
the pharmacological pathway used by the drug will be deciphered and drug action on
different tumor types will be found out as well. Although, the study will be conducted with a
group of medically delicate subjects – every step will be taken just to study the action of drug
without hindering the patient condition. The drug might prove very successful in certain type
of tumors which will be found out in the course of the research but it also implies it might not
prove that successful in all the subjects. This is where medical and ethical considerations
would be considered and taken care of, very strictly.
The clinical findings will be then asserted to give recommendations to the national
and global healthcare framework and how to intervene with the research findings, to establish
long term and short term goals in the treatment process of the patient. The findings will set to
further future researches before applying on a public scale.

20
BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
References
1. Xie, Liping, Haihua Feng, Sha Li, Guoliang Meng, Shangmin Liu, Xin Tang, Yan Ma et
al. "SIRT3 mediates the antioxidant effect of hydrogen sulfide in endothelial
cells." Antioxidants & redox signaling 24, no. 6 (2016): 329-343.
2. Reiter, R.J., Mayo, J.C., Tan, D.X., Sainz, R.M., Alatorre‐Jimenez, M. and Qin, L., 2016.
Melatonin as an antioxidant: under promises but over delivers. Journal of pineal
research, 61(3), pp.253-278.
3. Rodríguez-Roque, María Janeth, Begoña de Ancos, Concepción Sánchez-Moreno, M. Pilar
Cano, Pedro Elez-Martínez, and Olga Martín-Belloso. "Impact of food matrix and processing
on the in vitro bioaccessibility of vitamin C, phenolic compounds, and hydrophilic
antioxidant activity from fruit juice-based beverages." Journal of Functional Foods 14
(2015): 33-43.
4. Hoene, Miriam, Martin Irmler, Johannes Beckers, Martin Hrabě de Angelis, Hans-Ulrich
Häring, and Cora Weigert. "A Vitamin E-Enriched Antioxidant Diet Interferes with the Acute
Adaptation of the Liver to Physical Exercise in Mice." Nutrients10, no. 5 (2018): 547.
5. Ding, Zufeng, Tom GP Saldeen, Pankaj Mathur, and Jawahar L. Mehta. "Mixed
tocopherols are better than alpha-tocopherol as anti-oxidantsas good as statins." Current
Research: Cardiology 3, no. 4 (2016).
6. Calaf, Gloria M., Francisco Aguayo, Consolato M. Sergi, Angeles Juarranz, and Debasish
Roy. "Antioxidants and Cancer: Theories, Techniques, and Trials in Preventing
Cancer." Oxidative medicine and cellular longevity 2018 (2018).

21
BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
7. Siti, Hawa N., Y. Kamisah, and J. Kamsiah. "The role of oxidative stress, antioxidants and
vascular inflammation in cardiovascular disease (a review)." Vascular pharmacology 71
(2015): 40-56.
8. Furukawa, Shigetada, Takuya Fujita, Michio Shimabukuro, Masanori Iwaki, Yukio
Yamada, Yoshimitsu Nakajima, Osamu Nakayama, Makoto Makishima, Morihiro Matsuda,
and Iichiro Shimomura. "Increased oxidative stress in obesity and its impact on metabolic
syndrome." The Journal of clinical investigation 114, no. 12 (2017): 1752-1761.
9. Andersson, Bengt-Åke, Shariel Sayardoust, Sture Löfgren, Lars Erik Rutqvist, and
Nongnit Laytragoon-Lewin. "Cigarette smoking affects microRNAs and inflammatory
biomarkers in healthy individuals and an association to single nucleotide polymorphisms is
indicated." Biomarkers (2018): 1-6.
10. Noiri, Eisei, Akihide Nakao, Koji Uchida, Hirokazu Tsukahara, Minoru Ohno, Toshiro
Fujita, Sergey Brodsky, and Michael S. Goligorsky. "Oxidative and nitrosative stress in acute
renal ischemia." American Journal of Physiology-Renal Physiology(2018).
11. Fujioka, Naomi, Vincent Fritz, Pramod Upadhyaya, Fekadu Kassie, and Stephen S.
Hecht. "Research on cruciferous vegetables, indole‐3‐carbinol, and cancer prevention: A
tribute to Lee W. Wattenberg." Molecular nutrition & food research60, no. 6 (2016): 1228-
1238.
12. Galletti, Stefania, Manuela Bagatta, Ferdinando Branca, Sergio Argento, Gina Rosalinda
De Nicola, Stefano Cianchetta, Renato Iori, and Paolino Ninfali. "Isatis canescens is a rich
source of glucobrassicin and other health‐promoting compounds." Journal of the Science of
Food and Agriculture95, no. 1 (2015): 158-164.

22
BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
13. Zhang, Qi, Fang Zhang, Kiran Thakur, Jun Wang, Hao Wang, Fei Hu, Jian-Guo Zhang,
and Zhao-Jun Wei. "Molecular mechanism of anti-cancerous potential of Morin extracted
from mulberry in Hela cells." Food and Chemical Toxicology 112 (2018): 466-475.
14. Wonderlich, John, Gene Shearer, Alexandra Livingstone, Andrew Brooks, Mark J.
Soloski, and Matthew M. Presby. "Induction and measurement of cytotoxic T lymphocyte
activity." Current protocols in immunology 120, no. 1 (2018): 3-11.
15. Santos, Raul D., Samuel S. Gidding, Robert A. Hegele, Marina A. Cuchel, Philip J.
Barter, Gerald F. Watts, Seth J. Baum et al. "Defining severe familial hypercholesterolaemia
and the implications for clinical management: a consensus statement from the International
Atherosclerosis Society Severe Familial Hypercholesterolemia Panel." The lancet Diabetes &
endocrinology 4, no. 10 (2016): 850-861.
16. Sugano, Kentaro, Jan Tack, Ernst J. Kuipers, David Y. Graham, Emad M. El-Omar,
Soichiro Miura, Ken Haruma, Masahiro Asaka, Naomi Uemura, and Peter Malfertheiner.
"Kyoto global consensus report on Helicobacter pylori gastritis." Gut 64, no. 9 (2015): 1353-
1367.
17. Chalasani, Naga, Zobair Younossi, Joel E. Lavine, Michael Charlton, Kenneth Cusi,
Mary Rinella, Stephen A. Harrison, Elizabeth M. Brunt, and Arun J. Sanyal. "The diagnosis
and management of nonalcoholic fatty liver disease: practice guidance from the American
Association for the Study of Liver Diseases." Hepatology 67, no. 1 (2018): 328-357.
18. Kohanski, Michael A., Anuj Tharakan, Nyall R. London, Andrew P. Lane, and
Murugappan Ramanathan Jr. "Bactericidal antibiotics promote oxidative damage and
programmed cell death in sinonasal epithelial cells." In International forum of allergy &
rhinology, vol. 7, no. 4, pp. 359-364. 2017.

23
BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
19. Choudhury, Feroza K., Rosa M. Rivero, Eduardo Blumwald, and Ron Mittler. "Reactive
oxygen species, abiotic stress and stress combination." The Plant Journal 90, no. 5 (2017):
856-867.
20. Upadhyaya, Pramod, Adam T. Zarth, Naomi Fujioka, Vincent A. Fritz, and Stephen S.
Hecht. "Identification and analysis of a mercapturic acid conjugate of indole-3-methyl
isothiocyanate in the urine of humans who consumed cruciferous vegetables." Journal of
Chromatography B 1072 (2018): 341-346.
21. Liu, Yang, Jie Li, and Yunfeng Lu. "Enzyme therapeutics for systemic
detoxification." Advanced drug delivery reviews 90 (2015): 24-39.
22 Johnson, T. L., A. T. Dinkova-Kostova, and J. W. Fahey. "Glucosinolates from the
Brassica vegetables and their health effects." In Encyclopedia of Food and Health, pp. 248-
255. Elsevier Inc., 2015.
23. Honeybul, Stephen, David Anthony Morrison, Kwok M. Ho, Christopher RP Lind, and
Elizabeth Geelhoed. "A randomized controlled trial comparing autologous cranioplasty with
custom-made titanium cranioplasty." Journal of neurosurgery126, no. 1 (2017): 81-90.
24. Frazier, Hilaree N., Shaniya Maimaiti, Katie L. Anderson, Lawrence D. Brewer, John C.
Gant, Nada M. Porter, and Olivier Thibault. "Calcium's role as nuanced modulator of cellular
physiology in the brain." Biochemical and biophysical research communications 483, no. 4
(2017): 981-987.
25. Colorafi, Karen Jiggins, and Bronwynne Evans. "Qualitative descriptive methods in
health science research." HERD: Health Environments Research & Design Journal 9, no. 4
(2016): 16-25.

24
BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
26. Incalza, Maria Angela, Rossella D'Oria, Annalisa Natalicchio, Sebastio Perrini, Luigi
Laviola, and Francesco Giorgino. "Oxidative stress and reactive oxygen species in
endothelial dysfunction associated with cardiovascular and metabolic diseases." Vascular
pharmacology 100 (2018): 1-19.
27. Erboga, Mustafa, Mehmet Kanter, Cevat Aktas, Umit Sener, Zeynep Fidanol Erboga,
Yeliz Bozdemir Donmez, and Ahmet Gurel. "Thymoquinone ameliorates cadmium-induced
nephrotoxicity, apoptosis, and oxidative stress in rats is based on its anti-apoptotic and anti-
oxidant properties." Biological trace element research 170, no. 1 (2016): 165-172.
28. Gai, Meiyu, Maxim A. Kurochkin, Danyang Li, Boris N. Khlebtsov, Luo Dong, Nadja
Tarakina, Robin Poston, David J. Gould, Johannes Frueh, and Gleb B. Sukhorukov. "In-situ
NIR-laser mediated bioactive substance delivery to single cell for EGFP expression based on
biocompatible microchamber-arrays." Journal of controlled release 276 (2018): 84-92.
29. Na, Joo-Young, and Jong-Tae Park. "Postmortem RNA stability and availability of
microRNAs in postmortem body fluids in forensic medicine." ROMANIAN JOURNAL OF
LEGAL MEDICINE 26, no. 2 (2018): 167-172.
30. Kim, Jae Kwang, and Sang Un Park. "Current results on the biological and
pharmacological activities of Indole-3-carbinol." EXCLI journal 17 (2018): 181.
31. Liu, Man, Rumana Yasmeen, Naomi Fukagawa, Liangli Yu, Young Kim, and Thomas
Wang. "Dose-Dependent Responses of I3C and DIM on T-Cell Activation in the Human T
Lymphocyte Jurkat Cell Line." International journal of molecular sciences 18, no. 7 (2017):
1409.
32. Clark, Graeme C., Nicholas R. Casewell, Christopher T. Elliott, Alan L. Harvey, Andrew
G. Jamieson, Peter N. Strong, and Andrew D. Turner. "Friends or foes? Emerging impacts of
biological toxins." Trends in biochemical sciences (2019).

25
BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
33. Candido, Kenneth D., Oscar J. Perozo, and Nebojsa Nick Knezevic. "Pharmacology of
acetaminophen, nonsteroidal antiinflammatory drugs, and steroid medications: implications
for anesthesia or unique associated risks." Anesthesiology clinics 35, no. 2 (2017): e145-e162.
34. Pohanish, Richard P. Sittig's handbook of toxic and hazardous chemicals and
carcinogens. William Andrew, 2017.
35. Marengo, Barbara, Mariapaola Nitti, Anna Lisa Furfaro, Renata Colla, Chiara De Ciucis,
Umberto Maria Marinari, Maria Adelaide Pronzato, Nicola Traverso, and Cinzia
Domenicotti. "Redox homeostasis and cellular antioxidant systems: crucial players in cancer
growth and therapy." Oxidative medicine and cellular longevity 2016 (2016).
36. American Diabetes Association. "2. Classification and diagnosis of diabetes." Diabetes
care 40, no. Supplement 1 (2017): S11-S24.
37. Dufour, Virginie, Martin Stahl, and Christine Baysse. "The antibacterial properties of
isothiocyanates." Microbiology 161, no. 2 (2015): 229-243.
38. Phillips, David E., Carla AbouZahr, Alan D. Lopez, Lene Mikkelsen, Don De Savigny,
Rafael Lozano, John Wilmoth, and Philip W. Setel. "Are well functioning civil registration
and vital statistics systems associated with better health outcomes?." The Lancet 386, no.
10001 (2015): 1386-1394.
39. Bowling, Ann. Research methods in health: investigating health and health services.
McGraw-hill education (UK), 2014.
40. Bhaskar, Roy. Enlightened common sense: The philosophy of critical realism. Routledge,
2016.
41. Krueger, J. I., & Heck, P. R. (2017). The heuristic value of p in inductive statistical
inference. Frontiers in psychology, 8, 908.

26
BENEFICIAL ANTI-OXIDANT EFFECT OF INDOLE-3-CARBINOL
42. İnce, Tolga, Hakkı Akman, Dilek Çimrin, and Adem Aydın. "The role of melatonin and
cortisol circadian rhythms in the pathogenesis of infantile colic." World Journal of
Pediatrics 14, no. 4 (2018): 392-398.
43. Tajani, Francesco, and Pierluigi Morano. "An empirical-deductive model for the
assessment of the mortgage lending value of properties as securities for credit
exposures." Journal of European Real estate Research 11, no. 1 (2018): 44-70.
44. Ryder, Robert, and Jeri Hepworth. "AAMFT ethical code: “Dual relationships”." Journal
of Marital and Family Therapy 16, no. 2 (1990): 127-132.
45. Csikai, Ellen L. "Social workers' participation in the resolution of ethical dilemmas in
hospice care." Health & social work 29, no. 1 (2004): 67-76.
46. Bates, Reid. "A critical analysis of evaluation practice: the Kirkpatrick model and the
principle of beneficence." Evaluation and program planning 27, no. 3 (2004): 341-347.
47. Gillon, Raanan. "" Primum non nocere" and the principle of non-maleficence." British
medical journal (Clinical research ed.) 291, no. 6488 (1985): 130.
48. Wiles, Rose, Graham Crow, Sue Heath, and Vikki Charles. "The management of
confidentiality and anonymity in social research." International Journal of Social Research
Methodology 11, no. 5 (2008): 417-428.