Research and Publication

Phase 1- At the Spectrophotometric Lab at the IISc.
Indian Institute of Science (IISc) is a public institute and university for research and higher education in science and engineering, Bangalore, Karnataka. It has been ranked 1st in the university and overall category for the last three consecutive years and has been considered a premier scientific research institute in India.

Research Title

My research is titled “QuitPuff: A Simple Method Using Salivary Lipid Peroxidative Changes to Assess the Risk of Oral Pre-cancerous Lesions and Oral Squamous Cell Carcinoma in Chronic Smokers 

Research Description

QuitPuff is a Point-of-Care salivary diagnostic test for early risk detection of oral cancer. The test is specially meant for chronic smokers who are at high risk of developing oral cancer.

It is a simple, quick, do-it-yourself test, requiring no special skills. Each test costs only Rs. 38 (approx. 50 cents) and has an accuracy of 96%. It can detect the risk early, at a stage when behavioural changes can be made and the risk can be reversed.

The Idea and Science behind Quitpuff


QuitPuff was tested on 500 subjects from Bangalore Medical College and Research Institute, Victoria Hospital, ESIS Hospital and Dr. Health Clinic. The subjects included non-smokers, smokers and smokers with oral pre-cancerous conditions and oral cancer. Informed consent was obtained from every subject.

The study was approved by the Institutional Ethics Committee. The sample analysis and experiments were conducted at the Indian Institute of Science, Bengaluru.

Research Motivation

I am a part of a clowning group and we believe “laughter is the best medicine”. So, we paint our faces, dress up as clowns, carry props and visit hospitals to cheer up the patients, make them smile and laugh.

On one such clowning visit at St. John’s hospital, I met a patient who was about 30 years old. He had a distorted face with almost no jaw on the right side. He could barely smile or open his mouth. When we tried to cheer him up, he cried instead. He was diagnosed with Oral Squamous Cell Carcinoma due to chronic smoking and had recently been operated upon.

This incident shook me up and I started googling about oral cancer. I read about the stages of oral cancer and the statistics. I read how detection of oral cancer at stage I carries a prognosis of 80% survival, while the same lesion at stage III carries a 20% survival. I looked for diagnostic tests that could detect the risk of smokers developing cancer at an early stage, but there were none. I only came across biopsies, CAT scans and MRIs that detect whether a lesion is cancerous or benign. But I found nothing that could detect the risk in the very early stages, much before the cancer lesion formed.

I kept googling and reading articles and journals and this was the only thing I would do after school. Most of what I read went over my head. It was only after reading them over and over again, did a few things make sense to me.

I read about the pathogenesis of cancer and about lipid peroxidation and how it can act as an early marker in certain cancers. Then I read about the various biomarkers of lipid peroxidation and came across Malondialdehyde and TBARS Assay and that got me working on devising a method which would detect the risk of a smoker or a tobacco chewer developing oral cancer. That is how my study started.

Globally, the 5-year mortality rate of oral cancer is approximately 50% and has not improved despite advances in diagnostic techniques and improvements in treatment modalities. The high mortality rate in oral cancer is attributed to late diagnosis, which is either due to lack of knowledge or access to medical care. Most patients seek help only in later stages when symptoms like pain, ulceration, or a neck mass appear.

Thus, there is a need for improvement in early risk detection of oral carcinomas, because in the initial and pre-cancerous stages, treatment is more effective and morbidity is minimal.

The need for early diagnosis of Oral Cancer and how my research aimed to solve it

The incidence of oral cancer is the highest in India, accounting for almost one-third cases found in the world. Smoking is found to be one of the leading causes of oral cancer. The international agency for research on cancer has predicted that India’s incidence of cancer will increase from 1 million in 2012 to more 1.7 million in 2035.

The high mortality rate is attributed mainly to late diagnosis as most patients seek help only in later stages when symptoms like pain, ulceration, or a neck mass appear.

Public health centers and private hospitals have recognized oral cancer as a grave problem and efforts towards early detection and prevention can help reduce this burden.

There is ample awareness that cigarette smoking can lead to cancer. For years, cigarette packets have been portraying images of cancer lesions, but this has not discouraged people from smoking. The number of smokers in India has risen from 79 million in 1998 to 108 million in 2015.

My research has attempted to address the need for early detection of oral cancer. I have devised a test that helps to identify the risk in a more specific and individualistic manner, in tangible terms, and at an early stage, much before any visible symptoms like oral lesion or neck mass have formed, at a stage when there could be a better chance of recovery. Using QuitPuff, any smoker can self-check his or her own risk and choose to make behavioural changes at the right time.

Research Publication

1)The Harvard GSAS Journal of Emerging Investigators

Nikhiya Shamsher and P. Chaithanya. “QuitPuff: A Simple Method Using Salivary Lipid Peroxidative Changes to Assess the Risk of Oral Pre-cancerous Lesions and Oral Squamous Cell Carcinoma in Chronic SmokersJournal of Emerging Investigators, vol 2, 2019.

2)The Columbia Junior Science Journal

Nikhiya Shamsher. “QuitPuff: A Simple, Home-based, Salivary Diagnostic Test to Assess Risk of Oral Pre-cancer and Cancer in Chronic Smokers” Columbia Junior Science Journal, 2019 (Page 25).

 

Reviewers Comments

    • An editor-in-chief has reviewed your manuscript and commends your efforts to start investigating an interesting scientific question! This is a very impressive piece of work coming out of a high school that exceeds the expectations of JEI. The editors and reviewers believe your study clearly articulates a problem and presents a creative approach to tackling said problem.
      The Harvard GSAS JEI Editorial Staff.
    • This paper does a great job at introducing a problem, the rational behind the proposed solution, and the results of applying the proposed solution. The utility of the results is clearly presented. Overall, I think that the work done in this manuscript is of high quality and worthy of publication in JEI”,
      By scientific reviewer who is a PhD Student Candidate, at Harvard University who researches functional antibody responses to influenza infection and vaccination, with the goal of finding ways to improve the flu vaccine.
    • This is a really impressive work and the framework would actually make a decent submission to a professional journal. As-is, it would be a standout submission to JEI.
      By Scientific reviewer who is PhD Student Candidate, at MIT who conducts molecular dynamics simulations of diffusion in nanocomposite membranes.
  • What a fascinating and clinically relevant project!”
    Scientific reviewer of Columbia Junior Science Journal.

Research Awards

Third Grand Award Winner at Intel ISEF 2019

As a Finalist from India I participated at the Intel-ISEF Fair at Phoenix, Arizona in May 2019 and won the Third Grand Award in the category of Biochemistry. I also earned an Honourable mention by the National Institute on Drug Abuse and National Institute of Health, USA. The competition had 1842 finalists from all over the world.

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Gandhian Young Technological Innovation Awards 2019

Received a research grant by the Government of India through the Gandhian Young Technological Innovation Awards 2019. The award was presented to me by the Honourable Union Minister of Health and Family Welfare, Dr. Harsh Vardhan in the presence of Vice President of India Shri M. Venkaiah Naidu at a ceremony held at Vigyan Bhavan, New Delhi on July 2019.

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Grand Award Winner at the Initiative for Research & Innovation in STEM (IRIS) National Fair 2018

Held in December 2018 at Manekshaw Centre, New Delhi – My paper was chosen from over 1500 entries as the best in the category of Biochemistry. As the Grand Award Winner, I was selected as a Finalist to represent India at the Intel-ISEF fair in 2019.

Gold Medal Winner at the National Indian Science and Engineering Fair (INSEF) 2019,

run by the Science Society of India, held in January 2019 at Shree K.G Dholakiya School, Rajkot.

Gold Medal Winner at the Regional Indian Science and Engineering Fair (INSEF) 2018,

run by the Science Society of India, held in November 2018 at Vagdevi Vilas Institution, Bangalore.

Finalist at the Pradeep Thevannoor Innovation Awards (PPTIA) 2018

held in November 2018 at the SCMS School of Engineering and Technology, Ernakulam, Kerala.

Phase 2- Design and the working of the QuitPuff Prototype. How does the test detect the early risk of oral pre-cancer and oral cancer?

Principle of the test

The test is based on color change produced upon heating saliva with the QuitPuff reagent. The intensity of color produced is directly proportional to the concentration of Malondialdehyde in the saliva sample and is indicative of the degree of lipid peroxidation and the risk of a smoker towards developing oral pre-cancer and cancer.

Working of the QuitPuff Salivary Lipid Peroxidation Kit

What does the QuitPuff test do?

The test checks the degree of lipid peroxidation in your saliva.

What is lipid peroxidation?

Lipid peroxidation is caused when free radicals damage the cells in your body.

Why should I check my salivary lipid peroxidation level? What does it indicate?

When you smoke, you generate free radicals. The free radicals attack the cells inside your oral cavity, causing damage to the structure and function of cell membranes. This process is called lipid peroxidation. It releases harmful substances that cause DNA damage and promote the development of oral cancer. Thus if you have been a chronic smoker, by checking your salivary lipid peroxidation level, you are able to assess the early risk of developing oral pre-cancerous conditions or oral cancer. This allows you to quit smoking at the right time and take charge of your health.

If my salivary lipid peroxidation level is high, does it mean I have oral cancer?

QuitPuff is a preliminary test for flagging the early risk. If your salivary lipid peroxidation level is high, it would be best for you to seek medical help for confirmation of diagnosis.

Who should do this test?

Smokers and tobacco chewers are at high risk and should do this test at regular intervals.

Directions for use

Refer to the QuitPuff Manual 

Societal Impact of my research

On account of its simplicity, non-invasive nature, low-cost and easy accessibility, QuitPuff can have great potential as a point-of-care test for oral pre-cancer and oral cancer.

Early Detection could reduce both mortality and healthcare costs

QuitPuff is proposed as an early detection tool for smokers to predict their risk of developing oral pre-cancer and oral cancer. If such a tool could be provided free of cost with every cigarette pack, then more people would be willing to check their risk and make behavioral changes prior to the development of oral cancer. More people taking these tests would mean more people finding out their risks in the early stages. Early detection could enable better treatment outcome and improvement in the quality of life. It could reduce healthcare costs and the economic burden of treating oral cancer.

Easy accessibility, storage & transportation to rural and remote locations

The test kit requires no elaborate storage conditions and could be easily transported to remote locations and stored in small pan-beedi shops (tobacco selling shops & kiosks) in rural areas.

Affordable

At per sample cost of Rs. 38 (approximately 50 cents), it could offer an inexpensive and affordable option especially to the lower income populations, where such a test is often most needed.

Ideal mass screening tool for large population samples

The test could be useful as a mass screening tool not only for routine clinics, but also for rural areas and remote locations with limited laboratory facilities or minimally trained health workers.

Phase 3- Future Development

In Phase 3, I aim to simplify the test further by devising a paper strip method or a lateral flow assay. A smoker can simply spit on a paper strip and within few minutes the color change would show his or her risk status, just like a home based pregnancy test.

My other goal is to improve the specificity and accuracy of QuitPuff and make it more specific to consumers of different tobacco products like beedi, paan, smokeless tobacco, betel nut, gutka, zarda, mawa, kharra and khaini.