Periodic Reporting for period 1 - H. Pylori-Scopy (Biophotonic Spectroscopic tool for Helicobacter Pylori diagnosis - a pathway for low cost clinical robotic device)
Berichtszeitraum: 2019-09-02 bis 2021-09-01
Problem statement and importance for society: Gastric Cancer is the fifth most common malignancy in the world and third leading cause of death. It has affected a major part of the world population, more than 70% cases (677,000) has occurred in the developing countries or in low-and-middle-income countries (LMIC). According to the International Agency for Research on Cancer, 89% of all gastric cancer cases are attributed to infection from Helicobacter Pylori, morphologically a spiral-shaped bacteria which grows underneath the mucosal layer of the human stomach. It is a well recognized germ, which has coexisted within the human stomach for many thousands of years. Its global prevalence was estimated to 4.4 billion people in 2015, which is more than half of the world’s population.
H. pylori has a unique, acid gated membrane channel, which effectively controls the amount of alkali produced by the bacteria to combat gastric acid which allows it to survive in stomach. For maintaining prevalence, H. pylori produce an enzyme called urease, which converts urea (present in gastric secretion) into ammonia and CO2. The production of ammonia (alkaline buffer zone) around h. pylori neutralizes the acidity of the stomach, making it more hospitable to colonize and survive. In the presence of gastric acid, the membrane channel increases the amount of urea entering the bacterial cytoplasm by 300-fold, resulting into a significant production of ammonia for endemic acid neutralization. Due to this, sufficient ammonia is produced to neutralize the area between inner and outer layer of the bacterial cell membrane. In addition, the helical shape of h. pylori allows it to burrow into the sub-mucosa, which is less acidic than the inside space, or lumen, of the stomach. H. pylori attach to the cells that line the inner surface of the stomach. Such behavior subverts the innate immunity and then modulates the adaptive immune system. As a result, the immune system of stomach is silenced against digested microbes. This condition may lead to formation of gastric ulcer in stomach and duodenum, which in turn causes indigestion. Once the h. pylori population helps in establishing the ulcers, these ulcers penetrate deep into the endothelium and start damaging the surrounding tissue. This causes bleeding or perforation in the stomach surface. Early diagnosis of H. pylori is important for understanding the pathology of gastric cancer and starting early therapy. But the high cost, ambiguous diagnosis and poor patient engagement of existing diagnostic methods lead to late/advanced stage diagnosis of cancer. Patients spend $700-$1800 for a standard 2-week triple-drug therapy. Such high cost in the LMIC is a burden where 33 million individuals face catastrophic health expense and cancer take their toll at the cost of human life.
Objective: My vision is to demonstrate the “proof-of-concept” of a device, with three different techniques together for (A) Single stop, low cost, easily accessible, portable, unambiguous, instantaneous gastric cancer screening, which results in (B) Increased patient engagement with no waiting for medical reports and quick medication. This project aims to reduce the cost by performing early diagnosis with single stop, easily accessible, portable, unambiguous, instantaneous screening device, which results in increased patient engagement with no waiting for medical reports and quick medication. This is made possible through the combination of laser-based Raman/Fourier Transform Infrared, and Chitosan-Magnetic-Swob spectroscopy, which will offer an addendum tool for h-pylori diagnosis using standard endoscopy, suitable not only for in-theatre but also for portable/accessible use in the LMIC regions.
H. pylori has a unique, acid gated membrane channel, which effectively controls the amount of alkali produced by the bacteria to combat gastric acid which allows it to survive in stomach. For maintaining prevalence, H. pylori produce an enzyme called urease, which converts urea (present in gastric secretion) into ammonia and CO2. The production of ammonia (alkaline buffer zone) around h. pylori neutralizes the acidity of the stomach, making it more hospitable to colonize and survive. In the presence of gastric acid, the membrane channel increases the amount of urea entering the bacterial cytoplasm by 300-fold, resulting into a significant production of ammonia for endemic acid neutralization. Due to this, sufficient ammonia is produced to neutralize the area between inner and outer layer of the bacterial cell membrane. In addition, the helical shape of h. pylori allows it to burrow into the sub-mucosa, which is less acidic than the inside space, or lumen, of the stomach. H. pylori attach to the cells that line the inner surface of the stomach. Such behavior subverts the innate immunity and then modulates the adaptive immune system. As a result, the immune system of stomach is silenced against digested microbes. This condition may lead to formation of gastric ulcer in stomach and duodenum, which in turn causes indigestion. Once the h. pylori population helps in establishing the ulcers, these ulcers penetrate deep into the endothelium and start damaging the surrounding tissue. This causes bleeding or perforation in the stomach surface. Early diagnosis of H. pylori is important for understanding the pathology of gastric cancer and starting early therapy. But the high cost, ambiguous diagnosis and poor patient engagement of existing diagnostic methods lead to late/advanced stage diagnosis of cancer. Patients spend $700-$1800 for a standard 2-week triple-drug therapy. Such high cost in the LMIC is a burden where 33 million individuals face catastrophic health expense and cancer take their toll at the cost of human life.
Objective: My vision is to demonstrate the “proof-of-concept” of a device, with three different techniques together for (A) Single stop, low cost, easily accessible, portable, unambiguous, instantaneous gastric cancer screening, which results in (B) Increased patient engagement with no waiting for medical reports and quick medication. This project aims to reduce the cost by performing early diagnosis with single stop, easily accessible, portable, unambiguous, instantaneous screening device, which results in increased patient engagement with no waiting for medical reports and quick medication. This is made possible through the combination of laser-based Raman/Fourier Transform Infrared, and Chitosan-Magnetic-Swob spectroscopy, which will offer an addendum tool for h-pylori diagnosis using standard endoscopy, suitable not only for in-theatre but also for portable/accessible use in the LMIC regions.
Summary of Work:
1. The project began with Raman spectroscopy characterization of Jack bean urease (synthetic sourced from Sigma Aldrich).
2. H. pylori supernatant was obtained from Old Medical School (Leeds General Infirmary) and Raman spectroscopy was performed on the supernatant.
3. pH based (1 to 7) experimentation was performed with Jack bean urease through Raman and FTIR spectroscopy.
4. Jack bean urease was then analyzed for its magnetic property through temperature/magnetic field based characterization.
5.40 pairs of stomach cancer and normal stomach tissues were sectioned on the cryostat (St. James Hospital, Leeds) and later analyzed via Raman/FTIR spectroscopy. Additional experimentation were performed via confocal imaging and H&E staining.
6.For the purpose of endoscopic device development for H. pylori diagnosis, an origami inspired soft robotic actuator was prototyped for upper gastrointestinal endoscopic application.
Summary of Results:
1. Detection of urea and urease compounds by using spectroscopic techniques (Raman and ATR-FTIR) was demonstrated.
2. Limit of detection of urea and urease compounds at different pH (1, 2, 3, 4 and 7) was demonstrated using Raman spectroscopic techniques.
3. Detection of different proteins of the Helicobacter pylori supernatant was demonstrated using Raman spectroscopy.
4. An Origami Actuator design of soft robotic endoscopic tip.
1. The project began with Raman spectroscopy characterization of Jack bean urease (synthetic sourced from Sigma Aldrich).
2. H. pylori supernatant was obtained from Old Medical School (Leeds General Infirmary) and Raman spectroscopy was performed on the supernatant.
3. pH based (1 to 7) experimentation was performed with Jack bean urease through Raman and FTIR spectroscopy.
4. Jack bean urease was then analyzed for its magnetic property through temperature/magnetic field based characterization.
5.40 pairs of stomach cancer and normal stomach tissues were sectioned on the cryostat (St. James Hospital, Leeds) and later analyzed via Raman/FTIR spectroscopy. Additional experimentation were performed via confocal imaging and H&E staining.
6.For the purpose of endoscopic device development for H. pylori diagnosis, an origami inspired soft robotic actuator was prototyped for upper gastrointestinal endoscopic application.
Summary of Results:
1. Detection of urea and urease compounds by using spectroscopic techniques (Raman and ATR-FTIR) was demonstrated.
2. Limit of detection of urea and urease compounds at different pH (1, 2, 3, 4 and 7) was demonstrated using Raman spectroscopic techniques.
3. Detection of different proteins of the Helicobacter pylori supernatant was demonstrated using Raman spectroscopy.
4. An Origami Actuator design of soft robotic endoscopic tip.
The current status of the result of this project are:
1. Spectral identification sub-components of urea, urease and Helicobacter pylori.
2. Spectral analysis of paired (cancerous and non-cancerous) stomach cancer tissues.
3. Magnetic characterization of Jack bean urease.
4. Exploration of endoscopic development.
These results are a foundation for future work of developing miniature optical fiber-based endoscope through the successful dissemination of above studies.
Future work for extending this research would involve:
1. Exploration of miniaturization techniques for introducing optical fibres into low-cost-soft robotic endoscopic devices. The work in this domain is planned to conducted on the outcomes of H.Pylori-Scopy with the GHRG for low-cost surgical technologies in LMIC (NIHR funded) for understanding its level of acceptance.
2. Experimental validation (comparing results with current studies) of developed device by performing Raman and FTIR spectroscopy on JBU, HPU, stomach tissues respectively. These experimental studies would be evaluated on the basis of sensitivity, specificity, repeatability and time consumed to obtain the results.
3. Magnetic characterization of JBU has helped us to understand biomolecular magnetic behaviour of natural occurring proteins. These results could be explored to develop sensitive magnetic sensors for in-vivo detection of h. pylori.
4. Additional studies may involve exploring utilization of the developed device for other bacterial infections like Clostridium Difficile, H. Pylori, and E.coli.
Socio-economic impact:
The above work has allowed me to gain skills for holistic development of a endoscopic solution by bringing knowledge from basic science, biology and engineering for a medical problem which has affected more than half population of the world. It is relevant from improving reproductive potential of male/female’s living in LMIC who really need medical support at low cost. This will positively impact health of general population with
single stop, unambiguous, instantaneous gastric cancer screening, with increased patient engagement, no waiting for medical reports and quick medication.
1. Spectral identification sub-components of urea, urease and Helicobacter pylori.
2. Spectral analysis of paired (cancerous and non-cancerous) stomach cancer tissues.
3. Magnetic characterization of Jack bean urease.
4. Exploration of endoscopic development.
These results are a foundation for future work of developing miniature optical fiber-based endoscope through the successful dissemination of above studies.
Future work for extending this research would involve:
1. Exploration of miniaturization techniques for introducing optical fibres into low-cost-soft robotic endoscopic devices. The work in this domain is planned to conducted on the outcomes of H.Pylori-Scopy with the GHRG for low-cost surgical technologies in LMIC (NIHR funded) for understanding its level of acceptance.
2. Experimental validation (comparing results with current studies) of developed device by performing Raman and FTIR spectroscopy on JBU, HPU, stomach tissues respectively. These experimental studies would be evaluated on the basis of sensitivity, specificity, repeatability and time consumed to obtain the results.
3. Magnetic characterization of JBU has helped us to understand biomolecular magnetic behaviour of natural occurring proteins. These results could be explored to develop sensitive magnetic sensors for in-vivo detection of h. pylori.
4. Additional studies may involve exploring utilization of the developed device for other bacterial infections like Clostridium Difficile, H. Pylori, and E.coli.
Socio-economic impact:
The above work has allowed me to gain skills for holistic development of a endoscopic solution by bringing knowledge from basic science, biology and engineering for a medical problem which has affected more than half population of the world. It is relevant from improving reproductive potential of male/female’s living in LMIC who really need medical support at low cost. This will positively impact health of general population with
single stop, unambiguous, instantaneous gastric cancer screening, with increased patient engagement, no waiting for medical reports and quick medication.