RESEARCH PROJECTS
To determine the diagnostic performance characteristics of blood-based protein biomarkers for the diagnosis of acute stroke using high-throughput proteomics approach (PhD thesis)
Duration of the project: August 2016 to February 2022 (5.5 years)
Description of the project: The diagnosis of stroke is largely clinical but treatment differs according to the type of stroke which at present can be determined with confidence only by neuroimaging. Sometimes clinically it is difficult to distinguish ischemic stroke (IS) from hemorrhagic stroke (HS). Recent developments in high-throughput proteomics approach have shown potential to discover biomarkers for distinguishing the two stroke types. Using high-throughput proteomics approach such as Liquid Chromatography- Mass Spectrometry (LC-MS/MS), serum samples will be assayed for the presence of potential protein biomarker(s) which can distinguish acute IS cases from acute HS cases and stroke from controls (with matched risk factors for stroke). The differentially expressed proteins with significant potential would further be validated in a large sample of patients to test their diagnostic performance and clinical significance. The study would help in the determination of blood based protein biomarkers for diagnosing and differentiating IS from HS, acute stroke cases from risk factor matched controls and healthy controls using high throughput proteomics approach with further validation in a larger sample.
A prospective, multicentric, randomized, double-blind, parallel, phase-III clinical study to assess efficacy of PMZ-1620 along with standard treatment in patients of acute ischemic stroke
Position: Clinical Research Coordinator (CRC)
Duration worked: January 2020 to Septemer 2021 (1 year & 9 months)
Funding body: Pharmazz India Pvt. Ltd. (Subsidiary of Pharmazz Inc. USA)
Description of the project: The peptide Sovateltide (IRL-1620) is a highly selective ETB receptor agonist. There are hidden stem cells in the brain, which become active following injury to the brain. Intravenous administration of PMZ-1620 (sovateltide) augments the activity of neuronal progenitor cells in the brain to repair the damage by formation of new mature neurons and blood vessels. In addition, PMZ-1620 has anti-apoptotic activity and also increases cerebral blood flow when administered following ischemia. It was discovered that in rat model of ischemic stroke, sovateltide, significantly improved survival, reduced neurological and motor function deficit while effectively decreasing infarct volume, edema and oxidative stress. The convincing results of preclinical efficacy studies of Sovateltide in ischemic stroke and its safety affirmation from phase I and phase II clinical studies encouraged us to investigate its efficacy in human patients of ischemic stroke in a phase III clinical study.
A prospective, multicentric, randomized, double-blind, parallel, saline, controlled phase-II clinical study to compare the safety and efficacy of PMZ-1620 therapy along with standard supportive care in subjects of acute ischemic stroke
Position: Clinical Research Coordinator (CRC)
Duration worked: February 2018 to August 2019 (1 year & 6 months)
Funding body: Pharmazz India Pvt. Ltd. (Subsidiary of Pharmazz Inc. USA)
Description of the project: Sovateltide (IRL-1620, PMZ-1620), an endothelin-B receptor agonist, administered intravenously following acute cerebral ischemic stroke increases cerebral blood flow, has anti-apoptotic activity and produces neurovascular remodeling. Its safety and tolerability have been confirmed in healthy human volunteers. This trial aims to determine safety, tolerability and efficacy of sovateltide as an adjuvant to standard of care (SOC) in 40 acute cerebral ischemic stroke patients.
Outcome of the project: Sovateltide was safe, well tolerated, and resulted in quicker recovery and improved neurological outcome in acute cerebral ischemic stroke patients 90 days post-treatment.
Empirical antibiotics in acute stroke patients with fever and their outcome: an ambispective cohort study
Position: Co-Investigator
Duration Worked: September 2017 to August 2019 (1 year & 11 months)
Funding body: AIIMS Intramural grant
Description of the project: Stroke with fever is associated with a poor outcome. Intervention studies of fever in stroke patients have been associated with mixed outcomes. We aimed to determine the factors associated with fever in stroke patients and their outcome at six months. We conducted an ambispective cohort study in patients of stroke admitted between January 2016 to July 2019. The demographic and clinical features of all patients along with the putative factors associated with fever were recorded. The patients were followed up for their outcome in terms of modified Rankin Scale (mRS) score at six months.
Outcome of the project: We observed that presence of fever in stroke patients was an independent predictor of poor outcome in our study. This is in line with the hypothesis that fever may be an indicator of poor outcome in stroke at six months, irrespective of standard management measures.
Biomarkers to enhance the accuracy and precision of prediction of short-term and long-term outcome after spontaneous intracerebral hemorrhage: a prospective cohort study
Position: Junior Research Fellow (JRF)
Duration worked: November 2014 to October 2016 (1 year & 11 months)
Funding body: Department of Biotechnology, Government of India
Description of the project: Small studies have suggested that biomarkers may be independent predictors of re-bleeding and prognosis in patients with spontaneous intracerebral hemorrhage (ICH); but none of them are established. No study, large enough to have power to examine all clinical, neuroimaging and laboratory parameters, have been done. This multi-centric study was designed to fill this gap. Specifically, the study aimed to examine whether the biomarker can significantly add to the predictive accuracy of prognosis of ICH. In this project we examined whether any of the biomarkers CRP level, serum glial fibrillary acidic protein, troponin, change in leukocyte count, S100B and copeptin levels are independent predictors of the neurological outcome in patients with primary intracerebral haemorrhage, and whether any of them (singly or in combination) improve the predictive accuracy of clinically important outcomes.
Nano-encapsulation of urease enzyme in alginate biopolymers and study of its enzyme kinetics (Master's thesis)
Project duration: March 2014 to June 2014 (4 months)
Description of the project: A procedure for the preparation of alginate nanoparticles in the aqueous phase of water-in-oil (W/O) nanoemulsions was studied. This method allows the preparation of finely dispersed emulsions without a large input of mechanical energy. With alginate concentrations of 1–2 wt % in the aqueous phase, emulsions showed good stability and narrow distributions of droplets with radii < 100nm. Gelation of the alginate was induced by the addition of aqueous salt solution of various ions (Ca+2, Sr+2, Ba+2, Ni2+) to the emulsions under stirring, and particles formed were collected using a simple procedure based on extraction of the surfactant on addition of excess oil. The potential of “low-energy” method for preparation of biocompatible nanoparticles was further exploited for the encapsulation of sensitive biomacromolecules i.e. urease enzyme from jack beans. Urease from Jack beans was immobilized in alginate beads and various parameters were compared and optimized.
Outcome of the project: This research demonstrated the potential application of a new nano-encapsulation method employed to enhance the stability and bioavailability of bioactive molecules for biosensor application.
Yeast transformation- To study the upregulation or downregulation of DNA repair mechanism under CoCl2 (Hypoxia mimicking agent) (Summer training)
Project duration: May 2013 to June 2013 (2 months)
Description of the project: Under the stress conditions there are thousands of processes which are affected and thousands of responses are generated inside the human body. A number of processes are affected by the stress conditions and these processes may be very vital for a particular cell to function properly; it can be cell proliferation, cell division, transcription or translation. It may also have adverse effects on the major cell process i.e. protein folding and functioning, leading to mutations and oncogenesis and some of the major effects can even be seen physiologically. The area of concern taken here is hypoxia which is deficiency of oxygen due to decrease in the atrial pressure of oxygen at high altitude. Apart from the fact that hypoxia has several major effects on the cell processes, DNA double strand breakage can be considered a vital one that also occurs under this stress. DNA double strand breaks are repaired by primarily two major processes in human body i.e. by Homologous Recombination (HR) and Non-Homologous End Joining (NHEJ). NHEJ is responsible for V(D)J recombination and does not require any template strand, therefore considered as an efficient method of repair. But it also carries the risk of mutations. Now under this stress where there occurs double strand breaks, NHEJ repair mechanism acts, but whether this repair mechanism is inhibited i.e. suppressed, downregulated or upregulated, is the major concern of the experiment.
Outcome of the project: The above study has confirmed that NHEJ repair mechanism is inhibited/ down regulated in yeast under CoCl2 stress conditions with the help of plasmid fidelity method. This also confirms the decrease in the activity of 10 NHEJ proteins under stress conditions which are involved in the DNA repair mechanism.
Use of fluorescence microscope versus light microscope in direct diagnosis of fungal infections (Summer training)
Project duration: May 2012 to June 2012 (2 months)
Description of the project: In this study, 102 samples of various patients suspected with fungal infection were studied. The samples were treated with KOH and then with Calcofluor white stain. All the slides were examined first by light microscopy and then by fluorescent microscopy. When viewed with a fluorescent microscope, fungal elements stained with Calcofluor white stain were sharply delineated from surrounding tissue and were easily identified. Calcofluor white stained the fungal hyphae and it fluoresced under fluorescence microscope, permitting rapid examination of fungal infection. Then the results of the light microscope and fluorescent microscope were compared.
Outcome of the project: It was concluded that when both methods were used for fungal detection, the yield of positive result was higher. Although in most cases fluorescence was noted, but was not intense.