CFANR | K.S. Hegde Medical Academy (KSHEMA), Mangaluru

CENTER FOR ADVANCED NEUROLOGICAL RESEARCH (CFANR)


Genesis:
This unique center devoted to clinical and basic  research in the field of Neurology came into existence in 2006. The initial funding for starting the lab was brought in by Dr Lekha Pandit and a memorandum of understanding was signed with Nitte education trust on 15.07.2005. Subsequent funding came from the generous support of the Nitte trust and Governmental funding agencies such as DRDO, DST ICMR& DBT and international agencies such as Multiple Sclerosis Society of Great Britain.

It was inaugurated by Padma Vibhushan Professor MS Valiathan on 17.05.2006, in the presence of Dr Shantakumari (former Medical Education Director, Kerala), Professor IddyaKarunasagar (FAO Consultant and National Professor) and Dr KR Shetty ( Emeritus Professor of Neurology and former Dean, KMC, Mangaluru). The chief focus of research at CFANR is in the study of demyelinating disorders of the central nervous system. Both clinical and basic research is being done in the areas of Multiple Sclerosis ,Neuromyelitisoptica and other atypical CNS demyelinating disorders .

CLINICAL RESEARCH
Mangalore Demyelinating Disease registry
The growing number of patients with demyelinating central nervous system disorders seen at the Department of Neurology at K S Hegde Hospital led to the formation of a patient registry exclusively for the enrollment, assessment, subsidized treatment and dedicated long term followup of patients with Multiple sclerosis, Neuromyelitisoptica and other atypical demyelinating central nervous system (CNS) disorders. The Mangalore demyelination disease Registry, run by Dr Lekha Pandit was started in 2007 with a purpose to establish a database of CNS demyelinating disorders. Patients within a radius of 250 km from Mangalore are referred for specialist consultation and treatment. This registry has more than 1500 patients on its rolls. Nearly 2/3rd are followed longterm with dedicated treatment protocols and subsidized medications. Data from this registry has led to many original contributions that help to understand autoimmune neurological disorders in the Indian context.

Laboratory research

  1. Development of inhouse cell based assay for the detection of NMO IgG antibody using Indirect Immunofluorescence technique for the sensitive diagnosis of NMO IgG positive Neuromyelitis optica spectrum disorders.
  2. Evaluation of the genetic susceptibility for Neuromyelitis optica spectrum disorders.
  3. Biomarker development for the detection of unknown Antibodies associated with seronegative NMOSD.
  4. Evaluation of the role of certain Herpes viruses Human Endogenous Retrovirus in Multiple Sclerosis pathogenesis.

Externally funded projects
Title Funding Agency Amount
Development of a cost effective cell based assay for the diagnosis of Neuromyelitisoptica spectrum disorders in India DBT-BIRAC
2019-2021
Rs 40,93,000
Role of gut microbiome in central nervous system autoimmune disorders - A comparative study of patients of Indian origin with white Americans. Fulbright fellowship program , Brigham & Women’s hospital, Harvard medical school (host university)
2018-2019
15,000 USD (Rs 10,35,000)
Understanding the role of certain Herpes viruses & Human Endogenous Retrovirus inMultiple Sclerosis pathogenesis Dept of Biotechnology, Govt of India
2014-2017
Rs 61,77,600
Environmental & genetic risk factors for MS inIndian population Dept of Science &Technology, Govt of India
2011-2014
Rs 49,72,000
Study of patterns and prevalence of demyelinating disorders in urban Mangaluru Indian council of medical research
2010-2013
Rs 34,45,674
A longitudinal study to determine the clinical and radiological predictors for ADEM and MS and to evaluate underlying genetic susceptibility. Indian council of Medical research
2007-2010
Rs12,50,000

Ongoing Intramural funded projects:
Title Funding Agency Amount
Expression of immune genes analyzed using a Nanostring array (nCounter, Gene expression code set, Human immunology kit) in NMOSD Neurology Research Fund (2018-2019) Rs 6.5 lakhs
Establishing a rodent mouse model for Neuromyelitis optica spectrum disorders- A pilot collaborative study with Nitte university centre for animal research(NUCARE) Neurology Research Fund (2018-2020) Rs 5.00 lakhs
Standardization of immuno histochemistry techniques using a rodent rat brain model for diagnosis of seronegative Neuro myelitis Optica Spectrum disorders – A pilot study(in collaboration with NUCARE) Neurology Research Fund (2018-2020) Rs 2.5 lakhs


International Collaborative Projects:
Serial Number Title Collaborators
1 An international retrospective multi-center study of retinal optical coherence tomography in patients with neuromyelitis optica spectrum disorders
(2017-2019)
The Collaborative Retrospective Study on retinal OCT in Neuromyelitis Optica (CROCTINO) / Parallel Study on MRI in Neuromyelitis Optica (PAMRINO) project-

The Data Coordination Center (DCC) of the University of Utah,USA, Charité – Universitäts medizin Berlin-Germany, Guthy-Jackson Charitable Foundation, USA.
2 Comparative study of severity of Neuromyelitis Optica Spectrum Disorder world wide (2017-2019) Dr Jacqueline Palace Nuffield Department of Clinical Neurosciences, Oxford University.
Dr Ho Jin Kim National Cancer Center Korea, (Goyang-si South Korea)
3 Optic neuritis classification study 2018- An international collaborative study Lead collaborator- Dr Axel Petzold, UCL Institute of Neurology, Queen square, London,United Kingdom
4 An International survey on Treatment of MOG-IgG associated demyelination (2018-2019) Lead collaborator- Dr Anu Jacob, The Walton Centre NHS Foundation Trust Liverpool, United Kingdom


Facilities:
The research lab is equipped to do conventional and quantitative PCR , SNP genotyping and gene expression studies. In house facility for Western blotting, hybridization, indirect immunofluorescence testing, Oligoclonal band detection, Para protein detection and ELISA. We maintain cell lines for cell culture work and for novel biomarker development. The flow cytometer is being used for the stem cell enumeration ,monitoring of therapy after use of monoclonal antibodies ( eg. Rituximab) and also for the detection of other cell surface markers.



SI NO Instrument details Application Image
1 Flow cytometry
Model: BD FACS verse
Source of funding: Leela Narayan Shetty endowment fund
Used for cell surface marker detection, stem cell enumeration and monitoring of therapy after the use of monoclonal antibodies (Rituximab).
2 Inverted Fluorescent Microscope
Model:Eclipse Ti-S- Nikon
Source of funding:Neurology Research Fund
Used for Indirect immunofluorescence diagnostic tests and cell based assay development work
3 Class 2 biological safety cabinet
Model: 1300 series A2 -Thermoscientific
Source of funding: Leela Narayan Shetty endowment fund
Used for cell culture work
4 CO2 Incubator
Model: 371 Thermo Scientific Forma Air jacketed CO2 incubator
Source of funding:Leela Narayan Shetty endowment fund
Used for cell culture work
5 Cell counter
Model: Countess II FL- Invitrogen Bioservices
Source of funding:Leela Narayan Shetty endowment fund
Used for counting cells and to check cell viability
6 ELISA reader and washer
Model:iMark Microplate Absorbance Reader 168-1135 Bio-Rad
Source of Funding: Neurology Research fund
Used for diagnostic and research ELISA tests
7 MiliQ water purification unit
Model: MilliQ Direct 8S
Source of funding:Leela Narayan Shetty endowment fund
Used for obtaining ultra-pure water for culture and molecular biology work.
8 Real time PCR
Model: Applied Bio systems 7500 Real Time PCR system

Source of funding: DST project fund
Used for quantitative PCR, SNP genotyping and gene expression studies.
9 Thermal cycler
Model: S1000 Thermal Cycler with Dual 48/48 Fast Reaction Module

Source of funding: Neurology Research fund
Used for conventional PCR
10
Gel Doc unit Model: VilberLourmat Gel Documentation System

Source of funding: Neurology Research Fund
Used for imaging and documentation of nucleic acid and protein suspended in agarose and polyacrylamide gels.
11 Protean IEF system
Model:PROTEAN® i12TM IEF System and SDS PAGE unit,BioRad

Source of Funding: Leela Narayan Shetty endowment fund
Used for high throughput 2 dimensional protein separation
12 Hydrasys 2
Model: HYDRASYS 2 Iso Focusing - Automated Agarose gel Electrophoresis System.(Cat #:1203) Sebia, France
Source of funding: DBT project fund
Used for Oligoclonal Band and Para protein detection
13 Nanodrop-spectrophotometer
Model: Nanodrop 2000-spectrophotometer- Thermoscientific

Source of funding:Neurology research fund
Used for nucleic acid and protein quantification
14 Hybridisation oven
Model: Hybaid shake n Stack hybridisation oven- Thermofisherscientific

Source of funding: DST project fund
The hybridization oven is used for DNA, RNA or protein identification.
15 Incubators- 2 nos
Model: 1. Kemi-KIA1
Source of funding: DRDO project fund and clinical trial fund
Incubators provide a controlled, contaminant-free environment such as temperature and humidity for the growth and storage of bacterial cultures.
16 Laminar air flow unit-2 nos
Model: Laminar air flow-HLF3,HLF4- Labline industries
Source of funding: Neurology Research fund
Used to prevent contamination of biological samples or any particle sensitive materials. Air is drawn through a HEPA filter and blown towards the user.
17 Water bath-2nos
Model: 1.Serological water bath Equitron 2.Serological water bath-Rotek
Source of funding: DRDO project fund and Neurology research fund
It is used to incubate samples in water at a constant temperature over a long period of time
18 Centrifuge
Model:Thermo Scientific™ Sorvall™ ST 8R small benchtop centrifuge series
Source of Funding: Leela Narayan Shetty endowment fund
This centrifuge is used to separate serum, PBMC and plasma from blood.
19 Remi-Revolutionary Laboratory Centrifuge
Model : R-8C
Source of funding:Neurology Research Fund
This centrifuge is used in cell culture work
20 Remi Research Compufuge
Model : PR-23
Source of funding: DST Project Fund
This centrifuge is a used to separate blood products like serum , PBMC and plasma.
21 Remi Refrigerated centrifuge
Model : CPR-24
Source of funding: DST Project Fund
This centrifuge is used to separate DNA from blood.
22 Mini spin plus centrifuge
Model : Eppendorf 5453
Source of funding: DRDO Project Fund
This centrifuge is used to separate serum, plasma and DNA from blood.
23 Refrigerated Centrifuge
Model : Eppendorf 5702R
Source of funding: Clinical Trial
This centrifuge is used to separate serum, plasma and DNA from blood.
24 NBS Ultra Low temperature Deep freezer (-80°)
Model : U570-Eppendorf
Source of funding: Neurology Research Fund
This deep freezer is used for storage of samples
25 Thermo Scientific Forma Upright single door freezer-2 Numbers
Model : 906-ULTS
Source of funding: Leela Narayan Shetty endowment fund
1. This deep freezer is used for storage of barcoded DNA and serum samples
2.Used for storage of molecular biology reagents
26 Horizontal -20º Deep Freezer
Model : HF320
Source of funding: DRDO Project Fund
This deep freezer is used for storage of samples
27 Vertical -20º Scientific Deep Freezer - 2Numbers 1. Model : MDF-U5312-PE (Panasonic) YEN
2.Model : BFS 345S-SINGLE DOOR (Vestfrost)
Source of funding: Leela Narayan Shetty endowment fund and Clinical trial
This deep freezer is used for storage of cell culture media and molecular biology reagents
28 Refrigerator-2 Numbers
a.)Model :Ice Magic F23 Classic plus
b.) LG Refrigerator - GL-368YSQ4
Source of funding:Neurology Research Fund and clinical trial
Used for storing Reagents, media and diagnostic test kits.
29 LN2 container- 2 nos
Model : Thermo Scientific BioCane 47 Canister and Cane Systems
Source of funding: Leela Narayan Shetty endowment fund and Neurology Research Fund
Used for storing cells, tissues and other biological samples.
30 Pipettes:
a.)Serological pipette and Calibrated Micro Pipettes-6 sets(10µl, 20µl, 100µl & 1000µl)- Eppendorf
b.) Calibrated Multi Channel(10µl)- Nichipet-7000
c.)E4 XLS Electronic Multichannel Pipette- Rainin
Source of funding: Neurology Research fund
Used for Molecular biology and serological work.
31 Digital PH meter
Model: Hanna
Source of funding: Neurology Research fund
Used for reagent preparations
32 Autoclave Unit (Vertical)- 2
Model:Labline and Remi(India)
Source of funding: Neurology Research fund
Used for sterilizing pipette tips ,glass wares, media and other reagents